Correlation of synovial caspase-3 concentration and the photodynamic effectiveness in osteoarthritis treatment
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01.06.2020 |
Zharova T.
Kogan E.
Makarov V.
Smorchkov M.
Lychagin A.
Ivannikov S.
Zharkov N.
Loschenov V.
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Photodiagnosis and Photodynamic Therapy |
10.1016/j.pdpdt.2020.101669 |
0 |
Ссылка
© 2020 Elsevier B.V. Background: The present study focuses on investigation of Intra-articular PDT mechanisms for OA treatment. Also, a search for determination of the most effective dose of chlorin e6 (Ce6) for anti-inflammatory PDT of OA was carried out. Methods: The study was carried out on laboratory animals (11 Chinchilla rabbits, 1 year, 2.5 kg) with a gonarthritis model of post-traumatic OA. According to the instructions for using Photoditazin (Ce6 based PS) for PDT of human oncological and non-oncological diseases, the recommended dose is 0.7–1.2 mg/kg. For studies on rabbits, taking into account the conversion coefficient (3.2), the PS doses of 2.4, 3.2 and 6.4 mg/kg were selected. Fluorescence spectra were measured intra-articular before and after PDT using spectrometer with fiber-optic probe. The intrajoint PDT was carried out using a laser (662 ± 10 nm) and a fiber-optic catheter with a cylindrical diffuser inside a sapphire needle for a uniform distribution of the laser radiation. The immunohistochemical study was carried out by staining the samples with caspase-3. Results: Histological and immunohistochemical analysis showed that the best PS dose for intravenous administration for PDT of rabbit gonarthritis is 3.2 mg/kg. The PS concentration directly in the synovial tissue was 0.5 mg/kg, and this was enough to achieve the most positive results to reduce the caspase-3 level. Conclusion: The caspase-3 level correlates well with other signs of inflammation in the synovial membrane (edema, etc.). Therefore, to assess the PDT effectiveness in the treatment of gonarthritis accompanied by synovitis, it is sufficient to analyze only for caspase-3. The efficacy of PDT with Ce6 showed that 3.2 mg/kg PS dose (1 mg/kg for a human) is the most effective.
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Correlation of synovial caspase-3 concentration and the photodynamic effectiveness in osteoarthritis treatment
|
01.06.2020 |
Zharova T.
Kogan E.
Makarov V.
Smorchkov M.
Lychagin A.
Ivannikov S.
Zharkov N.
Loschenov V.
|
Photodiagnosis and Photodynamic Therapy |
10.1016/j.pdpdt.2020.101669 |
0 |
Ссылка
© 2020 Elsevier B.V. Background: The present study focuses on investigation of Intra-articular PDT mechanisms for OA treatment. Also, a search for determination of the most effective dose of chlorin e6 (Ce6) for anti-inflammatory PDT of OA was carried out. Methods: The study was carried out on laboratory animals (11 Chinchilla rabbits, 1 year, 2.5 kg) with a gonarthritis model of post-traumatic OA. According to the instructions for using Photoditazin (Ce6 based PS) for PDT of human oncological and non-oncological diseases, the recommended dose is 0.7–1.2 mg/kg. For studies on rabbits, taking into account the conversion coefficient (3.2), the PS doses of 2.4, 3.2 and 6.4 mg/kg were selected. Fluorescence spectra were measured intra-articular before and after PDT using spectrometer with fiber-optic probe. The intrajoint PDT was carried out using a laser (662 ± 10 nm) and a fiber-optic catheter with a cylindrical diffuser inside a sapphire needle for a uniform distribution of the laser radiation. The immunohistochemical study was carried out by staining the samples with caspase-3. Results: Histological and immunohistochemical analysis showed that the best PS dose for intravenous administration for PDT of rabbit gonarthritis is 3.2 mg/kg. The PS concentration directly in the synovial tissue was 0.5 mg/kg, and this was enough to achieve the most positive results to reduce the caspase-3 level. Conclusion: The caspase-3 level correlates well with other signs of inflammation in the synovial membrane (edema, etc.). Therefore, to assess the PDT effectiveness in the treatment of gonarthritis accompanied by synovitis, it is sufficient to analyze only for caspase-3. The efficacy of PDT with Ce6 showed that 3.2 mg/kg PS dose (1 mg/kg for a human) is the most effective.
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Apoptosis in seminiferous tubules of human in normal and in idiopathic infertility
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01.01.2018 |
Demyashkin G.
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Tsitologiya |
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1 |
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© 2018 Sankt Peterburg. All rights reserved. The purpose of this study was to assess the level of apoptosis in cells of human seminiferous tubules in normal and pathological spermatogenesis by revealing the proportion of immunostaining cells for caspase-9 and -3 and by comparing the expression of pro-apoptotic (BAK and BAX) and anti-apoptotic genes (BCL2 and BCLW). A retrospective study involved men (n = 42) who complained of childlessness in marriage for 2 years with a diagnosis of idiopathic infertility, established after physical, genetic, biochemical (hormones) and cytological (spermogram) analyzis. The biopsies of the testes as well as the autopsy material of men 22—35 (n = 10), 64—75 (n = 10) and 75—90 (n = 10) years were studied using the immunohistochemical method (caspase-9 and -3) and PCR-RT. The marking level for caspase-9 in spermatogonies with normal spermatogenesis is approximately at the same level (the proportion of stained spermatogonies is 39.5 0.33 % in young men, 35.6 0.44 % in the elderly, and 32.2 0.28 % in man of senile age), and increases when the maturation is blocked and in the case of focal variant of Sertoli-cell-only syndrome (64.3 0.39 and 72.0 0.41 %, respectively). When using antibodies to caspase-3, the percentage of immunopositive spermatogonia in normal spermatogenesis was 60.1 0.44 % in young men, 78.2 1.2 % in the elderly, and 87.3 0.9 % in men of senille age, and with idiopathic infertility, a sharp increase in the proportion of labeled spermatogonia was observed (an average of 91.4 1.1 %). In the case of hypospermatogenesis and the blocking of maturation, a significant increase in the relative level of expression of the proapoptotic genes of the internal pathway of apoptosis of BAX and BAK was observed against the background of a decrease in the expression of the anti-apoptotic genes BCL2 and BCLW, compared to men of the same age but with normal spermatogenesis. Based on the results, we can conclude that in the elderly’s testes the receptor-mediated pathway of apoptosis predominates over the mitochondrial (internal) pathway. In the idiopathic form of male infertility the internal pathway of apoptosis is dominant. An increase in the activity of apoptosis markers can be associated with impaired maturation of the germ cells in the meiosis block and with the depletion of the germ cell pool in Sertoli-cell-only syndrome.
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