Cytoprotective and anti-apoptotic effects of Satureja khuzestanica essential oil against busulfan-mediated sperm damage and seminiferous tubules destruction in adult male mice.

We studied the protective effect of Satureja khuzestanica essential oil (SKEO) against damage caused by busulfan on testis in male mice. The NMRI mice (n = 40) were assigned to four groups including: G1: control, G2: treated with busulfan for 4 days (3.2 mg kg(-1)), G3: receive busulfan (4 days, 3.2 mg kg(-1)) and SKEO (28 days, 225 mg kg(-1)) at the same time, G4: pre-treated with SKEO (7 days, 225 mg kg(-1)) and subsequently cotreated with busulfan (4 days, 3.2 mg kg(-1)) and SKEO (28 days, 225 mg kg(-1)). The histological changes of testis were analysed using H&E staining. Sperm parameters, cytotoxic and apoptotic factors were also studied by computer-aided sperm analyzer, MTT and TUNEL assays respectively. Our results showed that SKEO pre-administration significantly improved all parameters of epididymal spermatozoa and decreased germinal epithelium destruction following busulfan chemotherapy. We also found lower MTT levels and TUNEL-positive cells in SKEO pre-treated groups. In conclusion, SKEO possesses beneficial effects on sperm parameters when taken before chemotherapy and continued during and after chemotherapy for a long time, than when used short-term coinciding with the chemotherapy. Our results support valuable data about the application of SKEO for protection against adverse effects of busulfan on male genital system in patients under chemotherapy.

Short-term in-vitro culture of goat enriched spermatogonial stem cells using different serum concentrations.

PURPOSE: To investigate the effect of serum supplementing on short-term culture, fate determination and gene expression of goat spermatogonial stem cells (SSCs). METHODS: Crude testicular cells were plated over Datura-Stramonium Agglutinin (DSA) for 1 h, and non-adhering cells were cultured in the presence of different serum concentrations (1, 5, 10, and 15%) for 7 days in a highly enriched medium initially developed in mice. Colonies developed in each group were used for the assessment of morphology, immunocytochemistry, and gene expression. RESULTS: Brief incubation of testicular cells with DSA resulted in a significant increase in the number of cells that expressed the germ cell marker (VASA). The expression of THY1, a specific marker of undifferentiated spermatogonia, was significantly higher in colonies developed in the presence of 1% rather than 5, 10 and 15% serum. CONCLUSION: Goat SSCs could proliferate and maintain in SSC culture media for 1 week at serum concentrations as low as 1%, while higher concentrations had detrimental effects on SSC culture/expansion.

Embryonic stem cell sphere: a controlled method for production of mouse embryonic stem cell aggregates for differentiation.

OBJECTIVES: Embryonic stem cells (ESCs) are of significant interest as a renewable source of nonproliferating cells. Differentiation of ESCs is initiated by the formation of embryoid bodies (EBs). Standard methods of EB formation are limited in their production capacity, in any variations in EB size and formation of EBs through frequent passages. Here we have reported the utility of a microencapsulation technique for overcoming these limitations by mass production of mouse ESCs in alginate beads called ESC spheres. METHODS: The mouse ESCs were encapsulated in 1.2% alginate solution and cocultured on a feeder layer. The cells were evaluated by flow cytometry, in vitro differentiation, immunofluorescence, and reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: Analysis of encapsulated ESC spheres by flow cytometry showed similar percentages of Oct-4 and stage-specific embryonic antigen-1 (SSEA-1) expression in comparison with routine culture of ESCs. Moreover, the ESC spheres maintained a pluripotency potential which was comparable with ESCs cultured on feeder cells directly, as demonstrated by immunofluorescence and RT-PCR. CONCLUSIONS: The results demonstrated that alginate encapsulation as a simple bioreactor, provides a scalable system for mass undifferentiated ESC sphere production with similar sizes and without the need for frequent passages for differentiation and clinical and pharmaceutical applications.

Interleukin-28B rs12979860 C/T Polymorphism and Acute Cellular Rejection after Liver Transplantation.

BACKGROUND: Interleukin-28 (IL-28B) rs12979860 C/T polymorphism is a known predictor of sustained virological response after antiviral treatment in hepatitis C. IL-28B affects the innate immune system as well as intrahepatic expression level of interferon-stimulated genes. OBJECTIVE: To investigate the effect of recipient IL-28B polymorphism on occurrence of acute rejection after liver transplantation. METHODS: 140 liver allograft recipients were selected. Acute rejection episodes were recorded in 39 patients (AR group); the remaining had normal graft function (non-AR group). 70 normal subjects were also studied as the control group. The IL-28B rs12979860 was genotyped through PCR-RFLP method. RESULTS: No significant difference was found between AR and non-AR groups in terms of genotype and allele frequency. However, the CC genotype was significantly (p<0.001) more frequent in patients than in the control group; the C allele variants increased the risk of end-stage liver disease (OR: 2.60). CONCLUSION: Liver damage in association with the carriage of IL-28B C allele is associated with a higher likelihood of developing cirrhosis.

Generation of Islet-like Cell Aggregates from Human Adipose Tissue-derived Stem Cells by Lentiviral Overexpression of PDX-1.

BACKGROUND: Pancreatic duodenal homeobox1 (PDX-1) is a transcription factor that is important in regulating pancreas development and maintaining ß-cell function. ß-cell replacement is an effective approach for the treatment of type 1 diabetes. Human adipose-mesenchymal stem cells (hAMSCs) are the ideal population cells for differentiating into insulin-producing cells. OBJECTIVE: To determine if islet-like cell aggregates production could be generated from hAMSCs by lentiviral overexpression of PDX-1. METHODS: After isolation of hAMSCs, characteristics of these cells were identified by flow-cytometic analysis and multilineage differentiation studies. PDX-1 gene delivered into hAMSCs through lentiviral vector for differentiating hAMSCs into insulin-producing cells (IPCs) at the utilized protocol for 14 days. Characteristics of IPCs were evaluated by immunocytofluorescence, dithizone staining, and quantitative reverse transcription PCR. In response to high glucose medium, insulin release was detected by chemiluminescence enzyme immunoassay. RESULTS: The islet-like cell aggregates appeared about 10 days after introduction of PDX-1 into hAMSCs. PDX-1 induced its own expression (auto-induction), a number of islet-related genes such as Ngn3, Nkx2-2, and insulin. The insulin-positive cells were detected in the PDX-1 transduced cells. In response to glucose challenge test, secretion of insulin hormone in the medium with high glucose concentration significantly increased in the PDX-1-transduced cells related to medium with low glucose concentration. CONCLUSION: Introduction of lentiviral PDX-1 significantly induces hAMSCs to differentiate into islet-like cell aggregates, which may provide a source of adipose stem cells-derived insulin-producing cells for cell replacement therapy in type 1 diabetes.

Role of subject-specific musculoskeletal loading on the prediction of bone density distribution in the proximal femur.

The typical bone density patterns in the proximal femur can be explained using bone remodeling simulations incorporating a load-adaptive response. Yet, subject-specific variations in bone density have not received much attention. Therefore, the objective of this study was to quantify to what extent subject-specific bone geometry and subject-specific musculoskeletal loading affect the predicted bone density distribution. To accomplish this goal, a computational bone remodeling scheme was combined with gait analysis and a subject-specific musculoskeletal model. Finite element models incorporating the subject-specific geometry as well as the subject-specific hip contact forces and associated muscle forces were used to predict the density distribution in the proximal femur of three individuals. Next, the subject-specific musculoskeletal loads were interchanged between the subjects and the resulting changes in bone remodeling of the proximal femur were analyzed. Simulations results were compared to computed tomography (CT) image-based density profiles. The results confirm that the predicted bone density distribution in the proximal femur is drastically influenced by the inclusion of subject-specific loading, i.e. hip contact forces and muscle forces calculated based on gait analysis data and musculoskeletal modeling. This factor dominated the effect of individualized geometry. We conclude that when predicting femoral density distribution in patients, the effect of subject-specific differences in loading conditions of the hip joint and the associated difference in muscle forces needs to be accounted for.

Evaluation of sperm selection procedure based on hyaluronic acid binding ability on ICSI outcome.

PURPOSE: To compare the efficiency of routine sperm selection method with HA-selection procedure for fertilization rate, embryo development, implantation and pregnancy rates as well as evaluating the relationship between HA-binding ability with sperm protamine deficiency and DNA fragmentation. METHODS: Semen samples were obtained from the 50 couples undergoing ICSI. The percentage of fertilization rate, cleavage and quality of embryos compared between two procedures (routine sperm selection and HA-binding selection). The semen samples were assessed for DNA fragmentation and protamine deficiency by sperm chromatin dispersion (SCD) test and Chromomycin A3 (CMA3) staining, respectively. RESULTS: A significant inverse correlation was observed between percentage of HA binding with protamine deficiency, DNA fragmentation and abnormal sperm morphology (P < 0.05). Furthermore, in current study, oocytes inseminated by HA sperm selection procedure had significantly higher fertilization rate (P < 0.05). While the pregnancy and implantation rates were insignificantly increased. CONCLUSION: The results suggest that normal sperm have higher chance to bind HA and therefore, HA sperm selection procedure may select sperm with normal protamine content and low DNA fragmentation, but to confirm the effect of HA sperm selection on the ICSI outcome requires further studies.