Foliar application of silicon, selenium, and zinc nanoparticles can modulate lead and cadmium toxicity in sage (Salvia officinalis L.) plants by optimizing growth and biochemical status.

Different techniques have been used to alleviate metal toxicity in medicinal plants; accordingly, nanoparticles (NPs) have a noticeable interest in modulating oxidative stresses. Therefore, this work aimed to compare the impacts of silicon (Si), selenium (Se), and zinc (Zn) NPs on the growth, physiological status, and essential oil (EO) of sage (Salvia officinalis  L.) treated with foliar application of Si, Se, and Zn NPs upon lead (Pb) and cadmium (Cd) stresses. The results showed that Se, Si, and Zn NPs decreased Pb accumulation by 35, 43, and 40%, and Cd concentration by 29, 39, and 36% in sage leaves. Shoot plant weight showed a noticeable reduction upon Cd (41%) and Pb (35%) stress; however, NPs, particularly Si and Zn improved plant weight under metal toxicity. Metal toxicity diminished relative water content (RWC) and chlorophyll, whereas NPs significantly enhanced these variables. The noticeable raises in malondialdehyde (MDA) and electrolyte leakage (EL) were observed in plants exposed to metal toxicity; however, they were alleviated with foliar application of NPs. The EO content and EO yield of sage plants decreased by the heavy metals but increased by the NPs. Accordingly, Se, Si, and Zn NPS elevated EO yield by 36, 37, and 43%, respectively, compared with non-NPs. The primary EO constituents were 1,8-cineole (9.42-13.41%), α-thujone (27.40-38.73%), ß-thujone (10.11-12.94%), and camphor (11.31-16.45%). This study suggests that NPs, particularly Si and Zn, boosted plant growth by modulating Pb and Cd toxicity, which could be advantageous for cultivating this plant in areas with heavy metal-polluted soils.

Fas cell surface death receptor/Fas ligand genetic variants in gastric cancer patients: A case-control study.

Background & objectives: Various studies have suggested a correlation between Fas cell surface death receptor/Fas ligand (FAS/FASL) variants and multiple types of cancers. The present study aimed to investigate the association between FAS-670A/G and FASL-844C/T and the synergistic effects of both variants on the risk of gastric cancer (GC) in the Kurdish population of west of Iran. Methods: This study was conducted by polymerase chain reaction-restriction fragment length polymorphism technique using MvaI and BsrDI restriction enzymes in 98 GC patients and 103 healthy control individuals. Results: According to the obtained results, a significant association (P=0.008) of FASL polymorphism among GC patients and the control group was detected. Furthermore, no significant differences were found in the FAS polymorphism frequencies between GC patients and the control group. Codominant and dominant models in FASL polymorphism showed significant protective effects against GC [odds ratio (OR)=0.307, 95% confidence interval (CI) (0.134-0.705), P=0.005; OR=0.205, 95% CI (0.058-0.718), P=0.013 and OR=0.295, 95% CI (0.129-0.673), P=0.004 for models of codominant CC vs. CT, codominant CC vs. TT and dominant, respectively]. Furthermore, the presence of both FAS-670G and FASL-844T alleles represented a significant protective effect against GC occurrence [OR=0.420, 95% CI (0.181-0.975), P=0.043]. Interpretation & conclusions: So far, we believe this is the first study, the results of which suggest that FASL gene variation and its synergistic effects with FAS gene could be associated with the risk of GC in the Kurdish population in the west of Iran.

Human placenta-derived mesenchymal stromal cells transfusion in a critically Ill infant diagnosed with Coronavirus Disease 2019 (COVID-19): A case report.

Coronavirus disease 2019 (COVID-19) is still an emergency in many countries. Herein, we report treatment with human placental-derived mesenchymal stromal cells transfusion (hPD-MSCT) in a critically ill infant diagnosed with COVID-19. A 28-day-old male infant with a history of pneumonia was referred to our center with decreased SpO2 (92%) and fever (38.5 °C). Real-time reverse transcription polymerase chain reaction (RT-PCR) and chest computed tomography (CT) confirmed COVID-19 infection. Considering the deteriorating clinical status of the patient despite the routine treatments (SpO2 82%), human placental derived mesenchymal stromal cells (hPD-MSCs) was transfused to him on day 9 and 11 (7 × 106 cells/session). The patient's general condition started to change 3 days after hPD-MSCT and poor feeding and low SpO2 improved day by day. On day 20, the patient was discharged (SpO2 97%) and our one-year follow-up showed a successful response to the treatment with no reported complications. hPD-MSCT may be considered as a possible treatment option in infants/children diagnosed with COVID-19 who fail to respond to conventional therapies. However, required dose, safety, and mechanistic studies are still warranted to further investigate this treatment.

Royal-jelly-based apitherapy can attenuate damages to male reproductive parameter following nicotine administration.

BACKGROUND: Nicotine administration can generate severe oxidative stress and lipid peroxidation. Royal jelly, with its antioxidant properties, acts as a scavenger of reactive oxygen species. This study describes the apitherapy effects of royal jelly on testicular damage following nicotine administration. METHODS: Forty-eight male BALB/c mice were divided into 8 groups (n = 6): saline, 3 different doses of royal jelly (100, 150, and 200 mg/kg body weight (BW) per day), nicotine (1.5 mg/kg), and 3 different groups of Nic + Roy (1.5 mg/kg of Nic + 100, 150, and 200 mg/kg BW per day of royal jelly). Nicotine was administrated intraperitoneally, and royal jelly was prescribed orally for 10 consecutive days. Serum levels of hormones (testosterone, luteinizing hormone, and follicle-stimulating hormone), total antioxidant capacity, nitric oxide (NO) status, malondialdehyde levels, sperm DNA fragmentation, sperm parameters, histopathological changes (H&E staining), immunohistochemistry against apoptotic proteins, and gene expression of Bcl-2, p53, Caspase-3, and Nrf2 (real-time PCR) were assessed to evaluate the molecular and histological changes. RESULTS: Hormone levels, sperm parameters, and status of antioxidants were decreased significantly (p < .05) following nicotine administration. Moreover, royal jelly treatment normalized hormonal and antioxidant characteristics, decreased apoptotic gene expression, increased Nfr2 gene expression, and restored histopathological alteration to the physiological status significantly (p < .05). CONCLUSION: Royal jelly upregulates the antioxidant status, inhibits the mitochondrial-dependent apoptosis pathway, and increases the rate of proliferation. This therapeutic agent effectively protected the testis against nicotine-associated damages by antioxidant and anti-apoptotic effects.

New rat to mouse xenograft transplantation of endometrium as a model of human endometriosis.

Background: Endometriosis can lead to infertility. Since there is no definitive treatment for endometriosis, animal modelling seems necessary to examine the possible treatments. Mouse endometrium cannot be separated for endometriosis induction. In addition, transplantation of uterus into the abdominal viscera to induce endometriosis causes organ damage. In this study, we defined a new model of endometriosis leading to separability of endometrium and a safe anatomical region for transplantation. Methods: Forty female mice were allocated to 5 groups: 1, sham; 2, allograft uterus transplantation of mice to anterior abdominal wall of mice; 3, allograft uterus transplantation of mice to mesentery of mice; 4, xenograft endometrial transplantation of rat to anterior abdominal wall of mice; 5, xenograft endometrial transplantation of rat to mesentery of mice. Adult female rats with a previous pregnancy experience were selected and placed in the vicinity of male rats for 2 weeks to induce estrogen secretion and increase endometrial thickness. Results: In the 4th group of animals, compared to sham, the peritoneal concentrations of VEGF-A, TNF-α, NO, MDA, and serum levels of CA-125 and IL-37 were increased and total body weight was decreased, while weight and size of endometrial lesions were increased significantly (P < .05). Genes expression of HOXA10 and HOXA11 were decreased significantly (P < .05) in groups 2 and 4 compared to sham. Conclusions: Xenograft transplantation of endometrium from rat to anterior abdominal wall of mice can potentially mimic human endometriosis morphologically, histologically, and genetically.

Evaluation of The Relationship among The Levels of SIRT1 and SIRT3 with Oxidative Stress and DNA Fragmentation in Asthenoteratozoospermic Men.

BACKGROUND: Reactive oxygen species (ROS) play a crucial role in etiology of DNA fragmentation and lipid peroxidation in sperm, leading to infertility in men. The silent information regulators SIRT1 and SIRT3 are members of the sirtuins protein family known to be involved in cancer genetics, aging and oxidative stress responses. The aim ofthis study is to determine the correlation between SIRT1 and SIRT3 with antioxidants, oxidative stress biomarkers, and DNA fragmentation in the semen of asthenoteratozoospermic and normozoospermic men. MATERIALS AND METHODS: In this case-control study, after spermogram analysis the specimens were divided into two groups, normozospermic (n=40) and asthenoteratozoospermic (n=40), according to World Health Organization (WHO) standards. Sperm DNA fragmentation was evaluatedusing the sperm chromatin dispersion (SCD) test.Catalase activity was measured using the Aebi spectrophotometeric method. Total antioxidant capacity (TAC) level and superoxide dismutase (SOD) activitywere measured by using commercially available colorimetric assays. Enzyme-linked immune sorbent assay (ELISA) was used to measure SIRT1 and SIRT3 protein levels of seminal plasma. Malondialdehyde (MDA) level in seminal plasma was determined by high-performance liquid chromatography (HPLC). RESULTS: The asthenoteratozoospermic group had significantly lower catalase and SOD activities and TAC levels in comparison with the normozoospermic group (P<0.001).The percentage of DNA fragmentation and MDA level in the asthenoteratozoospermic group were remarkably higher than in the normozoospermic group. The SIRT1 and SIRT3 protein levels in seminal plasmawere remarkably lower in asthenoteratozoospermic group than the normozoospermic group (P<0.001). CONCLUSION: The results of this study suggest that SIRT1 and SIRT3 protein levels are negatively correlated with oxidative stress and DNA fragmentation in semen. The low levels of SIRT1 and SIRT3 in asthenoteratozoospermic men may lead to an increase in oxidative stress, DNA fragmentation, and lipid peroxidation that eventually result in immotile and immature spermatozoa (asthenoteratozoospermia).

Enhancing a Successful Pregnancy and Delivery After ICSI in Advanced-Age Woman with Concurrent Disorders: A Case Report.

BACKGROUND: The objective of this case presentation was describing a live birth in an advanced-age woman with an extremely enlarged uterus, an ovary with blocked fallopian tubes, hypothyroidism and generalized anxiety disorder caused by child-birth following intracytoplasmic sperm injection/embryo transfer (ICSI-ET) with autologous oocytes. CASE PRESENTATION: A 47-year-old patient with an enlarged uterus due to recurrent multiple fibroids following myomectomy was referred to clinical laboratory with a high level of desire to follow the prescribed recommendations and approaches to retrieve her fertility. The patient underwent two cycles of oocyte retrieval and two rounds of frozen-thawed embryo transfer. To achieve a successful pregnancy after oocyte retrieval (birth weight of 3300 g at 38 weeks of gestation), a frozen/thawed embryo in the second cycle of ET was transferred. CONCLUSION: Usage of efficient planning and management of ICSI treatments in patient with autologous oocytes and concurrent disorders, can be used as a new approach to cure the affected individuals.

Autophagy related gene expression status in patients diagnosed with azoospermia: A cross-sectional study.

BACKGROUND: Autophagy affects various aspects of the male reproductive system. Any defects in this process may lead to azoospermia. However, the exact molecular mechanisms of the autophagy pathway have remained largely obscure. Therefore, the present study aimed to investigate levels of autophagy pathway gene expression (i.e. Lc3B, Beclin1, ATG5 and Bcl2) in azoospermic patients. METHODS: The levels of Lc3B, Beclin1, ATG5 and Bcl2 mRNA expression in azoospermic patients and fertile males were evaluated by a real-time polymerase chain reaction technique. In addition, diagnostic evaluation based on the receiver-operating characteristic (ROC) curve was performed. RESULTS: The results obtained showed the decreased expression of Lc3B, Beclin1 and ATG5 genes in infertile patients compared to the control group (p < 0.05), whereas Bcl2 expression was increased in samples (p < 0.05). A diagnostic evaluation by ROC curve and calculation of the area under the curve showed that, using a cut-off relative quantification of 4.550, 0.052, 0.056 and 0.012, the sensitivity of Lc3B, Beclin1, ATG5 and Bcl2 genes was 87.5%, 93.8%, 93.8% and 90%, respectively. In addition, a specificity of 76.7%, 76.7%, 93.3% and 81.2%, respectively, was observed. CONCLUSIONS: As a first study, the current research suggests that an alteration in the expression of autophagy pathway genes may be associated with male infertility. Based on our finding, the increased expression of Bcl2 and formation of Becline1/Bcl2 complex, which inhibits Beclin1 recruitment, may lead to a decrease of the autophagy process in azoospermic patients. Accordingly, upon further investigation, the autophagy could be considered as an important aspect during spermatogenesis.

In vitro effects of clomiphene citrate on sperm parameters and fertilisation rate in male NMRI mice.

Clomiphene citrate (CC), as a medication in male infertility, improves the sperm parameters in oral consumption but various detrimental side effects have been reported including testicular tumours, gynaecomastia, skin allergic reactions and ocular symptoms. Therefore, this study was designed to evaluate the in vitro effects of CC on sperm parameters and fertilisation rate in IVF protocol. Sperm samples of NMRI adult mice were divided into six groups: group 1 received no treatment (control group), while groups of 2, 3, 4, 5 and 6 (experimental groups) were incubated with the doses of 0.001, 0.01, 0.1, 1 and 10 µg/ml of CC in culture medium respectively. Sperm parameters (viability, morphology and motility), DNA fragmentation levels and fertilisation rate in IVF were evaluated. The results demonstrated that the doses of 0.1 µg/ml (p = .000007 for viability and p = .00006 for fertilisation rate) and 1 µg/ml (p = .032 for viability and p = .005 for fertilisation rate) CC cause a significant improvements; also, the dose of 0.1 µg/ml CC found effective on sperm motility (p = .0003). In the field of IVF, the application of 0.1 and 1 µg/ml of CC in the culture medium may improve the sperm parameters in IVF protocol with no side effects.

Protective efficiency of Ashrasi date palm hydroalcoholic extract against diabetes-induced testicular toxicity: A biochemical and stereological study.

The present study was designed to investigate the protective effects of hydroalcoholic extract of Ashrasi date palm (ADP) on diabetes-induced testicular injuries. Adult male rats were randomly allocated into five groups (n = 8 in each group): 1: control; 2: diabetic; 3: diabetic + 30 mg/kg of ADP extract; 4: diabetic + 90 mg/kg of ADP extract; and 5: diabetic + 270 mg/kg of ADP extract. Diabetes was induced by a single dose of streptozotocin (55 mg/kg) intraperitoneally. Testicular changes were assessed quantitatively using stereological method followed by measuring antioxidant enzymes including catalase, superoxide dismutase and glutathione peroxidase and serum testosterone level. Malondialdehyde (MDA) and Bcl-2 expression were also evaluated in tissue samples. Diabetes resulted in significant deleterious alterations in the architecture of testicular tissue, suppressed antioxidant enzymes and testosterone levels and increased lipid peroxidation. The expression of Bcl-2 was downregulated in diabetic testis and resulted in enhanced apoptosis. Eight weeks of ADP extract treatment especially at higher doses could markedly improve structural changes of testis and restore the antioxidant defence and testosterone levels in testicular tissue. In conclusion, this findings showed that ADP extract can play as a potent antioxidant and can attenuate the adverse effects of diabetes on male reproduction.

An Experimental Study to Evaluate the Protective Effects of Solanum lycopersicum Seed Essential Oil on Diabetes-Induced Testicular Injuries.

Background and Objectives: Diabetes is a chronic metabolic disorder that can effectively influences male reproductive performance. The present study was conducted to investigate the protective effects of Solanum lycopersicum essential oil (SL) on diabetes-induced testicular injuries. Materials and Methods: Adult male rats were randomly allocated into five groups (n = 8 in each group). 1: control; 2: diabetic; 3: diabetic + 30 mg/kg of SL essential oil; 4: diabetic + 90 mg/kg of SL essential oil; 5: diabetic + 270 mg/kg of SL essential oil extract. Diabetes was induced by a single dose of streptozotocin (55 mg/kg) intraperitoneally. Testicular changes were assessed quantitatively using stereological method followed by measuring antioxidant enzymes including catalase, superoxide dismutase, and glutathione peroxidase, and the serum testosterone level. Malondialdehyde (MDA) levels and Bcl-2expression were also evaluated in the tissue samples. Results: Diabetes resulted in significant deleterious changes in the structure of testicular tissue, suppressed antioxidant enzymes and testosterone levels, and increased lipid peroxidation. The expression of Bcl-2 was downregulated in diabetic testis and resulted in enhanced apoptosis. Following 8 weeks of treatment with SL essential oil, there were noticeable improvements in the structural changes of testis and the restoration of antioxidant defense and testosterone levels in testicular tissue, especially at higher doses. Conclusion: In conclusion, these findings reveal that the essential oil of Solanum lycopersicum has potent antioxidant properties and can attenuate the adverse effects of diabetes on male reproduction.

Association between the FAS/FASL Variants and Risk of Male Infertility in Asian Populations; A Systematic Review and Meta-Analysis.

Background and Objectives: Studies suggest that FAS/FASL polymorphisms are associated with male infertility; however, their results are still inconclusive. Therefore, this systematic review and meta-analysis aimed to summarize and clarify the overall association of FAS/FASL polymorphisms and risk of male infertility. Materials and Methods: Our search was conducted on the databases of Science Direct, PubMed and Google Scholar. For performing the meta-analysis, pooled odds ratio (OR) values with 95% confidence interval (CI) was applied in order to analyze the strength of association between the FAS/FASL polymorphisms and risk of male infertility. A total of seven relevant studies published up to September 2018 were considered. Results: FASL-844C/T genotype results of 559 patients and 623 healthy individuals were included in our study. For FAS-670A/G genotype effect, 751 patients and 821 healthy individuals were explored. Results showed that all analysis models including dominant, recessive and allelic models of FASL-844C/T and FAS-670A/G polymorphism had no significant effect on infertility in men (p > 0.05 and p > 0.05, respectively). According to sensitivity analysis, our results were stable. Conclusion: We demonstrated that FAS/FASL polymorphisms might not be an effective factor on male reproductive health. For precise determination of FAS/FASL polymorphisms effects on male infertility, large-scale case-control studies should be performed.

Association of FAS-670A/G and FASL-844C/T polymorphisms with idiopathic azoospermia in Western Iran.

OBJECTIVE: The FAS/FASL interaction plays a central role in up-regulation of apoptosis in testis. Studies indicated that the FAS-670A/G and FASL-844C/T polymorphisms are associated with the risk of idiopathic azoospermia in different ethnic groups. Therefore, the current study aims to investigate the association between FAS-670A/G and FASL-844C/T polymorphisms with male idiopathic infertility in Western Iran. STUDY DESIGN: The analysis of FAS-670A/G and FASL-844C/T polymorphisms were carried out using the PCR-RFLP approach, on 102 infertile men and 110 normal fertile men as control group. RESULTS: The results suggested that there were no significant difference in genotypic frequencies of FAS-670A/G polymorphism between infertile and control groups. On the other hand, significant result was observed for the frequency of FASL-844C/T polymorphism in infertile men in comparison to control group (P=0.02). Indeed, men with FASL-844TT and CT genotypes had an increased risk of idiopathic azoospermia in comparison to those with CC genotype (OR=2.02, 95% CI [1.05-3.88, P=0.03] and OR=1.44, 95% CI [0.46-4.49, P=0.53]), respectively. CONCLUSION: Our findings speculate that the FASL-844C/T polymorphism is associated with the risk of male infertility and this variation can be considered as a genetic risk factor for idiopathic azoospermia among Western Iranian men population. Summing up, these data indicated that the genetic variations in FAS/FASL system have a critical role in spermatogenesis defects and subsequent male infertility.

New procedure for epidermal cell isolation using kiwi fruit actinidin, and improved culture of melanocytes in the presence of leukaemia inhibitory factor and forskolin.

OBJECTIVES: Conventional isolation of epidermis from the dermis and disruption of epidermal sheets to liberate the cells, are performed using proteolytic enzymes such as thermolysin or collagenase. Selective population expansion of melanocytes is achieved by suppressing proliferation of keratinocytes and fibroblasts in epidermal cell suspensions, using phorbol esters and cholera toxin. Here, we introduce a new procedure for isolation of epidermal cells, using proteolytic activity of kiwi fruit actinidin, and also an improved growth medium for melanocytes in the presence of leukaemia inhibitory factor (LIF) and forskolin. MATERIALS AND METHODS: Dermo-epidermal separation and epidermal sheet cell dispersion were performed using actinidin compared to conventional proteases including collagenase, thermolysin or trypsin. Thereafter, melanocyte culture was performed in two common media and one modified medium to discover optimization for these cells. RESULTS: We found that dermo-epidermal separation and epidermal sheet cell dispersion using kiwi fruit actinidin were considerably better than previously used methods, both from the aspect of less fibroblast and keratinocyte contamination, and of more viable native cells. Also, melanocytes proliferated better in phorbol ester- and cholera toxin-free proliferation medium supplemented with LIF and forskolin. CONCLUSION: Less contamination and higher numbers of viable cells were actinidin preferential for separation of epidermis and isolation of epidermal cells. Supplementation of LIF and forskolin to new medium increased proliferation potential of melanocytes in comparison to exogenous mitogens.