Determination of Indoxacarb Residue in Foodstuffs of Plant and Animal Origin by GC-ECD and LC-MS/MS

An effective method for the trace analysis of indoxacarb residue in foodstuffs of plant and animal origin [grapefruit, ginger, fresh soybean, bamboo shoot, qing-gen-cai （cruciferous vegetable）, chicken, fish, and pork] was developed using gas chromatography （GC-ECD） and liquid chromatography tandem mass spectrometry （LC-MS/MS）. Samples were extracted using acetone and n-hexane mixed solvent （1：2, v/v） and then purified using solid-phase extraction （SPE） columns. The extracts were analyzed using GC-ECD and LC-MS/MS. The multiple reaction monitoring （MRM） scheme used involved transitions of the precursor ions to selected two product ions in which one pair for identification was m/z 529 → 293 and another pair for quantification was m/z 529 → 249. The detection limits （LODs） of the method were 0.0015 and 0.0006 mg kg^-1, and the quantification limits （LOQs） were 0.005 and 0.002 mg kg^-1 for GC-ECD and LC-MS/MS, respectively. The relative standard deviations （RSDs） of recovery for indoxacarb were lower than 15% in 10 types of agro-products. Ten repetitive determinations of recovery achieved good reproducibility for indoxacarb and the recovery ranged from 72.08 to 113.74%. The proposed procedure was applied to the analysis of several real samples of different origin from Fujian Province, China, and 299 samples were screened for indoxacarb residue, of which 5 positive samples were found.

Safety assessment of Bifidobacterium longum JDM301 based on complete genome sequences

AIM： To assess the safety of Bifidobacterium/ongum （B. longum） JDM301 based on complete genome sequences. METHODS： The complete genome sequences of JDM301 were determined using the GS 20 system. Putative virulence factors, putative antibiotic resis- tance genes and genes encoding enzymes respon- sible for harmful metabolites were identified by blast with virulence factors database, antibiotic resistance genes database and genes associated with harmful metabolites in previous reports. Minimum inhibitory concentration of 16 common antimicrobial agents was evaluated by E-test RESULTS： JDM301 was shown to contain 36 genes as- sociated with antibiotic resistance, 5 enzymes related to harmful metabolites and 162 nonspecific virulence fac- tors mainly associated with transcriptional regulation, adhesion, sugar and amino acid transport. B. longum JDM301 was intrinsically resistant to ciprofloxacin, ami- kacin, gentamicin and streptomycin and susceptible to vancomycin, amoxicillin, cephalothin, chloramphenicol, erythromycin, ampicillin, cefotaxime, rifampicin, imi- penem and trimethoprim-sulphamethoxazol. JDM301 was moderately resistant to bacitracin, while an earlier study showed that bifidobacteria were susceptible to this antibiotic. A tetracycline resistance gene with the risk of transfer was found in JDM301, which needs to be experimentally validated. CONCLUSION： The safety assessment of JDM301 using information derived from complete bacterial ge- nome will contribute to a wider and deeper insight into the safety of probiotic bacteria.

Evidence for the involvement of NOD2 in regulating colonic epithelial cell growth and survival

AIM: To investigate the function of NOD2 in colonic epithelial cells (CEC). METHODS: A combination of in vivo and in vitro analyses of epithelial cell turnover in the presence and absence of a functional NOD2 protein and, in response to enteric Salmonella typhimurium infection, were used. shRNA interference was also used to investigate the consequences of knocking down NOD2 gene expression on the growth and survival of colorectal carcinoma cell lines. RESULTS:In the colonic mucosa the highest levels of NOD2 expression were in proliferating crypt epithelial cells. Muramyl dipeptide (MDP), that is recognized by NOD2, promoted CEC growth in vitro . By contrast,the growth of NOD2-deficient CECs was impaired. In vivo CEC proliferation was also reduced and apoptosis increased in Nod2-/- mice, which were also evident following enteric Salmonella infection. Furthermore, neutralization of NOD2 mRNA expression in human colonic carcinoma cells by shRNA interference resulted in decreased survival due to increased levels of apoptosis. CONCLUSION: These findings are consistent with the involvement of NOD2 protein in promoting CEC growth and survival. Defects in proliferation by CECs in cases of CD may contribute to the underlying pathology of disrupted intestinal homeostasis and excessive inflammation.

Genetic Diversity and Genetic Structure of Maize Inbred Lines from Yunnan Revealed by SNP Chips

[Objectives]The genetic diversity and population genetic structure of 107 inbred lines of maize in Yunnan were analyzed,in order to provide technical support for maize germplasm innovation,genetic improvement of germplasm resources,variety management,and lay a solid foundation for exploring genes related to fine traits in the future.[Methods]The 107 maize inbred lines generalized in Yunnan were selected,and 45 backbone inbred lines commonly used in China were used as reference for heterotic group classification.On Axiom Maize 56K SNP Array platform,maize SNP chips(56K)were used to scan the whole maize genome,and the NJ-tree model of Treebest was used to construct a phylogenetic tree.Principal component analysis(PCA)was conducted by GCTA(genome-wide complex trait analysis)to reveal the genetic diversity and population genetic structure.[Results]In the 107 Yunnan local inbred lines,5533 uniformly distributed high-quality SNP marker sites were finally detected.Based on the analysis of these SNP marker sites,Nei s gene diversity index(H)of 107 maize germplasm genes was 0.2981-0.5000 with an average value being 0.4832,and polymorphism information content(PIC)values were 0.2536-0.3750 with an average value being 0.3662.The minimum allele frequency value was 0.5000-0.8178 with an average value being 0.5744.The analysis of population genetic structure showed that when K=6,the maximum value of△K was the maximum,which meant that the inbred lines used in this study could be divided into six groups.They were Tangsi Pingtou blood relationship group,PB blood relationship group,335 female blood relationship group,Zi 330 and the Lüda Honggu blood relationship group,unknown group 1 and unknown group 2.No inbred lines were divided into other heterotic groups.Among them,37 inbred lines from the 2 unknown groups could not be classified into the same group as the 10 known heterotic groups in China.The results of principal component analysis showed that the 107 maize inbred lines generalized in Yunnan could be clearly distinguished from the backbone maize inbred lines commonly used in China.Most of the maize inbred lines in Yunnan were concentrated near the reference backbone inbred lines.But some Yunnan inbred lines were far away from the reference inbred lines commonly used in China.[Conclusions]The maize germplasm resources in Yunnan area were rich in genetic diversity,including multiple heterotic groups,and there was a rich genetic basis of breeding parents.They could be clearly distinguished from the backbone inbred lines commonly used in China,and some of them had a long genetic distance from the backbone inbred lines.The resources which have good application potential can be used to create new heterotic groups.

Extraction and Powder Product of Fructo-oligosaccharide from Jerusalem Artichoke

Fructo-oligosaccharide （FOS） is a type of dietary fiber. It can not be digested by enzymes in the body. It also helps to improve a digestion in the intestine, an excretion system and prevent the cancer. Jerusalem artichoke is scanned and analyzed by High Performance Liquid Chromatography （HPLC） for quantity of FOS. They are selected for extraction and drying process. Studies on the extraction for a powder making, the conditions are optimum for maximum yield. Experiments manage using a factorial real 2 × 4 × 6 in 2 blocks of Randomized Complete Block Design （RCBD） models. The first factor studies on two types of solvent. The second factor is the four levels of temperature in the extraction with water at 25 ℃, 35 ℃, 50℃ and 60℃ for 30 min. The third factor is the ratio of sample per solvent as 5, 6, 10, 15, 20 and 30 times. The data and comparison of average is analyzed by Duncan＇s New Mutiple Range Test at the significant level 0.05. The concentrated extracts are processed to powder by freeze drying, a hot air and vacuum drying. Optimal conditions control the temperature and time by heating in both vacuum and conventional oven. The results of temperature on the extraction are significantly different at a = 0.05. In drying process with high temperature, the color of product is more yellow （b） and less white （L）. The difference of color value is statistically significant at level a = 0.05. The sensory evaluation of food products are added the extracted powder and attribute by panelist. High level acceptance of product is aspectual about texture and flavor. FOS is the trend of functional foods for health. It is focus on low energy and on obesity including increasing the absorption of calcium.

Fractal image analysis and bruise damage evaluation of impact damage in guava

Impact bruise damage and quality of‘Gim Ju’guava were investigated for different drop heights and number of drops using fractal image analysis.For the impact test,a stainless-steel metal ball(250 g)was dropped on fruit from three drop heights(0,0.3,0.6 m)either once or five times.Fruit quality was evaluated for impact energy,bruise area(BA),bruise volume(BV),bruise susceptibility,bruise score and pulp color(L*,a*,b*and C values).The fractal dimension(FD)value using fractal image analysis was analyzed at the bruise region.Results showed that five drops(0.3 m)with a high impact energy(3678.75 J)and a single drop(0.6 m)with a low impact energy(1471.50 J)exhibited no significant in BA,BV,bruise score as well as all color values(L*,a*,b*and C).While the FD value of a single drop from 0.6 m had a higher FD value than that of five drops from 0.3 m.It is indicated that FD exhibited a better performance to classify impact bruising level of guava than BA,BV and color parameters.The FD value gradually decreased with increase of storage time and bruise severity.The correlation coefficient(r)values of FD(r=−0.794 and−0.745)between BA and BV were more significant than those L*(r=−0.660 and−0.615)and a*(r=0.579 and 0.473).The coefficient of determination(R^(2))of the polynomial equation in bruised fruit(R^(2)=0.85 to 0.99)was greater than the control(no bruise)(R^(2)=0.80).A higher R^(2)val(0.88 and 0.92)was exhibited at five drops.Interestingly,FD analysis showed greater potential than color measurement to assess bruise impact damage in guava.

稳定性氢同位素分析在牛肉产地溯源中的应用

探讨了利用高温热解炉和同位素比率质谱仪(IRMS)联用测定肉品中稳定性氢同位素比率的方法,并利用此方法测定了我国不同地域来源脱脂牛肉、牛尾毛中氢同位素比率。分析了牛组织中氢同位素组成与地域经度、纬度及海拔高度变化的关系,以及稳定性氢同位素用于牛肉产地溯源的可行性。结果表明,不同地域来源牛组织中δ2H值的差异显著,其与当地饮水中氢同位素组成密切相关,而且有随着地理纬度增加而减小的趋势;牛尾毛与牛肉中δ2H值的相关性显著。稳定性氢同位素是用于牛肉产地溯源的一项很有潜力的指标,且牛肉和牛尾毛中的氢同位素组成均可反映牛来源地的信息。

十字花科蔬菜在低硒小鼠病毒性心肌损伤中的作用

目的探讨十字花科蔬菜成分Sulforaphane(SFN)对低硒病毒性心肌损伤是否具有保护作用。方法采用低硒和常硒病毒性心肌损伤小鼠模型,观察给予SFN后全血谷胱甘肽过氧化物酶(GSH鄄Px)活性和心肌病理改变。结果不同硒状态下,SFN保护组GSH鄄Px活性高于相应未保护组(P<0.001),光镜下心肌病变检出率SFN保护组低于相应未保护组(P<0.001),病变程度也明显减轻。结论SFN对不同硒状态下病毒性心肌损伤具有保护作用。

不同浓度沙门氏菌细胞生长分布规律研究

选用沙门氏菌SL1344在含20mg/kg甜菜碱和NaCl质量分数分别为0.0%、3.5%和5.0%的最低营养培养基(BMM)环境中分别对大量细胞、少量细胞进行研究,并对3.5%NaCl条件下单个细胞的生长进行研究。重点介绍进行少量细胞研究的一般理论和方法,以及单个细胞研究中单细胞生长流动槽成像系统的应用。结果发现,大量细胞生长分布具有决定性,单细胞生长分布具有随机性的特点,并通过不同的生长模型初步阐述其关系。

黑曲霉菌pyrG缺陷株的建立

运用紫外线照射致突变方法建立了黑曲霉菌ATCC 12 0 49,134 96和N40 2等 3种菌株的乳清酸核苷 5′ 磷酸脱羧酶基因 (pyrG)缺陷株。其中ATCC 134 96是一种蛋白酶缺陷株。含黑曲霉菌pyrG基因的重组质粒 pY 1.2可使它们发生转化 ,成为Pyr+ ,转化效率约为 8～ 40转化子 / μgDNA。这些

乳酸乳球菌质粒pAG712的特性研究

乳酸乳球菌质粒pAG712(15.5kb)赋予乳酸乳球菌细胞凝聚表型,它是以θ模式进行复制,复制蛋白与其它以θ模式进行复制的质粒的复制蛋白具有极高的同源性。复制区上的BglII(4.2kb)片段决定质粒可以在乳酸乳球菌中进行复制。在本研究中,将抗生素抗性基因插入到质粒pAG712中,给予它选择性的标记,在研究中发现凝聚基因(aggregationgene)的存在导致了乳酸乳球菌质粒pAG712的分离不稳定性。

基于ITS1 DNA序列分析的几种酵母菌的分子分类

采用ITS1序列分析的手段,对来自Dekkera/Brettanomyces/Eeniella的15株菌株进行了分子分类学的研究。研究结果支持4个Dekkera/Brettanomyces种类的确认:D.anomala/B.anomalus,D.bruxellensis/B.bruxellensis,D.custersiana和D.naardenensis,以及把E.nana合并于Brettanomyces属的建议。此外,研究也揭示了ITS1在酵母分子分类学中的价值。

Heterogeneity across the murine small and large intestine

The small and large intestine of the gastrointestinal tract(GIT) have evolved to have discrete functions with distinct anatomies and immune cell composition.The importance of these differences is underlined when considering that different pathogens have uniquely adapted to live in each region of the gut.Furthermore,different regions of the GIT are also associated with differences in susceptibility to diseases such as cancer and chronic inflammation.The large and small intestine,given their anatomical and functional differences,should be seen as two separate immunological sites.However,this distinction is often ignored with findings from one area of the GIT being inappropriately extrapolated to the other.Focussing largely on the murine small and large intestine,this review addresses the literature relating to the immunology and biology of the two sites,drawing comparisons between them and clarifying similarities and differences.We also highlight the gaps in our understanding and where further research is needed.

风险意识、社会信任和公众参与决策制定:关于新兴技术的问题

从50年代起,公众就愈来愈关注与新兴技术有关的各种风险。重要的是不要只把关于风险意识的概念具体化为解释性的文字而不把其他因素例如人们对技术特别是新兴技术的控诉考虑进去。诸如伦理观念、对科学机构、风险管理者和信息提供者的信任和不信任以及感到被社会排斥于风险管理事务之外的想法,都应谊并入理论模式中用来解释公众对新兴技术抗拒的演变情况。加强努力把公众包容在风险管理事务中看来是一项纠正有关被社会排斥的认识的最好办法,不过重要的是公众的参与情况要以群众对参与的接受程度以及参与过程的有效性进行衡量。

Omics Approaches to Identify PotentialBiomarkers of Inflammatory Diseases in the FocalAdhesion Complex

Inflammatory diseases such as inflammatory bowel disease(IBD) require recurrent invasive tests, including blood tests, radiology, and endoscopic evaluation both to diagnose and assess disease activity, and to determine optimal therapeutic strategies. Simple ‘bedside’ biomarkers could be used in all phases of patient management to avoid unnecessary investigation and guide further management. The focal adhesion complex(FAC) has been implicated in the pathogenesis of multiple inflammatory diseases, including IBD, rheumatoid arthritis, and multiple sclerosis. Utilizing omics technologies has proven to be an efficient approach to identify biomarkers from within the FAC in the field of cancer medicine. Predictive biomarkers are paving the way for the success of precision medicine for cancer patients, but inflammatory diseases have lagged behind in this respect. This review explores the current status of biomarker prediction for inflammatory diseases from within the FAC using omics technologies and highlights the benefits of future potential biomarker identification approaches.

Green Extraction of Bioactive Compounds from Microalgae

At present,there is an increasing demand for natural bioactive compounds able to provide health benefits when included and consumed in a functional food or in a nutraceutical.In this regard,microalgae are promising natural sources with great potential,not only considering that these organisms are largely underexplored,but also because microalgae can be produced at large scale and their chemical composition might be tuned to over-synthesize a particular target compound.The use of advanced sustainable extraction techniques to recover these bioactive compounds is a must nowadays.This work presents an overview on the use of compressed fluid-based extraction techniques to obtain bioactive compounds from microalgae that can be seen also as a first step towards its recovery at larger scale.When relevant,the description of the analytical procedure used to chemically characterize the bioactive compounds is also included.

Pressurized Liquid Extraction of Pigments from Chlamydomonas sp.and Chemical Characterization by HPLC–MS/MS

Pressurized liquid extraction using green solvents(ethanol and water)was applied to extract photosynthetic pigments from the microalgae Chlamydomonas sp.An experimental design is employed to determine the effect of the extraction conditions on the extraction yield and the antioxidant capacity of the different extracts determined using 1,1-diphenyl-2-picryl-hydrazyl and TEAC tests.All the extracts show significant antioxidant activity in all assays.Besides,in this work,the carotenoid and chlorophyll composition of the Chlamydomonas extracts was analyzed by HPLC–DAD–APCI–MS/MS for the first time,being the main compounds lutein and pheophythin a′.

Carrageenan oligosaccharides and associated carrageenan-degrading bacteria induce intestinal inflammation in germ-free mice

Carrageenans(CGNs)are widely used in foods and pharmaceuticals although their safety remains controversial.To investigate the effects of CGNs and CGN-degrading bacteria in the human colon,we screened for CGN degradation by human fecal microbiota,and for inflammatory response to CGNs and/or CGN-degrading bacteria in germ free mice.Thin-layer chromatography indicated that high molecular weight(MW)CGNs(!100 kDa)remained undegraded in the presence of human fecal microbiota,whereas low MW CGNs,i.e.,k-carrageenan oligosaccharides(KCO,~4.5 kDa)were degraded when exposed to seven of eight human fecal samples,although sulfate groups were not removed during degradation.Bacteroides xylanisolvens and Escherichia coli isolates from fecal samples apparently degraded KCO synergistically,with B.xylanisolvens serving as the primary degrader.Combined treatment of KCO with KCO-degrading bacteria led to greater pro-inflammatory effects in the colon and rectum of germ-free mice than either KCO or bacteria alone.Similarly,p-p38-,CD3-,and CD79a-positive immune cells were more abundant in combined treatment group mice than in either single treatment group.Our study shows that KCO-degrading bacteria and the low MW products of KCO can promote proinflammatory effects in mice,and represent two key markers for evaluating CGN safety in foods or medicines.

Ileocecal lavage fluid for gut microbiota investigation in children with gastrointestinal diseases

Introduction The ileocecal region is the end point of colonoscopy and comprises the junction of the ileum,cecum,and appendix.The ileocecal region has been reported as a common lesion site for various intestinal diseases(e.g.Crohn’s disease[1],intestinal tuberculosis[2],and primary colorectal lymphoma[3]).However,to date,there have been few studies on the intestinal microbiota of the ileocecal region,because the sampling is relatively difficult and with only a local biopsy it is difficult to represent the whole region without deviation.Compared with ileocecal mucosa sampling,the collection of ileocecal lavage fluid is easier to be accepted by patients and can avoid the interference of human genomic DNA in the process of bacterial genome sequencing.