MicroRNA-493-5p suppresses colorectal cancer progression via the PI3K-Akt-FoxO3a signaling pathway.

OBJECTIVE: MicroRNA493-5p (miR-493-5p) appears to have an essential role in the abnormal cell proliferation and migration observed in the development and progression of various cancers. However, the function and mechanism of action of miR-493-5p in colorectal cancer (CRC) is unclear. PATIENTS AND METHODS: MiR-493-5p expression was analyzed in CRC patient tissue samples and cell lines by fluorescence quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). SW480 and Caco-2 cells were transfected with miR-493-5p mimics or treated with the phosphoinositide 3-kinase (PI3K) agonist 740Y-P. Cell proliferation was determined by colony formation and cell proliferation assays and cell migration and invasion by transwell migration and wound-healing assays. The Luciferase reporter assay was used to verify the association between the expression of miR-493-5p and PI3K activity. Expression levels of PI3K, protein kinase B(Akt), and forkhead box O 3a (FoxO3a) proteins were measured by Western blot analysis and immunofluorescence assay. RESULTS: MiR-493-5p expression was significantly downregulated in CRC tissue samples and cell lines which was associated with progression of CRC. The proliferation, migration, and invasion of CRC cells were inhibited by miR-493-5p overexpression. The finding that miR-493-5p upregulation decreased PI3K, Akt, and FoxO3a protein expression revealed that it directly targets PI3K. Additionally, the miR-493-5p-mediated suppression of CRC cell proliferation, migration and invasion was counteracted by the PI3K agonist, indicating that miR-493-5p suppresses CRC progression by inhibiting the PI3K-Akt-FoxO3a signaling pathway. CONCLUSIONS: MiR-493-5p suppresses the proliferation, migration, invasion, and progression of CRC via the PI3K-Akt-FoxO3a signaling pathway.

Effects of temperature and temperature shock on the performance and microbial community structure of a submerged anaerobic membrane bioreactor.

Effects of temperature and temperature shock on the performance and microbial community structure of a submerged anaerobic membrane bioreactor (SAnMBR) treating thermomechanical pulping pressate were studied for 416 days. The results showed that the SAnMBR system were highly resilient to temperature variations in terms of chemical oxygen demand (COD) removal. The residual COD in treated effluent was slightly higher at 55 °C than that at 37 and 45 °C. There were no significant changes in biogas production rate and biogas composition. However, temperature shocks resulted in an increase in biogas production temporarily. The SAnMBR could tolerate the 5 and 10 °C temperature shocks at 37 °C and the temperature variations from 37 to 45 °C. The temperature shock of 5 and 10 °C at 45 °C led to slight and significant disturbance of the performance, respectively. Temperature affected the richness and diversity of microbial populations.

Correlation between B lymphocyte abnormality and disease activity in patients with idiopathic membranous nephropathy.

This study investigated the relationship between peripheral blood B lymphocytes, regulatory T-cells and T lymphocyte subsets, the distribution of B lymphocytes in the kidney, and the pathogenesis of idiopathic membranous nephropathy (IMN). Lymphocyte subsets were measured using flow cytometry in 66 patients with clinically-confirmed IMN and in 40 healthy control subjects. Compared with healthy subjects, the number of peripheral blood B lymphocytes was significantly increased in IMN patients and that of regulatory T-cells was significantly decreased, accompanied by an increased CD4(+)/CD8(+) T-cell ratio. There was no relationship between the number of peripheral blood B lymphocytes and markers of kidney function. Although the number of infiltrating B lymphocytes in the kidney of IMN patients was higher, there was no relationship with the number of peripheral blood B lymphocytes. In conclusion, there was no relationship between peripheral blood B lymphocytes and disease activity, suggesting that peripheral blood B lymphocytes are not a biomarker of disease activity and therapeutic efficacy in IMN.

Cloning the swamp buffalo SRY gene for embryo sexing with multiplex-nested PCR.

The polymerase chain reaction (PCR) is an efficient method for sexing embryos. The objective of this study was to develop an accurate and reliable method for sexing swamp buffalo (Bubalus bubalis) embryos. The SRY gene from swamp buffalo genomic DNA was amplified by PCR, using primers based on the sequence of the Holstein SRY gene. This fragment was sequenced based on a BLAST search; the SRY gene was highly conserved. Using a Southern blot, there was a strong signal in genomic DNA only from male swamp buffalo. Two pairs of nested primers, targeted to amplify the swamp buffalo SRY conserved region, were designed for sex identification. Simultaneously, the G3PDH gene was co-amplified to serve as an internal control. A multiplex-nested PCR system was optimized by varying the following individually: concentrations of Mg(2+) and dNTPs, ratio of concentrations of primers and numbers of cycles. Biopsies of 27 IVF-derived embryos and 24 embryos fertilized with Y-chromosome-bearing sperm were examined. Using optimized procedures, clear signals following PCR amplification were obtained from all embryo samples; PCR amplification accuracy was further verified by comparing PCR and dot blots. We concluded that this PCR technique was highly reliable for sexing swamp buffalo embryos.

Glycyl-glutamine-enriched long-term total parenteral nutrition attenuates bacterial translocation following small bowel transplantation in the pig.

BACKGROUND: Improvements in immunosuppression, operative procedure, and posttransplant management have made clinical small bowel transplantation (SBT) feasible. Ischemia and reperfusion injury, total parenteral nutrition (TPN), and devoidment of enteral feeding lead to graft atrophy, gut barrier dysfunction, and bacterial translocation. Glutamine (Gln) is the principal fuel for the enterocyte. The influence of Gln dipeptide-supplemented TPN, especially long-term TPN, on intestinal graft permeability and bacterial translocation is not clear following SBT in the large animal model. Therefore, we studied the effect of glutamine dipeptide, glycyl-glutamine (Gly-Gln), on bacterial translocation following SBT in the pig, which has a physiology similar to humans. MATERIALS AND METHODS: The outbred pigs underwent segmental small bowel autotransplantation and were divided into two groups. In the STPN group (n = 5), the animal received standard TPN devoid of Gly-Gln for 28 days. In the GTPN group (n = 5), the animal received isonitrogenous (0.3 g/kg.day) and isocaloric (33 kcal/kg.day) TPN solution with 2% Gly-Gln for 28 days. RESULTS: At the end of the experiment, Gly-Gln-enriched TPN could maintain the plasma Gln level, graft mucosal Gln and protein concentrations, and skeletal muscle Gln and protein concentrations. Gly-Gln-enriched TPN significantly decreased the bacterial number of mesenteric lymph nodes in the liver and spleen and intestinal permeability to 99mTc-DTPA. There were no significant differences in body weight gain. CONCLUSIONS: The Gly-Gln-enriched long-term TPN may maintain the plasma Gln level, mucosal and muscle Gln, and protein concentrations and attenuate the intestinal permeability to 99mTc-DTPA and bacterial translocation following small bowel transplantation in the pig.

Structure of the nucleocapsid protein gene of rice yellow stunt rhabdovirus.

The complete nucleotide sequence of the N protein gene of rice yellow stunt rhabdovirus (RYSV) was determined by sequencing of cDNA clones derived from the viral genomic RNA. The 3' end of the N gene (messenger sense) was defined by sequence analysis of cDNA clones generated from the N protein mRNA by 3'RACE. The 5' end sequence of the gene was putatively assigned as 5'-AACAC-3'; this sequence is found in the presumed 3' leader/N gene junction region. The mRNA encoding the RYSV N protein is 1714 nt comprising a 15-nt untranslated 5' leader sequence followed by an open reading frame (ORF) of 1563 nt and a 136-nt untranslated 3' region. The calculated molecular mass of the N protein encoded by the ORF is 58,400 Da, which is larger in size than N proteins of other rhabdoviruses. Amino acid composition analysis shows that the RYSV N protein is rather basic with a predicted isoelectric point of 10.04; indeed, a large highly basic region could be found at the carboxy terminal portion of the protein. Amino acid sequence comparison between N proteins of RYSV and sonchus yellow net virus, both of which belong to the same genus Nucleorhabdovirus, revealed an overall 30% identity, with three relatively conserved blocks of 14-20 amino acid residues. Moreover, the hydropathy profiles of the two proteins are generally similar. The structural similarities between the N protein of RYSV and that of lettuce necrotic yellows virus, the type member of the genus Cytorhabdovirus, and those of animal rhabdoviruses, are less significant. Nucleotide sequence determination of 5' and 3' regions flanking the RYSV N gene identified a 14-nt common sequence that is very similar to the consensus gene junction sequences of other plant and animal rhabdoviruses.