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PMID:11096 | The fate of ACTH released from rat anterior pituitary into the incubation medium in vitro: enzymatic degradation and acid activation. | The bioactivity of ACTH released from isolated rat anterior pituitary glands into the incubation medium was determined. After the pituitaries were removed, ACTH activity in the medium decreased exponentially during further incubation at 37degreesC. The loss of ACTH activity was temperature- and pH-dependent and inhibited both by protease inhibitor (trasylol) and by preheating. Crude tissue extracts from median eminence, cerebral cortex and liver similarly inhibited the loss of ACTH activity. These results indicate that ACTH released into the medium may be destroyed by proteolytic enzyme(s) from the rat anterior pituitary. ACTH activity in the incubation medium was increased promptly by acidification of the medium to pH 1.5-2.5 with HC1, and reduced to the initial level by NaOH reneutralization of the medium (pH 6.8-7.8). These phenomena were not observed after the incubation medium had been heated at 100degreesC for 5 min. | The fate of ACTH released from rat anterior pituitary into the incubation medium in vitro: enzymatic degradation and acid activation. The bioactivity of ACTH released from isolated rat anterior pituitary glands into the incubation medium was determined. After the pituitaries were removed, ACTH activity in the medium decreased exponentially during further incubation at 37degreesC. The loss of ACTH activity was temperature- and pH-dependent and inhibited both by protease inhibitor (trasylol) and by preheating. Crude tissue extracts from median eminence, cerebral cortex and liver similarly inhibited the loss of ACTH activity. These results indicate that ACTH released into the medium may be destroyed by proteolytic enzyme(s) from the rat anterior pituitary. ACTH activity in the incubation medium was increased promptly by acidification of the medium to pH 1.5-2.5 with HC1, and reduced to the initial level by NaOH reneutralization of the medium (pH 6.8-7.8). These phenomena were not observed after the incubation medium had been heated at 100degreesC for 5 min. | [
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PMID:11097 | Ferredoxin from a red alga, Porphyra umbilicalis. | A plant-algal type ferredoxin was isolated from the red alga, Porphyra umbilicalis. In its oxidised form the ferredoxin had absorption maxima at 277, (281), 323, 420 and 462 nm. Two atoms each of non-haem iron and labile sulphur were present per molecule protein. The midpoint potential of the protein was -400 mV and it effectively mediated electron transport in the NADP-photoreduction system of barley. The amino acid composition of Porphyra umbilicalis ferredoxin was determined as (Lys4, His2, Arg1, Asx10, Thr8, Ser7, Glx16-17, Pro3, Gly7, Ala8, Cys5, Val6, Met1, Ile5, Leu8, Tyr5, Phe2). The minimum molecular weight of approximately 11000 was confirmed by sedimentation-equilibrium studies in the analytical ultracentrifuge. Approaching half of the total amino acid sequence was determined by means of an automatic sequencer. | Ferredoxin from a red alga, Porphyra umbilicalis. A plant-algal type ferredoxin was isolated from the red alga, Porphyra umbilicalis. In its oxidised form the ferredoxin had absorption maxima at 277, (281), 323, 420 and 462 nm. Two atoms each of non-haem iron and labile sulphur were present per molecule protein. The midpoint potential of the protein was -400 mV and it effectively mediated electron transport in the NADP-photoreduction system of barley. The amino acid composition of Porphyra umbilicalis ferredoxin was determined as (Lys4, His2, Arg1, Asx10, Thr8, Ser7, Glx16-17, Pro3, Gly7, Ala8, Cys5, Val6, Met1, Ile5, Leu8, Tyr5, Phe2). The minimum molecular weight of approximately 11000 was confirmed by sedimentation-equilibrium studies in the analytical ultracentrifuge. Approaching half of the total amino acid sequence was determined by means of an automatic sequencer. | [
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|
PMID:11098 | The antigen-inexperienced thymic suppressor cells: a class of lymphocytes in the young chicken thymus that inhibits antibody production and cell-mediated immune responses. | Transfer of thymus cells from young chickens in combination with a light whole body irradiation (360 R) was found to suppress the rejection of skin grafts across strong histocompatibility (B) differences. On the average, the suppressed animals also showed decreased serum hemagglutinin titers against erythrocytes of the skin donor strain and a decreased graft-versus-host (GvH) reactivity against embryos of this strain. The thymic suppressor cells can be obtained from animals that have not experienced the antigen under test. However, after transfer and contact to the antigen (skin graft) they can lead to the formation of specific ("activated") suppressor cells and can mediate in the long run a specific inhibition of the response to this antigen. The suppressive activity is associated with a bursa-dependent cellular subpopulation in the thymus that is different from B lymphocytes, B precursor cells or GvH-reactive T cells. The bursa dependency of the thymic suppressor cell suggests that functionally different lineages of thymic and thymus-derived lymphocytes are derived from different sources of prethymic stem cells. The suppressor cells are predominantly found in the young chicken thymus and already detectable in the 16-day-old embryo, while poor suppressive activity is found in the adult thymus. The suppressive effect can be obtained with thymus cells from either syngeneic or allogeneic donors. Embryonic allogeneic donors provide suppressive cell preparations free of GvH reactivity. The possibility that the thymus suppressor cells mediate self tolerance and "neonatal tolerance" is discussed. | The antigen-inexperienced thymic suppressor cells: a class of lymphocytes in the young chicken thymus that inhibits antibody production and cell-mediated immune responses. Transfer of thymus cells from young chickens in combination with a light whole body irradiation (360 R) was found to suppress the rejection of skin grafts across strong histocompatibility (B) differences. On the average, the suppressed animals also showed decreased serum hemagglutinin titers against erythrocytes of the skin donor strain and a decreased graft-versus-host (GvH) reactivity against embryos of this strain. The thymic suppressor cells can be obtained from animals that have not experienced the antigen under test. However, after transfer and contact to the antigen (skin graft) they can lead to the formation of specific ("activated") suppressor cells and can mediate in the long run a specific inhibition of the response to this antigen. The suppressive activity is associated with a bursa-dependent cellular subpopulation in the thymus that is different from B lymphocytes, B precursor cells or GvH-reactive T cells. The bursa dependency of the thymic suppressor cell suggests that functionally different lineages of thymic and thymus-derived lymphocytes are derived from different sources of prethymic stem cells. The suppressor cells are predominantly found in the young chicken thymus and already detectable in the 16-day-old embryo, while poor suppressive activity is found in the adult thymus. The suppressive effect can be obtained with thymus cells from either syngeneic or allogeneic donors. Embryonic allogeneic donors provide suppressive cell preparations free of GvH reactivity. The possibility that the thymus suppressor cells mediate self tolerance and "neonatal tolerance" is discussed. | [
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|
PMID:11099 | Inhibition of T cell activity in vivo: a test model for quantitative evaluation. | A test model is presented which, in comparison with the conventional models of skin transplantation or graft-versus-host (GvH) reaction in mice, permits a more sensitive quantitative evaluation of T cell inhibition in vivo. Prospective donors (type AA) are immunized with prospective recipient material (type AB); the resulting T cell reaction of A versus B is inhibited by consecutive treatment. Extent of inhibition can be evaluated after transfer of the pretreated AA material onto AB recipients by calculation of remaining GvH reactivity, if compared to adequate control tranfers. In this model the target animal for T cell reactivity (the AB recipient) remains untouched from immunosuppressive regimen. | Inhibition of T cell activity in vivo: a test model for quantitative evaluation. A test model is presented which, in comparison with the conventional models of skin transplantation or graft-versus-host (GvH) reaction in mice, permits a more sensitive quantitative evaluation of T cell inhibition in vivo. Prospective donors (type AA) are immunized with prospective recipient material (type AB); the resulting T cell reaction of A versus B is inhibited by consecutive treatment. Extent of inhibition can be evaluated after transfer of the pretreated AA material onto AB recipients by calculation of remaining GvH reactivity, if compared to adequate control tranfers. In this model the target animal for T cell reactivity (the AB recipient) remains untouched from immunosuppressive regimen. | [
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|
PMID:11100 | Antibody response to phosphorylcholine in vitro. II. Analysis of T-dependent and T-independent responses. | Spleen cells from BALB/c mice primed with keyhole limpet hemocyanin (KLH), were stimulated with heat-killed vaccine of rough Pneumococcus pneumoniae R36A (Pn) and/or phosphorylcholine (PC)-coupled KLH to induce an anti-PC response in vitro. The response to PC-KLH was found to be T-dependent while it is T-independent to Pn. The antibodies induced with either antigen had similar avidity and expressed the TEPC 15 idiotype exclusively; thus T cell involvement in the response to PC-KLH failed to alter these parameters of the anti-PC response. At the precursor cell level, Pn induced small clones with an average size of 10 plaque-forming cells (PFC), whereas PC-KLH gave rise to larger clones of 40-50 PFC. This difference in the proliferative potential of PC precursor B cells hinted at the possibility that Pn and PC-KLH were stimulating different precursors. This was corroborated by the observation that a) when Pn and PC-KLH were added to the same cultures a synergistic effect was seen, i.e. the number of plaques was greater than the sum of the responses induced by each antigen, and b) in microcultures, under conditions limiting B cells only, Pn plus PC-KLH induced a higher fraction of responding wells than either antigen on its own. We postulate that Pn and PC-KLH stimulate subpopulations of PC precursor cells which are T-independent and T-dependent, respectively. | Antibody response to phosphorylcholine in vitro. II. Analysis of T-dependent and T-independent responses. Spleen cells from BALB/c mice primed with keyhole limpet hemocyanin (KLH), were stimulated with heat-killed vaccine of rough Pneumococcus pneumoniae R36A (Pn) and/or phosphorylcholine (PC)-coupled KLH to induce an anti-PC response in vitro. The response to PC-KLH was found to be T-dependent while it is T-independent to Pn. The antibodies induced with either antigen had similar avidity and expressed the TEPC 15 idiotype exclusively; thus T cell involvement in the response to PC-KLH failed to alter these parameters of the anti-PC response. At the precursor cell level, Pn induced small clones with an average size of 10 plaque-forming cells (PFC), whereas PC-KLH gave rise to larger clones of 40-50 PFC. This difference in the proliferative potential of PC precursor B cells hinted at the possibility that Pn and PC-KLH were stimulating different precursors. This was corroborated by the observation that a) when Pn and PC-KLH were added to the same cultures a synergistic effect was seen, i.e. the number of plaques was greater than the sum of the responses induced by each antigen, and b) in microcultures, under conditions limiting B cells only, Pn plus PC-KLH induced a higher fraction of responding wells than either antigen on its own. We postulate that Pn and PC-KLH stimulate subpopulations of PC precursor cells which are T-independent and T-dependent, respectively. | [
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|
PMID:11101 | Mechanisms by which hapten conjugates of pneumococcal polysaccharide interfere with the challenge of anti-hapten memory cells. | Incubation of trinitrophenylated hemocyanin (TNP-KLH)-primed spleen cells with microgram amounts of 2,4-dinitrophenyl (DNP) or 2,4,6-trinitrophenyl (TNP) conjugates of pneumococcal polysaccharide type 3 (SIII) for as little as 5 min at 4 degrees C results in a specific "block" of the 19 S and 7 S adoptive memory response to TNP-KLH. This hapten-SIII-induced block of anti-hapten memory B cell responsiveness seems to be an example of specific receptor blockade. The block is specific and can be prevented by simultaneous incubation of the primed cells with hapten-protein conjugates which presumably compete with the hapten-polysaccharide for attachment to the B cell surface via anti-hapten Ig receptors. Removal via capping of these Ig receptors by exposure of TNP-KLH-primed memory cells to rabbit anti-mouse Fab serum for 45 min at 37 degrees C renders these cells refractory to the blocking effect of hapten-SIII. Once the hapten-SIII has attached to the memory cells, these blocked cells can be "rescued" (i.e. returned to a state of responsiveness) by incubating these cells with either mouse anti-SIII at 37 degrees C or rabbit anti-DNP serum at 4 degrees C. Since a papain digest of the IgG fraction of rabbit anti-DNP did not rescue the cells while the intact IgG did, a capping off of the TNP-SIII was proposed as the mechanims for this return to responsiveness of the hitherto blocked cells. A rescue was not seen by treatment of recipient mice with such B cell mitogens as dextran sulfate, endotoxin or purified protein derivative of tuberculin. | Mechanisms by which hapten conjugates of pneumococcal polysaccharide interfere with the challenge of anti-hapten memory cells. Incubation of trinitrophenylated hemocyanin (TNP-KLH)-primed spleen cells with microgram amounts of 2,4-dinitrophenyl (DNP) or 2,4,6-trinitrophenyl (TNP) conjugates of pneumococcal polysaccharide type 3 (SIII) for as little as 5 min at 4 degrees C results in a specific "block" of the 19 S and 7 S adoptive memory response to TNP-KLH. This hapten-SIII-induced block of anti-hapten memory B cell responsiveness seems to be an example of specific receptor blockade. The block is specific and can be prevented by simultaneous incubation of the primed cells with hapten-protein conjugates which presumably compete with the hapten-polysaccharide for attachment to the B cell surface via anti-hapten Ig receptors. Removal via capping of these Ig receptors by exposure of TNP-KLH-primed memory cells to rabbit anti-mouse Fab serum for 45 min at 37 degrees C renders these cells refractory to the blocking effect of hapten-SIII. Once the hapten-SIII has attached to the memory cells, these blocked cells can be "rescued" (i.e. returned to a state of responsiveness) by incubating these cells with either mouse anti-SIII at 37 degrees C or rabbit anti-DNP serum at 4 degrees C. Since a papain digest of the IgG fraction of rabbit anti-DNP did not rescue the cells while the intact IgG did, a capping off of the TNP-SIII was proposed as the mechanims for this return to responsiveness of the hitherto blocked cells. A rescue was not seen by treatment of recipient mice with such B cell mitogens as dextran sulfate, endotoxin or purified protein derivative of tuberculin. | [
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|
PMID:11102 | Suprression of local graft-versus-host reactions by mouse fetal and newborn spleen cells. | Fetal splenic but not thymic lymphocytes significantly reduce the ability of parenteral adult spleen cells to elicit local graft-versus-host reactions in F1 recipients. This suppressive activity wanes early after birth. There is no requirement for histo compatibility between reacting and suppressor cells. | Suprression of local graft-versus-host reactions by mouse fetal and newborn spleen cells. Fetal splenic but not thymic lymphocytes significantly reduce the ability of parenteral adult spleen cells to elicit local graft-versus-host reactions in F1 recipients. This suppressive activity wanes early after birth. There is no requirement for histo compatibility between reacting and suppressor cells. | [
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|
PMID:11103 | In vitro studies on smooth muscle of the human renal pelvis. | Isolated segments of human renal pelvis were studied by an isometric technique. Increases in tension following the addition of adrenaline, noradrenaline and phenylephrine were shown to be mediated via alpha-adrenoceptors. Similar responses to acetylcholine were demonstrated to be due to muscarinic receptor stimulation. Specimens responded to transmural electrical stimulation only when the pulse width was greater than 4 msec, and such responses were unaffected by pretreatment with tetrodotoxin, phentolamine and atropine. These experiments suggest that there is no effective innervation of the receptor sites identified, and hence that renal pelvis motility in vivo is not amenable to regulation by the autonomic nervous system. | In vitro studies on smooth muscle of the human renal pelvis. Isolated segments of human renal pelvis were studied by an isometric technique. Increases in tension following the addition of adrenaline, noradrenaline and phenylephrine were shown to be mediated via alpha-adrenoceptors. Similar responses to acetylcholine were demonstrated to be due to muscarinic receptor stimulation. Specimens responded to transmural electrical stimulation only when the pulse width was greater than 4 msec, and such responses were unaffected by pretreatment with tetrodotoxin, phentolamine and atropine. These experiments suggest that there is no effective innervation of the receptor sites identified, and hence that renal pelvis motility in vivo is not amenable to regulation by the autonomic nervous system. | [
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PMID:11104 | Differential response control by isopropamide: a peripherally induced discriminative cue. | The peripherally acting anticholinergic drug isopropamide (0.02 mg/kg s.c.) is shown to produce a discriminative stimulus complex in rats. In rats trained to discriminate isopropamide from saline, dexetimide and methylscopolamine were generalized with isopropamide treatment. The results indicate that drug discrimination learning does not necessarily require a central drug action. | Differential response control by isopropamide: a peripherally induced discriminative cue. The peripherally acting anticholinergic drug isopropamide (0.02 mg/kg s.c.) is shown to produce a discriminative stimulus complex in rats. In rats trained to discriminate isopropamide from saline, dexetimide and methylscopolamine were generalized with isopropamide treatment. The results indicate that drug discrimination learning does not necessarily require a central drug action. | [
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PMID:11105 | Antiarrhythmic action of a new beta-adrenergic blocking agent, 6-(2-hydroxy-3-isopropylaminopropyloxy)-benzothiazole succinate (KF-577), compared with that of propranolol. | The antiarrhythmic activity of a new beta-adrenergic blocking agent, 6-(2-hydroxy-3-isopropylaminopropyloxy)-benzothiazole succinate (KF-577), was compared with that of propranolol. KF-577 antagonized ouabain-induced arrhythmias in normal and bilaterally vagotomized guinea pigs; its antagonistic activity was equal to that of propranolol. Reserpinization greatly reduced ouabain intoxication and neither of the two beta-blockers produced further reduction. Aconitine-induced arrhythmias in rats were not antagonized by the two agents. In intact guinea pigs, the reduction of ouabain intoxication by both beta-blockers could not exceed that produced by simulataneous infusion of KCl, and vice versa. In isolated guinea pig atria, propranolol was about 10 times more effective than KF-577 in reducing the ouabain intoxication. The antiaarhythmic activity of KF-577 paralleled its beta-blocking activity in the isolated preparations but not in the intact animals. | Antiarrhythmic action of a new beta-adrenergic blocking agent, 6-(2-hydroxy-3-isopropylaminopropyloxy)-benzothiazole succinate (KF-577), compared with that of propranolol. The antiarrhythmic activity of a new beta-adrenergic blocking agent, 6-(2-hydroxy-3-isopropylaminopropyloxy)-benzothiazole succinate (KF-577), was compared with that of propranolol. KF-577 antagonized ouabain-induced arrhythmias in normal and bilaterally vagotomized guinea pigs; its antagonistic activity was equal to that of propranolol. Reserpinization greatly reduced ouabain intoxication and neither of the two beta-blockers produced further reduction. Aconitine-induced arrhythmias in rats were not antagonized by the two agents. In intact guinea pigs, the reduction of ouabain intoxication by both beta-blockers could not exceed that produced by simulataneous infusion of KCl, and vice versa. In isolated guinea pig atria, propranolol was about 10 times more effective than KF-577 in reducing the ouabain intoxication. The antiaarhythmic activity of KF-577 paralleled its beta-blocking activity in the isolated preparations but not in the intact animals. | [
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PMID:11106 | Effects of drugs on the formation of homovanillic acid in the rat retina. | Homovanillic acid (HVA) levels were measured in the eye and the corpus striatum of rats under normal conditions and after different drug treatments. Neuroleptic agents such as clozapine, cis-flupenthixol and haloperidol induced comparable increases in HVA levels, whereas the non-neuroleptic trans-isomer of flupenthixol was inactive in both structures. Apomorphine decreased HVA levels in the retina and the corpus striatum, while amphetamine induced a decreased HVA formation in the retina and did not change levels of HVA in the corpus striatum. Probenecid caused a similar percentage rise of HVA in both structures. Morphine and oxotremorine induced a rise in HVA levels in the corpus striatum but not in retinal samples. | Effects of drugs on the formation of homovanillic acid in the rat retina. Homovanillic acid (HVA) levels were measured in the eye and the corpus striatum of rats under normal conditions and after different drug treatments. Neuroleptic agents such as clozapine, cis-flupenthixol and haloperidol induced comparable increases in HVA levels, whereas the non-neuroleptic trans-isomer of flupenthixol was inactive in both structures. Apomorphine decreased HVA levels in the retina and the corpus striatum, while amphetamine induced a decreased HVA formation in the retina and did not change levels of HVA in the corpus striatum. Probenecid caused a similar percentage rise of HVA in both structures. Morphine and oxotremorine induced a rise in HVA levels in the corpus striatum but not in retinal samples. | [
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PMID:11107 | Relationship between the prevention of rat gastric erosions and the inhibition of acid secretion by prostaglandins. | The formation of gastric mucosal erosions induced by indomethacin in the rat was inhibited in a time- and dose-dependent manner by antisecretory prostaglandins, the methyl analogues of PGE2 being 400 times as active as the parent prostaglandin. PGA2, a methyl analogue of PGF2alpha and the H2-receptor antagonist metiamide, also inhibited erosion formation. There was a variable relationship between the doses required to inhibit erosions and to inhibit gastric acid secretion. In the anaesthetised rat, the low incidence of erosions with indomethacin was markedly increased by concurrent gastric perfusion with acid saline and taurocholate. This mucosal damage was inhibited by the methyl analogues of PGE2, suggesting protective actions on the mucosa other than inhibition of acid secretion. | Relationship between the prevention of rat gastric erosions and the inhibition of acid secretion by prostaglandins. The formation of gastric mucosal erosions induced by indomethacin in the rat was inhibited in a time- and dose-dependent manner by antisecretory prostaglandins, the methyl analogues of PGE2 being 400 times as active as the parent prostaglandin. PGA2, a methyl analogue of PGF2alpha and the H2-receptor antagonist metiamide, also inhibited erosion formation. There was a variable relationship between the doses required to inhibit erosions and to inhibit gastric acid secretion. In the anaesthetised rat, the low incidence of erosions with indomethacin was markedly increased by concurrent gastric perfusion with acid saline and taurocholate. This mucosal damage was inhibited by the methyl analogues of PGE2, suggesting protective actions on the mucosa other than inhibition of acid secretion. | [
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|
PMID:11108 | Centrally induced reduction in sympathetic tone-a postsynaptic alpha-adrenoceptor-stimulating action of imidazolines. | Naphazoline or oxymetazoline (both 30 mug/kg) were injected into the cisterna magna of anaesthetized cats and reduced blood pressure, heart rate and the electrical discharge rate of small fibre bundles of the preganglionic sympathetic splanchnic nerve. Cats were depleted of endogenous noradrenaline by pretreatment with reserpine (5 mg/kg, 18 h) and alpha-methyl-p-tyrosine (twice 300 mg/kg, 18 and 2 h). In these animals, intracisternal injection of 30 mug/kg oxymetazoline exerted a decrease of sympathetic discharges similar to that described for non-pretreated animals. In noradrenaline-depleted cats intracisternal injection of 1 mug/kg clonidine also decreased the sympathetic discharges. It is concluded that these imidazolines exert their sympathoinhibitory and cardiovascular effects by stimulation of postsynaptic alpha-adrenoceptors in the CNS. | Centrally induced reduction in sympathetic tone-a postsynaptic alpha-adrenoceptor-stimulating action of imidazolines. Naphazoline or oxymetazoline (both 30 mug/kg) were injected into the cisterna magna of anaesthetized cats and reduced blood pressure, heart rate and the electrical discharge rate of small fibre bundles of the preganglionic sympathetic splanchnic nerve. Cats were depleted of endogenous noradrenaline by pretreatment with reserpine (5 mg/kg, 18 h) and alpha-methyl-p-tyrosine (twice 300 mg/kg, 18 and 2 h). In these animals, intracisternal injection of 30 mug/kg oxymetazoline exerted a decrease of sympathetic discharges similar to that described for non-pretreated animals. In noradrenaline-depleted cats intracisternal injection of 1 mug/kg clonidine also decreased the sympathetic discharges. It is concluded that these imidazolines exert their sympathoinhibitory and cardiovascular effects by stimulation of postsynaptic alpha-adrenoceptors in the CNS. | [
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PMID:11109 | Contribution of granulocytopenia to endotoxin sensitivity of mice irradiated or undergoing graft-versus-host reaction. | Animals compromised by irradiation or graft-versus-host reaction (GVHR) are highly sensitive to endotoxin (ET). In order to determine the causes of the increased sensitivity of compromised mice, we studied alterations of hepatic (central) and bloodborne (peripheral) ET clearance processes. We observed that increased sensitivity to ET, as determined by mortality, occurred shortly after irradiation and correlated with granulocytopenia nd thrombocytopenia rather than with impairment of liver function. The involvement of granulocytes in ET clearance was indicated by injection of 51Cr-ET suspended in whole blood. By comparison with clearance of ET injected with saline or plasma, greater amounts of 51Cr-ET were sequestered in peripheral organs than in the liver. Similar results were obtained when Cr-ET in whole blood was perfused through a rat liver. ET clearance was enhanced 50% over that seen in M-199, plasma, or platelet-rich plasma. It was also found that intestinal ET contributes to mortality of granulocytopenic-thrombocytopenic mice. This was supported by the observation that bacteriologically decontaminated, irradiated animals were eight times more resistant to challenge with ET than were conventional animals. Thus, an important aspect of increased sensitivity to ET, in comprised mice, is defective peripheral clearance. | Contribution of granulocytopenia to endotoxin sensitivity of mice irradiated or undergoing graft-versus-host reaction. Animals compromised by irradiation or graft-versus-host reaction (GVHR) are highly sensitive to endotoxin (ET). In order to determine the causes of the increased sensitivity of compromised mice, we studied alterations of hepatic (central) and bloodborne (peripheral) ET clearance processes. We observed that increased sensitivity to ET, as determined by mortality, occurred shortly after irradiation and correlated with granulocytopenia nd thrombocytopenia rather than with impairment of liver function. The involvement of granulocytes in ET clearance was indicated by injection of 51Cr-ET suspended in whole blood. By comparison with clearance of ET injected with saline or plasma, greater amounts of 51Cr-ET were sequestered in peripheral organs than in the liver. Similar results were obtained when Cr-ET in whole blood was perfused through a rat liver. ET clearance was enhanced 50% over that seen in M-199, plasma, or platelet-rich plasma. It was also found that intestinal ET contributes to mortality of granulocytopenic-thrombocytopenic mice. This was supported by the observation that bacteriologically decontaminated, irradiated animals were eight times more resistant to challenge with ET than were conventional animals. Thus, an important aspect of increased sensitivity to ET, in comprised mice, is defective peripheral clearance. | [
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PMID:11110 | Mitigation of Graft-versus-host disease in mice with xenogeneic antithymocyte serum and complement. | In vitro treatment of parental C57BL/6 lymphohematopoietic cell grafts with unabsorbed guinea pig anti-mouse thymocyte serum (ATS) and guinea pig complement (GPC), prior to inoculation into lethally irradiated B6D2F hybrid hosts, has proven to be of value in terms of mitigating graft-versus-host disease (GvHD). However, the beneficial effect of such a pregrafting procedure is limited to the prevention of acute GvHD. The late GvHD remains a continuing problem, and is probably due to the graft-versus-host activity (GvHA) of newly produced nontolerant lymphocytes from lymphoid precursors resistant to ATS. Possible ways to render these precursors sensitive to ATS and complement are discussed. The potential significance of thymic hormones and cyclic AMP in achieving this is emphasized. | Mitigation of Graft-versus-host disease in mice with xenogeneic antithymocyte serum and complement. In vitro treatment of parental C57BL/6 lymphohematopoietic cell grafts with unabsorbed guinea pig anti-mouse thymocyte serum (ATS) and guinea pig complement (GPC), prior to inoculation into lethally irradiated B6D2F hybrid hosts, has proven to be of value in terms of mitigating graft-versus-host disease (GvHD). However, the beneficial effect of such a pregrafting procedure is limited to the prevention of acute GvHD. The late GvHD remains a continuing problem, and is probably due to the graft-versus-host activity (GvHA) of newly produced nontolerant lymphocytes from lymphoid precursors resistant to ATS. Possible ways to render these precursors sensitive to ATS and complement are discussed. The potential significance of thymic hormones and cyclic AMP in achieving this is emphasized. | [
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|
PMID:11111 | Acid mucopolysaccharides in fibroblast cultures. 1. Influence of cell density, pH-value and lactate concentration on the MPS distribution pattern. | From cells and culture media of embryonic rat fibroblasts (1st subculture) the acid mucopolysaccharides were isolated and fractionated. The per cent calculation of the 6 fractions was based on the content of glucuronic acid. The cultures were maintained as follows: 0.5 X 10(6) to 3 X 10(6) cells were examined in Demeter flasks at pH 7.4 or 6.6; lactate concentration was enhanced to 100 mg%. The amount of each fraction was correlated with the cell density (linear regression). The pH-value and lactate concentration in connection with cell density proved to be important factors in the modification of the MPS distribution pattern. | Acid mucopolysaccharides in fibroblast cultures. 1. Influence of cell density, pH-value and lactate concentration on the MPS distribution pattern. From cells and culture media of embryonic rat fibroblasts (1st subculture) the acid mucopolysaccharides were isolated and fractionated. The per cent calculation of the 6 fractions was based on the content of glucuronic acid. The cultures were maintained as follows: 0.5 X 10(6) to 3 X 10(6) cells were examined in Demeter flasks at pH 7.4 or 6.6; lactate concentration was enhanced to 100 mg%. The amount of each fraction was correlated with the cell density (linear regression). The pH-value and lactate concentration in connection with cell density proved to be important factors in the modification of the MPS distribution pattern. | [
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|
PMID:11112 | Acid mucopolysaccharides in fibroblast cultures. 2. 35S-sulfate incorporation kinetics in dependence on pH-value and cell density. | Cultures of embryonic rat fibroblasts were incubated with 35S-sulfate at pH 6.6 and 7.4 (Eagle basal medium plus HEPES buffer) for 12 to 48 hours. The acid mucopolysaccharides were isolated and fractionated after the method of SVEJCAR and ROBERTSON. Sulfate incorporation was determined by liquid scintillation counting. | Acid mucopolysaccharides in fibroblast cultures. 2. 35S-sulfate incorporation kinetics in dependence on pH-value and cell density. Cultures of embryonic rat fibroblasts were incubated with 35S-sulfate at pH 6.6 and 7.4 (Eagle basal medium plus HEPES buffer) for 12 to 48 hours. The acid mucopolysaccharides were isolated and fractionated after the method of SVEJCAR and ROBERTSON. Sulfate incorporation was determined by liquid scintillation counting. | [
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|
PMID:11114 | Allantoinase in the marine polychaete Eudistylia vancouveri. | Allantoinase, an enzyme in the purine-urea cycle, was found in Eudistylia vancouveri (Polychaeta). The enzyme had a pH optimum at 7.6. The Km was 0.012 M allantoin, and the Arrhenius energy of activation was 12.6 to 14.6 kcal/mol. | Allantoinase in the marine polychaete Eudistylia vancouveri. Allantoinase, an enzyme in the purine-urea cycle, was found in Eudistylia vancouveri (Polychaeta). The enzyme had a pH optimum at 7.6. The Km was 0.012 M allantoin, and the Arrhenius energy of activation was 12.6 to 14.6 kcal/mol. | [
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|
PMID:11116 | Influence of the trypsin activity by the side chain of arginine homologues. | The N-alpha-tosyl-p-nitroanilides of homoarginine and of the two shorter arginine homologues were synthesized. These compounds behave as specific, chromogenic substrates for trypsin. | Influence of the trypsin activity by the side chain of arginine homologues. The N-alpha-tosyl-p-nitroanilides of homoarginine and of the two shorter arginine homologues were synthesized. These compounds behave as specific, chromogenic substrates for trypsin. | [
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|
PMID:11117 | Beta-adrenergic receptors in rat myocardium: direct detection by a new fluorescent beta-blocker. | A new fluorescent beta-blocker, 9-amino-acridin propranolol (9-AAP), was administered i.v. to rats. Multiple fluorescent 9-AAP binding sites were observed on cardiac muscle cells in frozen sections. Intensity and density of cardiac 9-AAP fluorescence were markedly reduced following pretreatment with (+/-)- and (-)-propranolol but not with (+)-propranolol. Our findings suggest that 9-AAP may label beta-adrenergic receptor sites in rat myocardium. | Beta-adrenergic receptors in rat myocardium: direct detection by a new fluorescent beta-blocker. A new fluorescent beta-blocker, 9-amino-acridin propranolol (9-AAP), was administered i.v. to rats. Multiple fluorescent 9-AAP binding sites were observed on cardiac muscle cells in frozen sections. Intensity and density of cardiac 9-AAP fluorescence were markedly reduced following pretreatment with (+/-)- and (-)-propranolol but not with (+)-propranolol. Our findings suggest that 9-AAP may label beta-adrenergic receptor sites in rat myocardium. | [
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|
PMID:11118 | Histamine-induced hypotension modified by H1 and H2 antagonists in the monkey (Macaca mulatta). | Blocking H2 receptors with burimamide in the dose used (20 mg/kg) approximately doubles the amount of histamine needed to produce the same effect as seen when H1 antagonists (chlorpheniramine or mepyramine) are used alone. The Kz ratios for chlorpheniramine-chlorpheniramine plus burimamide are 117-204 and for mepyramine-mepyramine phus burimamide are 200-478. H1 and H2 receptors, in the monkey, when stimulated, both cause cardiovascular responses in the same direction. | Histamine-induced hypotension modified by H1 and H2 antagonists in the monkey (Macaca mulatta). Blocking H2 receptors with burimamide in the dose used (20 mg/kg) approximately doubles the amount of histamine needed to produce the same effect as seen when H1 antagonists (chlorpheniramine or mepyramine) are used alone. The Kz ratios for chlorpheniramine-chlorpheniramine plus burimamide are 117-204 and for mepyramine-mepyramine phus burimamide are 200-478. H1 and H2 receptors, in the monkey, when stimulated, both cause cardiovascular responses in the same direction. | [
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|
PMID:11119 | The oxygen-linked hydrogen ion binding (the Haldane coefficient) of bovine hemoglobin. | The Haldane coefficient (the amount of the oxygen-linked hydrogen ion binding of hemoglobin) was determined in bovine erythrolysate (Hb concentration equals 13.5 mM) by means of the differential titration method with varying PCO2 from 0 to 74 mm Hg and pH from 6.0 to 8.5 at 37 degrees C. The maximum value of the coefficient was found to be 0.49 mM per mM Hb at PCO2 equals 0 and pH 7.20. With increasing of PCO2, the coefficient became smaller in all ranges of pH studied. The coefficient under the conditions of pH 7.20 and PCO2 equals 45 mm Hg that are normally prevailing in the interior of bovine erythrocytes was 0.31. | The oxygen-linked hydrogen ion binding (the Haldane coefficient) of bovine hemoglobin. The Haldane coefficient (the amount of the oxygen-linked hydrogen ion binding of hemoglobin) was determined in bovine erythrolysate (Hb concentration equals 13.5 mM) by means of the differential titration method with varying PCO2 from 0 to 74 mm Hg and pH from 6.0 to 8.5 at 37 degrees C. The maximum value of the coefficient was found to be 0.49 mM per mM Hb at PCO2 equals 0 and pH 7.20. With increasing of PCO2, the coefficient became smaller in all ranges of pH studied. The coefficient under the conditions of pH 7.20 and PCO2 equals 45 mm Hg that are normally prevailing in the interior of bovine erythrocytes was 0.31. | [
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|
PMID:11120 | Activation of sustained sympathetic vasodilatation in dog by spinal cord stimulation. | Electrical stimulation in lateral sites of the upper cervical spinal cord evoked vasodilatation after adrenergic blockade. Sympathetic fibres mediating sustained vasodilatation were shown to be separate from adrenergic sympathetic fibres since the adrenergic vasoconstrictor response in the paw evoked by vasomotor stimulation in the medulla was not reversed to vasodilatation after bretylium. | Activation of sustained sympathetic vasodilatation in dog by spinal cord stimulation. Electrical stimulation in lateral sites of the upper cervical spinal cord evoked vasodilatation after adrenergic blockade. Sympathetic fibres mediating sustained vasodilatation were shown to be separate from adrenergic sympathetic fibres since the adrenergic vasoconstrictor response in the paw evoked by vasomotor stimulation in the medulla was not reversed to vasodilatation after bretylium. | [
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|
PMID:11121 | Effects of morphine administration on cerebellar guanosine 3',5'-monophosphate. | An increase in mouse cerebellar C-GMP levels during acute morphine treatment was observed, which was possibly related to the decrease in C-GMP phosphodiesterase levels also observed in acute treatment. Chronic treatment lowered C-GMP levels as did abrupt withdrawal without naloxone. | Effects of morphine administration on cerebellar guanosine 3',5'-monophosphate. An increase in mouse cerebellar C-GMP levels during acute morphine treatment was observed, which was possibly related to the decrease in C-GMP phosphodiesterase levels also observed in acute treatment. Chronic treatment lowered C-GMP levels as did abrupt withdrawal without naloxone. | [
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|
PMID:11122 | [Effect of liver damage by thioacetamide on microsomal aromatization of testosterone in rats (author's transl)]. | Rat liver microsomes, NADPH-regenerating system, and 1beta, 2beta-3H-testosterone have been incubated in vitro. The loss of tritium from the steroid, associated with aromatization of testosterone, was linear with time for 20 min and required NADPH. Pre-treatment of the rats with thioacetamide raised the liberation of tritium from 1beta, 2beta-3H-testosterone. The results suggest that liver damage by thioacetamide in rats may give rise to increased aromatization of testosterone. | [Effect of liver damage by thioacetamide on microsomal aromatization of testosterone in rats (author's transl)]. Rat liver microsomes, NADPH-regenerating system, and 1beta, 2beta-3H-testosterone have been incubated in vitro. The loss of tritium from the steroid, associated with aromatization of testosterone, was linear with time for 20 min and required NADPH. Pre-treatment of the rats with thioacetamide raised the liberation of tritium from 1beta, 2beta-3H-testosterone. The results suggest that liver damage by thioacetamide in rats may give rise to increased aromatization of testosterone. | [
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|
PMID:11136 | Reduction of the graft-versus-host reactivity of mouse and rat spleen cells by 5alpha-androstane-3,17-dione. | The local graft-versus-host reaction as evaluated by the popliteal lymph node enlargement was used for studying the immunosuppressive potency of placental steroid 5alpha-androstane-3,17-dione. Spleen cells from virgin female mice and rats injected subcutaneously with this steroid compound produced in appropriate F1 recipients a significantly weaker reaction (P less than 0.001) than spleen cells from untreated or medium-treated control animals. On the other hand, the pretreatment of cell donors either with 5beta-androstane-3,17-dione or testosterone, the compounds which are not present in the mouse and rat placenta, did not influence the normal graft-versus-host reactivity. | Reduction of the graft-versus-host reactivity of mouse and rat spleen cells by 5alpha-androstane-3,17-dione. The local graft-versus-host reaction as evaluated by the popliteal lymph node enlargement was used for studying the immunosuppressive potency of placental steroid 5alpha-androstane-3,17-dione. Spleen cells from virgin female mice and rats injected subcutaneously with this steroid compound produced in appropriate F1 recipients a significantly weaker reaction (P less than 0.001) than spleen cells from untreated or medium-treated control animals. On the other hand, the pretreatment of cell donors either with 5beta-androstane-3,17-dione or testosterone, the compounds which are not present in the mouse and rat placenta, did not influence the normal graft-versus-host reactivity. | [
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|
PMID:11137 | The sensitivity of chromatin from thymuses and spleens of irradiated mice to alkaline solutions. | Increasing amounts of DNA and proteins are released from the suspensions of chromatin from thymuses and spleens of irradiated mice (6 hours after 600R whole-body) by the action of alkaline solutions (pH 8-10) at physiological ionic strengths. The suspension of chromatin from normal tissues releases in this pH range only a small amount of proteins and negligible amount of DNA. The behaviour of liver and kidney chromatin to alkaline solutions shows no difference between normal and irradiated tissues. The time of onset and dose relation of the increased sensitivity of thymus and spleen chromatin from irradiated mice to alkaline solutions show a similar course as earlier described signs of postirradiation damage to chromatin of these tissues. | The sensitivity of chromatin from thymuses and spleens of irradiated mice to alkaline solutions. Increasing amounts of DNA and proteins are released from the suspensions of chromatin from thymuses and spleens of irradiated mice (6 hours after 600R whole-body) by the action of alkaline solutions (pH 8-10) at physiological ionic strengths. The suspension of chromatin from normal tissues releases in this pH range only a small amount of proteins and negligible amount of DNA. The behaviour of liver and kidney chromatin to alkaline solutions shows no difference between normal and irradiated tissues. The time of onset and dose relation of the increased sensitivity of thymus and spleen chromatin from irradiated mice to alkaline solutions show a similar course as earlier described signs of postirradiation damage to chromatin of these tissues. | [
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|
PMID:11138 | Some biological properties of mouse spleen cells fractionated by the adherence of Sephadex G 25 and glass bead columns. | The possibilities of separation of haemopoietic cells from lymphocytes capable of eliciting the graft-versus-host reaction through column chromatography were investigated. Strain-A mouse spleen cells were fractionated into the adherent and non-adherent fraction on Sephadex G-25, glass bead columns and glass beads coated with antibody against mouse globulin. Increased numbers of cells forming haemopoietic colonies were found in the cell fraction which did not adhere to the antibody-coated glass beads and in cells reversibly adhering to glass beads. No significant decrease in local graft-versus-host reaction was found in any fraction obtained, and the prolonged survival of irradiated semiallogeneic recipients was observed in both fractions obtained on Sephadex G-25 columns. | Some biological properties of mouse spleen cells fractionated by the adherence of Sephadex G 25 and glass bead columns. The possibilities of separation of haemopoietic cells from lymphocytes capable of eliciting the graft-versus-host reaction through column chromatography were investigated. Strain-A mouse spleen cells were fractionated into the adherent and non-adherent fraction on Sephadex G-25, glass bead columns and glass beads coated with antibody against mouse globulin. Increased numbers of cells forming haemopoietic colonies were found in the cell fraction which did not adhere to the antibody-coated glass beads and in cells reversibly adhering to glass beads. No significant decrease in local graft-versus-host reaction was found in any fraction obtained, and the prolonged survival of irradiated semiallogeneic recipients was observed in both fractions obtained on Sephadex G-25 columns. | [
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PMID:11140 | Dopamine as a possible neurotransmitter in gastric relaxation. | In dogs with gastric fistulas, intragastric pressure was measured with a flaccid ballon containing 500 ml of water. Graded doses of dopamine caused graded decreases in intragastric pressure. The effect was blocked by pimozide or by metoclopramide but was not significantly affected by phenoxybenzamine, propranolol, guanethidine, or FLA-63 (a beta-hydroxylase inhibitor). Pretreatment with metoclopramide or with pimozide shifted the volume-pressure diagram of the stomach to the left; that is, at any given volume the pressure was greater after than before these drugs. In dogs with vagally innervated gastric pouches and gastric fistulas, feeding for 1 min (while allowing the food to leave the stomach through the gastric fistula) caused a prompt decrease in pressure in the pouch that lasted for about 5 min. Pretreatment with metoclopramide decreased the magnitude and duration of this receptive relaxation. It is concluded that these findings are consistent with (but do not establish) the hypothesis that dopamine is the neurotransmitter for receptive relaxation of the stomach, because dopamine mimics receptive relaxation, and dopamine antagonists partially block reflexly induced receptive relaxation. | Dopamine as a possible neurotransmitter in gastric relaxation. In dogs with gastric fistulas, intragastric pressure was measured with a flaccid ballon containing 500 ml of water. Graded doses of dopamine caused graded decreases in intragastric pressure. The effect was blocked by pimozide or by metoclopramide but was not significantly affected by phenoxybenzamine, propranolol, guanethidine, or FLA-63 (a beta-hydroxylase inhibitor). Pretreatment with metoclopramide or with pimozide shifted the volume-pressure diagram of the stomach to the left; that is, at any given volume the pressure was greater after than before these drugs. In dogs with vagally innervated gastric pouches and gastric fistulas, feeding for 1 min (while allowing the food to leave the stomach through the gastric fistula) caused a prompt decrease in pressure in the pouch that lasted for about 5 min. Pretreatment with metoclopramide decreased the magnitude and duration of this receptive relaxation. It is concluded that these findings are consistent with (but do not establish) the hypothesis that dopamine is the neurotransmitter for receptive relaxation of the stomach, because dopamine mimics receptive relaxation, and dopamine antagonists partially block reflexly induced receptive relaxation. | [
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PMID:11141 | Inhibition of intestinal iron absorption by laundry starch. | The pathogenesis of iron deficiency anemia associated with amylophagia is usually attributed to dietary iron lack. However, large quantities of starch may inhibit intestinal iron absorption. Accordingly, studies were carried out to determine the effect of laundry starch on the intestinal absorption of inorganic and hemoglobin iron. In vitro, laundry starch bound 19 to 80% of the available 59FeSO4 and 34 to 68% of the available 59Fe-hemoglobin. Binding of both forms of iron was pH-dependent, with maximal binding at pH 7.0. In vivo, laundry starch significantly inhibited mucosal uptake of 59FeSO4 from isolated duodenal loops. In nonanemic rats, administration of laundry starch (100 mg) 1 hr before a 100-mug dose of 59FeSO4 significantly decreased the absorption of 59FeSO4, as compared to saline or low iron chow controls (6.2 +/- 2.0 versus 14.9 +/- 2.1 and -1.8 +/- 1.7, respectively, P less than 0.001). In anemic rats the absorption of either a 100-mug dose of 59FeSO4 or a 500-mug dose of 59Fe-hemoglobin was also significantly decreased by prior administration of laundry starch. The data obtained indicated that laundry starch (1) binds appreciable quantities of inorganic and hemoglobin iron in vitro; (2) inhibits the mucosal uptake or inorganic iron by isolated intestinal loops; (3) inhibits the intestinal absorption of inorganic iron in normal nonanemic rats, and (4) blunts the compensatory increase in inorganic and organic iron absorption in anemic rats. | Inhibition of intestinal iron absorption by laundry starch. The pathogenesis of iron deficiency anemia associated with amylophagia is usually attributed to dietary iron lack. However, large quantities of starch may inhibit intestinal iron absorption. Accordingly, studies were carried out to determine the effect of laundry starch on the intestinal absorption of inorganic and hemoglobin iron. In vitro, laundry starch bound 19 to 80% of the available 59FeSO4 and 34 to 68% of the available 59Fe-hemoglobin. Binding of both forms of iron was pH-dependent, with maximal binding at pH 7.0. In vivo, laundry starch significantly inhibited mucosal uptake of 59FeSO4 from isolated duodenal loops. In nonanemic rats, administration of laundry starch (100 mg) 1 hr before a 100-mug dose of 59FeSO4 significantly decreased the absorption of 59FeSO4, as compared to saline or low iron chow controls (6.2 +/- 2.0 versus 14.9 +/- 2.1 and -1.8 +/- 1.7, respectively, P less than 0.001). In anemic rats the absorption of either a 100-mug dose of 59FeSO4 or a 500-mug dose of 59Fe-hemoglobin was also significantly decreased by prior administration of laundry starch. The data obtained indicated that laundry starch (1) binds appreciable quantities of inorganic and hemoglobin iron in vitro; (2) inhibits the mucosal uptake or inorganic iron by isolated intestinal loops; (3) inhibits the intestinal absorption of inorganic iron in normal nonanemic rats, and (4) blunts the compensatory increase in inorganic and organic iron absorption in anemic rats. | [
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PMID:11142 | Effect of serotonin on water and electrolyte transport in the in vivo rabbit small intestine. | The influence of intravenously administered serotonin on water and electrolyte fluxes in the in vivo rabbit jejunum and ileum was examined. Animals were divided into four groups: (1) those receiving saline intravenously while a glucose-free isotonic saline solution perfused the jejunum and ileum; (2) serotonin given intravenously while glucose-free intestinal perfusate was used as in group 1; (3) intravenous saline given while a 10 mM glucose-isotonic saline solution perfused the jejunum and ileum; and (4) intravenous serotonin given while the intestinal perfusate was as in group 3. Serotonin administration resulted in highly significant net secretion of H2O and sodium in both jejunum and ileum in the groups with a glucose-free perfusate. In jejunum, serotonin evoked net water and sodium secretion, whereas controls absorbed water and sodium. In ileum, serotonin significantly enhanced secretion. The addition of glucose to the perfusate completely abolished the serotonin effect. Unidirectional 22Na flux analysis revealed a marked diminution in both mucosal to serosal and serosal to mucosal fluxes in serotonin-treated animals. The decrease in mucosal to serosal flux was greater than the decrease in serosal to mucosal flux, thus explaining the enhanced net secretion observed with serotonin in the groups receiving glucose-free perfusate. In spite of its pronounced effect on water and electrolyte transport, serotonin failed to produce any detectable histological alterations in small bowel mucosa, either by light or electron microscopy. We postulate that serotonin may be an important mediator of the diarrhea so frequently noted in the carcinoid syndrome by virtue of its effects on small intestinal H2O and electrolyte transport. | Effect of serotonin on water and electrolyte transport in the in vivo rabbit small intestine. The influence of intravenously administered serotonin on water and electrolyte fluxes in the in vivo rabbit jejunum and ileum was examined. Animals were divided into four groups: (1) those receiving saline intravenously while a glucose-free isotonic saline solution perfused the jejunum and ileum; (2) serotonin given intravenously while glucose-free intestinal perfusate was used as in group 1; (3) intravenous saline given while a 10 mM glucose-isotonic saline solution perfused the jejunum and ileum; and (4) intravenous serotonin given while the intestinal perfusate was as in group 3. Serotonin administration resulted in highly significant net secretion of H2O and sodium in both jejunum and ileum in the groups with a glucose-free perfusate. In jejunum, serotonin evoked net water and sodium secretion, whereas controls absorbed water and sodium. In ileum, serotonin significantly enhanced secretion. The addition of glucose to the perfusate completely abolished the serotonin effect. Unidirectional 22Na flux analysis revealed a marked diminution in both mucosal to serosal and serosal to mucosal fluxes in serotonin-treated animals. The decrease in mucosal to serosal flux was greater than the decrease in serosal to mucosal flux, thus explaining the enhanced net secretion observed with serotonin in the groups receiving glucose-free perfusate. In spite of its pronounced effect on water and electrolyte transport, serotonin failed to produce any detectable histological alterations in small bowel mucosa, either by light or electron microscopy. We postulate that serotonin may be an important mediator of the diarrhea so frequently noted in the carcinoid syndrome by virtue of its effects on small intestinal H2O and electrolyte transport. | [
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|
PMID:11144 | Lower esophageal sphincter response to oral administration of cimetidine in normal subjects. | Anithistamines that specifically block the gastric and secretory action of histamine have recently been developed. One of these H2-receptor blockers, metiamide, has been found to increase lower esophageal sphincter (LES) pressure in the opossum. Because of reported agranulocytosis with metiamide, another H2-receptor blocking agent, cimetidine, was developed. To determine its effect on LES pressure, 8 normal volunteers received placebo or oral doses of cimetidine (50, 100, 200, and 400 mg) in a random, blinded manner. Indicative of adequate absorption, significant serum levels were achieved with all doses of cimetidine (50 mg = 0.17 mug per ml; 100 mg = 0.33 mug per ml; 200 mg = 0.76 mug per ml; and 400 mg = 1.61 mug per ml). Although these serum levels have been found to produce marked inhibition of gastric acid secretion, no discernible effect was found on LES pressure when compared to placebo. Thus cimetidine does not increase LES pressure. It does not decrease sphincter pressure either and is therefore not contraindicated in patients with reflux esophagitis. | Lower esophageal sphincter response to oral administration of cimetidine in normal subjects. Anithistamines that specifically block the gastric and secretory action of histamine have recently been developed. One of these H2-receptor blockers, metiamide, has been found to increase lower esophageal sphincter (LES) pressure in the opossum. Because of reported agranulocytosis with metiamide, another H2-receptor blocking agent, cimetidine, was developed. To determine its effect on LES pressure, 8 normal volunteers received placebo or oral doses of cimetidine (50, 100, 200, and 400 mg) in a random, blinded manner. Indicative of adequate absorption, significant serum levels were achieved with all doses of cimetidine (50 mg = 0.17 mug per ml; 100 mg = 0.33 mug per ml; 200 mg = 0.76 mug per ml; and 400 mg = 1.61 mug per ml). Although these serum levels have been found to produce marked inhibition of gastric acid secretion, no discernible effect was found on LES pressure when compared to placebo. Thus cimetidine does not increase LES pressure. It does not decrease sphincter pressure either and is therefore not contraindicated in patients with reflux esophagitis. | [
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|
PMID:11145 | The role of histamine receptors in the pathophysiology of gastric mucosal damage. | In four canine Heidenhain pouches the net fluxes of H+ and Na+ have been examined before, during, and after instillation of sodium taurocholate into the pouch. These experiments were conducted in animals given H1 (mepyramine maleate) and H2 (metiamide) histamine antagonists, alone and in combination. Control experiments without antagonists were also conducted. In control experiments, as well as in those using the histamine antagonists separately, the usual sequence of events followed exposure to taurocholate-that is, a gain in the volume of the solution in the pouch and an increase in the fluxes of Na+ and H+ across the mucosa. In experiments in which H1 and H2 histamine antagonists were used in combination, taurocholate had very little effect on the ionic fluxes of H+ and Na+, suggesting that changes in the ionic permeability of the gastric mucosal barrier are mediated by histamine through both H1 and H2 receptor sites. | The role of histamine receptors in the pathophysiology of gastric mucosal damage. In four canine Heidenhain pouches the net fluxes of H+ and Na+ have been examined before, during, and after instillation of sodium taurocholate into the pouch. These experiments were conducted in animals given H1 (mepyramine maleate) and H2 (metiamide) histamine antagonists, alone and in combination. Control experiments without antagonists were also conducted. In control experiments, as well as in those using the histamine antagonists separately, the usual sequence of events followed exposure to taurocholate-that is, a gain in the volume of the solution in the pouch and an increase in the fluxes of Na+ and H+ across the mucosa. In experiments in which H1 and H2 histamine antagonists were used in combination, taurocholate had very little effect on the ionic fluxes of H+ and Na+, suggesting that changes in the ionic permeability of the gastric mucosal barrier are mediated by histamine through both H1 and H2 receptor sites. | [
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|
PMID:11146 | Properties of gastric antrum. III. Selectivity and modification of shunt conductance. | The permselectivity of the paracellular pathway of amphibian (Necturus and bullfrog) antrum was investigated with respect to intracationic selectivity and the K+ and Cl- permeability ratio as a function of mucosal pH. The intracationic selectivity sequence was Rb+ greater than K+ greater than Cs+ greater than Na+ greater than Li+. Both antra showed weak cationic selectivity at pH 7.4, and at pH 4.4 for bullfrog and pH 3.0 for Necturus, the ratio Pk+/P Cl- was unity. At lower mucosal pH the tissues were anion selective. Treatment of the tissue with a water-soluble carbodiimide enhanced anion selectivity at higher pH; carbenoxolone, a weak acid, resulted in maintained cation selectivity at lower pH. These data suggest that carboxyl groups play a role in determining shunt selectivity; the increase in anion selectivity below pH 2.0 suggests that phosphate or sulfate groups could also be involved. | Properties of gastric antrum. III. Selectivity and modification of shunt conductance. The permselectivity of the paracellular pathway of amphibian (Necturus and bullfrog) antrum was investigated with respect to intracationic selectivity and the K+ and Cl- permeability ratio as a function of mucosal pH. The intracationic selectivity sequence was Rb+ greater than K+ greater than Cs+ greater than Na+ greater than Li+. Both antra showed weak cationic selectivity at pH 7.4, and at pH 4.4 for bullfrog and pH 3.0 for Necturus, the ratio Pk+/P Cl- was unity. At lower mucosal pH the tissues were anion selective. Treatment of the tissue with a water-soluble carbodiimide enhanced anion selectivity at higher pH; carbenoxolone, a weak acid, resulted in maintained cation selectivity at lower pH. These data suggest that carboxyl groups play a role in determining shunt selectivity; the increase in anion selectivity below pH 2.0 suggests that phosphate or sulfate groups could also be involved. | [
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|
PMID:11147 | Effect of fundusectomy on serum and antral gastrin levels in rats. | In adult male rats, fundusectomy decreased acid secretion but significantly increased total antral gastrin and both fasting and food-stimulated serum gastrin levels. The rise in fasting serum gastrin could be inhibited by antral acidification, suggesting that decreased acidity caused postfundusectomy hypergastrinemia. The mechanism for the increase in total gastrin in antral tissue is probably the same. These studies provide a useful experimental model for the increasing of antral gastrin and for the production of hypergastrinemia. | Effect of fundusectomy on serum and antral gastrin levels in rats. In adult male rats, fundusectomy decreased acid secretion but significantly increased total antral gastrin and both fasting and food-stimulated serum gastrin levels. The rise in fasting serum gastrin could be inhibited by antral acidification, suggesting that decreased acidity caused postfundusectomy hypergastrinemia. The mechanism for the increase in total gastrin in antral tissue is probably the same. These studies provide a useful experimental model for the increasing of antral gastrin and for the production of hypergastrinemia. | [
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|
PMID:11148 | Postprandial gastric, pancreatic, and biliary response to histamine H2-receptor antagonists active duodenal ulcer. | Histamine H2-receptor antagonists are potentially useful agents in duodenal ulcer and knowledge of their effect on postprandial digestive events will contribute to their clinical application. We studied the effect of 200- and 300-mg doses of cimetidine, an H2-receptor antagonist, taken with an ordinary meal, on gastric, pancreatic, and biliary function. Both doses significantly reduced acid output and its delivery into the duodenum. Gastric secretory volume and pepsin output were less affected. Acid inhibition was related to blood drug levels and was less than that previously found at night in nocturnal fasting studies. As the stomach emptied the food, the gastric pH rose. The fractional gastric emptying rate, pancreatic enzyme, and bile acid outputs were unaltered. Cimetidine taken orally with meals at these doses is a potent gastric antisecretory agent without affecting other postprandial gastric, pancreatic, or biliary functions. | Postprandial gastric, pancreatic, and biliary response to histamine H2-receptor antagonists active duodenal ulcer. Histamine H2-receptor antagonists are potentially useful agents in duodenal ulcer and knowledge of their effect on postprandial digestive events will contribute to their clinical application. We studied the effect of 200- and 300-mg doses of cimetidine, an H2-receptor antagonist, taken with an ordinary meal, on gastric, pancreatic, and biliary function. Both doses significantly reduced acid output and its delivery into the duodenum. Gastric secretory volume and pepsin output were less affected. Acid inhibition was related to blood drug levels and was less than that previously found at night in nocturnal fasting studies. As the stomach emptied the food, the gastric pH rose. The fractional gastric emptying rate, pancreatic enzyme, and bile acid outputs were unaltered. Cimetidine taken orally with meals at these doses is a potent gastric antisecretory agent without affecting other postprandial gastric, pancreatic, or biliary functions. | [
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|
PMID:11149 | Isolation and characterization of four peptide hydrolases from the cytosol of rat intestinal mucosa. | The high speed supernatant fluid prepared from rat intestinal mucosa was subjected to ion-exchange chromatography on diethlaminoethyl-cellulose eluted with a linear gradient of sodium chloride (0 to 0.27 M). Assay of eluted fractions for Phe-Gly hydrolase activity revealed four distinct peaks of enzyme activity. These cytosol enzymes have been designated I, II, III, and IV in order of their elution from the column. Examination of the substrate specificity of the four enzymes by use of 20 mM peptide concentrations indicated the most discriminating substrates for the four enzymes were Leu-Gly-Gly, His-Met, Ser-Phe, and leucine amide, respectively. The mean distribution of the recovered peptide hydrolase activities against these substrates among the four enzymes I, II, III, and IV was 96.1, 1.4, 1.7, and 0.8%, respectively, for Leu-Gly-Gly; 0.6, 96.4, 2.4, and 0.6% for His-Met; 0, 0, 95.8, and 4.2% for Ser-Phe; and 20.8, 19.8, 5.6, and 53.8% for leucine amide. Ion-exchange chromatography resulted in increases in specific activity of 19-, 19-, 46-, and 3.5-fold for enzymes I, II, III, and IV, respectively. The activity of all four enzymes, but especially III and IV, were stabilized by the presence of 150 muM dithioerythritol. Activity of each of the four enzymes was decreased 79 to 100% by 1mM ethylenediaminetetraacetate, HgCl2, 1, 10-phenanthroline, or 0.5 mM p-hydroxymercuribenzoate, except that the activity of enzyme I was decreased only 15% by ethylenediaminetetraacetate. No significant activation of the partially purified enzymes occurred in the presence of 500 muM Zn++, Co++, or Mg++. The four enzymes exhibited distinct pH profiles with optima at 7.5, 7.5, 8.5, and 8.0 for enzymes I, II, III, and IV, respectively. Molecular weights of the four enzymes determined by gel filtration on Sephadex G-200 were 58,500, 74,000, 97,500, and 113,000, respectively. All four enzymes lost more than 85% of their activity after 1 hr at temperatures of 50 degrees C or higher in sodium phosphate buffer, pH 7.0. The Km values determined with the most specific substrates for each enzyme were 0.76, 0.44, 3.82, and 8.3 mM for enzymes I, II, III, and IV, respectively. Recent evidence suggests that a significant amount of some small peptides are absorbed intact and hydrolyzed by cytosol peptide hydrolases. Adequate understanding of the function and control of these intracellular enzymes requires knowledge of the characteristics and substrates specificity of individual enzymes. The study described here demonstrates the presence of at least four cytosol peptide hydrolases with distinct substrate specificities. Substrates almost exclusively hydrolyzed by each of three of the enzymes, and therefore suitable for assay of each of these enzymes in the presence of the others, have been identified. | Isolation and characterization of four peptide hydrolases from the cytosol of rat intestinal mucosa. The high speed supernatant fluid prepared from rat intestinal mucosa was subjected to ion-exchange chromatography on diethlaminoethyl-cellulose eluted with a linear gradient of sodium chloride (0 to 0.27 M). Assay of eluted fractions for Phe-Gly hydrolase activity revealed four distinct peaks of enzyme activity. These cytosol enzymes have been designated I, II, III, and IV in order of their elution from the column. Examination of the substrate specificity of the four enzymes by use of 20 mM peptide concentrations indicated the most discriminating substrates for the four enzymes were Leu-Gly-Gly, His-Met, Ser-Phe, and leucine amide, respectively. The mean distribution of the recovered peptide hydrolase activities against these substrates among the four enzymes I, II, III, and IV was 96.1, 1.4, 1.7, and 0.8%, respectively, for Leu-Gly-Gly; 0.6, 96.4, 2.4, and 0.6% for His-Met; 0, 0, 95.8, and 4.2% for Ser-Phe; and 20.8, 19.8, 5.6, and 53.8% for leucine amide. Ion-exchange chromatography resulted in increases in specific activity of 19-, 19-, 46-, and 3.5-fold for enzymes I, II, III, and IV, respectively. The activity of all four enzymes, but especially III and IV, were stabilized by the presence of 150 muM dithioerythritol. Activity of each of the four enzymes was decreased 79 to 100% by 1mM ethylenediaminetetraacetate, HgCl2, 1, 10-phenanthroline, or 0.5 mM p-hydroxymercuribenzoate, except that the activity of enzyme I was decreased only 15% by ethylenediaminetetraacetate. No significant activation of the partially purified enzymes occurred in the presence of 500 muM Zn++, Co++, or Mg++. The four enzymes exhibited distinct pH profiles with optima at 7.5, 7.5, 8.5, and 8.0 for enzymes I, II, III, and IV, respectively. Molecular weights of the four enzymes determined by gel filtration on Sephadex G-200 were 58,500, 74,000, 97,500, and 113,000, respectively. All four enzymes lost more than 85% of their activity after 1 hr at temperatures of 50 degrees C or higher in sodium phosphate buffer, pH 7.0. The Km values determined with the most specific substrates for each enzyme were 0.76, 0.44, 3.82, and 8.3 mM for enzymes I, II, III, and IV, respectively. Recent evidence suggests that a significant amount of some small peptides are absorbed intact and hydrolyzed by cytosol peptide hydrolases. Adequate understanding of the function and control of these intracellular enzymes requires knowledge of the characteristics and substrates specificity of individual enzymes. The study described here demonstrates the presence of at least four cytosol peptide hydrolases with distinct substrate specificities. Substrates almost exclusively hydrolyzed by each of three of the enzymes, and therefore suitable for assay of each of these enzymes in the presence of the others, have been identified. | [
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|
PMID:11150 | [Characteristics of two new mutant forms of erythrocyte glucose-6-phosphate dehydrogenase: "Kirovograd" G6PD and "Zhitomir" G6PD]. | The paper comprises the description of properties of three mutant forms of glucoso-6-phosphate dehydrogenase characterized according the WHO program. Preparations of the enzymes were isolated from erythrocytes of patients with G6PD deficiency from three unrelated to one another Ashkenasi families coming from the Ukraine and from Byelorussia. Two new variants of G6PD hitherto never described in the literature were discovered. These variants were designated as "Kirovograd" and "Zhitomir" after the towns the probands came from. The properties of purified enzymes revealed by the methods of the WHO program were as follows: the variant "Kirovograd" has a normal electrophoretic mobility in tris and TEB buffers and 98% of the normal in phosphate buffer. KM for G6P is 6,54; KM for NADP--2,19. It is characterized by a reduced thermostability and by an acute peak of activity at pH 8,5. The variant "Zhitomir" has 90-98% of the normal electrophoretic mobility in TEB buffer and 78-84% in phosphate buffer. KM for G6P is 5,4-8,3. KM for NADP is 1,4-3,1; with deoxyG6P is 50% and with deaminoNADP is 35%. It is also characterized with a reduced thermostability, while the curve of the dependence of its activity on pH has two peaks. Both variants are perfectly inactive with erythrocytes and thus should be assigned to the second group of the mutants variants of G6PD. The comparison of these variants to other variants encountered in the same national group revealed that they resemble certain quantitative variations. | [Characteristics of two new mutant forms of erythrocyte glucose-6-phosphate dehydrogenase: "Kirovograd" G6PD and "Zhitomir" G6PD]. The paper comprises the description of properties of three mutant forms of glucoso-6-phosphate dehydrogenase characterized according the WHO program. Preparations of the enzymes were isolated from erythrocytes of patients with G6PD deficiency from three unrelated to one another Ashkenasi families coming from the Ukraine and from Byelorussia. Two new variants of G6PD hitherto never described in the literature were discovered. These variants were designated as "Kirovograd" and "Zhitomir" after the towns the probands came from. The properties of purified enzymes revealed by the methods of the WHO program were as follows: the variant "Kirovograd" has a normal electrophoretic mobility in tris and TEB buffers and 98% of the normal in phosphate buffer. KM for G6P is 6,54; KM for NADP--2,19. It is characterized by a reduced thermostability and by an acute peak of activity at pH 8,5. The variant "Zhitomir" has 90-98% of the normal electrophoretic mobility in TEB buffer and 78-84% in phosphate buffer. KM for G6P is 5,4-8,3. KM for NADP is 1,4-3,1; with deoxyG6P is 50% and with deaminoNADP is 35%. It is also characterized with a reduced thermostability, while the curve of the dependence of its activity on pH has two peaks. Both variants are perfectly inactive with erythrocytes and thus should be assigned to the second group of the mutants variants of G6PD. The comparison of these variants to other variants encountered in the same national group revealed that they resemble certain quantitative variations. | [
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|
PMID:11151 | [Influence of the maternal effect on allogenic inhibition of hematopoietic stem cells]. | Bone marrow cells (0,5-10(6)) of female mice of CBA or C57BL strains were injected intravenously to lethally irradiated CBA, C57BL/6, (femaleCBA X maleC57BL/6)F1 and (femaleC57BL/6 X maleCBA)F1 mice. Spleen of recipients as assayed for colony count on the 9th day after bone marrow transplantation by the method of Till and McCullouch. Stem cells of CBA mice demonstrated failure of allogenic inhibition in (CBA X C57BL/6)F1 hybrid mice and formed the same number of colonies as in the spleen of syngenic recipients. The level of allogenic inhibition of CBA stem cells transplanted to (C57BL/6 X X CBA)F1 hybrid mice was 50%. Bone marrow cells of C57BL/6 mice formed colonies in spleen of (CBA X C57BL/6)F1 mice at least in 20 times less than in syngenic combination. In the transplantation of bone marrow from C57BL/6 mice to (C57BL/6 X CBA)F1 hybrid mice the allogenic inhibition was less pronounced (77-85%) as compared with the transfer of cells to (CBA X C57BL/6)F1 hybrid mice (95%). The sex of a recipient did not influence the number of formed colonies. The different level of allogenic inhibition of parental stem cells can not be explained by the effect of linkage with sex as the female of reciprocal hybrid mice have identical structure of sex chromosomes (X(CBA)XC57BL/6). The data obtained indicate that the maternal effect affects allogenic inhibition of stem cells in parent--F1 system. It is possible that the maternal influence may be determined by cytoplasmic factors of inheritance which affect the expressivity of recessive genes Hh, controlling the inheritance of specific haematopoietic cell antigens. | [Influence of the maternal effect on allogenic inhibition of hematopoietic stem cells]. Bone marrow cells (0,5-10(6)) of female mice of CBA or C57BL strains were injected intravenously to lethally irradiated CBA, C57BL/6, (femaleCBA X maleC57BL/6)F1 and (femaleC57BL/6 X maleCBA)F1 mice. Spleen of recipients as assayed for colony count on the 9th day after bone marrow transplantation by the method of Till and McCullouch. Stem cells of CBA mice demonstrated failure of allogenic inhibition in (CBA X C57BL/6)F1 hybrid mice and formed the same number of colonies as in the spleen of syngenic recipients. The level of allogenic inhibition of CBA stem cells transplanted to (C57BL/6 X X CBA)F1 hybrid mice was 50%. Bone marrow cells of C57BL/6 mice formed colonies in spleen of (CBA X C57BL/6)F1 mice at least in 20 times less than in syngenic combination. In the transplantation of bone marrow from C57BL/6 mice to (C57BL/6 X CBA)F1 hybrid mice the allogenic inhibition was less pronounced (77-85%) as compared with the transfer of cells to (CBA X C57BL/6)F1 hybrid mice (95%). The sex of a recipient did not influence the number of formed colonies. The different level of allogenic inhibition of parental stem cells can not be explained by the effect of linkage with sex as the female of reciprocal hybrid mice have identical structure of sex chromosomes (X(CBA)XC57BL/6). The data obtained indicate that the maternal effect affects allogenic inhibition of stem cells in parent--F1 system. It is possible that the maternal influence may be determined by cytoplasmic factors of inheritance which affect the expressivity of recessive genes Hh, controlling the inheritance of specific haematopoietic cell antigens. | [
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|
PMID:11152 | Effect of size of Morris hepatoma 5123D on gamma-glutamyltranspeptidase activity in serum and urine. | Close correlation between the size of Morris hepatoma 5123D implanted in the hind limb of rat and serum gamm-glutamyltranspeptidase activity was found. The tumour implanted in both hind legs of the rat resulted in about twofold increase of the serum enzyme activity. The growth of the hepatoma resulted also in a significant increase in the enzyme activity in urine of the tumour-bearing rats. After surgical removal of the leg with hepatoma a rapid decrease in the enzyme activity in both the studied body fluids and its subsequent renewed increase associated with formation of pulmonary metastases were observed. Partial hepatectomy and pancreatectomy were without effect on the serum gamma-glutamyltranspeptidase activity. | Effect of size of Morris hepatoma 5123D on gamma-glutamyltranspeptidase activity in serum and urine. Close correlation between the size of Morris hepatoma 5123D implanted in the hind limb of rat and serum gamm-glutamyltranspeptidase activity was found. The tumour implanted in both hind legs of the rat resulted in about twofold increase of the serum enzyme activity. The growth of the hepatoma resulted also in a significant increase in the enzyme activity in urine of the tumour-bearing rats. After surgical removal of the leg with hepatoma a rapid decrease in the enzyme activity in both the studied body fluids and its subsequent renewed increase associated with formation of pulmonary metastases were observed. Partial hepatectomy and pancreatectomy were without effect on the serum gamma-glutamyltranspeptidase activity. | [
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|
PMID:11153 | [Experimental anti-arrhythmic effects of a new beta-adrenergic receptor blocking agent, dl-l-(tert. butylamino)-3-[(2-propinyloxy)phenoxy]2-propanol hydrochloride (dl Kö 1400-Cl)]. | Antiarrhythmic property of a new adrenergic beta-blocking agent, dl-1-(tert.butylamino)-3[(2-propinyloxy) phenoxy]-2-propanol hydrochloride (Kö 1400-Cl) was studied, using 1) ouabain-induced arrhythmia in the guinea pig, 2) aconitine-induced arrhythmia in the rat, 3) arrhythmia induced by two-step ligation of coronary artery (Harris's method) in the dog and 4) halothane-adrenaline arrhythmia in the dog and was compared with those of propranolol, oxprenolol, procainamide and ajmaline. Procainamide and ajmaline produced a marked protective effect against aconitine-induced ventricular extrasystole, but were not so effective against aconitine-induced ventricular fibrillation, while oxprenolol and, to a lesser degree, propranolol were effective against the latter type of aconitine arrhythmias. Kö 1400-Cl proved to be ineffective. All the compounds tested produced a marked protective action against ouabain-arrhythmia. Whereas procainamide was most effective in abolishing the ventricular arrhythmia due to coronary-ligation even on the first postoperative day, Kö 1400-Cl and propranolol were almost ineffective on the first day. Even on the second postoperative day, the antiarrhythmic effects of these two beta-blockers were not remarkable, effective only in 2/4 animals in the case of Kö 1400-Cl and in 2/3 animals in the case of propranolol. On the contrary, all the beta-blockers tested produced a protective action against halothane-adrenaline arrhythmia at much lower doses than against coronary ligation arrhythmia. The potency ratio of Kö 1400-Cl and propranolol was 3 : 1, which paralleled with beta-blocking activity of these compounds. | [Experimental anti-arrhythmic effects of a new beta-adrenergic receptor blocking agent, dl-l-(tert. butylamino)-3-[(2-propinyloxy)phenoxy]2-propanol hydrochloride (dl Kö 1400-Cl)]. Antiarrhythmic property of a new adrenergic beta-blocking agent, dl-1-(tert.butylamino)-3[(2-propinyloxy) phenoxy]-2-propanol hydrochloride (Kö 1400-Cl) was studied, using 1) ouabain-induced arrhythmia in the guinea pig, 2) aconitine-induced arrhythmia in the rat, 3) arrhythmia induced by two-step ligation of coronary artery (Harris's method) in the dog and 4) halothane-adrenaline arrhythmia in the dog and was compared with those of propranolol, oxprenolol, procainamide and ajmaline. Procainamide and ajmaline produced a marked protective effect against aconitine-induced ventricular extrasystole, but were not so effective against aconitine-induced ventricular fibrillation, while oxprenolol and, to a lesser degree, propranolol were effective against the latter type of aconitine arrhythmias. Kö 1400-Cl proved to be ineffective. All the compounds tested produced a marked protective action against ouabain-arrhythmia. Whereas procainamide was most effective in abolishing the ventricular arrhythmia due to coronary-ligation even on the first postoperative day, Kö 1400-Cl and propranolol were almost ineffective on the first day. Even on the second postoperative day, the antiarrhythmic effects of these two beta-blockers were not remarkable, effective only in 2/4 animals in the case of Kö 1400-Cl and in 2/3 animals in the case of propranolol. On the contrary, all the beta-blockers tested produced a protective action against halothane-adrenaline arrhythmia at much lower doses than against coronary ligation arrhythmia. The potency ratio of Kö 1400-Cl and propranolol was 3 : 1, which paralleled with beta-blocking activity of these compounds. | [
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|
PMID:11154 | Exocellular proteases of Serratia marcescens and their toxicity to larvae of Galleria mellonella. | Out of 18 strains of Serratia marcescens producing exocellular proteases the strain Serratia marcescens CCEB 415 was selected according to preliminary experiments. It could be shown that the train exhibits proteolytic activity reaching up to 10 TU per 1 ml of the culture filtrate in a medium with gelatine and peptone. Two proteolytic enzyme could be demonstrated by means of specific inhibitors EDTA and diisopropyfluorophosphate: metalprotease with optimum activity at pH 7.5 and serine protease with pH optimum of 10.9. The enzymes were purified on Sephadex and DEAE cellulose columns and by means of gel electrophoresis. However, it was not possible to separate them. The optimum temperature for activity of the mixture of the two enzymes was 50degrees C, molecular weight varied around 37000 (according to gel filtration); certain kinetic characteristics of their activity were determined. Excess subtrate (casein) inhibited activity of the enzyme mixture. Toxicity of proteases expressed as LD50 units equals 78 - 10(3) TU per larva of Galleria mellonella. | Exocellular proteases of Serratia marcescens and their toxicity to larvae of Galleria mellonella. Out of 18 strains of Serratia marcescens producing exocellular proteases the strain Serratia marcescens CCEB 415 was selected according to preliminary experiments. It could be shown that the train exhibits proteolytic activity reaching up to 10 TU per 1 ml of the culture filtrate in a medium with gelatine and peptone. Two proteolytic enzyme could be demonstrated by means of specific inhibitors EDTA and diisopropyfluorophosphate: metalprotease with optimum activity at pH 7.5 and serine protease with pH optimum of 10.9. The enzymes were purified on Sephadex and DEAE cellulose columns and by means of gel electrophoresis. However, it was not possible to separate them. The optimum temperature for activity of the mixture of the two enzymes was 50degrees C, molecular weight varied around 37000 (according to gel filtration); certain kinetic characteristics of their activity were determined. Excess subtrate (casein) inhibited activity of the enzyme mixture. Toxicity of proteases expressed as LD50 units equals 78 - 10(3) TU per larva of Galleria mellonella. | [
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|
PMID:11158 | [Teratogenic damages of the male genital organs]. | 1. Malformations and functional disturbances of the male genitalia may be caused by teratogens. 2. A short review of the prenatal development points out the possible sites of action. 3. In animals some distinct teratogens produce typical malformation syndromo spermatogenetic cells. Cyproteronacetat, an antiandrogen, suppresses the development of the accessoric genital organs and produces an external feminisation. 4. In man, cryptorchidism, agenesis of the spermatic tracts, anorchia and hypospady are known as non-hereditary malformations. 5. The teratogenic etiology of some disturbances of the spermatogenesis is discussed. | [Teratogenic damages of the male genital organs]. 1. Malformations and functional disturbances of the male genitalia may be caused by teratogens. 2. A short review of the prenatal development points out the possible sites of action. 3. In animals some distinct teratogens produce typical malformation syndromo spermatogenetic cells. Cyproteronacetat, an antiandrogen, suppresses the development of the accessoric genital organs and produces an external feminisation. 4. In man, cryptorchidism, agenesis of the spermatic tracts, anorchia and hypospady are known as non-hereditary malformations. 5. The teratogenic etiology of some disturbances of the spermatogenesis is discussed. | [
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|
PMID:11162 | [Liver function in undisturbed pregnancy]. | In order to evaluate liver function during pregnancy the enzyme activities of GPT, GOT, GLDH, LDH, AP, LAP, gamma-GT and CHE were determined in 272 healthy pregnant women from the 16th week of gestation up to term. The normal range of GPT, GOT, GLDH, LDH, gamma-GT and CHE did not differ significantly from those in non-pregnant women. Increases in AP and LAP are conditioned by placental synthesis. The functional reserves of a healthy liver obviously suffice to compensate for increased demands during pregnancy. Increases in enzyme aktivities during pregnancy are not physiologic - except for increases in AP and LAP. | [Liver function in undisturbed pregnancy]. In order to evaluate liver function during pregnancy the enzyme activities of GPT, GOT, GLDH, LDH, AP, LAP, gamma-GT and CHE were determined in 272 healthy pregnant women from the 16th week of gestation up to term. The normal range of GPT, GOT, GLDH, LDH, gamma-GT and CHE did not differ significantly from those in non-pregnant women. Increases in AP and LAP are conditioned by placental synthesis. The functional reserves of a healthy liver obviously suffice to compensate for increased demands during pregnancy. Increases in enzyme aktivities during pregnancy are not physiologic - except for increases in AP and LAP. | [
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|
PMID:11163 | [Beta receptor blocking agents in psychiatry (author's transl)]. | The effects of beta receptor blocking agents in various psychiatric indications have been investigated during the last decade. The results are summarised and discussed. A final judgement of the value of beta blocking agents within the complete psychopharmacological treatment is, at present, not yet possible. They appear to have a certain therapeutic effect depending on the peripheral beta receptor blockade, at least on functional psycho- and neurovegetative cardial and circulatory disturbances as well as on anxiety states, neurotic and those caused by stress, both of these presenting primarily somatic symptoms. In other cases the results, as they now stand, present some very interesting features for further investigation. | [Beta receptor blocking agents in psychiatry (author's transl)]. The effects of beta receptor blocking agents in various psychiatric indications have been investigated during the last decade. The results are summarised and discussed. A final judgement of the value of beta blocking agents within the complete psychopharmacological treatment is, at present, not yet possible. They appear to have a certain therapeutic effect depending on the peripheral beta receptor blockade, at least on functional psycho- and neurovegetative cardial and circulatory disturbances as well as on anxiety states, neurotic and those caused by stress, both of these presenting primarily somatic symptoms. In other cases the results, as they now stand, present some very interesting features for further investigation. | [
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|
PMID:11164 | [A sensitive fluorometric determination of catechol methyltransferase activity (author's transl)]. | A method for the determination of catechol-methyltransferase activity is described, based on the measurement of fluorometric intensity of 7-hydroxy-6-methoxycoumarin (scopoletin), enzymatically produced by dihydroxycoumarin in the presence of the methyl donor S-adenosylmethionine. | [A sensitive fluorometric determination of catechol methyltransferase activity (author's transl)]. A method for the determination of catechol-methyltransferase activity is described, based on the measurement of fluorometric intensity of 7-hydroxy-6-methoxycoumarin (scopoletin), enzymatically produced by dihydroxycoumarin in the presence of the methyl donor S-adenosylmethionine. | [
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|
PMID:11165 | [Detection of multiple molecular forms of the gamma-glutamyltransferase by concanavalin A affinity chromatography (author's transl)]. | The separation of several forms of gamma-glutamyl-transferase was achieved by using concanavalin A-Sepharose columns. The enzyme of the adult liver was bound totally to the lectin, whereas only 5% of the kidney enzyme and 50% of the pancreas gamma-glutamyltransferase was adsorbed by concanavalin A. Due to a higher content of N-acetylneuraminic acid, the enzyme of the fetal liver does not show any affinity to concanavalin A. Within 8 days after birth the N-acetylneuraminic acid-rich fetal gamma-glutamyltransferase is substituted by the adult form. | [Detection of multiple molecular forms of the gamma-glutamyltransferase by concanavalin A affinity chromatography (author's transl)]. The separation of several forms of gamma-glutamyl-transferase was achieved by using concanavalin A-Sepharose columns. The enzyme of the adult liver was bound totally to the lectin, whereas only 5% of the kidney enzyme and 50% of the pancreas gamma-glutamyltransferase was adsorbed by concanavalin A. Due to a higher content of N-acetylneuraminic acid, the enzyme of the fetal liver does not show any affinity to concanavalin A. Within 8 days after birth the N-acetylneuraminic acid-rich fetal gamma-glutamyltransferase is substituted by the adult form. | [
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|
PMID:11166 | [Isoelectric focusing of complex protein mixtures in the nanogram range in microgels (author's transl)]. | A method is described for isolectric focusing of complex protein mixtures in 2, 5 or 10 mul capillaries. For one separation only 15- 50 ng of a protein mixture is needed. Isoelectric focusing is finished after 10 min, staining takes 20 min and destaining approximately 30 min. Using defined mixtures of Servalyt from different pH ranges, isoelectric focusing can be adapted to the protein sample to be fractionated. Protein peaks separated by isoelectric focusing can be electrophoretically eluted and for further analysis refractionated directly in a microgradient gel. The resolution power of microisoelectric focusing is as good as that of the wellknown macroprocedure, as is demonstrated by isoelectric focusing of the water soluble proteins from cerebellum and heart, of rat and human serum and of a human oncocytoma of the thyroid gland. | [Isoelectric focusing of complex protein mixtures in the nanogram range in microgels (author's transl)]. A method is described for isolectric focusing of complex protein mixtures in 2, 5 or 10 mul capillaries. For one separation only 15- 50 ng of a protein mixture is needed. Isoelectric focusing is finished after 10 min, staining takes 20 min and destaining approximately 30 min. Using defined mixtures of Servalyt from different pH ranges, isoelectric focusing can be adapted to the protein sample to be fractionated. Protein peaks separated by isoelectric focusing can be electrophoretically eluted and for further analysis refractionated directly in a microgradient gel. The resolution power of microisoelectric focusing is as good as that of the wellknown macroprocedure, as is demonstrated by isoelectric focusing of the water soluble proteins from cerebellum and heart, of rat and human serum and of a human oncocytoma of the thyroid gland. | [
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|
PMID:11167 | Activation of snail (Helix pomatia) nervous tissue tyrosine monooxygenase by calcium in vitro. | Addition of calcium chloride to soluble preparations of tyrosine monooxygenase from snail brain appears to produce an activation of the enzyme when assayed with subsaturating concentrations of the pteridine cofactor 6 MPH4 (2-amino-4-hydroxy-6-methyltetrahydropteridine). While some increase in the activity occurs with calcium chloride at a concentration of 0.01 mM, activation is increased by about 100% at 1mM and reaches a maximum at 5mM (144%) where it remains more or less constant up to 10mM. Barium chloride also produces an activating effect although it is much less pronounced while magnesium chloride is without effect. EGTA has no direct effect on the enzyme but antagonises the activation produced by calcium chloride. The activation of tyrosine monooxygenase by calcium is reflected in changes in the kinetic properties of the enzyme, decreasing the Km from 43 muM to 19 muM for tyrosine and from 670muM to 230muM for the pteridine cofactor. No change was observed with V values for either tyrosine or pteridine cofactor. It is suggested that calcium, which enters the nerve terminal during nerve stimulation, regulates the transmitter dopamine by activating the rate-limiting enzyme tyrosine monooxygenase. | Activation of snail (Helix pomatia) nervous tissue tyrosine monooxygenase by calcium in vitro. Addition of calcium chloride to soluble preparations of tyrosine monooxygenase from snail brain appears to produce an activation of the enzyme when assayed with subsaturating concentrations of the pteridine cofactor 6 MPH4 (2-amino-4-hydroxy-6-methyltetrahydropteridine). While some increase in the activity occurs with calcium chloride at a concentration of 0.01 mM, activation is increased by about 100% at 1mM and reaches a maximum at 5mM (144%) where it remains more or less constant up to 10mM. Barium chloride also produces an activating effect although it is much less pronounced while magnesium chloride is without effect. EGTA has no direct effect on the enzyme but antagonises the activation produced by calcium chloride. The activation of tyrosine monooxygenase by calcium is reflected in changes in the kinetic properties of the enzyme, decreasing the Km from 43 muM to 19 muM for tyrosine and from 670muM to 230muM for the pteridine cofactor. No change was observed with V values for either tyrosine or pteridine cofactor. It is suggested that calcium, which enters the nerve terminal during nerve stimulation, regulates the transmitter dopamine by activating the rate-limiting enzyme tyrosine monooxygenase. | [
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|
PMID:11168 | Extrinsic signals for monitoring the association reaction of proteins as introduced by fluorescent and non-fluorescent labels. | Two known dansyl labels (I, II) and 5-[2-(iodoacetamido)ethylamino]-1-naphthalene-sulfonic acid (III) and three new azo-dyes (IV - VI) were covalently attached to alpha-chymotrypsin and to basic pancreatic trypsin inhibitor by four different reactive groups. In order to protect the contact region of the proteins the complex of the two proteins was labeled. Advantage was taken of the fact that a group which is buried in the complex reacts about (see article) times slower than a group which is always exposed (K = dissociation equilibrium constant, [C] = concentration of the complex). The complex was dissociated at pH 3 and the labeled proteins were isolated by column chromatography. They were fully active. The dansyl label was immobilized when introduced by dansyl chloride but highly mobile when attached via the longer imidoester group (II). Changes of absorption and of fluorescence which occur when differently labeled reaction partners recombine were studied. Changes in absorption (up to 18%) were mainly due to interactions of the label of one protein with the other protein. Fluorescence changes of up to 480% could be obtained. They were interpreted in terms of a Förster type energy transfer between donor and acceptor labels and changes of absorption and quantum yield due to interactions of the labels with the proteins. The kinetic constants of complex formation are not seriously altered by the labels (Bösterling, B & Engel, J. (1976) this J. 357, 1297-1307, succeeding). It is concluded that the labeling technique may be of general value for kinetic and equilibrium studies of protein associations. | Extrinsic signals for monitoring the association reaction of proteins as introduced by fluorescent and non-fluorescent labels. Two known dansyl labels (I, II) and 5-[2-(iodoacetamido)ethylamino]-1-naphthalene-sulfonic acid (III) and three new azo-dyes (IV - VI) were covalently attached to alpha-chymotrypsin and to basic pancreatic trypsin inhibitor by four different reactive groups. In order to protect the contact region of the proteins the complex of the two proteins was labeled. Advantage was taken of the fact that a group which is buried in the complex reacts about (see article) times slower than a group which is always exposed (K = dissociation equilibrium constant, [C] = concentration of the complex). The complex was dissociated at pH 3 and the labeled proteins were isolated by column chromatography. They were fully active. The dansyl label was immobilized when introduced by dansyl chloride but highly mobile when attached via the longer imidoester group (II). Changes of absorption and of fluorescence which occur when differently labeled reaction partners recombine were studied. Changes in absorption (up to 18%) were mainly due to interactions of the label of one protein with the other protein. Fluorescence changes of up to 480% could be obtained. They were interpreted in terms of a Förster type energy transfer between donor and acceptor labels and changes of absorption and quantum yield due to interactions of the labels with the proteins. The kinetic constants of complex formation are not seriously altered by the labels (Bösterling, B & Engel, J. (1976) this J. 357, 1297-1307, succeeding). It is concluded that the labeling technique may be of general value for kinetic and equilibrium studies of protein associations. | [
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|
PMID:11169 | Influence of various fluorescent and non-fluorescent labels on the kinetics of the complex formation of alpha-chymotrypsin with basic pancreatic trypsin inhibitor (Kunitz). | The association of alpha-chymotrypsin with basic pancreatic trypsin inhibitor was studied using extrinsic signals produced by fluorescent and nonfluorescent labels. The reactive dyes were covalently bound to the proteins in the complexed state, in which the binding region was protected. The signals were sufficiently large to measure the complex formation at protein concentrations of 10(-9)M by fluorescence and down to 10(-6)M by absorption. Therefore, the association and dissociation could be followed over a broad range of concentration. Good correspondence was observed between data which were obtained with different labels and with published values for the unlabeled proteins. Existing differences could be explained by different buffer conditions used by the different authors. Also the pH dependence of the dissociation rate constants was essentially unaltered by the introduction of the labels. The large signals allowed a direct measurement of the equilibrium constants of dissociation, even at high pH, at which they are in the range of 10(-8)M. The experimentally determined binding constants were in agreement with those calculated from the rate constants. The temperature dependence of the binding constants revealed a small positive and pH-dependent enthalpy change [deltaHo = 4.0 kcal/mol (16.7 kJ) at H 7.0[. The results prove that the labeling can be performed in such a way that the equilibrium and kinetic parameters of the system studied are not significantly influenced. | Influence of various fluorescent and non-fluorescent labels on the kinetics of the complex formation of alpha-chymotrypsin with basic pancreatic trypsin inhibitor (Kunitz). The association of alpha-chymotrypsin with basic pancreatic trypsin inhibitor was studied using extrinsic signals produced by fluorescent and nonfluorescent labels. The reactive dyes were covalently bound to the proteins in the complexed state, in which the binding region was protected. The signals were sufficiently large to measure the complex formation at protein concentrations of 10(-9)M by fluorescence and down to 10(-6)M by absorption. Therefore, the association and dissociation could be followed over a broad range of concentration. Good correspondence was observed between data which were obtained with different labels and with published values for the unlabeled proteins. Existing differences could be explained by different buffer conditions used by the different authors. Also the pH dependence of the dissociation rate constants was essentially unaltered by the introduction of the labels. The large signals allowed a direct measurement of the equilibrium constants of dissociation, even at high pH, at which they are in the range of 10(-8)M. The experimentally determined binding constants were in agreement with those calculated from the rate constants. The temperature dependence of the binding constants revealed a small positive and pH-dependent enthalpy change [deltaHo = 4.0 kcal/mol (16.7 kJ) at H 7.0[. The results prove that the labeling can be performed in such a way that the equilibrium and kinetic parameters of the system studied are not significantly influenced. | [
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|
PMID:11177 | Rat liver cells in culture: effect of storage, long-term culture, and transformation on some enzyme levels. | Aryl hydrocarbon hydroxylase (AHH) and tyrosine aminotransferase (TAT) activities were determined in rat liver cell lines after frozen storage, long-term culture, and transformation in vitro. Levels of AHH activity after 17 months in frozen storage were comparable to levels prior to freezing. During long-term culture the AHH levels of the cell lines tended to decrease. Transformed lines had variable levels of AHH activity. Cell lines retained measurable TAT activity following long-term culture and frozen storage. TAT activity of transformed cells was comparable to that of normal lines. Prolonged frozen storage did not induce transformation up to one year. | Rat liver cells in culture: effect of storage, long-term culture, and transformation on some enzyme levels. Aryl hydrocarbon hydroxylase (AHH) and tyrosine aminotransferase (TAT) activities were determined in rat liver cell lines after frozen storage, long-term culture, and transformation in vitro. Levels of AHH activity after 17 months in frozen storage were comparable to levels prior to freezing. During long-term culture the AHH levels of the cell lines tended to decrease. Transformed lines had variable levels of AHH activity. Cell lines retained measurable TAT activity following long-term culture and frozen storage. TAT activity of transformed cells was comparable to that of normal lines. Prolonged frozen storage did not induce transformation up to one year. | [
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|
PMID:11178 | Synthesis of alpha subunit of human chorionic gonadotrophin by presumptive HeLa cells. | Several cell lines, originally thought to be derived from a human placenta at term but possibly HeLa-contaminated, have been studied. These cells secrete a protein indistinguishable immunochemically from the alpha subunit of chorionic gonadotropin but not the beta subunit of chorionic gonadotropin or placental lactogen. Complete chorionic gonadotropin was detected but amounted to less than 1% of the level of the alpha subunit. The cells also produce an alkaline phosphatase similar to placental alkaline phosphatase in immunochemical, gel-electrophoretic, and heat-denaturation properties. They induce tumor growth when inoculated into nude mice. These cells are aneuploid and have a model chromosome number of 66. The common HeLa karyologic markers, designated 1, 2, and 3, and A-type glucose-6-phosphate dehydrogenase are present in these cells. HeLa cells have not previously been shown to secrete the alpha subunit of hCG. | Synthesis of alpha subunit of human chorionic gonadotrophin by presumptive HeLa cells. Several cell lines, originally thought to be derived from a human placenta at term but possibly HeLa-contaminated, have been studied. These cells secrete a protein indistinguishable immunochemically from the alpha subunit of chorionic gonadotropin but not the beta subunit of chorionic gonadotropin or placental lactogen. Complete chorionic gonadotropin was detected but amounted to less than 1% of the level of the alpha subunit. The cells also produce an alkaline phosphatase similar to placental alkaline phosphatase in immunochemical, gel-electrophoretic, and heat-denaturation properties. They induce tumor growth when inoculated into nude mice. These cells are aneuploid and have a model chromosome number of 66. The common HeLa karyologic markers, designated 1, 2, and 3, and A-type glucose-6-phosphate dehydrogenase are present in these cells. HeLa cells have not previously been shown to secrete the alpha subunit of hCG. | [
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PMID:11179 | [Approach to a practical method for screening and identifying microorganism genera from urine (author's transl)]. | In this study the author reported upon a practical new system for screening and identifying the microbial agents causing urinary tract infections. This system is composed of a combination of 3 screening procedures (pH-value + nitrite-test + catalase-test) and 8 selective culture media for the purpose of genus identification within 24 hours (Uripret-G). A total of 130 cultures was investigated. The employed microorganisms were mainly recovered from urine samples. They included the following species: Candida albicans, Citrobacter freundii, Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus inconstans, Proteus mirabilis, Proteus morganii, Proteus rettgeri, Proteus vulgaris, Serratia liquefaciens, Serratia marcescens, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus faecalis and Streptococcus faecium. Employing coded cultures not only monoinfections but also multiinfections in urine samples were simulated. Under the circumstances of investigation it was possible with the help of the new system to reidentify the genera of all but two of the 130 employed microorganisms. | [Approach to a practical method for screening and identifying microorganism genera from urine (author's transl)]. In this study the author reported upon a practical new system for screening and identifying the microbial agents causing urinary tract infections. This system is composed of a combination of 3 screening procedures (pH-value + nitrite-test + catalase-test) and 8 selective culture media for the purpose of genus identification within 24 hours (Uripret-G). A total of 130 cultures was investigated. The employed microorganisms were mainly recovered from urine samples. They included the following species: Candida albicans, Citrobacter freundii, Enterobacter aerogenes, Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus inconstans, Proteus mirabilis, Proteus morganii, Proteus rettgeri, Proteus vulgaris, Serratia liquefaciens, Serratia marcescens, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus faecalis and Streptococcus faecium. Employing coded cultures not only monoinfections but also multiinfections in urine samples were simulated. Under the circumstances of investigation it was possible with the help of the new system to reidentify the genera of all but two of the 130 employed microorganisms. | [
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|
PMID:11180 | Slow reacting substance as a preformed mediator from human lung. | Homogenates from human lung contained a preformed slow reacting substance (pSRS). The pattern of contraction on the guinea-pig ileum by pSRS was indistinguishable from that of SRS-A. The activity of pSRS could not be attributed to the presence of K+, Na+, Ca2+ and Mg2+ ions, or any prostaglandin including PGF2 or its 15-oxo derivative. As with SRS-A, pSRS could be absorbed onto Amberlite XAD-2 and silicic acid. Both were eluted from the former with 80 per cent ethanol and from the latter with a mixture of ethanol, ammonia and water. Both pSRS and SRS-A were resistant to the action of NaOH whereas their activities were destroyed by boiling in HCl. Arylsulphatase II B destroyed the activities of both pSRS and SRS-A. An antagonist of SRS-A, FPL55712, inhibited the action of pSRS at comparable concentrations to that of SRS-A. These experiments suggest that pSRS and SRS-A are identical. Thus SRS joins histamine and ECF-A as a preformed mediator. Although SRS was present in a preformed state the amount of material extractable was more than doubled by the anaphylactic reaction. The extraction of slow reacting substance from human lung without apparent requirement for antigen or antibody points to a possible role of this mediator in inflammatory reactions evoked by mechanisms independent of IgE and other tissue-sensitizing antibodies. | Slow reacting substance as a preformed mediator from human lung. Homogenates from human lung contained a preformed slow reacting substance (pSRS). The pattern of contraction on the guinea-pig ileum by pSRS was indistinguishable from that of SRS-A. The activity of pSRS could not be attributed to the presence of K+, Na+, Ca2+ and Mg2+ ions, or any prostaglandin including PGF2 or its 15-oxo derivative. As with SRS-A, pSRS could be absorbed onto Amberlite XAD-2 and silicic acid. Both were eluted from the former with 80 per cent ethanol and from the latter with a mixture of ethanol, ammonia and water. Both pSRS and SRS-A were resistant to the action of NaOH whereas their activities were destroyed by boiling in HCl. Arylsulphatase II B destroyed the activities of both pSRS and SRS-A. An antagonist of SRS-A, FPL55712, inhibited the action of pSRS at comparable concentrations to that of SRS-A. These experiments suggest that pSRS and SRS-A are identical. Thus SRS joins histamine and ECF-A as a preformed mediator. Although SRS was present in a preformed state the amount of material extractable was more than doubled by the anaphylactic reaction. The extraction of slow reacting substance from human lung without apparent requirement for antigen or antibody points to a possible role of this mediator in inflammatory reactions evoked by mechanisms independent of IgE and other tissue-sensitizing antibodies. | [
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|
PMID:11181 | Analysis of immunosuppression generated by the graft-versus-host reaction. II. Characterization of the suppression cell and its mechanism of action. | Spleen cells from (CBA X C57/BL) F1 mice undergoing graft-versus-host (GVH) reaction induced by injection of parental cells 7-14 days previously are capable of suppressing an immune response by normal or primed F1 spleen cells to chicken erythrocytes and levan in vivo and sheep erythrocytes in vitro. The cells in these GVH spleens which were responsible for the suppression were sensitive to treatment with anti-0 serum, resistant to 900 rad irradiation in vivo and not retained by anti-immunoglobulin columns. Suppressor activity in vitro was present only in the non-adherent fraction of these GVH cell suspensions. Furthermore, the T-cell fraction, purified by affinity chromatography, suppressed the in vitro response of macrophage-depleted normal F1 cells to DNP-levan. Collectively, these observations imply that suppressor T cells generated by GVH reaction can affect B-cell functions directly without intermediary macrophage participation. Spleen cells from (CBA X C57/BL) F1 mice undergoing GVH reaction induced by C57/BL cells were depleted of their F1 content by treatment with anti-CBA alloantiserum. The suppressive activity of the residual donor component was still expressed against other F1 cells (AKR X C57/BL) which were H-2 compatible with the original host, but not against H-2-incompatible cells (DBA/1 X C57/BL) F1. However, the latter were suppressed in the presence of (CBA X C57/BL) F1 cells. Thus, interaction of donor T cells with F1 target cells containing those H-2 antigens towards which they are sensitized is mandatory for the subsequent manifestation of immunosuppressive activity. GVH cells suppressed the response of primed F1 cells in double Marbrook chambers when the two populations were separated were by a cell-impermeable membrane, provided the GVH suspension contained F1 cells to which donor T cells were sensitized. This suggests that soluble factors are involved in the mechanism of GVH-induced immunosuppression. | Analysis of immunosuppression generated by the graft-versus-host reaction. II. Characterization of the suppression cell and its mechanism of action. Spleen cells from (CBA X C57/BL) F1 mice undergoing graft-versus-host (GVH) reaction induced by injection of parental cells 7-14 days previously are capable of suppressing an immune response by normal or primed F1 spleen cells to chicken erythrocytes and levan in vivo and sheep erythrocytes in vitro. The cells in these GVH spleens which were responsible for the suppression were sensitive to treatment with anti-0 serum, resistant to 900 rad irradiation in vivo and not retained by anti-immunoglobulin columns. Suppressor activity in vitro was present only in the non-adherent fraction of these GVH cell suspensions. Furthermore, the T-cell fraction, purified by affinity chromatography, suppressed the in vitro response of macrophage-depleted normal F1 cells to DNP-levan. Collectively, these observations imply that suppressor T cells generated by GVH reaction can affect B-cell functions directly without intermediary macrophage participation. Spleen cells from (CBA X C57/BL) F1 mice undergoing GVH reaction induced by C57/BL cells were depleted of their F1 content by treatment with anti-CBA alloantiserum. The suppressive activity of the residual donor component was still expressed against other F1 cells (AKR X C57/BL) which were H-2 compatible with the original host, but not against H-2-incompatible cells (DBA/1 X C57/BL) F1. However, the latter were suppressed in the presence of (CBA X C57/BL) F1 cells. Thus, interaction of donor T cells with F1 target cells containing those H-2 antigens towards which they are sensitized is mandatory for the subsequent manifestation of immunosuppressive activity. GVH cells suppressed the response of primed F1 cells in double Marbrook chambers when the two populations were separated were by a cell-impermeable membrane, provided the GVH suspension contained F1 cells to which donor T cells were sensitized. This suggests that soluble factors are involved in the mechanism of GVH-induced immunosuppression. | [
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|
PMID:11187 | Large-scale purification and characterization of the exotoxin of Pseudomonas aeruginosa. | The exotoxin (PE) of Pseudomonas aeruginosa was purified from 50-liter cultures by a simple three-step procedure, yielding 135 mg of essentially homogeneous protein. In Ouchterlony gel diffusion, PE produces a single line which does not interact with a diphtheria toxin-antitoxin precipitin line. The protein has a molecular weight of 66,000, an isoelectric point of 5.1, N-terminal arginine, and four disulfide bridges. The amino acid composition shows no apparent similarity to that of diphtheria toxin. The median lethal dose of this PE preparation in mice weighing 20 g is 0.1 mug. The median lethal dose in 350-g rats is 20 mug. The cytotoxicity of PE for mouse L929 fibroblasts is completely neutralized by small amounts of specific pony antitoxin. The exotoxin possesses adenosine diphosphate-ribosylation activity. Both cytotoxic and adenosine diphosphate-ribosylation activities are shown to be properties of the intact 66,000-dalton protein. | Large-scale purification and characterization of the exotoxin of Pseudomonas aeruginosa. The exotoxin (PE) of Pseudomonas aeruginosa was purified from 50-liter cultures by a simple three-step procedure, yielding 135 mg of essentially homogeneous protein. In Ouchterlony gel diffusion, PE produces a single line which does not interact with a diphtheria toxin-antitoxin precipitin line. The protein has a molecular weight of 66,000, an isoelectric point of 5.1, N-terminal arginine, and four disulfide bridges. The amino acid composition shows no apparent similarity to that of diphtheria toxin. The median lethal dose of this PE preparation in mice weighing 20 g is 0.1 mug. The median lethal dose in 350-g rats is 20 mug. The cytotoxicity of PE for mouse L929 fibroblasts is completely neutralized by small amounts of specific pony antitoxin. The exotoxin possesses adenosine diphosphate-ribosylation activity. Both cytotoxic and adenosine diphosphate-ribosylation activities are shown to be properties of the intact 66,000-dalton protein. | [
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|
PMID:11188 | Serum bactericidal activity in the horseshoe crab, Limulus polyphemus. | Serum from the horseshoe crab, Limulus polyphemus, was examined for bactericidal activity against five species of bacteria. Greatest activity was found against Pseudomonas putida and Flavobacterium sp.; with the former, serum dilutions as high as 1:20 were capable of reducing viable counts by 50% within 2 h. Bactericidal activity of a significantly lesser magnitude was demonstrated against Serratia marcesencs and Salmonella minnesota. No killing was seen when the lobster pathogen Aerococcus viridans (formerly Gaffkya homari) was used. The cidal activity against Flavobacterium sp. remained relatively consistent for 6 months. | Serum bactericidal activity in the horseshoe crab, Limulus polyphemus. Serum from the horseshoe crab, Limulus polyphemus, was examined for bactericidal activity against five species of bacteria. Greatest activity was found against Pseudomonas putida and Flavobacterium sp.; with the former, serum dilutions as high as 1:20 were capable of reducing viable counts by 50% within 2 h. Bactericidal activity of a significantly lesser magnitude was demonstrated against Serratia marcesencs and Salmonella minnesota. No killing was seen when the lobster pathogen Aerococcus viridans (formerly Gaffkya homari) was used. The cidal activity against Flavobacterium sp. remained relatively consistent for 6 months. | [
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|
PMID:11189 | Immunity to Mycobacterium leprae infections in mice stimulated by M. leprae, BCG, and graft-versus-host reactions. | Infections of mice with Mycobacterium leprae in one rear foot pad immunized them against a second infection in the other rear foot pad. Purified bacilli harvested from the first infection also produced immuniy when injection into the foot pads of previously uninfected mice. Injections of BCG afforded similar protection, but had no adjuvant effect on M. leprae. M. duvali, a cultivable mycobacterium that is reported to be more closely related antigenically to M. leprae than BCG is, provided much less protection against M. leprae challenge than BCG did. Moreover, when M. duvali was mixed with BCG, it was not any more effective than BCG alone. Graft-versus-host reactions, induced by injections of parental spleen cells into F1 hybrids, provided no protection against M. tuberculosis and M. marinum challenge. They gave moderate protection against M. leprae in one experiment but not in another with a different schedule. Allogenic spleen cells had a protective effect when injected locally into the infected foot pad. The effect produced by these injections of spleen cells was a delay in the appearance of bacterial growth; however, there was no decrease in the rate of logarithmic growth when it did appear and no reduction in the eventual plateau level. | Immunity to Mycobacterium leprae infections in mice stimulated by M. leprae, BCG, and graft-versus-host reactions. Infections of mice with Mycobacterium leprae in one rear foot pad immunized them against a second infection in the other rear foot pad. Purified bacilli harvested from the first infection also produced immuniy when injection into the foot pads of previously uninfected mice. Injections of BCG afforded similar protection, but had no adjuvant effect on M. leprae. M. duvali, a cultivable mycobacterium that is reported to be more closely related antigenically to M. leprae than BCG is, provided much less protection against M. leprae challenge than BCG did. Moreover, when M. duvali was mixed with BCG, it was not any more effective than BCG alone. Graft-versus-host reactions, induced by injections of parental spleen cells into F1 hybrids, provided no protection against M. tuberculosis and M. marinum challenge. They gave moderate protection against M. leprae in one experiment but not in another with a different schedule. Allogenic spleen cells had a protective effect when injected locally into the infected foot pad. The effect produced by these injections of spleen cells was a delay in the appearance of bacterial growth; however, there was no decrease in the rate of logarithmic growth when it did appear and no reduction in the eventual plateau level. | [
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|
PMID:11191 | The distribution of hepatitis B surface antigen in Africa and the tropics: report of a population study in Nigeria. | This preliminary study was designed to examine the distribution of hepatitis B surface antigen (HBsAg) in two contrasting groups in an urban area situated in the tropical forest belt. The sample from the traditional area represents a population of low socio-economic status, living in the central slum areas of the city, and the sample from the peripheral area represents a population of high socio-economic status living in clean modern estates. The prevalence rate of HBsAg by complement fixation (CF) was 12-6 per cent in both areas. There was no statistically significant difference between the two groups with respect to prevalence of the antigen. When both groups were combined, no significant relationship was found between the presence of the antigen and sex, age, marital status, level of education, occupation, income, and a presumed exposure to the antigen from injections, dental treatment, blood tests, surgical operations, blood donations, tribal, (medicinal), tattoo and cosmetic marking, insanitary disposal of faeces, doubtful sources of water supply, and exposure to mosquitoes. No association with genotype was found. | The distribution of hepatitis B surface antigen in Africa and the tropics: report of a population study in Nigeria. This preliminary study was designed to examine the distribution of hepatitis B surface antigen (HBsAg) in two contrasting groups in an urban area situated in the tropical forest belt. The sample from the traditional area represents a population of low socio-economic status, living in the central slum areas of the city, and the sample from the peripheral area represents a population of high socio-economic status living in clean modern estates. The prevalence rate of HBsAg by complement fixation (CF) was 12-6 per cent in both areas. There was no statistically significant difference between the two groups with respect to prevalence of the antigen. When both groups were combined, no significant relationship was found between the presence of the antigen and sex, age, marital status, level of education, occupation, income, and a presumed exposure to the antigen from injections, dental treatment, blood tests, surgical operations, blood donations, tribal, (medicinal), tattoo and cosmetic marking, insanitary disposal of faeces, doubtful sources of water supply, and exposure to mosquitoes. No association with genotype was found. | [
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|
PMID:11192 | Thermal fragmentation of Escherichia coli beta-galactosidase. Isolation and characterization of an alpha-complementing and two non-complementing polypeptide fractions. | Carboxymethylated Escherichia coli beta-galactosidase EC 3.2.1.23 could be broken to polypeptides of fairly uniform size (average molecular weight about 22,000 daltons) by heating for less than or equal to 8 h at 100 degrees C and pH 7.5 IN 8 M-urea. Using phosphocellulose chromatography in NaCl-urea gradients, the resulting polypeptide mixture could be resolved in three fractions essentially homogeneous by disc gel electrophoresis in urea at several pH values, and by isoelectric focusing. One of these fractions was active as alpha-donor in in vitro complementation of beta-galactosidase activity with Escherichia coli mutant M15; this activity was largely retained after CNBr cleavage. All three fractions carried arginine as carboxyl-terminal amino acid. No significant amount of any specific amino could be detected in NH2-terminal position. | Thermal fragmentation of Escherichia coli beta-galactosidase. Isolation and characterization of an alpha-complementing and two non-complementing polypeptide fractions. Carboxymethylated Escherichia coli beta-galactosidase EC 3.2.1.23 could be broken to polypeptides of fairly uniform size (average molecular weight about 22,000 daltons) by heating for less than or equal to 8 h at 100 degrees C and pH 7.5 IN 8 M-urea. Using phosphocellulose chromatography in NaCl-urea gradients, the resulting polypeptide mixture could be resolved in three fractions essentially homogeneous by disc gel electrophoresis in urea at several pH values, and by isoelectric focusing. One of these fractions was active as alpha-donor in in vitro complementation of beta-galactosidase activity with Escherichia coli mutant M15; this activity was largely retained after CNBr cleavage. All three fractions carried arginine as carboxyl-terminal amino acid. No significant amount of any specific amino could be detected in NH2-terminal position. | [
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|
PMID:11193 | The effects of hallucinogens on blind monkeys. | Two blind monkeys were studied with an observational profile that was previously shown to distinguish the effects of hallucinogens from those of other classes of drugs. Lysergic acid diethylamide and dimethyltryptamine could be distinguished from saline, chlorpromazine, d-amphetamine sulfate, and bromo-lysergic acid diethylamide by the increased frequency of spasms, stereotypy, bump, and tracking. The hallucinogens also produced dramatic increases in exploration and related behaviors normally seen only in response to real visual or auditory stimuli. These behaviors are discussed in terms of their similarity to behaviors observed with sighted monkeys in light and dark environments. | The effects of hallucinogens on blind monkeys. Two blind monkeys were studied with an observational profile that was previously shown to distinguish the effects of hallucinogens from those of other classes of drugs. Lysergic acid diethylamide and dimethyltryptamine could be distinguished from saline, chlorpromazine, d-amphetamine sulfate, and bromo-lysergic acid diethylamide by the increased frequency of spasms, stereotypy, bump, and tracking. The hallucinogens also produced dramatic increases in exploration and related behaviors normally seen only in response to real visual or auditory stimuli. These behaviors are discussed in terms of their similarity to behaviors observed with sighted monkeys in light and dark environments. | [
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|
PMID:11194 | A review of psychotropic medications and the glaucomas. | Four general groups of psychotropic drugs are examined with respect to possible adverse effects upon intraocular pressure. After a brief discussion of the various mechanisms of the glaucomas, both general and specific information is provided and discussed concerning antipsychotic, antidepressant, antiparkinsonian and antianxiety preparations of a variety of chemical structures and utilities. The consensus of the authors is that with certain basic safeguards virtually all of the medications studied are acceptably safe to prescribe even in patients with diagnosed glaucoma. | A review of psychotropic medications and the glaucomas. Four general groups of psychotropic drugs are examined with respect to possible adverse effects upon intraocular pressure. After a brief discussion of the various mechanisms of the glaucomas, both general and specific information is provided and discussed concerning antipsychotic, antidepressant, antiparkinsonian and antianxiety preparations of a variety of chemical structures and utilities. The consensus of the authors is that with certain basic safeguards virtually all of the medications studied are acceptably safe to prescribe even in patients with diagnosed glaucoma. | [
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|
PMID:11197 | [Prolactin in male reproduction]. | Prolactin (PRL), a peptide hormon from the hypophysis, becomes more interesting, since it can be determined by radioimmunassays. The release of prolactin is controlled by two not yet identified factors, the prolactin-releasing-factor and the prolactin-inhibiting-factor. The latter predominantes. Many pharmacological substances can alter the release. The normal serum levels in the man are 6-13 ng/ml. Prolactin affects many organs, f.e. kidney, mamma, ovary, testis, hepar and skin. For clinical tests the raise in the serum levels after TSH or chlorpromacin and the drop after application of 2-brom-alpha-ergocryptin is used. Increased serum levels are found in patients with prolactin-producing tumors. In the male this is followed by disturbances of the sexual potency. Relations between prolactin and male infertility of gynecomastia are not yet known. | [Prolactin in male reproduction]. Prolactin (PRL), a peptide hormon from the hypophysis, becomes more interesting, since it can be determined by radioimmunassays. The release of prolactin is controlled by two not yet identified factors, the prolactin-releasing-factor and the prolactin-inhibiting-factor. The latter predominantes. Many pharmacological substances can alter the release. The normal serum levels in the man are 6-13 ng/ml. Prolactin affects many organs, f.e. kidney, mamma, ovary, testis, hepar and skin. For clinical tests the raise in the serum levels after TSH or chlorpromacin and the drop after application of 2-brom-alpha-ergocryptin is used. Increased serum levels are found in patients with prolactin-producing tumors. In the male this is followed by disturbances of the sexual potency. Relations between prolactin and male infertility of gynecomastia are not yet known. | [
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|
PMID:11198 | Rapid microspectrofluorometric studies in EL2 cells following intracellular accumulation of dibenzocarbazoles. | Microspectrofluorometric observations were carried out in EL2 ascites cancer cells and dibenzo(a,e)fluoranthene (diB(a,e)F)-grown EL2 cells, following treatment (5 min) with three dibenzocarbazoles (1,2,7,8; 1,2,5,6 and 3,4,5,6). After microinjection of glucose-6-P leading to reduction of NAD(P), a sequence of difference spectra (after substrate minus before) is recorded. In dibenzocarbazole-untreated cells, maximum (NAD(P) reduction (emission maximum at 465-475 nm) is attained within 5 s, followed by a gradual return to initial fluorescence within 20 to 200 s (faster in the diB(a,e)F-grown). In dibenzocarbazole-treated cells there is a rather regular increase in the intensity of the difference spectrum up to approximately 300-500 s. Initially the increase is more predominant in the region around 460-470 nm, but it gains later prominence in the shorter wavelength region (420-430 nm) characteristic of the hydrocarbon (higher and steadier increase in the 3,4,5,6, dibenzocarbazole-treated diB(a,e)F-grown). Subsequently there is a gradual decrease of fluorescence which may or may or not return to initial level. The observed increase spectra require evaluation in terms of possible components (e.g. a mixture of NAD(P)H and hydrocarbon, binding changes, succession of fluorescent metabolites). | Rapid microspectrofluorometric studies in EL2 cells following intracellular accumulation of dibenzocarbazoles. Microspectrofluorometric observations were carried out in EL2 ascites cancer cells and dibenzo(a,e)fluoranthene (diB(a,e)F)-grown EL2 cells, following treatment (5 min) with three dibenzocarbazoles (1,2,7,8; 1,2,5,6 and 3,4,5,6). After microinjection of glucose-6-P leading to reduction of NAD(P), a sequence of difference spectra (after substrate minus before) is recorded. In dibenzocarbazole-untreated cells, maximum (NAD(P) reduction (emission maximum at 465-475 nm) is attained within 5 s, followed by a gradual return to initial fluorescence within 20 to 200 s (faster in the diB(a,e)F-grown). In dibenzocarbazole-treated cells there is a rather regular increase in the intensity of the difference spectrum up to approximately 300-500 s. Initially the increase is more predominant in the region around 460-470 nm, but it gains later prominence in the shorter wavelength region (420-430 nm) characteristic of the hydrocarbon (higher and steadier increase in the 3,4,5,6, dibenzocarbazole-treated diB(a,e)F-grown). Subsequently there is a gradual decrease of fluorescence which may or may or not return to initial level. The observed increase spectra require evaluation in terms of possible components (e.g. a mixture of NAD(P)H and hydrocarbon, binding changes, succession of fluorescent metabolites). | [
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|
PMID:11196 | Full-time course studies of bovine liver glutamate dehydrogenase. Simulation of inhibition by pyridoxal-5'-phosphate. | Inhibition of bovine liver glutamate dehydrogenase by pyridoxal-5'-phosphate was studied by measuring the full time course of the oxidation of NADPH. Progress curves were determined before and after incubation of the enzyme with PLP in the presence of saturating concentrations of alpha-ketoglutarate and ammonium ion, at pH 7.4 and 25 degrees C. The data were fitted to the integrated Michaelis-Menten equation and an inhibition model derived. According to the model, PLP inhibits the enzyme non-competitively, by reversible formation of the complexes E--PLP and E--PLP--NADPH; the oxidation of NADPH is also inhibited by NADP+. After incubation with PLP, the dissociation constants of E--NADPH and E--NADP+ (Km and Kp) show a very definite decrease, while the maximum rate of oxidation (Vm) is increased. The inhibition constants for PLP were also computed. | Full-time course studies of bovine liver glutamate dehydrogenase. Simulation of inhibition by pyridoxal-5'-phosphate. Inhibition of bovine liver glutamate dehydrogenase by pyridoxal-5'-phosphate was studied by measuring the full time course of the oxidation of NADPH. Progress curves were determined before and after incubation of the enzyme with PLP in the presence of saturating concentrations of alpha-ketoglutarate and ammonium ion, at pH 7.4 and 25 degrees C. The data were fitted to the integrated Michaelis-Menten equation and an inhibition model derived. According to the model, PLP inhibits the enzyme non-competitively, by reversible formation of the complexes E--PLP and E--PLP--NADPH; the oxidation of NADPH is also inhibited by NADP+. After incubation with PLP, the dissociation constants of E--NADPH and E--NADP+ (Km and Kp) show a very definite decrease, while the maximum rate of oxidation (Vm) is increased. The inhibition constants for PLP were also computed. | [
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|
PMID:11199 | Adenosine triphosphate hydrolysis in rat dental tissues. A histochemical study of ion dependencies. | The effect of EDTA-decalcification, reactivating and activating procedures on the hydrolysis of ATP was studied histochemically in developing dental tissues in the rat. The incubation media contained lead citrate at alkaline pH and lead nitrate at neutral pH, and the results with ATP as substrate were compared with those obtained with beta-glycerophosphate. The ion dependency of ATP hydrolysis could only be ascertained in decalcified sections. As in earlier studies on the hydrolysis of beta-glycerophosphate in dental tissues, this hydrolysis could readily be reactivated through preincubation of the sections in a series of 0.1 M solutions of divalent cations; Zn2+ being the most efficient. This treatment was now found also to give rise to an ATP hydrolysis, which occurred without the need for activating ions in the incubation medium. This ATP hydrolysis should thus be described as nonspecific and, in terms of ion dependency, as due to a metalloenzyme, i.e. alkaline phosphatase. Activating ion dependent ATP hydrolysis in the dental tissues was found in the blood vessels and in the apical part of the secretory ameloblasts. The former was activated by Mg2+, Ca2+ and Mn2+, and the latter by Ca2+ and--almost specifically--by Sr2+. Preincubation with Zn2+ always inhibited the ion dependant ATP hydrolysis in the dental tissues. | Adenosine triphosphate hydrolysis in rat dental tissues. A histochemical study of ion dependencies. The effect of EDTA-decalcification, reactivating and activating procedures on the hydrolysis of ATP was studied histochemically in developing dental tissues in the rat. The incubation media contained lead citrate at alkaline pH and lead nitrate at neutral pH, and the results with ATP as substrate were compared with those obtained with beta-glycerophosphate. The ion dependency of ATP hydrolysis could only be ascertained in decalcified sections. As in earlier studies on the hydrolysis of beta-glycerophosphate in dental tissues, this hydrolysis could readily be reactivated through preincubation of the sections in a series of 0.1 M solutions of divalent cations; Zn2+ being the most efficient. This treatment was now found also to give rise to an ATP hydrolysis, which occurred without the need for activating ions in the incubation medium. This ATP hydrolysis should thus be described as nonspecific and, in terms of ion dependency, as due to a metalloenzyme, i.e. alkaline phosphatase. Activating ion dependent ATP hydrolysis in the dental tissues was found in the blood vessels and in the apical part of the secretory ameloblasts. The former was activated by Mg2+, Ca2+ and Mn2+, and the latter by Ca2+ and--almost specifically--by Sr2+. Preincubation with Zn2+ always inhibited the ion dependant ATP hydrolysis in the dental tissues. | [
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|
PMID:11200 | [An improved histofluorescence procedure for freeze-dried paraffin-embedded tissue based on combined formaldehyde-glyoxylic acid perfusion with high magnesium content and acid pH]. | A technique is described for highly sensitive and precise visualization of central catecholamine systems in paraffin sections of freeze-dried tissue. The procedure is based on perfusion of the animal with a solution containing formaldehyde and/or glyoxylic acid, in the presence of a very high magnesium content (40 g MgSO4/150 ml solution) and acid pH. The perfused tissue is rapidly frozen, freeze-dried, treated with formaldehyde vapours (at +80 degrees C for 1 h), embedded in parffin in vacuo, and finally sectioned. The present technique has a sensitivity for the dopamine- and noradrenaline-containing systems that is comparable with that of the glyoxylic acid-Vibratome technique, which utilizes fresh, glyoxylic acid-perfused tissue. Thus, the preterminal axon pathways become fluorescent throughout their full extent and the several new terminal systems, discovered with the glyoxylic acid-Vibratome method, are well demonstrable. The method is also highly useful for the study of the cell bodies and their dendritic processes. The catacholamine fibre systems are visualized without any signs of diffusion and with a richness in detail. In animals pretreated with L-tryptophan and MAO-inhibitor the technique is also useful for studies on central indolamine-containing systems. | [An improved histofluorescence procedure for freeze-dried paraffin-embedded tissue based on combined formaldehyde-glyoxylic acid perfusion with high magnesium content and acid pH]. A technique is described for highly sensitive and precise visualization of central catecholamine systems in paraffin sections of freeze-dried tissue. The procedure is based on perfusion of the animal with a solution containing formaldehyde and/or glyoxylic acid, in the presence of a very high magnesium content (40 g MgSO4/150 ml solution) and acid pH. The perfused tissue is rapidly frozen, freeze-dried, treated with formaldehyde vapours (at +80 degrees C for 1 h), embedded in parffin in vacuo, and finally sectioned. The present technique has a sensitivity for the dopamine- and noradrenaline-containing systems that is comparable with that of the glyoxylic acid-Vibratome technique, which utilizes fresh, glyoxylic acid-perfused tissue. Thus, the preterminal axon pathways become fluorescent throughout their full extent and the several new terminal systems, discovered with the glyoxylic acid-Vibratome method, are well demonstrable. The method is also highly useful for the study of the cell bodies and their dendritic processes. The catacholamine fibre systems are visualized without any signs of diffusion and with a richness in detail. In animals pretreated with L-tryptophan and MAO-inhibitor the technique is also useful for studies on central indolamine-containing systems. | [
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|
PMID:11201 | [Pharmacologic aspects in the management of inner ear disease (author's transl)]. | Our knowledge of the effects of drugs on the inner ear is extremely insufficient when compared to their effects on other organs. The role of drugs commonly recommended for therapy of inner ear disorders (such as, procaine, antihistaminics, diuretics, osmotics, corticoids and substances changing blood viscosity) has yet to be clarified since the clinical effects of these pharmaceuticals or vasoactive substances often cannot be confirmed by experimental study. Under present circumstances, the use of definitive therapy to prevent deafness has only limited application--as in thyroid hormone substitution to prevent deafness from cretinism. | [Pharmacologic aspects in the management of inner ear disease (author's transl)]. Our knowledge of the effects of drugs on the inner ear is extremely insufficient when compared to their effects on other organs. The role of drugs commonly recommended for therapy of inner ear disorders (such as, procaine, antihistaminics, diuretics, osmotics, corticoids and substances changing blood viscosity) has yet to be clarified since the clinical effects of these pharmaceuticals or vasoactive substances often cannot be confirmed by experimental study. Under present circumstances, the use of definitive therapy to prevent deafness has only limited application--as in thyroid hormone substitution to prevent deafness from cretinism. | [
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|
PMID:11205 | Thin layer chromatographic identification of some sympathomimetic amines. | Thin layer chromatographic behavior of some sympathomimetic amines in the presence of acids in neutral and organic solvent systems is reported. The sympathomimetic amines were dissolved in 0.1N HCl or ethanol and treated with bromocresol green or p-nitrobenzoyl chloride reagents on fiber sheets or precoated glass plates. Two-, 3-, and 4-, component solvent systems were tested. Benzene-ethyl acetate gave 2 spots for each amine standard; the more polar spots were satisfactorily separated. Amines in pharmaccuticals were not separated by any solvent system tested. | Thin layer chromatographic identification of some sympathomimetic amines. Thin layer chromatographic behavior of some sympathomimetic amines in the presence of acids in neutral and organic solvent systems is reported. The sympathomimetic amines were dissolved in 0.1N HCl or ethanol and treated with bromocresol green or p-nitrobenzoyl chloride reagents on fiber sheets or precoated glass plates. Two-, 3-, and 4-, component solvent systems were tested. Benzene-ethyl acetate gave 2 spots for each amine standard; the more polar spots were satisfactorily separated. Amines in pharmaccuticals were not separated by any solvent system tested. | [
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|
PMID:11206 | Evidence for a negative membrane potential and for movement of C1- against its electrochemical gradient in the ascomycete Neocosmospora vasinfecta. | The iodides of three lipid-soluble cations (dibenzyldimethylammonium; tribenzylmethylammonium, TBMA+; ethyldimethylbenzylammonium) were synthesized by the reaction of 14C-labeled methyl or 14C-labeled ethyl iodide with the appropriate secondary of tertiary amine and used in an attempt to measure the transmembrane electrical potential difference in Neocosmospora. Only mycelium containing high levels of Na+ accumulated measureable amounts of these cations and only above pH 6. Uptake was reduced in the presence of exogenous K+, Na+, Mg2+, or tris(hydroxymethyl)aminomethane. The velocity of TBMA+ uptake was proportional to its concentration between 46 and 427 muM. Neither the rate nor the extent of TBMB+ uptake was greatly affected by the presence of a fivefold excess of either dibenzyldimethylammonium or ethyldimethylbenzylammonium, even though these cations were themselves accumulated. The uncoupler m-chlorophenylhydrazone induced loss of previously accumulated TBMA+ from the mycelium. Anaerobiosis and cold (5 degrees C) temperature both inhibited TBMA+ uptake but did not induce the loss of previously accumulated TBMA+. The uptake of lipophilic cations by Na+-rich mycelium indicated a minimum transmembrane electrical potential of -60 to -70 mV (inside negative). Net uptake of these cations appeared to be strongly influenced by the availability of endogenous exchangeable cations and by the presence of other exogenous cations, as well as by the membrane potential. Despite these limitations, transport of C1- by Na+-rich mycelium appeared to take place against the electrochemical gradient for C1-. | Evidence for a negative membrane potential and for movement of C1- against its electrochemical gradient in the ascomycete Neocosmospora vasinfecta. The iodides of three lipid-soluble cations (dibenzyldimethylammonium; tribenzylmethylammonium, TBMA+; ethyldimethylbenzylammonium) were synthesized by the reaction of 14C-labeled methyl or 14C-labeled ethyl iodide with the appropriate secondary of tertiary amine and used in an attempt to measure the transmembrane electrical potential difference in Neocosmospora. Only mycelium containing high levels of Na+ accumulated measureable amounts of these cations and only above pH 6. Uptake was reduced in the presence of exogenous K+, Na+, Mg2+, or tris(hydroxymethyl)aminomethane. The velocity of TBMA+ uptake was proportional to its concentration between 46 and 427 muM. Neither the rate nor the extent of TBMB+ uptake was greatly affected by the presence of a fivefold excess of either dibenzyldimethylammonium or ethyldimethylbenzylammonium, even though these cations were themselves accumulated. The uncoupler m-chlorophenylhydrazone induced loss of previously accumulated TBMA+ from the mycelium. Anaerobiosis and cold (5 degrees C) temperature both inhibited TBMA+ uptake but did not induce the loss of previously accumulated TBMA+. The uptake of lipophilic cations by Na+-rich mycelium indicated a minimum transmembrane electrical potential of -60 to -70 mV (inside negative). Net uptake of these cations appeared to be strongly influenced by the availability of endogenous exchangeable cations and by the presence of other exogenous cations, as well as by the membrane potential. Despite these limitations, transport of C1- by Na+-rich mycelium appeared to take place against the electrochemical gradient for C1-. | [
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|
PMID:11207 | Unique aspects of the regulation of the aspartate transcarbamylase of Serratia marcescens. | Aspartate trancarbamylase (ATC ase; EC 2.1.3.2) from Serratia marcescens HY has been purified 134-fold. Its properties are unique. Unlike the ATCase from Escherichia coli and Salmonella typhimurium, the S. marcescens HY enzyme activity is not feedback inhibited by any purine or pyrimidine nucleotide effectors; instead, the enzyme is activated by both cytidine 5'-triphosphate and adenosine 5'-triphosphate. Like the ATCase from E. coli and S. typhimurium, adenosine 5'-triphosphate alters the [S]0.5 of the enzyme and, in contrast, cytidine 5'-triphosphate does not alter the [S]0.5 but, instead, alters the Vmax. As has been shown for both E. coli and S . typhimurium, effector sensitivity may be selectively dissociated form catalytic activity by treatment with heat, parachloromercuribenzoate, or neohydrin. This dissociated enzyme possesses threefold higher specific activity than the native enzyme. The sedimentation coefficient of the native enzyme is approximately 11.4S, whereas the dissociated enzyme has a value of 6.0S. Whereas it has been possible to reconstitute the E. coli and the S. marcescens ATCase enzymes from their own homologous subunits, it has not been possible to make hybrid enzymes of catalytic and regulatory heterologous subunits from each other. It was not possible to detect repression of ATCase formation after growth of prototrophic strains of S. marcescens HY supplemented with 200 mug of uracil per ml, but eightfold derepression was observed after uracil withdrawal in pyrimidine auxotrophs. | Unique aspects of the regulation of the aspartate transcarbamylase of Serratia marcescens. Aspartate trancarbamylase (ATC ase; EC 2.1.3.2) from Serratia marcescens HY has been purified 134-fold. Its properties are unique. Unlike the ATCase from Escherichia coli and Salmonella typhimurium, the S. marcescens HY enzyme activity is not feedback inhibited by any purine or pyrimidine nucleotide effectors; instead, the enzyme is activated by both cytidine 5'-triphosphate and adenosine 5'-triphosphate. Like the ATCase from E. coli and S. typhimurium, adenosine 5'-triphosphate alters the [S]0.5 of the enzyme and, in contrast, cytidine 5'-triphosphate does not alter the [S]0.5 but, instead, alters the Vmax. As has been shown for both E. coli and S . typhimurium, effector sensitivity may be selectively dissociated form catalytic activity by treatment with heat, parachloromercuribenzoate, or neohydrin. This dissociated enzyme possesses threefold higher specific activity than the native enzyme. The sedimentation coefficient of the native enzyme is approximately 11.4S, whereas the dissociated enzyme has a value of 6.0S. Whereas it has been possible to reconstitute the E. coli and the S. marcescens ATCase enzymes from their own homologous subunits, it has not been possible to make hybrid enzymes of catalytic and regulatory heterologous subunits from each other. It was not possible to detect repression of ATCase formation after growth of prototrophic strains of S. marcescens HY supplemented with 200 mug of uracil per ml, but eightfold derepression was observed after uracil withdrawal in pyrimidine auxotrophs. | [
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|
PMID:11208 | Reduced nicotinamide adenine dinucleotide oxidase activity in membranes and cytoplasm of Acholeplasma laidlawii and Mycoplasma mycoides subsp. capri. | The properties of the membrane-bound reduced nicotinamide adenine dinucleotide (NADH) oxidase of Acholeplasma laidlawii were compared with those of the corresponding cytoplasmic activity of Mycoplasma mycoides subsp. capri. The striking differences in pH optima, susceptibility to inhibitors and detergents, and heat inactivation between the NADH oxidase activity, with oxygen as an electron acceptor, and the NADH oxidoreductase activity, with dichlorophenol indophenol (DCPIP) as an alternate electron acceptor, support the presence of more than one catalytic protein in both the membrane-bound and soluble enzyme systems. The detection of more than one band positive for the NADH-nitroblue tetrazolium oxidoreductase reaction on electrophoresis of either the membranes of A. laidlawii or the cytoplasm of M mycoides subsp. capri also points in the same direction. The membrane-bound enzyme system differed, however, form the soluble one because it had a lower ratio of oxidase activity to oxidoreductase activity, and because it was less susceptible to heat inactivation and more readily incorporated incorporated into reaggregated membranes. In addition, the specific activity of the membrane-bound enzyme system increased as the culture aged, whereas that of the soluble system decreased as the culture aged. It is suggested that the different location in the cell could be responsible for some of the differences between the membrane-bound NADH oxidase activity of A. laidlawii and that found in the cytoplasm of M. mycoides subsp. capri. | Reduced nicotinamide adenine dinucleotide oxidase activity in membranes and cytoplasm of Acholeplasma laidlawii and Mycoplasma mycoides subsp. capri. The properties of the membrane-bound reduced nicotinamide adenine dinucleotide (NADH) oxidase of Acholeplasma laidlawii were compared with those of the corresponding cytoplasmic activity of Mycoplasma mycoides subsp. capri. The striking differences in pH optima, susceptibility to inhibitors and detergents, and heat inactivation between the NADH oxidase activity, with oxygen as an electron acceptor, and the NADH oxidoreductase activity, with dichlorophenol indophenol (DCPIP) as an alternate electron acceptor, support the presence of more than one catalytic protein in both the membrane-bound and soluble enzyme systems. The detection of more than one band positive for the NADH-nitroblue tetrazolium oxidoreductase reaction on electrophoresis of either the membranes of A. laidlawii or the cytoplasm of M mycoides subsp. capri also points in the same direction. The membrane-bound enzyme system differed, however, form the soluble one because it had a lower ratio of oxidase activity to oxidoreductase activity, and because it was less susceptible to heat inactivation and more readily incorporated incorporated into reaggregated membranes. In addition, the specific activity of the membrane-bound enzyme system increased as the culture aged, whereas that of the soluble system decreased as the culture aged. It is suggested that the different location in the cell could be responsible for some of the differences between the membrane-bound NADH oxidase activity of A. laidlawii and that found in the cytoplasm of M. mycoides subsp. capri. | [
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|
PMID:11210 | Calcium transport ATPase of canine cardiac sarcoplasmic reticulum. A comparison with that of rabbit fast skeletal muscle sarcoplasmic reticulum. | To define the mechanism responsible for the slow rate of calcium transport by cardiac sarcoplasmic reticulum, the kinetic properties of the Ca2+-dependent ATPase of canine cardiac microsomes were characterized and compared with those of a comparable preparation from rabbit fast skeletal muscle. A phosphoprotein intermediate (E approximately P), which has the stability characteristics of an acyl phosphate, is formed during ATP hydrolysis by cardiac microsomes. Ca2+ is required for the E approximately P formation, and Mg2+ accelerates its decomposition. The Ca2+ concentration required for half-maximal activation of the ATPase is 4.7 +/- 0.2 muM for cardiac microsomes and 1.3 +/- 0.1 muM for skeletal microsomes at pH 6.8 and 0 degrees. The ATPase activities at saturating concentrations of ionized Ca2+ and pH 6.8, expressed as ATP hydrolysis per mg of protein, are 3 to 6 times lower for cardiac microsomes than for skeletal microsomes under a variety of conditions tested. The apparent Km value for MgATP at high concentrations in the presence of saturating concentrations of ionized Ca2+ is 0.18 +/- 0.03 ms at pH 6.8 and 25 degrees. The maximum velocity of ATPase activity under these conditions is 0.45 +/- 0.05 mumol per mg per min for cardiac microsomes and 1.60 +/- 0.05 mumol per mg per min for skeletal microsomes. The maximum steady state level of E approximately P for cardiac microsomes, 1.3 +/- 0.1 nmol per mg, is significantly less than the value of 4.9 +/- 0.2 nmol per mg for skeletal microsomes, so that the turnover number of the Ca2+-dependent ATPase of cardiac microsomes, calculated as the ratio of ATPase activity to the E approximately P level is similar to that of the skeletal ATPase. These findings indicate that the relatively slow rate of calcium transport by cardiac microsomes, whem compared to that of skeletal microsomes, reflects a lower density of calcium pumping sites and lower Ca2+ affinity for these sites, rather than a lower turnover rate. | Calcium transport ATPase of canine cardiac sarcoplasmic reticulum. A comparison with that of rabbit fast skeletal muscle sarcoplasmic reticulum. To define the mechanism responsible for the slow rate of calcium transport by cardiac sarcoplasmic reticulum, the kinetic properties of the Ca2+-dependent ATPase of canine cardiac microsomes were characterized and compared with those of a comparable preparation from rabbit fast skeletal muscle. A phosphoprotein intermediate (E approximately P), which has the stability characteristics of an acyl phosphate, is formed during ATP hydrolysis by cardiac microsomes. Ca2+ is required for the E approximately P formation, and Mg2+ accelerates its decomposition. The Ca2+ concentration required for half-maximal activation of the ATPase is 4.7 +/- 0.2 muM for cardiac microsomes and 1.3 +/- 0.1 muM for skeletal microsomes at pH 6.8 and 0 degrees. The ATPase activities at saturating concentrations of ionized Ca2+ and pH 6.8, expressed as ATP hydrolysis per mg of protein, are 3 to 6 times lower for cardiac microsomes than for skeletal microsomes under a variety of conditions tested. The apparent Km value for MgATP at high concentrations in the presence of saturating concentrations of ionized Ca2+ is 0.18 +/- 0.03 ms at pH 6.8 and 25 degrees. The maximum velocity of ATPase activity under these conditions is 0.45 +/- 0.05 mumol per mg per min for cardiac microsomes and 1.60 +/- 0.05 mumol per mg per min for skeletal microsomes. The maximum steady state level of E approximately P for cardiac microsomes, 1.3 +/- 0.1 nmol per mg, is significantly less than the value of 4.9 +/- 0.2 nmol per mg for skeletal microsomes, so that the turnover number of the Ca2+-dependent ATPase of cardiac microsomes, calculated as the ratio of ATPase activity to the E approximately P level is similar to that of the skeletal ATPase. These findings indicate that the relatively slow rate of calcium transport by cardiac microsomes, whem compared to that of skeletal microsomes, reflects a lower density of calcium pumping sites and lower Ca2+ affinity for these sites, rather than a lower turnover rate. | [
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PMID:11211 | Purification and characterization of L-asparaginase with anti-lymphoma activity from Vibrio succinogenes. | Homogeneols L-asparaginase with anti-lymphoma activity was prepared from Vibrio succinogenes, an anaerobic bacterium from the bovine rumen. An overall yield of pure L-asparaginase of 40 to 45% and a specific activity of 200 +/- 2 IU per mg of protein was obtained. The pure enzyme can be stored at -20 degrees for at least 3 months with no loss of activity. The isoelectric point of the L-asparaginase is 8.74. No carbohydrate, phosphorus, tryptophan, disulfide, or sulfhydryl groups were detected. The enzyme has a molecular weight of 146,000 and a subunit weight of approximately 37,000. The Km of the enzyme for L-asparagine is 4.78 X 10(-5) M and the pH optimum of the L-asparaginase reaction is 7.3. D-Asparagine was hydrolyzed at 6.5% of the rate found with the L isomer. L-Glutamine and a variety of other amides were not hydrolyzed at significant rates; the activity of the enzyme for L-glutamine was 130- to 600-fold less than that of other therapeutically effective L-asparaginases of bacterial origin. The L-asparaginase from V. succinogenes is immunologically distinct from the L-asparaginase (EC-2) of Escherichia coli. | Purification and characterization of L-asparaginase with anti-lymphoma activity from Vibrio succinogenes. Homogeneols L-asparaginase with anti-lymphoma activity was prepared from Vibrio succinogenes, an anaerobic bacterium from the bovine rumen. An overall yield of pure L-asparaginase of 40 to 45% and a specific activity of 200 +/- 2 IU per mg of protein was obtained. The pure enzyme can be stored at -20 degrees for at least 3 months with no loss of activity. The isoelectric point of the L-asparaginase is 8.74. No carbohydrate, phosphorus, tryptophan, disulfide, or sulfhydryl groups were detected. The enzyme has a molecular weight of 146,000 and a subunit weight of approximately 37,000. The Km of the enzyme for L-asparagine is 4.78 X 10(-5) M and the pH optimum of the L-asparaginase reaction is 7.3. D-Asparagine was hydrolyzed at 6.5% of the rate found with the L isomer. L-Glutamine and a variety of other amides were not hydrolyzed at significant rates; the activity of the enzyme for L-glutamine was 130- to 600-fold less than that of other therapeutically effective L-asparaginases of bacterial origin. The L-asparaginase from V. succinogenes is immunologically distinct from the L-asparaginase (EC-2) of Escherichia coli. | [
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PMID:11212 | Hydrogen ion interactions of horse spleen ferritin and apoferritin. | The interactions of horse spleen ferritin and its derivative apoferritin with H+ ions were studied by potentiometric and spectrophotometric titration; to aid in data analysis, heats of ionization over a limited pH range and amide content were also determined. Per apoferritin subunit, all tyrosine and cysteine side chains, two of the nine lysine side chains and at least three of the six histidine side chains were found not to titrate; a preliminary but self-consistent analysis of the titration data is proposed. The titration curve of ferritin was identical with that of apoferritin in the pH range 5.5 to 3. In addition, under the conditions used, the reactivities of ferritin histidines to bromoacetate and of ferritin lysines to formaldehyde were identical with those in apoferritin. Above pH 8, a time-dependent titration of the ferritin core occurs which prevents comparison of the titration curves of the two proteins in this region. However, in the pH regions 5.5 to 7.5, two extra groups per subunit titrate reversibly in ferritin relative to apoferritin. Moreover, although the isoionic points of ferritin and apoferritin are identical in water, the isoionic point of ferritin is 0.5 pH unit lower than that of apoferritin in 0.16 to 1 M KCl. The different effects of KCl and NaCl on the two proteins indicate the presence of cation binding sites in ferritin that are absent in apoferritin and possibly also the presence of anion binding sites in apoferritin that are occupied in ferritin by anions of the core. The difference between the isoionic points of the two proteins in KCl has been interpreted to indicate the presence of approximately 2 phosphate residues per ferritin subunit which serve as cation binding sites and which are negatively charged at the isoionic point in KCl. These phosphates may also represent the additional residues that titrate in ferritin between pH 5.5 and 7.5, or may interact with positively charged residues on the inner surface of the ferritin shell, or both. | Hydrogen ion interactions of horse spleen ferritin and apoferritin. The interactions of horse spleen ferritin and its derivative apoferritin with H+ ions were studied by potentiometric and spectrophotometric titration; to aid in data analysis, heats of ionization over a limited pH range and amide content were also determined. Per apoferritin subunit, all tyrosine and cysteine side chains, two of the nine lysine side chains and at least three of the six histidine side chains were found not to titrate; a preliminary but self-consistent analysis of the titration data is proposed. The titration curve of ferritin was identical with that of apoferritin in the pH range 5.5 to 3. In addition, under the conditions used, the reactivities of ferritin histidines to bromoacetate and of ferritin lysines to formaldehyde were identical with those in apoferritin. Above pH 8, a time-dependent titration of the ferritin core occurs which prevents comparison of the titration curves of the two proteins in this region. However, in the pH regions 5.5 to 7.5, two extra groups per subunit titrate reversibly in ferritin relative to apoferritin. Moreover, although the isoionic points of ferritin and apoferritin are identical in water, the isoionic point of ferritin is 0.5 pH unit lower than that of apoferritin in 0.16 to 1 M KCl. The different effects of KCl and NaCl on the two proteins indicate the presence of cation binding sites in ferritin that are absent in apoferritin and possibly also the presence of anion binding sites in apoferritin that are occupied in ferritin by anions of the core. The difference between the isoionic points of the two proteins in KCl has been interpreted to indicate the presence of approximately 2 phosphate residues per ferritin subunit which serve as cation binding sites and which are negatively charged at the isoionic point in KCl. These phosphates may also represent the additional residues that titrate in ferritin between pH 5.5 and 7.5, or may interact with positively charged residues on the inner surface of the ferritin shell, or both. | [
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PMID:11213 | Purification and properties of two aromatic aminotransferases in Bacillus subtilis. | Two enzymes which transaminate tyrosine and phenylalanine in Bacillus subtilis were each purified over 200-fold and partially characterized. One of the enzymes, termed histidinol phosphate aminotransferase, is also active with imidazole acetyl phosphate as the amino group recipient. Previous studies have shown that mutants lacking this enzyme require histidine for growth. Mutants in the other enzyme termed aromatic aminotransferase are prototrophs. Neither enzyme is active on any other substrate involved in amino acid synthesis. The two enzymes can be distinguished by a number of criteria. Gel filtration analysis indicate the aromatic and histidinol phosphate aminotransferases have molecular weights of 63,500 and 33,000, respectively. Histidinol phosphate aminotransferase is heat-sensitive, whereas aromatic aminotransferase is relatively heat-stable, particularly in the presence of alpha-ketoglutarate. Both enzymes display typical Michaelis-Menten kinetics in their rates of reaction. The two enzymes have similar pH optima and employ a ping-pong mechanism of action. The Km values for various substrates suggest that histidinol phosphate aminotransferase is the predominant enzyme responsible for the transamaination reactions in the synthesis of tyrosine and phenylalanine. This enzyme has a 4-fold higher affinity for tyrosine and phenylalanine than does the aromatic aminotransferase. Competitive substrate inhibition was observed between tyrosine, phenylalanine, and histidinol phosphate for histidinol phosphate aminotransferase. The significance of the fact that an enzyme of histidine synthesis plays an important role in aromatic amino acid synthesis is discussed. | Purification and properties of two aromatic aminotransferases in Bacillus subtilis. Two enzymes which transaminate tyrosine and phenylalanine in Bacillus subtilis were each purified over 200-fold and partially characterized. One of the enzymes, termed histidinol phosphate aminotransferase, is also active with imidazole acetyl phosphate as the amino group recipient. Previous studies have shown that mutants lacking this enzyme require histidine for growth. Mutants in the other enzyme termed aromatic aminotransferase are prototrophs. Neither enzyme is active on any other substrate involved in amino acid synthesis. The two enzymes can be distinguished by a number of criteria. Gel filtration analysis indicate the aromatic and histidinol phosphate aminotransferases have molecular weights of 63,500 and 33,000, respectively. Histidinol phosphate aminotransferase is heat-sensitive, whereas aromatic aminotransferase is relatively heat-stable, particularly in the presence of alpha-ketoglutarate. Both enzymes display typical Michaelis-Menten kinetics in their rates of reaction. The two enzymes have similar pH optima and employ a ping-pong mechanism of action. The Km values for various substrates suggest that histidinol phosphate aminotransferase is the predominant enzyme responsible for the transamaination reactions in the synthesis of tyrosine and phenylalanine. This enzyme has a 4-fold higher affinity for tyrosine and phenylalanine than does the aromatic aminotransferase. Competitive substrate inhibition was observed between tyrosine, phenylalanine, and histidinol phosphate for histidinol phosphate aminotransferase. The significance of the fact that an enzyme of histidine synthesis plays an important role in aromatic amino acid synthesis is discussed. | [
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|
PMID:11214 | A new oxygenase, 2-nitropropane dioxygenase of Hansenula mrakii. Enzymologic and spectrophotometric properties. | 2-Nitropropane dioxygenase, purified to homogeneity from Hansenula mrakii (IFO 0895), has a molecular weight of approximately 62,000 and consists of two subunits nonidentical in molecular weight (39,000 and 25,000). Stoichiometrical studies and the results obtained with 18O2 showed that 2 atoms of molecular oxygen are incorporated into 2 molecules of acetone formed from 2-nitropropane. In addition to 2-nitropropane, nitroethane, 3-nitro-2-pentanol, and 1-nitropropane are oxidatively dentrified. The enzyme, which exhibits absorption maxima at 274, 370, 415, and 440 nm and a shoulder at 470 nm, contains 1 mol of FAD and 1 g atom of non-heme iron per mol of enzyme. The enzyme-bound FAD is reduced by 2-nitropropane under anaerogic conditions, but the enzyme-bound Fe3+ is not affected. The introduction of oxygen to the reduced form of enzyme causes reoxidation of the enzyme. The bound FAD and Fe3+ are reduced by the addition of nitromethane, which is not a substrate, under anaerobic conditions. The aerobic dialysis of the enzyme treated with nitromethane causes reoxidation of only the Fe2+. Sodium dithionite also reduces both the enzyme-bound FAD and Fe3+ under anaerobic conditions. When the enzyme is dialyzed against 10 mM potassium phosphate buffer (pH 7.0) immediately after reduction by dithionite, the absorption spectrum similar to that of the native enzyme appeared with concomitant restoration of approximately 80% of the activity. The enzyme activity is significantly inhibited by pyrocatechol-3,5-disulfonate disodium salt, 8-hydroxyquinoline, reducing agents such as 2-mercaptoethanol, and HgCl2. The Michaelis constants are as follows: 2-nitropropane (2.13 X 10(-2) M), nitroethane (2.43 X 10(-2) M), 3-nitro-2-pentanol (6.8 X 10(-3) M), 1-nitropropane (2.56 X 10(-2) M), and oxygen (3.03 X 10(-4) M, with 2-nitropropane). | A new oxygenase, 2-nitropropane dioxygenase of Hansenula mrakii. Enzymologic and spectrophotometric properties. 2-Nitropropane dioxygenase, purified to homogeneity from Hansenula mrakii (IFO 0895), has a molecular weight of approximately 62,000 and consists of two subunits nonidentical in molecular weight (39,000 and 25,000). Stoichiometrical studies and the results obtained with 18O2 showed that 2 atoms of molecular oxygen are incorporated into 2 molecules of acetone formed from 2-nitropropane. In addition to 2-nitropropane, nitroethane, 3-nitro-2-pentanol, and 1-nitropropane are oxidatively dentrified. The enzyme, which exhibits absorption maxima at 274, 370, 415, and 440 nm and a shoulder at 470 nm, contains 1 mol of FAD and 1 g atom of non-heme iron per mol of enzyme. The enzyme-bound FAD is reduced by 2-nitropropane under anaerogic conditions, but the enzyme-bound Fe3+ is not affected. The introduction of oxygen to the reduced form of enzyme causes reoxidation of the enzyme. The bound FAD and Fe3+ are reduced by the addition of nitromethane, which is not a substrate, under anaerobic conditions. The aerobic dialysis of the enzyme treated with nitromethane causes reoxidation of only the Fe2+. Sodium dithionite also reduces both the enzyme-bound FAD and Fe3+ under anaerobic conditions. When the enzyme is dialyzed against 10 mM potassium phosphate buffer (pH 7.0) immediately after reduction by dithionite, the absorption spectrum similar to that of the native enzyme appeared with concomitant restoration of approximately 80% of the activity. The enzyme activity is significantly inhibited by pyrocatechol-3,5-disulfonate disodium salt, 8-hydroxyquinoline, reducing agents such as 2-mercaptoethanol, and HgCl2. The Michaelis constants are as follows: 2-nitropropane (2.13 X 10(-2) M), nitroethane (2.43 X 10(-2) M), 3-nitro-2-pentanol (6.8 X 10(-3) M), 1-nitropropane (2.56 X 10(-2) M), and oxygen (3.03 X 10(-4) M, with 2-nitropropane). | [
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|
PMID:11215 | Reconstitution of biological molecular generators of electric current. H+-ATPase. | 1. Generation of a transmembrane electric potential difference by oligomycin-sensitive ATPase complex, incorporated into spherical or planar phospholipid membrane, has been demonstrated. To this end, penetrating anion probe and direct voltmeter measurement of electric potential across phospholipid membrane were used. It was found that ATP-induced electric response is sensitive to oligomycin and protonophorous uncouplers. 2. The effect of variations in the phospholipid component of proteoliposomes on the electric generation was studied. It was revealed that the usage of mitochondrial phospholipids and phosphatidylethanolamine allows the highest values of membrane potential to be obtained in the case of ATPase proteoliposomes. In the case of cytochrome oxidase and bacteriorhodopsin proteoliposomes, phosphatidylserine was also shown to be quite suitable. Phosphatidylcholine was absolutely ineffective in all cases. 3. In proteoliposomes, containing both ATPase and bacteriorhodopsin, ATP and light induced generation of the electric field of the same direction. 4. In ATPase + cytochrome oxidase proteoliposomes, ATP hydrolysis and ascorbate oxidation was found to support electric generation of the same direction if cytochrome c was inside vesicles. Oxidation via external cytochrome c resulted in formation of electric field of the direction, opposite to that induced by ATP hydrolysis. 5. The data obtained in experiments with proteoliposomes of different types are discussed. The conclusion is made that conversion of energy of different resources into electric form is a common feature of membraneous energy transducers, which is in agreement with the Mitchellian principle of cellular energetics. | Reconstitution of biological molecular generators of electric current. H+-ATPase. 1. Generation of a transmembrane electric potential difference by oligomycin-sensitive ATPase complex, incorporated into spherical or planar phospholipid membrane, has been demonstrated. To this end, penetrating anion probe and direct voltmeter measurement of electric potential across phospholipid membrane were used. It was found that ATP-induced electric response is sensitive to oligomycin and protonophorous uncouplers. 2. The effect of variations in the phospholipid component of proteoliposomes on the electric generation was studied. It was revealed that the usage of mitochondrial phospholipids and phosphatidylethanolamine allows the highest values of membrane potential to be obtained in the case of ATPase proteoliposomes. In the case of cytochrome oxidase and bacteriorhodopsin proteoliposomes, phosphatidylserine was also shown to be quite suitable. Phosphatidylcholine was absolutely ineffective in all cases. 3. In proteoliposomes, containing both ATPase and bacteriorhodopsin, ATP and light induced generation of the electric field of the same direction. 4. In ATPase + cytochrome oxidase proteoliposomes, ATP hydrolysis and ascorbate oxidation was found to support electric generation of the same direction if cytochrome c was inside vesicles. Oxidation via external cytochrome c resulted in formation of electric field of the direction, opposite to that induced by ATP hydrolysis. 5. The data obtained in experiments with proteoliposomes of different types are discussed. The conclusion is made that conversion of energy of different resources into electric form is a common feature of membraneous energy transducers, which is in agreement with the Mitchellian principle of cellular energetics. | [
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|
PMID:11216 | Mechanism of intramolecular activation of pepsinogen. Evidence for an intermediate delta and the involvement of the active site of pepsin in the intramolecular activation of pepsinogen. | Intramolecular pepsinogen activation is inhibited either by pepstatin, a potent pepsin inhibitor, or by purified globin from hemoglobin, a good pepsin substrate. Also, pepsinogen at pH 2 can be bound to a pepstatin-Sepharose column and recovered as native zymogen upon elution in pH 8 buffer. Kinetic studies of the globin inhibition of pepsinogen activation show that globin binds to a pepsinogen intermediate. This interaction gives rise to competitive inhibition of intramolecular pepsinogen activation. The evidence presented in this paper suggests that pepsinogen is converted rapidly upon acidification to the pepsinogen intermediate delta. In the absence of an inhibitor, the intermediate undergoes conformational change to bind the activation peptide portion of this same pepsinogen molecule in the active center to form an intramolecular enzyme-substrate complex (intermediate theta). This is followed by the intramolecular hydrolysis of the peptide bond between residues 44 and 45 of the pepsinogen molecule and the dissociation of the activation peptide from the pepsin. Intermediate delta apparently does not activate another pepsinogen molecule via an intermolecular process. Neither does intermediate delta hydrolyze globin substrate. | Mechanism of intramolecular activation of pepsinogen. Evidence for an intermediate delta and the involvement of the active site of pepsin in the intramolecular activation of pepsinogen. Intramolecular pepsinogen activation is inhibited either by pepstatin, a potent pepsin inhibitor, or by purified globin from hemoglobin, a good pepsin substrate. Also, pepsinogen at pH 2 can be bound to a pepstatin-Sepharose column and recovered as native zymogen upon elution in pH 8 buffer. Kinetic studies of the globin inhibition of pepsinogen activation show that globin binds to a pepsinogen intermediate. This interaction gives rise to competitive inhibition of intramolecular pepsinogen activation. The evidence presented in this paper suggests that pepsinogen is converted rapidly upon acidification to the pepsinogen intermediate delta. In the absence of an inhibitor, the intermediate undergoes conformational change to bind the activation peptide portion of this same pepsinogen molecule in the active center to form an intramolecular enzyme-substrate complex (intermediate theta). This is followed by the intramolecular hydrolysis of the peptide bond between residues 44 and 45 of the pepsinogen molecule and the dissociation of the activation peptide from the pepsin. Intermediate delta apparently does not activate another pepsinogen molecule via an intermolecular process. Neither does intermediate delta hydrolyze globin substrate. | [
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|
PMID:11217 | Temperature dependence of cholesterol binding to cytochrome P-450scc of the rat adrenal. Effect of adrenocorticotropic hormone and cycloheximide. | A type I absorbance change is observed in suspensions of adrenal cortical mitochondria as the temperature is increased from 0-22 degrees. This "heat-generated" type I absorbance change is similar in magnitude to the pregnenolone-induced type II absorbance change of these mitochondria. Studies with inhibitors of cholesterol side chain cleavage indicate that the heat-generated type I absorbance change represents the specific interaction of cytochrome P-450scc with endogenous cholesterol in the mitochondria. This finding is confirmed by low temperature EPR spectroscopy on temperature-equilibrated, quick frozen adrenal mitochondrial samples. The EPR resonance at g = 8.2, which is that of the high spin cholesterol-bound cytochrome P-450scc, is absent in the samples incubated at 0 degrees and increases in magnitude with increasing temperature of incubation. Studies of the pH dependence of the heat-generated type I and pregnenolone-induced type II absorbance changes reveal that both are diminished by increasing pH over the range 6 to 8. Adrenocorticotropic hormone (ACTH) treatment of rats results in adrenal mitochondria which show a greatly increased heat-generated type I absorbance change. The latter correlates with an increased pregnenolone-induced type II absorbance change and increased EPR g = 8.2 signal. Prior treatment of animals with cycloheximide eliminated the ACTH-induced increase in the heat-generated type I absorbance change, the pregnenolone-induced type II absorbance change and the EPR g = 8.2 signal. We estimate that the hydrophobic bonding of cholesterol to cytochrome P-450scc occurs with a deltaH0' of approximately +15 kcal/mol and a deltaS0' of approximately +55 cal/mol deg. Our data support the concept of a labile protein which participates directly in this process. | Temperature dependence of cholesterol binding to cytochrome P-450scc of the rat adrenal. Effect of adrenocorticotropic hormone and cycloheximide. A type I absorbance change is observed in suspensions of adrenal cortical mitochondria as the temperature is increased from 0-22 degrees. This "heat-generated" type I absorbance change is similar in magnitude to the pregnenolone-induced type II absorbance change of these mitochondria. Studies with inhibitors of cholesterol side chain cleavage indicate that the heat-generated type I absorbance change represents the specific interaction of cytochrome P-450scc with endogenous cholesterol in the mitochondria. This finding is confirmed by low temperature EPR spectroscopy on temperature-equilibrated, quick frozen adrenal mitochondrial samples. The EPR resonance at g = 8.2, which is that of the high spin cholesterol-bound cytochrome P-450scc, is absent in the samples incubated at 0 degrees and increases in magnitude with increasing temperature of incubation. Studies of the pH dependence of the heat-generated type I and pregnenolone-induced type II absorbance changes reveal that both are diminished by increasing pH over the range 6 to 8. Adrenocorticotropic hormone (ACTH) treatment of rats results in adrenal mitochondria which show a greatly increased heat-generated type I absorbance change. The latter correlates with an increased pregnenolone-induced type II absorbance change and increased EPR g = 8.2 signal. Prior treatment of animals with cycloheximide eliminated the ACTH-induced increase in the heat-generated type I absorbance change, the pregnenolone-induced type II absorbance change and the EPR g = 8.2 signal. We estimate that the hydrophobic bonding of cholesterol to cytochrome P-450scc occurs with a deltaH0' of approximately +15 kcal/mol and a deltaS0' of approximately +55 cal/mol deg. Our data support the concept of a labile protein which participates directly in this process. | [
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|
PMID:11218 | 6-Phosphogluconate dehydrogenase. Purification and kinetics. | A method is described for the isolation and purification of 6-phosphogluconate dehydrogenase from pig liver. The molecular weight is estimated at 83,000 and that of the subunits is 42,000 as determined by gel electrophoresis. The pH maximum is 8.5 in 50 mM glycine/NaOH buffer and from 7.5 to 10 in 50 mM phosphate buffer at 30 degrees. Magnesium ion is not required for activity and acts as an inhibitor at concentrations above 20 mM. A cellular fractionation study indicates that this enzyme is located almost entirely within the soluble portion of the cytoplasm. Kinetic studies have been done in 50 mM glycine buffer, pH 8.5, at 30 degrees. The data are consistent with a sequential mechanism in which NADP+ is added first, followed by 6-phosphogluconate, and the products are released in the order, CO2, ribulose 5-phosphate, and NADPH. The Michaelis constant is 13.5 muM for 6-phosphogluconate. Dissociation constants are 4.8 muM for NADP+ and 5.1 muM for NADPH. | 6-Phosphogluconate dehydrogenase. Purification and kinetics. A method is described for the isolation and purification of 6-phosphogluconate dehydrogenase from pig liver. The molecular weight is estimated at 83,000 and that of the subunits is 42,000 as determined by gel electrophoresis. The pH maximum is 8.5 in 50 mM glycine/NaOH buffer and from 7.5 to 10 in 50 mM phosphate buffer at 30 degrees. Magnesium ion is not required for activity and acts as an inhibitor at concentrations above 20 mM. A cellular fractionation study indicates that this enzyme is located almost entirely within the soluble portion of the cytoplasm. Kinetic studies have been done in 50 mM glycine buffer, pH 8.5, at 30 degrees. The data are consistent with a sequential mechanism in which NADP+ is added first, followed by 6-phosphogluconate, and the products are released in the order, CO2, ribulose 5-phosphate, and NADPH. The Michaelis constant is 13.5 muM for 6-phosphogluconate. Dissociation constants are 4.8 muM for NADP+ and 5.1 muM for NADPH. | [
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|
PMID:11219 | Energy-dependent calcium transport in endoplasmic reticulum of adipocytes. | The endoplasmic reticulum from isolated rat adipocytes has the ability to actively accumulate calcium. The calcium uptake was characterized using the 20,000 X g supernatant (S1 fraction) of total cellular homogenate. Endoplasmic reticulum vesicles isolated from the S1 fraction as a 160,000 X g microsomal pellet prior to testing demonstrated little ability to accumulate calcium. The calcium uptake in the S1 fraction was localized to the endoplasmic reticulum vesicles by morphologic appearance, by the use of selective inhibitors of calcium uptake, and by high speed sedimentation of the accumulated calcium. The uptake was MgATP- and temperature-dependent and was sustained by the oxalate used as the intravesicular trapping agent. Uptake was linear with time for at least 30 min at all calcium concentrations tested (3 to 100 muM) and exhibited a pH optimum of approximately 7.0. The sulfhydryl inhibitor p-chloromercuribenzene sulfonate produced a dose-dependent inhibition of calcium uptake with total inhibition at 0.07 mumol/mg protein. Ruthenium red and sodium azide inhibited less than 5% of the uptake at concentrations (5 muM and 10 mM, respectively) which completely blocked calcium uptake by mitochondria isolated from the same cells. The Km for calcium uptake was 10 muM total calcium which corresponded to approximately 3.6 muM ionized calcium in the assay system. The maximum velocity of the uptake was 5.0 nmol (mg of microsomal protein)-1 (min)-1 at 24 degrees under the assay conditions used and exhibited a Q10 of 1.8. The uptake activity of the endoplasmic reticulum vesicles in the S1 fraction exhibited a marked time- and temperature-dependent lability which might account in part for the lack of uptake in the isolated microsomal fraction. This energy-dependent calcium uptake system would appear to be of physiologic importance to the regulation of intracellular calcium. | Energy-dependent calcium transport in endoplasmic reticulum of adipocytes. The endoplasmic reticulum from isolated rat adipocytes has the ability to actively accumulate calcium. The calcium uptake was characterized using the 20,000 X g supernatant (S1 fraction) of total cellular homogenate. Endoplasmic reticulum vesicles isolated from the S1 fraction as a 160,000 X g microsomal pellet prior to testing demonstrated little ability to accumulate calcium. The calcium uptake in the S1 fraction was localized to the endoplasmic reticulum vesicles by morphologic appearance, by the use of selective inhibitors of calcium uptake, and by high speed sedimentation of the accumulated calcium. The uptake was MgATP- and temperature-dependent and was sustained by the oxalate used as the intravesicular trapping agent. Uptake was linear with time for at least 30 min at all calcium concentrations tested (3 to 100 muM) and exhibited a pH optimum of approximately 7.0. The sulfhydryl inhibitor p-chloromercuribenzene sulfonate produced a dose-dependent inhibition of calcium uptake with total inhibition at 0.07 mumol/mg protein. Ruthenium red and sodium azide inhibited less than 5% of the uptake at concentrations (5 muM and 10 mM, respectively) which completely blocked calcium uptake by mitochondria isolated from the same cells. The Km for calcium uptake was 10 muM total calcium which corresponded to approximately 3.6 muM ionized calcium in the assay system. The maximum velocity of the uptake was 5.0 nmol (mg of microsomal protein)-1 (min)-1 at 24 degrees under the assay conditions used and exhibited a Q10 of 1.8. The uptake activity of the endoplasmic reticulum vesicles in the S1 fraction exhibited a marked time- and temperature-dependent lability which might account in part for the lack of uptake in the isolated microsomal fraction. This energy-dependent calcium uptake system would appear to be of physiologic importance to the regulation of intracellular calcium. | [
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|
PMID:11221 | A quantitative description of the extension and retraction of surface protrusions in spreading 3T3 mouse fibroblasts. | We suggest a method of quantitating the motile actions of surface protrusions in spreading animal cells in culture. Its basis is the determination of the percentage of freshly plated cells which produce particle-free areas around them on a gold particle-coated glass cover slip within 50 min. Studying 3T3 cells with this assay, we found that the presence of Na+, K+, Cl-, and Mg++ or Ca++ in a neutral or slightly alkaline phosphate or bicarbonate buffered solution is sufficient to support the optimal particle removal by the cells for at least 50 min. Two metabolic inhibitors, 2,4-dinitrophenol and Na-azide, inhibit the particle removal. If D-glucose is added along with the inhibitors, particle removal can be restored, whereas the addition of three glucose analogues which are generally believed to be nonmetabolizable cannot restore the activity. Serum is not required for the mechanism(s) of the motile actions of surface protrusions in spreading 3T3 cells. However, it contains components which can neutralize the inhibitory actions of bovine serum albumin and several amino acids, particularly L-cystine or L-cystein and L-methionine. Furthermore, serum codetermines which of the major surface extension, filopodia, lamellipodia, or lobopodia, is predominantly active. We found three distinct classes of extracellular conditions under which the active surface projections are predominantly either lamellipodia, (sheetlike projections), lobopodia (blebs), or filopodia (microspikes). The quantitated dependencies on temperature, pH and the inhibition by cytochalasin B or the particle removal are very similar in all three cases. Preventing the cells from anchoring themselves for 15-20 min before plating in serum-free medium seems to stimulate particle removal threefold. | A quantitative description of the extension and retraction of surface protrusions in spreading 3T3 mouse fibroblasts. We suggest a method of quantitating the motile actions of surface protrusions in spreading animal cells in culture. Its basis is the determination of the percentage of freshly plated cells which produce particle-free areas around them on a gold particle-coated glass cover slip within 50 min. Studying 3T3 cells with this assay, we found that the presence of Na+, K+, Cl-, and Mg++ or Ca++ in a neutral or slightly alkaline phosphate or bicarbonate buffered solution is sufficient to support the optimal particle removal by the cells for at least 50 min. Two metabolic inhibitors, 2,4-dinitrophenol and Na-azide, inhibit the particle removal. If D-glucose is added along with the inhibitors, particle removal can be restored, whereas the addition of three glucose analogues which are generally believed to be nonmetabolizable cannot restore the activity. Serum is not required for the mechanism(s) of the motile actions of surface protrusions in spreading 3T3 cells. However, it contains components which can neutralize the inhibitory actions of bovine serum albumin and several amino acids, particularly L-cystine or L-cystein and L-methionine. Furthermore, serum codetermines which of the major surface extension, filopodia, lamellipodia, or lobopodia, is predominantly active. We found three distinct classes of extracellular conditions under which the active surface projections are predominantly either lamellipodia, (sheetlike projections), lobopodia (blebs), or filopodia (microspikes). The quantitated dependencies on temperature, pH and the inhibition by cytochalasin B or the particle removal are very similar in all three cases. Preventing the cells from anchoring themselves for 15-20 min before plating in serum-free medium seems to stimulate particle removal threefold. | [
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|
PMID:11220 | Long-term results of mammary artery implants. | Eighty out of eighty-six patients (93%) with mammary artery implants were followed postoperatively for an average of three and a half years. The immediate mortality rate was 7% (6 cases), and the late mortality was 6% (5 cases). All had angina preoperatively. Twenty-four had a history of myocardial infarction and thirty-one were on limited physical activity, because of the pain. After surgery, thirty-three (45%) became asymptomatic. The angina improved significantly in thirty-five (47%) and remained unchanged in six (8%). Improvement in ventricular repolarization on ECG was observed in 69% of the patients. Postoperative cineangiography was performed in twenty-three patients; thirteen with single and ten with double implants. Out of the total of thirty-three implants, four (12%) were obstructed and twenty-seven patent (82%); twenty were in two cases of double implant, only one implant could be satisfactorily studied effectively functioning (61%). No obstructions were seen in the single implants. Non functioning implants were found in five (38%) of the thirteen single implants and in two of the twenty double ones (10%). The highest incidence of obstruction or non-functioning implants occurred in the group that did not show improvement (43%). This rate fell to 40% in the group that had some improvement and to 29% in those that were completely asymptomatic. Twelve of the eighteen patent mammary implants (67%) on the anterior wall of the left ventricle and eight of nine (89%) on the lateroinferior wall, established collateral circulation to the coronaries. Indication for surgery was considered satisfactory for nineteen out of the twenty-three patients and poor in four. There were two cases of obstruction of the implant (7%) in the group where surgery was correctly indicated and three of the twenty-three (11%) patent implants were non-functioning. Clinical improvement of the angina occurred in 84% in the first group and 50% in the other. In conclusion, this technique of indirect revascularization of the myocardium is valid for patients with severe diffuse lesions of the coronaries with a collateral network and preserved myocardial contractility. | Long-term results of mammary artery implants. Eighty out of eighty-six patients (93%) with mammary artery implants were followed postoperatively for an average of three and a half years. The immediate mortality rate was 7% (6 cases), and the late mortality was 6% (5 cases). All had angina preoperatively. Twenty-four had a history of myocardial infarction and thirty-one were on limited physical activity, because of the pain. After surgery, thirty-three (45%) became asymptomatic. The angina improved significantly in thirty-five (47%) and remained unchanged in six (8%). Improvement in ventricular repolarization on ECG was observed in 69% of the patients. Postoperative cineangiography was performed in twenty-three patients; thirteen with single and ten with double implants. Out of the total of thirty-three implants, four (12%) were obstructed and twenty-seven patent (82%); twenty were in two cases of double implant, only one implant could be satisfactorily studied effectively functioning (61%). No obstructions were seen in the single implants. Non functioning implants were found in five (38%) of the thirteen single implants and in two of the twenty double ones (10%). The highest incidence of obstruction or non-functioning implants occurred in the group that did not show improvement (43%). This rate fell to 40% in the group that had some improvement and to 29% in those that were completely asymptomatic. Twelve of the eighteen patent mammary implants (67%) on the anterior wall of the left ventricle and eight of nine (89%) on the lateroinferior wall, established collateral circulation to the coronaries. Indication for surgery was considered satisfactory for nineteen out of the twenty-three patients and poor in four. There were two cases of obstruction of the implant (7%) in the group where surgery was correctly indicated and three of the twenty-three (11%) patent implants were non-functioning. Clinical improvement of the angina occurred in 84% in the first group and 50% in the other. In conclusion, this technique of indirect revascularization of the myocardium is valid for patients with severe diffuse lesions of the coronaries with a collateral network and preserved myocardial contractility. | [
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|
PMID:11222 | Brush border motility. Microvillar contraction in triton-treated brush borders isolated from intestinal epithelium. | The brush border of intestinal epithelial cells consists of an array of tightly packed microvilli. Within each microvillus is a bundle of 20-30 actin filaments. The basal ends of the filament bundles are embedded in and interconected by a filamentous meshwork, the terminal web, which lies directly beneath the microvilli. When calcium and ATP are added to isolated brush borders that have been treated with the detergent, Triton X-100, the microvillar filament bundles rapidly retract into and through the terminal web region. Biochemical studies of brush border contractile proteins suggest that the observed microvillar contraction is actomyosin mediated. We have shown previously that the major protein of the brush border's actin (Tilney, L. G., and M. S. Mooseker. 1971. Proc. Natl. Acad. Sci. U. S. A. 68:2611-2615). The brush border also contains a protein with the same molecular weight as the heavy chain subunit of myosin (200, 000 daltons). In addition, preparations of demembranated brush borders exhibit potassium-EDTA ATPase activity of 0.02 mumol phosphate/mg-min (22 degrees C); this assay is diagnostic for myosin-like ATPase isolated from vertebrate sources. Other proteins of the brush border include a 30,000 dalton protein with properties similar to those of tropomyosin, and a protein with the same molecular weight as the Z band protein, alpha-actinin (95,000 daltons). How these observations bear on the basis for microvillar movements in vivo is discussed within the framework of our recent model for the organization of actin and myosin in the brush border (Mooseker, M. S., and L. G. Tilney. 1975. J. Cell Biol. 67:725-743). | Brush border motility. Microvillar contraction in triton-treated brush borders isolated from intestinal epithelium. The brush border of intestinal epithelial cells consists of an array of tightly packed microvilli. Within each microvillus is a bundle of 20-30 actin filaments. The basal ends of the filament bundles are embedded in and interconected by a filamentous meshwork, the terminal web, which lies directly beneath the microvilli. When calcium and ATP are added to isolated brush borders that have been treated with the detergent, Triton X-100, the microvillar filament bundles rapidly retract into and through the terminal web region. Biochemical studies of brush border contractile proteins suggest that the observed microvillar contraction is actomyosin mediated. We have shown previously that the major protein of the brush border's actin (Tilney, L. G., and M. S. Mooseker. 1971. Proc. Natl. Acad. Sci. U. S. A. 68:2611-2615). The brush border also contains a protein with the same molecular weight as the heavy chain subunit of myosin (200, 000 daltons). In addition, preparations of demembranated brush borders exhibit potassium-EDTA ATPase activity of 0.02 mumol phosphate/mg-min (22 degrees C); this assay is diagnostic for myosin-like ATPase isolated from vertebrate sources. Other proteins of the brush border include a 30,000 dalton protein with properties similar to those of tropomyosin, and a protein with the same molecular weight as the Z band protein, alpha-actinin (95,000 daltons). How these observations bear on the basis for microvillar movements in vivo is discussed within the framework of our recent model for the organization of actin and myosin in the brush border (Mooseker, M. S., and L. G. Tilney. 1975. J. Cell Biol. 67:725-743). | [
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|
PMID:11223 | pH-dependent binding of immunoglobulins to intestinal cells of the neonatal rat. | Rat and rabbit IgG immunoglobulins conjugated to horseradiah peroxidase as a histochemical marker bind at 0 degrees C to the luminal surface of absorptive cells in isolated segments of jejunum from 10-12-day old rats. Binding is observed at pH 6.0, near the normal luminal pH of the duodenum and jejunum at this age, but not at pH 7.4. Furthermore, no binding occurs when cells are exposed at pH 6.0 to either free peroxidase or peroxidase conjugated to chicken or sheep IgG immunoglobulins or bovine serum albumin. The sensitivity of binding to pH suggests a means whereby immunoglobulins which are selectively absorbed by the cells can be released efficiently at the abluminal surface. | pH-dependent binding of immunoglobulins to intestinal cells of the neonatal rat. Rat and rabbit IgG immunoglobulins conjugated to horseradiah peroxidase as a histochemical marker bind at 0 degrees C to the luminal surface of absorptive cells in isolated segments of jejunum from 10-12-day old rats. Binding is observed at pH 6.0, near the normal luminal pH of the duodenum and jejunum at this age, but not at pH 7.4. Furthermore, no binding occurs when cells are exposed at pH 6.0 to either free peroxidase or peroxidase conjugated to chicken or sheep IgG immunoglobulins or bovine serum albumin. The sensitivity of binding to pH suggests a means whereby immunoglobulins which are selectively absorbed by the cells can be released efficiently at the abluminal surface. | [
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|
PMID:11224 | Dietary phosphate deprivation in women and men: effects on mineral and acid balances, parathyroid hormone and the metabolism of 25-OH-vitamin D. | We evaluated the effects of dietary PO4 restriction on 25-OH-Vitamin D3 metabolism, serum iPTH levels, and mineral balances in healthy women and men. PO4 balances were progressively negative because of fecal losses without sex difference. Turnover of the plasma 25-OH-D pool was increased from 5.8 +/- 0.4 to 12 +/- 1.2 nmol/day; P less than 0.001, despite a fall in serum iPTH of -1.1 +/- 0.3 mulEq/ml; P less than 0.01. In both sexes, net intestinal calcium and magnesium absorption increased in proportion to a more rapid turnover of the plasma 25-OH-D pool, implying increased renal 1,25-(OH)2-D3 production. By contrast, there was a striking sex difference in the response of serum PO4 to dietary PO4 deprivation; the levels falling progressively in women, but remaining at control levels in men. Women demonstrated progressive hypercalciuria and negative Ca balances while in men the increments in intestinal Ca absorption were approximately matched by the increments in urinary Ca excretion so that Ca balances were not different from zero. | Dietary phosphate deprivation in women and men: effects on mineral and acid balances, parathyroid hormone and the metabolism of 25-OH-vitamin D. We evaluated the effects of dietary PO4 restriction on 25-OH-Vitamin D3 metabolism, serum iPTH levels, and mineral balances in healthy women and men. PO4 balances were progressively negative because of fecal losses without sex difference. Turnover of the plasma 25-OH-D pool was increased from 5.8 +/- 0.4 to 12 +/- 1.2 nmol/day; P less than 0.001, despite a fall in serum iPTH of -1.1 +/- 0.3 mulEq/ml; P less than 0.01. In both sexes, net intestinal calcium and magnesium absorption increased in proportion to a more rapid turnover of the plasma 25-OH-D pool, implying increased renal 1,25-(OH)2-D3 production. By contrast, there was a striking sex difference in the response of serum PO4 to dietary PO4 deprivation; the levels falling progressively in women, but remaining at control levels in men. Women demonstrated progressive hypercalciuria and negative Ca balances while in men the increments in intestinal Ca absorption were approximately matched by the increments in urinary Ca excretion so that Ca balances were not different from zero. | [
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|
PMID:11225 | Size heterogeneity of human serum somatomedin. | Somatomedin (SM) in unextracted human serum was studied by Sephadex chromatography, starch gel electrophoresis, and in vitro SM bioassays. Repeated rechromatography of the various serum fractions revealed SM activity in a variety of indicated molecular size (IMS) areas; greater than 90,000, (very large); 90,000-20,000, (large); 20,000-9000, (intermediate); 9000-2000, (small); and less than 2000, (very small). Chromatography of unextracted serum, at pH 7.4, revealed 65% of SM as very large SM. Most of this very large SM remained stable at neutral pH; less than 25% appeared in smaller molecular sizes after acid eluent rechromatography. The latter significantly reassociated with other serum proteins in alkaline conditions. The large IMS SM was relatively stable on rehcromatography, whereas most of the intermediate IMS SM was converted to acidic small IMS SM proteins. The small IMS SM was stable, as a basic protein, after repeated acidic rechromatography. Most of the very small IMS SM occurred in a molecular size of less than 500. These results suggest the presence of multiple molecular size forms of SM in unextracted serum. Some of the larger SM represents an SM-serum protein aggregation. The acidic SM, with a molecular size near insulin, may be derived from a larger SM. The very small SM may be a nonpolypeptide substance(s). | Size heterogeneity of human serum somatomedin. Somatomedin (SM) in unextracted human serum was studied by Sephadex chromatography, starch gel electrophoresis, and in vitro SM bioassays. Repeated rechromatography of the various serum fractions revealed SM activity in a variety of indicated molecular size (IMS) areas; greater than 90,000, (very large); 90,000-20,000, (large); 20,000-9000, (intermediate); 9000-2000, (small); and less than 2000, (very small). Chromatography of unextracted serum, at pH 7.4, revealed 65% of SM as very large SM. Most of this very large SM remained stable at neutral pH; less than 25% appeared in smaller molecular sizes after acid eluent rechromatography. The latter significantly reassociated with other serum proteins in alkaline conditions. The large IMS SM was relatively stable on rehcromatography, whereas most of the intermediate IMS SM was converted to acidic small IMS SM proteins. The small IMS SM was stable, as a basic protein, after repeated acidic rechromatography. Most of the very small IMS SM occurred in a molecular size of less than 500. These results suggest the presence of multiple molecular size forms of SM in unextracted serum. Some of the larger SM represents an SM-serum protein aggregation. The acidic SM, with a molecular size near insulin, may be derived from a larger SM. The very small SM may be a nonpolypeptide substance(s). | [
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|
PMID:11226 | Evaluation of enrichment, storage, and age of blood agar medium in relation to its ability to support growth of anaerobic bacteria. | By measuring the colony size of a variety of anaerobic bacteria isolated from clinical specimens, an evaluation was made of the benefits derived from the addition of several enrichments to blood agar medium commonly used for the growth of anaerobes. Similar methods were used to study the effects of various storage conditions and age of the medium. The results were compared with those obtained on freshly prepared and enriched blood agar plates as well as commercially available blood agar plates. Freshly prepared and enriched blood agar was found to give substantially larger colonies than could be grown on commercially obtained blood agar plates when both were inoculated and incubated under identical conditions. Storage of plating media under CO2 for periods of up to 72 h had only a minor effect on the growth of the anaerobic bacteria studied, but longer periods of storage under CO2 resulted in a less efficient plating medium. Nonenriched brain heart infusion (BHI) was found to be a better basal medium than Trypticase soy agar (TSA) medium. Colony size on fully enriched BHI blood agar plates was greater than nonenriched BHI greater than nonenriched TSA greater than commercially prepared nonenriched TSA plates. The data suggest that freshness of the plates may be as important as using rich media. | Evaluation of enrichment, storage, and age of blood agar medium in relation to its ability to support growth of anaerobic bacteria. By measuring the colony size of a variety of anaerobic bacteria isolated from clinical specimens, an evaluation was made of the benefits derived from the addition of several enrichments to blood agar medium commonly used for the growth of anaerobes. Similar methods were used to study the effects of various storage conditions and age of the medium. The results were compared with those obtained on freshly prepared and enriched blood agar plates as well as commercially available blood agar plates. Freshly prepared and enriched blood agar was found to give substantially larger colonies than could be grown on commercially obtained blood agar plates when both were inoculated and incubated under identical conditions. Storage of plating media under CO2 for periods of up to 72 h had only a minor effect on the growth of the anaerobic bacteria studied, but longer periods of storage under CO2 resulted in a less efficient plating medium. Nonenriched brain heart infusion (BHI) was found to be a better basal medium than Trypticase soy agar (TSA) medium. Colony size on fully enriched BHI blood agar plates was greater than nonenriched BHI greater than nonenriched TSA greater than commercially prepared nonenriched TSA plates. The data suggest that freshness of the plates may be as important as using rich media. | [
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|
PMID:11227 | Mycobactericidal activity of glutaraldehyde solutions. | Aqueous solutions of alkaline glutaraldehyde (buffered at pH 8.5) inactivated a standard suspension of Mycobacterium tuberculosis H37Rv faster than the corresponding acid (pH 3.7 preparation. Quantitative differences in the rate of inactivation of eight other species of Mycobacterium were determined using a 1% solution of alkaline glutaraldehyde and inactivation of residual glutaraldehyde with 1% sodium bisulfite solution. Variations in the rate of kill were observed between the various mycobacterial species tested, but such differences were probably not sufficiently large to be of practical importance. A 2% alkaline glutaraldehyde solution inactivated 10(5) viable M. tuberculosis cells present on the surface of porcelain penicylinders within 5 min at 18 degrees C. This rate of inactivation was faster than in the acidic solution. | Mycobactericidal activity of glutaraldehyde solutions. Aqueous solutions of alkaline glutaraldehyde (buffered at pH 8.5) inactivated a standard suspension of Mycobacterium tuberculosis H37Rv faster than the corresponding acid (pH 3.7 preparation. Quantitative differences in the rate of inactivation of eight other species of Mycobacterium were determined using a 1% solution of alkaline glutaraldehyde and inactivation of residual glutaraldehyde with 1% sodium bisulfite solution. Variations in the rate of kill were observed between the various mycobacterial species tested, but such differences were probably not sufficiently large to be of practical importance. A 2% alkaline glutaraldehyde solution inactivated 10(5) viable M. tuberculosis cells present on the surface of porcelain penicylinders within 5 min at 18 degrees C. This rate of inactivation was faster than in the acidic solution. | [
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|
PMID:11233 | Coordination of classroom and clinical experience. | Clinical experience, providing opportunity to apply concepts and theories presented in the classroom, is essential in the education of a dietetic practitioner--whether it be as assistant, technician, or professional. Coordination of theory and practice is needed to permit students to acquire the knowledge, attitudes, and skills necessary for competent practice. Accountable program planners and instructors are responsible for presenting curricula designed to provide such coordinated classroom and clinical experiences in a relevant, holistic approach to the education of future dietetic practitioners. | Coordination of classroom and clinical experience. Clinical experience, providing opportunity to apply concepts and theories presented in the classroom, is essential in the education of a dietetic practitioner--whether it be as assistant, technician, or professional. Coordination of theory and practice is needed to permit students to acquire the knowledge, attitudes, and skills necessary for competent practice. Accountable program planners and instructors are responsible for presenting curricula designed to provide such coordinated classroom and clinical experiences in a relevant, holistic approach to the education of future dietetic practitioners. | [
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|
PMID:11260 | Interaction of erythrocytes with human serum proteins. I. Analysis of the effect of pH and ionic strength of the medium. | The effect of ionic parameters of the medium (pH and ionic strength) on the processes of interaction of tannin-treated erythrocytes and the protein fractions of human serum (macroglobulins, microbulins and albumin) was studied in factorial experiments. Complex effect of these parametres on the processes under investigation and optimum conditions of erythrocyte sensitization were established. Subsequent fixation of antibodies by the erythrocyte diagnostic and their agglutinating activity are manifested in different mannera depending on the conditions of preceding sensitization. Important peculairities were discovered in the mechanism of interaction between the erythrocytes and various serum proteins. The obtained results should be taken into account in the production of erythrocyte antigen and antibody diagnosticums. | Interaction of erythrocytes with human serum proteins. I. Analysis of the effect of pH and ionic strength of the medium. The effect of ionic parameters of the medium (pH and ionic strength) on the processes of interaction of tannin-treated erythrocytes and the protein fractions of human serum (macroglobulins, microbulins and albumin) was studied in factorial experiments. Complex effect of these parametres on the processes under investigation and optimum conditions of erythrocyte sensitization were established. Subsequent fixation of antibodies by the erythrocyte diagnostic and their agglutinating activity are manifested in different mannera depending on the conditions of preceding sensitization. Important peculairities were discovered in the mechanism of interaction between the erythrocytes and various serum proteins. The obtained results should be taken into account in the production of erythrocyte antigen and antibody diagnosticums. | [
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PMID:11261 | Tuberculin-sensitized lymphocytes detected by altered electrophorectic mobility distributions after incubation with the antigen PPD. | Lymphocytes from donors who had had tuberculosis (a disease known to provoke a cell-mediated immunity) were incubated with the tuberculin antigen, purified protein derivative (PPD), and their distribution of electrophoretic mobility was determined by laser Doppler spectroscopy. In 75% of the cases, the distribution showed a new, high mobility cell subpopulation that was not present before exposure to the PPD. Control lymphocytes from individuals with negative skin tests or no record of tuberculosis showed no mobility changes after incubation with PPD. These observations indicate that the new mobility subpopulation arose from a specific interaction between the antigen and sensitized cells of the donors. | Tuberculin-sensitized lymphocytes detected by altered electrophorectic mobility distributions after incubation with the antigen PPD. Lymphocytes from donors who had had tuberculosis (a disease known to provoke a cell-mediated immunity) were incubated with the tuberculin antigen, purified protein derivative (PPD), and their distribution of electrophoretic mobility was determined by laser Doppler spectroscopy. In 75% of the cases, the distribution showed a new, high mobility cell subpopulation that was not present before exposure to the PPD. Control lymphocytes from individuals with negative skin tests or no record of tuberculosis showed no mobility changes after incubation with PPD. These observations indicate that the new mobility subpopulation arose from a specific interaction between the antigen and sensitized cells of the donors. | [
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PMID:11264 | Acyl specificity in triglyceride synthesis by lactating rat mammary gland. | We have studied the specificity of the acyl-CoA:diglyceride acyltransferase reaction in lactating rat mammary gland to provide a rational explanation at the enzyme level for the nonrandom distribution of fatty acids in milk fat triglycerides. Acyl-CoA:diglyceride acyltransferase activity was measured using various diglyceride and radioactive acyl-CoA substrates; products were identified as triglycerides by thin-layer and gas-liquid chromatography. Most of the enzymatic activity was located in the microsomal fraction and showed a broad specificity for the acyl donors tested C10, C12, C14, C16, C18, and C18:1 CoA esters). The acyltransferase activity was highly specific for sn-1,2-diglyceride enantiomers; rac-1,3- and sn-2,3-diglycerides were relatively inactive. The acyl-CoA specificity was not affected by the type of 1,2-diglyceride acceptor offered, although dilaurin was the best acceptor and sn-1,2-dilaurin greater than sn-1,2-dimyristin greater than sn-1,2-dipalmitin greater than sn-1,2-distearin. We have previously shown that in the microsomal fraction from lactating rat mammary gland, the acyltransferase activities concerned with the conversion of sn-glycero-3-phosphate to diacylglycerophosphate show a very marked specificity for long chain acyl-CoA's. Therefore, we conclude that the predominant localization of long chain fatty acids in the 1 and 2 positions, and of shorter chain fatty acids in the 3 position of the glycerol backbone, results at least in part from the specificities of the mammary gland acyltransferases. | Acyl specificity in triglyceride synthesis by lactating rat mammary gland. We have studied the specificity of the acyl-CoA:diglyceride acyltransferase reaction in lactating rat mammary gland to provide a rational explanation at the enzyme level for the nonrandom distribution of fatty acids in milk fat triglycerides. Acyl-CoA:diglyceride acyltransferase activity was measured using various diglyceride and radioactive acyl-CoA substrates; products were identified as triglycerides by thin-layer and gas-liquid chromatography. Most of the enzymatic activity was located in the microsomal fraction and showed a broad specificity for the acyl donors tested C10, C12, C14, C16, C18, and C18:1 CoA esters). The acyltransferase activity was highly specific for sn-1,2-diglyceride enantiomers; rac-1,3- and sn-2,3-diglycerides were relatively inactive. The acyl-CoA specificity was not affected by the type of 1,2-diglyceride acceptor offered, although dilaurin was the best acceptor and sn-1,2-dilaurin greater than sn-1,2-dimyristin greater than sn-1,2-dipalmitin greater than sn-1,2-distearin. We have previously shown that in the microsomal fraction from lactating rat mammary gland, the acyltransferase activities concerned with the conversion of sn-glycero-3-phosphate to diacylglycerophosphate show a very marked specificity for long chain acyl-CoA's. Therefore, we conclude that the predominant localization of long chain fatty acids in the 1 and 2 positions, and of shorter chain fatty acids in the 3 position of the glycerol backbone, results at least in part from the specificities of the mammary gland acyltransferases. | [
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PMID:11265 | The coordinate roles of branchial nerve activity and potassium in the stimulation of ciliary activity in Mytilus edulis: observations with phenoxybenzamine, bromolysergic acid and fluorescence histochemistry. | Potassium concentrations in excess of 30 mM increase the rate of beating of lateral cilia on the gill of Mytilus edulis. Cilioexcitation produced by low frequency (5 beats/s) electrical stimulation was potentiated with potassium but blocked with bromolysergic acid (a serotonergic inhibitor). Cilioinhibition produced by high frequency (50 beats/s) stimulation was decreased with potassium and phenoxybenzamine (a dopaminergic inhibitor). Phenoxybenzamine enhanced the cilioexcitation produced by potassium. Potassium doses incapable of maintaining a basal rate of beating (less than 30 mM) could increase ciliary activity if phenoxybenzamine was also added. After transection of the branchial nerve, the yellow-fluorophore (serotonergic storage) and cilioexcitatory effect of potassium gradually decrease. This study shows that the potassium effect on ciliary activity (a) increase with low frequency nerve stimulation, presumably through the release of serotonin and (b) decreases with high frequency nerve stimulation, presumably through the release of dopamine. | The coordinate roles of branchial nerve activity and potassium in the stimulation of ciliary activity in Mytilus edulis: observations with phenoxybenzamine, bromolysergic acid and fluorescence histochemistry. Potassium concentrations in excess of 30 mM increase the rate of beating of lateral cilia on the gill of Mytilus edulis. Cilioexcitation produced by low frequency (5 beats/s) electrical stimulation was potentiated with potassium but blocked with bromolysergic acid (a serotonergic inhibitor). Cilioinhibition produced by high frequency (50 beats/s) stimulation was decreased with potassium and phenoxybenzamine (a dopaminergic inhibitor). Phenoxybenzamine enhanced the cilioexcitation produced by potassium. Potassium doses incapable of maintaining a basal rate of beating (less than 30 mM) could increase ciliary activity if phenoxybenzamine was also added. After transection of the branchial nerve, the yellow-fluorophore (serotonergic storage) and cilioexcitatory effect of potassium gradually decrease. This study shows that the potassium effect on ciliary activity (a) increase with low frequency nerve stimulation, presumably through the release of serotonin and (b) decreases with high frequency nerve stimulation, presumably through the release of dopamine. | [
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|
PMID:11266 | Proline inhibition of a sea anemone alarm pheromone response. | 1. L-proline, by itself or in animal tissue extracts, inhibits the response of the sea anemone Anthopleura elegantissima to the alarm phermone, anthopeurine. 2. The effect of proline is mediated by a receptor that is specific for the structure and configuration of the part of the L-proline molecule containing the carboxyl and imino groups. 3. Proline inhibition is competitive, in the sense that the effects of a given proline concentration can be overridden by an increase in anthopeurine concentration. 4. The magnitude of proline inhibition increases with proline concentration and decreases as the duration of exposure to proline increases. 5. Neither the final conducting system mediating the alarm response nor the responding muscles are inhbited by proline. Inhibition presumably occurs at or soon after the level of anthopleurine receptors. 6. Proline inhibition may resolve the potential conflict between Anthopleura's mutually exclusive feeding and alarm pheromone responses. | Proline inhibition of a sea anemone alarm pheromone response. 1. L-proline, by itself or in animal tissue extracts, inhibits the response of the sea anemone Anthopleura elegantissima to the alarm phermone, anthopeurine. 2. The effect of proline is mediated by a receptor that is specific for the structure and configuration of the part of the L-proline molecule containing the carboxyl and imino groups. 3. Proline inhibition is competitive, in the sense that the effects of a given proline concentration can be overridden by an increase in anthopeurine concentration. 4. The magnitude of proline inhibition increases with proline concentration and decreases as the duration of exposure to proline increases. 5. Neither the final conducting system mediating the alarm response nor the responding muscles are inhbited by proline. Inhibition presumably occurs at or soon after the level of anthopleurine receptors. 6. Proline inhibition may resolve the potential conflict between Anthopleura's mutually exclusive feeding and alarm pheromone responses. | [
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|
PMID:11267 | An electrolytic method for determining oxygen dissociation curves using small blood samples: the effect of temperature on trout and human blood. | 1. A detailed account is given of an electrolytic method for determining the oxygen dissociation curve of fish blood using a single sample of 50-100 mul for the whole curve. The accuracy and some of the problems arising from its uses are discussed. 2. Oxygen dissociation curves have been determined for trout blood and human blood at temperatures of 15 and 37 degrees C. The relationship between P50 and temperature is similar to that obtained using other methods. Absolute values of P50 are generally lower than those obtained by other methods, especially in the case of fish blood. 3. The effect of PCO2 and pH on the oxygen dissociation curve of trout blood is tested and it is shown that PCO2 has a more marked effect than pH when the other factor is maintained at a constant level. The Bohr factor (delta log P50/delta pH) appears to be approximately the same and independent of the PCO2. 4. The P50 of ray blood determined from fish during and after an operation showed an increased Bohr factor. | An electrolytic method for determining oxygen dissociation curves using small blood samples: the effect of temperature on trout and human blood. 1. A detailed account is given of an electrolytic method for determining the oxygen dissociation curve of fish blood using a single sample of 50-100 mul for the whole curve. The accuracy and some of the problems arising from its uses are discussed. 2. Oxygen dissociation curves have been determined for trout blood and human blood at temperatures of 15 and 37 degrees C. The relationship between P50 and temperature is similar to that obtained using other methods. Absolute values of P50 are generally lower than those obtained by other methods, especially in the case of fish blood. 3. The effect of PCO2 and pH on the oxygen dissociation curve of trout blood is tested and it is shown that PCO2 has a more marked effect than pH when the other factor is maintained at a constant level. The Bohr factor (delta log P50/delta pH) appears to be approximately the same and independent of the PCO2. 4. The P50 of ray blood determined from fish during and after an operation showed an increased Bohr factor. | [
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|
PMID:11268 | Physiology of an ATP receptor in labellar sensilla of the tsetse fly Glossina morsitans morsitans Westw. (Diptera: Glossinidae). | Electrophysiological recordings have been made from cells in the eight large, labellar sensilla of g. morsitans. One of these cells in each sensillum was shown to respond to ATP over a concentration range of 10(-6)-10(-3) M. It was also sensitive to several other adenosine phophates, but much less sensitive to CTP, GTP and ITP. The activity of the receptor was depressed below pH 7, and sometimes considerably increased above pH 9. These aspects the receptor's physiology support the results of behavioural studies. It is concluded that the eight receptors mediate the flies' behavioural response to ATP. | Physiology of an ATP receptor in labellar sensilla of the tsetse fly Glossina morsitans morsitans Westw. (Diptera: Glossinidae). Electrophysiological recordings have been made from cells in the eight large, labellar sensilla of g. morsitans. One of these cells in each sensillum was shown to respond to ATP over a concentration range of 10(-6)-10(-3) M. It was also sensitive to several other adenosine phophates, but much less sensitive to CTP, GTP and ITP. The activity of the receptor was depressed below pH 7, and sometimes considerably increased above pH 9. These aspects the receptor's physiology support the results of behavioural studies. It is concluded that the eight receptors mediate the flies' behavioural response to ATP. | [
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|
PMID:11269 | Nervous control of light responses in the sea anemone, Calamactis praelongus. | 1. The burrowing sea anemone, Calamactis praelongus, responds to light with local, non-nervous contractions of the column. There are also more extensive responses of the column and retractor muscles co-ordinated by nerve net pulses (NNP's) under pacemaker control. 2. NNP's occur in at least two types of bursts and in sequences which sometimes indicate a rotating site of pulse initiation. 3. Light-evoked NNP sequences can be tape recorded and used later to drive a stimulator to reproduce the original sequences in the same or different anemones, evoking muscular responses which approximate the originals. This technique separates the pacemaker-directed component of the light response from the local effects of light stimulation. 4. Isolated circular and parietal muscles contract slowly when stimulated by light or excited indirectly by NNP's. Retractor muscles are insensitive to light but produce rapid contractions when excited by closely spaced light-evoked NNP's. 5. A model for light responses is proposed which incorporates the characteristics of isolated muscles and intact anemones. | Nervous control of light responses in the sea anemone, Calamactis praelongus. 1. The burrowing sea anemone, Calamactis praelongus, responds to light with local, non-nervous contractions of the column. There are also more extensive responses of the column and retractor muscles co-ordinated by nerve net pulses (NNP's) under pacemaker control. 2. NNP's occur in at least two types of bursts and in sequences which sometimes indicate a rotating site of pulse initiation. 3. Light-evoked NNP sequences can be tape recorded and used later to drive a stimulator to reproduce the original sequences in the same or different anemones, evoking muscular responses which approximate the originals. This technique separates the pacemaker-directed component of the light response from the local effects of light stimulation. 4. Isolated circular and parietal muscles contract slowly when stimulated by light or excited indirectly by NNP's. Retractor muscles are insensitive to light but produce rapid contractions when excited by closely spaced light-evoked NNP's. 5. A model for light responses is proposed which incorporates the characteristics of isolated muscles and intact anemones. | [
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-0.056135810911655426,
0.021778255701065063,
-0.03658369183540344,
0.030645744875073433,
-0.060798823833465576,
0.017081627622246742,
0.03539662808179855,
0.05213938653469086
]
|
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