鱼精多肽在抗氧化和抗疲劳方面的作用研究

鱼精多肽(Protamine hydrolysate)是由鱼精蛋白(Protamine)酶解作用后形成的多肽片段,含有较大比例的寡肽和氨基酸,具有很高的营养价值和多种生理功能。研究了其体外清除自由基的能力和体内抗疲劳及抗氧化方面的作用,结果发现鱼精多肽粗粉具有较强的羟自由基清除活性,也能明显提高Wistar大鼠的抗疲劳和抗氧化能力。     

好氧丰盛/缺氧饥饿模式驯化微生物积累内源聚合物并实现内源反硝化

为了探究好氧丰盛/缺氧饥饿（O/A-F/F）模式驯化的微生物的内源积累能力和缺氧段内源反硝化的特性,并比较其与传统好氧丰盛/饥饿（O-F/F）驯化模式的差别,以添加乙酸的生活污水为底物,在F/F=0.1的条件下,采用O/A-F/F和O-F/F两种不同模式驯化活性污泥.结果表明,O/A-F/F模式的PHA产率为0.52,O-F/F模式的PHA产率为0.78.虽然O/A-F/F模式的PHA产率低,但是O/A-F/F模式具有内源反硝化功能,比内源反硝化速率为2.76 mg/（h·g）.O/A-F/F模式下还原1 mol的硝酸盐需要2 mol的内源PHA.2种选择模式下的菌群结构呈现差异,O/A-F/F模式下优势菌群为Betaproteobacteria、Unclassified Bacteroidetes、Lgnavibacteria和Bacteroidia.O-F/F模式富集的微生物主要为Betaproteobacteria、Flavobacteria、Alphaproteobacteria和Gammaproteobacteria.O/A-F/F运行模式可以实现好氧阶段污水中有机物的内源储存并在缺氧段用于反硝化.

     

An ESI-MS/MS method for screening of small-molecule mixtures against glycogen synthase kinase-3beta (GSK-3beta)

Glycogen synthase kinase-3beta (GSK-3beta) is involved in the hyperphosphorylation of previously phosphorylated (primed) substrates, and is currently assayed using an approach based on the incorporation of gamma-(32)P-radiolabelled isotopes into substrate peptides. The requirement to detect hyperphosphorylation of a primed substrate poses a particular challenge for development of a high-throughput screening assay, as many current kinase assays are designed to produce a signal in the presence of any phosphorylation site, and thus are only suitable for beta-unphosphorylated substrates. Herein, we have developed an electrospray-ionization tandem mass spectrometry (ESI-MS/MS) assay to allow for direct detection of a hyperphosphorylated product which is formed in a solution reaction involving a primed peptide substrate (GSM peptide) and GSK-3beta. Optimum reaction conditions (level of Mg(2+), buffer type, ionic strength, pH, enzyme concentration, and reaction time) were established to both maintain the activity of GSK-3beta and allow for substrate and product quantification through ESI/MS/MS. We show that the MS-based assay allows for rapid determination of GSK-3beta activity from reaction volumes of approximately 40 microL and that it can be used to assess IC(50) values and the site of action of known inhibitors. It also can be used for automated screening of small-molecules mixtures to identify inhibitors of GSK-3beta.     

Seasonal changes in hepatocytic lipid droplets,glycogen deposits,and rough endoplasmic reticulum along the natural breeding cycle of female ohrid trout (Salmo letnica Kar.)—A semiquantitative ultrastructural study

This study on wild female Ohrid trout was primarily designed to provide a general overview of the breeding cycle influence upon selected aspects of hepatocytes. According with a semiquantitatively evaluation, some of these cell's structural compartments change during the breeding cycle. Structural modifications were disclosed in the relative occurrence of lipid, glycogen, and RER content during breeding cycle. The relative amount of lipid deposits in the hepatocytes was much greater in previtellogenesis, and decreased postspawning. So, while the seasonal changes in RER were positively related with the ovary maturation status, those of the lipid droplets followed an opposite trend. The hepatocytic glycogen occurred rarely, mainly in late‐vitellogenesis and spawning, suggesting that in this species such kind of energy storage is comparatively unimportant. Lipid accumulation and later usage is, probably, the relevant biochemical pathway for Ohrid trout in the wild. While glycogen and RER contents were positively correlated with the gonadosomatic index, lipids were negatively correlated. Additionally, glycogen inclusions were positively correlated with the plasma estradiol levels. When comparing seasonal patterns from wild Ohrid trout with those from well‐studied rainbow and brown trout (specimens studied were from aquaculture), there are contradicting results as to lipid and glycogen reserves, and also as to RER loads. The differences among the mentioned trout can result from intrinsic interspecies differences or may be associated with natural feeding conditions versus feeding with commercially prepared diets, or other factors. This study offers new data useful as standard to access liver pathology in wild and aquacultured Ohrid trout. Microsc. Res. Tech. 79:700–706, 2016. © 2016 Wiley Periodicals, Inc.     

不同NO3-浓度An/A/O-SBR系统PAOs-GAOs竞争及N2O释放特性

采用厌氧/缺氧/好氧运行的序批式生物反应器（An/A/O-SBR）,经不同NO₃^-浓度（10,20,30和40 mg/L,以氮计）长期驯化,考察了不同NO₃^-条件下An/A/O-SBR脱氮除磷及N₂O释放特性,基于不同微生物降解特性分析,确定了不同NO₃^-浓度下SBR系统内反硝化聚磷菌（denitrifying phosphorus accumulating organisms,DPAOs）和聚糖菌（glycogen-accumulating organisms, GAOs）竞争关系。结果表明：随NO₃^-浓度增加,总氮（TN）去除率由90%以上降至41.3%,TP去除率呈先增高后降低的趋势,N₂O产率（N₂O_emission/NO_x^-_removal）分别为1.68%、4.17%、8.92%和14.28%。An/A/O-SBR内微生物呈PAOs和GAOs共存的污染物降解特性,高浓度NO₃^-缺氧吸磷过程出现NO₂^-积累,抑制DPAOs活性,GAOs碳源竞争能力增强,NO₃^--N由10 mg/L增至40 mg/L,厌氧阶段PAOs的COD耗量比例由33.5%降至25.1%,相应GAOs的COD耗量由59.3%增至74.1%。DPAOs-GAOs共生体系内,反硝化过程NO₂^-/HNO₂积累耦合反硝化聚糖菌比例增加,加剧了高NO₃^-下An/A/O-SBR内N₂O释放。     

厌氧-限氧SBR处理低C/N生活污水SNDPR启动及N2O释放

以厌氧-限氧方式运行序批式生物反应器（SBR）,采用逐步降低进水碳氮比（C/N）方式驯化聚磷菌（PAOs）和聚糖菌（GAOs）,启动了低C/N生活污水同步脱氮除磷过程（SNDPR）,并考察了SNDPR内PAOs、GAOs间竞争关系及系统脱氮除磷性能过程N₂O释放特性。结果表明,C/N=7.0,SBR限氧段脱氮和除磷效率分别为83.5%和90%以上,N₂O产量为0.54 mg/L;C/N=3.0~3.5,脱氮和除磷效率分别降至60.1%和80.5%,N₂O产量达1.09 mg/L。SBR内不同反应阶段内源物质变化均表现出PAOs-GAOs共存特性。高C/N有利于微生物合成聚-β-羟基烷酸酯（PHA）并促进N₂O还原。C/N降低,SBR内污泥内源物质转化倾向于富集GAOs的降解特性。氨氧化菌（AOB）好氧反硝化过程及GAOs以PHA作为电子供体的内源反硝化过程促进了N₂O的释放。随C/N降低,SBR内污泥平均胞外聚合物（EPS）由43.4 mg/g VSS增至50.5 mg/g VSS,污泥容积指数（SVI）由99 ml/g增至127 ml/g。疏松型EPS（LB-EPS）内,蛋白质（PN）与多糖（PS）之比（PN/PS）随C/N增加而降低,污泥亲水性增加,不利于污泥脱水。     

不同温度及厌氧/好氧运行条件对聚磷菌衰减特性的影响

以富含90％±2％纯度聚磷菌(PAOs)的强化生物除磷系统(EBPR)为研究对象,考察了10℃厌氧、10℃好氧、20℃厌氧、20℃好氧4种运行条件下PAOs的衰减特征.结果表明:温度越高对应衰减速率越快,4个系统在1～9 d里衰减速率的平均值分别为:10℃厌氧:0.053/d; 10℃好氧:0.050/d;20℃厌氧:0.072/d;20℃好氧:0.145/d.其中4个系统由于细胞死亡引起的活性衰减速率分别为:10℃厌氧:0.019/d;10℃好氧:0.017/d;20℃厌氧:0.019/d;20℃好氧:0.03/d,占总活性衰减的比例分别为:35.8％、34％、26.4％、20.7％.在9d饥饿衰减期间,污泥中所含PHA与糖原的量总体呈下降趋势.相同温度下,糖原在厌氧衰减过程中降解速率大于好氧;在同样的厌氧、好氧衰减条件下,温度越高糖原降解速率越快.     

Monitoring of Brewing Yeast Propagation Under Aerobic and Anaerobic Conditions Employing Flow Cytometry

The vitality and viability of industrial strains of Saccharomyces cerevisiae was monitored during pilot plant experiments simulating yeast propagation under aerobic and anaerobic conditions. Industrial wort of 12°P original gravity was used as a growth substrate for yeast propagation. The work was carried out with three widely used Czech lager yeast industrial strains: strains 2, 7 and 95. Cell cycle, cell size, granularity, glycogen content, DNA and protein content were analyzed by flow cytometry. Significantly higher specific growth rates, higher content of yeast glycogen, earlier G2/M phase cells maximum, and faster cell protein creation was observed under aerobic conditions compared to anaerobic. Strains 7 and 95 showed losses in flocculation ability after aerobic propagation compared to anaerobic propagation. Under either aerobic or strictly anaerobic conditions, only strain 2 did not show a significant loss in flocculation ability.     

Aspects of transition cow metabolomics—Part II: Histomorphologic changes in the liver parenchyma throughout the transition period,in cows with different liver metabotypes and effects of a metaphylactic butaphosphan and cyanocobalamin treatment

The aims of this study were to evaluate histopathologic changes during the transition period, describe the histopathological features of the metabotypes identified in Part I (Schären et al., 2021b), and investigate effects of a metaphylactic treatment with butaphosphan and cyanocobalamin (BCC) on the liver parenchyma. Eighty German Holstein cows (mean 305-d production: 10,957 kg, range: 6,480–15,193 kg; mean lactation number: 3.9, range: 2–9) from a commercial dairy farm in Saxony, Germany, were enrolled in a randomized, prospective, triple-blinded study. Two groups received a treatment with BCC (5 or 10 mL/100 kg of body weight 10% butaphosphan and 0.005% cyanocobalamin, Catosal, Bayer Animal Health, n = 20 each) and one group a placebo treatment (NaCl 0.9%, n = 40). Liver biopsy specimens were collected 14 d antepartum (AP) and 7, 28, and 42 d postpartum (PP), routinely processed for histologic examination, and stained with hematoxylin and eosin, Sudan III, periodic acid-Schiff, and picrosirius red stains. The sections were assessed for fat and glycogen content and degenerative, inflammatory, fibrotic, and proliferative changes. The statistical analysis included the effects of the sampling day, the lactation number, the treatment, and the metabotype (A = medium, B = minor, C = large alterations in the liver metabolome profile between AP and PP status). There was mild to moderate fat infiltration in the liver of 37% of cows in the last 2 wk AP, and moderate to severe fat infiltration in 66% of cows in the first days PP. The degree of fat infiltration increased from 2 wk AP until the end of the first week PP, and then decreased until the end of the study period, at which time about 25% of cows had moderate to severe fatty infiltration. Lipidosis was positively correlated with the severity of liver cell degeneration, and negatively correlated with the degree of glycogen deposits. Complete glycogen depletion of hepatocytes was not observed in cows, even in the presence of severe hepatic lipidosis. Moderate to severe lymphocytic hepatitis was seen in 39% of cows throughout the study period, and cows with lactation numbers 5 or greater had perisinusoidal fibrosis more often than younger cows. Severe fibrosis and cirrhosis of the liver did not occur. Metabotype B animals exhibited a higher chance of fatty infiltration, lower glycogen storage, and perisinusoidal fibrosis and for this metabotype positive correlations were calculated between increased fat deposition in the liver and marked glycogen depletion, and increased degenerative, inflammatory, fibrotic, and proliferative changes of hepatic tissue. For the treatment with BCC, no significant effect was observed. In summary, during the transition period, the liver of dairy cows is characterized by fat accumulation and glycogen depletion and histologic signs of hepatitis and hepatocyte degeneration. These histomorphologic changes were accentuated in animals exhibiting little alterations in their liver metabolome profile across the transition period (metabotype B) and support the assumption of a decreased grass silage quality as a causative factor.