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Abstract Background As an in vitro model porcine peripheral blood mononuclear cells (PBMCs) is frequently used as for immunogenetic research with the stimulation of bacterial antigens. To investigate the immunocompetence of PBMCs for recognition of Gram-positive and Gram-negative bacteria and in order to dissect the pathogenesis of diseases, gene expression assay is most commonly used. The gene expressions are required to normalize for reference genes which have tremendous effect on the results of expression study. The reference genes should be stably expressed between different cells under a variety of experimental conditions, but recent influx of data showed that expression stability of reference genes are varied under different experimental conditions. But data regarding the expression stability of reference genes in porcine PBMCs are limited. Therefore, this study was aimed to know whether the expression stability of commonly used reference genes in PBMCs is affected by various bacterial antigens under different experimental conditions in pigs. Results The mRNA expression stability of nine commonly used reference genes (B2M, BLM, GAPDH, HPRT1, PPIA, RPL4, SDHA, TBP and YWHAZ) was determined by RT-qPCR in PBMCs that were stimulated by LPS and LTA in vitro as well as cells un-stimulated control and non-cultured were also consider for this experiment. mRNA expression levels of all genes were found to be affected by the type of stimulation and duration of the stimulation (P Conclusion There was discrepancy in the ranking order of reference genes obtained by different analysing algorithms (geNorm and NormFinder). In conclusion, the geometric mean of the RPL4, B2M and PPIA seemed to be the most appropriate combination of reference genes for accurate normalization of gene expression data in porcine PBMCs without knowing the type of bacterial pathogenic status of the animals and in the case of mixed infection with Gram-negative and Gram-positive bacteria. In case of PBMCs without any stimulation, PPIA, BLM and GAPDH could be suggested as suitable reference genes.

The distribution patterns of desmin and vimentin were determined in Zavot cattle skin using standard immunohistochemical techniques. Skin samples were collected from the different regions of the bodies. Vimentin-immunoreactivity was observed in the cells of subepidermal region, in the myoepithelial cells surrounding sebaceous and sweat glands, in the fibroblast, smooth muscle, endothelial cells of vessels and some hair follicles. Desmin-immunoreactivity was dense in the smooth muscle of arrectores pilorum muscles and bulbus pilorum of the some hair follicles. Desmin and vimentin immunoreactivities were not different among the various regions of Zavot cattle skin. [ABSTRACT FROM AUTHOR]

Examines the effects of In Ovo rh insulin-like growth factors-1 administration on expression of the growth hormone secreragogue receptor (GHSR) during chicken embryonic development. Control of growth hormone secretion; Use of the reverse-transcriptase polymerase chain reaction to determine the expression pattern of GHSR during embryonic development; Isolation of total RNA.

Determines the distinct zonation changes in the adrenal cortex of geese in various ages using both electron and light microscopy. Analysis of the cytoplasm of interrenal cells located beneath the adrenal capsule; Examination of sub-capsular zone (SCZ); Arrangement of SCZ cells.

The purpose of the present study was to determine the distribution of cells containing glucagon (A), insulin (B), and somatostatin (D) in the goose pancreas. Small pieces of tissue from 5 adult geese (Anser anser) pancreases were dissected under deep ether anaesthesia. Sections were stained with Crossmon's connective tissue stain for general observations and Gomori's method for pancreatic islet cells. Sections were further processed for standard immunohistochemical techniques using the avidin-biotin-complex method for the distribution of A, B and D cells. There were no B cells inside the A islets, which were mainly composed of A cells. B islets were mainly composed of insulin-producing B cells arranged in a circular fashion around blood capillaries. Granules containing insulin were localised in blood vessel sites of the B cells. Although D cells were distributed in almost all sites of the A islets, they were mainly localised at the periphery of the B islets. D cells throughout the pancreas were more common than that of B and A cells. The distribution of cells containing insulin, glucagon and somatostatin in the goose pancreas, with small differences, was similar to that of other avian species. [ABSTRACT FROM AUTHOR]

Deals with a study which determined the ontogeny of insulin-like growth factor (IGF)-I, IGF-II, transforming growth factor-β2, and fibroblast growth factor gene expression with regard to embryonic and chicken skeletal muscle development. Materials and methods; Results; Discussion.

The objective of the study was to evaluate the impact of in ovo administration of recombinant human insulin-like growth factor-I (rhIGF-I) on myostatin and transforming growth factor-beta2 (TGF-beta2) gene expression during chicken embryogenesis with emphasis on skeletal muscle development. Eggs were injected once with 100 ng rh IGF-I in 10 mM acetic acid, 0.1% BSA per embryo on day 3 of embryonic development. Total RNA was isolated from whole embryos on each of embryonic days (E) 0 to 6 (n = 6 per day/per treatment), from thoracic/abdominal halves of the embryo at E 7 to 8 (n = 6 per day/per treatment), and from pectoralis muscle tissues at E 9 to 20 (n = 4 per day/per treatment). Reverse-transcription polymerase chain reaction (RT-PCR) was used to synthesize cDNAs. Myostatin mRNA isolated from pectoralis muscles of the rhIGF-I treated group increased on E 10 (approximately 2.5 fold) and remained high through E 13, whereas myostatin mRNA from control pectoralis muscles increased at E 9 and remained high until E 12. TGF-beta2 gene expression from in ovo rhIGF-I treated pectoralis muscles dramatically increased at E 13 (approximately 2.5 fold), in contrast to E 14 from control pectoralis muscle, and gradually declined through E 16. Our results demonstrate that in ovo administration of rhIGF-I on E 3 may alter developmental expression patterns of myostatin and TGF-beta2 genes.