Lipins play dual function in lipid metabolism by serving as phosphatidate phosphatase and transcriptional co-regulators of gene expression. Mammalian lipin proteins consist of lipin1, lipin2, and lipin3 and are encoded by their respective genes Lpin1, Lpin2, and Lpin3. To date, most studies are concerned with Lpin1, only a few have addressed Lpin2 and Lpin3. Ontogenetic expression of Lpin2 and Lpin3 and their associations with traits would help to explore their molecular and physiological functions in sheep. In this study, 48 animals with an equal number of males and females each for both breeds of fat-tailed sheep such as Guangling Large Tailed (GLT) and Small Tailed Han (STH) were chosen to evaluate the ontogenetic expression of Lpin2 and Lpin3 from eight different tissues and months of age by quantitative real-time polymerase chain reaction (PCR). Associations between gene expression and slaughter and tail traits were also analyzed. The results showed that Lpin2 mRNA was highly expressed in perirenal and tail fats, and was also substantially expressed in liver, kidney, reproductive organs (testis and ovary), with the lowest levels in small intestine and femoral biceps. Lpin3 mRNA was prominently expressed in liver and small intestine, and was also expressed at high levels in kidney, perirenal and tail fats as well as reproductive organs (testis and ovary), with the lowest level in femoral biceps. Global expression of Lpin2 and Lpin3 in GLT both were significantly higher than those in STH. Spatiotemporal expression showed that the highest levels of Lpin2 expression occurred at 10 months of age in two breeds of sheep, with the lowest expression at 2 months of age in STH and at 8 months of age in GLT. The greatest levels of Lpin3 expression occurred at 4 months of age in STH and at 10 months of age in GLT, with the lowest expression at 12 months of age in STH and at 8 months of age in GLT. Breed and age significantly influenced the tissue expression patterns of Lpin2 and Lpin3, respectively, and sex significantly influenced the spatiotemporal expression patterns of Lpin3. Meanwhile, Lpin2 and Lpin3 mRNA expression both showed significant correlations with slaughter and tail traits, and the associations appear to be related with the ontogenetic expression as well as the potential functions of lipin2 and lipin3 in sheep.
This study is to evaluate the effect of different levels of ensiling density on the fermentation quality of guineagrass silages during the early stage of ensiling. Guineagrass at the milky ripe stage was chopped and ensiled into a small-scale laboratory silo at two ensiling density levels (high density at 95 g/silo and low density at 75 g/silo). Three silos per level were opened after six ensiling periods (0.5, 1, 1.5, 2, 3 and 7 days of ensiling) and the fermentation qualities were analyzed. Within the initial 1.5 days of ensiling there were not significant (p>0.05) differences in the fermentation qualities between two density levels silages, and an almost constant pH and no or only small amounts of lactic acid, acetic acid and total volatile fatty acids were detected. However, the high density silage significantly (p<0.05) increased the rate and extent of fermentation after 1.5 days of ensiling, which was well reflected in significantly (p<0.05) faster and larger pH decline and lactic acid production at each elapsed time as compared with the low density silage. This resulted in significantly (p<0.05) lower finial pH and significantly (p<0.05) higher lactic acid content at the end of the experiment. Moreover, there was higher AA content relative to LA in both the H-D and L-D silages during the full fermentation course, and resulted in the AA-type silage. There were generally somewhat or significantly (p<0.05) higher acetic acid, volatile fatty acids and ammonia-N/total nitrogen in the high density silage than in the low density silage during the initial 3 days of ensiling. However, there were higher (p>0.05) ammonia-N/total nitrogen and significantly (p<0.05) higher butyric acid content in the low density silage at day 7 of ensiling. The silages of two density levels showed an initial increase in glucose between 0.5 and 1 day for the high density silage and between 1 and 1.5 days for the low density silage, respectively, thereafter showed a large decrease until the end of the experiment. There were not large differences (p>0.05) in ethanol content between the low density and high density silages that showed small amounts within initial 3 days of ensiling. However, the low density silage had a significantly (p<0.05) higher ethanol content than the high density silage at the end of experiment. From the above results it was suggested that the increase in ensiling density was an effective method to improve the fermentation quality, especially for tropical grasses.
Objective: This study aimed to examine the effect of sodium butyrate (SB) on growth performance, immune status, organs weights, and microarchitecture of lymphoid organs and small intestine. Methods: A total of 120, 1-d-old broiler chicks were distributed into the following four treatment groups: corn-soy based basal diet (BD) without supplement (control), or the same BD supplemented with 0.1 g/kg zinc bacitracin (ZnB), 0.5 g/kg SB (SB-0.5), or 1.0 g/kg SB (SB-1), respectively. Six birds/group were killed on d-21 and d-35, and samples were collected. Results: Cell-mediated immune response at 48 h post-Phytohemagglutinin-P injection, and antibody titer against Newcastle disease vaccine and sheep red blood cells on d-35 was noted higher (p<0.05) in SB-1 compared to ZnB and control. Lower (p<0.05) feed conversion ratio (FCR) was attained by the supplemented groups. Thymus and spleen weighed more (p<0.05) in SB-1, and bursa registered more (p<0.05) weight in both SB groups compared to control. On d-21, areas of thymus medulla and spleen germinal centers were noted higher (p<0.05) in SB-1 group. The villus height and villus surface area increased (p<0.05) in duodenum and jejunum in both SB groups on d-21, and in SB-1 on d-35, respectively compared to ZnB and control. On d-21, number of goblet cells containing mucins of acidic nature increased (p<0.05) in all the segments of small intestines in SB-1 group compared to control, and on d-35 in ileum compared to other groups. Conclusion: In conclusion, SB improved growth performance and immunity as well as modulated morphology of lymphoid organs and gut mucosa in broiler chickens.
Objective: Selenium-independent glutathione peroxidase (GPx5) is specifically expressed in the mammalian epididymis and plays an important role in protecting sperm from reactive oxygen species and lipid peroxidation damage. This study investigates GPx5 expression in the epididymis of Small Tail Han sheep. Methods: GPx5 expression was studied in three age groups: lamb (2 to 3 months), young (8 to 10 months), and adult (18 to 24 months). The epididymis of each age group divided into caput, corpus and cauda, respectively. Analysis the expression quantity of GPx5 in epididymis and testis by real-time fluorescent quantitative polymerase chain reaction and Western blot. Finally, GPx5 protein locating in the epididymis by immunohistochemical. Results: The results demonstrate that in the lamb group, the GPx5 mRNA, but not protein, can be detected. GPx5 mRNA and expressed protein were detected in both the young and adult groups. Moreover, both the mRNA and protein levels of GPx5 were significantly higher in the young group than in other two groups. When the different segments of epididymis were investigated, GPx5 mRNA was expressed in each segment of epididymis regardless of age. Additionally, the mRNA level in the caput was significantly higher than that in corpus and cauda within same age group. The GPx5 protein was in the epithelial cells' cytoplasm. However, GPx5 mRNA and protein were not detected in the testis. Conclusion: These results suggest that GPx5 is mainly expressed in the epididymis of Small Tail Han sheep, and that the expression level of GPx5 is associated with age. Additionally, GPx5 was primarily expressed in the epithelial cells of the caput. Taken together, these studies indicate that GPx5 is expressed in the epididymis in all age grades.
Objective: The present study was to investigate the association of polymorphisms in exon-9 of the bone morphogenetic protein receptor-1B (BMPR-1B) gene (C864T) with litter size in 240 Dorset, 232 Mongolian, and 124 Small Tail Han ewes. Methods: Blood samples were collected from 596 ewes and genomic DNA was extracted using the phenol: chloroform extraction method. The 304-bp amplified polymerase chain reaction product was analyzed for polymorphism by single-strand conformation polymorphism method. The genotypic frequency and allele frequency of BMPR-1B gene exon-9 were computed after sequence alignment. The ${\chi}^2$ independence test was used to analyze the association of genotypic frequency and litter size traits with in each ewe breed, where the phenotype was directly treated as category. Results: The results indicated two different banding patterns AA and AB for this fragment, with the most frequent genotype and allele of AA and A. Calculated Chi-square test for BMPR-1B gene exon-9 was found to be more than that of p value at the 5% level of significance, indicating that the population under study was in Hardy-Weinberg equilibrium for all ewes. The ${\chi}^2$ independence test analyses indicated litter size differences between genotypes was not the same for each breed. The 304-bp nucleotide sequence was subjected to BLAST analysis, and the C864T mutation significantly affected litter size in singletons, twins and multiples. The heterozygosity in exon-9 of BMPR-1B gene could increase litter size for all the studied ewes. Conclusion: Consequently, it appears that the polymorphism BMPR-1B gene exon-9 detected in this study may have potential use in marker assisted selection for litter size in Dorset, Mongolian, and Small Tail Han ewes.
The cell cycle phase in which donor nuclei exist prior to nuclear transfer is an important factor governing developmental rates of reconstituted embryos. It was suggested that quiescent G0 and cycling G1 cells could support normal development of reconstituted embryos. In a quest of optimized donor nuclei treatment prior to nuclear transfer, this study was undertaken to examine the cell cycle characteristics of bovine fetal and adult somatic cells when cultured under a variety of culture treatments and the cell cycle change with the lapse of time after trypsinization. This was archived by measuring the DNA content of cells using flow cytometry, Cultured fetal fibroblast cells, adult skin and muscle cells, and cumulus cells were divided by 3 culture treatments; 1) grown to 60-70% confluency (cycling), 2) serum starved culture, 3) culture to confluency. Trypsinized cells were fixed by 70% ethanol and stained with propidium iodide. For one experiment, trypsinized cells were resuspended in DMEM+10% FBS and incubated for 1.5, 3 and 6 h with occasional shaking before ethanol fixation. Cell cycle phases were determined by flow cytometry enabling calculation of percentages of G0+G1, S and G2+M. The majority of cells were in G0+Gl stage regardless of origin of cells. Cultures that were serum starved or cultured to confluency contained significantly (P<0.05) higher percentages of cells in G0+G1 (89.5-95.4%). For every cell lines and culture treatments, percentages of cells in existing in G0+G1 increased with decreasing of the cell size from large to small. In the serum starved and confluency groups, about 98% of small cells were in G0+G1 Serum starved culture contained higher percentages of small-sized cells (38.5-66.9%) than cycling and confluent cultures regardless of cell lines (P<0.05). After trypsinization of fetal fibroblast and adult skin cells that were serum starved and cultured to confluency, the percentages of cells in G0+G1 significantly increased by incubation for 1.5(95.7-99.5%) and 3.0 h (95.9-98.6%). The results suggest that the efficient synchronization of bovine somatic cells in G0+G1 for nuclear transfer can be established by incubation for a limited time period after trypsinization of serum starved or confluent cells.
The Journal of Korean Institute of Electromagnetic Engineering and Science
/
v.11
no.7
/
pp.1194-1203
/
2000
A magnetic field exposure system that generates 60 Hz magnetic fields from 1 mG to 5 G was designed and constructed for small-sized animal study. In order to investigate as many animals as possible at one series of test, uniform magnetic fields are required at wide living area of the animals. In this article, a cubic shaped field exposure system with three animal living floors was designed, which offers about 50 seating capacity. For calculation of magnetic fields inside the cage, a three-dimensional calculation program was developed. Using this, optimum electric current ratio of inner coil to outer coil and position of each coil were determined. Meanwhile, inductance of the exposure system was calculated for the design of power supply. The field measurement results of the manufactured exposure system showed that the difference between maximum and minimum magnetic field at the testing floors was less than 3%, which strongly demonstrated the field exposure system was good for small sized animal study.
The objective of this study was to investigate polymorphisms of BMPRIB (bone morphogenetic protein type IB receptor) gene and its effect on litter size traits in sheep. Three populations including 101 Small Tailed Han sheep, 79 Poll Dorset and 81 hybrids (Poll Dorset${\times}$Small Tailed Han sheep) were used to detect the polymorphisms in exon 6 region of sheep BMPRIB gene. A fragment of approximately 190bp was amplified by one pair of primers, the polymorphism was revealed from the analysis of three populations by the technique of PCR -SSCP, and a mutation from A to G at 746 of the coding region was confirmed by sequencing in several individual. Statistical results indicated the distribution of allele B (with a A$\longrightarrow$G mutation) and A (without mutation) or genotype AA, AB and BB frequencies differed in three populations. BB genotype (44.55%) and B allele (66.34%) frequencies of Small Tailed Han sheep were higher than those of the others. Analysis of variance showed that the polymorphism of BMPRIB gene was associated with positive effect on litter size traits. The means of genotype BB and AB were about 1.04 and 0.74 more than genotype AA for litter size (p<0.05). Analysis of BMPRIB genotype effects on litter size in three populations indicates the existence of genotype BB or B allele increases the litter size. It suggested that the polymorphism in exon 6 (at 746 in the coding region) of sheep BMPRIB gene may be used as a marker for early selection of prolificacy in sheep.
The study was designated to investigate the electron microscopic findings following diethylnitrosamine (DEN) treatment in rats. Forty four male (Srague Dawley) rats were continuously given water containing 0.01% DEN for 13 weeks and livers of five rats with more tumor lesions at 16 and 17 weeks after initial treatment were used as EM materials. In transmission electron microscopic findings, most small-sized hepatocytes were active cells containing large mount of organelles, but light (pale staining) hepatocytes among small-sized hepatocytes were injured cells containg disorganized organelles. Tumor cells among small-sized hepatocytes were irregularly arranged and have pleomorphic nuclei containing electron dense chromatin but the organelles in cytoplasm were swelled. Large-sized hepatocytes were active cells with condensed chromatin but the cytoplasm of these cells were pale due to be injured and dilated organelles. Dark hepatocytes were apoptotic cells with homogenous pyknotic nuclei and cytoplasm, and the cytoplasm of these cells contained dilated smooth endoplasmic reticulum (sER) but these sER were non-vesiculated. Cholangiocarninoma cells were crowded and were pale by far less number of organelles in cytoplasm and nuclei. In scanning electron microscopic findings, the lumens of portal veins, bile canaliculi, bile ductules, bile ducts and sinusoids were dilated and have irregular folded inner surface by protruded parenchyma.
An 8-year-old spayed female Maltese (2.5 kg of body weight) presented with the primary complaint of loud heart murmur and exercise intolerance. Diagnostic imaging revealed severe pulmonic stenosis (peak velocity 5.2 m/s) with right ventricular hypertrophy. The dog revisited after 2 years, at which time, diagnostic imaging revealed severe biventricular hypertrophy, dynamic left ventricular outflow tract obstruction, left atrial dilation and pulmonary hypertension with worsened pre-existing pulmonic stenosis. Postmortem investigation revealed hypertrophic cardiomyopathy and regional myocardial infarction. The case was diagnosed as hypertrophic cardiomyopathy secondary to severe right and left ventricular outflow tract obstruction.
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