mRNA Expression Of Other Genes Research Articles

Changes in the testicular histomorphometry and their association with genes expression pattern of testes from birth to puberty in Beetal goat kids

In the present study, twenty-eight (n = 28) longitudinally rear male goat kids were slaughtered at monthly intervals (n = 4/group) till the 6th month of age; to examine the testicular development and establishment of spermatogenesis through histo-architecture along with mRNA expression of related genes. The results showed that seminiferous tubular diameter was linearly increased (P < 0.001) at birth (40 µm) to 6th months (200 µm). The epithelial height was 18.41 ± 0.65 µm at birth and 164 ± 2.82 µm at 6th months. Whereas, at 3rd months age greatest increase was observed (74.08 ± 1.66 µm), possibly due to a rapid increase in the numbers of spermatocytes and spermatids (4.0 ± 0.3 and 0.9 ± 0.04, respectively). At the same time point (4–6 months of age), a dramatic increase (P < 0.001) was observed in the mRNA level of genes expressed in haploid germ cells including Protamine (PRM1), Outer dense fiber protein (ODF2) and Stimulated by Retinoic Acid (STRA8). In addition, the maturation of Sertoli cells (SCs) was appeared to be completed at the start of 1st month (5.6 ± 0.5), which was further validated by sudden repression (P < 0.001) of SC-specific SOX9 and FSHR mRNA levels during this period. The spermatids that appeared at the end of the 3rd month reflect the start of spermatogenesis. However, a significant (P < 0.001) number of elongated spermatids and mature spermatozoa was observed at the end of 6 months, which indicated pubertal onset (9.8 ± 0.7 and 9.9 ± 0.9, respectively). The mRNA expression of other genes altered during this developmental phase was Heat-shock Protein 8 (HSPA8) and Aromatase (CYP19A1), both was important in sperm maturation and testosterone level. In conclusion, marked changes in cellular and molecular events were observed from the 3rd month; when active spermatogenesis initiated, and mature spermatozoa appeared in the 6th month at pubertal onset in Beetal bucks.

Use of aspirin to intentionally induce gastrointestinal tract barrier dysfunction in feedlot cattle.

Stress negatively affects the gastrointestinal tract (GIT) barrier function, resulting in compromised animal health. A deeper understanding of how diet and stress impacts the GIT barrier function in feedlot cattle is needed. Aspirin decreases mucus production and mucosal repair in the GIT and could be used as a model for GIT barrier dysfunction research. The objective of this study was to evaluate the effectiveness of aspirin to induce GIT barrier dysfunction in beef cattle. In experiment 1, sixteen crossbred heifers (425.0 ± 8.6 kg) were allotted to 0, 50, 100, or 200 mg/kg body weight (BW) aspirin doses based on BW. Experiment 1 consisted of two periods separated by 4 wk where four heifers per treatment received the same aspirin dose during each period. Heifers were fed a 49.4% corn silage and 50.6% concentrate diet. The 200 mg/kg BW aspirin treatment was dosed as a 100 mg/kg BW aspirin oral bolus 36 and 24 h prior to Cr-ethylenediaminetetraacetic acid (EDTA) dosing (1 liter; 180 mM). The 50 and 100 mg/kg BW aspirin treatments were dosed as an oral bolus 24 h prior to Cr-EDTA dosing. Urine was collected every 3 h for 48 h and analyzed for Cr. Serum was collected at 0 and 48 h and analyzed for lipopolysaccharide-binding protein (LBP), interleukin-6, serum amyloid A (SAA), haptoglobin, and aspartate aminotransferase. In experiment 2, sixteen crossbred steers (576.0 ± 14.2 kg) fed a similar diet were allotted by BW to the 0 and 200 mg/kg BW aspirin treatments (eight steers/treatment) and were slaughtered 24 h after the last dose. Jejunal tissues were collected, and claudin (CLDN) 1, 2, and 3, occludin, and zonula occludens tight junction messenger ribonucleic acid (mRNA) expression was determined. Data were analyzed using the MIXED procedure of SAS. Urinary Cr excretion increased linearly at hours 3, 6, 9, and 12 (P ≤ 0.04) as aspirin dose increased from 0 to 200 mg/kg. Aspirin linearly increased Cr absorption (P = 0.02) and elimination (P = 0.04) rates and linearly decreased mean retention time of Cr (P = 0.02). Aspirin increased SAA (P = 0.04) and tended to increase LBP (P = 0.09) in serum but did not affect any other serum inflammatory marker (P ≥ 0.19). Aspirin tended to increase jejunal CLDN-1 mRNA expression (P = 0.10) but did not affect the mRNA expression of other genes regulating tight junction function (P ≥ 0.20). Results from this study indicate that aspirin disrupts the GIT barrier function in beef cattle and has a potential as a model in GIT permeability research.

The effects of phospholipase C on oestradiol and progesterone secretion in porcine granulosa cells cultured in vitro.

Granulosa cells play important roles in the regulation of ovarian functions. Phospholipase C is crucial in several signalling pathways and could participate in the molecular mechanisms of cell proliferation, differentiation and ageing. The objective of this study was to identify the effects of phospholipase C on the steroidogenesis of oestradiol and progesterone in porcine granulosa cells cultured in vitro. Inhibitor U73122 or activator m-3M3FBS of phospholipase C was added to the in vitro medium of porcine granulosa cells, respectively. The secretion of oestradiol decreased after 2hr, 8hr, 12hr, 24hr and 48hr of treatment with 500nM U73122 (p<.05) and decreased after 2hr of treatment in the 500nM m-3M3FBS addition group (p<.05). The secretion of progesterone increased after 4hr of treatment with 500nM U73122 (p<.05) and increased after 2hr and 8hr of treatment in the 500nM m-3M3FBS addition group (p<.05). The ratio of oestradiol to progesterone decreased at each time point, except 8hr after the addition of 500nM U73122 (p<.05). The ratio of oestradiol to progesterone decreased after 2hr (p<.05) of treatment with 500nM m-3M3FBS. In genes that regulate the synthesis of oestradiol or progesterone, the mRNA expression of CYP11A1 was markedly increased (p<.05), and the mRNA expression of other genes did not change significantly in the U73122 treatment group, while the addition of m-3M3FBS did not change those genes significantly despite the contrary trend. Our results demonstrated that phospholipase C can be a potential target to stimulate the secretion of oestradiol and suppress progesterone secretion in porcine granulosa cells cultured in vitro, which shed light on a novel biological function of phospholipase C in porcine granulosa cells.

Non-canonical NF-κB Plays a Pivotal Role in Non-Hodgkin's Lymphoma.

This study tried to dissect BAFF/BAFF-R-mediated non-canonical NF-κB signaling in the drug resistance of B-cell non-Hodgkin's lymphoma. FQ-PCR was employ to determine the mRNA expression of BAFF, BAFF-R, Bcl-xL, and RIPK2 in the circulating blood of B-cell non-Hodgkin's lymphoma patients. Further correlation studies were performed with the gene expression in the circulating blood and tumor tissue. MTT assay as used to determine BAFF's role on lymphoma cell proliferation. Western blot was employed to determine protein expression after BAFF stimulation. The mRNA expression of BAFF, BAFF-R, Bcl-xL, and RIPK2 in the circulating blood of the resistant group was higher than that of the non-resistant group, which was statistically significant. The mRNA expression of the target genes was positively correlated. The mRNA expression was positively correlated with disease progression, which was statistically significant. The Bcl-xL mRNA expression in the resistant group was relatively higher than that in the non-resistant group, which was also statistically significant. However, the mRNA expression of other genes only showed increased tendency compared with non-resistant group. There was no significant change between target genes in different tumor tissues. With increased BAFF concentration and prolonged exposure, the proliferation of the tumor cells increased significantly, which was statistically significant. Western blot showed the expression of BAFF, BAFF-R, Bcl-xL, and RIPK2 all increased with increased BAFF concentration, which was also statistically significant. In B-cell, non-Hodgkin's lymphoma, BAFF may activate non-canonical NF-κB signaling to regulate drug resistance.

18F]Fluorodeoxyglucose accumulation as a biological marker of hypoxic status but not glucose transport ability in gastric cancer

BackgroundThe use of [18F] 2-fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) for detection of gastric cancer is often debated because FDG uptake varies for each patient. The purpose of this study was to clarify the molecular mechanisms involved in FDG uptake.Material and methodsFifty patients with gastric cancer who underwent FDG-PET and gastrectomy were studied. Snap-frozen tumor specimens were collected and examined by real-time PCR for relationships between maximum standardized uptake value (SUV) and mRNA expression of the following genes: glucose transporter 1 (GLUT1), hexokinase 2 (HK2), hypoxia-inducible factor 1α (HIF1α), and proliferating cell nuclear antigen (PCNA).ResultsTumor size was the only clinicopathological parameter that significantly correlated with SUV. Transcripts for the genes evaluated were about three-fold higher in malignant specimens than in normal mucosa, although only HIF1α was significantly correlated with SUV. When divided into intestinal and non-intestinal tumors, there was a significant correlation between SUV and tumor size in intestinal tumors. Interestingly, the weak association between SUV and HIF1α expression in intestinal tumors was substantially stronger in non-intestinal tumors. No correlation was found between SUV and mRNA expression of other genes in intestinal or non-intestinal tumors.ConclusionSUV was correlated with HIF1α, but not PCNA, HK2, or GLUT1 expression. FDG accumulation could therefore represent tissue hypoxia rather than glucose transport activity for aggressive cancer growth.

Nucleoside analog ARC targets Mcl-1 to induce apoptosis in leukemia cells

Nucleoside analog ARC targets Mcl-1 to induce apoptosis in leukemia cells

Gene networks regulating lipid synthesis in bovine mammary gland during the lactation cycle

Regulation of lipid synthesis in the bovine mammary gland is a complex dynamic process likely controlled by interacting gene networks. Little is known about these sensing and signal transduction circuits. Mammary biopsies from 6 Holstein cows were obtained at -15, 1, 15, 30, 60, 120, and 240 days relative to parturition (DIM), and RNA used for qPCR of ABCA1, ABCG2, ACACA, ADFP, BTN1A1, CD36, DGAT1, ESR1, FABP3, FADS1, FASN, GPAM, INSIG1, INSIG2, LPIN1, LPL, PPARG, PPARGC1A, PPARGC1B, RETN, SCAP, SCD, SREBF1, SREBF2, and THRSP. These genes are relatively well-studied in non-ruminant cells and tissues due to their key roles in aspects of carbohydrate and lipid metabolism. Except for PPARGC1B, lactation increased (P < 0.01) mRNA expression of other genes by 1.5 (ABCA1) to 80-fold (FABP3, LPL) relative to day -15. Peak expression patterns were observed at 60 DIM for all genes except PPARG, PPARGC1A, SCAP, and THRSP, which peaked (3-, 10-, 1.7-, and 3-fold, respectively) at 120 DIM. Network analysis using Ingenuity Pathway Analysis revealed regulatory loops consisting of SREBF2, SREBF1, PPARG, PPARGC1A, and PPARGC1B along with their down-stream targets (e.g. SCD, ACACA, INSIG1, FABP3). LPIN1, INSIG2, and SCAP appeared to independently regulate expression of several lipogenic genes. LPIN1 (25-fold higher on day 60) and AGPAT6 (16-fold higher on day 60) seem to play a role in bovine milk fat synthesis regulation. Results suggest that mammary lipid synthesis is regulated by complex gene networks including transcription factors and novel genes.