Dexamethasone effects on Bax expression in the mouse testicular germ cells.

Exposure to glucocorticoids (GCs) leads to numerous changes in various biological systems including the reproductive system. The aim of the present study was to find out whether dexamethasone (Dex), a widely used GC, would influence the apoptosis and expression of Bax, an important proapoptotic protein, in the mouse testicular germ cells. Experimental groups of 8 male NMRI mice received one of the following treatments daily for 7 days: 4, 7 and 10 mg/kg Dex. Control groups were treated with equivalent volumes of saline. Experimental and control animals were sacrificed 24 h after the last injection. Immunohistochemical procedure was used to evaluation of Bax expression and the deoxyuridine nick-end labeling (TUNEL) was applied to assessment of the apoptotic germ cells. Bax expression was upregulated mainly at stages VII-VIII of spermatogenic cycle (

EXPRESSION AND FUNCTION OF INTEGRIN ASSOCIATED PROTEIN (IAP)/CD47 IN HUMAN ARTICULAR CARTILAGE

ABSTRACT Background: Integrin-associated protein (IAP) /CD47, a member of immunoglobulin super family (IgSF), is expressed in a variety of cell types. In the present work, the expression pattern of IAP/ CD47 in normal and osteoarthritis (OA) human articular cartilage and its role in mechanotransduction pathway following mechanical stimulation have been studied. Methods: Using immunohistochemistry and standard western blotting the in vivo and in vitro expression pattern of CD47 in normal and OA human articular cartilage was assessed.The role of CD47 in mechanotransduction was evaluated by using an electrophysilogical method. Results: Immuno histochemical studies showed a similar very strong expression pattern of CD47 in both normal and OA cartilage. Electrophysilogical experiments showed a role for CD47 in chondrocyte response to mechanical stimulation. Conclusion: Very strong expression pattern of CD47 in both normal and OA chondrocyte may be a result of its critical function/s in chondrocyte homeostasis and it appears that CD47 is important in regulation of the chondrocyte response to mechanical stimulation

Immunohistochemical assessment of galectin-3 during pre-implantation in mouse endometrium

Background: Galectin-3 (Gal-3), a β-galactoside-binding lectin, is a multifunctional lectin that involves in a number of critical biological processes. Objective: The purpose of this study was to investigate the expression pattern of Gal-3 in mouse endometrium during estrus phase of estrous cycle and pre-implantation. Materials and Methods: In this experimental study 42 NMRI female mice were divided in seven different groups. Ovulation in NMRI female mice was stimulated by injecting hMG and hCG. Estrus phase was considered as stimulated and un-stimulated groups. The other groups of mice were mated, and the day of vaginal plug formation was considered as the day 1 of pregnancy. The mice of all groups were sacrificed on different days of pre-implantation period and their uterine horns were fixed and avidin- biotin complex method of immunohistochemistry (IHC) was applied. Results: In estrus group, Gal-3 immunoreactivity in luminal epithelium was strong, in stromal cells very strong, in glandular epithelium very weak and endothelial cells very strong. No identifiable difference was observed in un-stimulated and stimulated estrus phase. In test groups, days 1-2, insignificant difference of Gal-3 expression was observed. On day 3, luminal epithelium and stromal cells showed significant decrease in comparison to estrus and day 1 (p=0.001). On the 4th and 5th days, luminal epithelium and stromal cells showed significant decrease in comparison to estrus phase and days 1-3 (p=0.0001). Conclusion: The data suggested that successful implantation is probably associated with the downregulation of Gal-3 in the mouse endometrium at the beginning of pregnancy

EFFECT OF KIDNEY STEM CELLS ON BIOCHEMICAL ANALAYSIS AND SMAD2/3 PHOSPHORYLATION IN RATS WITH DIABETIC NEPHROPATHY

Introduction: Mesenchymal stem cells (MSCs) were proposed as a critical therapeutic candidate in diabetic nephropathy (DN). Renal stem cells as a source for repairing are controversial. The purpose of the present study was to evaluate the effect of kidney rat stem cells on DN. Materials and methods: After separation of renal stem cells from rat kidney, the surface Stem cell markers were assessed by flow cytometry analysis. To establish the diabetic nephropathy rat model STZ(60mg/kg) was used. The cells were injected to experimental groups via tail vein (2×106cells/rat). Biochemical and histological parameters were evaluated in order to determining the impact of stem cells on kidney structure. Phosphorylation of Smad2/3 two weeks after induction of early diabetic nephropathy was evaluated by using standard western blotting.Result: The cells significantly reduced blood nitrogen (BUN), serum creatinine (Scr) and 24 urinary proteins. The phosphorylation of smad2 and smad3 significantly down-regulated. PAS staining showed in the presence of adult kidney stem cells histopathological changes were improved.Conclusion: Adult kidney stem cells may be a candidate for treatment of early DN to improve the kidney function and regenerating kidney tissues in DN rats

Isolation and differentiation of mouse embryonic stem cells

Background: Recently, embryonic stem (ES) cells have become very important resources in basic medical researches. These cells can differetiate into derivatives of all primary germ layers. Objectives: In order to isolate embryonic stem cells in vitro, the blastocyst were cultured and the morphological aspects, population doubling time, alkalin phosphatse and differentiation properties of the cells were investigated. Materials and Methods: The balstocysts from NMRI mice were cultured for 3 days up to time that inner cell mass (ICM) reach to the outgrowth stage. The cells were disaggregated and trypsinized every 3 days until the appearance of the colonies of ES cells. The colony positive cells were fixed and stained for alkaline phosphatase. The ES cells were cultured in suspension state for 5 days, at the same time Leukaemia Inhibitory Factor (LIF) was removed from media to form embryoid bodies(EBs). The EBs were cultured for 8 - 20 days on collagen coated dish to induce the spontaneouse differentiation. Results: During the 6-9 days after the disaggregation of ICM in the expansion stage, the colony of ES cells appeared as a flat monolayer mass with strike boundaries and nondistinguish cytoplasm including a few nuclei. In colony formation stage, the morphology changed from flat monolayer to round multilayer with strike define boundaries. Undifferentiated cells were seen as intensely small cells attached together compactly with high nucleus/cytoplasm (N/C) ratio. The cells of colonies tend to differetiate by separation from each other and became larger and diffused on substrate by attaching to dish. The positive alkaline phosphatase cells were seen in typical morphology of ES colonies. The EBs cells were seen in culture after 5 days in suspension and began to spontaneously differentiate into various types of cells such as nerve and hematopoitic lineages. Conclusion: Despite strike morphology of ES colonies, it is difficult to distinguish the differentiated from undifferentiated cell colonies in the colony formation stage. New ES cells are capable to give rise into EBs and are susceptible of spontaneously differentiation in various type of cells

Improvement of mesenchymal stem cell differentiation into the endoderm lineage by four step sequential method in biocompatible biomaterial

Introduction: The goal of the study described here, was to investigate the potential of umbilical cord derived mesenchymal stem cell (UC-MSCs) into hepatocyte like cells in a sequential 2D and 3D differentiation protocols as alternative therapy. Methods: Mesenchymal stem cells (MSCs) were isolated from the umbilical cord (UC) and CD markers were analyzed by flow cytometry. For hepatic differentiation of UC-MSCs, cells were induced with a sequential 4-step protocol in 3D and 2D culture system. Urea concentration and albumin secretion into the culture medium was quantified by ELISA. Gene expression levels of AFP, ALB, and CK18 were determined by RT-PCR. Data were statistically analyzed by the SPSS software. The difference between the mean was considered significant when p < 0.05. Results: Growth factor dependent morphological changes from elongated fibroblast-like cells to round epithelial cell morphology were observed in 2D culture. Cell proliferation analysis showed round-shaped morphology with clear cytoplasm and nucleus on the alginate scaffold in 3D culture. The mean valuses of albumin production and urea secretion were significantly higher in the 3D Culture system when compared with the 2D culture (p = 0.005 vs p = 0.001), respectively. Treatment of cells with TSA in the final step of differentiation induced an increased expression of CK18 and a decreased expression of αFP in both the 3D and 2D cultures (p = 0.026), but led to a decreased albumin gene expression, and an increased expression in the 2D culture (p = 0.001). Conclusion: Findings of the present study indicated that sequential exposure of UC-MSCs with growth factors in 3D culture improves hepatic differentiation

Curcumin Attenuates Hepatotoxicity Induced by Zinc Oxide Nanoparticles in Rats

Background: Zinc oxide nanoparticles (NZnO) are increasingly used in modern life. Most metal nanoparticles have adverse effects on the liver. Aims: To explore the protective action of curcumin (Cur) against hepatotoxicity induced by NZnO in rats. Study Design: Animal experimentation. Methods: Control group animals received normal saline, while the Cur group animals were treated with 200 mg/kg of Cur orally for 21 days. NZnO-intoxicated rats received 50 mg/kg of NZnO for 14 days by gavage method. In the NZnO+Cur group, rats were pretreated with Cur for 7 days before NZnO administration. Plasma activities of Alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) were measured as biomarkers of hepatotoxicity. Hepatic levels of malondialdehyde (MDA) and superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities were measured for detection of oxidative stress in liver tissue. Histological changes and apoptosis in liver tissue were studied by using Hematoxylin-eosin staining and the transferase dUTP nick end labeling (TUNEL) method. Results: NZnO induced a significant increase in plasma AST (2.8-fold), ALT (2.7-fold) and ALP (1.97-fold) activity in comparison to the control group (p<0.01). NZnO increased MDA content and reduced SOD and GPx activities. NZnO caused liver damage including centrilobular necrosis and microvesicular steatosis. The percentage of apoptosis in hepatocytes was increased in NZnO-treated rats (p<0.01). Pre-treatment of Cur significantly reduced lipid peroxidation (39%), increased SOD (156%) and GPx (26%) activities, and attenuated ALT (47%), AST (41%) and ALP (30%) activities. Pre-treatment with Cur also decreased the histology changes and apoptotic index of hepatocytes (p<0.05). Conclusion: These findings indicate that Cur effectively protects against NZnO-induced hepatotoxicity in rats. However, future studies are required to propose Cur as a potential protective agent against hepatotoxicity induced by metal nanoparticles

A Proteomic Analysis of Human Follicular Fluid: Comparison between Younger and Older Women with Normal FSH Levels

The follicular fluid (FF) is produced during folliculogenesis and contains a variety of proteins that play important roles in follicle development and oocyte maturation. Age-related infertility is usually considered as a problem that can be solved by assisted reproduction technology. Therefore, the identification of novel biomarkers that are linked to reproductive aging is the subject of this study. FF was obtained from healthy younger (20–32 years old) and older (38–42 years old) women undergoing intracytoplasmic sperm injection (ICSI) due to male factor infertility. The FF was analyzed by two-dimensional gel electrophoresis (2-DE). The power of two-dimensional gel electrophoresis and the identification of proteins were exploited using matrix-assisted laser desorption-ionization time-of-flight/time-of-flight (MALDI-TOF-TOF) mass spectrometry. Twenty three protein spots showed reproducible and significant changes in the aged compared to the young group. Of these, 19 protein spots could be identified using MALDI-TOF-TOF-MS. As a result of MASCOT search, five unique downregulated proteins were identified in the older group. These were identified as serotransferrin, hemopexin precursor, complement C3, C4 and kininogen. A number of protein markers were found that may help develop diagnostic methods of infertility