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PMID:7919 | The effect of adrenergic blocking and stimulating agents on arginine vasotocin release in birds. | The effect of adrenergic blocking and stimulating agents on arginine vasotocin (AVT) release from the neurohpophysis following osmotic stimuli was studied in hens. It was found that alpha - adrenergic blocking drug - regitine inhibits the release of AVT after the intravenous injection of hypertonic solution of NaCl. The alpha - adrenergic stimulating drug - vasoxine as well as beta - adrenergic blocking drug - propranolol and beta - stimulating drug isoprenalinsulphate had no effect on the release of AVT. | The effect of adrenergic blocking and stimulating agents on arginine vasotocin release in birds. The effect of adrenergic blocking and stimulating agents on arginine vasotocin (AVT) release from the neurohpophysis following osmotic stimuli was studied in hens. It was found that alpha - adrenergic blocking drug - regitine inhibits the release of AVT after the intravenous injection of hypertonic solution of NaCl. The alpha - adrenergic stimulating drug - vasoxine as well as beta - adrenergic blocking drug - propranolol and beta - stimulating drug isoprenalinsulphate had no effect on the release of AVT. | [
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PMID:7920 | Studies on the activity of cholinesterase included on silica gel. | Cholinesterase was immobilized in silica gel. The activity of the bound enzyme was determined by comparing the concentrations of hydrogen ions in the solution entering and leaving the reactor, filled with a suspension of the insoluble enzyme in Sephadex. | Studies on the activity of cholinesterase included on silica gel. Cholinesterase was immobilized in silica gel. The activity of the bound enzyme was determined by comparing the concentrations of hydrogen ions in the solution entering and leaving the reactor, filled with a suspension of the insoluble enzyme in Sephadex. | [
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PMID:7926 | Amino acid transport in isolated neurons and glia. | Our efforts have been directed towards characterizing amino acid uptake, metabolism and release in bulk-isolated glia and neuronal perikarya studied in parallel with nerve-endings, especially as it concerns the transmitter amino acids and the participation of glia in the clearing of the synpatic space during impulse conduction. A possible neuromodulator role for the glia at the synapse is also suggested by K+-stimulated release. Our most definitive conclusions have been based so far on studies with GABA, although we are also beginning to accumulate data for glutamate related to glutamate-glutamine compartmentation. Glia preferentially accumulate potassium and amino acids compared to neuronal perikarya, have higher Na+/K+-ATPase activity, possess high-affinity, sodium-dependent uptake systems for GABA and glutamate similar to the ones in synaptosomes, and release amino acid in response to a potassium pulse by a calcium-independent process. Low neuronal uptake could be due to loss of dendrites. Unidirectional GABA-flux from the synaptosomal to glial compartment is supported by high GAD in nerve endings compared to high GABA-T in glia. Glutamine may be a transmitter glutamate-precursor in nerve-endings since glutaminase activity is high in nerve-endings, but low in glia where glutamine is presumably made. Glutamine uptake in both glia and synaptosomes obeys low-affinity kinetics in contrast to glutamate, consistent with the inability of glutamine to excite the neuronal membrane. The studies with GABA, which are considerably more extensive, are supported by related work using glia in tissue-culture and autoradiography. There appears to be a suggested difference in the behavior of amines which were poorly taken up by the glial system. Glia, synaptosomes and neuronal perikarya, in general behaved similarly with respect to requirements for uptake and release, except in the case of Ca++, which exerted opposite effects on glial and synaptosomal uptake of GABA. We believe that work along these lines tends to firmly establish a direct role for glial cells as modulators of neuronal excitability and represents a convergence between transmitter amino acid neuropharmacology and cellular biochemistry. This not only deepens and enlarges the vocabulary of synaptic biochemistry but also undoubtedly will have major clinical applications in the fields of epilepsy and behavior. | Amino acid transport in isolated neurons and glia. Our efforts have been directed towards characterizing amino acid uptake, metabolism and release in bulk-isolated glia and neuronal perikarya studied in parallel with nerve-endings, especially as it concerns the transmitter amino acids and the participation of glia in the clearing of the synpatic space during impulse conduction. A possible neuromodulator role for the glia at the synapse is also suggested by K+-stimulated release. Our most definitive conclusions have been based so far on studies with GABA, although we are also beginning to accumulate data for glutamate related to glutamate-glutamine compartmentation. Glia preferentially accumulate potassium and amino acids compared to neuronal perikarya, have higher Na+/K+-ATPase activity, possess high-affinity, sodium-dependent uptake systems for GABA and glutamate similar to the ones in synaptosomes, and release amino acid in response to a potassium pulse by a calcium-independent process. Low neuronal uptake could be due to loss of dendrites. Unidirectional GABA-flux from the synaptosomal to glial compartment is supported by high GAD in nerve endings compared to high GABA-T in glia. Glutamine may be a transmitter glutamate-precursor in nerve-endings since glutaminase activity is high in nerve-endings, but low in glia where glutamine is presumably made. Glutamine uptake in both glia and synaptosomes obeys low-affinity kinetics in contrast to glutamate, consistent with the inability of glutamine to excite the neuronal membrane. The studies with GABA, which are considerably more extensive, are supported by related work using glia in tissue-culture and autoradiography. There appears to be a suggested difference in the behavior of amines which were poorly taken up by the glial system. Glia, synaptosomes and neuronal perikarya, in general behaved similarly with respect to requirements for uptake and release, except in the case of Ca++, which exerted opposite effects on glial and synaptosomal uptake of GABA. We believe that work along these lines tends to firmly establish a direct role for glial cells as modulators of neuronal excitability and represents a convergence between transmitter amino acid neuropharmacology and cellular biochemistry. This not only deepens and enlarges the vocabulary of synaptic biochemistry but also undoubtedly will have major clinical applications in the fields of epilepsy and behavior. | [
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PMID:7933 | Mediation of macrophage effector function by lymphokine. | The progressive effect of lymphokine contact on macrophages in vitro has been studied using various quantitative cytochemical techniques. Changes in the physiology of the macrophages have been seen rapidly after lymphokine contact. These appear to correlate with the functional effect of inhibition of migration. After more prolonged contact with lymphokine however, the macrophages exhibit different changes to their physiology and reach a state of enhanced cytochemical activity which has been termed 'activation'. By comparing the rapid effect of lymphokine to the changes seen after prolonged contact it is suggested that one can rationalize the apparent paradoxical effects of these soluble mediators which appear initially to 'turn off' the macrophage and subsequently 'activate' the same cell. In doing so an hypothesis is put forward as to how this mediator might work in vivo. | Mediation of macrophage effector function by lymphokine. The progressive effect of lymphokine contact on macrophages in vitro has been studied using various quantitative cytochemical techniques. Changes in the physiology of the macrophages have been seen rapidly after lymphokine contact. These appear to correlate with the functional effect of inhibition of migration. After more prolonged contact with lymphokine however, the macrophages exhibit different changes to their physiology and reach a state of enhanced cytochemical activity which has been termed 'activation'. By comparing the rapid effect of lymphokine to the changes seen after prolonged contact it is suggested that one can rationalize the apparent paradoxical effects of these soluble mediators which appear initially to 'turn off' the macrophage and subsequently 'activate' the same cell. In doing so an hypothesis is put forward as to how this mediator might work in vivo. | [
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PMID:7934 | Inhibition of prostaglandin synthesis in vivo by nonsteroid anti-inflammatory drugs: evidence for the importance of pharmacokinetics. | A variety of acidic and non-acidic compounds are potent inhibitors of prostaglandin (PG) synthesis in vitro. However, only a few, namely the acidic nonsteroid anti-inflammatory drugs (NSAID) are useful anti-inflammatory analgesics in the clinic. Since inhibition of PG-synthesis is believed to be the main target of NSAID in inflammation this superiority of acidic compounds remains unexplained. We have considered that one explanation could be that only acidic NSAID appear in high concentrations in inflamed tissue to inhibit PG-synthesis sufficiently. To test this hypothesis the following experiments were carried out: (A) PG-synthesis and its inhibition by acidic and non-acidic NSAID was measured in vivo at the site of inflammation. It was found that in therapeutic doses only acidic NSAID were capable to reduce PG-synthesis significantly. (B) Measurement of drug concentration in inflamed tissue showed that only acidic NSAID were found in significantly higher concentrations in inflamed than in control tissue. From these observations it is concluded that a specific pharmacokinetic behaviour of acidic NSAID leading to high concentrations in inflamed tissue is a decisive aspect of their anti-inflammatory action. | Inhibition of prostaglandin synthesis in vivo by nonsteroid anti-inflammatory drugs: evidence for the importance of pharmacokinetics. A variety of acidic and non-acidic compounds are potent inhibitors of prostaglandin (PG) synthesis in vitro. However, only a few, namely the acidic nonsteroid anti-inflammatory drugs (NSAID) are useful anti-inflammatory analgesics in the clinic. Since inhibition of PG-synthesis is believed to be the main target of NSAID in inflammation this superiority of acidic compounds remains unexplained. We have considered that one explanation could be that only acidic NSAID appear in high concentrations in inflamed tissue to inhibit PG-synthesis sufficiently. To test this hypothesis the following experiments were carried out: (A) PG-synthesis and its inhibition by acidic and non-acidic NSAID was measured in vivo at the site of inflammation. It was found that in therapeutic doses only acidic NSAID were capable to reduce PG-synthesis significantly. (B) Measurement of drug concentration in inflamed tissue showed that only acidic NSAID were found in significantly higher concentrations in inflamed than in control tissue. From these observations it is concluded that a specific pharmacokinetic behaviour of acidic NSAID leading to high concentrations in inflamed tissue is a decisive aspect of their anti-inflammatory action. | [
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PMID:7935 | Electrophoretic separation and characterization of subunits released from influenza virus by detergents. | Subunits released from influenza A/Singapore/1/57 (H2N2) virus by either Triton-X-100 (T-X-100); or sodium lauryl sarcosinate (SLS) or ether were separated by electrophoresis in agarose suspension into a rapidly migrating fraction (I) and a slowly migrating fraction (II). Fraction I obtained after T-X-100 treatment contained the viral ribonucleoprotein (RNP) in a form indistinguishable from the obtained after ether treatment. SLS treatment of the virus resulted in a rapidly migrating fraction containing only the protein part of the viral RNP. Fraction II obtained after T-X-100 or SLS treatment contained both haemagglutinin (HA) and neuraminidase (NA), mostly dissociated from each other, in contrast to fraction II obtained after ether treatment which contained mixed aggregates of HA and NA. The yields of electrophoretically isolated RNP and HA-NA were essentially the same irrespective of whether T-X-100 or ether was used for virus disruption. Treatment of virus by T-X-100 and subsequent removal of the latter resulted in a 10-20-fold increase of the HA activity. After sodium dodecyl sulphate (SDS) treatment of the virus, the NA activity was found in a heterogeneous fraction with surprisingly high migration rate towards the anode, indicating that NA remained active despite its extensive SDS binding. | Electrophoretic separation and characterization of subunits released from influenza virus by detergents. Subunits released from influenza A/Singapore/1/57 (H2N2) virus by either Triton-X-100 (T-X-100); or sodium lauryl sarcosinate (SLS) or ether were separated by electrophoresis in agarose suspension into a rapidly migrating fraction (I) and a slowly migrating fraction (II). Fraction I obtained after T-X-100 treatment contained the viral ribonucleoprotein (RNP) in a form indistinguishable from the obtained after ether treatment. SLS treatment of the virus resulted in a rapidly migrating fraction containing only the protein part of the viral RNP. Fraction II obtained after T-X-100 or SLS treatment contained both haemagglutinin (HA) and neuraminidase (NA), mostly dissociated from each other, in contrast to fraction II obtained after ether treatment which contained mixed aggregates of HA and NA. The yields of electrophoretically isolated RNP and HA-NA were essentially the same irrespective of whether T-X-100 or ether was used for virus disruption. Treatment of virus by T-X-100 and subsequent removal of the latter resulted in a 10-20-fold increase of the HA activity. After sodium dodecyl sulphate (SDS) treatment of the virus, the NA activity was found in a heterogeneous fraction with surprisingly high migration rate towards the anode, indicating that NA remained active despite its extensive SDS binding. | [
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PMID:7936 | The effect of combination of different inducers on the refractory state in interferon production. | A second injection of 100 mug poly (rI) poly (rC) per mouse at 6 and 24 hours after the first injection stimulated additional peaks of interferon production. The dynamics of the process of accumulation and disappearance of interferon was similar to that after a single injection of poly (rI). poly (rC). Injection of the above dose 12 hours after the first injection induced no interferon production as it apparently coincided with the refractory state in interferon production. After pretreatment of poly (rI) poly (rC) with DEAE-dextran, the refractory phase occurred in 6 hours. Inoculation of Venezuelan equine encephalomyelitis virus as a second interferon inducer resulted in a repeated stimulation of interferon production both in animals and in tissue culture; however, interferon titres in this case were low. The use of an inactivated virus as a second interferon inducer stimulated interferon production to higher titres (5120 IU/ml) than a single injection of DEAE-dextran-treated poly (rI). poly (rC). It is possible that a combined use of poly (rI). poly (rC) and noninfectious virus as a second interferon inducer eliminates the development of the refractory state. | The effect of combination of different inducers on the refractory state in interferon production. A second injection of 100 mug poly (rI) poly (rC) per mouse at 6 and 24 hours after the first injection stimulated additional peaks of interferon production. The dynamics of the process of accumulation and disappearance of interferon was similar to that after a single injection of poly (rI). poly (rC). Injection of the above dose 12 hours after the first injection induced no interferon production as it apparently coincided with the refractory state in interferon production. After pretreatment of poly (rI) poly (rC) with DEAE-dextran, the refractory phase occurred in 6 hours. Inoculation of Venezuelan equine encephalomyelitis virus as a second interferon inducer resulted in a repeated stimulation of interferon production both in animals and in tissue culture; however, interferon titres in this case were low. The use of an inactivated virus as a second interferon inducer stimulated interferon production to higher titres (5120 IU/ml) than a single injection of DEAE-dextran-treated poly (rI). poly (rC). It is possible that a combined use of poly (rI). poly (rC) and noninfectious virus as a second interferon inducer eliminates the development of the refractory state. | [
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PMID:7937 | The dynamics of development of cell resistance to viruses induced by synthetic polynucleotides. | The dynamics of interaction of complexes of synthetic polynucleotides (polyinosinic and polycitidylic acid--poly (rI)-poly (rC), and polyguanylic and polycytidylic acid--poly (rG)-poly (rC)) with cells as well as the dynamics of interferon accumulation and development of antiviral effect against some RNA viruses were studied in primary chick embryo cell (CEC) cultures. Four phases were observed in the development of the antiviral effect of synthetic polynucleotides: adsorption, increase, marked antiviral effect and waning. The duration and extent of the antiviral effect depended upon the activity and the dose of the preparation and less so upon virus type. At the same time, the dynamics of the development of the antiviral effect in early stages differed significantly depending on the virus model. | The dynamics of development of cell resistance to viruses induced by synthetic polynucleotides. The dynamics of interaction of complexes of synthetic polynucleotides (polyinosinic and polycitidylic acid--poly (rI)-poly (rC), and polyguanylic and polycytidylic acid--poly (rG)-poly (rC)) with cells as well as the dynamics of interferon accumulation and development of antiviral effect against some RNA viruses were studied in primary chick embryo cell (CEC) cultures. Four phases were observed in the development of the antiviral effect of synthetic polynucleotides: adsorption, increase, marked antiviral effect and waning. The duration and extent of the antiviral effect depended upon the activity and the dose of the preparation and less so upon virus type. At the same time, the dynamics of the development of the antiviral effect in early stages differed significantly depending on the virus model. | [
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PMID:7938 | Biochemical classification of herpes simplex virus types 1 and 2, and of intermediate strains on the basis of different susceptibilities of thymidine kinase to thymidine analogues. | Herpes simplex virus (HSV) type 1 can be differentiated from HSV type 2 on the basis of the sensitivity to 2'-deoxythymidine-5'-monophosphate of thymidine kinase induced in primary rabbit kidney cells. Whereas thymidine kinase induced by five strains of HSV type 1 (TK 1) is stimulated by suitable concentrations of 2'-deoxythymidine-5'-monophosphate, thymidine kinase induced by eight strains of HSV type 2 (TK 2) is inhibited. On the other hand, TK 2 is strongly inhibited by 2'-deoxythymidine-5'-triphosphate and by 2-bromo-2'-deoxyuridine-5'-triphosphate. The investigation of TK induced by six freshly isolated strains of HSV cross-reacting in neutralisation tests revealed two strains which induced TK 1 and two strains which induced TK 2. Two other strains induced thymidine kinase, the activity of which under the influence of these thymidine analogues was between that of TK 1 and TK 2. The properties of thymidine kinase remained constant after cloning the virus and thus is a genetically fixed trait due to recombination which could well occur in vivo. | Biochemical classification of herpes simplex virus types 1 and 2, and of intermediate strains on the basis of different susceptibilities of thymidine kinase to thymidine analogues. Herpes simplex virus (HSV) type 1 can be differentiated from HSV type 2 on the basis of the sensitivity to 2'-deoxythymidine-5'-monophosphate of thymidine kinase induced in primary rabbit kidney cells. Whereas thymidine kinase induced by five strains of HSV type 1 (TK 1) is stimulated by suitable concentrations of 2'-deoxythymidine-5'-monophosphate, thymidine kinase induced by eight strains of HSV type 2 (TK 2) is inhibited. On the other hand, TK 2 is strongly inhibited by 2'-deoxythymidine-5'-triphosphate and by 2-bromo-2'-deoxyuridine-5'-triphosphate. The investigation of TK induced by six freshly isolated strains of HSV cross-reacting in neutralisation tests revealed two strains which induced TK 1 and two strains which induced TK 2. Two other strains induced thymidine kinase, the activity of which under the influence of these thymidine analogues was between that of TK 1 and TK 2. The properties of thymidine kinase remained constant after cloning the virus and thus is a genetically fixed trait due to recombination which could well occur in vivo. | [
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PMID:7939 | Persistent infection of BHK cells with herpes simplex virus types 1 and 2 in the absence of specific anti-herpetic antibody. | Persistent infection of BHK-21 cells with 5 strains of herpes simplex virus (HSV) type 1 and a 1 strain of HSV type 2 is described. Acute infection only was obtained with two strains of type 1 and one strain of type 2. Persistent infection could not be established in HeLa and rabbit lung cells. Persistent infection of BHK cells proceeded in the absence of specific antibody. Persistently infected cells were more resistant to superinfection with homologous or heterologous HSV as compared to infection of normal BHK cells. No interferon activity was demonstrated in persistently infected cell cultures. The results of virus titration were in accordance with the demonstration of fluorescent antigen of HSV. | Persistent infection of BHK cells with herpes simplex virus types 1 and 2 in the absence of specific anti-herpetic antibody. Persistent infection of BHK-21 cells with 5 strains of herpes simplex virus (HSV) type 1 and a 1 strain of HSV type 2 is described. Acute infection only was obtained with two strains of type 1 and one strain of type 2. Persistent infection could not be established in HeLa and rabbit lung cells. Persistent infection of BHK cells proceeded in the absence of specific antibody. Persistently infected cells were more resistant to superinfection with homologous or heterologous HSV as compared to infection of normal BHK cells. No interferon activity was demonstrated in persistently infected cell cultures. The results of virus titration were in accordance with the demonstration of fluorescent antigen of HSV. | [
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PMID:7940 | Reactions of immune sera against the nucleocapsid, envelope and whole herpes simplex virus type 1. | Antibodies against (a) naked particles, (b) virus envelope, and (c) whole herpes simplex virus (HSV) type 1 were investigated. Immune rabbit serum against naked particles (and formalized naked particles) contained virus neutralizing (VN) antibody in a low titre as compared to the titre of complement-fixing (CF) antibody. Immune rabbit serum against the viral envelope had similar titres of VN and CF antibodies. In gel double diffusion precipitation tests, whole HSV gave two precipitation zones with antibodies against naked particles two zones with antibodies against the virus envelope, several zones with antibodies against whole virus and two marked zones against human convalescent serum. Viral nucleocapsid reacted with two zones with antibodies against the nucleocapsid and a faint zone with antibodies against the virus envelope. The envelopes gave two zones with antibodies against whole virus and a faint zone with antibodies against naked particles. Human convalescent serum gave a single precipitation zone with viral nucleocapsid and two zones with virus envelope and whole virus. | Reactions of immune sera against the nucleocapsid, envelope and whole herpes simplex virus type 1. Antibodies against (a) naked particles, (b) virus envelope, and (c) whole herpes simplex virus (HSV) type 1 were investigated. Immune rabbit serum against naked particles (and formalized naked particles) contained virus neutralizing (VN) antibody in a low titre as compared to the titre of complement-fixing (CF) antibody. Immune rabbit serum against the viral envelope had similar titres of VN and CF antibodies. In gel double diffusion precipitation tests, whole HSV gave two precipitation zones with antibodies against naked particles two zones with antibodies against the virus envelope, several zones with antibodies against whole virus and two marked zones against human convalescent serum. Viral nucleocapsid reacted with two zones with antibodies against the nucleocapsid and a faint zone with antibodies against the virus envelope. The envelopes gave two zones with antibodies against whole virus and a faint zone with antibodies against naked particles. Human convalescent serum gave a single precipitation zone with viral nucleocapsid and two zones with virus envelope and whole virus. | [
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PMID:7941 | Gel chromatography of serum from mice infected with a virus elevating L-lactate: NAD oxidoreductase activity--an attempt to separate viral and enzymatic activities. | Gel chromatography on Spheron P-500 of mouse serum yielded fractions elevating lactate dehydrogenase activity. The order of elution was viral activity, proteins and lactate dehydrogenase activity. Viral contamination occurred also in certain fractions subjected to repeated gel chromatography in which no proteins were detectable. | Gel chromatography of serum from mice infected with a virus elevating L-lactate: NAD oxidoreductase activity--an attempt to separate viral and enzymatic activities. Gel chromatography on Spheron P-500 of mouse serum yielded fractions elevating lactate dehydrogenase activity. The order of elution was viral activity, proteins and lactate dehydrogenase activity. Viral contamination occurred also in certain fractions subjected to repeated gel chromatography in which no proteins were detectable. | [
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PMID:7942 | Further experiments on the action of antithymocyte serum in experimental Junín virus infection. | Rabbit anti-mouse thymocyte serum (ATS) administrated as late as 7 days after infection suppressed host cell-mediated responsiveness to intracerebrally injected Junín virus, thereby diminishing the morbility and mortality of this infection. It did not affect either the humoral antibody response or the virus titer in brain. This findings suggest that: a) in mouse brain cells, Junín virus infection is basically non-cytopathic: b) cell-mediated immunity is responsible for morbility and mortality and does not clear up virus from brain as in other viral encephalitides; and c) ATS may be of therapeutic interest by suppressing or diminishing the cell-mediated response to Junín virus. | Further experiments on the action of antithymocyte serum in experimental Junín virus infection. Rabbit anti-mouse thymocyte serum (ATS) administrated as late as 7 days after infection suppressed host cell-mediated responsiveness to intracerebrally injected Junín virus, thereby diminishing the morbility and mortality of this infection. It did not affect either the humoral antibody response or the virus titer in brain. This findings suggest that: a) in mouse brain cells, Junín virus infection is basically non-cytopathic: b) cell-mediated immunity is responsible for morbility and mortality and does not clear up virus from brain as in other viral encephalitides; and c) ATS may be of therapeutic interest by suppressing or diminishing the cell-mediated response to Junín virus. | [
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PMID:7943 | The value of complement fixation and haemagglutination inhibition tests in the diagnosis of influenza A. | Antibody response of 133 influenza A patients from three outbreaks since 1972-73 was analyzed by complement fixation (CF) and haemagglutination inhibition (HI) methods. During the first outbreak, a significant response was more often measured by CF than by HI. During the last outbreak HI appeared more useful than CF for routine serological diagnosis; 23% of cases verified by HI were missed by CF. The poor response of CF antibodies was associated with the high level of pre-infection antibodies. | The value of complement fixation and haemagglutination inhibition tests in the diagnosis of influenza A. Antibody response of 133 influenza A patients from three outbreaks since 1972-73 was analyzed by complement fixation (CF) and haemagglutination inhibition (HI) methods. During the first outbreak, a significant response was more often measured by CF than by HI. During the last outbreak HI appeared more useful than CF for routine serological diagnosis; 23% of cases verified by HI were missed by CF. The poor response of CF antibodies was associated with the high level of pre-infection antibodies. | [
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PMID:7944 | Susceptibility of various cell lines to virulent and attenuated strains of pseudorabies virus. | Various cell lines were infected with virulent and attenuated strains of pseudorabies virus (PRV). According to the type of the cytopathic effect (CPE), the cells could be divided into 3 groups: cells in which all strains formed syncytia; cells in which all strains caused rounding of cells; and cells in which virulent strains caused syncytium formation while attenuated ones rounding of cells. Neither the form (fibroblastoid or epithelial) nor the origin of cells influenced the type of the CPE. L cells and some cell lines derived from human tissue proved to be less sensitive to PRV than other cells. In certain human cells (e.g. HeLa and D6) virulent PRV strains propagated better than attenuated ones. | Susceptibility of various cell lines to virulent and attenuated strains of pseudorabies virus. Various cell lines were infected with virulent and attenuated strains of pseudorabies virus (PRV). According to the type of the cytopathic effect (CPE), the cells could be divided into 3 groups: cells in which all strains formed syncytia; cells in which all strains caused rounding of cells; and cells in which virulent strains caused syncytium formation while attenuated ones rounding of cells. Neither the form (fibroblastoid or epithelial) nor the origin of cells influenced the type of the CPE. L cells and some cell lines derived from human tissue proved to be less sensitive to PRV than other cells. In certain human cells (e.g. HeLa and D6) virulent PRV strains propagated better than attenuated ones. | [
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PMID:7946 | Incidence of arbovirus antibodies in bovine, ovine and human sera collected in Eastern Sicily. | The incidence of antibodies against 9 arboviruses in ovine, bovine and human sera collected in Eastern Sicily was studied. A high incidence of antibodies was found especially against Bhanja virus in all sera and against Sandfly fever Sicilian in ovine sera only. The incidence was higher than that reported in 1971 for the western part of Sicily. These findings suggest an increase in the circulation of arboviruses in this area; the causes are briefly discussed. | Incidence of arbovirus antibodies in bovine, ovine and human sera collected in Eastern Sicily. The incidence of antibodies against 9 arboviruses in ovine, bovine and human sera collected in Eastern Sicily was studied. A high incidence of antibodies was found especially against Bhanja virus in all sera and against Sandfly fever Sicilian in ovine sera only. The incidence was higher than that reported in 1971 for the western part of Sicily. These findings suggest an increase in the circulation of arboviruses in this area; the causes are briefly discussed. | [
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PMID:7947 | "White-wild" (variola-like) poxvirus strains from rodents in Equatorial Africa. | Strains of variola-like virus were isolated from rodents, namely 1 each from Mastomys coucha (fam. Muridae) and Helioscorus rufobrachium (fam. Sciuridae) in the Bumba zone of the Equatorial province of the Zaire Republic. Attempts to isolate the virus from 48 monkeys were negative. Neutralizing antibodies were found in 1 of 7 monkey (Cercopithecus ascanius schmidti) sera examined as well as in the serum of H. rufobrachium from which the virus was isolated. | "White-wild" (variola-like) poxvirus strains from rodents in Equatorial Africa. Strains of variola-like virus were isolated from rodents, namely 1 each from Mastomys coucha (fam. Muridae) and Helioscorus rufobrachium (fam. Sciuridae) in the Bumba zone of the Equatorial province of the Zaire Republic. Attempts to isolate the virus from 48 monkeys were negative. Neutralizing antibodies were found in 1 of 7 monkey (Cercopithecus ascanius schmidti) sera examined as well as in the serum of H. rufobrachium from which the virus was isolated. | [
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PMID:7948 | Variable response of normal and transformed mouse cells to interferon inducers. | Balb/3T3 mouse cells, normal and SV40-transformed, produced interferon when induced with Newcastle disease and influenza viruses; transformed cells failed to respond to double-stranded RNA inducers. | Variable response of normal and transformed mouse cells to interferon inducers. Balb/3T3 mouse cells, normal and SV40-transformed, produced interferon when induced with Newcastle disease and influenza viruses; transformed cells failed to respond to double-stranded RNA inducers. | [
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PMID:7949 | Consumption of complement in chelated and nonchelated guinea pig serum after challenge with some viral and nonviral interferon inducers. | The consumption of complement observed during the induction of interferon by various inducers may proceed via the alternate pathway. The alternate pathway requires the presence of Mg ions in the medium while Ca ions may be absent. | Consumption of complement in chelated and nonchelated guinea pig serum after challenge with some viral and nonviral interferon inducers. The consumption of complement observed during the induction of interferon by various inducers may proceed via the alternate pathway. The alternate pathway requires the presence of Mg ions in the medium while Ca ions may be absent. | [
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PMID:7951 | The dynamics of production and the sensitivity to cycloheximide and actinomycin D of interferon-inducing and interferon messenger RNA. | In the process of virus-induced interferon production, two kinds of RNA appear in the cells. One of them induces production by the recipient cells of interferon with the species-specificity of the latter cells (interferon-inducing RNA), and the other is translated by the recipient cells pre-treated with actinomycin D into interferon with the species-specificity of the donor cells of RNA (interferon mRNA). The interferon-inducing RNA appears 20-30 minutes after virus induction and shows maximal activity after 1 hour. Its formation is not influenced by cycloheximide or actinomycin D. This RNA is assumed to be a transcriptive intermediate form of viral RNA. Interferon mRNA appears in the cells 1 hour after virus induction and shows maximal activity after 6-8 hours. Its synthesis is inhibited by cycloheximide and actinomycin D. | The dynamics of production and the sensitivity to cycloheximide and actinomycin D of interferon-inducing and interferon messenger RNA. In the process of virus-induced interferon production, two kinds of RNA appear in the cells. One of them induces production by the recipient cells of interferon with the species-specificity of the latter cells (interferon-inducing RNA), and the other is translated by the recipient cells pre-treated with actinomycin D into interferon with the species-specificity of the donor cells of RNA (interferon mRNA). The interferon-inducing RNA appears 20-30 minutes after virus induction and shows maximal activity after 1 hour. Its formation is not influenced by cycloheximide or actinomycin D. This RNA is assumed to be a transcriptive intermediate form of viral RNA. Interferon mRNA appears in the cells 1 hour after virus induction and shows maximal activity after 6-8 hours. Its synthesis is inhibited by cycloheximide and actinomycin D. | [
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PMID:7952 | Efficacy of cardioselective beta adrenergic blockade with intravenously administered tolamolol in the treatment of cardiac arrhythmias. | The efficacy of tolamolol, a cardioselective beta adrenergic blocking agent, was evaluated in the treatment of cardiac arrhythmias in 27 patients. Nineteen patients had supraventricular arrhythmias and eight had ventricular arrhythmias. Evaluation was by doulbe-blind randomized trial in 23 patients. Tolamolol was effective in reducing ventricular rate in 17 (85 percent) of 19 patients with supraventricular arrhythmias and resulted in conversion to sinus rhythm in 2 of the 17. The mean ventricular rate in 17 patients decreased from 135 to 102/min 10 minutes after initiation of administration of tolamolol and gradually decreased further to 93/min after 60 minutes. Reduction in ventricular rate was sustained for 2 hours of monitoring undergone by all patients and for 4 and 6 hours monitoring in two subgroups. Among the eight patients with ventricular ectopic beats, tolamolol reduced their frequency in four patients and had no effect in four. Six patients had chronic obstructive pulmonary disease and experienced no adverse clinical effects on respiratory function in association with administration to tolamolol. Untoward effects occurred in 10 patients, including hypotension in 3, 1 of whom required vasopressor therapy. Other side effects were sedation, nausea, dyspnea and warmth in the chest, all of which were mild and transient, requiring no treatment. Cardioselective beta adrenergic blockade with tolamolol was highly effective in controlling ventricular rate in supraventricular arrhythmias and reduced the frequency of ventricular ectopic beats in half of the small group of patients with this arrhythmia. It is particularly applicable in patients with obstructive pulmonary disease in whom cardiac beta adrenergic blockade is indicated. Hypotension is an important potential side effect. | Efficacy of cardioselective beta adrenergic blockade with intravenously administered tolamolol in the treatment of cardiac arrhythmias. The efficacy of tolamolol, a cardioselective beta adrenergic blocking agent, was evaluated in the treatment of cardiac arrhythmias in 27 patients. Nineteen patients had supraventricular arrhythmias and eight had ventricular arrhythmias. Evaluation was by doulbe-blind randomized trial in 23 patients. Tolamolol was effective in reducing ventricular rate in 17 (85 percent) of 19 patients with supraventricular arrhythmias and resulted in conversion to sinus rhythm in 2 of the 17. The mean ventricular rate in 17 patients decreased from 135 to 102/min 10 minutes after initiation of administration of tolamolol and gradually decreased further to 93/min after 60 minutes. Reduction in ventricular rate was sustained for 2 hours of monitoring undergone by all patients and for 4 and 6 hours monitoring in two subgroups. Among the eight patients with ventricular ectopic beats, tolamolol reduced their frequency in four patients and had no effect in four. Six patients had chronic obstructive pulmonary disease and experienced no adverse clinical effects on respiratory function in association with administration to tolamolol. Untoward effects occurred in 10 patients, including hypotension in 3, 1 of whom required vasopressor therapy. Other side effects were sedation, nausea, dyspnea and warmth in the chest, all of which were mild and transient, requiring no treatment. Cardioselective beta adrenergic blockade with tolamolol was highly effective in controlling ventricular rate in supraventricular arrhythmias and reduced the frequency of ventricular ectopic beats in half of the small group of patients with this arrhythmia. It is particularly applicable in patients with obstructive pulmonary disease in whom cardiac beta adrenergic blockade is indicated. Hypotension is an important potential side effect. | [
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PMID:7953 | Ultrastructural changes of the canine gastric mucosa after topical application of graded concentrations of ethanol. | Changes in the fine structure of the gastric mucosa following exposure to graded concentrations of ethanol were studied in dogs. 300 ml of 12.5, 20, and 40%, vol/vol, were instilled intragastrically for 30 min. Mucosa from the midbody and midantrum along the greater curvature was examined by light and electron microscopy. Ethanol produced a gradation of changes in the surface epithelial cells and in the lamina propria without affecting the parietal cells and chief cells. 12.5% ethanol produced widened and irregular intercellular spaces while 20 and 40% disrupted the apical cell membrane with concomitant exudation of mucin into the gastric lumen. These changes were more severe after 40% ethanol. The tight junction between cells remained intact following exposure to the lower concentrations of ethanol, but focal separation of cell junctions was observed in severely damaged areas. Quantitation of protein, sodium, and potassium concentrations in the gastric contents revealed marked increases following exposure to ethanol which correlated with the concentration. These studies provide additional morphological data on the relationship between structural changes and functional abnormalities induced by agents which break the gastric mucosal barrier. | Ultrastructural changes of the canine gastric mucosa after topical application of graded concentrations of ethanol. Changes in the fine structure of the gastric mucosa following exposure to graded concentrations of ethanol were studied in dogs. 300 ml of 12.5, 20, and 40%, vol/vol, were instilled intragastrically for 30 min. Mucosa from the midbody and midantrum along the greater curvature was examined by light and electron microscopy. Ethanol produced a gradation of changes in the surface epithelial cells and in the lamina propria without affecting the parietal cells and chief cells. 12.5% ethanol produced widened and irregular intercellular spaces while 20 and 40% disrupted the apical cell membrane with concomitant exudation of mucin into the gastric lumen. These changes were more severe after 40% ethanol. The tight junction between cells remained intact following exposure to the lower concentrations of ethanol, but focal separation of cell junctions was observed in severely damaged areas. Quantitation of protein, sodium, and potassium concentrations in the gastric contents revealed marked increases following exposure to ethanol which correlated with the concentration. These studies provide additional morphological data on the relationship between structural changes and functional abnormalities induced by agents which break the gastric mucosal barrier. | [
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PMID:7954 | Some minimal effects of bile acid on canine gastric mucosa in Heidenhain pouches. | The gastric mucosa in four canine Heidenhain pouches was damaged with sodium taurocholate in a concentration which was just sufficient in each dog to produce consistent changes. Solutions were placed in the pouches and changes in the volume, concentrations, and fluxes of both hydrogen ion and sodium ion were measured before and during exposure to taurocholate. Changes which occurred with taurocholate were compared with control periods; taurocholate caused a relative increase in volume of 1 ml and approximately a fourfold increase in the hydrogen ion and sodium ion concentrations and the sodium ion flux. Changes in the hydrogen ion flux were smaller and, in some experiments, insignificant. Change in the hydrogen ion flux is less sensitive than the other parameters as an indicator of minimal mucosal damage. | Some minimal effects of bile acid on canine gastric mucosa in Heidenhain pouches. The gastric mucosa in four canine Heidenhain pouches was damaged with sodium taurocholate in a concentration which was just sufficient in each dog to produce consistent changes. Solutions were placed in the pouches and changes in the volume, concentrations, and fluxes of both hydrogen ion and sodium ion were measured before and during exposure to taurocholate. Changes which occurred with taurocholate were compared with control periods; taurocholate caused a relative increase in volume of 1 ml and approximately a fourfold increase in the hydrogen ion and sodium ion concentrations and the sodium ion flux. Changes in the hydrogen ion flux were smaller and, in some experiments, insignificant. Change in the hydrogen ion flux is less sensitive than the other parameters as an indicator of minimal mucosal damage. | [
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PMID:7955 | Stability of frozen solutions of cefazolin sodium. | The stability of frozen solutions of cefazolin sodium was investigated in nine commonly used diluents at concentrations of 1 g with 2.5 ml, 500 mg with 100 ml and 10 g with 45 ml in both glass and polyvinylchloride plastic containers. The diluents were: Water for Injection USP; 0.9% Sodium Chloride Injection USP; 5% Dextrose Injection USP (D5W); D5W with 0.02% sodium bicarbonate; D5W in Lactated Ringer's Injection USP; Lactated Ringer's Injection USP; Ionosol B in D5W; Normasol M in D5W; and Plasmalyte in D5W. Frozen cefazolin sodium solutions, containing Water for Injection USP, 5% Dextrose Inection USP or 0.9% Sodium Chloride Injection USP as the diluents, retained more than 90% of labeled potency for up to 26 weeks when frozen within one hour after reconstitution and held at -10 C or -20 C. Frozen cefazolin sodium solutions, made with other diluents, were stable for up to four weeks when frozen within one hour after reconstitution and held at -10 C. | Stability of frozen solutions of cefazolin sodium. The stability of frozen solutions of cefazolin sodium was investigated in nine commonly used diluents at concentrations of 1 g with 2.5 ml, 500 mg with 100 ml and 10 g with 45 ml in both glass and polyvinylchloride plastic containers. The diluents were: Water for Injection USP; 0.9% Sodium Chloride Injection USP; 5% Dextrose Injection USP (D5W); D5W with 0.02% sodium bicarbonate; D5W in Lactated Ringer's Injection USP; Lactated Ringer's Injection USP; Ionosol B in D5W; Normasol M in D5W; and Plasmalyte in D5W. Frozen cefazolin sodium solutions, containing Water for Injection USP, 5% Dextrose Inection USP or 0.9% Sodium Chloride Injection USP as the diluents, retained more than 90% of labeled potency for up to 26 weeks when frozen within one hour after reconstitution and held at -10 C or -20 C. Frozen cefazolin sodium solutions, made with other diluents, were stable for up to four weeks when frozen within one hour after reconstitution and held at -10 C. | [
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PMID:7957 | Takayasu's arteritis and congenital coarctation of the descending thoracic and abdominal aorta: a critical review. | A critical reappraisal of the clinical, arteriographic, and pathologic features of Takayasu's arteritis and so-called congenital aortic coarctations at atypical sites is presented. It is concluded that as an isolated cardiovascular abnormality, cases of atypical congenital coarctations of the descending thoracic and abdominal aorta are probably rare. The majority of atypical aortic coarctations previously reported in the United States and Europe as congenital lesions apparently represent unrecognized cases of Takayasu's arteritis. | Takayasu's arteritis and congenital coarctation of the descending thoracic and abdominal aorta: a critical review. A critical reappraisal of the clinical, arteriographic, and pathologic features of Takayasu's arteritis and so-called congenital aortic coarctations at atypical sites is presented. It is concluded that as an isolated cardiovascular abnormality, cases of atypical congenital coarctations of the descending thoracic and abdominal aorta are probably rare. The majority of atypical aortic coarctations previously reported in the United States and Europe as congenital lesions apparently represent unrecognized cases of Takayasu's arteritis. | [
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PMID:7958 | Multiple gastric polyps and parathyroid adenomas. Report of two cases. | Two cases of multiple gastric polyps associated with parathyroid adenomas are presented. A review of the literature revealed four patients with multiple gastric polyps and multiple endocrine adenomatosis. The possiblity of multiple gastric polyps as a variant of the MEA syndrome complex is explored. Emphasis is placed on the need for thorough endocrine evaluation in patients with multiple gastric polyps. | Multiple gastric polyps and parathyroid adenomas. Report of two cases. Two cases of multiple gastric polyps associated with parathyroid adenomas are presented. A review of the literature revealed four patients with multiple gastric polyps and multiple endocrine adenomatosis. The possiblity of multiple gastric polyps as a variant of the MEA syndrome complex is explored. Emphasis is placed on the need for thorough endocrine evaluation in patients with multiple gastric polyps. | [
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PMID:7959 | Percutaneous method for arterial and venous monitoring. | A new technic for monitoring blood gases or venous and arterial pressure is described. Catheters are inserted by percutaneous puncture and guided into the femoral and iliac vessels with minimal manipulation. This rapid, simple technic has been used in more than 100 patients without a major complication. | Percutaneous method for arterial and venous monitoring. A new technic for monitoring blood gases or venous and arterial pressure is described. Catheters are inserted by percutaneous puncture and guided into the femoral and iliac vessels with minimal manipulation. This rapid, simple technic has been used in more than 100 patients without a major complication. | [
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PMID:7960 | Quantitation of local acidosis and hypoxia produced by infection. | Increased strength of healing incisions infected with E coli was demonstrated in this experiment. Efforts to measure respiratory gas tensions and pH in these incisions were unsuccessful. Therefore, these moieties were measured in normal and infected wound fluid contained in implanted wire cylinders. The wound fluid from infected cylinders was consistently more acidotic and had a lower pO2 and a higher pCO2 than fluid from unifected wound cylinders. | Quantitation of local acidosis and hypoxia produced by infection. Increased strength of healing incisions infected with E coli was demonstrated in this experiment. Efforts to measure respiratory gas tensions and pH in these incisions were unsuccessful. Therefore, these moieties were measured in normal and infected wound fluid contained in implanted wire cylinders. The wound fluid from infected cylinders was consistently more acidotic and had a lower pO2 and a higher pCO2 than fluid from unifected wound cylinders. | [
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PMID:7963 | [Controlled hypotension with sodium nitroprusside (author's transl)]. | Sodium notroprusside (Nipruss), in comparison with halothane and trimetaphan a very suitable hypotensive drug, was examined for blood pressure lowering efficency and possible side effects. For this purpose 7 patients (group 1) underwent "classical" hypotension during neurosurgical procedures. In 5 cases (group 2) nitroprusside infusion was needed for therapy of hypertensive crisis during vascular surgery. In all patients the hypotensive action was prompt and sufficient. Low blood pressure was accompanied by a 22.5% rise of heart rate and a 66% fall of central venous pressure. The development of mild metabolic acidosis seemed to be without clinical importance. The doses employed ranged from 3 to 13 (grpup 1) and from 1.5 to 3.1 gamma/kg/min (group 2). Sodium nitroprusside proved a very useful hypotensive drug, fitting well with the needs of modern anaesthesia. | [Controlled hypotension with sodium nitroprusside (author's transl)]. Sodium notroprusside (Nipruss), in comparison with halothane and trimetaphan a very suitable hypotensive drug, was examined for blood pressure lowering efficency and possible side effects. For this purpose 7 patients (group 1) underwent "classical" hypotension during neurosurgical procedures. In 5 cases (group 2) nitroprusside infusion was needed for therapy of hypertensive crisis during vascular surgery. In all patients the hypotensive action was prompt and sufficient. Low blood pressure was accompanied by a 22.5% rise of heart rate and a 66% fall of central venous pressure. The development of mild metabolic acidosis seemed to be without clinical importance. The doses employed ranged from 3 to 13 (grpup 1) and from 1.5 to 3.1 gamma/kg/min (group 2). Sodium nitroprusside proved a very useful hypotensive drug, fitting well with the needs of modern anaesthesia. | [
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PMID:7964 | [Ariway resistance in intensive care patients (author's transl)]. | With the modified interrupition technique airway resistance and tidal volume of bed-ridden patients can be measured simultaneously. This objective measure causes no distress to the patient. The effect of Noleptan on tidal volume and airway resistance has been studied in 28 patients (most of them have been unconscious). Tidal volume was improved significantly leading to an increases of arterial pO2. The bronchial airway resistance was uneffected. Our findings underline the postive experiences with Noleptan in clinical trial of intensive care. | [Ariway resistance in intensive care patients (author's transl)]. With the modified interrupition technique airway resistance and tidal volume of bed-ridden patients can be measured simultaneously. This objective measure causes no distress to the patient. The effect of Noleptan on tidal volume and airway resistance has been studied in 28 patients (most of them have been unconscious). Tidal volume was improved significantly leading to an increases of arterial pO2. The bronchial airway resistance was uneffected. Our findings underline the postive experiences with Noleptan in clinical trial of intensive care. | [
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PMID:7965 | [Studies on the stability of local anaesthetics containing adrenaline (author's transl)]. | Local toxic reactions from adrenaline following nerve block anaesthesia have been observed by many workers. Because of this potential danger we studied the changes of anaesthetic solutions containing adrenaline during a longer storage period. The measurements show a drop in the pH of the solutions caused by decomposition of adrenaline. This effect of storage is not detectable with the naked eye but makes analytical procedures necessary. Molecular oxygen and light decompose the solutions rapidly. After a storage period of some months the acidity of the solutions is comparable to concentrated acetic acid, a fact which could lead to local complications. | [Studies on the stability of local anaesthetics containing adrenaline (author's transl)]. Local toxic reactions from adrenaline following nerve block anaesthesia have been observed by many workers. Because of this potential danger we studied the changes of anaesthetic solutions containing adrenaline during a longer storage period. The measurements show a drop in the pH of the solutions caused by decomposition of adrenaline. This effect of storage is not detectable with the naked eye but makes analytical procedures necessary. Molecular oxygen and light decompose the solutions rapidly. After a storage period of some months the acidity of the solutions is comparable to concentrated acetic acid, a fact which could lead to local complications. | [
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PMID:7966 | [The calculation of the oxygen saturation as function of pO2, pH, temperature and base deviation (author's transl)]. | The paper describes a mathematical relationship between the partial pressure of oxygen and the oxygen saturation of the blood which approximates the standard oxygen dissociation curve. This approximation is based on a consideration of the electro-chemical equilibria of the reduced and the oxygenated blood. | [The calculation of the oxygen saturation as function of pO2, pH, temperature and base deviation (author's transl)]. The paper describes a mathematical relationship between the partial pressure of oxygen and the oxygen saturation of the blood which approximates the standard oxygen dissociation curve. This approximation is based on a consideration of the electro-chemical equilibria of the reduced and the oxygenated blood. | [
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PMID:7967 | Anaesthesia with sulfentanil-analgesia in carotid and vertebral arteriography. A comparison with fentanyl. | The effects of the analgesics sulfentanil (R 30730) and fentanyl, on autonimic stability, cardiovascular stability, respiratory depression and post-operative behaviour were compared in a standardised anaesthetic technique. Twenty-nine patients undergoing neurological arteriographies, were treated with one or more doses of 0.5 mg fentanyl; 22 patients were treated with an initial dose of 0.05 mg of sulfentanil followed by repeat injections of 0.025 mg, when necessary. The analgesic potency of sulfentanil proved to be 12 times that of fentanyl while (in contrast to fentanyl), the duration of the respiratory depression after sulfentanil did not differ significantly from the duration of optimal analgesia. The effects of both drugs on blood pressure and heart rate were minimal; autonomic stability was excellent. No serious side effects were seen in any of the patients. After sulfentanil analgesia the patients were more rapidly awake and lucid, than after fentanyl-analgesia. Further investigations with this new and promising drug seem to be of great interest. | Anaesthesia with sulfentanil-analgesia in carotid and vertebral arteriography. A comparison with fentanyl. The effects of the analgesics sulfentanil (R 30730) and fentanyl, on autonimic stability, cardiovascular stability, respiratory depression and post-operative behaviour were compared in a standardised anaesthetic technique. Twenty-nine patients undergoing neurological arteriographies, were treated with one or more doses of 0.5 mg fentanyl; 22 patients were treated with an initial dose of 0.05 mg of sulfentanil followed by repeat injections of 0.025 mg, when necessary. The analgesic potency of sulfentanil proved to be 12 times that of fentanyl while (in contrast to fentanyl), the duration of the respiratory depression after sulfentanil did not differ significantly from the duration of optimal analgesia. The effects of both drugs on blood pressure and heart rate were minimal; autonomic stability was excellent. No serious side effects were seen in any of the patients. After sulfentanil analgesia the patients were more rapidly awake and lucid, than after fentanyl-analgesia. Further investigations with this new and promising drug seem to be of great interest. | [
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PMID:7978 | [Hyperbaric oxygen therapy in cerebral circulatory insufficiency]. | From a study bearing upon 26 patients suffering from a cerebral circulatory insufficiency induced by a stenosis or a thrombosis, the writers analyse the part played by Hyperbare Oxygen in the neurologic evolution. The defining of the efficacy criteria enabled them to determine whenever this part was prevalent and obvious (that's to say in 20 p. 100 of the cases). However, in the other cases it was hard to decide whether Hyperbare Oxygen played any part. Only functional lesions are liable to benefit from this therapy which seems mainly useful to cover the period of circulatory adaptation at a time when supply circulations may come into play. The difficulty to appreciate the importance of supply circulations urges on to treat this type of patients early enough in a systematic way and all the more so as they are young. | [Hyperbaric oxygen therapy in cerebral circulatory insufficiency]. From a study bearing upon 26 patients suffering from a cerebral circulatory insufficiency induced by a stenosis or a thrombosis, the writers analyse the part played by Hyperbare Oxygen in the neurologic evolution. The defining of the efficacy criteria enabled them to determine whenever this part was prevalent and obvious (that's to say in 20 p. 100 of the cases). However, in the other cases it was hard to decide whether Hyperbare Oxygen played any part. Only functional lesions are liable to benefit from this therapy which seems mainly useful to cover the period of circulatory adaptation at a time when supply circulations may come into play. The difficulty to appreciate the importance of supply circulations urges on to treat this type of patients early enough in a systematic way and all the more so as they are young. | [
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PMID:7979 | [Autotransfusion in experimental hemoperitoneum. Hematologic study]. | In order to study the hematologic repercussions of pre-operative autotransfusion, ruptures were made in experimental spleens followed by retransfusion after intraperitoneal barbotage of long duration in 8 dogs. This was performed without any heparinization. The following facts could be established:--although the blood shed was rich in red globules, it presented major alterations with a loss of clotting agents and evidence of fibrinogenic degradating elements.--A study of the blood during the transfusion and after the intervention did not reveal any intravascular coagulation but only a loss of fibrinogen and thrombocytes proportional to the amount of autotransfused blood. The limit of haemostatic tolerance with no supply of external blood was an autotransfusion ranging about 75 p. 100 of the initial blood volume. | [Autotransfusion in experimental hemoperitoneum. Hematologic study]. In order to study the hematologic repercussions of pre-operative autotransfusion, ruptures were made in experimental spleens followed by retransfusion after intraperitoneal barbotage of long duration in 8 dogs. This was performed without any heparinization. The following facts could be established:--although the blood shed was rich in red globules, it presented major alterations with a loss of clotting agents and evidence of fibrinogenic degradating elements.--A study of the blood during the transfusion and after the intervention did not reveal any intravascular coagulation but only a loss of fibrinogen and thrombocytes proportional to the amount of autotransfused blood. The limit of haemostatic tolerance with no supply of external blood was an autotransfusion ranging about 75 p. 100 of the initial blood volume. | [
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PMID:7980 | [Resistance test to streptokinase. Its value in the establishing of a thrombolytic treatment using this enzyme]. | The reckoning of the rate of serous anti-streptokinase (ASK) evidenced that 8 p. 100 of the test-sample presented a distinct resistance to Streptokinase (SK), and that this percentage could be more important under certain pathological circumstances. On account of this fact and also because SK injectable preparations may offer a variation in activity, the writers advocate a "preliminary control in vitro" by means of the resistance test to streptokinase that enables one to make sure, for each patient, of the biological efficacy of the drug and to specify the starting posology. | [Resistance test to streptokinase. Its value in the establishing of a thrombolytic treatment using this enzyme]. The reckoning of the rate of serous anti-streptokinase (ASK) evidenced that 8 p. 100 of the test-sample presented a distinct resistance to Streptokinase (SK), and that this percentage could be more important under certain pathological circumstances. On account of this fact and also because SK injectable preparations may offer a variation in activity, the writers advocate a "preliminary control in vitro" by means of the resistance test to streptokinase that enables one to make sure, for each patient, of the biological efficacy of the drug and to specify the starting posology. | [
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PMID:7975 | Physostigmine reversal of sedation in parturients. | Parturients in whom meperidine HCl, propiomazine HCl, and scopolamine were used for analgesia and amnesia in labor and delivery were studied to determine the efficacy and safety of physostigmine reversal after delivery. Of a total of 120 patients, 108 received physostigmine salicylate at the completion of episiotomy closure, awakening in an average of 7.5 minutes compared with 137.8 in 12 controls. Physostigmine appears to be a safe, rapidly effective agent for reversing the prolonged somnolence of this sedation regimen. | Physostigmine reversal of sedation in parturients. Parturients in whom meperidine HCl, propiomazine HCl, and scopolamine were used for analgesia and amnesia in labor and delivery were studied to determine the efficacy and safety of physostigmine reversal after delivery. Of a total of 120 patients, 108 received physostigmine salicylate at the completion of episiotomy closure, awakening in an average of 7.5 minutes compared with 137.8 in 12 controls. Physostigmine appears to be a safe, rapidly effective agent for reversing the prolonged somnolence of this sedation regimen. | [
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PMID:7981 | [Presentation of equipment: the monitor of cerebral function]. | From a pair of parietal electrodes, an original apparatus--the monitor of cerebral function--gives a non-stop recording of the brain electric activity for frequencies ranging from 2 to 15 Hertz. It is at the moment less expensive than an ordinary EEG apparatus, easy to carry, to use and to read. The artefacts, interferences or technical defects are either totally eliminated or immediately detected thanks to a parallel graph. It is the first apparatus that has been conceived and used as a routine instrument for cerebral monitoring. It can be used in the Intensive Care Unit as well as in the Operating Theatre:--preventively, whenever there is a risk of an alteration in the cerebral circulation (CEC - extracorporal circulation - for instance - as a diagnosis, a prognosis or a therapy when there is a cerebral lesion (whatever the origin of it can be). The use of this apparatus (that has now become part of the daily routine at the London Hospital) has quickly proved indispensable and the common EEG is no longer being resorted to except upon very few occasions. | [Presentation of equipment: the monitor of cerebral function]. From a pair of parietal electrodes, an original apparatus--the monitor of cerebral function--gives a non-stop recording of the brain electric activity for frequencies ranging from 2 to 15 Hertz. It is at the moment less expensive than an ordinary EEG apparatus, easy to carry, to use and to read. The artefacts, interferences or technical defects are either totally eliminated or immediately detected thanks to a parallel graph. It is the first apparatus that has been conceived and used as a routine instrument for cerebral monitoring. It can be used in the Intensive Care Unit as well as in the Operating Theatre:--preventively, whenever there is a risk of an alteration in the cerebral circulation (CEC - extracorporal circulation - for instance - as a diagnosis, a prognosis or a therapy when there is a cerebral lesion (whatever the origin of it can be). The use of this apparatus (that has now become part of the daily routine at the London Hospital) has quickly proved indispensable and the common EEG is no longer being resorted to except upon very few occasions. | [
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PMID:7976 | Clinical evaluation of injectable lorazepam as a premedicant: the effect on recall. | A double-blind comparison of lorazepam and placebo, lorazepam and pentobarbital, and a comparison of lorazepam IV drip versus lorazepam IV bolus as a premedicant showed this new agent to offer promise as a sedative premedicant. However, the potency of the drug demands caution, and its use for outpatient premedication is not recommended. | Clinical evaluation of injectable lorazepam as a premedicant: the effect on recall. A double-blind comparison of lorazepam and placebo, lorazepam and pentobarbital, and a comparison of lorazepam IV drip versus lorazepam IV bolus as a premedicant showed this new agent to offer promise as a sedative premedicant. However, the potency of the drug demands caution, and its use for outpatient premedication is not recommended. | [
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PMID:7982 | [Value of moderate fentanyl dosage during anesthesis in abdominal surgery. Apropos of 100 cases]. | Fentanyl was used in 100 abdominal surgical interventions, combined with droperidol or with diazepan, always with good results as far as analgesia was concerned. Tensional variations that occurred during the induction were quite small and disappeared during the filling up. In the course of the intervention, tensional variations were only met with subjects suffering from high blood pressure. The respiratory depression that went with analgesia did not constitute an obstacle but made it necessary to use artificial ventilation for the intervetion. The awakening was always quick, smooth, without any vomiting and was influenced neither by the time taken up by the intervention nor by the condition of the patient. No residual respiratory depression requiring the use of an anti-morphinic was noted. At the end of the study, fentanyl appears as a powerful analgesic, easy to use and successful in all the cases of abdominal surgery. Its effect does not last, a drawback that can be avoided by the use of an intravenous drip. | [Value of moderate fentanyl dosage during anesthesis in abdominal surgery. Apropos of 100 cases]. Fentanyl was used in 100 abdominal surgical interventions, combined with droperidol or with diazepan, always with good results as far as analgesia was concerned. Tensional variations that occurred during the induction were quite small and disappeared during the filling up. In the course of the intervention, tensional variations were only met with subjects suffering from high blood pressure. The respiratory depression that went with analgesia did not constitute an obstacle but made it necessary to use artificial ventilation for the intervetion. The awakening was always quick, smooth, without any vomiting and was influenced neither by the time taken up by the intervention nor by the condition of the patient. No residual respiratory depression requiring the use of an anti-morphinic was noted. At the end of the study, fentanyl appears as a powerful analgesic, easy to use and successful in all the cases of abdominal surgery. Its effect does not last, a drawback that can be avoided by the use of an intravenous drip. | [
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PMID:7977 | Fluroxene (2, 2, 2-trifluorethyl vinyl ether) toxicity: a chemical aspect. | Fluroxene is highly toxic to several animal species. This toxicity is enhanced by induction of raised levels of hepatic microsomal enzymes. Experiments in rats are described which seek to assess the rleative contribution to this toxicity of the individual component groups of the fluroxene molecule. Though results point to the trifluoroethyl moiety of fluroxene as that aspect of the molecule most responsible for the observed mortality, reduction of the vinyl group modifies the pattern of liver injury. That the liver necrosis, manifest following fluroxene anesthesia in the presence of microsomal induction, is alone the direct cause of the acute death of experimental animals is questioned. | Fluroxene (2, 2, 2-trifluorethyl vinyl ether) toxicity: a chemical aspect. Fluroxene is highly toxic to several animal species. This toxicity is enhanced by induction of raised levels of hepatic microsomal enzymes. Experiments in rats are described which seek to assess the rleative contribution to this toxicity of the individual component groups of the fluroxene molecule. Though results point to the trifluoroethyl moiety of fluroxene as that aspect of the molecule most responsible for the observed mortality, reduction of the vinyl group modifies the pattern of liver injury. That the liver necrosis, manifest following fluroxene anesthesia in the presence of microsomal induction, is alone the direct cause of the acute death of experimental animals is questioned. | [
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PMID:7983 | [Attempt at a mathematical definition of pachycurare dosage. Application for pancuronium and AH 8165]. | Correlations among the elements indispensable to the choice of an average dose of pachycurare are established for surgical needs. Weight and duration of the surgery jointly allow to foresee with approximately 80 p. 100 accuracy the effective dose required for a surgical curarization controlled according to clinical and electromyographical criteria. After calculation of the average consumption of pachycurare per unit of time, a group of curves is established by successive integrations of additional doses representing a family of parabolae corresponding to a second-degree equation: P = alpha + beta t + gamma t2 where P: the cumulative dose for a determined period of time, per kg of body weight and thus total dose if t: the duration of the surgery, t: the time variable, alpha, beta and gamma: significant calculated coefficients. This analytical function established for the use of Pancuronium - bromide in surgery, is also being tried for the curarizing substance: AH.8165. | [Attempt at a mathematical definition of pachycurare dosage. Application for pancuronium and AH 8165]. Correlations among the elements indispensable to the choice of an average dose of pachycurare are established for surgical needs. Weight and duration of the surgery jointly allow to foresee with approximately 80 p. 100 accuracy the effective dose required for a surgical curarization controlled according to clinical and electromyographical criteria. After calculation of the average consumption of pachycurare per unit of time, a group of curves is established by successive integrations of additional doses representing a family of parabolae corresponding to a second-degree equation: P = alpha + beta t + gamma t2 where P: the cumulative dose for a determined period of time, per kg of body weight and thus total dose if t: the duration of the surgery, t: the time variable, alpha, beta and gamma: significant calculated coefficients. This analytical function established for the use of Pancuronium - bromide in surgery, is also being tried for the curarizing substance: AH.8165. | [
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PMID:7984 | [Parenteral use of a solution-emulsion following digestive surgery]. | A post-operative use of the calorico-nitrogenous mixture: Trivé 1000 was carried out in 25 patients after digestive surgery. The prescribed dose of this lipidic emulsion was 1,36 1 for an average period of four days. A study of the results led to establish the following facts:--a 55 p. cent increase in the nitrogen retention in comparison with a control series of ten patients and from (J o) the day of the intervention to (J F) the day when the treatment was stopped. Besides, the duration of the treatment did not seem to modify this balance, at least within the limits of the period of use. On the other hand, the quantity of EB 51 was a determining factor since 2000 ml were necessary to obtain a positive nitrogen balance (+ 0.96 g).--a very small rise in the amino-acid serous rate. It appeared in 14 patients and, supplied by EB 51, 3 other amino-acids increased their proportion spontaneously (Asp.--Gluc.--Tau.).--a reduction of the rate of 5 amino-acids, one of which, however, (Arginine) was being supplied by the drip. The variations were never statistically significant but for the Phenylalanine rate increase.--a non-systematic variation in the blood proteids that decreased as far as total proteids and albumin were concerned and that increased as far as globulins alpha1 and alpha2 were concerned.--Finally, no variations in the serous graphic record of lipid levels were noted, neither in the average of figures, nor in the analyses carried out after each bottle. Besides, it must be added that the tolerance of the product was excellent both on a general level (very few cases of intolerance: 4) and on the level of the plasma turbidimetry which did not reveal and chylomicrons except in one case. Therefore Trivé 1000 proved to be an interesting nutrient combinatin quite suited to a parenteral feeding because of its high caloric power. The necessity of a calories/g/nitrogen ratio from 100 to 200 corresponding to 2000 ml of EB 51 was once more evidenced. Therefore such a dose (for lack of a protein assimilation) allows for a nitrogen retention important enough to prevent an excessive consumption of the body proteinic reserves. Such doses are perfectly tolerated both on the general and metabolic levels (mainly on the lipometabolic one). | [Parenteral use of a solution-emulsion following digestive surgery]. A post-operative use of the calorico-nitrogenous mixture: Trivé 1000 was carried out in 25 patients after digestive surgery. The prescribed dose of this lipidic emulsion was 1,36 1 for an average period of four days. A study of the results led to establish the following facts:--a 55 p. cent increase in the nitrogen retention in comparison with a control series of ten patients and from (J o) the day of the intervention to (J F) the day when the treatment was stopped. Besides, the duration of the treatment did not seem to modify this balance, at least within the limits of the period of use. On the other hand, the quantity of EB 51 was a determining factor since 2000 ml were necessary to obtain a positive nitrogen balance (+ 0.96 g).--a very small rise in the amino-acid serous rate. It appeared in 14 patients and, supplied by EB 51, 3 other amino-acids increased their proportion spontaneously (Asp.--Gluc.--Tau.).--a reduction of the rate of 5 amino-acids, one of which, however, (Arginine) was being supplied by the drip. The variations were never statistically significant but for the Phenylalanine rate increase.--a non-systematic variation in the blood proteids that decreased as far as total proteids and albumin were concerned and that increased as far as globulins alpha1 and alpha2 were concerned.--Finally, no variations in the serous graphic record of lipid levels were noted, neither in the average of figures, nor in the analyses carried out after each bottle. Besides, it must be added that the tolerance of the product was excellent both on a general level (very few cases of intolerance: 4) and on the level of the plasma turbidimetry which did not reveal and chylomicrons except in one case. Therefore Trivé 1000 proved to be an interesting nutrient combinatin quite suited to a parenteral feeding because of its high caloric power. The necessity of a calories/g/nitrogen ratio from 100 to 200 corresponding to 2000 ml of EB 51 was once more evidenced. Therefore such a dose (for lack of a protein assimilation) allows for a nitrogen retention important enough to prevent an excessive consumption of the body proteinic reserves. Such doses are perfectly tolerated both on the general and metabolic levels (mainly on the lipometabolic one). | [
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PMID:7985 | [Complete parenteral alimentation after major digestive surgery using lEB 51-Trivé 1000]. | The aim of this work is to study the tolerance and the effectiveness of a complete intravenous feeding with EB 51 - Trivemil in immediate post-operative period, in 40 patients undergoing major digestive surgery. The patients were divided into three groups:--first group: 25 patients were given daily an average of 2900 calories 1500 of which via the Trivemil, during the first four post-operative days.--Second group: 5 patients received the same daily caloric dose but because of local complications, drips were maintained until the 7th day.--Third group: 10 patients received an average of 3100 calories daily 2000 of which via the Trivemil. The tolerance was evaluated by a clinical observation (blood pressure - nausea - cutaneous manifestations - temperature) and by a biological observation (hemolysis - free hemoglobin - plasma clearing). The use and effectiveness of the nutrient was evaluated - globally - by a study of body weight according to the hydro-electrolytic balance and then - individually - by the balance of the intakes and secretions of the glucides, lipids and proteids. The tolerance was excellent in all the cases, the weight remained stable, the nitrogen retention was constant mainly in the third group where it seemed to depend on the initial total energetic intake. | [Complete parenteral alimentation after major digestive surgery using lEB 51-Trivé 1000]. The aim of this work is to study the tolerance and the effectiveness of a complete intravenous feeding with EB 51 - Trivemil in immediate post-operative period, in 40 patients undergoing major digestive surgery. The patients were divided into three groups:--first group: 25 patients were given daily an average of 2900 calories 1500 of which via the Trivemil, during the first four post-operative days.--Second group: 5 patients received the same daily caloric dose but because of local complications, drips were maintained until the 7th day.--Third group: 10 patients received an average of 3100 calories daily 2000 of which via the Trivemil. The tolerance was evaluated by a clinical observation (blood pressure - nausea - cutaneous manifestations - temperature) and by a biological observation (hemolysis - free hemoglobin - plasma clearing). The use and effectiveness of the nutrient was evaluated - globally - by a study of body weight according to the hydro-electrolytic balance and then - individually - by the balance of the intakes and secretions of the glucides, lipids and proteids. The tolerance was excellent in all the cases, the weight remained stable, the nitrogen retention was constant mainly in the third group where it seemed to depend on the initial total energetic intake. | [
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PMID:7987 | [Corneal and lens deposits due to treatment by phenothiazine type neuroleptics]. | The authors have studied the corneal and lens lesions which appeared following a prolonged treatment by phenothiazines. They examined 186 patients: 147 took phenothiazines of which 35 of them presented anterior segment alterations. It seems that all phenothiazines can be held responsible for the apparition of these lesions. At this point the authors evaluated the global dose of the various phenothiazines which were administered. The threshold at which the association of these lesions seem to appear, seems to be situated around 300 gr. The total quantity of phenothiazines which are absorbed seems to be a good measure of the risk of ocular toxicity. In the case of one patient they observed that his visual keenness was lowered due to the importance of his corneal and lens lesions. | [Corneal and lens deposits due to treatment by phenothiazine type neuroleptics]. The authors have studied the corneal and lens lesions which appeared following a prolonged treatment by phenothiazines. They examined 186 patients: 147 took phenothiazines of which 35 of them presented anterior segment alterations. It seems that all phenothiazines can be held responsible for the apparition of these lesions. At this point the authors evaluated the global dose of the various phenothiazines which were administered. The threshold at which the association of these lesions seem to appear, seems to be situated around 300 gr. The total quantity of phenothiazines which are absorbed seems to be a good measure of the risk of ocular toxicity. In the case of one patient they observed that his visual keenness was lowered due to the importance of his corneal and lens lesions. | [
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PMID:7988 | [The effect of pH and partial pressure of oxygen on nitrification (author's transl)]. | A mathematical model which describes the effect of pH and dissolved oxygen on growth and activity of Nitrosomonas and Nitrobacter in mixed culture is presented. A good fit is obtained between experimental data and theoretical curves computed with parameters estimated by independent methods. Experimental data together with simulation studies show that inadequate aeration induces a temporal shift of ammonium and nitrite oxidations resulting in a transient nitrite accumulation similar to that caused by an increase of temperature. | [The effect of pH and partial pressure of oxygen on nitrification (author's transl)]. A mathematical model which describes the effect of pH and dissolved oxygen on growth and activity of Nitrosomonas and Nitrobacter in mixed culture is presented. A good fit is obtained between experimental data and theoretical curves computed with parameters estimated by independent methods. Experimental data together with simulation studies show that inadequate aeration induces a temporal shift of ammonium and nitrite oxidations resulting in a transient nitrite accumulation similar to that caused by an increase of temperature. | [
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PMID:7989 | [Nitric oxide production in rice soils (author's transl)]. | Nitric oxide gas evolution from nitrite was studied in vitro in three rice soils by gas chromatography. Autoclaved soils showed an NO evolution when supplemented with nitrite. Yet, when temperature of incubation, soil pH, soil moisture content and nitrite concentration were varied in the three soils, and with addition of nitrite reductase inhibitors, it appeared in one soil that NO production was partially a biological process. Thus, NO formation was two times as high in non-sterile soil as in sterile soil, and decreased when the temperature increased. Optimal NO production occurred at about neutrality and increased with increasing soil moisture content; moreover, this NO formation increased much less than in the other two soils with increasing nitrite concentration. Finally, the first soil contained three times more denitrifying bacteria tolerating a high nitrite concentration (5 g/1) that the other soils. | [Nitric oxide production in rice soils (author's transl)]. Nitric oxide gas evolution from nitrite was studied in vitro in three rice soils by gas chromatography. Autoclaved soils showed an NO evolution when supplemented with nitrite. Yet, when temperature of incubation, soil pH, soil moisture content and nitrite concentration were varied in the three soils, and with addition of nitrite reductase inhibitors, it appeared in one soil that NO production was partially a biological process. Thus, NO formation was two times as high in non-sterile soil as in sterile soil, and decreased when the temperature increased. Optimal NO production occurred at about neutrality and increased with increasing soil moisture content; moreover, this NO formation increased much less than in the other two soils with increasing nitrite concentration. Finally, the first soil contained three times more denitrifying bacteria tolerating a high nitrite concentration (5 g/1) that the other soils. | [
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PMID:7994 | [Effect of different pH values on the medium on the synthesis of antibiotics in the joint cultivation of Actinomyces levoris with yeasts]. | Changes in the pH level of the fermentation medium used for preliminary cultivation of C. tropicalis were studied with respect to its initial aciditv or alkalinitv. When C. tropicalis was grown on the medium used for levorin fermentation with ph 5.1--10.3, the yeast changed it in 24 hours to the level of 6.2--7.9. As dependent on the initial pH values for cultivation of C. trophicalis, production of levorin on subsequent inoculation of Act. levoris changed. The antibiotic activity increased and ranged within 120--178% of the control. Synthesis of levoristatin, a non-polyenic antibiotic equally increased under such conditions and ranged within 153--163% of the control. The pH values of 9.4--10.3 of the initial fermentation medium were optimal for mixed cultivation of Act levoris and C. tropicalis and maxium production of levorin and levoristatin. | [Effect of different pH values on the medium on the synthesis of antibiotics in the joint cultivation of Actinomyces levoris with yeasts]. Changes in the pH level of the fermentation medium used for preliminary cultivation of C. tropicalis were studied with respect to its initial aciditv or alkalinitv. When C. tropicalis was grown on the medium used for levorin fermentation with ph 5.1--10.3, the yeast changed it in 24 hours to the level of 6.2--7.9. As dependent on the initial pH values for cultivation of C. trophicalis, production of levorin on subsequent inoculation of Act. levoris changed. The antibiotic activity increased and ranged within 120--178% of the control. Synthesis of levoristatin, a non-polyenic antibiotic equally increased under such conditions and ranged within 153--163% of the control. The pH values of 9.4--10.3 of the initial fermentation medium were optimal for mixed cultivation of Act levoris and C. tropicalis and maxium production of levorin and levoristatin. | [
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PMID:7995 | [Effect of the initial pH value of the medium on the growth of Streptococcus lactis and the biosynthesis of nisin]. | The effect of the initial pH value of the medium within 4.0 to 6.6 on the growth of Str. lactis and biosynthesis of nicin was studied. It was found that at the initial pH 4.0--4.5 of the medium the growth of the culture was poor, i.e. 11--14% of the control (initiral pH 6.6). With an increase in the value of the initial pH at least to 5.0 the growth of Str. lactis also increased. At the initial pH 4.0 no biosynthesis of nicin was observed. Under the experimental conditions the antibiotic synthesis by tr. lactic started at the initial pH being equal to 4.5 and reached its maximum at pH 6.6. | [Effect of the initial pH value of the medium on the growth of Streptococcus lactis and the biosynthesis of nisin]. The effect of the initial pH value of the medium within 4.0 to 6.6 on the growth of Str. lactis and biosynthesis of nicin was studied. It was found that at the initial pH 4.0--4.5 of the medium the growth of the culture was poor, i.e. 11--14% of the control (initiral pH 6.6). With an increase in the value of the initial pH at least to 5.0 the growth of Str. lactis also increased. At the initial pH 4.0 no biosynthesis of nicin was observed. Under the experimental conditions the antibiotic synthesis by tr. lactic started at the initial pH being equal to 4.5 and reached its maximum at pH 6.6. | [
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PMID:7996 | [Study of E. coli penicillin amidase. The pH dependence of the equilibrium constant of the enzymatic hydrolysis of benzylpenicillin]. | The equilibrium constant for penicillin amidase-catalyzed hydrolysis of benzylpenicillin(Keg =3.00 +/- 0.24 x 10(-3) M at pH 5.0) and the ionization constants for phenylacetic acid (PAA) and the amino groups of 6-aminopenicillanic acid (6-APA) were determined (4.20 and 4.60 under conditions of the kinetic experiments respectively). The experimental data at pH 6.0 satisfactorily correlated with the theoretical pH-dependence for Keg constructed according to the hypothesis that benzylpenicillin synthesis has a thermodynamic optimum at pH 4.4 equal to a half-sum of the pK values for the carboxylic and amino groups of the PAA and 6-APA respectively. | [Study of E. coli penicillin amidase. The pH dependence of the equilibrium constant of the enzymatic hydrolysis of benzylpenicillin]. The equilibrium constant for penicillin amidase-catalyzed hydrolysis of benzylpenicillin(Keg =3.00 +/- 0.24 x 10(-3) M at pH 5.0) and the ionization constants for phenylacetic acid (PAA) and the amino groups of 6-aminopenicillanic acid (6-APA) were determined (4.20 and 4.60 under conditions of the kinetic experiments respectively). The experimental data at pH 6.0 satisfactorily correlated with the theoretical pH-dependence for Keg constructed according to the hypothesis that benzylpenicillin synthesis has a thermodynamic optimum at pH 4.4 equal to a half-sum of the pK values for the carboxylic and amino groups of the PAA and 6-APA respectively. | [
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PMID:7997 | Formation of beta-lactamase in Bacteroides fragilis: cell-bound and extracellular activity. | Nine strains of Bacteroides fragilis were cultivated in stirred fermentors and tested for their ability to produce beta-lactamase. There was a correlation between formation of beta-lactamase and high values of the minimal inhibitory concentration against beta-lactam antibiotics. B. fragilis strain B70 was used for optimizing the production of beta-lactamase. The highest bacterial yield was obtained in a proteose peptone-yeast extract medium. Optimal conditions for growth and beta-lactamase production were obtained at 37 C and pH 7.0. The beta-lactamase was released into the surrounding medium during the growth period to about 50%. Osmotic shock released about 20% of the total activity, and remaining activity was found in the cytoplasmic fraction. Substrate profile studies on four beta-lactamase-producing strains showed that the enzymes were mainly cephalosporinases. They are inhibited by cloxacillin, p-chloromercuribenzoate, and iodine. Analytical isoelectric focusing in polyacrylamide gel gave an isoelectric point of 4.9 +/- 0.2 for three of the strains and 5.6 +/- 0.2 for one. Comparison with beta-lactamases from aerobic gram-negative species with regard to isoelectric points showed no similarities. Also the molecular weight of the beta-lactamase from strain B70 of 43,000 indicates that this is a new class of beta-lactamase. | Formation of beta-lactamase in Bacteroides fragilis: cell-bound and extracellular activity. Nine strains of Bacteroides fragilis were cultivated in stirred fermentors and tested for their ability to produce beta-lactamase. There was a correlation between formation of beta-lactamase and high values of the minimal inhibitory concentration against beta-lactam antibiotics. B. fragilis strain B70 was used for optimizing the production of beta-lactamase. The highest bacterial yield was obtained in a proteose peptone-yeast extract medium. Optimal conditions for growth and beta-lactamase production were obtained at 37 C and pH 7.0. The beta-lactamase was released into the surrounding medium during the growth period to about 50%. Osmotic shock released about 20% of the total activity, and remaining activity was found in the cytoplasmic fraction. Substrate profile studies on four beta-lactamase-producing strains showed that the enzymes were mainly cephalosporinases. They are inhibited by cloxacillin, p-chloromercuribenzoate, and iodine. Analytical isoelectric focusing in polyacrylamide gel gave an isoelectric point of 4.9 +/- 0.2 for three of the strains and 5.6 +/- 0.2 for one. Comparison with beta-lactamases from aerobic gram-negative species with regard to isoelectric points showed no similarities. Also the molecular weight of the beta-lactamase from strain B70 of 43,000 indicates that this is a new class of beta-lactamase. | [
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PMID:7998 | Enzymatic hydrolysis of cephalosporin C by an extracellular acetylhydrolase of Cephalosporium acremonium. | Extracellular hydrolases from Cephalosporium acremonium were analyzed according to their ability to deacetylate the beta-lactam antibiotic cephalosporin C. One out of at least six hydrolases exhibits appreciable cephalosporin C acetylhydrolase (CAH) activity. This enzyme was separated from other hydrolases and purified 220-fold. The purified CAH has a relatively low affinity for cephalosporin C (K(m), 20 mM) and is strongly inhibited by diisopropylfluorophosphate and less markedly affected by fluoride. Addition of glucose, maltose, and sucrose to the culture broth suppresses CAH production, whereas glycerol and succinate have no effect. Verrucarin A prevented the enzyme from appearing in the medium, which indicates the necessity of protein synthesis for CAH formation. When 1-thio-d-glucose was added to the culture medium, the results suggested that this glucose analogue is able to inhibit CAH synthesis. Our data provide evidence for a regulation of CAH synthesis similar to the catabolite repression system in bacteria. | Enzymatic hydrolysis of cephalosporin C by an extracellular acetylhydrolase of Cephalosporium acremonium. Extracellular hydrolases from Cephalosporium acremonium were analyzed according to their ability to deacetylate the beta-lactam antibiotic cephalosporin C. One out of at least six hydrolases exhibits appreciable cephalosporin C acetylhydrolase (CAH) activity. This enzyme was separated from other hydrolases and purified 220-fold. The purified CAH has a relatively low affinity for cephalosporin C (K(m), 20 mM) and is strongly inhibited by diisopropylfluorophosphate and less markedly affected by fluoride. Addition of glucose, maltose, and sucrose to the culture broth suppresses CAH production, whereas glycerol and succinate have no effect. Verrucarin A prevented the enzyme from appearing in the medium, which indicates the necessity of protein synthesis for CAH formation. When 1-thio-d-glucose was added to the culture medium, the results suggested that this glucose analogue is able to inhibit CAH synthesis. Our data provide evidence for a regulation of CAH synthesis similar to the catabolite repression system in bacteria. | [
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PMID:7999 | Autolysis of high-GC isolates of Pseudomonas putrefaciens. | High-GC isolates of P. putrefaciens undergo extensive autolysis after growth, resulting in a marked decrease in turbidity and the release of high-molecular-weight DNA which imparts a high viscosity to culture broths. The native DNA released is resistant to attack by the exocellular DNase activity of the culture broths. Autolysis is inhibited by a pH of 6.0 and the presence of 0.001 M Mg++ or Ca++, and is enhanced by elevated pH values and temperatures. This autolytic phenomenon in broth cultures readily distinguishes high- from low-GC isolates. The latter do not exhibit autolysis. | Autolysis of high-GC isolates of Pseudomonas putrefaciens. High-GC isolates of P. putrefaciens undergo extensive autolysis after growth, resulting in a marked decrease in turbidity and the release of high-molecular-weight DNA which imparts a high viscosity to culture broths. The native DNA released is resistant to attack by the exocellular DNase activity of the culture broths. Autolysis is inhibited by a pH of 6.0 and the presence of 0.001 M Mg++ or Ca++, and is enhanced by elevated pH values and temperatures. This autolytic phenomenon in broth cultures readily distinguishes high- from low-GC isolates. The latter do not exhibit autolysis. | [
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PMID:8000 | Microbial metabolism of ethylene. | The ethylene-oxidizing strain E20 was grown on different carbon sources to obtain information on the metabolism of ethylene from simultaneous adaptation studies and from measurements of specific activities of enzymes in cell-free extracts. From the simultaneous adaptation studies it was concluded that ethylene oxide is a product of ethylene catabolism. The bacterium was also able to grow on the epoxide. From a comparison of the specific activities of isocitrate lyase and malate synthetase in different extracts it was concluded that the glyoxylate cycle was involved in the metabolism of ethylene, indicating that acetyl-CoA is a metabolite of ethylene catabolism. The sequence of reactions leading from ethylene oxide to acetyl-CoA could not be established from the simultaneous adaptation experiments and the enzyme activities in extracts. | Microbial metabolism of ethylene. The ethylene-oxidizing strain E20 was grown on different carbon sources to obtain information on the metabolism of ethylene from simultaneous adaptation studies and from measurements of specific activities of enzymes in cell-free extracts. From the simultaneous adaptation studies it was concluded that ethylene oxide is a product of ethylene catabolism. The bacterium was also able to grow on the epoxide. From a comparison of the specific activities of isocitrate lyase and malate synthetase in different extracts it was concluded that the glyoxylate cycle was involved in the metabolism of ethylene, indicating that acetyl-CoA is a metabolite of ethylene catabolism. The sequence of reactions leading from ethylene oxide to acetyl-CoA could not be established from the simultaneous adaptation experiments and the enzyme activities in extracts. | [
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PMID:8001 | Characterization of the neoagarotetra-ase and neoagarobiase of Cytophaga flevensis. | The degradation of neoagarotetraose and neoagarobiose by Cytophaga flevensis was investigated. The organism possesses an enzyme that hydrolyzes the tetramer by cleavage of its central beta-galactosidic linkage. The product of this reaction, neoagarobiose, is further hydrolyzed enzymatically to D-galactose and 3,6-anhydro-L-galactose. Both enzyme activities were localized in the cytoplasm. Attempts were made to partially purify the respective enzymes and although a 30-40 fold-purification was achieved, the final preparation contained both neoagarotetra-ase and neoagarobiase activities. Evidence was obtained that these activities were due to different enzymes. Neoagarotetra-ase is highly specific for oligosaccharides containing neoagarobiose units; the rate of hydrolysis is greatest with neoagarotetraose. It cannot hydrolyze pyruvated neoagarotetraose. Optimal conditions for its activity were pH 7.0 and 25 C. Neoagarobiase hydrolyzes only neoagarobiose and neoagarobiitol and optimal conditions for activity were pH 6.75 and 25 C. Both enzymes were inhibited by Ag+, Hg2+ and Zn2+ ions and by p-CMB, which indicates that thiol groups are present in their active centres. Both enzymes were induced by neoagaro-oligosaccharides and melibiose and were repressed when glucose was added to the medium. Neoagarobiase was also induced by D-galacturonic acid. In continuous culture, the rate of enzyme production was maximal at a dilution rate of 0.1 h-1. | Characterization of the neoagarotetra-ase and neoagarobiase of Cytophaga flevensis. The degradation of neoagarotetraose and neoagarobiose by Cytophaga flevensis was investigated. The organism possesses an enzyme that hydrolyzes the tetramer by cleavage of its central beta-galactosidic linkage. The product of this reaction, neoagarobiose, is further hydrolyzed enzymatically to D-galactose and 3,6-anhydro-L-galactose. Both enzyme activities were localized in the cytoplasm. Attempts were made to partially purify the respective enzymes and although a 30-40 fold-purification was achieved, the final preparation contained both neoagarotetra-ase and neoagarobiase activities. Evidence was obtained that these activities were due to different enzymes. Neoagarotetra-ase is highly specific for oligosaccharides containing neoagarobiose units; the rate of hydrolysis is greatest with neoagarotetraose. It cannot hydrolyze pyruvated neoagarotetraose. Optimal conditions for its activity were pH 7.0 and 25 C. Neoagarobiase hydrolyzes only neoagarobiose and neoagarobiitol and optimal conditions for activity were pH 6.75 and 25 C. Both enzymes were inhibited by Ag+, Hg2+ and Zn2+ ions and by p-CMB, which indicates that thiol groups are present in their active centres. Both enzymes were induced by neoagaro-oligosaccharides and melibiose and were repressed when glucose was added to the medium. Neoagarobiase was also induced by D-galacturonic acid. In continuous culture, the rate of enzyme production was maximal at a dilution rate of 0.1 h-1. | [
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PMID:8002 | Purification and some properties of an extracellular maltase from Bacillus subtilis. | Bacillus subtilis P-11, capable of producing extracellular maltase, was isolated from soil. Maximum enzyme production was obtained on a medium containing 2.0% methyl-alpha-D-glucose, 0.5% phytone, and 0.2% yeast extract. After the removal of cells, extracellular maltase was precipitated by ammonium sulfate (85% saturation). The enzyme was purified by using the following procedures: Sephadex G-200 column chromatography, diethylaminoethyl-Sephadex A-50 ion-exchange column chromatography, and a second Sephadex G-200 column chromatography. A highly purified maltase without amylase or proteinase activities was obtained. Some properties of the extracellular maltase were determined: optimum pH, 6.0; optimum temperature, 45 C, when the incubation time was 30 min; pH stability, within 5.5 to 6.5; heat stability, stable up to 45 C; isoelectric point, pH 6.0 (by gel-isoelectric focusing); molecular weight, 33,000 (by gel filtration with Sephadex G-200); substrate specificity: the relative rates of hydrolysis of maltose, maltotriose, isomaltose, and maltotetraose were 100:15:14:4, respectively, and there was no activity toward alkyl or aryl-alpha-D-glucosides, amylose, or other higher polymers. Transglucosylase activity was present. Glucose and tris(hydroxymethyl)aminomethane were competitive inhibitors with Ki values of 4.54 and 75.08 mM, respectively; cysteine was a noncompetitive inhibitor. Michaelis constants were 5 mM for maltose, 1 mM for maltoriose, and 10 mM for isomaltose. A plot of pKm (-log Km) versus pH revealed two deflection points, one each at 5.5 and 6.5; these probably corresponded to an imidazole group of a histidine residue in or near the active center; this assumption was supported by the strong inhibition of enzyme activity by rose bengal. | Purification and some properties of an extracellular maltase from Bacillus subtilis. Bacillus subtilis P-11, capable of producing extracellular maltase, was isolated from soil. Maximum enzyme production was obtained on a medium containing 2.0% methyl-alpha-D-glucose, 0.5% phytone, and 0.2% yeast extract. After the removal of cells, extracellular maltase was precipitated by ammonium sulfate (85% saturation). The enzyme was purified by using the following procedures: Sephadex G-200 column chromatography, diethylaminoethyl-Sephadex A-50 ion-exchange column chromatography, and a second Sephadex G-200 column chromatography. A highly purified maltase without amylase or proteinase activities was obtained. Some properties of the extracellular maltase were determined: optimum pH, 6.0; optimum temperature, 45 C, when the incubation time was 30 min; pH stability, within 5.5 to 6.5; heat stability, stable up to 45 C; isoelectric point, pH 6.0 (by gel-isoelectric focusing); molecular weight, 33,000 (by gel filtration with Sephadex G-200); substrate specificity: the relative rates of hydrolysis of maltose, maltotriose, isomaltose, and maltotetraose were 100:15:14:4, respectively, and there was no activity toward alkyl or aryl-alpha-D-glucosides, amylose, or other higher polymers. Transglucosylase activity was present. Glucose and tris(hydroxymethyl)aminomethane were competitive inhibitors with Ki values of 4.54 and 75.08 mM, respectively; cysteine was a noncompetitive inhibitor. Michaelis constants were 5 mM for maltose, 1 mM for maltoriose, and 10 mM for isomaltose. A plot of pKm (-log Km) versus pH revealed two deflection points, one each at 5.5 and 6.5; these probably corresponded to an imidazole group of a histidine residue in or near the active center; this assumption was supported by the strong inhibition of enzyme activity by rose bengal. | [
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PMID:8003 | Influence of growth temperature on glucose metabolism of a psychotrophic strain of Bacillus cereus. | The influence of temperature on glucose metabolism of a psychotrophic strain of Bacillus cereus was investigated. The pH of the growth medium and spore-forming frequencies of B. cereus varied when grown at 32, 20, or 7 C. Radiorespirometric analyses revealed that vegetative cells of B. cereus metabolized glucose by simultaneous operation of the Embden-Meyerhof-Parnas pathway and the pentose phosphate pathway. As the growth temperature decreased, glucose was metabolized with increased participation of the pentose phosphate pathway. The shift of cells grown at a higher temperature to a lower temperature increased the relative participation of the pentose phosphate pathway, whereas the shift of cells grown at low temperatures to a higher temperature had the opposite effect. Cells of late logarithmic phase grown at 20 and 7 C oxidized acetate by the tricarboxylic acid cycle reaction. However, cells grown at 32 C failed to oxidize acetate to CO2 to any appreciable extent. The extracellular products resulting from the metabolism of glucose decreased as the growth temperature was lowered. Organic acids were the major extracellular products of cultures grown at 32 and 20 C. Acetic acid, lactic acid, and pyruvic acid together accounted for 86.1 and 78.9% of extracellular radioactivity, respectively, at the two temperatures. The relative ratio of these three acids varied between the temperatures. Little or no acid accumulated at 7 C. | Influence of growth temperature on glucose metabolism of a psychotrophic strain of Bacillus cereus. The influence of temperature on glucose metabolism of a psychotrophic strain of Bacillus cereus was investigated. The pH of the growth medium and spore-forming frequencies of B. cereus varied when grown at 32, 20, or 7 C. Radiorespirometric analyses revealed that vegetative cells of B. cereus metabolized glucose by simultaneous operation of the Embden-Meyerhof-Parnas pathway and the pentose phosphate pathway. As the growth temperature decreased, glucose was metabolized with increased participation of the pentose phosphate pathway. The shift of cells grown at a higher temperature to a lower temperature increased the relative participation of the pentose phosphate pathway, whereas the shift of cells grown at low temperatures to a higher temperature had the opposite effect. Cells of late logarithmic phase grown at 20 and 7 C oxidized acetate by the tricarboxylic acid cycle reaction. However, cells grown at 32 C failed to oxidize acetate to CO2 to any appreciable extent. The extracellular products resulting from the metabolism of glucose decreased as the growth temperature was lowered. Organic acids were the major extracellular products of cultures grown at 32 and 20 C. Acetic acid, lactic acid, and pyruvic acid together accounted for 86.1 and 78.9% of extracellular radioactivity, respectively, at the two temperatures. The relative ratio of these three acids varied between the temperatures. Little or no acid accumulated at 7 C. | [
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PMID:8004 | Inhibition of Clostridium perfringens by heated combinations of nitrite, sulfur, and ferrous or ferric ions. | Heating mixtures of sodium nitrite, cysteine, and either ferrous sulfate or ferric chloride at 121 C for 20 min at pH 6.5 or 6.3 produced a potent inhibitor of Clostridium perfringens vegetative cells and spores when added to previously heat-sterilized fluid thioglycolate medium. When the mixtures containing FeSO4 at pH 5.2 or FeCl3 at pH 2.7 were heated, the inhibitory effect was not produced. These responses seem to eliminate the possibility that cysteine nitrosothiol is the agent responsible for the heated-nitrite inhibition known as the Perigo effect. The variable pH responses also cast doubt upon the role of the black Roussin salt as the agent of the Perigo effect. | Inhibition of Clostridium perfringens by heated combinations of nitrite, sulfur, and ferrous or ferric ions. Heating mixtures of sodium nitrite, cysteine, and either ferrous sulfate or ferric chloride at 121 C for 20 min at pH 6.5 or 6.3 produced a potent inhibitor of Clostridium perfringens vegetative cells and spores when added to previously heat-sterilized fluid thioglycolate medium. When the mixtures containing FeSO4 at pH 5.2 or FeCl3 at pH 2.7 were heated, the inhibitory effect was not produced. These responses seem to eliminate the possibility that cysteine nitrosothiol is the agent responsible for the heated-nitrite inhibition known as the Perigo effect. The variable pH responses also cast doubt upon the role of the black Roussin salt as the agent of the Perigo effect. | [
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PMID:8005 | Pathways of microbial metabolism of parathion. | A mixed bacterial culture, consisting of a minimum of nine isolates, was adapted to growth on technical parathion (PAR) as a sole carbon and energy source. The primary oxidative pathway for PAR metabolism involved an initial hydrolysis to yield diethylthiophosphoric acid and p-nitrophenol. A secondary oxidative pathway involved the oxidation of PAR to paraoxon and then hydrolysis to yield p-nitrophenol and diethylphosphoric acid. Under low oxgen conditions PAR was reduced via a third pathway to p-aminoparathion and subsequently hydrolyzed to p-aminophenol and diethylthiophosphoric acid. PAR hydrolase, an enzyme produced by an isolate from the mixed culture, rapidly hydrolyzed PAR and paraoxon (6.0 mumol/mg per min). This enzyme was inducible and stable at room temperature and retained 100% of its activity when heated for 55 C for 10 min. | Pathways of microbial metabolism of parathion. A mixed bacterial culture, consisting of a minimum of nine isolates, was adapted to growth on technical parathion (PAR) as a sole carbon and energy source. The primary oxidative pathway for PAR metabolism involved an initial hydrolysis to yield diethylthiophosphoric acid and p-nitrophenol. A secondary oxidative pathway involved the oxidation of PAR to paraoxon and then hydrolysis to yield p-nitrophenol and diethylphosphoric acid. Under low oxgen conditions PAR was reduced via a third pathway to p-aminoparathion and subsequently hydrolyzed to p-aminophenol and diethylthiophosphoric acid. PAR hydrolase, an enzyme produced by an isolate from the mixed culture, rapidly hydrolyzed PAR and paraoxon (6.0 mumol/mg per min). This enzyme was inducible and stable at room temperature and retained 100% of its activity when heated for 55 C for 10 min. | [
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PMID:8006 | Microbiological oxidation of synthetic chalcocite and covellite by Thiobacillus ferrooxidans. | The microbiological oxidation of synthetic chalcocite and covellite has been investigated using an adapted strain of Thiobacillus ferrooxidans. Biodegradation of chalcocite was found to be 90 to 100% and that of covellite 45 to 60%. Optimum conditions for the oxidation of chalcocite were: pH, 1.7 to 2.3; temperature, 35 C; and ferric iron concentration in the range of 0.004 to 0.01 M. For covellite, the optimum conditions were: pH 2.3; temperature, 35 C; and ferric iron concentration in the range of 0.004 to 0.02 M. The energies of activation were determined to be 16.3 kcal (ca. 6.8 X 10(4) J) per mol and 11.7 kcal (ca. 4.8 X 10(4) J) per mol for chalcocite and covellite, respectively. | Microbiological oxidation of synthetic chalcocite and covellite by Thiobacillus ferrooxidans. The microbiological oxidation of synthetic chalcocite and covellite has been investigated using an adapted strain of Thiobacillus ferrooxidans. Biodegradation of chalcocite was found to be 90 to 100% and that of covellite 45 to 60%. Optimum conditions for the oxidation of chalcocite were: pH, 1.7 to 2.3; temperature, 35 C; and ferric iron concentration in the range of 0.004 to 0.01 M. For covellite, the optimum conditions were: pH 2.3; temperature, 35 C; and ferric iron concentration in the range of 0.004 to 0.02 M. The energies of activation were determined to be 16.3 kcal (ca. 6.8 X 10(4) J) per mol and 11.7 kcal (ca. 4.8 X 10(4) J) per mol for chalcocite and covellite, respectively. | [
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PMID:8016 | Marrow transplantation in treatment of children with aplastic anaemia or acute leukaemia. | Seventy-six patients, aged 2 to 17 years, were treated with bone marrow transplantation for severe aplastic anaemia or acute leukaemia refractory to conventional therapy. 16 of the 22 patients (73%) who received marrow transplantations for aplastic anaemia are surviving, 12 of these for over one year. In acute leukaemia, using preparation with cyclophosphamide and total body irradiation, 8 of 33 patients (24%) receiving allogeneic and 5 of 8 (63%) receiving syngeneic transplantations are continuing in remission from 3 months to beyond 2 years. The longest continuing remission off therapy is now over 4 1/2 years after preparation with total body irradiation. The major causes of failure remain graft-versus-host disease, infection, graft rejection (aplastic anaemia), and leukaemic relapse. | Marrow transplantation in treatment of children with aplastic anaemia or acute leukaemia. Seventy-six patients, aged 2 to 17 years, were treated with bone marrow transplantation for severe aplastic anaemia or acute leukaemia refractory to conventional therapy. 16 of the 22 patients (73%) who received marrow transplantations for aplastic anaemia are surviving, 12 of these for over one year. In acute leukaemia, using preparation with cyclophosphamide and total body irradiation, 8 of 33 patients (24%) receiving allogeneic and 5 of 8 (63%) receiving syngeneic transplantations are continuing in remission from 3 months to beyond 2 years. The longest continuing remission off therapy is now over 4 1/2 years after preparation with total body irradiation. The major causes of failure remain graft-versus-host disease, infection, graft rejection (aplastic anaemia), and leukaemic relapse. | [
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PMID:8017 | Human skin proteases. Separation and characterization of two acid proteases resembling cathepsin B1 and cathepsin D and of an inhibitor of cathepsin B1. | Two acid proteases, one hydrolysing hemoglobin and the other hydrolysing benzoyl arginine naphthyamide (BANA), were separated and partially purified from human skin buffer extract. The acid protease hydrolysing hemoglobin was purified about 190 fold by Sephadex G-100 gel filtration and DEAE-cellulose chromatography. It hydrolysed hemoglobin at pH 3.5, casein at pH 5.8 and skin protein substrate at pH 6.0. It did not markedly hydrolyse synthetic protease substrates. The molecular size of this protease was 38000. The protease was insensitive to common protease modifiers and closely resembles cathepsin D purified from other organs. The BANA-hydrolysing acid protease was purified about 760 fold by Sephadex G-100 gel filtration and affinity chromatography on organomercurial Sepharose 4B gel. It preferentially hydrolysed BAEE, BANA and BAA with an optimum at pH 5.8. The hydrolysis of BAPA, LeuNA and protein substrates was very low. This acid protease was found to be highly dependent on reducing agents, as DTT, and chelating agents, as EDTA, and was inhibited by pCMB and TLCK. The molecular size of the enzyme was 28000. This protease closely resembles cathepsin B1 purified from other organs. Human skin was also shown to contain a low activity of benzoyl arginine amide (BAA) hydrolysing acid protease with a molecular size of about 50000 and resembling cathepsin B2. Human skin contained an inhibitor with a molecular size of about 13000 against human skin cathepsin B1. This inhibitor did not inhibit trypsin, chymotrypsin or skin proteases other than cathepsin B1. | Human skin proteases. Separation and characterization of two acid proteases resembling cathepsin B1 and cathepsin D and of an inhibitor of cathepsin B1. Two acid proteases, one hydrolysing hemoglobin and the other hydrolysing benzoyl arginine naphthyamide (BANA), were separated and partially purified from human skin buffer extract. The acid protease hydrolysing hemoglobin was purified about 190 fold by Sephadex G-100 gel filtration and DEAE-cellulose chromatography. It hydrolysed hemoglobin at pH 3.5, casein at pH 5.8 and skin protein substrate at pH 6.0. It did not markedly hydrolyse synthetic protease substrates. The molecular size of this protease was 38000. The protease was insensitive to common protease modifiers and closely resembles cathepsin D purified from other organs. The BANA-hydrolysing acid protease was purified about 760 fold by Sephadex G-100 gel filtration and affinity chromatography on organomercurial Sepharose 4B gel. It preferentially hydrolysed BAEE, BANA and BAA with an optimum at pH 5.8. The hydrolysis of BAPA, LeuNA and protein substrates was very low. This acid protease was found to be highly dependent on reducing agents, as DTT, and chelating agents, as EDTA, and was inhibited by pCMB and TLCK. The molecular size of the enzyme was 28000. This protease closely resembles cathepsin B1 purified from other organs. Human skin was also shown to contain a low activity of benzoyl arginine amide (BAA) hydrolysing acid protease with a molecular size of about 50000 and resembling cathepsin B2. Human skin contained an inhibitor with a molecular size of about 13000 against human skin cathepsin B1. This inhibitor did not inhibit trypsin, chymotrypsin or skin proteases other than cathepsin B1. | [
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PMID:8021 | Hypotensive action of propranolol and a new beta-blocking agent, D-32 in conscious normotensive and renal hypertensive dogs. | The hypotensive actions of dl-propranolol and a new beta-blocking agent, dl-tert-butylamino-3-(2', 3'-dimethylphenoxy)-2-propanol hydrochloride (D-32) were studied in conscious normotensive and renal hypertensive dogs, using a cross-over design. The effects were compared with that of placebo (lactose 200 mg/head, p.o.) administered in a blind fashion. A marked reduction in systolic blood pressure, from 158 +/- 2.9 to 124 +/- 2.3 (mean +/- S.E.) mm Hg, was observed at 3 hr after administration of 50 mg/kg, p.o. of D-32 in renal hypertensive dogs but not in normotensive ones. In both normotensive and renal hypertensive dogs, 10 and 50 mg/kg, p.o. of D-32 caused a marked increase in heart rate. Intravenous infusion of p-OH D-32, a main metabolite of D-32 in dogs, at a rate of 1 mg/kg per min for 5 min into renal hypertensive dogs caused a significant, long-lasting fall in blood pressure and a sustained increase in heart rate, whereas that of D-32 (1 mg/kg per min for 5 min) failed to do so. Propranolol (10 and/or 50 mg/kg, p.o.) produced no significant changes in blood pressure and heart rate in both preparations. These results indicate that D-32 causes a fall in blood pressure in conscious renal hypertensive dogs mainly by a metabolite of D-32, p-OH D-32. | Hypotensive action of propranolol and a new beta-blocking agent, D-32 in conscious normotensive and renal hypertensive dogs. The hypotensive actions of dl-propranolol and a new beta-blocking agent, dl-tert-butylamino-3-(2', 3'-dimethylphenoxy)-2-propanol hydrochloride (D-32) were studied in conscious normotensive and renal hypertensive dogs, using a cross-over design. The effects were compared with that of placebo (lactose 200 mg/head, p.o.) administered in a blind fashion. A marked reduction in systolic blood pressure, from 158 +/- 2.9 to 124 +/- 2.3 (mean +/- S.E.) mm Hg, was observed at 3 hr after administration of 50 mg/kg, p.o. of D-32 in renal hypertensive dogs but not in normotensive ones. In both normotensive and renal hypertensive dogs, 10 and 50 mg/kg, p.o. of D-32 caused a marked increase in heart rate. Intravenous infusion of p-OH D-32, a main metabolite of D-32 in dogs, at a rate of 1 mg/kg per min for 5 min into renal hypertensive dogs caused a significant, long-lasting fall in blood pressure and a sustained increase in heart rate, whereas that of D-32 (1 mg/kg per min for 5 min) failed to do so. Propranolol (10 and/or 50 mg/kg, p.o.) produced no significant changes in blood pressure and heart rate in both preparations. These results indicate that D-32 causes a fall in blood pressure in conscious renal hypertensive dogs mainly by a metabolite of D-32, p-OH D-32. | [
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PMID:8022 | Inhibition by suprofen and other non-narcotic analgesic drugs of the effects of prostaglandin precursor on isolated tissues and platelets. | Contractions caused by Slow Reacting Substance C(SRS-C) and by Arachidonic Acid hydroperoxide (AAP) in the guinea-pig ileum and by AAP in the rat fundus were studied in the presence of suprofen and of 3 reference compounds. The dose-related inhibitions were not due to antagonism of prostaglandins. Other agonists of gastrointestinal smooth muscle were not or only weakly antagonized. For the study of selective inhibition of AAP-induced contractions by non-narcotic analgesics, the rat fundus is the preferred preparation. In this model suprofen had an ED50 of 1.27 x 10(-7) M (0.033 mug/ml), being 1.5, 94 and 2, 020 times more potent than indomethacin, phenybutazone and acetylsalicylic acid, respectively. Suprofen also strongly inhibited malondialdehyde formation by guinea-pig platelets incubated with arachidonic acid. The reported effects point to inhibition by suprofen of prostaglandin biosynthesis. The antagonism of AAP-induced contractions in the rat fundus is a valuable test system for inhibitors of prostaglandin biosynthesis. | Inhibition by suprofen and other non-narcotic analgesic drugs of the effects of prostaglandin precursor on isolated tissues and platelets. Contractions caused by Slow Reacting Substance C(SRS-C) and by Arachidonic Acid hydroperoxide (AAP) in the guinea-pig ileum and by AAP in the rat fundus were studied in the presence of suprofen and of 3 reference compounds. The dose-related inhibitions were not due to antagonism of prostaglandins. Other agonists of gastrointestinal smooth muscle were not or only weakly antagonized. For the study of selective inhibition of AAP-induced contractions by non-narcotic analgesics, the rat fundus is the preferred preparation. In this model suprofen had an ED50 of 1.27 x 10(-7) M (0.033 mug/ml), being 1.5, 94 and 2, 020 times more potent than indomethacin, phenybutazone and acetylsalicylic acid, respectively. Suprofen also strongly inhibited malondialdehyde formation by guinea-pig platelets incubated with arachidonic acid. The reported effects point to inhibition by suprofen of prostaglandin biosynthesis. The antagonism of AAP-induced contractions in the rat fundus is a valuable test system for inhibitors of prostaglandin biosynthesis. | [
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PMID:8018 | Some natural products from two soft coals. Their removal, metal-binding and enzyme inhibitory activity. | The chemical constituents of coal have not been fully characterized in relation to the incidence of coal workers' pneumoconiosis (CWP). In this study two soft coals obtained from mines in which workers had high and low incidences of CWP were leached with aqueous base and acid to remove their acidic and basic components. The results suggest that humic substances similar to those found in soil are present in the coal samples. Further, differences in the quantity of material removed ant its metal-binding and enzyme inhibitory activity are related to disease incidence. | Some natural products from two soft coals. Their removal, metal-binding and enzyme inhibitory activity. The chemical constituents of coal have not been fully characterized in relation to the incidence of coal workers' pneumoconiosis (CWP). In this study two soft coals obtained from mines in which workers had high and low incidences of CWP were leached with aqueous base and acid to remove their acidic and basic components. The results suggest that humic substances similar to those found in soil are present in the coal samples. Further, differences in the quantity of material removed ant its metal-binding and enzyme inhibitory activity are related to disease incidence. | [
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PMID:8023 | Discriminative stimulus properties of analgesic drugs: narcotic versus non-narcotic analgesics. | Using a food-reinforced two-lever operant procedure, rats (n=6) were trained to discriminate fentanyl (1.25 mg/kg, p.o., t-60') from solvent (1 ml/100 g B.W., p.o., t-60'). The administration of another narcotic analgesic (pethidine) produced a dose-related generalization with the standard fentanyl treatment; six non-narcotic analygesics (suprofen, acetylsalicylic acid, indomethacin, phenacetin, phenylbutazone, tolmetin) were found not to do so. It is concluded that the ability of drugs to produce analgesia is not a sufficient condition for the drugs to produce the narcotic cue as well. | Discriminative stimulus properties of analgesic drugs: narcotic versus non-narcotic analgesics. Using a food-reinforced two-lever operant procedure, rats (n=6) were trained to discriminate fentanyl (1.25 mg/kg, p.o., t-60') from solvent (1 ml/100 g B.W., p.o., t-60'). The administration of another narcotic analgesic (pethidine) produced a dose-related generalization with the standard fentanyl treatment; six non-narcotic analygesics (suprofen, acetylsalicylic acid, indomethacin, phenacetin, phenylbutazone, tolmetin) were found not to do so. It is concluded that the ability of drugs to produce analgesia is not a sufficient condition for the drugs to produce the narcotic cue as well. | [
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PMID:8024 | Comparative doses and costs of antipsychotic medication. | It is clinically useful to have a table listing the equivalent doses of the various neuroleptics to a standard, such as chlorpromazine. This article derives such data by reviewing double-blind controlled studies that used a flexible dosage schedule of neuroleptics in treating schizophrenic patients. Each neuroleptic is then converted to 100-mg chlorpromazine equivalents. This empirically derived dosage comparability table is compared with a similar table derived from the opinions of experts. Since these comparable doses produce equivalent amounts of antipsychotic activity, the cost to provide such medication was then calculated, and a table comparing the costs of the different neuroleptics was constructed. In absolute amounts, the cost differences between drugs are small. However, for any drug, large savings accrue when the largest possible capsule or tablet to achieve the desired dose is prescribed. | Comparative doses and costs of antipsychotic medication. It is clinically useful to have a table listing the equivalent doses of the various neuroleptics to a standard, such as chlorpromazine. This article derives such data by reviewing double-blind controlled studies that used a flexible dosage schedule of neuroleptics in treating schizophrenic patients. Each neuroleptic is then converted to 100-mg chlorpromazine equivalents. This empirically derived dosage comparability table is compared with a similar table derived from the opinions of experts. Since these comparable doses produce equivalent amounts of antipsychotic activity, the cost to provide such medication was then calculated, and a table comparing the costs of the different neuroleptics was constructed. In absolute amounts, the cost differences between drugs are small. However, for any drug, large savings accrue when the largest possible capsule or tablet to achieve the desired dose is prescribed. | [
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PMID:8025 | Pharmacokinetics of red blood cell phenothiazine and clinical effects. Acute dystonic reactions. | Pharmacokinetics of the phenothiazine, butaperazine, were studied in relationship to acute dystonic reactions. Dystonias appeared on falling drug concentrations, more than one half-life after plasma and red blood cell (RBC) peak butaperazine concentrations. Red blood cell butaperazine kinetics differentiated better than did plasma butaperazine levels those subjects in whom dystonias would develop from those in whom they did not. We conclude that RBC phenothiazine levels may more clearly reflect drug concentration at critical brain sites than do simple plasma drug levels. Furthermore, dystonic reactions may be the result of differential sensitivity of two or more receptor systems to receptor blockade by antischizophrenic agents. | Pharmacokinetics of red blood cell phenothiazine and clinical effects. Acute dystonic reactions. Pharmacokinetics of the phenothiazine, butaperazine, were studied in relationship to acute dystonic reactions. Dystonias appeared on falling drug concentrations, more than one half-life after plasma and red blood cell (RBC) peak butaperazine concentrations. Red blood cell butaperazine kinetics differentiated better than did plasma butaperazine levels those subjects in whom dystonias would develop from those in whom they did not. We conclude that RBC phenothiazine levels may more clearly reflect drug concentration at critical brain sites than do simple plasma drug levels. Furthermore, dystonic reactions may be the result of differential sensitivity of two or more receptor systems to receptor blockade by antischizophrenic agents. | [
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PMID:8026 | Takayasu arteritis. An arteriographic-pathological correlation. | We report an aortographic-pathological correlation in a patient with Takayasu arteritis. The inflammatory activity in Takayasu arteritis gradually subsides and accordingly, the histological appearance goes through a cycle of changes, ranging from acute florid inflammation to an old scarred vessel. At gross pathological inspection, the aortic intima frequently shows longitudinal wrinkling and tree-barking indistinguishable from syphilitic aortitis. In other instances, secondary atherosclerosis totally obscures the underlying changes of aortitis. In such instances, close search at various levels of the aorta is likely to uncover persistent foci of arteritis and thereby permit identification of the pathological changes as secondary to previous aortitis. | Takayasu arteritis. An arteriographic-pathological correlation. We report an aortographic-pathological correlation in a patient with Takayasu arteritis. The inflammatory activity in Takayasu arteritis gradually subsides and accordingly, the histological appearance goes through a cycle of changes, ranging from acute florid inflammation to an old scarred vessel. At gross pathological inspection, the aortic intima frequently shows longitudinal wrinkling and tree-barking indistinguishable from syphilitic aortitis. In other instances, secondary atherosclerosis totally obscures the underlying changes of aortitis. In such instances, close search at various levels of the aorta is likely to uncover persistent foci of arteritis and thereby permit identification of the pathological changes as secondary to previous aortitis. | [
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PMID:8027 | Paragangliomatosis associated with multiple endocrine adenomas. | A 19-year-old woman had multiple functioning extra-adrenal paragangliomas, a pituitary adenoma associated with acromegaly, parathyroid hyperplasia, and pigmentary abnormalities. This case differs from previously described instances of multiple endocrine adenomatosis (MEA) and has features that bridge the classic MEA type 1 and 2 syndromes and possibly Von Recklinghausen disease. The coexistence of pheochromocytoma with acromegaly is extremely rare, and the association with extra-adrenal paragangiliomas appears to be unique. Thyroid parafollicular cell proliferation could not be proved by immunohistochemical or electron microscopical studies. The large number and extensive distribution of paragangliomas, ranging from neck to pelvis, is another unique feature of this case. The concept of neurocrestopathy or of an endocrine polypeptide (APUD) cell system may offer an explanation for the interrelation of these diverse growths. | Paragangliomatosis associated with multiple endocrine adenomas. A 19-year-old woman had multiple functioning extra-adrenal paragangliomas, a pituitary adenoma associated with acromegaly, parathyroid hyperplasia, and pigmentary abnormalities. This case differs from previously described instances of multiple endocrine adenomatosis (MEA) and has features that bridge the classic MEA type 1 and 2 syndromes and possibly Von Recklinghausen disease. The coexistence of pheochromocytoma with acromegaly is extremely rare, and the association with extra-adrenal paragangiliomas appears to be unique. Thyroid parafollicular cell proliferation could not be proved by immunohistochemical or electron microscopical studies. The large number and extensive distribution of paragangliomas, ranging from neck to pelvis, is another unique feature of this case. The concept of neurocrestopathy or of an endocrine polypeptide (APUD) cell system may offer an explanation for the interrelation of these diverse growths. | [
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PMID:8028 | A case of pneumococcal typhlitis. | A 29-year-old man had abdominal pain for 24 hours. This and the results of an abdominal examination were typical of acute appendicitis. He had suffered sinusitis for two weeks. At operation, the appendix was normal; there was an abscess in the cecal wall, the exudate of which grew pneumococci. Incidental appendectomy was done and the patient was treated successfully with lincomycin hydrochloride, and later, cephalexin monohydrate. It is possible that the typhlitis was secondary to the upper respiratory infection. | A case of pneumococcal typhlitis. A 29-year-old man had abdominal pain for 24 hours. This and the results of an abdominal examination were typical of acute appendicitis. He had suffered sinusitis for two weeks. At operation, the appendix was normal; there was an abscess in the cecal wall, the exudate of which grew pneumococci. Incidental appendectomy was done and the patient was treated successfully with lincomycin hydrochloride, and later, cephalexin monohydrate. It is possible that the typhlitis was secondary to the upper respiratory infection. | [
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PMID:8032 | Analgesic nephropathy. | Analgesic nephropathy occurs most commonly in Australia, where it is the second most frequent cause of renal failure. Whilst the reasons for widespread abuse of analgesics are poorly understood, the consequences of abuse are now well recognised. Increasing emphasis is being placed on the analgesic syndrome and on the accelerated atherosclerosis seen in these patients. Attention in the article is drawn to ways in which the analgesic syndrome can be recognised and analgesic abuse ceased. A therapeutic approach is presented which allows the successful withdrawal of analgesic in almost all patients. | Analgesic nephropathy. Analgesic nephropathy occurs most commonly in Australia, where it is the second most frequent cause of renal failure. Whilst the reasons for widespread abuse of analgesics are poorly understood, the consequences of abuse are now well recognised. Increasing emphasis is being placed on the analgesic syndrome and on the accelerated atherosclerosis seen in these patients. Attention in the article is drawn to ways in which the analgesic syndrome can be recognised and analgesic abuse ceased. A therapeutic approach is presented which allows the successful withdrawal of analgesic in almost all patients. | [
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PMID:8034 | A study of the surface pH of the urinary bladder during distension in rabbits. | Bladder wall surface pH has been continuously monitored with a glass pH electrode using a simple, atraumatic technique. Experiments have been performed in the rabbit showing that there is a progressive fall in bladder wall pH as the intravesical pressure is increased. The changes in bladder wall pH serve as a sensitive indicator of bladder blood flow and so this simple method is used to assess the changes in bladder blood flow during distension. | A study of the surface pH of the urinary bladder during distension in rabbits. Bladder wall surface pH has been continuously monitored with a glass pH electrode using a simple, atraumatic technique. Experiments have been performed in the rabbit showing that there is a progressive fall in bladder wall pH as the intravesical pressure is increased. The changes in bladder wall pH serve as a sensitive indicator of bladder blood flow and so this simple method is used to assess the changes in bladder blood flow during distension. | [
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PMID:8035 | The purification and properties of the glutamine synthetase from the cytosol of Soya-bean root nodules. | The major portion of glutamine synthetase activity in root nodules of soya-bean plants is associated with the cytosol rather than with Rhizobium japonicum bacteroids. Glutamine synthetase accounts for about 2% of the total soluble protein in nodule cytosol. Glutamine synthetase from nodule cytosol has been purified by a procedure involving fractionation with protamine sulphate, ammonium sulphate and polypropylene glycol, chromatography on DEAE-Bio-Gel A and Bio-Gel A-5m and affinity chromatography on glutamate-agarose columns. The purified preparation appeared to be homogeneous in the analytical ultracentrifuge. From sedimentation-equilibrium experiments a mol. wt. of about 376000 was determined for the native enzyme and 47300 for the enzyme in guanidinium chloride. From these data and measurements of electron micrographs, we have concluded that glutamine synthetase from nodule cytosol consists of eight subunits arranged in two sets of planar tetramers which form a cubical configuration with dimensions of about 10 nm (100 A) across each side. Glutamine synthetase from nodule cytosol has a higher glycine and proline content and a lower content of phenylalanine than the glutamine synthetase that has been prepared from pea seed. The cytosol enzyme contains four half-cystine molecules per subunit, which is in contrast with two reported for the enzyme from pea seed. Enzyme activity is striking influenced by the relative proportion of Mg2+ and Mn2+ in the assay medium. Activity is inhibited by feedback inhibitors and is influenced by energy charge. | The purification and properties of the glutamine synthetase from the cytosol of Soya-bean root nodules. The major portion of glutamine synthetase activity in root nodules of soya-bean plants is associated with the cytosol rather than with Rhizobium japonicum bacteroids. Glutamine synthetase accounts for about 2% of the total soluble protein in nodule cytosol. Glutamine synthetase from nodule cytosol has been purified by a procedure involving fractionation with protamine sulphate, ammonium sulphate and polypropylene glycol, chromatography on DEAE-Bio-Gel A and Bio-Gel A-5m and affinity chromatography on glutamate-agarose columns. The purified preparation appeared to be homogeneous in the analytical ultracentrifuge. From sedimentation-equilibrium experiments a mol. wt. of about 376000 was determined for the native enzyme and 47300 for the enzyme in guanidinium chloride. From these data and measurements of electron micrographs, we have concluded that glutamine synthetase from nodule cytosol consists of eight subunits arranged in two sets of planar tetramers which form a cubical configuration with dimensions of about 10 nm (100 A) across each side. Glutamine synthetase from nodule cytosol has a higher glycine and proline content and a lower content of phenylalanine than the glutamine synthetase that has been prepared from pea seed. The cytosol enzyme contains four half-cystine molecules per subunit, which is in contrast with two reported for the enzyme from pea seed. Enzyme activity is striking influenced by the relative proportion of Mg2+ and Mn2+ in the assay medium. Activity is inhibited by feedback inhibitors and is influenced by energy charge. | [
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PMID:8036 | Activities of citrate synthase and NAD+-linked and NADP+-linked isocitrate dehydrogenase in muscle from vertebrates and invertebrates. | 1. The activities of citrate synthase, NAD+-linked and NADP+-linked isocitrate dehydrogenase were measured in muscles from a large number of animals, in order to provide some indication of the importance of the citric acid cycle in these muscles. According to the differences in enzyme activities, the muscles can be divided into three classes. First, in a number of both vertebrate and invertebrate muscles, the activities of all three enzymes are very low. It is suggested that either the muscles use energy at a very low rate or they rely largely on anaerobic glycolysis for higher rates of energy formation. Second, most insect flight muscles contain high activities of citrate synthase and NAD+-linked isocitrate dehydrogenase, but the activities of the NADP+-linked enzyme are very low. The high activities indicate the dependence of insect flight on energy generated via the citric acid cycle. The flight muscles of the beetles investigated contain high activities of both isocitrate dehydrogenases. Third, other muscles of both vertebrates and invertebrates contain high activities of citrate synthase and NADP+-liniked isocitrate dehydrogenase. Many, if not all, of these muscles are capable of sustained periods of mechanical activity (e.g. heart muscle, pectoral muscles of some birds). Consequently, to support this activity fuel must be supplied continually to the muscle via the circulatory system which, in most animals, also transports oxygen so that energy can be generated by complete oxidation of the fuel. It is suggested that the low activities of NAD+-linked isocitrate dehydrogenase in these muscles may be involved in oxidation of isocitrate in the cycle when the muscles are at rest. 2. A comparison of the maximal activities of the enzymes with the maximal flux through the cycle suggests that, in insect flight muscle, NAD+-linked isocitrate dehydrogenase catalyses a non-equilibrium reaction and citrate synthease catalyses a near-equilibrium reaction. In other muscles, the enzyme-activity data suggest that both citrate synthase and the isocitrate dehydrogenase reactions are near-equilibrium. | Activities of citrate synthase and NAD+-linked and NADP+-linked isocitrate dehydrogenase in muscle from vertebrates and invertebrates. 1. The activities of citrate synthase, NAD+-linked and NADP+-linked isocitrate dehydrogenase were measured in muscles from a large number of animals, in order to provide some indication of the importance of the citric acid cycle in these muscles. According to the differences in enzyme activities, the muscles can be divided into three classes. First, in a number of both vertebrate and invertebrate muscles, the activities of all three enzymes are very low. It is suggested that either the muscles use energy at a very low rate or they rely largely on anaerobic glycolysis for higher rates of energy formation. Second, most insect flight muscles contain high activities of citrate synthase and NAD+-linked isocitrate dehydrogenase, but the activities of the NADP+-linked enzyme are very low. The high activities indicate the dependence of insect flight on energy generated via the citric acid cycle. The flight muscles of the beetles investigated contain high activities of both isocitrate dehydrogenases. Third, other muscles of both vertebrates and invertebrates contain high activities of citrate synthase and NADP+-liniked isocitrate dehydrogenase. Many, if not all, of these muscles are capable of sustained periods of mechanical activity (e.g. heart muscle, pectoral muscles of some birds). Consequently, to support this activity fuel must be supplied continually to the muscle via the circulatory system which, in most animals, also transports oxygen so that energy can be generated by complete oxidation of the fuel. It is suggested that the low activities of NAD+-linked isocitrate dehydrogenase in these muscles may be involved in oxidation of isocitrate in the cycle when the muscles are at rest. 2. A comparison of the maximal activities of the enzymes with the maximal flux through the cycle suggests that, in insect flight muscle, NAD+-linked isocitrate dehydrogenase catalyses a non-equilibrium reaction and citrate synthease catalyses a near-equilibrium reaction. In other muscles, the enzyme-activity data suggest that both citrate synthase and the isocitrate dehydrogenase reactions are near-equilibrium. | [
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PMID:8037 | Autophagy-related changes of arylsulphatases A and B in rat liver lysosomes. | The total arylsulphatase activity and the relative activities of lysosomal arylsulphatases A and B were measured in the liver of control rats and rats subjected to treatments that provoke hepatic autophagocytosis. The total liver arylsulphatase activities were increased in starved and starved glucagon-treated rats, but not in sham-operated and hepatectomized rats. Arylsulphatases A and B in the mitochondrial-lysosomal (M-L) fraction were separated by polyacrylamide-gel electrophoresis at pH 8.8; they were made visible by incubating the gels with p-nitrocatechol sulphate as substrate, and measured by quantitative densitometry. In untreated controls, arylsulphatases A and B comprised 41.4 +/- 0.5% and 58.6 +/- 0.5% of the total arylsulphatase activity respectively; the arylsulphatase A/arylsulphatase B activity ratio was 0.71. All experimental treatments produced a significant decrease in the percentage of lysosomal arylsulphatase present as the A form and an increase in that present as the B form, and the activity ratio of arylsulphatase A/arylsulphatase B declined. The magnitude of these changes increased in the following direction: starvation for 24h=sham hepatectomy less than glucagon + starvation less than subtotal hepatectomy. These results indicate that the arylsulphatase A/arylsulphatase B activity ratio in liver lysosomes of normal rats is maintained within rather narrow limits, and this ratio declines during enhanced autophagocytosis. These findings, together with observations that suggest that arylsulphatase B may be a partially degraded form of arylsulphatase A, are consistent with the view that the A form is more rapidly converted into the B form during autophagy, owing to the digestive activity of the other lysosomal hydrolases present in autophagic vacuoles. | Autophagy-related changes of arylsulphatases A and B in rat liver lysosomes. The total arylsulphatase activity and the relative activities of lysosomal arylsulphatases A and B were measured in the liver of control rats and rats subjected to treatments that provoke hepatic autophagocytosis. The total liver arylsulphatase activities were increased in starved and starved glucagon-treated rats, but not in sham-operated and hepatectomized rats. Arylsulphatases A and B in the mitochondrial-lysosomal (M-L) fraction were separated by polyacrylamide-gel electrophoresis at pH 8.8; they were made visible by incubating the gels with p-nitrocatechol sulphate as substrate, and measured by quantitative densitometry. In untreated controls, arylsulphatases A and B comprised 41.4 +/- 0.5% and 58.6 +/- 0.5% of the total arylsulphatase activity respectively; the arylsulphatase A/arylsulphatase B activity ratio was 0.71. All experimental treatments produced a significant decrease in the percentage of lysosomal arylsulphatase present as the A form and an increase in that present as the B form, and the activity ratio of arylsulphatase A/arylsulphatase B declined. The magnitude of these changes increased in the following direction: starvation for 24h=sham hepatectomy less than glucagon + starvation less than subtotal hepatectomy. These results indicate that the arylsulphatase A/arylsulphatase B activity ratio in liver lysosomes of normal rats is maintained within rather narrow limits, and this ratio declines during enhanced autophagocytosis. These findings, together with observations that suggest that arylsulphatase B may be a partially degraded form of arylsulphatase A, are consistent with the view that the A form is more rapidly converted into the B form during autophagy, owing to the digestive activity of the other lysosomal hydrolases present in autophagic vacuoles. | [
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PMID:8038 | Hydrogen-ion titration studies on erythrocyte membranes. | 1. H+ titration was used to detect the presence of ionizable groups on human erythrocyte plasma membranes. Between pH2.9 and 11.3, two significant peaks of H+ association/dissociation occur in the differential from of the titration curve, one at pH3. 1. And the other at pH10.3. 2. After disruption of membrane structure by exposure to high pH or by the addition of sodium dodecyl sulphate, maxima of H+ association/dissociation were seen at pH3.1,4.3,6.5,10.3 and 10.7. 3. Spectrophotometric assay and selective chemical treatments were used to identify several of the titratable residues. 4. The degree of eleectrostatic interaction between titratable charged groups was investigated by comparing the titration characteristics of the membranes before and after modification of membrane structure. | Hydrogen-ion titration studies on erythrocyte membranes. 1. H+ titration was used to detect the presence of ionizable groups on human erythrocyte plasma membranes. Between pH2.9 and 11.3, two significant peaks of H+ association/dissociation occur in the differential from of the titration curve, one at pH3. 1. And the other at pH10.3. 2. After disruption of membrane structure by exposure to high pH or by the addition of sodium dodecyl sulphate, maxima of H+ association/dissociation were seen at pH3.1,4.3,6.5,10.3 and 10.7. 3. Spectrophotometric assay and selective chemical treatments were used to identify several of the titratable residues. 4. The degree of eleectrostatic interaction between titratable charged groups was investigated by comparing the titration characteristics of the membranes before and after modification of membrane structure. | [
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PMID:8039 | The route of secretion of procollagen. The influence of alphaalpha'-bipyridyl, colchicine and antimycin A on the secretory process in embryonic-chick tendon and cartilage cells. | I. Embryonic-chick tendon cells were pulse-labelled for 4 min with [14C]proline and the 14C-labelled polypeptides were chased with unlabelled proline for up to 30 min. Isolation of subcellular fractions during the chase period and their subsequent analysis for bacterial collagenase-susceptible 14C-labelled peptides demonstrated the transfer of procollagen polypeptides from rough to smooth microsomal fractions and thence to the extracellular medium. Parallel analyses of Golgi-enriched fractions indicated the involvement of this organelle in the secretory pathway of procollagen. Sodium dodecylsulphate/polyacrylamide-gel electrophoresis of the 14C-labelled polypeptides present in the Golgi-enriched fractions demonstrated that the procollagen polypeptides were all present as disulphide-linked pro-gamma components. 2. When similar kinetic studies of the intracellular transport of procollagen were conducted with embryonic-chick cartilage cells almost identical results were obtained, but the rate of translocation of cartilage procollagen was significantly slower than that observed for tendon procollagen. 3. When hydroxylation of procollagen polypeptides was inhibited by alphaalpha'-bipyridyl, the nascent polypeptides accumulated in the rough microsomal fraction. 4. When cells were pulse-labelled for 4min with [14C)proline and the label was chased in the presence of colchicine, secretion of procollagen was inhibited and an intracellular accumulation of procollagen 14C-labelled polypeptides was observed in the Golgi-enriched fractions. 5. The energy-dependence of the intracellular transport of procollagen was demonstrated in experiments in which antimycin A was found to inhibit the transfer of procollagen polypeptides from rough to smooth endoplasmic reticulum. 6. It is concluded that procollagen follows the classical route of secretion taken by other extracellular proteins. | The route of secretion of procollagen. The influence of alphaalpha'-bipyridyl, colchicine and antimycin A on the secretory process in embryonic-chick tendon and cartilage cells. I. Embryonic-chick tendon cells were pulse-labelled for 4 min with [14C]proline and the 14C-labelled polypeptides were chased with unlabelled proline for up to 30 min. Isolation of subcellular fractions during the chase period and their subsequent analysis for bacterial collagenase-susceptible 14C-labelled peptides demonstrated the transfer of procollagen polypeptides from rough to smooth microsomal fractions and thence to the extracellular medium. Parallel analyses of Golgi-enriched fractions indicated the involvement of this organelle in the secretory pathway of procollagen. Sodium dodecylsulphate/polyacrylamide-gel electrophoresis of the 14C-labelled polypeptides present in the Golgi-enriched fractions demonstrated that the procollagen polypeptides were all present as disulphide-linked pro-gamma components. 2. When similar kinetic studies of the intracellular transport of procollagen were conducted with embryonic-chick cartilage cells almost identical results were obtained, but the rate of translocation of cartilage procollagen was significantly slower than that observed for tendon procollagen. 3. When hydroxylation of procollagen polypeptides was inhibited by alphaalpha'-bipyridyl, the nascent polypeptides accumulated in the rough microsomal fraction. 4. When cells were pulse-labelled for 4min with [14C)proline and the label was chased in the presence of colchicine, secretion of procollagen was inhibited and an intracellular accumulation of procollagen 14C-labelled polypeptides was observed in the Golgi-enriched fractions. 5. The energy-dependence of the intracellular transport of procollagen was demonstrated in experiments in which antimycin A was found to inhibit the transfer of procollagen polypeptides from rough to smooth endoplasmic reticulum. 6. It is concluded that procollagen follows the classical route of secretion taken by other extracellular proteins. | [
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PMID:8040 | The effects of proteolytic digestion by trypsin on the structure and catalytic properties of reduced nicotinamide-adenine dinucleotide dehydrogenase from bovine heart mitochondria. | 1. At 21 degrees C incubation of NADH-ubiquinone-1 reductase (Complex 1) with trypsin caused selective inhibition of nicotinamide nucleotide transhydrogenase activity. The reduction of K3Fe(CN)6 by NADH or NADPH was unaffected, but a slow decrease in the rate of reduction of ubiquinone-1 by NADH was observed. 2. The pH-dependence of nicotinamide nucleotide transhydrogenase activity differed in Complex I and trypsin-treated Complex I. The trypsin-labile activity had a pH optimum of approx. 6.5, whereas the trypsin-resistant activity had a pH optimum of approx. 5.5 or less. 3. The trypsinlabile transhydrogenase activity was specifically inhibited by butanedione or phenylglyoxal and was identified with the enzyme catalysing energy-linked transhydrogenase activity in submitochondrial particles. 4. Polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate revealed that trypsin caused degradation of a polypeptide of mol.wt 20500 in parallel with the loss of transhydrogenase activity. 5. At 30 degrees C and higher trypsin concentrations, the rate of reduction of K3Fe(CN)6 by NADH or NADPH slowly decreased. Increased lability of NADH-K3Fe(CN)6 reductase activity to trypsin was observed when the endogenous phospholipid of Complex I was depleted by detergent or phospholipase A treatment. 6. Polyacrylamide-gel electrophoresis indicated that removal of phospholipid allowed much more extensive degradation of constituent polypeptides by trypsin. The subunits of the low-molecular-weight (type II) dehydrogenase (53000 and 26000 mol.wt.) were, however, relatively resistant to trypsin even in phospholipid-depleted preparations. | The effects of proteolytic digestion by trypsin on the structure and catalytic properties of reduced nicotinamide-adenine dinucleotide dehydrogenase from bovine heart mitochondria. 1. At 21 degrees C incubation of NADH-ubiquinone-1 reductase (Complex 1) with trypsin caused selective inhibition of nicotinamide nucleotide transhydrogenase activity. The reduction of K3Fe(CN)6 by NADH or NADPH was unaffected, but a slow decrease in the rate of reduction of ubiquinone-1 by NADH was observed. 2. The pH-dependence of nicotinamide nucleotide transhydrogenase activity differed in Complex I and trypsin-treated Complex I. The trypsin-labile activity had a pH optimum of approx. 6.5, whereas the trypsin-resistant activity had a pH optimum of approx. 5.5 or less. 3. The trypsinlabile transhydrogenase activity was specifically inhibited by butanedione or phenylglyoxal and was identified with the enzyme catalysing energy-linked transhydrogenase activity in submitochondrial particles. 4. Polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate revealed that trypsin caused degradation of a polypeptide of mol.wt 20500 in parallel with the loss of transhydrogenase activity. 5. At 30 degrees C and higher trypsin concentrations, the rate of reduction of K3Fe(CN)6 by NADH or NADPH slowly decreased. Increased lability of NADH-K3Fe(CN)6 reductase activity to trypsin was observed when the endogenous phospholipid of Complex I was depleted by detergent or phospholipase A treatment. 6. Polyacrylamide-gel electrophoresis indicated that removal of phospholipid allowed much more extensive degradation of constituent polypeptides by trypsin. The subunits of the low-molecular-weight (type II) dehydrogenase (53000 and 26000 mol.wt.) were, however, relatively resistant to trypsin even in phospholipid-depleted preparations. | [
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PMID:8041 | The regulation of rat liver tryptophan pyrrolase activity by reduced nicotinamide-adenine dinucleotide (phosphate). Experiments with glucose and nicotinamide. | 1. Chronic administration of glucose or nicotinamide in drinking water inhibits the activity of rat liver tryptophan pyrrolase, and subsequent withdrawal causes an enhancement. The enzyme activity is also inhibited by administration in drinking water of sucrose, but not fructose, which is capable of preventing the glucose effect. 2. The inhibition by glucose or nictinamide is not due to a defective apoenzyme synthesis nor a decreased cofactor availability. 3. The inhibition by nicotinamide is reversed by regeneration of liver NAD+ and NADP+ in vivo by administration of fructose, pyruvate or phenazine methosulphate. Inhibition by glucose is also reversed by the above agents and by NH4Cl. Reversal of inhibition by glucose or nicotinamide is also achieved in vitro by addition of NAD+ or NADP+. 4. Glucose or nicotinamide increases liver [NADPH]. [NADP+] is also increased by nicotinamide. [NADPH] is also increased by sucrose, but not by fructose, which prevents the glucose effect. Phenazine methosulphate prevents the increase in [NADPH] caused by both glucose and nicotinamide. 5. It is suggested that the inhibition of tryptophan pyrrolase activity by glucose or nicotinamide is mediated by both NADPH and NADH. | The regulation of rat liver tryptophan pyrrolase activity by reduced nicotinamide-adenine dinucleotide (phosphate). Experiments with glucose and nicotinamide. 1. Chronic administration of glucose or nicotinamide in drinking water inhibits the activity of rat liver tryptophan pyrrolase, and subsequent withdrawal causes an enhancement. The enzyme activity is also inhibited by administration in drinking water of sucrose, but not fructose, which is capable of preventing the glucose effect. 2. The inhibition by glucose or nictinamide is not due to a defective apoenzyme synthesis nor a decreased cofactor availability. 3. The inhibition by nicotinamide is reversed by regeneration of liver NAD+ and NADP+ in vivo by administration of fructose, pyruvate or phenazine methosulphate. Inhibition by glucose is also reversed by the above agents and by NH4Cl. Reversal of inhibition by glucose or nicotinamide is also achieved in vitro by addition of NAD+ or NADP+. 4. Glucose or nicotinamide increases liver [NADPH]. [NADP+] is also increased by nicotinamide. [NADPH] is also increased by sucrose, but not by fructose, which prevents the glucose effect. Phenazine methosulphate prevents the increase in [NADPH] caused by both glucose and nicotinamide. 5. It is suggested that the inhibition of tryptophan pyrrolase activity by glucose or nicotinamide is mediated by both NADPH and NADH. | [
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PMID:8056 | Basic ethers of cyclohexylphenols with beta-blocking activity: synthesis and pharmacological study of exaprolol. | The paper describes the preparation and study of the pharmacological properties of a series of derivatives with the general formula. (see article) where R is a cyclohexyl or cyclohexenyl radical and R1 is a hydrogen, methyl or cyclohexyl. The compounds with a single cycloaliphatic radical ortho to the basic chain, and in particular the one with a cyclohexyl (exaprolol), were found to be particularly active in blocking the beta-adrenergic receptors, as antiarrhythmics and local anesthetics, while the introduction of a second radical or the shift of the cycloaliphatic radical to meta or para position caused the said pharmacological activities to disappear almost entirely, with the exception of the local anesthetic action. | Basic ethers of cyclohexylphenols with beta-blocking activity: synthesis and pharmacological study of exaprolol. The paper describes the preparation and study of the pharmacological properties of a series of derivatives with the general formula. (see article) where R is a cyclohexyl or cyclohexenyl radical and R1 is a hydrogen, methyl or cyclohexyl. The compounds with a single cycloaliphatic radical ortho to the basic chain, and in particular the one with a cyclohexyl (exaprolol), were found to be particularly active in blocking the beta-adrenergic receptors, as antiarrhythmics and local anesthetics, while the introduction of a second radical or the shift of the cycloaliphatic radical to meta or para position caused the said pharmacological activities to disappear almost entirely, with the exception of the local anesthetic action. | [
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PMID:8057 | General pharmacological properties of a new non-opiate antitussive: zipeprol (3024 CERM). I. Action on respiratory function and acute toxicity. | 1-(2-Methoxy-2-phenyl)-ethyl-4-(2-hydroxy-3-methoxy-3-phenyl)-propyl-iperazine-dihydrochloride (zipeprol, Respilene) is a substance of non-phenanthrenic chemical structure. In the cat, it antagonised cough induced by stimulation of the superior laryngeal nerve or by direct mechanical excitation of the sensitive tracheo-bronchial receptors. The efficacy of zipeprol after enteral administration made it possible both to establish good intestinal absorption and to rank it favourably in relation to several major antitussive reference products; codeine, codethyline, dextromethorphan, diphenhydramine and pentoxyverine. The activity of zipeprol was superior or equal to that of all these substances, excdept codeine. The antitussive properties appeared to be due to a central action. Other properties have been demonstrated which suggest at least a supplementary mechanism in the inhibition of cough, in addition to the central action. These consisted of slight antihistamine and anticholinergic properties, marked local-anesthetic potency and bronchospasmolytic activity. This latter property was demonstrated by the inhibition of histamine and serotonin induced bronchospasm in the guinea-pig. In vitro, using human sputum, zipeprol had a mucolytic action, shown by a decrease in sputum vis viscosity and lysis of DNA and AMPS fibrils. In the dog, at high doses, zipeprol unlike codeine, did not inhibit central stimulation of respiration by hypercapnia, in addition no modification of ventilatory dynamics or blood gases was seen. On the basis of these results, zipeprol can be considered as possessing no respiratory depressant effect even in the upper ranges of its antitussive doses. | General pharmacological properties of a new non-opiate antitussive: zipeprol (3024 CERM). I. Action on respiratory function and acute toxicity. 1-(2-Methoxy-2-phenyl)-ethyl-4-(2-hydroxy-3-methoxy-3-phenyl)-propyl-iperazine-dihydrochloride (zipeprol, Respilene) is a substance of non-phenanthrenic chemical structure. In the cat, it antagonised cough induced by stimulation of the superior laryngeal nerve or by direct mechanical excitation of the sensitive tracheo-bronchial receptors. The efficacy of zipeprol after enteral administration made it possible both to establish good intestinal absorption and to rank it favourably in relation to several major antitussive reference products; codeine, codethyline, dextromethorphan, diphenhydramine and pentoxyverine. The activity of zipeprol was superior or equal to that of all these substances, excdept codeine. The antitussive properties appeared to be due to a central action. Other properties have been demonstrated which suggest at least a supplementary mechanism in the inhibition of cough, in addition to the central action. These consisted of slight antihistamine and anticholinergic properties, marked local-anesthetic potency and bronchospasmolytic activity. This latter property was demonstrated by the inhibition of histamine and serotonin induced bronchospasm in the guinea-pig. In vitro, using human sputum, zipeprol had a mucolytic action, shown by a decrease in sputum vis viscosity and lysis of DNA and AMPS fibrils. In the dog, at high doses, zipeprol unlike codeine, did not inhibit central stimulation of respiration by hypercapnia, in addition no modification of ventilatory dynamics or blood gases was seen. On the basis of these results, zipeprol can be considered as possessing no respiratory depressant effect even in the upper ranges of its antitussive doses. | [
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PMID:8058 | [Bronchospasmolytic activity of aqueous aerosols in anesthetized dogs (author's transl)]. | A modification of the method of Konzett and Rössler is described. This modification allows the administration of atomised drug solutions to anaesthetised dogs. Information is thus obtained on the bronchodilatory activity of a drug and the onset and duration of effect. Two anticholinergics (atropinsulfate and ipratropiumbromide) and three beta-adrenoceptor stimulants (isoprenaline, orciprenaline and salbutamol) were investigated. After local administration as aerosols the anticholinergics proved to be as potent or even more so than the beta-adrenoceptor stimulants. | [Bronchospasmolytic activity of aqueous aerosols in anesthetized dogs (author's transl)]. A modification of the method of Konzett and Rössler is described. This modification allows the administration of atomised drug solutions to anaesthetised dogs. Information is thus obtained on the bronchodilatory activity of a drug and the onset and duration of effect. Two anticholinergics (atropinsulfate and ipratropiumbromide) and three beta-adrenoceptor stimulants (isoprenaline, orciprenaline and salbutamol) were investigated. After local administration as aerosols the anticholinergics proved to be as potent or even more so than the beta-adrenoceptor stimulants. | [
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PMID:8059 | Actions of narcotics on brain dopamine metabolism and their relevance for "psychomotor" effects. | A review is given about the effects of narcotic analgesics, particularly of morphine, on the dopamine metabolism in the corpus striatum and about the relations of these effects to motility and "psychomotor" phenomena. In rats, acute doses of morphine decrease the dopaminergic neurotransmission in brain, without blocking postsynaptic dopamine receptors. Chronic treatment of rats with morphine reverses these acute effects of morphine and induces symptoms of an increased dopaminergic neurotransmission in brain. In mice and cats on the other hand, acute doses of morphine apparently increase dopaminergic neurotransmission. The effects of morphine on striatal dopamine metabolism seem to be a model well suited to study opioid-specific effects on a cellular level. Furthermore, they might also be responsible for some narcotic-specific effects on behaviour observed in animals and man. | Actions of narcotics on brain dopamine metabolism and their relevance for "psychomotor" effects. A review is given about the effects of narcotic analgesics, particularly of morphine, on the dopamine metabolism in the corpus striatum and about the relations of these effects to motility and "psychomotor" phenomena. In rats, acute doses of morphine decrease the dopaminergic neurotransmission in brain, without blocking postsynaptic dopamine receptors. Chronic treatment of rats with morphine reverses these acute effects of morphine and induces symptoms of an increased dopaminergic neurotransmission in brain. In mice and cats on the other hand, acute doses of morphine apparently increase dopaminergic neurotransmission. The effects of morphine on striatal dopamine metabolism seem to be a model well suited to study opioid-specific effects on a cellular level. Furthermore, they might also be responsible for some narcotic-specific effects on behaviour observed in animals and man. | [
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PMID:8060 | Metabolic deacetylation: in vitro and in vivo studies in man, rat, dog and rhesus monkey with isomeric tetrahydroisoquinolyl derivatives of 3,4-dimethylbenzyl acetate. | The pharmacological activities of a racemic mixture of tetrahydroisoquinolyl derivatives of 3,4-dimethylbenzyl acetate, (+/-) Ro 03-4661 and the corresponding resolved isomers (+) Ro 03-4661 and (--) Ro 03-4661 have been studied in rat and rhesus monkey. The racemate and the (--) isomer showed narcotic analgesic activity by the oral route in both species. Drug metabolism studies indicated that the activity was probably due to metabolic Deacetylation to the corresponding carbinol Ro 03-4632. deacetylation did not occur in the dog, but was observed in the rat and rhesus monkey. In both these species the hydrolysis was more extensive after oral than parenteral administration. In vitro studies also showed considerable species variation in the ability of blood and tissue esterases to hydrolyse the different stereochemical isomers. | Metabolic deacetylation: in vitro and in vivo studies in man, rat, dog and rhesus monkey with isomeric tetrahydroisoquinolyl derivatives of 3,4-dimethylbenzyl acetate. The pharmacological activities of a racemic mixture of tetrahydroisoquinolyl derivatives of 3,4-dimethylbenzyl acetate, (+/-) Ro 03-4661 and the corresponding resolved isomers (+) Ro 03-4661 and (--) Ro 03-4661 have been studied in rat and rhesus monkey. The racemate and the (--) isomer showed narcotic analgesic activity by the oral route in both species. Drug metabolism studies indicated that the activity was probably due to metabolic Deacetylation to the corresponding carbinol Ro 03-4632. deacetylation did not occur in the dog, but was observed in the rat and rhesus monkey. In both these species the hydrolysis was more extensive after oral than parenteral administration. In vitro studies also showed considerable species variation in the ability of blood and tissue esterases to hydrolyse the different stereochemical isomers. | [
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PMID:8061 | Purification and characterization of lipoprotein lipase from human heart. | Human heart lipoprotein lipase was purified by affinity chromatography on heparin-Sepharose 4B. When crude extracts of heart acetone powder were applied to columsn, about 40% of total lipase activity was bound to the gel and then eluted with 1.5 M NaCl. At this stage the eluted enzyme was purified 1900-fold. Disc gel electrophoresis yielded a single protein band corresponding with lipolytic activity. Minimum molecular weight of the protein was 60,000 as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The purified enzyme was highly unstable; however, its activity could be partially stabilized at --20C by bovine serum albumin, glycerol, or ethylene glycol. The activity of the purified enzyme (i) had a pH optimum between 7.8 and 8.0; (ii) required serum for full enzymatic activity; apoC-II could be substituted for serum; (iii) was inhibited by by apoC-I in the presence of activated substrate; (iv) was markedly inhibited by NaCl; and (v) was stimulated by heparin. | Purification and characterization of lipoprotein lipase from human heart. Human heart lipoprotein lipase was purified by affinity chromatography on heparin-Sepharose 4B. When crude extracts of heart acetone powder were applied to columsn, about 40% of total lipase activity was bound to the gel and then eluted with 1.5 M NaCl. At this stage the eluted enzyme was purified 1900-fold. Disc gel electrophoresis yielded a single protein band corresponding with lipolytic activity. Minimum molecular weight of the protein was 60,000 as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The purified enzyme was highly unstable; however, its activity could be partially stabilized at --20C by bovine serum albumin, glycerol, or ethylene glycol. The activity of the purified enzyme (i) had a pH optimum between 7.8 and 8.0; (ii) required serum for full enzymatic activity; apoC-II could be substituted for serum; (iii) was inhibited by by apoC-I in the presence of activated substrate; (iv) was markedly inhibited by NaCl; and (v) was stimulated by heparin. | [
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PMID:8062 | Neurogenic hypercholesterolemia: influence of autonomic drugs. | Eleven substances capable of either augmenting or depleting the alpha- and - beta-adrenergic capacities of the autonomic nervous system were administered to rats exhibiting hypothalamic hypercholesterolemia and to normal controls. Only the beta-adrenergic blocking agents propranolol and possibly 6-OH dopamine were observed to alter (raise) the serum cholesterol concentration, and this occurred in both experimental and control animals. Neither atropine, nor the serotonin-depleting agent, rho-chlorophenylalanine, nor the serotonin-antagonist cyproheptadine, were observed to alter serum cholesterol level. Such absence of effect was also noted with metaraminol, phenoxybenzamine, isoproterenol, epinephrine, reserpine, and alpha-methyl tyrosine. | Neurogenic hypercholesterolemia: influence of autonomic drugs. Eleven substances capable of either augmenting or depleting the alpha- and - beta-adrenergic capacities of the autonomic nervous system were administered to rats exhibiting hypothalamic hypercholesterolemia and to normal controls. Only the beta-adrenergic blocking agents propranolol and possibly 6-OH dopamine were observed to alter (raise) the serum cholesterol concentration, and this occurred in both experimental and control animals. Neither atropine, nor the serotonin-depleting agent, rho-chlorophenylalanine, nor the serotonin-antagonist cyproheptadine, were observed to alter serum cholesterol level. Such absence of effect was also noted with metaraminol, phenoxybenzamine, isoproterenol, epinephrine, reserpine, and alpha-methyl tyrosine. | [
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PMID:8063 | Effects of some benzodiazepine derivatives on Triton WR-1339-Induced hyperlipidaemia in rats. | Oral administration of the benzodiazepines (diazepam, lorazepam, chlordiazepoxide, bipotassium chlorazepate) in Triton WR-1339-induced (200 mg/kg, blood collection 18 h later) hyperlipidaemia in rats elicited marked decrease of serum total lipids, total cholesterol and triglyceride levels, and alterations in free fatty acid and free glycerol content. The optimal doses for diazepam, lorazepam, chlordiazepoxide and bipotassium chlorazepate were estimated to be 5 mg/kg. Other benzodiazepines, namely oxazepam, medazepam and nitrazepam, elicited only minor changes in serum lipids levels, while with grandaxin no change was observed. The optimal doses of diazepam and lorazepam brought about the same changes in serum lipid content as did clofibrate (90 mg/kg, p. o.). If diazepam, lorazepam, chlordiazepoxide and bipotassium chlorazepate were administered in doses of 5 mg/kg in Triton WR-1339-treated rats (blood collection taken 3 h later), a significant decrease of total lipids and triglyceride levels was observed. The free glycerol level only altered after the administration of chlordiazepoxide, which brought about a significant reduction. | Effects of some benzodiazepine derivatives on Triton WR-1339-Induced hyperlipidaemia in rats. Oral administration of the benzodiazepines (diazepam, lorazepam, chlordiazepoxide, bipotassium chlorazepate) in Triton WR-1339-induced (200 mg/kg, blood collection 18 h later) hyperlipidaemia in rats elicited marked decrease of serum total lipids, total cholesterol and triglyceride levels, and alterations in free fatty acid and free glycerol content. The optimal doses for diazepam, lorazepam, chlordiazepoxide and bipotassium chlorazepate were estimated to be 5 mg/kg. Other benzodiazepines, namely oxazepam, medazepam and nitrazepam, elicited only minor changes in serum lipids levels, while with grandaxin no change was observed. The optimal doses of diazepam and lorazepam brought about the same changes in serum lipid content as did clofibrate (90 mg/kg, p. o.). If diazepam, lorazepam, chlordiazepoxide and bipotassium chlorazepate were administered in doses of 5 mg/kg in Triton WR-1339-treated rats (blood collection taken 3 h later), a significant decrease of total lipids and triglyceride levels was observed. The free glycerol level only altered after the administration of chlordiazepoxide, which brought about a significant reduction. | [
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PMID:8068 | Clinical studies of induction agents XLIII: Flunitrazepam. | Flunitrazepam (Ro 5-4200) has been studied as an induction agent in 220 volunteers or patients. It was assumed to be 10 times as potent as diazepam. The maximum soporific effect did not occur until 90-120 s after injection. There was great individual variation in response to flunitrazepam and some patients did not lose consciousness even after receiving 6 mg (approximately 0.1 mg/kg). Opiate premedication enhanced its action, but delayed recovery. There was a dose-related increase in minor respiratory upset with flunitrazepam in unpremedicated patients and a high frequency of arterial hypotension following large doses given to patients who had received opiate premedication. Venous sequelae were no more frequent than after comparable doses of diazepam. Flunitrazepam was not a very satisfactory drug for the induction of anaesthesia, and recovery was too prolonged for routine use. | Clinical studies of induction agents XLIII: Flunitrazepam. Flunitrazepam (Ro 5-4200) has been studied as an induction agent in 220 volunteers or patients. It was assumed to be 10 times as potent as diazepam. The maximum soporific effect did not occur until 90-120 s after injection. There was great individual variation in response to flunitrazepam and some patients did not lose consciousness even after receiving 6 mg (approximately 0.1 mg/kg). Opiate premedication enhanced its action, but delayed recovery. There was a dose-related increase in minor respiratory upset with flunitrazepam in unpremedicated patients and a high frequency of arterial hypotension following large doses given to patients who had received opiate premedication. Venous sequelae were no more frequent than after comparable doses of diazepam. Flunitrazepam was not a very satisfactory drug for the induction of anaesthesia, and recovery was too prolonged for routine use. | [
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PMID:8070 | Salmefamol and Salbutamol in exercise-induced asthma in children. | A double-blind controlled study is reported of salmefamol 200 mug and salbutamol 200 mug administered by aerosol before exercise tests in asthmatic children. Both drugs prevented exercise-induced asthma. There was no significant difference between them. | Salmefamol and Salbutamol in exercise-induced asthma in children. A double-blind controlled study is reported of salmefamol 200 mug and salbutamol 200 mug administered by aerosol before exercise tests in asthmatic children. Both drugs prevented exercise-induced asthma. There was no significant difference between them. | [
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PMID:8071 | The role of divalent cations in epidermolysis. | In experiments designed to elucidate the role of divalent cations in maintaining the integrity of the epidermis, newborn mouse skin was incubated in ethylenediamine tetraacetic acid (EDTA) or ethylene glycol tetraacetic acid (EGTA). In EDTA, epidermolysis occurred and was confirmed by rubbing the specimen to demonstrate that a sheet of epidermis could be split off. After 30 min incubation in 0-01 mol/EDTA the split occurred in the lower granular-upper spinous layer; after 45 min, it was in a spinous-suprabasilar location and at 60 min and later at the dermal-epidermal junction. Ultrastructurally, a clear zone of apparent intracellular oedema occurred along the cell membranes. The split then occurred intracytoplasmally through this clear zone and the adjacent cell membrane was lost. Since incubation in EGTA at pH 7-4 did not result in epidermolysis, we suggest that the removal of magnesium rather than calcium is responsible for epidermolysis. Thirty min after the addition of either calcium or magnesium to EDTA-treated specimens, epidermolysis could no longer be demonstrated. | The role of divalent cations in epidermolysis. In experiments designed to elucidate the role of divalent cations in maintaining the integrity of the epidermis, newborn mouse skin was incubated in ethylenediamine tetraacetic acid (EDTA) or ethylene glycol tetraacetic acid (EGTA). In EDTA, epidermolysis occurred and was confirmed by rubbing the specimen to demonstrate that a sheet of epidermis could be split off. After 30 min incubation in 0-01 mol/EDTA the split occurred in the lower granular-upper spinous layer; after 45 min, it was in a spinous-suprabasilar location and at 60 min and later at the dermal-epidermal junction. Ultrastructurally, a clear zone of apparent intracellular oedema occurred along the cell membranes. The split then occurred intracytoplasmally through this clear zone and the adjacent cell membrane was lost. Since incubation in EGTA at pH 7-4 did not result in epidermolysis, we suggest that the removal of magnesium rather than calcium is responsible for epidermolysis. Thirty min after the addition of either calcium or magnesium to EDTA-treated specimens, epidermolysis could no longer be demonstrated. | [
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PMID:8073 | Planned pregnancy in a renal transplant recipient. | A scheme is described for the investigation and management of a renal transplant recipient before conception, over conception and during the subsequent pregnancy, delivery and puerperium. The patient showed the physiological changes in homeostasis of normal pregnancy. Methods of monitoring maternal, fetal and renal well-being are discussed. The patient was delivered of a live male child and the neonatal problems are described. The puerperium was uneventful. | Planned pregnancy in a renal transplant recipient. A scheme is described for the investigation and management of a renal transplant recipient before conception, over conception and during the subsequent pregnancy, delivery and puerperium. The patient showed the physiological changes in homeostasis of normal pregnancy. Methods of monitoring maternal, fetal and renal well-being are discussed. The patient was delivered of a live male child and the neonatal problems are described. The puerperium was uneventful. | [
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PMID:8074 | Effect of oral 17-beta-oestradiol on the liver in women with intrahepatic cholestasis (hepatosis) during previous pregnancy. | The effects of 17-beta-oestradiol in 14 women who had had intrahepatic cholestasis of pregnancy (hepatosis) were studied. This natural oestrogen was given orally in a daily dose of 5 mg for two 20-day periods with a 7-day interval between courses. The bromsulphthalein (BSP) retention rose during the treatment, but only occasionally to levels above the upper limit of normal. Other conventional liver function tests did not usually become abnormal. In a previous investigation in the same type of patients, we studied the effects of mestranol (a synthetic 17-alkylated oestrogen) in a daily dose of 0.1 mg with the same dose-schedule as the present investigation and 17-beta-oestradiol appeared to impair liver function less than mestranol. It might thus be safer to use 17-beta-oestradiol as a therapeutic agent. | Effect of oral 17-beta-oestradiol on the liver in women with intrahepatic cholestasis (hepatosis) during previous pregnancy. The effects of 17-beta-oestradiol in 14 women who had had intrahepatic cholestasis of pregnancy (hepatosis) were studied. This natural oestrogen was given orally in a daily dose of 5 mg for two 20-day periods with a 7-day interval between courses. The bromsulphthalein (BSP) retention rose during the treatment, but only occasionally to levels above the upper limit of normal. Other conventional liver function tests did not usually become abnormal. In a previous investigation in the same type of patients, we studied the effects of mestranol (a synthetic 17-alkylated oestrogen) in a daily dose of 0.1 mg with the same dose-schedule as the present investigation and 17-beta-oestradiol appeared to impair liver function less than mestranol. It might thus be safer to use 17-beta-oestradiol as a therapeutic agent. | [
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PMID:8076 | Structural elucidation and properties of 8alpha-(N1-histidyl)riboflavin: the flavin component of thiamine dehydrogenase and beta-cyclopiazonate oxidocyclase. | In addition to 8alpha-(N3-histidyl)riboflavin, 8alpha-(N1-histidyl)riboflavin is also formed during the reaction of Nalpha-blocked histidine with 8alpha-bromotetraacetylriboflavin in a yield of 20-25% of the total histidylflavin fraction. The properties of 8alpha-(N1-histidyl)riboflavin are inditical with those of the histidylflavin isolated from thiamine dehydrogenase and beta-cyclopiazonate oxidocyclase but differ from those of 8alpha-(N3-histidyl)riboflavin. These properties include pKa of fluorescence quenching, electrophoretic mobility at pH 5.0, stability to storage, and reduction by NaBH4. Proof for 8alpha substitution is shown by the electron paramagnetic resonance and electron-nuclear double resonance spectra of the cationic semiquinone form, as well as by the proton magnetic resonance spectrum of the oxidized form. The site of histidine substitution by the 8alpha-methylene of the flavin moiety was shown by methylation of the imidazole ring with methyl iodide, cleavage of the methylhistidine-flavin bond by acid hydrolysis at 150 degrees C, and identification of the methylhistidine isomer by electrophoresis. 3-Methylhistidine is the product from the N1-histidylflavin isomer, while 1-methylhistidine is produced from the N3 isomer. The flavin product from reductive Zn cleavage of either isomer has been identified as riboflavin. The compound obtained on acid treatment of 8alpha-(N3-histidyl)riboflavin (previously thought to be the N1 isomer) differs from the parent compound only in the ribityl side chain, since chemical degradation studies show 1-methylhistidine as a product and a flavin product which differs from riboflavin only in mobility in thin-layer chromatography, but not in absorption, fluorescence, and electron paramagnetic resonance spectral properties. Proof that acid modification involves only the ribityl chain has come from the observations that alkaline irradiation of this flavin yields lumiflavin, that the proton magnetic resonance spectrum of the compound differs from that of riboflavin in the region of the ribityl proton resonance, and that its periodate titer is lower than that of authentic riboflavin. The identity of 8alpha-(N1-histidyl)riboflavin with the histidylflavin from thiamine dehydrogenase and beta-cyclopiazonate oxidocyclase shows that both isomeric forms of 8alpha-histidylflavin occur in nature. | Structural elucidation and properties of 8alpha-(N1-histidyl)riboflavin: the flavin component of thiamine dehydrogenase and beta-cyclopiazonate oxidocyclase. In addition to 8alpha-(N3-histidyl)riboflavin, 8alpha-(N1-histidyl)riboflavin is also formed during the reaction of Nalpha-blocked histidine with 8alpha-bromotetraacetylriboflavin in a yield of 20-25% of the total histidylflavin fraction. The properties of 8alpha-(N1-histidyl)riboflavin are inditical with those of the histidylflavin isolated from thiamine dehydrogenase and beta-cyclopiazonate oxidocyclase but differ from those of 8alpha-(N3-histidyl)riboflavin. These properties include pKa of fluorescence quenching, electrophoretic mobility at pH 5.0, stability to storage, and reduction by NaBH4. Proof for 8alpha substitution is shown by the electron paramagnetic resonance and electron-nuclear double resonance spectra of the cationic semiquinone form, as well as by the proton magnetic resonance spectrum of the oxidized form. The site of histidine substitution by the 8alpha-methylene of the flavin moiety was shown by methylation of the imidazole ring with methyl iodide, cleavage of the methylhistidine-flavin bond by acid hydrolysis at 150 degrees C, and identification of the methylhistidine isomer by electrophoresis. 3-Methylhistidine is the product from the N1-histidylflavin isomer, while 1-methylhistidine is produced from the N3 isomer. The flavin product from reductive Zn cleavage of either isomer has been identified as riboflavin. The compound obtained on acid treatment of 8alpha-(N3-histidyl)riboflavin (previously thought to be the N1 isomer) differs from the parent compound only in the ribityl side chain, since chemical degradation studies show 1-methylhistidine as a product and a flavin product which differs from riboflavin only in mobility in thin-layer chromatography, but not in absorption, fluorescence, and electron paramagnetic resonance spectral properties. Proof that acid modification involves only the ribityl chain has come from the observations that alkaline irradiation of this flavin yields lumiflavin, that the proton magnetic resonance spectrum of the compound differs from that of riboflavin in the region of the ribityl proton resonance, and that its periodate titer is lower than that of authentic riboflavin. The identity of 8alpha-(N1-histidyl)riboflavin with the histidylflavin from thiamine dehydrogenase and beta-cyclopiazonate oxidocyclase shows that both isomeric forms of 8alpha-histidylflavin occur in nature. | [
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PMID:8077 | Energetics of primary processes in visula escitation: photocalorimetry of rhodopsin in rod outer segment membranes. | A sensitive technique for the direct calorimetric determination of the energetics of photochemical reactions under low levels of illumination, and its application to the study of primary processes in visula excitation, are described. Enthlpies are reported for various steps in the bleaching of rhodopsin in intact rod outer segment membranes, together with the heats of appropriate model reactions. Protonation changes are also determined calorimetrically by use of buffers with differing heats of proton ionization. Bleaching of rhodopsin is accompanied by significant uptake of heat energy, vastly in excess of the energy required for simple isomerization of the retinal chromophore. Metarhodopsin I formation involves the uptake of about 17 kcal/mol and no net change in proton ionization of the system. Formation of metarhodopsin II requires an additional energy of about 10 kcal/mol and involves the uptake on one hydrogen ion from solution. The energetics of the overall photolysis reaction, rhodopsin leads to opsin + all-trans-retinal, are pH dependent and involve the exposure of an additional titrating group on opsin. This group has a heat of proton ionization of about 12 kcal/mal, characteristic of a primary amine, but a pKa in the region of neutrality. We suggest that this group is the Schiff base lysine of the chromophore binding site of rhodopsin which becomes exposed on photolysis. The low pKa for this active lysine would result in a more stable retinal-opsin linkage, and might be induced by a nearby positively charged group on the protein (either arginine or a second lysine residue). This leads to a model involving intramolecular protonation of the Schiff base nitrogen in the retinal-opsin linkage of rhodopsin, which is consistent with the thermodynamic and spectroscopic properties of the system. We further propose that the metarhodopsin I leads to metarhodopsin II step in the bleaching sequence involves reversible hydrolysis of the Schiff base linkage in the chromophore binding site, and that subsequent steps are the result of migration of the chromophore from this site. | Energetics of primary processes in visula escitation: photocalorimetry of rhodopsin in rod outer segment membranes. A sensitive technique for the direct calorimetric determination of the energetics of photochemical reactions under low levels of illumination, and its application to the study of primary processes in visula excitation, are described. Enthlpies are reported for various steps in the bleaching of rhodopsin in intact rod outer segment membranes, together with the heats of appropriate model reactions. Protonation changes are also determined calorimetrically by use of buffers with differing heats of proton ionization. Bleaching of rhodopsin is accompanied by significant uptake of heat energy, vastly in excess of the energy required for simple isomerization of the retinal chromophore. Metarhodopsin I formation involves the uptake of about 17 kcal/mol and no net change in proton ionization of the system. Formation of metarhodopsin II requires an additional energy of about 10 kcal/mol and involves the uptake on one hydrogen ion from solution. The energetics of the overall photolysis reaction, rhodopsin leads to opsin + all-trans-retinal, are pH dependent and involve the exposure of an additional titrating group on opsin. This group has a heat of proton ionization of about 12 kcal/mal, characteristic of a primary amine, but a pKa in the region of neutrality. We suggest that this group is the Schiff base lysine of the chromophore binding site of rhodopsin which becomes exposed on photolysis. The low pKa for this active lysine would result in a more stable retinal-opsin linkage, and might be induced by a nearby positively charged group on the protein (either arginine or a second lysine residue). This leads to a model involving intramolecular protonation of the Schiff base nitrogen in the retinal-opsin linkage of rhodopsin, which is consistent with the thermodynamic and spectroscopic properties of the system. We further propose that the metarhodopsin I leads to metarhodopsin II step in the bleaching sequence involves reversible hydrolysis of the Schiff base linkage in the chromophore binding site, and that subsequent steps are the result of migration of the chromophore from this site. | [
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PMID:8078 | Characterization of 20S component in tubulin from mammalian brain. | Tubulin from porcine brain, purified by at least two cycles of assembly and disassembly, was characerized at different pH values by sedimentation velocity analysis and turbidimetric measurements. At pH 6.4 the depolymerized material was composed of two major species sedimenting with so20,w values of 6 and 36 and a minor one of 20S. By raising the pH, the amount of the 20S component increased and that of the 36S decreased, whereas that of the 6S component was unaltered. At pH 7.6 the mixture contained 20S and 6S components but hardly any 36S. The 20S species can be separated from the 6S ones by gel filtration on agarose A-15m at pH 7.6. On electron microscopic examination this preparation contains far fewer double rings compared to the material at pH 6.4, but single rings could often be seen. Sodium dodecyl sulfate gel electrophoresis of the 20S and 36S components showed that they consist almost entirely of tubulin and some higher and lower molecular weight fractions. Turbidity measurements showed that the minimal protein concentration necessary for polymerization increases with increasing pH. The turbidity plateau reached at a given pH can be raised by decreasing the pH. From these results it is suggested that the 20S component is an intermediate of the 36S species. The results further indicate the existence of a pH-dependent equilibrium between the 20S species and the 36Soligomers. | Characterization of 20S component in tubulin from mammalian brain. Tubulin from porcine brain, purified by at least two cycles of assembly and disassembly, was characerized at different pH values by sedimentation velocity analysis and turbidimetric measurements. At pH 6.4 the depolymerized material was composed of two major species sedimenting with so20,w values of 6 and 36 and a minor one of 20S. By raising the pH, the amount of the 20S component increased and that of the 36S decreased, whereas that of the 6S component was unaltered. At pH 7.6 the mixture contained 20S and 6S components but hardly any 36S. The 20S species can be separated from the 6S ones by gel filtration on agarose A-15m at pH 7.6. On electron microscopic examination this preparation contains far fewer double rings compared to the material at pH 6.4, but single rings could often be seen. Sodium dodecyl sulfate gel electrophoresis of the 20S and 36S components showed that they consist almost entirely of tubulin and some higher and lower molecular weight fractions. Turbidity measurements showed that the minimal protein concentration necessary for polymerization increases with increasing pH. The turbidity plateau reached at a given pH can be raised by decreasing the pH. From these results it is suggested that the 20S component is an intermediate of the 36S species. The results further indicate the existence of a pH-dependent equilibrium between the 20S species and the 36Soligomers. | [
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PMID:8079 | Histone interactions in solution and susceptibility to denaturation. | Histone interactions in solution may depend upon treatments used for purification. Optical rotatory dispersion and sedimentation-velocity measurements have been made in a reference solvent, before and after exposure to various treatments, to investigate histone susceptibility to irreversible denaturation. Some acid conditions and urea and guanidine solutions may denature. Interaction studies performed on nondenatured histones indicate that the dimer, (H4)(H3), and tetramer, (H4)2(H3)2, dissociate to monomers at low ionic strength. Sedimentation-velocity experiments suggest a model for the (H4)2(H3)2 tetramer, with a compact semispherical center and four protruding amino-terminal regions. Fractions H2a and H2b interact to form the mixed dimer in equilibrium with monomers. Fraction H2a self-associates readily to dimers, tetramers, and octamers, while fraction H1 associates only weakly to form dimers. | Histone interactions in solution and susceptibility to denaturation. Histone interactions in solution may depend upon treatments used for purification. Optical rotatory dispersion and sedimentation-velocity measurements have been made in a reference solvent, before and after exposure to various treatments, to investigate histone susceptibility to irreversible denaturation. Some acid conditions and urea and guanidine solutions may denature. Interaction studies performed on nondenatured histones indicate that the dimer, (H4)(H3), and tetramer, (H4)2(H3)2, dissociate to monomers at low ionic strength. Sedimentation-velocity experiments suggest a model for the (H4)2(H3)2 tetramer, with a compact semispherical center and four protruding amino-terminal regions. Fractions H2a and H2b interact to form the mixed dimer in equilibrium with monomers. Fraction H2a self-associates readily to dimers, tetramers, and octamers, while fraction H1 associates only weakly to form dimers. | [
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PMID:8080 | Structural analysis of O2'-methyl-5-carbamoylmethyluridine, a newly discovered constituent of yeast transfer RNA. | A compound tentatively identified as O2-methyl-5-carboxymethyluridine (cm5Um) was recently isolated in this laboratory from bulk yeast transfer RNA (Gray, M. W. (1975), Can, J. Biochem. 53, 735-746). Alkaline hydrolysis of yeast tRNA releases this nucleoside as part of an alkali-stable dinucleotide, cm5Um-Ap, from which sufficient cm5Um was prepared in the present investigation for a detailed examination of its properties. The ultraviolet absorption spectra and chromatographic and electrophoretic properties of cm5Um were consistent with the proposed structure, which was confirmed by characterization of the base and sugar moieties as 5-carboxymethyluracil and 2-O-methylribose, respectively. Snake venom hydrolysis of yeast tRNA releases cm5Um in the form of a carboxyl-blocked 5'-nucleotide, designated pU-2. Identification of the alkali-labile blocking group in pU-2 as an amide was based on quantitative assay for ammonia released upon acid hydrolysis of the corresponding nucleoside, U-2, and by chromatographic comparison of U-2 with the semisynthetic methyl ester and amide derivatives of cm5Um (mcm5Um and ncm5Um, respectively). Quantitative analysis has indicated that ncm5Um may be confined to a single species of yeast tRNA. In view of the unique localization (the "Wobble" position of the anticodon sequence) and coding properties (pairing with A but not with G) of other cm5U derivatives in transfer RNA, the dinucleotide cm5Um-Ap may be derived from the first two positions of the anticodon sequence of a yeast tRNA species recognizing an NUA codon. This predicts that O2-methyl-5-carbamoylmethyluridine will be found in an isoleucine, leucine, or valine isoacceptor. | Structural analysis of O2'-methyl-5-carbamoylmethyluridine, a newly discovered constituent of yeast transfer RNA. A compound tentatively identified as O2-methyl-5-carboxymethyluridine (cm5Um) was recently isolated in this laboratory from bulk yeast transfer RNA (Gray, M. W. (1975), Can, J. Biochem. 53, 735-746). Alkaline hydrolysis of yeast tRNA releases this nucleoside as part of an alkali-stable dinucleotide, cm5Um-Ap, from which sufficient cm5Um was prepared in the present investigation for a detailed examination of its properties. The ultraviolet absorption spectra and chromatographic and electrophoretic properties of cm5Um were consistent with the proposed structure, which was confirmed by characterization of the base and sugar moieties as 5-carboxymethyluracil and 2-O-methylribose, respectively. Snake venom hydrolysis of yeast tRNA releases cm5Um in the form of a carboxyl-blocked 5'-nucleotide, designated pU-2. Identification of the alkali-labile blocking group in pU-2 as an amide was based on quantitative assay for ammonia released upon acid hydrolysis of the corresponding nucleoside, U-2, and by chromatographic comparison of U-2 with the semisynthetic methyl ester and amide derivatives of cm5Um (mcm5Um and ncm5Um, respectively). Quantitative analysis has indicated that ncm5Um may be confined to a single species of yeast tRNA. In view of the unique localization (the "Wobble" position of the anticodon sequence) and coding properties (pairing with A but not with G) of other cm5U derivatives in transfer RNA, the dinucleotide cm5Um-Ap may be derived from the first two positions of the anticodon sequence of a yeast tRNA species recognizing an NUA codon. This predicts that O2-methyl-5-carbamoylmethyluridine will be found in an isoleucine, leucine, or valine isoacceptor. | [
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PMID:8081 | Mechanisms of electron transfer from sulfite to horseradish peroxidase-hydroperoxide compounds. | Using a rapid-scan spectrophotometer equipped with a stopped-flow apparatus, reactions of sulfite with compounds I and II of two horseradish peroxidase isoenzymes A and C were investigated. The direct two-electron reduction of peroxidase compound I by sulfite occurred at acidic pH but the mechanism gradually changed to the two-step reduction with the intermediate formation of compound II as the pH increased. The pH at which the one- and two-electron changes occurred at the same speed was 4.5 for peroxidase A and 7.7 for peroxidase C. A new peroxidase intermediate was found in the reaction between peroxidase compound II and sulfite. The sulfite compound showed a characteristic absorption band at 850 nm and the optical spectrum was similar to that of isoporphyrins but was quite different from that of sulfhemoproteins. The rate (k) of conversion from the sulfite-compound II complex to the sulfite compound was proportional to the concentration of H+ and the log k vs. pH plot for peroxidase A moved to the acidic side by 1.1 pH unit from that for peroxidase C. | Mechanisms of electron transfer from sulfite to horseradish peroxidase-hydroperoxide compounds. Using a rapid-scan spectrophotometer equipped with a stopped-flow apparatus, reactions of sulfite with compounds I and II of two horseradish peroxidase isoenzymes A and C were investigated. The direct two-electron reduction of peroxidase compound I by sulfite occurred at acidic pH but the mechanism gradually changed to the two-step reduction with the intermediate formation of compound II as the pH increased. The pH at which the one- and two-electron changes occurred at the same speed was 4.5 for peroxidase A and 7.7 for peroxidase C. A new peroxidase intermediate was found in the reaction between peroxidase compound II and sulfite. The sulfite compound showed a characteristic absorption band at 850 nm and the optical spectrum was similar to that of isoporphyrins but was quite different from that of sulfhemoproteins. The rate (k) of conversion from the sulfite-compound II complex to the sulfite compound was proportional to the concentration of H+ and the log k vs. pH plot for peroxidase A moved to the acidic side by 1.1 pH unit from that for peroxidase C. | [
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PMID:8082 | Vinylglycine and proparglyglycine: complementary suicide substrates for L-amino acid oxidase and D-amino acid oxidase. | Proparglyglycine (2-amino-4-pentynoate) and vinylglycine (2-amino-3-butenoate) have been examined as substrates and possible inactivators of two flavo enzymes, D-amino acid oxidase from pig kidney and L-amino acid oxidase from Crotalus adamanteus venom. Vinylglycine is rapidly oxidized by both enzymes but only L-amino acid oxidase is inactivated under assay conditions. The loss of activity probably involves covalent modification of an active site residue rather than the flavin adenine dinucleotide coenzyme and occurs once every 20000 turnovers. We have confirmed the recent observation (Horiike, K, Hishina, Y., Miyake, Y., and Yamano, T. (1975) J, Biochem. (Tokyo), 78, 57) that D-proparglglycine is oxidized with a time-dependent loss of activity by D-amino acid oxidase and have examined some mechanistic aspects of this inactivation, The extent of residual oxidase activity, insensitive to further inactivation, is about 2%, at which point 1.7 labels/subunit have been introduced with propargly[2-14C]glycine as substrate. L-Proparglyclycine is a substrate but not an inactivator of L-amino acid oxidase and the product ahat accumulats in the nonnucleophilic N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffer is acetopyruvate. In the presence of butylamine HCl, a species with lambdaman 317 nm (epsilon = 15 000) accumulates that may be a conjugated eneamine adduct. The same species accumulates from D-amino acid oxidase oxidation of D-propargylglycine prior to inactivation; the inactivated apo D-amino acid oxidase has a new peak at 317 nm that is probably a similar eneamine. A likely inactivating species is 2-keto-3,4-pentadienoate arising from facile rearrangement of the expected initial product 2-keto 4 pentynoate. Vinylglycine and proparglyglycine show inactivation specificity, then, for L-and D-amino acid oxidase, respectively. | Vinylglycine and proparglyglycine: complementary suicide substrates for L-amino acid oxidase and D-amino acid oxidase. Proparglyglycine (2-amino-4-pentynoate) and vinylglycine (2-amino-3-butenoate) have been examined as substrates and possible inactivators of two flavo enzymes, D-amino acid oxidase from pig kidney and L-amino acid oxidase from Crotalus adamanteus venom. Vinylglycine is rapidly oxidized by both enzymes but only L-amino acid oxidase is inactivated under assay conditions. The loss of activity probably involves covalent modification of an active site residue rather than the flavin adenine dinucleotide coenzyme and occurs once every 20000 turnovers. We have confirmed the recent observation (Horiike, K, Hishina, Y., Miyake, Y., and Yamano, T. (1975) J, Biochem. (Tokyo), 78, 57) that D-proparglglycine is oxidized with a time-dependent loss of activity by D-amino acid oxidase and have examined some mechanistic aspects of this inactivation, The extent of residual oxidase activity, insensitive to further inactivation, is about 2%, at which point 1.7 labels/subunit have been introduced with propargly[2-14C]glycine as substrate. L-Proparglyclycine is a substrate but not an inactivator of L-amino acid oxidase and the product ahat accumulats in the nonnucleophilic N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffer is acetopyruvate. In the presence of butylamine HCl, a species with lambdaman 317 nm (epsilon = 15 000) accumulates that may be a conjugated eneamine adduct. The same species accumulates from D-amino acid oxidase oxidation of D-propargylglycine prior to inactivation; the inactivated apo D-amino acid oxidase has a new peak at 317 nm that is probably a similar eneamine. A likely inactivating species is 2-keto-3,4-pentadienoate arising from facile rearrangement of the expected initial product 2-keto 4 pentynoate. Vinylglycine and proparglyglycine show inactivation specificity, then, for L-and D-amino acid oxidase, respectively. | [
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