发酵产柴胡皂苷酶的研究

研究了用酶水解柴胡皂苷分子上的部分糖基，使皂苷部分水解生成低糖、高活性皂苷的方法．探讨了3种菌株对柴胡皂苷糖基的水解能力，得到1种具有较高活性的菌株SP．以2，用于水解柴胡皂苷.通过TLC法分析得到柴胡皂苷酶解最佳反应条件为40℃、24h,pH=5．0，在此基础上的酶反应回收率为118.8％.     

玉米/蒜苗套作系统中土壤微生物和土壤酶状况分析

对玉米蒜苗套作根区进行隔膜、隔网和无隔处理以及各自单作处理,通过处理间的相互比较,分析影响套作产量的主要土壤因素。结果表明:套作蒜苗土壤微生物上升其主要原因是根系间的非接触物质交换;套作玉米除此外,地上部环境的改变也使根部土壤微生物数量上升。根系间的非接触效应还使蒜苗、玉米土壤过氧化氢酶、蔗糖酶活性提高,而环境因素的改变也使玉米过氧化氢酶,蒜苗脲酶活性提高。相关性分析表明:微生物对蒜苗根际土壤酶活性的影响大于对玉米土壤酶的影响。     

Elevation of Tear MMP-9 Concentration as a Biomarker of Inflammation in Ocular Pathology by Antibody Microarray Immunodetection Assays

Matrix metalloproteinases are a family of enzymes fundamental in inflammatory processes. Between them, MMP-9 is up-regulated during inflammation; thus, its quantification in non-invasive fluids is a promising approach for inflammation identification. To this goal, a biomarker quantification test was developed for ocular inflammation detection using anti-MMP-9 antibody microarrays (AbMAs). After validation with eight healthy control tear samples characterized by ELISA, 20 samples were tested from individuals diagnosed with ocular inflammation due to: cataracts, glaucoma, meibomian gland dysfunction, allergy, or dry eye. Concentration values of tear MMP-9 were obtained for each sample, and 12 patients surpassed the pathological threshold (30 ng/mL). A significant elevation of MMP-9 concentration in the tears of glaucoma patients compared with healthy controls was observed. In order to evaluate the diagnostic ability, an ROC curve analysis was performed using our data, determining the optimal threshold for the test at 33.6 ng/mL of tear MMP-9. In addition, a confusion matrix was applied, estimating sensitivity at 60%, specificity at 88%, and accuracy at 68%. In conclusion, we demonstrated that the AbMAs system allows the quantification of MMP-9 in pathologies that involve inflammation of the ocular surface.     

基于Au NPs-CeO2@PANI纳米复合材料固定化酶的葡萄糖生物传感器

制备了一种基于金纳米粒子(Au NPs)、氧化铈纳米颗粒(CeO2)和导电聚苯胺(PANI)的具有核壳结构的纳米复合材料(Au NPs-CeO2@PANI),利用该纳米复合材料和壳聚糖形成的复合膜成功实现了对葡萄糖氧化酶(GOD)的固定.采用透射电镜和X射线衍射对Au NPs-CeO2@PANI材料进行了表征.电化学方法研究了传感器性能,结果表明基于Au NPs-CeO2@PANI纳米复合材料修饰的葡萄糖生物传感器线性范围为6.2×10-6 mol/L～2.8×10-3 mol/L,响应时间为5 s,检测下限为1.0×10-6 mol/L;相同条件下Au NPs-CeO2@PANI纳米复合材料修饰的电极也显示出了比单一或二者复合的纳米材料修饰电极更优越的性能.     

氮掺杂中空碳球氧化物模拟酶性能研究

纳米酶由于其独特、高效、稳定的催化性质而在生化反应中备受关注.本研究以碳酸钙微球为绿色模板剂,多巴胺为氮源与碳源,合成了氮掺杂中空碳球(N-HCSs).以3,3',5,5'-四甲基联苯胺(TMB)为底物,采用紫外分光光度法探究了 N-HCSs的类氧化物酶的催化活性,并研究其催化机理.结果表明,N-HCSs具有氧化物模拟...     

Fine Mapping and Functional Analysis of Major Regulatory Genes of Soluble Solids Content in Wax Gourd (Benincasa hispida)

Soluble solids content (SSC) is an important quality trait of wax gourd, but reports about its regulatory genes are scarce. In this study, the SSC regulatory gene BhSSC2.1 in wax gourd was mined via quantitative trait locus (QTL) mapping based on high-density genetic mapping containing 12 linkage groups (LG) and bulked segregant analysis (BSA)-seq. QTL mapping and BSA-seq revealed for the first time that the SSC QTL (107.658–108.176 cM) of wax gourd was on Chr2 (LG2). The interpretable phenotypic variation rate and maximum LOD were 16.033% and 6.454, respectively. The QTL interval contained 13 genes. Real-time fluorescence quantitative expression analysis, functional annotation, and sequence analysis suggested that Bch02G016960, named BhSSC2.1, was a candidate regulatory gene of the SSC in wax gourd. Functional annotation of this gene showed that it codes for a NADP-dependent malic enzyme. According to BhSSC2.1 sequence variation, an InDel marker was developed for molecular marker-assisted breeding of wax gourd. This study will lay the foundation for future studies regarding breeding and understanding genetic mechanisms of wax gourd.     

Establishing In-House Cutoffs of CSF Alzheimer’s Disease Biomarkers for the AT(N) Stratification of the Alzheimer Center Barcelona Cohort

Background: Clinical diagnosis of Alzheimer’s disease (AD) increasingly incorporates CSF biomarkers. However, due to the intrinsic variability of the immunodetection techniques used to measure these biomarkers, establishing in-house cutoffs defining the positivity/negativity of CSF biomarkers is recommended. However, the cutoffs currently published are usually reported by using cross-sectional datasets, not providing evidence about its intrinsic prognostic value when applied to real-world memory clinic cases. Methods: We quantified CSF Aβ1-42, Aβ1-40, t-Tau, and p181Tau with standard INNOTEST^® ELISA and Lumipulse G^® chemiluminescence enzyme immunoassay (CLEIA) performed on the automated Lumipulse G600II. Determination of cutoffs included patients clinically diagnosed with probable Alzheimer’s disease (AD, n = 37) and subjective cognitive decline subjects (SCD, n = 45), cognitively stable for 3 years and with no evidence of brain amyloidosis in 18F-Florbetaben-labeled positron emission tomography (FBB-PET). To compare both methods, a subset of samples for Aβ1-42 (n = 519), t-Tau (n = 399), p181Tau (n = 77), and Aβ1-40 (n = 44) was analyzed. Kappa agreement of single biomarkers and Aβ1-42/Aβ1-40 was evaluated in an independent group of mild cognitive impairment (MCI) and dementia patients (n = 68). Next, established cutoffs were applied to a large real-world cohort of MCI subjects with follow-up data available (n = 647). Results: Cutoff values of Aβ1-42 and t-Tau were higher for CLEIA than for ELISA and similar for p181Tau. Spearman coefficients ranged between 0.81 for Aβ1-40 and 0.96 for p181TAU. Passing–Bablok analysis showed a systematic and proportional difference for all biomarkers but only systematic for Aβ1-40. Bland–Altman analysis showed an average difference between methods in favor of CLEIA. Kappa agreement for single biomarkers was good but lower for the Aβ1-42/Aβ1-40 ratio. Using the calculated cutoffs, we were able to stratify MCI subjects into four AT(N) categories. Kaplan–Meier analyses of AT(N) categories demonstrated gradual and differential dementia conversion rates (p = 9.815⁻²⁷). Multivariate Cox proportional hazard models corroborated these findings, demonstrating that the proposed AT(N) classifier has prognostic value. AT(N) categories are only modestly influenced by other known factors associated with disease progression. Conclusions: We established CLEIA and ELISA internal cutoffs to discriminate AD patients from amyloid-negative SCD individuals. The results obtained by both methods are not interchangeable but show good agreement. CLEIA is a good and faster alternative to manual ELISA for providing AT(N) classification of our patients. AT(N) categories have an impact on disease progression. AT(N) classifiers increase the certainty of the MCI prognosis, which can be instrumental in managing real-world MCI subjects.     

一种验证SPR传感器金膜表面固定蛋白质分子有效性的简易途径

利用表面等离子共振传感器进行免疫检测的实验中,需要在化学性质稳定的金膜表面固定蛋白质分子探针。如何验证一种方法是否能够成功固定蛋白质分子具有非常重要的现实意义。提出的验证方法采用酶反应作为标志,如果某种方法确实能有效固定蛋白质分子,那么它也能固定酶。将固定好酶的金膜浸入相应的底物溶液中发生变色反应,测量底物溶液的吸收光谱,即可根据结果判断是否有效固定了酶,同时达到判断这种方法是否可固定蛋白质的目的。具有快速、方便、低成本等优点。采用经过验证的方法固定抗IgG后通入IgG,实验响应良好。     

THE MEASUREMENT OF DIMETHYL SULPHOXIDE IN BARLEY AND MALT AND ITS REDUCTION TO DIMETHYL SULPHIDE BY YEAST

Dimethyl sulphoxide (DMSO) is a normal component of malt and barley. A method is described for its extraction and estimation. DMSO is produced by the oxidation of dimethyl sulphide (DMS), particularly during kilning of malt, and higher levels are found in malts subjected to ale kilning schedules. DMS may also be oxidized during wort preparation. DMSO can be reduced to DMS by yeast in glucose/salts medium, by yeast cell suspensions and by a cell-free extract. Reduction of DMSO is inhibited by methionine sulphoxide. The results suggest that reduction of DMSO may account for the DMS produced during fermentation of ale and lager worts.