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1
PMC11277133_p19
PMC11277133
sec[1]/sec[12]/p[0]
2.13. ICAM 1
3.822266
biomedical
Study
[ 0.99951171875, 0.00017058849334716797, 0.00032067298889160156 ]
[ 0.859375, 0.1304931640625, 0.009552001953125, 0.0008325576782226562 ]
ICAM 1 is an endothelial receptor and ligand for leukocyte integrin CD 18. It allows their firm adhesion to the vascular endothelium . There are studies that have demonstrated the increase in soluble ICAM 1 in the context of acute myocardial infarction , but also its overregulation in human cardiomyocytes after infarction. However, this is not detectable in infarcts more recent than one day .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277133_p20
PMC11277133
sec[1]/sec[13]/p[0]
2.14. ILs (Interleukins)
3.951172
biomedical
Study
[ 0.99951171875, 0.00019180774688720703, 0.00023412704467773438 ]
[ 0.8056640625, 0.00957489013671875, 0.1845703125, 0.00048732757568359375 ]
ILs are important inflammatory mediators of acute myocardial infarction, with a role in exacerbating (e.g., IL6) or attenuating (e.g., IL10, IL 15) the acute inflammatory response. They are released from various cells into the bloodstream and interstitium, where they bind to interleukin receptors on cell surfaces, causing cell activation. Some studies have also analyzed the prognosis of acute myocardial infarction according to the level of serum interleukins .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277133_p21
PMC11277133
sec[1]/sec[14]/p[0]
2.15. Monocyte Chemoattractant Protein-1 (MCP 1)
4.074219
biomedical
Study
[ 0.99951171875, 0.00014257431030273438, 0.00015747547149658203 ]
[ 0.99853515625, 0.0004343986511230469, 0.0007791519165039062, 0.0000654458999633789 ]
MCP 1 is a chemokine that induces the recruitment and activation of monocytes, T cells, and NK cells . It is produced by many cells in response to injury factors or exposure to other cytokines, and is also involved in acute myocardial infarction . The antiapoptotic and chemotactic effects of MCP 1 may be mediated by different signaling pathways in the infarcted myocardial area . In the study conducted by Turillazzi et al., a strongly expressed response of MCP-1 was found in the early phase of acute myocardial infarction in the first 0–4 h .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277133_p22
PMC11277133
sec[1]/sec[15]/p[0]
2.16. Tryptase
4.089844
biomedical
Study
[ 0.99951171875, 0.00011080503463745117, 0.00016999244689941406 ]
[ 0.998046875, 0.0009622573852539062, 0.0008540153503417969, 0.00008767843246459961 ]
Tryptase is a protease released by mast cells. The increase in mast cell tryptase plays a central role in the immediate inflammatory and allergic reactions initiated by IgE . It induces fibroblast proliferation, stimulates fibroblast chemotaxis, and regulates the increased production of collagen type I . The same study mentioned above by Turillazzi et al. found a moderate immunohistochemical reaction for tryptase at myocardial infarction sites 0–6 h old, along with CD15, IL-1β, IL-6, TNF-α, IL-8, CD18, ICAM-1, IL-15, and MCP-1 .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277133_p23
PMC11277133
sec[1]/sec[16]/p[0]
2.17. Adiponectin
3.984375
biomedical
Study
[ 0.99951171875, 0.00013494491577148438, 0.00011843442916870117 ]
[ 0.9736328125, 0.00794219970703125, 0.0181121826171875, 0.0003247261047363281 ]
Adiponectin is an adipokine secreted by adipocytes, with anti-inflammatory, antifibrotic, antioxidant, and cardioprotective effects . Experimental studies on animals have proved that in ischemic myocardial lesions, adiponectin accumulates in the heart tissue, where it passes from the vascular compartment. It has a longer half-life in heart tissue than in plasma .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999995
PMC11277133_p24
PMC11277133
sec[1]/sec[17]/p[0]
2.18. Macrophage Migration Inhibitory Factor (MIF)
4.140625
biomedical
Study
[ 0.99951171875, 0.00018846988677978516, 0.00010067224502563477 ]
[ 0.98095703125, 0.012603759765625, 0.005931854248046875, 0.0005087852478027344 ]
MIF is an immunoregulatory cytokine considered a potential biomarker in numerous diseases that also have an inflammatory component . It can be produced by monocytes, macrophages, endocrine, epithelial, and endothelial cells, is stored cytoplasmically, and is rapidly released to stimuli such as microbial products, proliferative signals, and hypoxia . In the case of acute myocardial infarction, MIF increased in myocytes within a few hours, preceding macrophage infiltration in the infarcted area .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277133_p25
PMC11277133
sec[2]/p[0]
3. Discussion
3.992188
biomedical
Review
[ 0.9912109375, 0.005733489990234375, 0.00316619873046875 ]
[ 0.004302978515625, 0.0017995834350585938, 0.9931640625, 0.0005655288696289062 ]
Our review has exposed the main immunohistochemical markers considered over time for diagnosing myocardial infarction more recent than 6 h. They derive from knowledge of the pathophysiological mechanism, and appropriate laboratory elements are required for their determination, as well as complex and expensive forensic analysis . As far as laboratory methods are concerned, there is a need to simultaneously analyze as many markers of recent acute myocardial infarction as possible on the same tissue sample . However, particular aspects of each case must also be considered, such as cardiopulmonary resuscitation, catecholamine injection, drug use (ecstasy, cocaine), agonal artifacts, postmortem interval, autolysis, and pre-existing ischemic events, which can influence immunohistochemical results .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277133_p26
PMC11277133
sec[2]/sec[0]/p[0]
3.1. Agonal Myocardial Immunohistochemical Changes
4.136719
biomedical
Study
[ 0.99951171875, 0.00018668174743652344, 0.00016963481903076172 ]
[ 0.99853515625, 0.00024127960205078125, 0.001094818115234375, 0.00006753206253051758 ]
Myocardial contraction bands can be observed in multiple circumstances of death, as a result of artefactual causes such as poor sampling techniques (e.g., a low-temperature fixator). In experimental studies on ligated pig hearts, bands were found with abundance in the first 20–30 min . In the study conducted by Morita et al. on necropsy fragments of human heart, in order to differentiate pathological contraction bands from artefactual ones by immunohistochemical analysis, CCC9-positive reactions (complement C9 fraction) and SIRT 1-negative reactions were found in cases of myocarditis and myocardial ischemia, and in the rest of the cases, who benefited from cardiopulmonary resuscitation but had various causes of death, a positive reaction was found only in the case of SIRT 1, concluding that the positive CCC9 marker allows for the differentiation between myocardial changes in acute myocardial infarction and myocardial changes occurring after attempts at cardiopulmonary resuscitation or other terminal conditions .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277133_p27
PMC11277133
sec[2]/sec[0]/p[1]
3.1. Agonal Myocardial Immunohistochemical Changes
2.988281
biomedical
Study
[ 0.998046875, 0.00036263465881347656, 0.00151824951171875 ]
[ 0.9833984375, 0.01473236083984375, 0.0015926361083984375, 0.00028896331787109375 ]
Depending on the immunohistochemical reaction obtained, they proposed the following classification of contraction bands ( Table 3 ).
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277133_p28
PMC11277133
sec[2]/sec[0]/p[2]
3.1. Agonal Myocardial Immunohistochemical Changes
2.646484
biomedical
Other
[ 0.99755859375, 0.0005660057067871094, 0.002044677734375 ]
[ 0.472412109375, 0.51904296875, 0.00629425048828125, 0.00231170654296875 ]
In addition, CCC9 is detectable from the early stage of acute myocardial infarction. Classical hematoxylin–eosin staining does not allow the detection of changes in this regard.
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277133_p29
PMC11277133
sec[2]/sec[0]/p[3]
3.1. Agonal Myocardial Immunohistochemical Changes
4.113281
biomedical
Study
[ 0.99951171875, 0.00019371509552001953, 0.00022208690643310547 ]
[ 0.9970703125, 0.00023853778839111328, 0.00281524658203125, 0.00007206201553344727 ]
Therefore, a myocardial-positive CCC9 immunohistochemical reaction is a reliable element supporting early acute myocardial infarction , while also allowing the differential diagnosis of agonal changes . Sabatosso et al., in their comparative immunohistochemical study of cardiac changes in myocardial ischemia, acute myocardial infarction, and hanging, using necropsy casework, found that myoglobin and TnT showed no expressions, supporting the statistically significant differences between the three groups. This, for example purposes, makes the specificity of immunohistochemical markers in humans inconclusive. Moreover, in the same study, a greater positive expression of Cx34 and Jun B markers was observed in cases of hanging than in cases of acute myocardial infarction or myocardial ischemia, demonstrating their non-specificity in humans, unlike experimental animal studies .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277133_p30
PMC11277133
sec[2]/sec[1]/p[0]
3.2. Myocardial Immunohistochemical Changes Following Autolysis and Putrefaction
3.794922
biomedical
Study
[ 0.9990234375, 0.00023126602172851562, 0.0008625984191894531 ]
[ 0.6171875, 0.37744140625, 0.004558563232421875, 0.0005946159362792969 ]
In general, immunohistochemical results can also be influenced by biological changes during a prolonged postmortem interval with subsequent autolysis and putrefactive phenomena, which, against the background of acidosis, determines the activation of intracellular enzymes with the degradation of protein structures .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999995
PMC11277133_p31
PMC11277133
sec[2]/sec[1]/p[1]
3.2. Myocardial Immunohistochemical Changes Following Autolysis and Putrefaction
4.117188
biomedical
Study
[ 0.99951171875, 0.0002491474151611328, 0.0003230571746826172 ]
[ 0.9775390625, 0.00038552284240722656, 0.0220489501953125, 0.00012993812561035156 ]
In this regard, the study by Thomsen and Held demonstrated that the marker C5b-9 can be detected in the myocardium at a postmortem interval of up to 11 days . Ortmann et al. also obtained a positive reaction for C5b-9 at postmortem intervals of about 8 weeks . Fibronectin and myoglobin could be detected immunohistologically at approximately 3 days postmortem at the myocardial level . However, these studies were not case-controlled and were performed on a small number of subjects, which may make it difficult to differentiate between false-positive results, against the background of protein degradation by autolysis and putrefaction, and positive results in a real pathological context following acute myocardial infarction . Hu et al., in their study in which they analyzed the expression of desmin, actin, and myoglobin on heart fragments, stored at 4C and analyzed at different postmortem intervals, found that the postmortem interval had a significant influence on the experience of the markers studied, with the markers being resistant to autolysis only for an interval of 2 days postmortem .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277133_p32
PMC11277133
sec[2]/sec[2]/p[0]
3.3. Perimortem Catecholamines
4.179688
biomedical
Study
[ 0.99951171875, 0.00021028518676757812, 0.0002378225326538086 ]
[ 0.9912109375, 0.0003268718719482422, 0.008209228515625, 0.00011050701141357422 ]
High levels of catecholamines (either exogenous or endogenous, e.g., in pheochromocytoma or brainstem lesions affecting the nucleus tractus solitari ) have myotoxic action. In experimental animal studies, immunohistochemical myocytic apoptotic phenomena were observed 3–6 h after catecholamine injection, and necrotic phenomena were observed at about 18 h. In the experimental study on mice conducted by Goldspink et al., these phenomena were detected as heterogeneous myocardial, and were more pronounced in the left ventricular subendocardial with the predominance of necrosis phenomena and positive caspase 3 and myosin reactions . Another experimental study on rats by Lu et al. noted that an overdose of norepinephrine (NE) can cause severe cardiopulmonary dysfunction, with cardiac immunohistochemical positivity within the first 6 h of TnT and Cx43 at the biventricular and septal levels .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277133_p33
PMC11277133
sec[2]/sec[3]/p[0]
3.4. Gender and Myocardial Ischemic Preconditions
4.105469
biomedical
Study
[ 0.99951171875, 0.00023686885833740234, 0.00016963481903076172 ]
[ 0.99462890625, 0.0002644062042236328, 0.005138397216796875, 0.0000978708267211914 ]
A person’s gender and pre-existing ischemic conditions impact the severity and course of acute myocardial infarction. Some studies have concluded that the severity of apoptosis and myocardial cell necrosis is more pronounced in men . Even experimental studies in mice have detected smaller areas of acute myocardial infarction in females than in males . Pre-existing ischemic conditions refer to transient myocardial ischemic episodes in the subject’s pathological history and have been found to have a protective role for subsequent cardiac ischemic injury, resulting in infarction in a smaller area and a better preservation of left ventricular function . In the experimental case–control study conducted by Scholl et al. in mice, in which they analyzed the expression of dityrosine, TnT, TnI, and Connexin 43 (Cx43) according to the sex of the subjects and pre-existing myocardial ischemic conditions, it was concluded that only pre-existing ischemic conditions had a significant impact on the expression of troponins and Cx43, with sex having no discriminatory influence on the results obtained .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277133_p34
PMC11277133
sec[2]/sec[4]/p[0]
3.5. Overall Considerations
4.03125
biomedical
Review
[ 0.9853515625, 0.007671356201171875, 0.0069732666015625 ]
[ 0.00505828857421875, 0.0008397102355957031, 0.99365234375, 0.0004067420959472656 ]
Overall, the results of this review find their usefulness in forensic practice, systematizing the main immunohistochemical markers to be analyzed in the case of suspicion of an early acute myocardial infarction. After the first 6 h, these markers can be detectable for certain time intervals (see Table 1 —Included studies) with the coexistence of the usual morphological changes in acute myocardial infarction, detectable by classical optical microscopy, which can help better characterize acute myocardial infarction. Many markers show significant changes dependent on the time interval, as has been extensively evaluated elsewhere in extensive, comparative analyses. The studies existing so far in the analyzed databases are mainly experimental and performed on non-human subjects, but have an important contribution to forensic practice in the instance of the suspicion of early acute myocardial infarction, undetected by classical microscopy techniques. In future, the simultaneous analysis of a panel of markers will be necessary to increase diagnostic accuracy, requiring appropriate laboratory infrastructure and reagents . Also, extensive comparative studies are needed to analyze the specificity of the presented immunohistochemical markers.
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999999
PMC11277133_p35
PMC11277133
sec[2]/sec[4]/p[1]
3.5. Overall Considerations
4.070313
biomedical
Review
[ 0.99169921875, 0.005359649658203125, 0.003047943115234375 ]
[ 0.005847930908203125, 0.0016031265258789062, 0.9921875, 0.0006036758422851562 ]
The limitations of this review are represented by the absence of the specificity of these markers for early acute myocardial infarction, being rather markers of acute myocyte necrosis in general, regardless of etiology, and the small number of studies aimed at the immunohistochemical analysis of acute myocardial infarction more recent than 6 h. Another limit is represented by interspecies variability in marker analysis, with most markers being analyzed on non-human subjects. In experimental studies, human tissue fragments may show limited specificity. Consequently, this requires the survey of a larger number of tissue samples, the simultaneous analysis of several immunohistochemical markers, and the careful analysis of survey data regarding perimortem events, resuscitation, and postmortem intervals.
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277133_p36
PMC11277133
sec[3]/p[0]
4. Materials and Methods
4.074219
biomedical
Review
[ 0.99853515625, 0.0009765625, 0.0006380081176757812 ]
[ 0.38623046875, 0.002094268798828125, 0.61083984375, 0.0007195472717285156 ]
We conducted a study in adherence to the PRISMA guidelines for reporting systematic literature reviews. In this regard, we searched the Web of Science and PubMed databases from inception to 2023, using the keywords “myocardial infarction” and “immunohistochemistry”. The inclusion criteria were observational studies that analyzed the effectiveness of immunohistochemical markers in the postmortem diagnosis of early acute myocardial infarction (the first 6 h after the occurrence) and exposed the laboratory techniques and materials used. We excluded reviews and meta-analyses. We also analyzed the reference lists of the studies included from the 2 databases to add potential supplementary studies to this literature review. The main analyzed elements in each study were as follows: the number of subjects, type of subjects (humans/non-humans), age of the subjects, documented pathologies of the subjects, symptom–death time interval, laboratory equipment and laboratory techniques used (see Supplementary Table S1 —Laboratory methodology in the included studies), and immunohistochemical marker(s) analyzed. The review is registered in the Open Science Framework: https://doi.org/10.17605/OSF.IO/2GWUT .
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277133_p37
PMC11277133
sec[4]/p[0]
5. Conclusions
4.011719
biomedical
Study
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[ 0.96533203125, 0.006542205810546875, 0.0278778076171875, 0.00033545494079589844 ]
Sudden cardiac death still poses a challenge to necropsy practice. Although the pathophysiological mechanisms of acute myocardial infarction have been carefully analyzed and unraveled through studies conducted on both human and animal subjects, we have a reduced range of feasible immunohistochemical markers useful in diagnosing early acute myocardial infarction in human subjects. Extensive studies on human necrotic tissue fragments are required with the simultaneous analysis of several immunohistochemical markers, including survey data for perimortem events, resuscitation, and postmortem intervals in the context of a uniform laboratory methodology.
[ "Oana-Maria Isailă", "Oana Mihaela Ion", "Robert Luta", "Raluca Catinas", "Ana Ionita", "Diana Haisan", "Sorin Hostiuc" ]
https://doi.org/10.3390/ijms25147625
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277145_p0
PMC11277145
sec[0]/p[0]
1. Introduction
3.470703
biomedical
Other
[ 0.998046875, 0.0003867149353027344, 0.0017747879028320312 ]
[ 0.13671875, 0.8134765625, 0.048828125, 0.0011196136474609375 ]
As skin ages, it undergoes numerous changes, with facial wrinkles being one of the most prominent effects . The causes of facial wrinkles include a combination of factors such as UV exposure, oxidative damage, and heat, leading to the degradation of matrix proteins such as elastin fibers and a decrease in moisture content . Additionally, wrinkles are formed and become more pronounced over time due to the continuous contraction of facial muscles .
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p1
PMC11277145
sec[0]/p[1]
1. Introduction
4.253906
biomedical
Study
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Muscle contraction is induced by acetylcholine (ACh), a neurotransmitter released from nerve cells . ACh is released into the space between muscles and nerves when synaptic vesicles fuse with the cell membrane through the action of the SNARE (Soluble N-ethylmaleimide-sensitive factor attachment protein receptor) complex within nerve cells. Released ACh binds to nAChR or muscarinic acetylcholine receptor (mAChR) on the muscle cell surface to induce muscle contraction . Facial wrinkles are formed by muscle contraction through the binding of muscle nAChRs (subtypes α1, β1, δ, γ, and ε) present on the skeletal muscle cell surface . These facial wrinkles can be effectively improved through muscle relaxation by inhibiting the formation of SNARE complexes to inhibit ACh release, or by binding to nAChRs instead of ACh to inhibit the action of ACh. Utilizing these methods, cosmetic peptides such as Argireline ® (Lipotec, Spain) and SYN ® -AKE (DSM, Netherlands) are currently widely used for wrinkle improvement . According to previous research results, Argireline ® is a peptide that binds to the SNARE complex and inhibits ACh secretion, and the wrinkle reduction effect is known to be about 30% . SYN ® -AKE, which binds to nAChRs and inhibits the action of ACh, has a reported IC 50 of 180 μM , and both ingredients have been proven to be effective in improving wrinkles.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277145_p2
PMC11277145
sec[0]/p[2]
1. Introduction
4.164063
biomedical
Study
[ 0.99951171875, 0.00036454200744628906, 0.000186920166015625 ]
[ 0.9990234375, 0.00021851062774658203, 0.0005278587341308594, 0.00008559226989746094 ]
Therefore, this study aimed to develop a more efficient novel peptide that produces a potential anti-aging effect while reducing facial skin wrinkles by inhibiting muscle contraction. We screened for peptides specific to the nAChR subunit α1 using a peptide phage display technique. After several rounds of enrichment, ELISA was performed to confirm the sequence of the selected peptides, and then their binding affinity to the nAChR subunit α1 was measured using surface plasmon resonance (SPR). Through a process of verifying the inhibition of the nicotine response in a concentration-dependent manner using cells expressing nAChRs, it was confirmed whether the peptides could bind to nAChRs and inhibit the action of ACh. We also evaluated the impact of these peptides on collagen protein synthesis affecting skin elasticity and examined the expression of factors influencing skin hydration, HAS2, and AQP3. Finally, the safety of the peptides was confirmed by evaluating skin irritability and eye irritability, and the anti-aging effects of the peptides were verified through clinical efficacy tests on facial wrinkles.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p3
PMC11277145
sec[1]/sec[0]/p[0]
2.1. The Screening of Phage Peptides That Specifically Bind to the Surface of nAChR Subunit α1
4.09375
biomedical
Study
[ 0.99951171875, 0.00028514862060546875, 0.00019943714141845703 ]
[ 0.99951171875, 0.00017774105072021484, 0.0003027915954589844, 0.00006693601608276367 ]
In this study, we first screened and identified peptides that could specifically bind to the surface of nAChR subunit α1 using the 6-mer phage display peptide library. The output/input phage ratio was calculated for each round of nAChR subunit α1 biopanning. The ratio was decreased until Round 3 and then increased during the fourth round . The data showed that the number of fourth-round phages was 3.94 times higher than that of the third-round experiment ( Table 1 ). The 310 phage clones from the fourth round of biopanning underwent ELISA to select the nAChR subunit α1 binding phage clones. As shown in Figure 1 b, nine phage clones had a very high binding affinity to the nAChR subunit α1 compared to that of streptavidin. Thus, these nine positive phage clones were propagated, and their DNA was sequenced to confirm the amino acid sequences of the peptides ( Table 2 and Table S1 ).
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277145_p4
PMC11277145
sec[1]/sec[1]/p[0]
2.2. The Binding Affinity of the Phage Display-Derived Peptides with the nAChR Subunit α1
4.148438
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Study
[ 0.99951171875, 0.0002830028533935547, 0.0001798868179321289 ]
[ 0.99951171875, 0.000225067138671875, 0.0003070831298828125, 0.00007992982864379883 ]
We performed an SPR analysis to confirm the binding affinities of the nine peptides to nAChR subunit α1. The nAChR subunit α1 was immobilized on a sensor chip, and we then tested the nine peptides with the same concentration (5 μM) using SPR analysis and found that peptide 289 had the highest amount of resonance units (RU) compared to the other peptides . These data show that peptide 289 has a stronger binding affinity to nAChR subunit α1 than the other peptides. Based on these results, we selected peptide 289 as a novel peptide (named Medipep). To measure the binding affinity of peptide 289 to nAChR subunit α1, SPR analysis was performed at different concentrations . According to the results of kinetic analysis, peptide 289 showed an association rate constant (ka) of 1.49 × 10 4 M −1 s −1 and a dissociation rate constant (kd) of 9.06 × 10 −3 s −1 , yielding an equilibrium dissociation constant (KD) of 6.09 × 10 −7 M ( Table 3 ).
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277145_p5
PMC11277145
sec[1]/sec[2]/p[0]
2.3. The Effect of a Novel Peptide on Inhibiting the Action of ACh by nAChR Interaction
4.152344
biomedical
Study
[ 0.99951171875, 0.00022733211517333984, 0.00019252300262451172 ]
[ 0.99951171875, 0.00021266937255859375, 0.0003674030303955078, 0.00006347894668579102 ]
We evaluated whether the peptide could bind to muscle nAChR and inhibit the action of ACh. For the evaluation, we used nicotine, which binds to the same nAChR site as ACh, instead of ACh, which binds to both mAChRs and nAChRs. Nicotine acts as an agonist in the same way as ACh . As shown in Figure 3 , our peptides showed nicotine responses of 97%, 42.5%, and 3% at concentrations of 1, 2, and 5 μM, respectively. In addition, the positive control, SYN ® -AKE, showed nicotine responses of 98%, 52.5%, and 3.5% at concentrations of 100, 250, and 500 μM. That is, our peptide showed a similar nicotine inhibitory effect even at about 1/100 of the positive control’s concentration, and we confirmed that the nicotine response was inhibited in a concentration-dependent manner. Since nicotine acts as an agonist in the same way as ACh, these results suggest that the peptide can inhibit the action of ACh by binding to nAChRs.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
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0.999999
PMC11277145_p6
PMC11277145
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2.4. The Effects on Collagen Production and AQP3/HAS2 Gene Expression by a Novel Peptide
4.097656
biomedical
Study
[ 0.99951171875, 0.0002472400665283203, 0.00025010108947753906 ]
[ 0.99951171875, 0.0001513957977294922, 0.00024247169494628906, 0.000054717063903808594 ]
To evaluate the effects of our novel peptide (Medipep) on collagen expression, enzyme-linked immunosorbent assay (ELISA) analysis was performed on dermal fibroblasts. As a result, collagen expression slightly increased at 1000 ppm, and the extent was statistically significant . In addition, to confirm the expressions of the aquaporin-3 (AQP3) and hyaluronan synthase-2 (HAS2) genes in the treated keratinocytes, in vitro real-time PCR was performed. As shown in Figure 4 b,c, significant increases in AQP3 and HAS2 gene expression were observed. Therefore, we confirmed that the peptide affects increased collagen production as well as AQP3 and HAS2 gene expression.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999999
PMC11277145_p7
PMC11277145
sec[1]/sec[4]/p[0]
2.5. Safety Evaluation of Novel Peptide: Cytotoxicity, Skin Irritation, and Eye Irritation
4.183594
biomedical
Study
[ 0.99853515625, 0.001064300537109375, 0.00020682811737060547 ]
[ 0.99853515625, 0.0009412765502929688, 0.00036644935607910156, 0.0001474618911743164 ]
Prior to carrying out the clinical trial, cytotoxicity, skin irritation, and eye irritation were evaluated to confirm the safety of the peptide. During the experiment, the peptide showed no toxicity at a concentration of 0.1 ppm to 100 ppm . Skin reactivity, derived from human patch testing in 31 subjects, is summarized in Table S2 . The results indicated a skin reactivity score of 0.0 ± 0.00 for a novel peptide concentration ranging from 0.1 ppm to 100 ppm, signifying non-irritation in accordance with the criteria outlined in the Frosch and Kligman and COLIPA guidelines. In addition, the eye irritation potential of the peptide was evaluated using the MCTT HCE TM evaluation method using a human cornea model, and it was found to be non-irritating in the concentration range of 0.1 ppm to 100 ppm ( Table S3 ).
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277145_p8
PMC11277145
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2.6. Anti-Wrinkle Efficacy of Novel Peptide in Clinical Study
4.136719
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Study
[ 0.98193359375, 0.0174560546875, 0.0004565715789794922 ]
[ 0.97998046875, 0.017059326171875, 0.0009946823120117188, 0.0018520355224609375 ]
To further evaluate the anti-aging efficacy of the peptide, a clinical study was performed on normal, healthy volunteers. A total of 20 female volunteers with a mean age (±standard deviation (SD)) of 49.4 (±5.08) (min. 40; max. 65) completed the study, wherein the elasticity of each subject’s cheek area, density of the dermis, and crow’s feet wrinkles were measured. After three weeks and six weeks of usage, measured elasticity and dermis density were significantly improved compared to the baseline value .
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p9
PMC11277145
sec[1]/sec[5]/p[1]
2.6. Anti-Wrinkle Efficacy of Novel Peptide in Clinical Study
4.09375
biomedical
Study
[ 0.998046875, 0.0016937255859375, 0.00036406517028808594 ]
[ 0.9990234375, 0.0005168914794921875, 0.00048089027404785156, 0.00013124942779541016 ]
The evaluation revealed notable improvements in various parameters associated with crow’s feet wrinkles. Specifically, the overall size, depth, and maximum depth of crow’s feet wrinkles exhibited statistically significant reductions at both the 3rd and 6th weeks post-product application compared to the baseline measurements . In detail, there was a significant decrease in overall size of 31.5%, a reduction in depth of 30.1%, and a noteworthy decline in maximum depth of 27.9% ( Table 4 ).
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
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en
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PMC11277145_p10
PMC11277145
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3. Discussion
3.986328
biomedical
Review
[ 0.9990234375, 0.0003323554992675781, 0.0008373260498046875 ]
[ 0.3369140625, 0.09326171875, 0.56884765625, 0.0009527206420898438 ]
As the skin ages, it undergoes gradual morphological and physiological declines, exhibiting clear signs of aging . Skin aging can be classified into two types: intrinsic aging, a natural phenomenon dictated by genetics as we age, and extrinsic aging, which occurs due to external environmental factors, predominantly caused by UV exposure . The combination of intrinsic and extrinsic factors leads to the characteristic indicator of aged skin: wrinkles . Wrinkles arise from a variety of causes, including damage to matrix proteins such as elastin fibers; dehydration; muscle movement in the face; UV exposure; oxidative damage; and heat .
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
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en
0.999997
PMC11277145_p11
PMC11277145
sec[2]/p[1]
3. Discussion
4.0625
biomedical
Study
[ 0.99951171875, 0.0002237558364868164, 0.00028896331787109375 ]
[ 0.99951171875, 0.0003046989440917969, 0.00022327899932861328, 0.000058710575103759766 ]
This study focused primarily on the movement of facial muscles as a cause of wrinkle formation. Facial muscle movement, induced by the neurotransmitter ACh released from nerve cells, involves the fusion of synaptic vesicles mediated by the SNARE complex and the subsequent release of ACh into the synapse between muscle and nerve . The released AChs bind to nAChRs on muscle cells, causing contraction . Repeated muscle contraction over time leads to the formation and deepening of wrinkles . In pursuit of potential anti-aging effects such as reducing facial skin wrinkles, this study aimed to develop a novel peptide that inhibits muscle contractions.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p12
PMC11277145
sec[2]/p[2]
3. Discussion
4.136719
biomedical
Study
[ 0.99951171875, 0.00017917156219482422, 0.00016307830810546875 ]
[ 0.9990234375, 0.0003292560577392578, 0.0005054473876953125, 0.00006312131881713867 ]
Phage display technology is widely used and is a useful method for finding new peptides . This technique involves inserting a peptide coding gene into a phage gene to create a library of billions of peptides, allowing for the simultaneous screening of up to a billion different peptides against a desired target . We set the target binding site to the nAChR subunit α1 and performed the phage display. Clones were collected through a total of four enrichment processes, and each clone was measured by ELISA, after which nine clones were selected in order of high specific binding affinity to the nAChR subunit α1. SPR analysis was used to measure the binding force between the nine peptides and the nAChR subunit α1. SPR analysis is a general method used to measure the binding force between a peptide and a receptor using association and dissociation rates . The best peptide (peptide 289, Medipep) was selected by measuring the SPR of each of the nine peptides, and the binding affinity of the best peptide to the nAChR subunit α1 was 609 nM.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277145_p13
PMC11277145
sec[2]/p[3]
3. Discussion
4.171875
biomedical
Study
[ 0.99951171875, 0.00018334388732910156, 0.00017702579498291016 ]
[ 0.99951171875, 0.0002677440643310547, 0.000324249267578125, 0.000060677528381347656 ]
In previous studies, ACh has been shown to bind to the C-loop of subunit α1, the allosteric site of the nAChR . Based on this, we treated TE671 cells expressing nAChRs with our peptide and nicotine (which acts as an agonist in the same way as ACh), and it was confirmed that the nicotine response was inhibited in a concentration-dependent manner . The results showed that while the SYN ® -AKE peptide required a very high concentration of 500 µM to inhibit the nicotinic response, our peptide demonstrated similar inhibition at a concentration approximately 1/100 of that. This allows us to hypothesize that our peptide binds to the C-loop of subunit α1, the allosteric site of nAChR, thereby inhibiting ACh from binding to the receptor and attenuating wrinkle formation by muscle contraction. However, to prove this hypothesis, we believe that it will be necessary to identify the exact mechanism through three-dimensional structural modeling during further study.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
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en
0.999998
PMC11277145_p14
PMC11277145
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3. Discussion
4.25
biomedical
Study
[ 0.99951171875, 0.00035071372985839844, 0.00036597251892089844 ]
[ 0.84423828125, 0.0916748046875, 0.06280517578125, 0.0010528564453125 ]
The formation of wrinkles due to facial muscle movement is more pronounced in skin that is dry and less elastic, highlighting the importance of maintaining skin elasticity and hydration . Collagen, along with elastin, forms the structural support that maintains the strength and elasticity of the skin and is one of the basic components of the extracellular matrix (ECM) in the skin . Collagen loss leads to degeneration of the skin’s support structure, reducing its elasticity . Additionally, hyaluronic acid, synthesized by the enzyme HAS2, fills the space between collagen and elastin fibers in the dermis, storing moisture to hydrate and protect the skin . Aquaporins—water channel proteins on the cell membrane—primarily regulate the movement of water between the inside and outside of cells, with AQP3 mainly present in the skin, maintaining hydration .
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
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https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277145_p15
PMC11277145
sec[2]/p[5]
3. Discussion
4.171875
biomedical
Study
[ 0.9990234375, 0.0007791519165039062, 0.00015223026275634766 ]
[ 0.99853515625, 0.0006723403930664062, 0.00048089027404785156, 0.00015795230865478516 ]
Therefore, when we evaluated the peptide’s effects on collagen protein synthesis and on HAS2 and AQP3 gene expression in the skin, we found that collagen synthesis and gene expression were significantly increased. Additionally, in vitro fibroblast testing confirmed that the peptide was non-cytotoxic up to concentrations of 100 μg/mL. Additionally, the safety of the peptide was confirmed through eye and skin irritation tests. Based on these results, a human clinical trial showed significant improvement in wrinkles after 3 weeks of use and a noticeable reduction in skin wrinkles after 6 weeks, with a decrease in overall size of 31.5%, a reduction in depth of 30.1%, and a noteworthy decline in maximum depth of 27.9%.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
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en
0.999996
PMC11277145_p16
PMC11277145
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3. Discussion
3.847656
biomedical
Study
[ 0.99951171875, 0.0002834796905517578, 0.0002396106719970703 ]
[ 0.998046875, 0.0011777877807617188, 0.000492095947265625, 0.00010472536087036133 ]
In conclusion, the results of this study demonstrate that the peptide (named Medipep) produces a wrinkle-improving effect by controlling factors related to muscle contraction, skin elasticity, and moisture, confirming its anti-aging effect in clinical trials.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
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https://creativecommons.org/licenses/by/4.0/
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0.999998
PMC11277145_p17
PMC11277145
sec[3]/sec[0]/p[0]
4.1. Materials and Reagents
3.113281
biomedical
Study
[ 0.99853515625, 0.00028824806213378906, 0.0010242462158203125 ]
[ 0.9384765625, 0.06011962890625, 0.0009641647338867188, 0.0005292892456054688 ]
The ligand used recombinant human acetylcholine receptor subunit α1, Cat. No. MBS963713 (MyBioSource, San Diego, CA, USA), and SYN ® -AKE used dipeptide diaminobutyroyl benzylamide (Cayman Chemical, Ann Arbor, MI, USA). All peptides used in this study were synthesized and supplied by Anygen (Gwangju, Republic of Korea). Phosphate-buffered saline (PBS), Tween 20, polyethylene glycol (weight-averaged molecular weight (MW) = 8000), LB agar, and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). All reagents and cell culture media were purchased from WELGENE (Gyeongsan-si, Republic of Korea). The INCI name for the novel peptide is Palmitoyl Hexapeptide-93. It was synthesized by Incospharm (Daejeon, Republic of Korea), and clinical trials were conducted with a content of 500 ppm (w/v) .
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277145_p18
PMC11277145
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4.2. Cell Culture
4.074219
biomedical
Study
[ 0.99951171875, 0.00016987323760986328, 0.00027108192443847656 ]
[ 0.98193359375, 0.01702880859375, 0.0009403228759765625, 0.0002263784408569336 ]
TE671 rhabdomyosarcoma cell line was purchased from Korea Cell Line Bank (Seoul, Republic of Korea). Dermal fibroblasts (Hs68) were purchased from American Type Culture Collection. Cells were cultured in Dulbecco’s Modified Eagle’s medium (DMEM) and medium containing 10% fetal bovine serum (FBS), 1% penicillin-streptomycin at 37 °C, and up to 5% CO 2 . Human skin keratinocytes (HEKn) were purchased from Thermo Fisher Scientific (Waltham, MA, USA) and cultured using keratinocyte growth medium, keratinocyte-SFM supplemented with epidermal growth factor (EGF) and bovine pituitary extract (BPE) at 37 °C, and up to 5% CO 2 .
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
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https://creativecommons.org/licenses/by/4.0/
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0.999996
PMC11277145_p19
PMC11277145
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4.3. Phage Library and Biopanning
4.226563
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Study
[ 0.99951171875, 0.0004165172576904297, 0.00019860267639160156 ]
[ 0.9990234375, 0.00046706199645996094, 0.0003311634063720703, 0.00010627508163452148 ]
The biopanning technique was designed in a previous study and used with some modifications . It is shown in Figure 7 as an affinity selection technology used to identify peptides that bind to a given target. The 6-mer phage display peptide library was constructed through NNK codon-based randomization (N = A or C or G or T; K = G or T). The gene fragments were double digested with SfiI/NotI and cloned into pIGT2 phagemid vectors. The resulting constructs were transformed into Escherichia coli ( E. coli ) cells and the naive library of 6-mer peptides was composed of 1.23 × 10 7 independent 6-mer peptide clones. The 6-mer peptide phage library was prepared using the M13KO7 helper phage. The nAChR subunit α1 was used as the target protein during phage selection. The nAChR subunit α1 was immobilized directly on 96-well high-binding plates overnight at 4 °C. After that, the plates were incubated in blocking buffer (PBS containing 2% BSA) for 2 h at room temperature. The prepared 6-mer peptide phage was added to the plates for 1 h at 30 °C. Unbound phages were removed by washing with PBS containing 0.05% Tween 20 (three times in rounds 1 and 2, four times in rounds 3 and 4). Bound phages were subsequently eluted by incubation with 0.2 M glycine HCl (pH 2.2) for 10 min, followed by immediate neutralization with 1 M Tris HCl (pH 9.0). To perform the next round of biopanning, the eluted phages were used to infect E. coli for 1 h at 37 °C, and helper phages were then added and incubated for 18 h at 37 °C. The culture was then centrifuged at 12,000× g for 10 min at 4 °C. After that, the supernatant was transferred into a new tube with the addition of one-sixth volume of 20% polyethylene glycol (PEG)/2.5 M NaCl and incubated for 1 h at 4 °C. The PEG-precipitated phages were then centrifuged at 12,000× g for 1 h at 4 °C. The phage pellet was resuspended in 1000 μL of PBS, and that was the first nAChR subunit α1-specific sublibrary. At each step, the input and output phage PFU in all the rounds of biopanning were measured.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p20
PMC11277145
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4.4. Enzyme-Linked Immunosorbent Assay (ELISA)
4.128906
biomedical
Study
[ 0.99951171875, 0.00028967857360839844, 0.0001837015151977539 ]
[ 0.9990234375, 0.0004162788391113281, 0.0002529621124267578, 0.00007492303848266602 ]
To identify phage clones, we performed an ELISA assay. After four rounds of in vitro biopanning, 310 were randomly chosen from the titration plate. For this purpose, 5 μg/mL of nAChR subunit α1 was used to coat a Nunc MaxiSorp 96-well plate and left at 4 °C overnight. After blocking with 2% BSA, the plate was washed three times with PBS-T (PBS with 0.05% v / v Tween 20), and the supernatants from the phage culture were applied to the plates and incubated for 1 h at 37 °C. After washing three times, the HRP-conjugated anti-M13 antibody was incubated on the plate for 1 h. The plate was washed three times to get rid of the nonspecific antibody, followed by color development with the addition of the 3,3′,5,5′-tetramethylbenzidine (TMB) substrate. The incubation was stopped by adding 2 M H 2 SO 4 . Finally, the absorbance was measured at 450 nm using a microplate reader. The positive clones were subjected to DNA sequencing using a phagemid primer (5′-GATTACGCCAAGCTTTGGAGC-3′; Bioneer, Daejeon, Republic of Korea).
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p21
PMC11277145
sec[3]/sec[4]/p[0]
4.5. Binding Affinity Using Surface Plasmon Resonance (SPR) Assay
4.164063
biomedical
Study
[ 0.99951171875, 0.00024390220642089844, 0.00016379356384277344 ]
[ 0.9990234375, 0.00041794776916503906, 0.000362396240234375, 0.00007420778274536133 ]
The SPR analysis was performed using a BIACORE 3000 (GE healthcare, Chicago, IL, USA). The nAChR subunit α1 was immobilized on a CM5 sensor chip by injecting the protein diluted in PBS (0.2 M phosphate buffer, 0.027 M KCl, 1.37 M NaCl, pH 7.4) at a concentration of 0.35 mg/mL at a flow rate of 5 μL min −1 . After, 5 μM peptide was injected into running buffer (20 mM Tris, pH 7) and its interaction with nAChR subunit α1 was measured at a flow rate of 30 μL min −1 . For kinetic experiments, peptides diluted in running buffer at concentrations ranging from 100 μL to 200, 400, 600, 1000, and 1200 nM were injected at a flow rate of 30 μL min −1 over the immobilized nAChR subunit α1. The association and dissociation kinetic rate constants (ka and kd) and the equilibrium association constant, KD, were calculated using BIAevaluation 3.1 software .
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p22
PMC11277145
sec[3]/sec[5]/p[0]
4.6. In Vitro Intracellular Nicotine Response Assay
4.15625
biomedical
Study
[ 0.99951171875, 0.0005254745483398438, 0.00018334388732910156 ]
[ 0.99658203125, 0.002979278564453125, 0.0004668235778808594, 0.00017070770263671875 ]
TE671 cells were cultured in DMEM and medium containing 10% FBS, 1% penicillin-streptomycin at 37 °C, and up to 5% CO 2 . An 18 mm coverslip was placed on a 12-well cell culture plate, cultured cells were removed with trypsin, and 1 mL was dispensed into each well to make 2 × 10 4 cells/well, followed by culturing for 4 days. The coverslip with the cultured cells was transferred to a new 12-well plate, mixed with 997 μL of HBSS buffer and 3 μL of Fura-2-AM (Invitrogen, Waltham, MA, USA), and incubated for 15 min. After incubation, the remaining Fura-2-AM was removed by washing three times with HBSS buffer, and 1 mL of HBSS buffer was additionally dispensed. After mounting the coverslip in the chamber (Live Cell Instrument, Namyangju-si, Republic of Korea), 500 μL of HBSS buffer was dispensed, and the peptide and nicotine samples to be tested were adjusted according to the concentration. Then, 10 to 20 cells were selected and captured alternately at 340 nm and 380 nm wavelengths at 500 ms intervals through a fluorescence microscope . The fluorescence ratio at 340/380 nm was calculated using the LAS X program .
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p23
PMC11277145
sec[3]/sec[6]/p[0]
4.7. In Vitro Cell Viability Assay
4.144531
biomedical
Study
[ 0.99951171875, 0.0003561973571777344, 0.0001493692398071289 ]
[ 0.9990234375, 0.0003910064697265625, 0.00045180320739746094, 0.00009125471115112305 ]
The in vitro cytotoxicity of the novel peptide on cells was assessed using the MTT [(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cell viability assay. Briefly, dermal fibroblasts were cultured in 24-well plates at a density of 1 × 10 5 cells/well for 18 h. After the medium was removed, the sample diluted in FBS-free medium was added and cultured for 24 h. Then, after the medium was removed, MTT solution was added at a concentration of 1 μg/mL and left to react for 3 h. Unreacted MTT was removed, 100 μL of DMSO was added to dissolve the formed reaction product, and the absorbance at 540 nm was measured and evaluated using an ELISA reader.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p24
PMC11277145
sec[3]/sec[7]/p[0]
4.8. Measurement of Collagen Production and Gene Expression
4.097656
biomedical
Study
[ 0.99951171875, 0.00022780895233154297, 0.0001666545867919922 ]
[ 0.99853515625, 0.0011768341064453125, 0.00031638145446777344, 0.00008237361907958984 ]
Dermal fibroblasts were cultured in 48-well plates at a density of 2~5 × 10 4 cells/well for 24 h. The existing medium was removed, and the medium containing the sample was added. The culture was incubated for 24 h, after which the cell culture medium was collected and subjected to ELISA assay. The Human Pro-Collagen α1 DuoSet ® ELISA kit used in this test was from R&D Systems (Minneapolis, MN, USA) and was used according to the experimental protocol provided by the manufacturer.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277145_p25
PMC11277145
sec[3]/sec[7]/p[1]
4.8. Measurement of Collagen Production and Gene Expression
4.152344
biomedical
Study
[ 0.99951171875, 0.00024199485778808594, 0.0001742839813232422 ]
[ 0.9990234375, 0.0004940032958984375, 0.0003204345703125, 0.00007134675979614258 ]
For sample processing, keratinocytes were seeded in a 60 mm culture plate at a concentration of 2~5 × 10 5 cells/mL and cultured for 24 h. The growth medium was removed, and the samples were added at different concentrations to keratinocyte-SFM without EGF and BPE, after which the keratinocytes were treated for 24 h. Total RNA was isolated from cultured epidermal keratinocytes using an Easy-blue RNA Extraction Kit, Cat. No. 17061 (iNtRON Biotechnology, Seongnam-si, Republic of Korea). All primers for the quantitative reverse transcription PCR (qRT-PCR) ( Table S4 ) were designed using Primer Express v.1.5 (Applied Biosystems, Waltham, MA, USA) software. Total RNA (2 μg) was reverse transcribed to complementary DNA with an M-MLV reverse transcriptase. The resulting complementary DNA was amplified using a SYBR Green Master Mix kit, Cat. No.EBT-1802 (ELPIS Biotech, Daejeon, Republic of Korea). Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was performed using an AB StepOne Real-Time PCR system (Applied Biosystems, USA). All samples were analyzed using the comparative C t method (2 −ΔΔCt ) for the relative quantification of gene expression and normalization with respect to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p26
PMC11277145
sec[3]/sec[8]/p[0]
4.9. Skin Irritation Test
4.023438
biomedical
Study
[ 0.9990234375, 0.0006470680236816406, 0.00035881996154785156 ]
[ 0.99853515625, 0.00110626220703125, 0.00020360946655273438, 0.00010979175567626953 ]
The skin irritation test was conducted at CRA Korea Co., Ltd. in the Republic of Korea, with the approval serial number CRA21-HIPTRP1001. The patch used was the IQ Ultra Chamber (Chemotechnique Diagnostics, Vellinge, Sweden). The human skin primary irritation test was completed, with all 31 subjects participating until the end of the study. The test area for the product was the back (excluding the spinal area). The patch was applied for 24 h and then removed. The first evaluation was conducted 30 min after patch removal while waiting for the transient erythema to disappear due to removal, and the second evaluation was conducted 24 h after patch removal. The irritation criterion used a measurement evaluation method designed by Frosch and Kligman by applying the reading standards of the International Contact Dermatitis Research Group (ICDRG).
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999995
PMC11277145_p27
PMC11277145
sec[3]/sec[9]/p[0]
4.10. Eye Irritation Test
4.140625
biomedical
Study
[ 0.99951171875, 0.00041794776916503906, 0.00018548965454101562 ]
[ 0.9990234375, 0.00043201446533203125, 0.0003337860107421875, 0.00008624792098999023 ]
To evaluate the eye irritation potential of the peptide, an in vitro experimental model recognized by the OECD as an alternative animal evaluation method, the human cornea model (MCTT HCE TM ), was used. The experiment was conducted according to the standard experimental method provided by the manufacturer, Biosolution (Seoul, Republic of Korea). First, before the experiment, the color interference of the test material was checked, and to begin the experiment, MCTT HCE TM was incubated for 22 h at 5% CO 2 and 37 °C. Next, 40 μL of the peptide was applied to the top of the MCTT HCE TM and left to react for 10 min. Afterwards, the peptide was removed using DPBS, and MCTT HCE TM was transferred to a new medium and cultured for 18 h at 5% CO 2 and 37 °C. After the culture was completed, 200 μL/well of WST-1 solution diluted at a ratio of 1:25 was added to a new plate, MCTT HCE TM was transferred thereon, and 100 μL of WST-1 solution was treated inside. Finally, after blocking the light with foil and culturing for 3 h at 5% CO 2 and 37 °C, the WST-1 solution inside and outside the tissue was collected and transferred to a 96-well plate, and the absorbance was measured at 450 nm.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p28
PMC11277145
sec[3]/sec[10]/p[0]
4.11. Clinical Study
4.117188
biomedical
Other
[ 0.51513671875, 0.47900390625, 0.005817413330078125 ]
[ 0.122314453125, 0.84326171875, 0.004154205322265625, 0.030517578125 ]
The open-label clinical trial was conducted on 20 female Asian subjects with facial wrinkles, aged 40 to 65 years old, and the trial was approved by the Institutional Review Board of CRA Korea Co., Ltd. . All studies complied with the World Medical Association’s Declaration of Helsinki concerning biomedical research involving human subjects. Healthy female volunteers without any skin or systemic diseases were initially enrolled in the study. Subjects who had been treated with retinoids or laser therapy within the last six months, or who had participated in another clinical study, were excluded. After hearing the explanation of the purpose and protocol of the study, all the volunteers signed the informed consent form and participated in the study. A total of 21 female volunteers initially enrolled in the study, but during the study, 1 participant was dropped due to failure to attend the scheduled visit, and 20 participants completed the study. During the first visit, subjects were asked to complete study-related medical record questionnaires to confirm the inclusion and exclusion criteria, and each participant provided written, informed consent. Before the instrumental measurements, participants were asked to rest for at least 30 min in a humidity (50 ± 10% RH)- and temperature (22 ± 2 °C)-controlled room. The torsional elasticity and dermal density of the skin in each subject’s cheek area were measured using a Dermal Torque Meter (DTM310, Dia-Stron, Andover, UK) and an ultrasound device, the DUB Skinscanner (tpm, Lueneburg, Germany) . Wrinkles around the eyes were measured using a topographic skin measurement device (Antera 3D CS: Miravex Limited, Dublin, Ireland) . The participants were administered with 619.9 ± 241.19 mg of essence containing the novel peptide twice a day (morning and noon) for 6 weeks. The concentration of the peptide was 500 ppm, determined based on formulation studies at levels that were found to be non-irritating and non-allergenic during preliminary testing. After using the product, they revisited the clinical institution in the 3rd and 6th weeks, where the same measurements as those taken before product use (week 0) were conducted. Subsequently, the results from before and after product usage were compared and evaluated.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277145_p29
PMC11277145
sec[3]/sec[11]/p[0]
4.12. Statistical Analysis
4.046875
biomedical
Study
[ 0.99951171875, 0.00025916099548339844, 0.00030350685119628906 ]
[ 0.9990234375, 0.0004699230194091797, 0.0002219676971435547, 0.000051021575927734375 ]
Since the number of subjects was lower than 30, a normality test was performed using the Shapiro–Wilk test to properly analyze the experimental results. If normality was satisfied, a parametric test was performed, and if normality was not satisfied, statistical analysis was performed using a non-parametric test. The measurements of skin moisture content, wrinkles around the eyes (overall size), and dermal density before and after using the product were compared using the paired t -test, which is a parametric test, and the Wilcoxon signed-rank test, which is a non-parametric test, to obtain the size results of wrinkles around the eyes (depth/maximum depth). The statistical analysis results were checked for significance with a 95% confidence interval, and IBM SPSS Statistics software, version 28.0 was used as the statistical analysis program.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277145_p30
PMC11277145
sec[4]/p[0]
5. Conclusions
3.896484
biomedical
Study
[ 0.99951171875, 0.0002263784408569336, 0.00028967857360839844 ]
[ 0.9990234375, 0.0006279945373535156, 0.0003879070281982422, 0.00008845329284667969 ]
In this study, we developed a novel peptide that simultaneously regulates muscle contraction, skin elasticity, and moisture, and investigated its specificity and anti-aging effect in vitro. The clinical efficacy of the peptide (named Medipep) as an anti-wrinkle cosmetic ingredient was also proven.
[ "Jinho Bang", "Yul-Lye Hwang", "Mi Yoon Kim", "Jae Nam Yun", "Eujin Hyun", "Min Youl Chang", "Dae Hwan Shin", "Sunghyun Kim", "Jeung-Hoon Lee" ]
https://doi.org/10.3390/ijms25147860
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277157_p0
PMC11277157
sec[0]/p[0]
1. Introduction
4.929688
biomedical
Study
[ 0.99853515625, 0.0010938644409179688, 0.000518798828125 ]
[ 0.955078125, 0.00225067138671875, 0.0416259765625, 0.0011720657348632812 ]
Parkinson’s disease (PD) is a neurodegenerative disorder characterized by the progressive loss of dopaminergic neurons, resulting in motor dysfunction and cognitive impairment. Neuronal cell death under oxidative stress and mitochondrial dysfunction are considered major drivers of PD pathologies . DJ-1’s primary role as an essential mitochondrial protein explains much of its role in PD . Mutations in PARK7 (also called DJ-1) were implicated in familial early-onset and sporadic forms of PD . As an oxidative sensor, DJ-1 plays a critical role in maintaining redox homeostasis in all cells by sensing and modulating cellular oxidative responses. DJ-1 is involved in oxidation stress in direct and indirect modes. For example, DJ-1 binds the subunits of mitochondrial complex I. Therefore, upon DJ-1 depletion, mitochondrial function is drastically reduced. Downregulating DJ-1 in neuronal cells led to increased cell death in response to oxidative stress, ER stress, and proteasome inhibition, while overexpression of DJ-1 reduced cell death . Several oxygen-based modifications to the cysteine residues of DJ-1 create a set of modified variants that are partitioned among cellular compartments (mitochondria, cytosol, nucleus, and exosomes) to conduct downstream functions in signaling and transcription regulation . To protect against cell death from various stressors, the modified DJ-1 protein is engaged in a rich protein interaction network that includes often specific signaling pathways and cell types .
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277157_p1
PMC11277157
sec[0]/p[1]
1. Introduction
4.238281
biomedical
Study
[ 0.99951171875, 0.00016796588897705078, 0.0002551078796386719 ]
[ 0.9853515625, 0.00046896934509277344, 0.0140380859375, 0.0001169443130493164 ]
The DJ-1 has garnered considerable interest due to its multifaceted roles in driving numerous processes . DJ-1 is involved in transcriptional regulation in direct and indirect modes. To gain insights into the role of DJ-1 in cells, diverse cell systems were used. This included PD-related cell models from humans and rodents, such as primary dopaminergic neurons , brain slices, and numerous cell lines . Translocation of DJ-1 to the nucleus and activation of transcription were demonstrated not only in neuronal-like cells (e.g., SH-SY5Y, PC12) and numerous primary cell cultures (e.g., astrocytes, microglia, cortical neurons) but also in routinely used cell lines (HEK293, MIN6, COS7, CHO) . In expanding the role of DJ-1 in cell signaling, it has been argued that its location, and the pattern of its post-translational modifications, determines the effect of DJ-1 in different cell systems , as studied by overexpression or knockdown (KD) (e.g., ).
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277157_p2
PMC11277157
sec[0]/p[2]
1. Introduction
4.867188
biomedical
Review
[ 0.99560546875, 0.0021991729736328125, 0.002178192138671875 ]
[ 0.414794921875, 0.0029010772705078125, 0.58056640625, 0.0016431808471679688 ]
Accumulating evidence validates the role of DJ-1 as an oncogene in cancer biology and, more specifically, in tumorigenesis and cancer progression . The expected impact of DJ-1 on cancer is tightly connected to its role in apoptosis, cell proliferation, and metastasis. For example, the role of DJ-1 in suppressing apoptosis in cancer cells allows for the production of the driving oncoproteins, thereby promoting tumor progression. However, the molecular mechanisms underlying DJ-1-mediated modulation of cancer-related pathways remain elusive. In cancer cells, DJ-1 regulates the JNK pathway and consequently impacts autophagy . This was further confirmed in prostate cancer . DJ-1, through the regulation of androgen receptor (AR) signaling, affected Beclin1-involved autophagy responses via the JNK-dependent pathway. DJ-1 overexpression reduces LC3 transformation and autophagosome formation, while DJ-1 knockdown has the opposite effect. JNK phosphorylation and Bcl2 dissociation are affected by DJ-1 levels. Recent studies have highlighted the transforming activity of cancer cells. Overexpression of DJ-1 enhances colorectal cancer cell proliferation through the cyclin-D1/MDM2-p53 signaling pathway and acts to increase cancer cell survival through the PI3K-AKT pathway . The elevated levels of DJ-1 have been detected in breast, lung, and prostate cancers, making it a potential biomarker for diagnosis and prognosis .
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277157_p3
PMC11277157
sec[0]/p[3]
1. Introduction
4.433594
biomedical
Study
[ 0.99951171875, 0.0002880096435546875, 0.00014793872833251953 ]
[ 0.998046875, 0.00039768218994140625, 0.0012235641479492188, 0.00015723705291748047 ]
DJ-1’s function in the brain has shown its involvement in controlling inflammation . Higher levels of inflammation signatures (including TNF, iNOS, NO, IL-6, cyclooxygenase-2, and p38 phosphorylation) were observed in astrocytes and microglia lacking the DJ-1 mouse model after lipopolysaccharide (LPS) exposure . Similarly, brain slices from DJ-1 knockout mice exposed to IFN-γ exhibited increased levels of IL-6 and TNF, as well as STAT1 phosphorylation . Studies that mimicked glial activation by injecting LPS into the substantia nigra of DJ-1 knockout mice resulted in elevated levels of immune-responsive genes including ICAM-1, IFN-γ, IL-1β, IL-16, IL-17, CXCL11, and more . Reducing DJ-1 levels also accelerated IKK and IkBα phosphorylation, leading to enhanced p65 nuclear translocation in both normal and LPS-exposed cells . The presence of an elevated NF-κB promoter function in DJ-1-depleted cells further confirms DJ-1’s inhibitory function in neuroinflammation .
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999995
PMC11277157_p4
PMC11277157
sec[0]/p[4]
1. Introduction
4.53125
biomedical
Study
[ 0.99951171875, 0.00033926963806152344, 0.0002281665802001953 ]
[ 0.9501953125, 0.00444793701171875, 0.0450439453125, 0.0004029273986816406 ]
Upon viral infection, the innate system is activated, leading to an antiviral defense mechanism mediated by interferon (IFNs). The type I IFNs (IFNα, IFNβ) activate IFN receptors (IFNARs) via JAK1, TYK2, STAT1, and STAT2 signaling pathways, whereas type II IFN (IFNγ) activates IFNγ receptors (IFNGRs), leading to STAT1 phosphorylation and gene expression. Upon viral infection (e.g., Respiratory syncytial virus; RSV), reactive oxygen species (ROS) levels increase and, as a result of changes in cell redox, the JAK-STAT pathway is activated, resulting in the expression of antiviral genes . Elevated ROS signifies the pathogenesis of human diseases such as pulmonary fibrosis and Parkinson’s and Alzheimer’s diseases .
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277157_p5
PMC11277157
sec[0]/p[5]
1. Introduction
4.191406
biomedical
Study
[ 0.99951171875, 0.00030112266540527344, 0.00020253658294677734 ]
[ 0.99951171875, 0.00019061565399169922, 0.0003974437713623047, 0.0000730752944946289 ]
In this study, we tested the impact of changing the naïve level of DJ-1 by suppressing its expression in HEK293 cells. We performed RNA-seq global analysis to assess the role of knockdown using siRNA for DJ-1 levels. We show that siRNA causes a stress response that is converted to an antiviral cascade. The lack of DJ-1 in cells causes a failure in redox homeostasis and the coordinated activation of the STAT pathways that connect a transcriptional wave of an interferon-based antiviral response and a direct effect on mitochondria and RNA metabolism. The potential role of targeting DJ-1 in activating the antiviral response in metastatic cancers is discussed.
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277157_p6
PMC11277157
sec[1]/sec[0]/p[0]
2.1. DJ-1 Response to Oxidative Stress by Translocation to Nucleus
4.144531
biomedical
Study
[ 0.99951171875, 0.00015532970428466797, 0.0002930164337158203 ]
[ 0.900390625, 0.0076751708984375, 0.0916748046875, 0.00038695335388183594 ]
PARK7 (DJ-1) was extensively studied for its function as an oxidative stress sensor and in executing the oxidative stress cellular response. This role was associated mostly with neurons and neuronal-like cells that are over-sensitive to oxidation insults . However, the majority of cell lines express DJ-1 at moderate to high levels . The current notion is that the modification of DJ-1 on cysteine residue is a prerequisite for DJ-1 nuclear function, including its role in cancer and metastasis.
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277157_p7
PMC11277157
sec[1]/sec[0]/p[1]
2.1. DJ-1 Response to Oxidative Stress by Translocation to Nucleus
4.238281
biomedical
Study
[ 0.99951171875, 0.00026226043701171875, 0.00021028518676757812 ]
[ 0.99951171875, 0.00021183490753173828, 0.0004191398620605469, 0.00006794929504394531 ]
To this end, we compared the PARK7/DJ-1 gene expression in SH-SY5Y neuroblastoma cells as a model for neuronal-like cells , and in HEK293 cells as a non-cancerous cell line. We observed comparable expression levels of DJ-1 transcripts in both cells . Notably, cell lines exhibit intrinsic capacity to cope with oxidative stress, with neuronal-like cells being more sensitive than other cells’ origins . There are 76 antioxidant activity annotated genes . Figure 1 C shows representative genes, while the expression of DJ-1 is similar in both cell types, many of the genes (e.g., GPX8, GPX4) are highly expressed in HEK293 but display low expression levels in SH-SY5Y. Interestingly, both cell lines expressed the cytoplasmic enzyme SOD1 to different levels, although only SOD2 is expressed in SH-SY5Y cells. SOD2 is located in the mitochondrial matrix and thus impacts mitochondrial apoptosis and the redox function. We anticipate that each cell displays a unique collection of genes to cope with oxidation, rendering a cell-specific capacity for coping with oxidative stress .
[ "Keren Zohar", "Michal Linial" ]
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2.2. Knockdown Resulted in 8-Fold Suppression of the Native PARK7/DJ-1 Transcript
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[ 0.99951171875, 0.00026106834411621094, 0.00021517276763916016 ]
[ 0.99951171875, 0.00018703937530517578, 0.0002703666687011719, 0.000059545040130615234 ]
We transfected HEK293 cells by siRNA as a negative control (using esiRNA-RLUC, see Section 3 ) and the siRNA methodology for PARK7 (see Section 3 ). The cultured cells were collected 24 h following transfection, and total RNA (>200 nt) was prepared for RNA-seq. Figure 2 A shows the results of the RT-PCR at two time points (26 h and 48 h, post-transfection). It is evident that already 26 h post-transfection DJ-1 transcripts are strongly suppressed. Quantitation of the degree of suppression by the specific siRNA relative to the non-specific control siRNA showed an 8-fold reduction, which was constant for 48 h. Note that the RULC control showed no reduction in DJ-1 transcripts, supporting the efficiency of the knockdown (KD) protocol used.
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
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2.2. Knockdown Resulted in 8-Fold Suppression of the Native PARK7/DJ-1 Transcript
4.058594
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[ 0.99951171875, 0.0002199411392211914, 0.0002658367156982422 ]
[ 0.99951171875, 0.00019669532775878906, 0.0001914501190185547, 0.00005245208740234375 ]
Figure 2 B shows the results using dimensional reduction by principal component analysis (PCA). Each experimental group is represented by three biological triplicates. The resulting PCA shows the non-treated cells (N.T.), mock transfection with esiRNA-RULC (depicted RULC), and cells transfected with esiRNA-PARK7 (DJ-1). The two major components (PC1 and PC2) explained together 42% of the variance . The partitioned of the nine samples according to the treatments validates the quality of the results. For the PCA , we have included all 18k identified transcripts from the RNA-seq results ( Supplementary Table S2 ).
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
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2.2. Knockdown Resulted in 8-Fold Suppression of the Native PARK7/DJ-1 Transcript
3.841797
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[ 0.99951171875, 0.00017631053924560547, 0.0004172325134277344 ]
[ 0.9990234375, 0.000682830810546875, 0.0002130270004272461, 0.00006753206253051758 ]
Using the results from RNA-seq with biological triplicates, we calculated the extent of the knockdown of PARK7. Only 12% of the original basal levels were detected 24 h after introducing the siRNAs .
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
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PMC11277157_p11
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2.3. Knockdown of PARK7/DJ-1 Resulted in Coding and Non-Coding Differential Gene Expression
4.136719
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[ 0.99951171875, 0.00028514862060546875, 0.0002884864807128906 ]
[ 0.99951171875, 0.00014138221740722656, 0.0002570152282714844, 0.00005316734313964844 ]
In analyzing all three experimental groups, we observed a large number of differentially expressed genes. The RNA-seq results reported on 18,158 transcripts, 23% of them are non-coding RNAs . Figure 3 summarizes DEG findings for the three experimental pairs. Only statistical threshold genes with FDR < 0.05 are included. There are 527 DEGs between KD DJ-1 and the untreated cells (N.T.), and 316 DEGs between cells of KD DJ-1 and cells introduced with non-specific siRNA RULC. Only 92 DEGs were reported between the siRNA of RULC and N.T. cells. Among these reliable results, transcripts that were differentially expressed by log 2 (FC) > 0.5 (i.e., >|1.412| fold) were labeled as upregulated and downregulated transcripts. The numbers of these transcripts are shown in Figure 3 Only a modest number of DEGs were identified between siRNA RULC and untreated (NT) cells ; in contrast, a substantial number of DEGs were detected for the siRNA of DJ-1 vs. siRNA RULC (316 genes). We concluded that many DEGs are associated with the effect of the KD of DJ-1, and are not attributed to the unavoidable effects of the siRNA protocol perse. The RNA-seq significant results are listed in Supplementary Table S1 .
[ "Keren Zohar", "Michal Linial" ]
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2.4. Non-Specific siRNA Induces Interferon Signaling and the Antiviral Response
4.28125
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[ 0.99951171875, 0.0002868175506591797, 0.00019633769989013672 ]
[ 0.9990234375, 0.000324249267578125, 0.0003333091735839844, 0.00008475780487060547 ]
Upon transfection of the HEK293 with siRNA of RULC, we observed a coordinated induction of genes that are associated with innate immunity and more specifically with interferon response. This observation most likely reflects the non-specific induction of cells to the delivery system for the duplex of RNA molecules and the activation of the dsRNA pathway of viral infection. Testing the induction of RULC vs. naïve, non-treated cells shows the upregulated set of coding transcripts . The network is indicative of the activation of type I Interferons (IFN), which act through the activation of the JAK/STAT signaling cascade to trigger an antiviral response. Note that STAT1 acts as a hub for several transcription factors that belong to the ATF, EGR, and FOS families. The ATF3 is a regulator that links inflammation, oxidative stress, and immune responses .
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
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2.5. Knockdown of PARK7/DJ-1 Strongly Boosted the Antiviral Response
3.65625
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[ 0.99951171875, 0.00018656253814697266, 0.0004858970642089844 ]
[ 0.99755859375, 0.001983642578125, 0.00026226043701171875, 0.00009006261825561523 ]
Figure 5 A shows a Venn diagram of the experiment groups. It shows results from comparing the KD of DJ-1 vs. the background of the siRNA of RULC and the non-treated cell. Among the genes that were changed with esiRNA RULC negative control, 45 of them were also induced by using esiRNA for DJ-1.
[ "Keren Zohar", "Michal Linial" ]
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2.5. Knockdown of PARK7/DJ-1 Strongly Boosted the Antiviral Response
4.21875
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[ 0.99951171875, 0.00034332275390625, 0.00025010108947753906 ]
[ 0.99951171875, 0.00016987323760986328, 0.0003788471221923828, 0.0000699758529663086 ]
Figure 5 B shows an MA plot of the differential expressed transcripts. Highlighted are set genes that meet the threshold for FDR and FC. Inspection of these genes emphasizes the occurrence of viral-like induction that spans all levels of expression ( y -axis). Notably, many of the genes were strongly induced (>10-fold). Among the genes with the strongest induction are the interferon (INF) genes and their modulators. For example, the set of OAS genes that recognize dsRNA as a pathogen-associated molecular pattern (PAMP). The induction of OAS1, OAS2, and OASL, which, as main sensors for viral infections, explains the induction of numerous interferon-induced proteins (e.g., IFIT1, IFIT2, IFI44, MX1, MX2). This coordinated signature argues for further amplification of the antiviral response following the KD of DJ-1. Figure 5 C shows the strong connectivity of the induced genes using STRING analysis. The analysis used a filtered set by removal of the non-specific set and included the rest of the 275 genes that were upregulated as a result of KD by the siRNA of DJ-1 ( Supplementary Table S3 ).
[ "Keren Zohar", "Michal Linial" ]
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2.5. Knockdown of PARK7/DJ-1 Strongly Boosted the Antiviral Response
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[ 0.9990234375, 0.00041961669921875, 0.0003459453582763672 ]
[ 0.9990234375, 0.00048470497131347656, 0.0005664825439453125, 0.00012254714965820312 ]
For partitioning the DJ-1 dependent transcriptomic signature from that of the siRNA response perse (i.e., siRNA RULC), we also show in a unified network the genes identified by the siRNA RULC and the KD DJ-1 set . The sub-networks that are unique to cells of the KD of DJ-1 are highlighted. The DJ-1 induced the protein–protein interaction (PPI) network is composed of several components: (i) The presenting antigen, which includes genes such as TAP1 and TAP2 that together form the TAP protein complex (transporter associated with antigen processing), along with numerous human leukocyte antigen (HLA) genes of the major histocompatibility complexes (MHC). (ii) The histone with H2 and H4 genes that are indicative of epigenetic, gene expression, and chromatin reorganization. (iii) The main component of interferon and the innate immune response. The hub proteins include the STAT3, STAT2, IRF1, and INFB1 genes. Specifically, IRF1 (interferon regulatory factor 1) is a transcription factor and activator of interferons alpha and beta and gamma. (iv) Components of the immunoproteasome (e.g., PSME1, PSMB9) that are induced by γ-INF. The interferon-γ induces apoptotic mechanisms, nitric oxide production, and antigen-presenting pathways (i.e., MHC class I and II with numerous HLA genes). In addition, IRF1 plays a role in apoptosis and tumor suppression. The result of the activation of type I interferon is shown in the network by the increase in inflammatory cytokines and chemokines. Note the activation of smaller connected components such as the BTN (butyrophilin) gene set, which belong to the major histocompatibility complex (MHC)-associated genes, and genes from the classical pathway of the complement system (e.g., C1, C2).
[ "Keren Zohar", "Michal Linial" ]
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2.6. Knockdown of PARK7/DJ-1 Exposed Downregulation of Genes with Membrane-Associated Functions
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[ 0.99951171875, 0.0004138946533203125, 0.0002551078796386719 ]
[ 0.9990234375, 0.0002646446228027344, 0.000560760498046875, 0.00012022256851196289 ]
We identified a small set of genes that was substantially downregulated along with the KD of DJ-1 (57 genes). There is no sign of the involvement of the innate system among the downregulated genes . We note that the degree of downregulation is modest. Inspecting the network shows that on average, the node degree is low, even at a relaxed threshold of PPI confidence (score > 0.4). The gene that was maximally downregulated is CERK (ceramide kinase; 2.2-fold; Supplementary Table S3 ). CERK is associated with ceramide metabolism through the effect of ceramide and sphingolipids on mitophagy . Another downregulated gene is GET1, which determines the kinetics of mitochondrial targeting proteins, therefore also controlling mitophagy . Surprisingly, among the downregulated genes ( Supplementary Table S2 ), many were assigned to neurons (NDNF, neuron-derived neurotrophic factor; NANOS1, nanos C2HC-type zinc finger 1) and synapses, including synaptic vesicle proteins (e.g., synaptophysin, SYP). We attribute this observation to the role of these genes in membrane trafficking. For example, syntaxin 6 (SNX6) functions as an endosomal organizer and intracellular protein transport. Another neuronal-like gene is SYT16, a calcium-independent membranous protein involved in the trafficking of secretory vesicles in non-neuronal tissues. Other genes were implicated in controlling neuronal survival and differentiation (e.g., GFRA1, a member of the glial cell-line-derived neurotrophic factor (GDNF) receptor family), and cellular adhesion (CNTNAP2, which mediates interactions between neurons and glia during development). Other genes belong to the TNF-receptor superfamily (e.g., TNFRS10D, TNFRSF21), indicative of their role in inflammation and in inhibition of cell apoptosis.
[ "Keren Zohar", "Michal Linial" ]
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2.6. Knockdown of PARK7/DJ-1 Exposed Downregulation of Genes with Membrane-Associated Functions
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[ 0.99951171875, 0.0002541542053222656, 0.00019633769989013672 ]
[ 0.99951171875, 0.00020778179168701172, 0.00034046173095703125, 0.00006651878356933594 ]
Figure 6 B indicates the set of genes that can be attributed to the KD of DJ-1 after filtering out the 206 upregulated overlapping genes (inset, Venn diagram). We focused on the 129 upregulated DJ-1 gene set . These genes were defined as significant (PDR < 0.05) among coding genes that showed a minimal log (FC) > 0.5). We identified STAT3 as a hub that connect stress, inflammation with transcription regulation (e.g., EGR1 and ATF3), and the DNA repair and cell cycle axis (CDKNA1 and DDB2). By eliminating the overwhelming signature of antiviral response, we could highlight the smaller gene set that was induced. As an example is the set of ANKRD36 family members, which was implicated in the pathology of immune and metabolic diseases , and considered as a potential diagnosis marker of Chronic myeloid leukemia .
[ "Keren Zohar", "Michal Linial" ]
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3.1. Cell Lines and siRNAs
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[ 0.99951171875, 0.0002663135528564453, 0.00021266937255859375 ]
[ 0.97998046875, 0.01873779296875, 0.0008764266967773438, 0.0003223419189453125 ]
Human neuroblastoma SH-SY5Y cells were purchased from ATCC (American Type Culture Collection, Rockville, MD, USA). Cells were cultured in Minimum Essential Media (MEM and F12 ratio 1:1, 4.5 g/L glucose) with 10% fetal calf serum (FCS) and 1:10 L-Alanyl-L-Glutamine. Cell cultures were incubated at 37 °C in a humidified atmosphere of 5% CO 2 .
[ "Keren Zohar", "Michal Linial" ]
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3.1. Cell Lines and siRNAs
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[ 0.99951171875, 0.00018668174743652344, 0.00021529197692871094 ]
[ 0.99853515625, 0.001262664794921875, 0.0003116130828857422, 0.0000680685043334961 ]
HEK293 cells were cultured to reach a 20–30% confluent level (3 × 10 4 per cm 2 ). For transfection, we used Lipofectamine 2000 in cells at 40–50% confluency according to the manufacturer’s instructions. Lipo2000 was shown to be ideal for HEK293 . The esiRNA (Mission system, Sigma-Aldrich, Burlington, MA, USA) consists of a pool of hundreds of siRNA (21 bp each), where each individual dsRNA has a low concentration in the pool, which diminishes most off-target effects, while producing an efficient knockdown. We apply the esiRNA-RULC of a mixture of 21 nt dsRNA used as a negative control (RLUC stands esiRNA directed against Renilla Luciferase; EHURLUC) in addition to the specific PARK7 . RULC experiments were used to measure the baseline effects of introducing esiRNA that controls for the delivery method and allows to distinguish the cellular response to the targeted siRNA itself .
[ "Keren Zohar", "Michal Linial" ]
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3.2. Reverse Transcription PCR (RT-PCR) and PCR
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[ 0.99951171875, 0.0002281665802001953, 0.00018477439880371094 ]
[ 0.9990234375, 0.0008101463317871094, 0.0002906322479248047, 0.00007534027099609375 ]
Cells were collected for RNA preparation. Total RNA was extracted from cell culture with TRIzol (Thermo-Fisher Scientific, Waltham, MA, USA), and RT was performed using a Ready-To-Go first-strand synthesis kit (Cytiva, Marlborough, MA, USA) according to the manufacturer’s instructions. RNA was reverse transcribed into cDNA (1 μg) and used in the PCR reaction. The PCR conditions consisted of denaturation at 95 °C for 2 min and 35 cycles (10 s at 95 °C, 15 s at 60 °C, 30 s at 72 °C), and 5 min for a final extension. The PCR products were separated on 1.5% agarose gel and stained with ethidium bromide, followed by densitometry measurement (using image processing ImageJ program Ver 1.54, from GitHub). The primers were designed against human RefSeq by the Primer3Web tool (Ver 4.1.0). The forward (F) and reverse (R) primers of β-actin were F: CATGCCCACCATCAGCCCTGG and R: ACAGAGCCTCGCCTTTGCCGA, respectively. For DJ-1 (376 nt), the primers were F: GCCTGGTGTGGGGCTTGTAA and R: GCCAACAGAGCAGTAGGACC. Amplicon size was confirmed to be 196 nt and 376 nt for β-actin and PARK7, respectively.
[ "Keren Zohar", "Michal Linial" ]
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3.3. RNA-Seq
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[ 0.99951171875, 0.00022470951080322266, 0.00019943714141845703 ]
[ 0.9931640625, 0.0060882568359375, 0.0005559921264648438, 0.00017559528350830078 ]
Total RNA was extracted using the RNeasy Plus Universal Mini Kit , according to the manufacturer’s protocol. Total RNA samples (1 μg RNA) were enriched for mRNAs by pull-down of poly (A). Libraries were prepared using a KAPA Stranded mRNA-Seq Kit, according to the manufacturer’s protocol, and sequenced using Illumina NextSeq 500 to generate 85 bp single-end reads (a total of 25–30 million reads per sample).
[ "Keren Zohar", "Michal Linial" ]
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3.4. Bioinformatic Analysis
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[ 0.99951171875, 0.00024819374084472656, 0.0001819133758544922 ]
[ 0.99853515625, 0.0010595321655273438, 0.0003428459167480469, 0.00010979175567626953 ]
Next-generation sequencing data underwent quality control using FastQC, version 0.11.9. They were then preprocessed using Trimmomatic (Ver. 0.32) and aligned to the reference genome GRCm38 with the STAR aligner (Ver. 2.7.0a) using default parameters. Genomic loci were annotated using GENCODE (release 46). Genes with low expression were filtered out of the dataset by setting a threshold of a minimum of two counts per million in three samples.
[ "Keren Zohar", "Michal Linial" ]
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3.4. Bioinformatic Analysis
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[ 0.99951171875, 0.00018155574798583984, 0.0002161264419555664 ]
[ 0.9990234375, 0.0004630088806152344, 0.00034928321838378906, 0.000055789947509765625 ]
Differential analyses were performed on all experimental groups, and genes with an FDR < 0.05 were considered. Only genes with an absolute log fold change of ≥0.5 were labeled up- or downregulated, all the rest were considered unchanged. We partition genes by type as coding and non-coding (including pseudogene, anti-sense, miRNAs, TEC, lncRNA, and other rare biotypes).
[ "Keren Zohar", "Michal Linial" ]
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3.4. Bioinformatic Analysis
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[ 0.99951171875, 0.0001666545867919922, 0.0002086162567138672 ]
[ 0.99951171875, 0.0004317760467529297, 0.00022554397583007812, 0.00005894899368286133 ]
RNA-seq experiment results are visualized by an MA plot that transforms the data into M (log ratio) and A (mean average) scales. The functional analysis and network view was based on STRING protein–protein interaction (PPI) platform. The minimal PPI confidence score ranges from 0.4 to 0.9. For protein interaction, we collected data from a human-centric view from IntAct, limiting DJ-1 interacting proteins to a minimal MI score > 0.6. Only evidence of physical interactions was included . We used the gene expression normalization nTPM to compare the expression of DJ-1 across cell types. Data are available in Human Proteome Atlas (HPA) .
[ "Keren Zohar", "Michal Linial" ]
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3.5. Statistical Analysis
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[ 0.99951171875, 0.0001392364501953125, 0.0005221366882324219 ]
[ 0.98095703125, 0.017791748046875, 0.0009822845458984375, 0.00015974044799804688 ]
Principal component analysis (PCA) was performed using the R-base function prcomp (R studio Ver. 4.1.0). EdgeR (Ver. 3.36.0) was used to perform RNA read counts by the trimmed mean of the M-values normalization of RNA (TMM) and differential expression analysis . Figures were generated using the ggplot2 R package (Ver.3.3.5) .
[ "Keren Zohar", "Michal Linial" ]
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4. Discussion
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[ 0.99951171875, 0.00023317337036132812, 0.0002148151397705078 ]
[ 0.99951171875, 0.00015020370483398438, 0.00043392181396484375, 0.00006031990051269531 ]
In this study, we investigated the role of KD in DJ-1 in the cellular context of HEK293 cells. We identified a robust and strong antiviral-like response that was also moderately induced by using the non-specific siRNA of RULC. The use of siRNA methodologies across different cell lines is a variable that is poorly controlled. Most commercially used siRNA protocols rely on a combination of a small set of specific sequences (about four), while the negative control setting uses scrambled sequences to ensure specificity. Several systematic studies indicated the importance of the cell-type-dependent nature of sensitivity to siRNA by its length and its impact on gene expression related to the innate immune response. For example, in HeLa cells, introducing dsRNAs longer than 23 bp induced toxicity and cell death, while shorter RNA fragments did not affect viability. In all cell types, short siRNA duplex (19 nt) did not affect cell viability .
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4. Discussion
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[ 0.99951171875, 0.00019109249114990234, 0.0001442432403564453 ]
[ 0.99853515625, 0.0004482269287109375, 0.0008091926574707031, 0.000072479248046875 ]
It is known that siRNA delivery reagents may induce stress in the subjected cells. Specifically, Lipo2000 is characterized by high transfection efficiency. As a byproduct of the use of Lipo2000 for siRNAs, autophagy is induced to cope with cellular oxidative and ER stress signals . We showed a strong induction of the antiviral-like response in HEK293 with no sign of cytotoxicity, and we still achieved effective silencing, retaining only 12% of the basal level of expression following 24–48 h post-transfection of siRNAs. The siRNA system and delivery are also sensitive to the presence of sequence motifs via the activation of toll-like receptors (e.g., TLR8) to induce an undesirable antiviral response . Due to the importance of siRNAs for research and as a promising therapeutic method, optimization is needed to achieve the desired responses while optimizing silencing efficiency.
[ "Keren Zohar", "Michal Linial" ]
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4. Discussion
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[ 0.99951171875, 0.0002574920654296875, 0.00021731853485107422 ]
[ 0.99951171875, 0.0003590583801269531, 0.000274658203125, 0.00007826089859008789 ]
Similar to our finding, siRNAs (21 nt) that were introduced into cells for silencing specific genes, triggered the JAK/STAT signaling pathway . The signaling cascade was initiated by EIF2AK2 (PKR, an IFN-induced dsRNA-dependent kinase), which plays a central role in the innate immune response to viral infection . In our experiments, EIF2AK2 was already overexpressed (four-fold) by the siRNA of the RULC, which is likely to mediate the observed antiviral-like response. The involvement of trafficking and organelle biogenesis in the antiviral response is established, along with the executing of INF signaling .
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277157_p29
PMC11277157
sec[3]/p[3]
4. Discussion
4.222656
biomedical
Study
[ 0.99951171875, 0.00021278858184814453, 0.00011521577835083008 ]
[ 0.998046875, 0.0003376007080078125, 0.0013179779052734375, 0.00011962652206420898 ]
It was shown that DJ-1 deficiency or knockdown in primary cortical neurons and mouse embryonic fibroblasts caused alternations in mitochondrial shape . Knockout mice and DJ-1 pathogenic mutations led to elevated ROS production, lower mitochondrial membrane potential, and changes in gene expression, which can thus explain the importance of DJ-1 function in neurodegenerative diseases . These in vivo findings corroborate the observed effects of suppressing DJ-1 on trafficking, mitophagy, and redox homeostasis .
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
PMC11277157_p30
PMC11277157
sec[3]/p[4]
4. Discussion
4.492188
biomedical
Study
[ 0.99951171875, 0.0004811286926269531, 0.00021767616271972656 ]
[ 0.9990234375, 0.0003731250762939453, 0.0005679130554199219, 0.00017940998077392578 ]
We observed that the antiviral effect was strongly amplified by the KD of DJ-1 (with ~200 overexpressed genes and ~100 non-coding RNAs). Most of the ncRNA DEGs are expressed in a moderate-to-low level . While no function is known for most of these non-coding RNAs, the upregulation of lncRNA of NEAT1 (2.2-fold, q -value 3.7 × 10 −5 ) is intriguing, as NEAT1 was proposed to be upregulated in response to oxidative stress and as a potential biomarker in Parkinson’s diseases . We questioned whether the effect of the KD of DJ-1 is through the signaling convergence of the IFN signaling pathways or, alternatively, through the failure of the cells to cope with the lack of DJ-1 protection and changes in the redox level. We suggest that it is the former explanation. We have not challenged the cells with an oxidative stimulus , and, as anticipated, no sign of cytotoxicity following siRNAs was observed, which argues for uninterrupted mitochondrial homeostasis. In contrast, the functional networks presented in this study indicate the key role of JAK/STAT as the main hub. DJ-1 negatively regulates inflammatory responses in astrocytes and microglia by facilitating interactions between STAT1 and its phosphatase, SHP-1 . Experiments performed on DJ-1 knockout mice led to increased inflammatory mediators compared to wild-type mice. This was explained by the direct interaction of DJ-1 with SHP-1 and shifting the levels of phosphorylated STAT1. Therefore, DJ-1 may prevent excessive STAT1 activation and reduce the risk of brain inflammation . It remains to be tested whether in our cell system, the activation of the strong antiviral response in KD DJ-1 cells is mediated through such a signaling pathway.
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277157_p31
PMC11277157
sec[3]/p[5]
4. Discussion
4.144531
biomedical
Study
[ 0.99951171875, 0.00024056434631347656, 0.0001837015151977539 ]
[ 0.99951171875, 0.00017940998077392578, 0.0003192424774169922, 0.00006639957427978516 ]
In this study, we tested the KD of DJ-1 in a non-cancerous established cell line by inspecting the transcriptional response. We emphasized that different cells present varying sensitivity to molecular perturbations and external stimuli . We argue that the depletion of DJ-1 in HEK293 cells caused a slight alteration in membranous trafficking but mostly a change in transcription, rendering the cells labile and prone to multiple stressors, including dsRNA-INF-dependent viral-like stress. Our study highlights the intricate interplay between siRNA technology and the robust activation of the innate immune system.
[ "Keren Zohar", "Michal Linial" ]
https://doi.org/10.3390/ijms25147550
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
39057620_p0
39057620
sec[0]/p[0]
1. Introduction
4.132813
biomedical
Clinical case
[ 0.767578125, 0.231201171875, 0.0012788772583007812 ]
[ 0.10821533203125, 0.058837890625, 0.01293182373046875, 0.81982421875 ]
Recent studies have demonstrated the benefits of performing complex percutaneous coronary interventions (PCIs) with the support of intracoronary imaging techniques, and international clinical guidelines recommend their use . Optical Coherence Tomography (OCT) uses near-infrared light to obtain in vivo images with high spatial resolution, enabling detailed examination of the interior of coronary vessels. This makes it an invaluable tool for the diagnosis, planning and optimization of PCI in complex scenarios. While complications are rare, the occurrence of ventricular arrhythmias during its use is one of the most feared. Unfortunately, the mechanism by which these arrhythmias arise remains unclear. Understanding the mechanism of their occurrence is essential for prevention. Here, we discuss a case of OCT-guided PCI during which ventricular fibrillation occurred.
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
39057620_p1
39057620
sec[1]/p[0]
2. Detailed Case Description
3.929688
clinical
Clinical case
[ 0.072509765625, 0.923828125, 0.0037899017333984375 ]
[ 0.00605010986328125, 0.00785064697265625, 0.005275726318359375, 0.98095703125 ]
A 60-year-old male, current smoker and dyslipidemic, was admitted to the emergency room due to chest pain. He referred to several episodes of chest pain on slight exertion in the last 48 h. His vitals revealed a temperature of 36.5 °C, a heart rate of 55 beats per minute, blood pressure of 130/78 mmHg and oxygen saturation of 99% on room air. A physical examination did not reveal either cardiac murmurs or signs of congestive heart failure. The initial ECG showed sinus rhythm with first-degree atrioventricular block, without repolarization alterations . The transthoracic echocardiogram showed a normal left-ventricular ejection fraction without segmental alterations of contractility and no valvular disease. Initial laboratory reports revealed a slight elevation of cardiac damage markers (ultrahigh-sensitivity troponin I of 158 ng/L [reference range level, 0.0–54.0 ng/L]), with the rest of the analytical parameters in the normal range. With the diagnosis of non-ST-elevation acute coronary syndrome, 300 mg of acetylsalicylic acid and 180 mg of Ticagrelor were administered and the patient was admitted to Cardiology with telemetry. The following morning, the patient underwent a diagnostic coronary angiography via the right radial artery, which revealed a critical stenosis in the ostium of the left anterior descending artery (LAD) and an intermediate stenosis in its middle segment, with diffuse disease between both segments (TIMI III flow). No other significant stenosis was observed in the rest of the coronary vessels .
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
39057620_p2
39057620
sec[1]/p[1]
2. Detailed Case Description
3.248047
clinical
Clinical case
[ 0.2430419921875, 0.75439453125, 0.0025310516357421875 ]
[ 0.01470947265625, 0.05169677734375, 0.0021514892578125, 0.931640625 ]
The patient was informed of the findings and the various treatment options. It was decided to perform PCI in the same procedure guided by intracoronary OCT imaging to assess the anatomy, take measurements and evaluate the possible involvement of the distal left main coronary artery (LMA) as well as the ostium of the LAD and circumflex artery (CX).
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999999
39057620_p3
39057620
sec[1]/p[2]
2. Detailed Case Description
3.982422
clinical
Clinical case
[ 0.385498046875, 0.61181640625, 0.0025234222412109375 ]
[ 0.11798095703125, 0.368408203125, 0.0059661865234375, 0.5078125 ]
Intravenous unfractionated heparin (100 U per kg) was administered. The LMA was catheterized with a 6F 4 EBU guiding catheter, then a Sion guidewire was advanced to the distal segment of the LAD, and a Sion blue guidewire was advanced to the distal segment of the CX. Subsequently, the ostial stenosis of the LAD was dilated using a semi-compliant balloon with a diameter of 2.5 mm. The OCT catheter was then advanced to the mid-distal segment of the LAD, and a first pullback was performed by injecting 16 mL of contrast (4 mL/s for 4 s) without achieving clear intracoronary images. Subsequently, a second pullback was performed, and 29 mL of contrast (5 mL/s for 6 s) were injected through the guide catheter, acquiring optimal images .
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999998
39057620_p4
39057620
sec[1]/p[3]
2. Detailed Case Description
3.964844
clinical
Clinical case
[ 0.09576416015625, 0.9013671875, 0.00284576416015625 ]
[ 0.01358795166015625, 0.022918701171875, 0.003871917724609375, 0.95947265625 ]
Immediately after completing the injection, the patient experienced ventricular fibrillation (VF) , requiring external defibrillation with 360 J, which successfully restored sinus rhythm. The patient recovered promptly. Based on the OCT images obtained from the prior OCT, we proceed with the deployment of a 3.5/18 mm sirolimus-eluting stent from the proximal LAD to the ostium of the LMA, followed by postdilatation of the proximal segment with a 4.5 mm non-compliant balloon at 16 atm. Additionally, the stent struts were opened towards the ostium of the CX using a 2.5 mm non-compliant balloon, concluding with a LAD-CX kissing balloon inflation. Finally, a 2.75/20 mm sirolimus-eluting stent was implanted at 20 atm in the intermediate stenosis of the middle segment of the LAD, which had a minimum luminal area of 2.1 mm 2 .
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
39057620_p5
39057620
sec[1]/p[4]
2. Detailed Case Description
3.689453
clinical
Clinical case
[ 0.303955078125, 0.6943359375, 0.001953125 ]
[ 0.031646728515625, 0.03155517578125, 0.002593994140625, 0.93408203125 ]
A new OCT, injecting 15 mL of contrast (4 mL/s for 4 s), revealed adequate apposition and expansion of the LMA stent, with no evidence of distal or proximal dissection, and it also confirmed the correct opening of the stent struts at the ostium of the CX . The patient did not experience any further ventricular arrhythmias during this last OCT.
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
39057620_p6
39057620
sec[1]/p[5]
2. Detailed Case Description
1.274414
clinical
Clinical case
[ 0.0296478271484375, 0.9599609375, 0.01047515869140625 ]
[ 0.002544403076171875, 0.018035888671875, 0.002475738525390625, 0.97705078125 ]
During admission, the patient progressed without complications and was discharged 24 h after catheterization without any further events.
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999999
39057620_p7
39057620
sec[2]/p[0]
3. Discussion
4.0625
biomedical
Study
[ 0.9990234375, 0.0006809234619140625, 0.0001577138900756836 ]
[ 0.98095703125, 0.0015573501586914062, 0.0171966552734375, 0.0002532005310058594 ]
OCT has been evaluated for the assessment of intermediate stenosis . Several studies have recently been published demonstrating the benefit of performing complex PCI with the support of various intracoronary imaging techniques. The multicenter OCTOBER trial showed that using OCT during PCI of bifurcation lesions reduces adverse cardiovascular events at 2-year follow-up compared to PCI performed with angiography alone . While intravascular ultrasound (IVUS) has been the traditional method for evaluating LMA lesions, the OCTOBER trial results support the emerging role of OCT as a valuable tool for the assessment of this lesions, as nearly 20% of the patients had a bifurcation lesion involving the LMA .
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
39057620_p8
39057620
sec[2]/p[1]
3. Discussion
4.089844
biomedical
Study
[ 0.9853515625, 0.01447296142578125, 0.00030159950256347656 ]
[ 0.68798828125, 0.26708984375, 0.00861358642578125, 0.036468505859375 ]
The OCT catheter captures high-resolution images of the inside of the artery by emitting a beam of light . Contrary to IVUS, OCT requires the injection of intracoronary contrast to displace the blood in order to obtain clear images of the vessel. The amount of contrast needed depends on the diameter and length of the segment to be studied, as well as the speed of blood flow. Therefore, there is no fixed guideline for the amount of contrast to be administered. In general, it is recommended to inject 15–20 mL at a rate of 4 mL/second to acquire a pullback in the left coronary tree . However, frequently, this amount of contrast is inadequate to displace the blood from the vessel, resulting in artifacts and making image interpretation challenging. As a result, it is often necessary to repeat acquisitions and substantially increase the final total amount of contrast administered, as occurred in our case, where we had to administer 29 mL to adequately characterize the disease from the middle segment of the LAD to the LMA (55 mm in length).
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999999
39057620_p9
39057620
sec[2]/p[2]
3. Discussion
4.035156
biomedical
Study
[ 0.98583984375, 0.013427734375, 0.0004973411560058594 ]
[ 0.97607421875, 0.02069091796875, 0.002147674560546875, 0.0009293556213378906 ]
One of the main concerns about OCT is the higher use of contrast, with the consequent risk of deterioration of renal function. However, in the OPINION trial, which compared OCT-guided PCI with IVUS-guided PCI, although the total amount of contrast was higher in the OCT group (400 patients), none of them suffered from contrast-induced nephropathy . Moreover, in the OCTOBER trial, only one patient in the OCT group suffered acute deterioration of renal function after PCI .
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
39057620_p10
39057620
sec[2]/p[3]
3. Discussion
4.054688
biomedical
Study
[ 0.99853515625, 0.0010986328125, 0.00012671947479248047 ]
[ 0.9912109375, 0.006443023681640625, 0.0019474029541015625, 0.0005292892456054688 ]
From an electrical standpoint, infrequent effects have been described, such as T-wave inversion, ST-segment depression or J-point elevation . Complex ventricular arrhythmias are exceptionally rare, for example, in the OCTOBER trial, only three cases of ventricular tachycardia (VT) or VF were reported among the 600 patients who underwent PCI with OCT, without describing the mechanisms that triggered them .
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
39057620_p11
39057620
sec[2]/p[4]
3. Discussion
4.234375
biomedical
Study
[ 0.95361328125, 0.045654296875, 0.0004851818084716797 ]
[ 0.64990234375, 0.056793212890625, 0.00736236572265625, 0.2861328125 ]
In our case, the electrocardiographic recording during contrast injection revealed a progressive ST-segment elevation and a progressive widening of the QRS complex as the contrast was injected into the LMA. Subsequently, a short-coupled extrasystole triggered polymorphic VT that rapidly degenerated into VF , as described by Terada et al. in all their patients . ST elevation and QRS prolongation are described as markers of myocardial ischemia during PCI, with QRS widening being an even more sensitive marker than ST segment changes. This alteration is more pronounced when proximal and middle segments of the main arteries are occluded . The underlying proarrhythmic mechanism of this R-on-T phenomenon has been described in the context of bradycardia, hypoxia, certain drug use and the presence of hydroelectrolyte disturbances .
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
39057620_p12
39057620
sec[2]/p[5]
3. Discussion
3.947266
biomedical
Study
[ 0.99755859375, 0.00238800048828125, 0.00013315677642822266 ]
[ 0.98779296875, 0.0087127685546875, 0.0010843276977539062, 0.0025157928466796875 ]
Myocardial ischemia could potentially be attributed to other causes reported in the literature, such as coronary artery spasm, deep intubation of the guiding catheter, air embolization or even dissection/plaque rupture with coronary occlusion during OCT pullback. However, all these possibilities were excluded by the simultaneous angiography done during OCT pullback ( Video S2 ) and in subsequent OCT analysis ( Video S3 ).
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
39057620_p13
39057620
sec[2]/p[6]
3. Discussion
3.517578
biomedical
Clinical case
[ 0.697265625, 0.30078125, 0.0019245147705078125 ]
[ 0.053802490234375, 0.11962890625, 0.002567291259765625, 0.82421875 ]
Therefore, we believe that the primary cause of the arrhythmia in our patient was myocardial ischemia, which may be attributed to the significant volume of contrast utilized during the second pullback. The posterior R-on-T phenomenon was a key factor in precipitating the subsequent VF.
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
39057620_p14
39057620
sec[2]/p[7]
3. Discussion
3.412109
biomedical
Study
[ 0.74560546875, 0.253173828125, 0.0014905929565429688 ]
[ 0.39697265625, 0.308349609375, 0.002201080322265625, 0.29248046875 ]
After defibrillation, the procedure was successfully completed, and a post-PCI control OCT was performed. As we only intended to assess the treated segment in LAD and LMCA (with a total length of 30 mm), we only used half the initial contrast volume, without reproducing any ventricular arrhythmia.
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
39057620_p15
39057620
sec[3]/p[0]
4. Conclusions
4.082031
clinical
Clinical case
[ 0.29833984375, 0.6982421875, 0.00328826904296875 ]
[ 0.018829345703125, 0.04144287109375, 0.00460052490234375, 0.93505859375 ]
To the best of our knowledge, this is the first reported case of VF during OCT preceded by widening of the QRS complex and ST elevation, suggesting myocardial ischemia as the mechanism favoring VF. With high probability, the amount of contrast administered in the second pullback was the determinant of VF. For this reason, it is recommended to adjust the volume of contrast injected during OCT acquisition, seeking in each case the balance between image quality and contrast volume. Possible electrical changes in telemetry in the first seconds after OCT should be monitored closely, and a defibrillator should always be available so that rapid action can be taken if ventricular arrhythmia occurs.
[ "Paula Vela Martín", "Carlos Arellano Serrano", "Álvaro Lorente Ros", "Juan Francisco Oteo", "Arturo Garcia-Touchard" ]
https://doi.org/10.3390/jcdd11070200
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996
PMC11277181_p0
PMC11277181
sec[0]/p[0]
Introduction
4.289063
biomedical
Review
[ 0.998046875, 0.0015726089477539062, 0.00044798851013183594 ]
[ 0.1719970703125, 0.138671875, 0.6787109375, 0.0103912353515625 ]
Waardenburg syndrome (WS) represents an autosomal dominant genetic disorder typified by the absence of melanocytes in the eyes, hair, and skin structures , leading to its classification as an auditory-pigmentary syndrome. Clinically, it is delineated by hallmark pigmentary disturbances of the hair, notably the presence of a white forelock and premature graying; ocular pigmentary discrepancies, including heterochromia irides and brilliant blue irises; in conjunction with congenital sensorineural hearing loss . The global incidence of WS is estimated at two per 100,000 individuals .
[ "Ameer Awashra", "Ahmad Nouri", "Thabet Zidan", "Abdelrahman Sawalma", "Fathi S Milhem", "Dalia Marbo’" ]
https://doi.org/10.7759/cureus.63206
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999997
PMC11277181_p1
PMC11277181
sec[0]/p[1]
Introduction
3.296875
clinical
Clinical case
[ 0.2261962890625, 0.767578125, 0.006008148193359375 ]
[ 0.0028438568115234375, 0.007617950439453125, 0.00257110595703125, 0.98681640625 ]
We report a preterm male newborn with a predisposition to WS, who presented with multiple complications at birth. Despite early respiratory and feeding challenges, he stabilized with no neurological issues. Genetic testing confirmed WS and multisystem anomalies were managed effectively. The infant’s recovery was supported by a multidisciplinary team, ensuring a successful transition to regular oral feeding and ongoing specialized care.
[ "Ameer Awashra", "Ahmad Nouri", "Thabet Zidan", "Abdelrahman Sawalma", "Fathi S Milhem", "Dalia Marbo’" ]
https://doi.org/10.7759/cureus.63206
N/A
https://creativecommons.org/licenses/by/4.0/
en
0.999996