Effect of Chaerophyllum macropodum extracts on Trichomonas vaginalis in vitro

Introduction: Trichomonas vaginalis (T. vaginalis) is a protozoan parasite causing trichomoniasis or trichomonal vaginitis. The infection is considered as non-viral sexually transmitted disease (STD). Metronidazole and Tinidazole are now the drugs of choice for the treatment of this infection. However, resistant to these drugs has also been reported. So it is necessary to search for effective alternative drugs with fewer side effects. Chaerophyllum macropodum (C. macropodum) plant have been used against some parasites. Therefore, in this study the effects of different extracts of this plant on T. vaginalis in culture media have been investigated. Methods: In this experimental study hydro-ethanol extracts of C. macropodum leaves were prepared. Anti-T. vaginalis activities of the extracts were tested in concentrations of 2, 4, 8, 16, 32, 40, 50, 60, 80, 100 and 150 mg/ml following 24, 48 and 72 hours of incubation of cultured media. Results: All extract concentrations showed some degrees of growth inhibition activity on T. vaginalis. However crude extract was more efficient. Conclusion: C. macropodum showed an anti-T. vaginalis activity. More investigations are recommended to use this plant as an antiparasitic drug

Effect of Chaerophyllum macropodum extracts on Trichomonas vaginalis in vitro

Introduction: Trichomonas vaginalis (T. vaginalis) is a protozoan parasite causing trichomoniasis or trichomonal vaginitis. The infection is considered as non-viral sexually transmitted disease (STD). Metronidazole and Tinidazole are now the drugs of choice for the treatment of this infection. However, resistant to these drugs has also been reported. So it is necessary to search for effective alternative drugs with fewer side effects. Chaerophyllum macropodum (C. macropodum) plant have been used against some parasites. Therefore, in this study the effects of different extracts of this plant on T. vaginalis in culture media have been investigated. Methods: In this experimental study hydro-ethanol extracts of C. macropodum leaves were prepared. Anti-T. vaginalis activities of the extracts were tested in concentrations of 2, 4, 8, 16, 32, 40, 50, 60, 80, 100 and 150 mg/ml following 24, 48 and 72 hours of incubation of cultured media. Results: All extract concentrations showed some degrees of growth inhibition activity on T. vaginalis. However crude extract was more efficient. Conclusion: C. macropodum showed an anti-T. vaginalis activity. More investigations are recommended to use this plant as an antiparasitic drug

Differentiation of human primary testicular cells in the presence of SCF using the organoid culture system

Purpose: Development of organoids using human primary testicular cells has remained a challenge due to the complexity of the mammalian testicular cytoarchitecture and culture methods. In this study, we generated testicular organoids derived from human primary testicular cells. Then, we evaluated the effect of stem cell factor (SCF) on cell differentiation and apoptosis in the testicular organoid model. Methods: The testicular cells were harvested from the three brain-dead donors. Human spermatogonial stem cells (SSCs) were characterized using immunocytochemistry (ICC), RT-PCR and flow cytometry. Testicular organoids were generated from primary testicular cells by hanging drop culture method and were cultured in three groups: control group, experimental group 1 (treated FSH and retinoic acid (RA)), and experimental group 2 (treated FSH, RA and SCF), for five weeks. We assessed the expression of SCP3 (Synaptonemal Complex Protein 3) as a meiotic gene, PRM2 (Protamine 2) as a post-meiotic marker and apoptotic genes of Bax (BCL2-Associated X Protein) and Bcl-2 (B-cell lymphoma 2), respectively by using RT-qPCR. In addition, we identified the expression of PRM2 by immunohistochemistry (IHC). Results: Relative expression of SCP3, PRM2 and Bcl-2 were highest in group 2 after five weeks of culture. In contrast, BAX expression level was lower in experimental group 2 in comparison with other groups. IHC analyses indicated the highest expression of PRM2 as a postmeiotic marker in group 2 in comparison to 2D culture and control groups but not find significant differences between experimental group 1 and experimental group 2 groups. Morphological evaluations revealed that organoids are compact spherical structures and in the peripheral region composed of uncharacterized elongated fibroblast-like cells. Conclusion: Our findings revealed that the testicular organoid culture system promote the spermatogonial stem cell (SSC) differentiation, especially in presence of SCF. Developed organoids are capable of recapitulating many important properties of a stem cell niche

The germ cell specific markers ZPBP2 and PGK2 in testicular biopsies can predict the presence as well as the quality of sperm in non-obstructive azoospermia patients

To assess the role of three testis-specific genes including ZPBP2, PGK2, and ACRV1 in the prediction of sperm retrieval result and quality of retrieved sperm by microdissection testicular sperm extraction (micro-TESE) in non-obstructive azoospermia (NOA) patients. This was a case-control study including 57 testicular samples of NOA patients including 32 patients with successful sperm retrieval (NOA+) and 25 patients with failed sperm retrieval (NOA-), and 9 samples of men with normal spermatogenesis in the testes as the positive control (OA). We investigated the expression of candidate genes by RT-qPCR and germ cell population patterns by DNA flow cytometry in testicular biopsy samples. The association between PGK2 expressions with the quality of retrieved spermatozoa was also evaluated. The RT-qPCR data revealed a significantly higher expression of ZPBP2 and PGK2 in the NOA+ in comparison to NOA- group (P = 0.002, and P = 0.002, respectively). Flow cytometry results revealed that the haploid cell percentage was significantly higher in NOA+ vs. NOA- group (P = 0.0001). In samples with a higher percentage of haploid cells, expression levels of ZPBP2 and PGK2 were higher (P = 0.001). The PGK2 expression was significantly associated with retrieved sperm quality (P = 0.01). Our results contribute to the search for the biomarkers for predicting the presence of testicular sperm and would be useful to avoid unnecessary multiple micro-TESE. Overall, the expression pattern of the ZPBP2 and PGK2 may be useful in predicting sperm recovery success and quality of retrieved sperm in NOA patients.</p

In vitro complete differentiation of human spermatogonial stem cells to morphologic spermatozoa using a hybrid hydrogel of agarose and laminin

Spermatogenesis refers to the differentiation of the spermatogonial stem cells (SSCs) located in the base seminiferous tubules into haploid spermatozoa. Prerequisites for in vitro spermatogenesis include an extracellular matrix (ECM), paracrine factors, and testicular somatic cells which play a supporting role for SSCs. Thus, the present study evaluated the potential of co-culturing Sertoli cells and SSCs embedded in a hybrid hydrogel of agarose and laminin, the main components of the ECM. Following the three–week conventional culture of human testicular cells, the cells were cultured in agarose hydrogel or agarose/laminin one (hybrid) for 74 days. Then, immunocytochemistry, real-time PCR, electron microscopy, and morphological staining methods were applied to analyze the presence of SSCs, as well as the other cells of the different stages of spermatogenesis. Based on the results, the colonies with positive spermatogenesis markers were observed in both culture systems. The existence of the cells of all three phases of spermatogenesis (spermatogonia, meiosis, and spermiogenesis) was confirmed in the two groups, while morphological spermatozoa were detected only in the hybrid hydrogel group. Finally, a biologically improved 3D matrix can support all the physiological activities of SSCs such as survival, proliferation, and differentiation

Differentiation of human primary testicular cells in the presence of SCF using the organoid culture system

Purpose: Development of organoids using human primary testicular cells has remained a challenge due to the complexity of the mammalian testicular cyto-architecture and culture methods. In this study, we generated testicular organoids derived from human primary testicular cells. Then, we evaluated the effect of stem cell factor (SCF) on cell differentiation and apoptosis in the testicular organoid model.Methods: The testicular cells were harvested from the three brain-dead donors. Human spermatogonial stem cells (SSCs) were characterized using immunocytochemistry (ICC), RT-PCR and flow cytometry. Testicular organoids were generated from primary testicular cells by hanging drop culture method and were cultured in three groups: control group, experimental group 1 (treated FSH and retinoic acid (RA)), and experimental group 2 (treated FSH, RA and SCF), for five weeks. We assessed the expression of SCP3 (Synaptonemal Complex Protein 3) as a meiotic gene, PRM2 (Protamine 2) as a post-meiotic marker and apoptotic genes of Bax (BCL2-Associated X Protein) and Bcl-2 (B-cell lymphoma 2), respectively by using RT-qPCR. In addition, we identified the expression of PRM2 by immunohistochemistry (IHC).Results: Relative expression of SCP3, PRM2 and Bcl-2 were highest in group 2 after five weeks of culture. In contrast, BAX expression level was lower in experimental group 2 in comparison with other groups. IHC analyses indicated the highest expression of PRM2 as a postmeiotic marker in group 2 in comparison to 2D culture and control groups but not find significant differences between experimental group 1 and experimental group 2 groups. Morphological evaluations revealed that organoids are compact spherical structures and in the peripheral region composed of uncharacterized elongated fibroblast-like cells.Conclusion: Our findings revealed that the testicular organoid culture system promote the spermatogonial stem cell (SSC) differentiation, especially in presence SCF. Developed organoids are capable of recapitulating many important prop?erties of a stem cell niche.</p

The impact of COVID-19 pandemic on depression and anxiety symptoms: Findings from the United Arab Emirates Healthy Future (UAEHFS) cohort study.

BACKGROUND: Significant concerns about mental health were raised during the COVID-19 pandemic. We investigated the prevalence of depression and anxiety symptoms among the participants of the United Arab Emirates Healthy Future Study (UAEHFS); a national cohort study. We further explored the change in the prevalence of depression symptoms among those with comparable pre-pandemic data. METHODS: A sample of UAEHFS participants were invited to complete a COVID-19 online questionnaire during the first wave of the pandemic. Depression and anxiety symptoms were assessed using the Patient Health Questionnaire Depression Scale (PHQ-8) and the Generalized Anxiety Disorder-7 Scale (GAD-7) respectively. Unpaired analyses were done to examine the effect of COVID-19 on depression and anxiety symptoms during the pandemic. Paired analysis was conducted to examine the change in depression symptoms. RESULTS: During the pandemic, we reported a prevalence of 32.8% (95% CI: 27.0, 39.1) for depression and 26.4% (95% CI: 21.0, 32.6) for anxiety symptoms. Younger people reported higher levels of depression (40.4%) and anxiety (34.5%) symptoms. Females reported higher levels of depression (36.5%) and anxiety (32.7%) symptoms. In paired analysis, the prevalence of depression symptoms during the pandemic was 34% (95% CI: 26.5, 42.4) compared to 29.9% (95% CI: 22.7, 38.1) before the pandemic. No statistically significant difference was observed, p-value = 0.440. Adjusted multivariate logistic regression models for PHQ-8 and GAD-7 during the pandemic showed that participants, who were experiencing flu-like symptoms, had higher odds of reporting depression symptoms compared to those without symptoms. Additionally, age was significantly negatively associated with anxiety symptoms. CONCLUSIONS: Overall, we found that depression and anxiety symptoms were more prevalent among young people and females. However, we did not find a significant change in the prevalence of depression symptoms among those with comparable pre-pandemic data. Identifying vulnerable groups and understanding trajectories through longitudinal studies would help with planning for effective mental health interventions for the current and future pandemics

The lipid elongation enzyme ELOVL2 is a molecular regulator of aging in the retina

Methylation of the regulatory region of the elongation of very-long-chain fatty acids-like 2 (ELOVL2) gene, an enzyme involved in elongation of long-chain polyunsaturated fatty acids, is one of the most robust biomarkers of human age, but the critical question of whether ELOVL2 plays a functional role in molecular aging has not been resolved. Here, we report that Elovl2 regulates age-associated functional and anatomical aging in vivo, focusing on mouse retina, with direct relevance to age-related eye diseases. We show that an age-related decrease in Elovl2 expression is associated with increased DNA methylation of its promoter. Reversal of Elovl2 promoter hypermethylation in vivo through intravitreal injection of 5-Aza-2'-deoxycytidine (5-Aza-dc) leads to increased Elovl2 expression and rescue of age-related decline in visual function. Mice carrying a point mutation C234W that disrupts Elovl2-specific enzymatic activity show electrophysiological characteristics of premature visual decline, as well as early appearance of autofluorescent deposits, well-established markers of aging in the mouse retina. Finally, we find deposits underneath the retinal pigment epithelium in Elovl2 mutant mice, containing components&nbsp;found in&nbsp;human drusen, a pathologic hallmark of age related macular degeneration. These findings indicate that ELOVL2 activity regulates aging in mouse retina, provide a molecular link between polyunsaturated fatty acids elongation and visual function, and suggest novel therapeutic strategies for the treatment of age-related eye diseases

Demographic characteristics by the dichotomized PHQ-8 score (PHQ-8>10) and dichotomized anxiety score (GAD-7 >10) during the pandemic.

Demographic characteristics by the dichotomized PHQ-8 score (PHQ-8>10) and dichotomized anxiety score (GAD-7 >10) during the pandemic.</p