Hypoglycemic Effects of a Polysaccharide from Tetrastigma hemsleyanum Diels & Gilg in Alloxan‐Induced Diabetic Mice

Tetrastigma hemsleyanum Diels & Gilg , a well‐known traditional Chinese medicine, possesses antitumor and anti‐inflammatory activity, etc. However, the anti‐diabetic effect has not been determined. In our present study, a water‐soluble polysaccharide, named THP with molecular weight of 93 307 Da, was isolated from T. hemsleyanum by DEAE‐52 ion‐exchange and Sephadex G‐100 chromatography. It contains rhamnose, arabinose, mannose, glucose, and galactose in the molar ratio of 0.07:0.14:0.38:0.21:0.31. Then anti‐diabetic effects of THP were examined by treating alloxan‐induced diabetic mice with different doses (100, 200, and 300 mg/kg) of THP orally. The results showed that THP could decrease the blood glucose, TC, TG, LDL‐C levels, increase the body weight, HDL‐C, insulin levels, and enhance the activities of antioxidant enzyme system in alloxan‐induced diabetic mice. Furthermore, the histopathological examination of pancreas, liver, and kidney indicated that THP could protect and reverse β‐cells in diabetic mice with low damage to liver and kidney, which suggests that THP may stimulate pancreatic release of insulin and can be an effectively potential candidate for diabetes mellitus.     

Experimental Neospora caninum infection in chickens (Gallus gallus domesticus) with oocysts and tachyzoites of two recent isolates reveals resistance to infection

The importance of birds in the biological cycle of Neospora caninum is not clear. We report unsuccessful Neospora infection in chickens (Gallus gallus domesticus) using two isolates of N. caninum. In experiment #1, 30 White Leghorn chickens were orally inoculated with viable N. caninum oocysts (NC-SP1 isolate, 200 oocysts per bird) via the crop at 21 days of age. Groups of three birds were euthanised at intervals of 7 days (a total of 9 weeks) and one group was challenged with the same oocyst dose at 37 days p.i. and observed for 11 weeks. Blood samples were collected weekly, and sera were tested using IFAT. Chicken tissues were collected for PCR, quantitative PCR and immunohistochemistry. Two dogs approximately 45 days of age were fed with tissues from chickens euthanised at 138 and 159 days p.i. The results indicated that the chickens were resistant to neosporosis as revealed by failure to seroconvert, to detect parasite DNA or N. caninum antigen by immunohistochemistry in inoculated bird tissues, and by no oocyst excretion by the dogs fed avian tissues. Similar results were obtained in experiment #2, in which 34 1-week-old chickens were each s.c. inoculated with 100,000 tachyzoites of the NcWTDMn1 isolate of N. caninum. The chickens were euthanised on days 7, 15, 22, 28, 36 and 60 p.i. At necropsy, all tissues and serum from each bird were collected. All chickens remained asymptomatic, and N. caninum antigen was not detected by immunohistochemistry. Seven chickens euthanised at day 60 p.i. demonstrated low (1:25 dilution) levels of antibodies by using the Neospora agglutination test. Two 12-week-old dogs fed tissues pooled from 10 inoculated chickens euthanised at day 60 p.i. did not excrete N. caninum oocysts. This investigation indicates that chickens are resistant to experimental infection by N. caninum.     

Reduced blood‐brain barrier expression of fatty acid‐binding protein 5 is associated with increased vulnerability of APP/PS1 mice to cognitive deficits from low omega‐3 fatty acid diets

Lower levels of the cognitively beneficial docosahexaenoic acid (DHA) are often observed in Alzheimer's disease (AD) brains. Brain DHA levels are regulated by the blood‐brain barrier (BBB) transport of plasma‐derived DHA, a process facilitated by fatty acid‐binding protein 5 (FABP5). This study reports a 42.1 ± 12.6% decrease in the BBB transport of ¹⁴C‐DHA in 8‐month‐old AD transgenic mice (APPswe,PSEN1?E9) relative to wild‐type mice, associated with a 34.5 ± 6.7% reduction in FABP5 expression in isolated brain capillaries of AD mice. Furthermore, short‐term spatial and recognition memory deficits were observed in AD mice on a 6‐month n‐3 fatty acid‐depleted diet, but not in AD mice on control diet. This intervention led to a dramatic reduction (41.5 ± 11.9%) of brain DHA levels in AD mice. This study demonstrates FABP5 deficiency and impaired DHA transport at the BBB are associated with increased vulnerability to cognitive deficits in mice fed an n‐3 fatty acid‐depleted diet, in line with our previous studies demonstrating a crucial role of FABP5 in BBB transport of DHA and cognitive function.

     

Extreme drought alters growth and interactions with exotic grasses,but not survival,for a California annual forb

Climate change will not only alter mean conditions, but increase the frequency and intensity of extreme events such as severe droughts. Yet the consequences of extreme drought for plant demography are poorly understood. We compared phenology, demographic rates and effects of competition with exotic grasses for the California annual forb Phacelia distans between a year with slightly below average precipitation (2011–2012) and one in extreme drought (2012–2013). We also contrasted these demographic responses with changes in seedling emergence rates and cover in the annual plant community. Early Phacelia mortality actually fell in the extreme drought year, as low October rainfall shifted germination to cooler conditions in November. Survival from mid December to flowering did not change between years. In contrast to expectations, competition with early-emerging exotic annual grasses did not reduce Phacelia spring survival in 2012–2013. A shorter window for fall germination that reduced priority effects may explain this result. Yet the 2012–2013 growing season ended a month earlier than in 2011–2012, significantly reducing Phacelia size at flowering and inflorescence production. Community seedling emergence and cover changed only weakly between the 2 years. Our results demonstrate the importance of within season precipitation patterns in determining whether a climatically extreme year will lead to extreme ecological outcomes. This work also illustrates how annual plant drought responses depend critically on germination behavior and phenology. An important future question is whether seed banks can sustain native forb population persistence through an extended drought with multiple years of low or failed reproduction.     

Effects of agar and agarose on VentTM DNA polymerase and Taq DNA polymerase

Summary We have studied the effects of agar and agarose on Vent DNA polymerase and Taq DNA polymerase. Agar strongly inhibited Vent DNA polymerase but only moderately inhibited Taq DNA polymerase. Such a difference may be due to the fact that the two polymerases belong to different structural families. When Vent DNA polymerase is used to amplify DNA from lambda plaques, agarose rather than agar is the solid medium of choice.     

The <Emphasis Type="Italic">Helicobacter pylori</Emphasis> cytotoxin CagA is essential for suppressing host heat shock protein expression

Bacterial infections typically elicit a strong Heat Shock Response (HSR) in host cells. However, the gastric pathogen Helicobacter pylori has the unique ability to repress this response, the mechanism of which has yet to be elucidated. This study sought to characterize the underlying mechanisms by which H. pylori down-modulates host HSP expression upon infection. Examination of isogenic mutant strains of H. pylori defective in components of the type IV secretion system (T4SS), identified the secretion substrate, CagA, to be essential for down-modulation of the HSPs HSPH1 (HSP105), HSPA1A (HSP72), and HSPD1 (HSP60) upon infection of the AGS gastric adenocarcinoma cell line. Ectopic expression of CagA by transient transfection was insufficient to repress HSP expression in AGS or HEK293T cells, suggesting that additional H. pylori factors are required for HSP repression. RT-qPCR analysis of HSP gene expression in AGS cells infected with wild-type H. pylori or isogenic cagA-deletion mutant found no significant change to account for reduced HSP levels. In summary, this study identified CagA to be an essential bacterial factor for H. pylori-mediated suppression of host HSP expression. The novel finding that HSPH1 is down-modulated by H. pylori further highlights the unique ability of H. pylori to repress the HSR within host cells. Elucidation of the mechanism by which H. pylori achieves HSP repression may prove to be beneficial in the identification of novel mechanisms to inhibit the HSR pathway and provide further insight into the interactions between H. pylori and the host gastric epithelium.     

Effects of primer-template mismatches on the polymerase chain reaction: human immunodeficiency virus type 1 model studies.

We investigated the effects of various primer-template mismatches on DNA amplification of an HIV-1 gag region by the polymerase chain reaction (PCR). Single internal mismatches had no significant effect on PCR product yield while those at the 3'-terminal base had varied effects. A:G, G:A, and C:C mismatches reduced overall PCR product yield about 100-fold, A:A mismatches about 20-fold. All other 3'-terminal mismatches were efficiently amplified, although the G:G mismatches appeared to be more sensitive to sequence context and dNTP concentrations than other mismatches. It should be noted that mismatches of T with either G, C, or T had a minimal effect on PCR product yield. Double mismatches within the last four bases of a primer-template duplex where one of the mismatches is at the 3' terminal nucleotide, in general, reduced PCR product yield dramatically. The presence of a mismatched T at the 3'-terminus, however, allowed significant amplification even when coupled with an adjacent mismatch. Furthermore, even two mismatched Ts at the 3'-terminus allowed efficient amplification.     

The nucleotide sequence of phenylalanine tRNA from Mycoplasma sp. (Kid)

The nucleotide sequence of Mycoplasma sp. (Kid) phenylalanine tRNA was determined to be pG-G-U-C-G-U-G-U-A-G-C-U-C-A-G-U-C-G-G-D-A-G-A-G-C-A-G-C- A-G-A-C-U-G-A-A-m(1)G-C-Psi-C-U-G-C-G-U-m(7)G-U-C-G-G-C-G-G-U-Psi-C-A-A-U-U-C-C-G-U-C-C-A-C-G-A-C-C-A-C-C-A(OH). It is characterized by the absence of ribothymidine and the presence of only few modified nucleotides.