Effect of Peritoneal Fluid from Endometriosis Patients on Sperm Motion Characteristics and Acrosome Reaction

ABSTRACT: Objective-To determine whether peritoneal fluid from women with endometriosis contributes to infertility by impairing sperm motion and functional characteristics. Methods-Women with endometriosis (n = 20) underwent laparoscopy for infertility or pelvic pain. Patients undergoing tubal ligation served as controls (n = 14). Peritoneal fluid was aspirated from women with endometriosis, or from women undergoing laparoscopic tubal ligation. Sperm motility, motion characteristics and acrosome reaction were assessed following incubation with peritoneal fluid. Results-Sperm motility, motion characteristics, and acrosome reaction did not differ significantly between the two groups after 3, 5, or 24 hours of incubation with peritoneal fluid. Conclusions-Sperm motion or functional characteristics showed no significant impairment when sperm from normal donors were incubated with peritoneal fluid from patients with endometriosis. It is unlikely that peritoneal fluid in these patients contributes to infertility. Int J Fertil 44 (1)

Cataract-causing αAG98R-crystallin mutant dissociates into monomers having chaperone activity

Purpose: The G98R mutation in αA-crystallin is associated with autosomal dominant cataract in humans. We have reported that mutant G98R protein has substrate-dependent chaperone activity. Further studies on this G98R mutant protein revealed that mutant protein shows reduced oligomeric stability and accelerated subunit dissociation at a low protein concentration. The purpose of present study was to investigate the chaperone function of dissociated subunits of αAG98R-crystallin. Methods: Substitution of glycine with arginine at position 98 in human αA-crystallin was accomplished by site-directed mutagenesis. The recombinant protein was expressed in E .coli cells and purified by chromatographic techniques. Purified αAG98R-crystallin was diluted to a concentration of 0.1 mg/ml in 50 mM phosphate buffer containing 150 mM NaCl (pH 7.2) and incubated at 37 °C for 24 h. The monomeric subunits were isolated from the oligomers through 50 kDa cutoff filters. The monomers were analyzed by SDS-PAGE, mass spectrometry, and circular dichroism spectroscopy and characterized by multi-angle light-scattering methods. Chaperone activity was tested against four client proteins: citrate synthesis, alcohol dehydrogenate, bovine βB2-crystallin and ovotransferrin. Results: Gel filtration studies showed that αAG98R-crystallin oligomers dissociate readily into monomers. Subunits of αAG98R-crystallin, isolated either by size exclusion chromatography or filtration showed chaperone activity against heatdenatured alcohol dehydrogenase, citrate synthase, bovine βB2-crystallin, and chemically denatured ovatransferrin. SDS-PAGE analysis of the mutant protein incubated at 37 °C for 12 days showed autolysis, which was confirmed by matrixassisted laser desorption ionization time-of-flight mass spectrometry (MALDI TOF MS/MS) analysis of αAG98R-crystallin fragments recovered after SDS-PAGE. Conclusions: The present study shows that the G98R mutation in αA-crystallin produces unstable oligomers that dissociate into active chaperone subunits. The chaperone activity of the dissociated subunits against four client proteins suggests that the αA-crystallin subunits are the minimal units of chaperone activity. α-Crystallin belongs to the family of small heat shock proteins (sHSP) and is composed of two subunits, αA-and αB-, which form heteromers and homomers with varying number of subunits Like other sHSPs, α-crystallin prevents aggregation of denaturing protein

Pharmacological activities and molecular mechanisms of pure and crude extract of Andrographis paniculata: An update

Background: Andrographis paniculata (Burm.f.) Nees (Acanthaceae) is a herbaceous plant used in traditional medicines for the treatment of various ailments. Diterpene lactones are the main phytoconstituents and are responsible for the plant's bitter taste and therapeutic activity. Pure andrographolide (AD) and crude extracts have all been shown to be accountable for various pharmacological activities, including anticancer, anti-inflammatory, hepatoprotective, immunomodulatory, neuroprotective, antidiabetic and antibacterial agents. This review covers recent studies of the exploitation of A. Paniculata with a specific focus on the plant's pharmacological and clinical studies alongwith their therapeutic drug targets and novel drug delivery system. Method: Information for the current review article is collected from various electronic scientific databases such as Science Direct, PubMed, Scopus, Scifinder, Google Scholar, books and reports. All the composed information is classified into different sections according to the objective of the paper. To fetch the useful articles various keywords, namely 'Andrographis paniculata’, ‘King of Bitters’, ‘Andrographolide’ ‘Clinical study’, Molecular mechanism’, ‘Anticancer’, 'Anti-inflammatory', 'Antidiabetic', 'Neuroprotective', ‘Pharmacokinetics’, ‘Novel Drug Delivery System’ were used for search. Results: Of all the 180 studies, twenty one primarily focused on introduction, twenty-seven studies presented a brief introduction of different diseases, ninety studies highlighted the pharmacological activities and therapeutic targets of plant extract and its pure bioactive compounds, twenty five investigations demonstrated the pharmacokinetic parameters of AD and its novel drug delivery systems and seventeen studies investigated clinical studies of the plant. Conclusion: Different recent research and review papers have been studied to gain insights into the therapeutic activities and clinical studies of A. paniculata. Following the extensive literature survey, we collected notable information on the plant. For rapid glance studies, experimental and research findings are compiled into tabular form for quick understanding. Possible directions for future research are also outlined briefly, encouraging the researchers for advance investigations on various mechanistic targets of this plant

Mouse blastocyst previtrification interventions and DNA integrity

Objective: To assess the effect of implementing different previtrification interventions on the postwarming DNA integrity of vitrified blastocysts. Design: Prospective in vitro study. Setting: Center for Reproductive Medicine laboratory in a tertiary hospital setting. Animals: A total of 70 expanded and 46 nonexpanded mouse blastocysts were used for the study

Vitamin C and vitamin E supplementation reduce oxidative stress-induced embryo toxicity and improve the blastocyst development rate. Fertility and Sterility

Objective: To evaluate the adverse effects of exogenously induced reactive oxygen species (ROS) on mouse embryo development by using the 12-phorbol 13-myristate acetate (PMA)-activated leukocyte model as a source of ROS, and to examine the protective effect of antioxidant supplementation (vitamin C and vitamin E). Design: Prospective study. Setting: Research laboratory. Main Outcome Measure(s): Effects of ROS on the blastocyst development rate in the presence and absence of antioxidant supplementation. Result(s): After incubation with the PMA-activated leukocyte supernatant, the median (25th, 75th percentile) rate of blastocyst development significantly decreased from 73% (60%, 80%) after 3 hours to 30% (20%, 40%) after 6 hours compared with control reactions (86% [80%, 100%]). Co-incubating the embryos with vitamin C (50 M) and the PMA-activated supernatant significantly increased the blastocyst development rate from 73% (60%, 80%) to 90% (80%, 91%) at 3 hours and from 30% (20%, 40%) to 91% (89%, 91%) at 6 hours-a level similar to that of control reactions. The blastocyst development rate increased after vitamin E supplementation (400 M) at 6 hours, but not significantly and not by as much as after vitamin C supplementation. Conclusion(s): Exposure of mouse embryos to ROS for extended periods results in embryotoxicity. Vitamin C is more effective than vitamin E in reversing ROS-induced mouse embryo toxicity. (Fertil Steril 2002;78: 1272-7