Sperm DNA fragmentation negatively influences the cumulative live birth rate in the intracytoplasmic sperm injection cycles of couples with unexplained infertility.

OBJECTIVE: This study aimed to determine the effect of sperm DNA fragmentation (SDF) on the cumulative live birth rate (CLBR) in intracytoplasmic sperm injection (ICSI) cycles in couples with unexplained infertility. METHODS: We conducted a prospective study of 145 couples who underwent ICSI cycles for unexplained infertility. Based on the SDF rate, patients were categorized into a low SDF group (SDF ≤30%, n=97) and a high SDF group (SDF >30%, n=48). SDF was assessed using the acridine orange test on density gradient centrifugation prepared samples. The CLBR was calculated as the first live birth event per woman per egg collection over 2 years. RESULTS: The high SDF group (SDF >30%) showed a significantly lower CLBR (p<0.05) and a significantly higher miscarriage rate (p<0.05) than the low SDF group (SDF ≤30%). No significant difference was observed in the implantation and cumulative pregnancy rates between the two SDF groups. The total number of embryo transfers was stratified further into fresh and frozen embryo transfers. In the fresh embryo transfers, there were significant differences in the implantation rates, clinical pregnancy rates, and live birth rates (p<0.05) between the low SDF and high SDF groups. However, in the frozen embryo transfers, there were no significant differences in clinical outcomes between the two groups. In the multivariable logistic regression analysis, SDF was a predictor of CLBR (p<0.05) when adjusted for possible confounding factors. CONCLUSION: High SDF was associated with a lower CLBR and a higher miscarriage rate in the ICSI cycles of couples with unexplained infertility.

Role of Female Age in Regulating the Effect of Sperm DNA Fragmentation on the Live Birth Rates in Intracytoplasmic Sperm Injection Cycles with Own and Donor Oocytes.

Background: Sperm DNA integrity assessment has been progressively used as an unfettered measure of sperm as it proffers more prognostic and diagnostic information than routine semen analysis. The contentious effect of sperm DNA fragmentation (SDF) on clinical outcomes can be attributed to female factors such as age, oocyte quality and ovarian reserve. Aims: The study is mainly aimed to know the influence of SDF on the live birth rates in intracytoplasmic sperm injection (ICSI) cycles with own and donor oocytes. Second, to know the role of female age in regulating the effect of SDF on the live birth rates in ICSI cycles with own and donor oocytes. Setting and Design: A prospective cohort study was done at our tertiary care centre attached to the reproductive medicine unit in medical college. Materials and Methods: The study included 356 patients who underwent first ICSI cycles either with own or donor-oocytes along with day 5 fresh embryo transfers only. The main outcome measures were live birth rates and miscarriage rates. Statistical Analysis Used: Chi-squared test was used to compare the categorical variables between the groups. The receiver operating characteristic curve was developed to correlate the female age with the live birth rate. Results: A significant decrease in the live birth rates (42.85% vs. 26.15%, P = 0.023) and an increase in the miscarriage rates (12.30% vs. 34.61%, P = 0.013) were observed in the high-SDF group ICSI cycles of own-oocyte patients. However, there was no significant difference in the live birth rates and miscarriage rates in the low- and high-SDF groups of donor oocyte ICSI cycle patients (P > 0.05). The own-oocyte ICSI cycle patients were further stratified based on the female age. In the female age group ≤30 years there was no significant difference in the live birth and miscarriage rates (P > 0.05) similar to donor oocyte ICSI cycles. Whereas, there was a significant difference in the live birth rates in the females of age >30 years (13.79% vs. 34.37%, P = 0.040). Conclusion: In conclusion, high-SDF has a negative influence on the live birth rates and a positive influence on the miscarriage rates in patients with own-oocyte ICSI cycles. A similar influence was not observed in patients with donor-oocyte ICSI cycles and in young female patients (age ≤30 years) with own-oocyte ICSI cycles.

Phytochemical Profiling of Methanolic Fruit Extract of Gardenia latifolia Ait. by LC-MS/MS Analysis and Evaluation of Its Antioxidant and Antimicrobial Activity.

Gardenia latifolia Ait. (Rubiaceae) is also known as Indian Boxwood is a small deciduous tree often growing in southern states of India. In the present study, phytochemical profiling of methanolic extract of G. latifolia fruits were carried out using FTIR and LC-MS/MS analysis. Besides, its antioxidant and antimicrobial potential have been analysed using DPPH activity, differential pulse voltammetry and resazurin microtiter assay, respectively. Phytochemical profiling revealed the presence of 22 major diversified compounds and main were 3-caffeoyl quinic acid (chlorogenic acid), 3,4-Di-O-caffeoyl quinic acid, 6-O-trans-feruloylgenipin gentiobioside, 10-(6-O-trans sinapoyl glucopyranosyl) gardendiol, isoquercitrin, scortechinones, secaubryenol, iridoids and quercetin 3-rutinoside (rutin). The extract showed antioxidant activity (IC50 = 65.82) and powerful antibacterial activity with lowest minimum inhibitory concentration against Gram-positive Staphylococcus aureus (15.62 µg/µL), Bacillus subtilis (31.25 µg/µL) than gram negative Escherichia coli (62.5 µg/µL), Klebsiella pneumoniae (62.5 µg/µL), Pseudomonas aeruginosa (31.25 µg/µL). This study shows that the fruits of G. latifolia have tremendous potential to be used in food industries, phyto-therapeutics and cosmetic industries.

Biogenesis of silver nanoparticles using leaf extract of Indigofera hirsuta L. and their potential biomedical applications (3-in-1 system).

The present study reports the biosynthesis of silver nanoparticles (IH-AgNPs) using aqueous leaf extract of Indigofera hisruta L. The biosynthesized IH-AgNPs were found to be FCC crystals, 5-10 nm in size, spherical in shape and stable. The biosynthesized IH-AgNPs showed dose-dependant cytotoxicity against prostate cancer (PC3) (IC50 = 68.5 µg/mL), colon cancer (COLO205) (IC50 = 85.2 µg/mL), and mouse melanoma (B16F10) (IC50 = 80.9 µg/mL). IH-AgNPs were found to be nontoxic towards normal CHO (Chinese hamster ovary) cells. The biosynthesized IH-AgNPs showed effective in vitro antioxidant activity against DPPH (IC50 = 63.43 µg/mL) and H2O2 (IC50 = 89.93 µg/mL) radicals. IH-AgNPs exhibited effective antibacterial activity against both Gram+ve and Gram-ve bacteria. MIC values of IH-AgNPs against S. aureus, B. subtilis, P. aeruginosa and E. coli were found to be 7.8 µg/mL, 3.9 µg/mL, 15.6 µg/mL and 15.6 µg/mL respectively. IH-AgNPs also showed inhibitory activity against fungal pathogens including C. albicans, C. nonalbicans and C. tropicalis. Considering the results together, we demonstrate that IH-AgNPs exhibits three different bioactivities (3-in-1 system) and they could be employed as future antimicrobial, antioxidant and anticancer agents/drug delivery vehicles in the field of biomedicine.

Effects of the culture media optimization on pectinase production by Enterobacter sp. PSTB-1.

In the present study, media composition for high production of pectinase by Enterobacter sp. PSTB-1 in submerged fermentation was optimized using response surface methodology (RSM). Mango fruit processing industrial waste (MIW) was used as substrate (carbon source) as it contains high amount of pectin. Enterobacter sp. PSTB-1 used in present study has given pectin clear zone (PCZ) of 34 mm is higher than other isolates. The experimental results made by statistical design for high pectinase production revealed that the suitable media components: NaNO3 2.0 g/l, KCl 0.50 g/l, KH2PO4 1.0 g/l, MgSO4·7H2O 0.50 g/l, Yeast extract 1.0 g/l, mango industrial waste powder 5.0 g/l. The actual pectinase activity was 75.23 % correlated with the predicted pectinase activity where the model (CCD) was significant. Response surface modelling applied effectively to optimize the production of pectinase in submerged fermentation to make the process low cost-effective by using powdered mango industrial waste as substrate.

Bio-catalysis of mango industrial waste by newly isolated Fusarium sp. (PSTF1) for pectinase production.

The dried mango fruit processing industrial waste (MIW) used as carbon source for the production of pectinase from fungal strains. Eight fungal strains were isolated from MIW and screened for their ability to produce pectinase by pectin clear zone (PCZ) technique. Fusarium sp. (PSTF1) showed highest PCZ value of 52 mm. The physico-chemical characteristics of the medium were standardized for high production of pectinase. The highest production of pectinase in submerged fermentation observed at temperature-28 °C, pH-6.0, inoculum-size 0.6/25 ml, incubation-72 h, substrate concentration-0.6 g/100 ml, carbon source-fructose (1 %). The effect of different amino acids, vitamins also observed. Under these optimal conditions the highest activity 81.9718 µmol/ml of pectinase was observed. The Fusarium sp. (PSTF1) has been considered as the best pectinase producer in submerged fermentation of MIW. The cheap waste raw material used as best carbon source for high production of high value pectinase.