Impact of embryo co-culture with cumulus cells on pregnancy & implantation rate in patients undergoing in vitro fertilization using donor oocyte.

BACKGROUND & OBJECTIVES: Cumulus cell co-culture of embryo had been found to be beneficial for achieving better pregnancy and implantation rate (IR). The present study was aimed to evaluate efficiency of cumulus co-culture technique over simple culture of embryo in terms of pregnancy rate (PR) and IR in patients undergoing treatment for infertility using donor oocytes fertilized by intracytoplasmic sperm injection. METHODS: This was a quasi-experimental study between control and study groups. The primary endpoint was achievement of pregnancy. Control group included 508 women who underwent embryo development without cumulus cell co-culture and study group included 394 women who underwent embryo development with cumulus cell co-culture using donor's cumulus cells. RESULTS: The present study demonstrated a significant increase in the IR (37.2 vs 24.2%, P<0.001) and in PR (45.7 vs 37.8%, P<0.05) in study group than in control group. The PR and IR were found to be higher in study group, among all groups of women, grouped on the basis of different indications for use of donor oocytes. INTERPRETATION & CONCLUSIONS: Cumulus cell co-culture technique was found to be more effective than simple culture technique for embryo development in women undergoing treatment for infertility using donor oocytes fertilized by intracytoplasmic sperm injection.

Isolation and characterization of nickel uptake by nickel resistant bacterial isolate (NiRBI).

OBJECTIVE: Bioremediation technology has gained importance because microbes could be the convenient source of bio-absorption/bioaccumulation of metals from effluent streams. METHODS: The nickel-resistant bacterial isolates (NiRBI) were selected from various bacterial isolates from industrial effluent and grown in nutrient broth containing different concentrations of nickel sulfate (0.3-3.0 mmol/L) and their capability of accumulating metal from the medium. RESULTS: Well-defined growth of NiRBI was observed in the medium containing up to 2.5 mmol/L of nickel. The isolate was identified using 16S rRNA and closely related to Pseudomonas fragi. Maximum accumulation of nickel (0.59 mg/g dry weight of bacterial cells) was observed when NiRBI was grown in media containing 2 mmol/L of nickel. The protein profile of the NiRBI cellular extract by SDS-PAGE showed two metal stress-induced proteins of molecular weight 48 KD and 18 KD with a simultaneous down regulation of four proteins of 46.7 KD, 42.2 KD, 19.7 KD, and 4.0 KD. CONCLUSION: 48 KD and 18 KD proteins play a role in metal resistance mechanism by NiRBI.

Biodegradable Gelatin Methacrylate Gel as a Potential Scaffold for Bone Tissue Engineering of Canine Adipose-Derived Stem Cells.

Therapeutic potential of adipose derived stem cells (ADSCs) has widely been explored for treatment of orthopedic ailments. Transplantation of cells encapsulated in a scaffold facilitates 3 dimensional modelling of the tissue for the cases where well-defined spatial distribution of cells is desired for implantation. Present study aims to encapsulate canine ADSCs (cADSCs) in biodegradable methacrylated gelatin gel (GelMA) scaffold followed by their osteogenic differentiation for fabrication of a three dimensional bone tissue construct. Different percentages (5, 10 and 20%) and different methacrylation levels of gel (GelMAhigh and GelMAlow) were tested for degradation. Porosity of 10% GelMA was compared by SEM imaging. Gels with the fastest degradation rate (5% GelMAhigh and GelMAlow) were chosen for cell encapsulation since degradation of scaffold is of prime importance when the gel is intended to be used for implantation. Finally, cADSCs encapsulated in 5% GelMAlow demonstrated best morphology and were differentiated osteogenically. We developed a modified protocol for isolation of RNA from cells encapsulated in GelMA. Osteogenic differentiation was affirmed by the presence of osteo-specific gene expression by reverse transcriptase PCR in addition to von Kossa staining of the construct. GelMA is an excellent biodegradable scaffold for encapsulation of cADSCs without altering their osteogenic potential. This osteo-induced cellular scaffold implant opens a new therapeutic horizon in the area of tissue engineering in orthopedics.

Cellular Effect of Curcumin and Citral Combination on Breast Cancer Cells: Induction of Apoptosis and Cell Cycle Arrest.

PURPOSE: The unmanageable side effects caused by current chemotherapy regimens to treat cancer are an unresolved problem. Although many phytonutrients are useful as chemoprevention without side effects, their effects are slower and smaller than conventional chemotherapy. In the present work, we examined the cumulative effect of two phytonutrients, curcumin and citral, on breast cancer cell lines and compared their effect with the known chemotherapy regimen of cyclophosphamide, methotrexate, and 5-fluorouracil. METHODS: Using cultured breast cancer and normal epithelial cells, the cytotoxic and apoptotic effect of curcumin and citral was evaluated in vitro. The synergistic effect of curcumin and citral was calculated by a combination index study using the method by Chou and Talalay. Cell death pathways and mechanisms were analyzed by measuring intracellular reactive oxygen species (ROS) and apoptotic protein levels. RESULTS: Curcumin and citral caused dose and time dependent cell death and showed a synergistic effect at effective concentration EC50 and above concentrations in breast cancer cells without disturbing normal breast epithelial cells. With combination curcumin and citral treatment, apoptosis induction and cell cycle arrest at G0/G1 phase in breast cancer cells were observed. Curcumin and citral generated ROS and activated p53 and poly (ADP-ribose) polymerase-1 mediated apoptotic pathways. CONCLUSION: The results of this study suggest that curcumin and citral in combination may be a useful therapeutic intervention for breast cancer.

Optimization of cellulase production by Penicillium oxalicum using banana agrowaste as a substrate.

The purpose of this study was to produce a higher amount of cellulase by using an alternative carbon source, such as banana agrowaste, and to optimize the fermentation parameters for a high yield. In the present study, cellulase-producing Penicillium was isolated from a decaying wood sample. Different nutritional and environmental factors were investigated to assess their effect on cellulase production. The highest crude enzyme production was observed at a pH 6.0 and a temperature of 28°C in a medium that was supplemented with banana agrowaste as the carbon source. Pretreatment with 2N NaOH, at 7% substrate (banana agrowaste) concentration yielded the highest cellulase activity. Further to this, the effect of other parameters such as inoculum age, inoculum size, static and agitated conditions were also studied. It is concluded that Penicillium oxalicum is a powerful cellulase-producer strain under our tested experimental conditions using banana agrowaste as the carbon source.

Identification of Helicobacter pylori by different conventional staining techniques and its comparison with polymerase chain reaction.

OBJECTIVE: To evaluate Helicobacter pylori (H. pylori) detection by histological staining methods, and to compare with those of Gram staining and polymerase chain reaction (PCR). METHODS: This is a cross-sectional study conducted at the Department of Microbiology, Shree P. M. Patel Paramedical College, Anand, Gujarat, India on 436 patients attending the Deep Surgical Hospital, Anand, Gujarat between February 2008 and October 2011. Biopsies were subjected to histological staining using Hematoxylin & Eosin (H&E), Giemsa, and Warthin-Starry stains, as well as with Gram staining. The PCR was performed on 71 biopsy samples. RESULTS: Sensitivity and negative predictive values of all 3 histological stains (Warthin-Starry, H&E, and Giemsa) were excellent. Gram staining showed excellent results pertaining to sensitivity, specificity, positive predictive value, and accuracy. Sensitivity of PCR was remarkably low compared to all the staining methods. The sensitivity of all histological stains was found better than PCR. CONCLUSION: From the findings in our study, we conclude that in a mediocre laboratory, where PCR facility is not available, histological stain can be a better substitute for the diagnosis of H. pylori. Our findings also confirm the assertion that Gram staining is a preferred stain, affordable, reliable, and simple means for identifying H. pylori compared with both histology and PCR.

Molecular modeling on inhibitor complexes and active-site dynamics of cytochrome P450 C17, a target for prostate cancer therapy.

A molecular model for the P450 enzyme cytochrome P450 C17 (CYP17) is presented based on sequence alignments of multiple template structures and homology modeling. This enzyme plays a central role in the biosynthesis of testosterone and is emerging as a major target in prostate cancer, with the recently developed inhibitor abiraterone currently in advanced clinical trials. The model is described in detail, together with its validation, by providing structural explanations to available site-directed mutagenesis data. The CYP17 molecule in this model is in the form of a triangular prism, with an edge of approximately 55 A and a thickness of approximately 37 A. It is predominantly helical, comprising 13 alpha helices interspersed by six 3(10) helices and 11 beta-sheets. Multinanosecond molecular dynamics simulations in explicit solvent have been carried out, and principal components analysis has been used to reveal the details of dynamics around the active site. Coarse-grained methods have also been used to verify low-frequency motions, which have been correlated with active-site gating. The work also describes the results of docking synthetic inhibitors, including the drug abiraterone and the natural substrate pregnenolone, in the CYP17 active site together with molecular dynamics simulations on the complexes.