NVP-BEZ235 induces autophagy of polycystic kidney rat cholangiocytes

AIM: To explore the effect of dual PI3K/Akt/mTOR inhibitor NVP-BEZ235 on autophagy of polycystic kidney (PCK) rat cholangiocytes. METHODS: The protein levels of p-mTOR and p-Akt in the bile duct epithelial cells were examined by immunohistochemistry. The effect of NVP-BEZ235 on the viability of cholangiocytes was detected by WST-1 assay. The levels of PI3K/Akt/mTOR signaling pathway and autophagy-related proteins with NVP-BEZ235 treatment were determined by Western blot. The effects of LC3 and Beclin 1 silencing, and authophagy-specific inhibitor 3-methyladenine (3-MA) on the cell viability were analyzed by WST-1 assay. RESULTS: The protein levels of p-Akt and p-mTOR were highly increased in the bile duct epithelium of the PCK rats. NVP-BEZ235 significantly inhibited the viability of the cholangiocytes in a dose- and time-dependent manner (P<0.05). NVP-BEZ235 significantly reduced the levels of PI3K/Akt/mTOR signaling pathway-related proteins in the PCK rat cholangiocytes. NVP-BEZ235 upregulated the autophagy-specific proteins LC3 II and Beclin 1. The inhibitory effect of NVP-BEZ235 on the cell viability was weakened by treatment with 3-MA and knockdown of LC3 and Beclin 1 (P<0.01).CONCLUSION: The PI3K/Akt/mTOR inhibitor NVP-BEZ235 suppresses the viability of PCK rat cholangiocytes, and the mechanism is closely related with autophagy.     

Effects of substituting kraft pulp with corn silage on dry matter intake,ruminal mat formation,rumen fermentation,and rumination activity in non‐lactating cows

The present study examined the effects of substituting kraft pulp (KP) with corn silage (CS) on dry matter intake (DMI), ruminal mat and rumen fermentation characteristics, and rumination. Four non‐lactating, rumen‐cannulated Holstein cows were fed a CS diet comprising 36% grass silage (GS) and 64% CS or a KP diet comprising 36% GS, 57% KP, and 7% soybean meal. DMI was significantly lower in cows fed the KP diet than in those fed the CS diet (p < 0.05), whereas rumination time did not significantly differ between the treatments. Dry matter content in the rumen immediately before and 3 h after feeding was significantly higher in cows fed the KP diet than in those fed the CS diet (p < 0.05). The consistency and thickness of the ruminal mat did not significantly differ between the treatments. The ruminal mean retention time of feed particles tended to be longer in cows fed the KP diet than in those fed the CS diet (p < 0.10). The ruminal digestion rate of KP was comparable to that of GS and CS. Because ruminal mat was formed and rumination was stimulated, KP was considered to have the equivalent physical effectiveness as CS.     

Pathogenicity evaluation of Ralstonia pseudosolanacearum race 4 on ginger by leaf-clipping inoculation

Journal of General Plant Pathology - An easy, rapid method has been needed to test the pathogenicity of strains of Ralstonia pseudosolanacearum, the cause of serious bacterial wilt on ginger...     

CO2 efflux from leaf litter focused on spatial and temporal heterogeneity of moisture

Leaf litter respiration (R _LL) was directly measured in situ to evaluate relationships with the water content in leaf litter (WC), which is distributed heterogeneously under natural conditions. To do so, we developed a small, closed static chamber system using an infrared gas analyzer, which can measure instantaneous R _LL. This study focuses on the measurement of CO₂ effluxes from leaf litter using the chamber system in the field and examines the relationship between R _LL and WC among seven broadleaf species in a temperate forest. The measurements focused on the position of leaves within the litter layer, finding that both R _LL and WC were significantly higher in the lower layer. The value of R _LL increased with increasing WC, and the response of R _LL to WC was similar among all seven species. Moreover, the temporal variation in WC differed among three species and was associated with leaf litter thickness. The observed heterogeneity in WC induced by the physical environment (e.g., position and thickness of leaf litter) affects the variation in WC and, therefore, both R _LL and the decomposition rates of organic matter in the litter layer.     

Spatiotemporal distribution of Flavobacterium psychrophilum and ayu Plecoglossus altivelis in rivers revealed by environmental DNA analysis

Outbreaks of bacterial cold-water disease (BCWD), caused by Flavobacterium psychrophilum, are widespread in Japan, especially among ayu Plecoglossus altivelis. There are few investigations of F. psychrophilum in river water, and its seasonal distribution has not been clarified. We aimed to identify the spatiotemporal dynamics of F. psychrophilum and ayu to provide information that is useful for establishing a countermeasure for BCWD. Quantitative analysis of environmental DNA (eDNA) was used to clarify the year-round dynamics of ayu and F. psychrophilum. We sampled river water from the Nagara and Ibi rivers in Japan, and conducted monthly water sampling and eDNA quantification. Changes in the eDNA concentration of ayu were consistent with the known life histories of the fish. There was a strong negative correlation between the eDNA concentration of F. psychrophilum and water temperature, suggesting a strong dependence of F. psychrophilum dynamics in the river on water temperature. Furthermore, relatively high eDNA concentrations were recorded for both organisms in early summer and fall, suggesting that ayu is infected with F. psychrophilum during these seasons when experiencing up- and downmigration, respectively.

     

Intraspecific phenotypic variation associated with nuclear DNA content in Lolium perenne L.

The variation in nuclear DNA content and its association with phenotypic traits were examined in 15 cultivated populations of perennial ryegrass (Lolium perenne L.), including herbage and turf populations. Nuclear DNA contents of 15 populations were measured by a flow cytometric method using DAPI (4′-6-diamidino-2-phenylindole) as a fluorochrome. DNA contents were also measured using PI (propidium iodide) for six populations that showed large differences in DAPI values to confirm the difference. There were significant differences in nuclear DNA contents among the populations for both dyes. Of the total variation in nuclear DNA content measured with DAPI, 29% was ascribed to the inter-population variation, 46% was ascribed to the intra-population variation, 24% was ascribed to the random error variation. Herbage populations tended to have larger DNA content than did turf populations. 2C DNA content was positively correlated with cell size, seed size and single leaf size, although significant correlations were mainly due to a small number of populations with large and small DNA contents. These results suggest that intraspecific variation in nuclear DNA content plays important roles in determining phenotypic differences between cultivated populations of L. perenne. This revised version was published online in August 2006 with corrections to the Cover Date.     

Buckwheat extract inhibits progression of renal failure

Rats subjected to partial resection of the parenchyma showed reduced radical-scavenging activity in the remaining kidney and increased severity of renal tissue lesions. However, in similarly nephrectomized rats given buckwheat extract, the state of oxidative stress improved by restoring the decreased activities of reactive oxygen species-scavenging enzymes such as superoxide dismutase and catalase. The degree of mesangial proliferation, severity of extratubular lesions such as crescents and adhesions, glomerulosclerosis index, and severity of tubular interstitial lesions also improved. In addition, nephrectomized rats given buckwheat extract showed improvement in renal function, as indicated by decreased serum level of creatinine, with a significant decrease in the level of methylguanidine, a uremic toxin produced from creatinine in the presence of hydroxyl radical.     

Antioxidative activities of fractions obtained from brewed coffee

The antioxidative activity of column chromatographic fractions obtained from brewed coffee was investigated to find antioxidants and to assess the benefit of coffee drinking. The dichloromethane extract inhibited hexanal oxidation by 100 and 50% for 15 days and 30 days, respectively, at the level of 5 microg/mL. A GC/MS analysis of fractions, which exhibited oxidative activity, revealed the presence of antioxidative heterocyclic compounds including furans, pyrroles, and maltol. The residual aqueous solution exhibited slight antioxidative activity. The inhibitory activity (%) of the seven fractions from an aqueous solution toward malonaldehde formation from lipid oxidation ranged from 10 to 90 at a level of 300 microg/mL. The results indicate that brewed coffee contains many antioxidants and consumption of antioxidant-rich brewed coffee may inhibit diseases caused by oxidative damages.