Effect of yeast with bacteriocin from rumen bacteria on laying performance,blood biochemistry,faecal microbiota and egg quality of laying hens

The purpose of this study was to evaluate the effect of yeast with bacteriocin from Ruminococcus albus 7 (albusin B) on physiological state and production performance of laying hens. One hundred and twenty 26‐week‐old Single Comb White Leghorn (Hyline) laying hens were assigned into five groups including: (i) control group, (ii) yeast control (YC), (iii) 0.125% yeast with bacteriocin (0.125B), (iv) 0.25% yeast with bacteriocin (0.25B) and (v) 0.5% yeast with bacteriocin (0.5B). All supplements were added to the experimental diets of the hens from 26 to 46 weeks of age. Samples were collected every 4 weeks. Blood samples were collected from the wing vein for blood biochemical parameters assay, and faecal samples were collected by swab for the microbiota test. The egg production performance was recorded daily, and fresh eggs were collected for quality test. The blood biochemical assay results indicated that the addition of yeast with bacteriocin decreased the AST (aspartate aminotransferase) activity and it also affects the lactate concentration in laying hen blood. The result of egg quality indicated that yeast with bacteriocin supplementation had no effect on the mass of yolk and the strength of eggshell, but it had positive effect on the laying performance under hot environment. Low concentration bacteriocin (0.125B) supplementation could decrease total yolk cholesterol. The faecal microbiota result indicated that the supplementation of bacteriocin increased the lactobacilli counts. The yeast with bacteriocin supplementation significantly decreased the clostridia counts under hot environment condition, especially in hens receiving 0.25B. Combining the data from clinic chemistry, faecal microbiota, egg production and egg quality, the 0.25B supplementation may result in the best physiological parameter and egg production performance of laying hen.     

Temporal distribution and pathogenicity of the predominant tomato‐infecting begomoviruses in Taiwan

Between 1998 and 2009, the four tomato‐infecting begomovirus species detected in Taiwan were Ageratum yellow vein Hualien virus (AYVHuV), Tomato leaf curl Taiwan virus (ToLCTWV), Tomato yellow leaf curl Thailand virus (TYLCTHV) and a newly defined species Tomato leaf curl Hsinchu virus (ToLCHsV). AYVHuV was detected occasionally in 2003 and ToLCHsV only in 2000–2001, whilst ToLCTWV was detected throughout the period. TYLCTHV was first detected in 2005. Between 1998 and 2005, >99% of the begomovirus‐positive samples were infected with ToLCTWV. In 2007 in western Taiwan, 16% of the positive samples were infected with ToLCTWV, 35% with TYLCTHV and 49% with mixed infection (ToLCTWV/TYLCTHV). In contrast, in eastern Taiwan the proportions were 84% ToLCTWV, 2% TYLCTHV and 14% mixed infection. However, throughout Taiwan in 2008–2009, most positive samples were either identified as TYLCTHV (51%) or mixed infection (ToLCTWV/TYLCTHV; 41%), and only 8% were ToLCTWV. This shows a clear trend of shifting from ToLCTWV to TYLCTHV and mixed infection over a short time period in Taiwan. Sequence analyses indicated that tomato‐infecting AYVHuV, an apparent recombinant between ToLCTWV and AYVHuV from Ageratum, represents a new strain Hsinchu. TYLCTHV Taiwan isolates were highly similar to each other, whereas ToLCTWV isolates had greater diversity and were classified into three strains which had one country‐wide and two local distributions. ToLCTWV and TYLCTHV were confirmed as monopartite and bipartite begomoviruses, respectively, by agroinfection followed by transmission with Bemisia tabaci biotype B. In addition, TYLCTHV was found to be mechanically transmissible together with viral DNA‐B.     

In vitro inhibition of the activation of Pro-matrix Metalloproteinase 1 (Pro-MMP-1) and Pro-matrix metalloproteinase 9 (Pro-MMP-9) by rice and soybean Bowman-Birk inhibitors

The in vitro inhibitory activity of the rice Bowman-Birk inhibitor (rBBI) or soybean Bowman-Birk inhibitor (sBBI) against trypsin-catalyzed activation of pro-matrix metalloproteinase 1 or 9 (pro-MMP-1 or pro-MMP-9), respectively, was investigated using electrophoresis with silver staining, heparin-enhanced zymography, biotinylated gelatin, Biotrak assay, and fluorescence quenched substrate hydrolysis. rBBI at concentrations of 0.08-0.352 mg/mL dose-dependently inhibited the in vitro activation of 45 microg/mL pro-MMP-1 by trypsin. Heparin-enhanced zymography analysis of pro-MMP-1, trypsin-activated MMP-1, and a mixture of pro-MMP-1-trypsin-rBBI showed clear zones associated with trypsin-activated MMP-1 and the absence of clear zones in lanes containing pro-MMP-1 or a mixture of pro-MMP-1, trypsin, and rBBI. The results of the Biotrak assay also indicated that rBBI dose-dependently suppressed the activation of pro-MMP-1 by trypsin. sBBI dose-dependently inhibited the activation of 100 microg/mL of pro-MMP-9 by trypsin. Biotinylated gelatin assays demonstrated that pro-MMP-9 or pro-MMP-9 in the presence of trypsin and BBI did not hydrolyze gelatin, whereas p-aminophenylmercury acetate (APMA)-activated MMP-9 and trypsin-activated MMP-9 caused significant hydrolysis of gelatin. Quenched fluorescence substrate hydrolysis for total MMP activity showed that pro-MMP-1 or pro-MMP-9 did not hydrolyze the substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2; active MMP-1 or MMP-9 hydrolyzed the substrate, but lower substrate hydrolysis was obtained when pro-MMP-1 or pro-MMP-9 was incubated with trypsin in the presence of increasing concentrations of rBBI. The results are discussed in light of the role of MMP-1 and MMP-9 in the process of angiogenesis and the potential of rBBI or sBBI as a functional food ingredient.     

2,2'-Diphenyl-1-picrylhydrazyl radical-scavenging active components from adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) hulls

An activity-directed fractionation and purification process was used to identify the antioxidative components of adlay hulls. Hulls of adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) were extracted with methanol and then separated into water, 1-butanol, ethyl acetate, and hexane fractions. The 1-butanol-soluble fraction exhibited greater capacity to scavenge 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radicals when compared with fractions soluble in water, ethyl acetate, and hexane phases. The 1-butanol fraction was then subjected to separation and purification using Diaion HP-20 chromatography, silica gel chromatography, and HPLC. Six compounds showing strong antioxidant activity were identified by spectroscopic methods ((1)H NMR, (13)C NMR, IR, and MS) and by comparison with authentic samples to be coniferyl alcohol (1), syringic acid (2), ferulic acid (3), syringaresinol (4), 4-ketopinoresinol (5), and a new lignan, mayuenolide (6).     

Ursolic acid induces allograft inflammatory factor-1 expression via a nitric oxide-related mechanism and increases neovascularization

Ursolic acid (UA), a triterpenoid compound found in plants, is used in the human diet and in medicinal herbs and possesses a wide range of biological benefits including antioxidative, anti-inflammatory, and anticarcinogenic effects. Endothelial expression of allograft inflammatory factor-1 (AIF-1) mediates vasculogenesis, and nitric oxide (NO) produced by endothelial NO (eNOS) represents a mechanism of vascular protection. It is unclear whether UA affects the neovascularization mediated by AIF-1 and eNOS expression. This study investigated the effects and mechanisms of UA on angiogenesis in vivo in hind limb ischemic animal models and in vitro in human coronary artery endothelial cells (HCECs). This study explored the impact of UA on endothelial cell (EC) activities in vitro in HCECs, vascular neovasculogenesis in vivo in a mouse hind limb ischemia model, and the possible role of AIF-1 in vasculogenesis. The results demonstrate that UA enhances collateral blood flow recovery through induction of neovascularization in a hind limb ischemia mouse model. In vitro data showed that UA increases tube formation and migration capacities in human endothelial cells, and exposing HCECs to UA increased AIF-1 expression through a NO-related mechanism. Moreover, UA administration increased capillary density and eNOS and AIF-1 expression in ischemic muscle. These findings suggest that UA may be a potential therapeutic agent in the induction of neovascularization and provide a novel mechanistic insight into the potential effects of UA on ischemic vascular diseases.     

Original Study: Comparison of three intraosseous access techniques in cats

Objective – To compare the placement feasibility and amount of bone trauma induced by 3 intraosseous (IO) access techniques in cats: an automatic impact penetration device (A), an automatic rotary insertion device (B), and a manual IO needle (C). Design – Prospective ex vivo study. Setting – University. Animals – Eighteen adult mixed breed feline cadavers. Interventions – Cadavers provided 72 total IO insertion locations divided equally between the right and left humerus and tibia. The 3 IO techniques were randomly allocated to these locations. Time to successful insertion, ease of insertion, and success rate were recorded. Each insertion site was analyzed for the number of bone fragments and defect diameter by computed tomography. Measurements and Main Results – Device B had lower time of insertion (P=0.01) compared with devices A and C. Device B had better ease of insertion scores (P<0.01) compared with devices A and C. No differences were detected between insertion sites (tibia versus humerus). No differences in the number of bone fragments, defect diameter, or success rate were detected among devices (P=0.06, 0.31, and 0.14, respectively). Conclusions – All 3 IO access methods evaluated yield acceptable results. Device B is significantly faster and easier to place in cat cadavers when compared with other methods.     

Evaluation of an indirect oscillometric blood pressure monitor in normotensive and hypotensive anesthetized dogs

Objective – To determine the accuracy and precision of an oscillometric noninvasive blood pressure device as a predictor of invasive direct blood pressure in healthy anesthetized hypotensive and normotensive dogs. Design – Prospective observational study. Setting – University teaching hospital. Animals – Eight crossbred adult dogs. Interventions – Anesthesia was induced with propofol and maintained with isoflurane. A catheter was placed in the dorsal pedal artery to record systolic, mean, and diastolic arterial blood pressures (aSAP, aMAP, and aDAP, respectively). The noninvasive blood pressure device cuff was placed around the contralateral front limb to record noninvasive systolic, mean, and diastolic blood pressure (nSAP, nMAP, and nDAP). Two states of blood pressure (BP) were studied: baseline state was established by keeping end‐tidal isoflurane concentration at 1.2±0.1%. The hypotensive state was achieved by maintaining the same isoflurane concentration while withdrawing approximately 40% of the animal's blood volume until aMAP was stable at approximately 40 mm Hg. At the end of the study, blood was returned to the animal and it was allowed to recover from anesthesia. Measurements and Main Results – Agreement between the direct and indirect BP measurements was determined by the Bland‐Altman method. The SAP and MAP but not DAP bias varied significantly between each BP state. Normotensive absolute biases (mean [SD]) for SAP, MAP, and DAP were ?14.7 mm Hg (15.5 mm Hg), ?16.4 mm Hg (12.1 mm Hg), and ?14.1 mm Hg (15.8 mm Hg), respectively. Absolute biases during the hypotensive state for SAP, MAP, and DAP were ?32 mm Hg (22.6 mm Hg), ?24.2 mm Hg (19.5 mm Hg), and ?16.8 mm Hg (17.2 mm Hg), respectively. Conclusion – The oscillometric device was not reliably predictive of intra‐arterial BP during hypotension associated with acute hemorrhage.     

大豆质核互作雄性不育系NJCMS2A及其保持系的花药蛋白质组比较研究

大豆质核互作雄性不育系NJCMS2A是以栽培大豆组合（N8855 ´ N1628）F₂不育株为母本,以N1628为父本,通过连续回交选育而成的,N1628（或NJCMS2B）为其同型保持系。对NJCMS2A和NJCMS2B的二胞花粉期花药进行蛋白质组比较分析,获得重复性好的双向电泳图谱,在分子量18.4~116.0 kD、等电点4~7线性范围内,检测到约217个蛋白点,其中差异表达蛋白点25个,包括在NJCMS2A 中出现而在NJCMS2B中缺失的蛋白点13个,在NJCMS2A中缺失而在NJCMS2B中出现的蛋白点10个,另有2个蛋白点的表达量在NJCMS2B中比在NJCMS2A中明显增强。利用基质辅助激光解吸飞行时间质谱（MALDI-TOF-MS）技术对差异表达蛋白进行分析,获得肽质量指纹图谱,用Mascot软件搜索NCBInr数据库,鉴定出14个差异表达蛋白,其中10个在NJCMS2A中出现而在NJCMS2B中缺失和4个在NJCMS2A中缺失而在NJCMS2B中出现。对热激蛋白22 kD、半胱氨酸蛋白酶、V型H+-ATP酶A亚基、MADS盒蛋白和淀粉分枝酶等主要差异蛋白进行功能分析,推测不育系NJCMS2A雄性不育性可能与能量代谢紊乱、细胞程序化死亡（PCD）、淀粉合成受抑制和花器官发育调节基因作用失控等有关。     

Chemoinhibitory effect of mulberry anthocyanins on melanoma metastasis involved in the Ras/PI3K pathway

Anthocyanins richly exist in mulberry plants and have been well characterized to have various bioactive properties. However, the antimetastasis properties of mulberry anthocyanins (MACs) remain unclear. The objectives of this study were to investigate the inhibitory effects of MACs on the metastasis of B16-F1 cells under noncytotoxic concentrations. Further investigation revealed that the antimetastatic effect of MACs was also evident in a C57BL/6 mice model. First, MACs exhibited an inhibitory effect on the migration ability by wound healing assay and Boyden chamber assay. In the cancer cell metastasis process, matrix degrading proteinases are required. B16-F1 cells treated with MACs at various concentrations showed reduced extracellular matrix (ECM) proteinases including matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) by gelatin zymography assay. The results of the Western blotting assay demonstrated that the expression levels of Ras, phosphoinositide 3-kinase (PI3K), phospho-Akt, and nuclear factor kappa B (NF-kappaB) in the MACs-treated B16-F1 cells were reduced. Therefore, it was suggested that MACs could mediate B16-F1 cell metastasis by reduction of MMP-2 and MMP-9 activities involving the suppression of the Ras/PI3K signaling pathway. Besides, B16-F1 melanoma cells were also injected into the right groin of the C57BL/6 mice, and the mice were fed with MACs at the same time. The hematoxylin-eosin stain (H&E stain) and immunohistochemistry stain showed that the MACs inhibited the mtastasis of B16-F1 cells in vivo. Taken together, the findings proved the inhibitory effect of MACs on the growth and metastasis of B16-F1 cells. These results indicated that MACs might be offered for future application as an antimetastatic agent.