Comparison of conditioned medium and direct co-culture of human granulosa cells on mouse embryo development

Although numerous investigations have demonstrated the beneficial effects of co-culture system of different somatic cells on in vitro development of embryos, the effects of conditioned-media of co-culture cells have not been well documented. The objective of this study was to compare the effects of human granulosa cells co-culture system and its conditioned medium on the developmental rate of mouse embryos in vitro. Two sets of experiments were undertaken: in the first one 317 mouse one-cell embryos were cultured in human granulosa cell co-culture system (GC). Ham's F10 medium conditioned with granulosa cells (CM) and non-conditioned Ham's F10 for 120 h. In the second experiment. 391 late two-cell embryos were cultured in the 3 fore-mentioned culture treatments for 72 h. Embryos were obtained from NMRI mice. Granulosa cells were collected from patients undergoing an IVF program during oocyte pickup. In the first set of experiments, 23.6, 14.5 and 11.1% of one-cell embryos passed two-cell block and continued growing to 4-cell in GC, CM and HF, respectively. This index in GC was significantly different from two other treatments. Also significantly more embryos reached blastocyst stage in GC compared with two other treatments. The blastocyst rate was not significantly different between CM and HF. In the second set of experiments the proportion of blastocyst stage was significantly higher in CM than that in HF and lower than that in GC. In conclusion, although human granulosa cell-conditioned medium has beneficial effects on mouse embryo development, it was not as effective as co-culture of these cells.     

Soils selenium level and esophageal cancer: An ecological study in a high risk area for esophageal cancer

ProjectGolestan province, located in northeast of Iran, has been known as a high risk area for esophageal cancer (EC). This study was conducted to assess the relationship between soils selenium (Se) level and development of EC in this region.ProceduresIn this ecological study, 135 blocks were identified in Golestan province based on geographical altitude and longitude on the map. One soil sample was collected from the center of each block. Then we investigated Se concentration in soil samples by flame atomic absorption spectrometry. Statistical analysis was performed by the Pearson correlation test and Student t-tests. P-values of less than 0.05 were considered as significant.ResultsThe mean±SD of soils Se level in Golestan province was 3.7±1.61 mg/kg. There was a positive correlation between soils Se level and EC rates in this area (P=0.03) (Pearson correlation coefficient=0.19). Soils Se concentration was significantly higher in high (4.13 mg/kg) than in the low (3.39 mg/kg) EC rate areas (P=0.01).ConclusionsWe found high soils Se concentration and a significant positive relationship between soils Se level and EC rate in Golestan province of Iran. So, high soils Se level may play a possible role in developing EC in this area, specifically in Turkmensahra (very high EC rates).     

Cottonseed Meal Bioactive Peptides as an Alternative to Antibiotic Growth Promoters in Broiler Chicks

The aim of this experiment was to evaluate the effects of bioactive peptides derived from enzymatic hydrolysis of cottonseed meal (CSBP) compared with zinc bacitracin, as an antibiotic growth promoter (AGP), on productive traits, serum lipid profile, and ileal microbial population in broiler chickens. A total number of 240-day-old broiler chicks (Ross 308) were allocated into 4 treatments, replicated 5 times based on a completely randomized design. The dietary treatments included a basal diet serving as control group, basal diet?+?40 mg/kg zinc bacitracin as AGP group, and the basal diet supplemented with 15 or 20 g/kg CSBP substituting equal quantity of maize and soybean meal. Performance traits, including daily weight gain, feed intake, feed conversion ratio (FCR), and livability were recorded. At the end of the study, serum lipid parameters, ileal microbial population, and economical indices were determined. The results indicated that feed intake and FCR increased (P?<?0.05) in birds receiving 20 g/kg CSBP over the entire period (1–35 days), but there was no significant effect of CSBP on body weight, although numerically higher than the control group. However, the antibiotic group showed a significant increase (P?<?0.05) in body weight and feed intake. Livability, European Production Efficiency Factor (EPEF), and European Broiler Index (EBI) significantly improved in broiler chickens fed antibiotic and 15 g/kg CSBP supplement (P?<?0.05). Adding 20 g/kg of CSBP to the diet significantly increased serum triglycerides and decreased low-density lipoproteins (LDL) compared to the control and antibiotic groups on day 35. The relative weight of abdominal fat and LDL to HDL ratio were significantly lower for CSBP and antibiotic treatments than the control group (P?<?0.05). Supplementation of antibiotic and both graded levels of CSBP decreased the ileum population of Escherichia coli (P?<?0.05). The current findings suggest that including CSBP in broiler diets may benefit production through improving growth rate of broilers and balancing gut microbiota population. In addition, CSBP could be considered as a potential alternative to antibiotics in an AGP free production system.

     

Saturated fat stimulates obesity and hepatic steatosis and affects gut microbiota composition by an enhanced overflow of dietary fat to the distal intestine

We studied the effect of dietary fat type, varying in polyunsaturated-to-saturated fatty acid ratios (P/S), on development of metabolic syndrome. C57Bl/6J mice were fed purified high-fat diets (45E% fat) containing palm oil (HF-PO; P/S 0.4), olive oil (HF-OO; P/S 1.1), or safflower oil (HF-SO; P/S 7.8) for 8 wk. A low-fat palm oil diet (LF-PO; 10E% fat) was used as a reference. Additionally, we analyzed diet-induced changes in gut microbiota composition and mucosal gene expression. The HF-PO diet induced a higher body weight gain and liver triglyceride content compared with the HF-OO, HF-SO, or LF-PO diet. In the intestine, the HF-PO diet reduced microbial diversity and increased the Firmicutes-to-Bacteroidetes ratio. Although this fits a typical obesity profile, our data clearly indicate that an overflow of the HF-PO diet to the distal intestine, rather than obesity itself, is the main trigger for these gut microbiota changes. A HF-PO diet-induced elevation of lipid metabolism-related genes in the distal small intestine confirmed the overflow of palm oil to the distal intestine. Some of these lipid metabolism-related genes were previously already associated with the metabolic syndrome. In conclusion, our data indicate that saturated fat (HF-PO) has a more stimulatory effect on weight gain and hepatic lipid accumulation than unsaturated fat (HF-OO and HF-SO). The overflow of fat to the distal intestine on the HF-PO diet induced changes in gut microbiota composition and mucosal gene expression. We speculate that both are directly or indirectly contributive to the saturated fat-induced development of obesity and hepatic steatosis.     

Differences in fennel seed responses to drought stress at the seed formation stage in sensitive and tolerant genotypes

To understand the effects of drought on fennel seed production and determine the underlying molecular processes, various fennel genotypes were exposed to drought stress. The yield and quality, including aromatic oil content, of fennel seeds were reduced by drought during seed development. To explore drought-induced biological processes in fennel, a label-free/gel-free proteomic analysis was performed. In Gaziantep and Tatmaj cultivars, which are sensitive and tolerant fennel genotypes, respectively, 106 and 92 drought-responsive proteins were identified. Comparison of protein-functional profiles indicated that proteins classified in stress, cell, and protein synthesis/degradation categories consisted important responsive mechanisms against drought stress. Pathway analysis visualized that the tricarboxylic acid cycle is important for both cultivars. In Tatmaj, moderate activation of proteins related to oxidative pentose phosphate pathway was detected along with an increase in photosynthesis-related proteins. Furthermore, cluster analysis of drought-responsive proteins using protein abundance at milky, dough, and mature stages identified protein homeostasis as a mechanism of drought tolerance in fennel. These results suggest that coordinated energy consumption and supply might be a drought-tolerance mechanism in fennel plants.

     

Structural, functional and molecular analysis of the effects of aging in the small intestine and colon of C57BL/6 J mice

ABSTRACT: BACKGROUND: By regulating digestion and absorption of nutrients and providing a barrier against the external environment the intestine provides a crucial contribution to the maintenance of health. To what extent aging-related changes in the intestinal system contribute to the functional decline associated with aging is still under debate. METHODS: Young (4 M) and old (21 M) male C57BL/6 J mice were fed a control low-fat (10E%) or a high-fat diet (45E%) for 2 weeks. During the intervention gross energy intake and energy excretion in the feces were measured. After sacrifice the small and large intestine were isolated and the small intestine was divided in three equal parts. Swiss rolls were prepared of each of the isolated segments for histological analysis and the luminal content was isolated to examine alterations in the microflora with 16S rRNA Q-PCR. Furthermore, mucosal scrapings were isolated from each segment to determine differential gene expression by microarray analysis and global DNA methylation by pyrosequencing. RESULTS: Digestible energy intake was similar between the two age groups on both the control and the high-fat diet. Microarray analysis on RNA from intestinal scrapings showed no marked changes in expression of genes involved in metabolic processes. Decreased expression of Cubilin was observed in the intestine of 21-month-old mice, which might contribute to aging-induced vitamin B12 deficiency. Furthermore, microarray data analysis revealed enhanced expression of a large number of genes involved in immune response and inflammation in the colon, but not in the small intestine of the 21-month-old mice. Aging-induced global hypomethylation was observed in the colon and the distal part of the small intestine, but not in the first two sections of the small intestine. CONCLUSION: In 21-month old mice the most pronounced effects of aging were observed in the colon, whereas very few changes were observed in the small intestine.     

Effects of human placenta-derived mesenchymal stem cells with NK4 gene expression on glioblastoma multiforme cell lines

Poor prognosis and low survival are commonly seen in patients with glioblastoma multiforme (GBM). Due to the specific nature of solid tumors such as GBM, delivery of therapeutic agents to the tumor sites is difficult. So, one of the major challenges in the treatment of these tumors is a selection of appropriate method for drug delivery. Mesenchymal stem cells (MSCs) have a unique characteristic in migration toward the tumor tissue. In this regard, the present study examined the antitumor effects of manipulating human placenta-derived mesenchymal stem cells (PDMSCs) with NK4 expression (PDMSC-NK4) on GBM cells. After separation and characterization of PDMSCs, these cells were transduced with NK4 which was known as the antagonist of hepatocyte growth factor (HGF). The results indicated that engineered PDMSCs preferably migrate into GBM cells by transwell coculture system. In addition, the proliferation of the GBM cells significantly reduced after coculture with these cells. In fact, manipulated PDMSCs inhibited growth of tumor cells by induction of apoptosis. Our findings suggested that besides having antitumor effects, PDMSCs can also be applied as an ideal cellular vehicle to target the glioblastoma multiforme.     

Synergistic Effect of Fadrozole and Insulin-Like Growth Factor-I on Female-To-Male Sex Reversal and Body Weight of Broiler Chicks

The aim of this study was to investigate the effects of Fadrozole hydrochloride and recombinant human insulin-like growth factor I (rhIGF-I) on female-to-male sex reversal, hatching traits, and body weight of broiler chickens. On the third day of incubation, fertile eggs were randomly assigned to five experimental groups comprising (i) Fadrozole (0.1 mg/egg), (ii) rhIGF-I (100 ng/egg), (iii) Fadrozole (0.1 mg/egg) + rhIGF-I (100 ng/egg), (iv) vehicle injection (10 mM acetic acid and 0.1% BSA), and (v) non-injected eggs. Eggs in the rhIGF-I-injected groups showed the mode of hatching time at the 480th hour of incubation, 12 hours earlier compared to the other groups, with no statistically significant difference in mortality and hatchability. On Day 1 and 42 of production, 90% of genetically female chicks were masculinized using Fadrozole treatment, while 100% female-to-male phenotypic sex reversal was observed in the Fadrozole+rhIGF-I group. Fadrozole equalized the body weight of both genders, although rhIGF-I was effective on the body weight of male chicks only. Interestingly, combined rhIGF-I and Fadrozole could increase the body weight in both sexes compared to the individual injections (P<0.05). These findings revealed that (i) IGF-I-treated chicken embryos were shown to be an effective option for overcoming the very long chicken deprivation period, (ii) the simultaneous treatment with Fadrozole and IGF-I could maximize the female-to-male sex reversal chance, (iii) the increase in the body weight of masculinized chickens via Fadrozole could be equal to their genetically male counterparts, and (iv) the IGF-I effectiveness, specifically along with the application of aromatase inhibitors in female chicks, indicates that estrogen synthesis could be a stumbling block for the IGF-I action mechanism in female embryos.