Paucity of the Sau3AI recognition sequence (GATC) in the genome of Methanococcus voltae

Summary High molecular weight genomic DNA isolated from the archaebacterium Methanococcus voltae by alkaline-SDS lysis was not effectively digested with the restriction enzyme Sau3AI, which recognizes the base sequence GATC. Mc. voltae DNA was also resistant to digestion by MboI and BamHI which recognize sites containing the same GATC sequence. Examination of a Mc. voltae genomic library prepared in Escherichia coli JM83 with a pUC vector revealed that the 5–10 kb inserts were still resistant to Sau3AI digestion, indicating a likely lack of the GATC sequence in Mc. voltae DNA.     

大鼠SR-BI胞外域与ApoA-Ⅰ蛋白相互作用的确认

先以含全长SR-BI cDNA序列的重组质粒pMD18-T-rS为摸板进行PCR反应扩增SR-BI得到胞外域cDNA片段,经测序证明正确后,定向克隆到酵母双杂交表达载体,然后与pGBKT7-ApoA-Ⅰ质粒共转化酵母细胞,通过报告基因及酵母交配试验确认了SR-BI的胞外域部分和ApoA-Ⅰ之间的确存,观察到ApoA-Ⅰ与SR-BI胞外域间的相互作用力比与全长的SR-BI问的相互作用力提高了10％。     

禽流感病毒实验检测研究进展

禽流感病毒(avian influenza viruses,AIV)给人类健康已带来严重威胁,而实验室快速、准确的诊断技术对禽流感的预防、控制及应急反应决策起着极其关键的作用,这使其成为了研究热点并取得了巨大进步。就禽流感的实验检测技术研究进展从病毒分离、免疫学诊断及分子诊断3个方面加以综述。     

血清载脂蛋白AI、鳞状细胞癌抗原与子宫内膜癌病理特征的关系及对淋巴结转移的预测研究

摘要 目的：分析血清载脂蛋白AI、鳞状细胞癌抗原与子宫内膜癌病理特征的关系及对淋巴结转移的预测价值。方法：选择我院自2020年7月至2022年7月收治的156例行手术治疗的子宫内膜癌患者作为研究对象,检测血清载脂蛋白AI、鳞状细胞癌抗原表达水平,分析不同临床分期、病理分级、子宫肌层浸润程度的子宫内膜癌血清载脂蛋白AI、鳞状细胞癌抗原表达水平的差异性,比较淋巴结转移组与非淋巴结转移组血清载脂蛋白AI、鳞状细胞癌抗原表达水平,通过受试者工作特征曲线（ROC）下面积（AUC）评价血清载脂蛋白AI联合鳞状细胞癌抗原对子宫内膜癌患者发生淋巴结转移的预测价值。结果：Ⅲ～Ⅳ期组血清载脂蛋白AI低于Ⅰ～Ⅱ期组,鳞状细胞癌抗原表达水平高于Ⅰ～Ⅱ期组（P＜0.05）;中低分化组血清载脂蛋白AI水平低于高分化组,鳞状细胞癌抗原表达水平高于高分化组（P＜0.05）;子宫肌层浸润≥1/2组血清载脂蛋白AI水平低于子宫肌层浸润＜1/2组,鳞状细胞癌抗原表达水平高于子宫肌层浸润＜1/2组（P＜0.05）;在120例子宫内膜癌患者中,发生淋巴结转移28例;淋巴结转移组血清载脂蛋白AI低于非淋巴结转移组,鳞状细胞癌抗原表达水平高于非淋巴结转移组（P＜0.05）;经ROC曲线分析,血清载脂蛋白AI联合鳞状细胞癌抗原预测子宫内膜癌患者发生淋巴结转移的AUC为0.910。结论：血清载脂蛋白AI、鳞状细胞癌抗原与子宫内膜癌的不同临床分期、病理分级、子宫肌层浸润程度有关,术前检测两者表达水平有助于淋巴结转移的预测,对于指导临床治疗具有积极作用。     

A simplified and efficient procedure for the purification of apolipoprotein AI from human serum high-density lipoprotein-3 by preparative isoelectric focussing on polyacrylamide gel beads

An improved method is described for the purification of milligram amounts of apolipoprotein AI from serum apo-HDL₃ by isoelectric focussing on polyacrylamide gel beads. The procedure involves a single focussing over a narrow (1.3 unit) pH gradient, and permits isolation of apo-AI of exceptional purity and in high yield (75% recovery of HDL₃ protein, ca. 50% corresponding to pure apo-AI). The electrophoretic mobility, pI values, molecular weight, antigenicity and amino acid composition of such apo-AI were indistinguishable from those reported in the literature. A rabbit antiserum to apo-AI isolated by focusing exhibited similar immunological reactivity to one prepared from an antigen isolated by gel filtration chromatography; moreover, apo-AI purified by the respective procedures reacted identically with both antisera. We conclude that isoelectric focussing on a support of polyacrylamide gel beads (as Bio-Gel P60) presents certain advantages for the isolation of highly purified apo-AI over both conventional chromatographic procedures and isoelectric focussing on a Sephadex support.     

Laboratory and field evaluation of insect repellents as oviposition deterrents against the mosquito Aedes albopictus

Three experimental approaches were used to evaluate the oviposition deterrency of three insect repellents, AI3-35765, AI3-37220 (piperidine compounds), and the standard N,N-diethyl-3-methylbenzamide (deet) to the mosquito Aedes albopictus Skuse (Diptera: Culicidae). Against laboratory-reared Ae. albopictus gravid females, the EC50 values of AI3-37220, AI3-35765 and deet were 0.004%, 0.008% and 0.011% in laboratory cages and 0.004%, 0.01% and 0.009% in an outdoor screened cage. For a natural population of Ae. albopictus tested in the field, the EC50 values were determined as 0.004%, 0.008% and 0.001%, respectively. Ageing concentrations of 0.1% of each repellent provided >50% effective oviposition deterrency against the laboratory population of Ae. albopictus for 13 days in laboratory cages, for 15 days in the outdoor cage, and for 21 days against field population of Ae. albopictus in Florida. These topical skin repellents are effective oviposition deterrents for Ae. albopictus when employed at relatively low application rates.     

Adenine recognition: a motif present in ATP-, CoA-, NAD-, NADP-, and FAD-dependent proteins.

Adenosine triphosphate (ATP) plays an essential role in energy transfer within the cell. In the form of NAD, adenine participates in multiple redox reactions. Phosphorylation and ATP-hydrolysis reactions have key roles in signal transduction and regulation of many proteins, especially enzymes. In each cell, proteins with many different functions use adenine and its derivatives as ligands; adenine, of course, is present in DNA and RNA. We show that an adenine binding motif, which differs according to the backbone chain direction of a loop that binds adenine (and in one variant by the participation of an aspartate side-chain), is common to many proteins; it was found from an analysis of all adenylate-containing protein structures from the Protein Data Bank. Indeed, 224 protein-ligand complexes (86 different proteins) from a total of 645 protein structure files bind ATP, CoA, NAD, NADP, FAD, or other adenine-containing ligands, and use the same structural elements to recognize adenine, regardless of whether the ligand is a coenzyme, cofactor, substrate, or an allosteric effector. The common adenine-binding motif shown in this study is simple to construct. It uses only (1) backbone polar interactions that are not dependent on the protein sequence or particular properties of amino acid side-chains, and (2) nonspecific hydrophobic interactions. This is probably why so many different proteins with different functions use this motif to bind an adenylate-containing ligand. The adenylate-binding motif reported is present in "ancient proteins" common to all living organisms, suggesting that adenine-containing ligands and the common motif for binding them were exploited very early in evolution. The geometry of adenine binding by this motif mimics almost exactly the geometry of adenine base-pairing seen in DNA and RNA.     

胆固醇逆转运相关蛋白基因在骨骼肌的表达

构建载脂蛋白ＡＩ（ａｐｏＡＩ）、载脂蛋白Ｅ（ａｐｏＥ）和卵磷脂胆固醇酰基转移酶（ＬＣＡＴ）的病毒或非病毒载体,分别转染离体成肌细胞或直接注入小鼠骨骼肌,观察其表达程序及分泌人血等情况,探讨借用骨骼肌异源表达这些在胆固醇逆转运中起关键作用的重要候选基因,进而发展一种简易安全基因治疗方法的可能性。结果显示,质粒表达载体ｐＣＭＶａｐｏＥ３和重组腺病毒载体Ａｄ－ＲＳＶ－ａｐｏＡＩ在原代培养小鼠成肌细胞和成     

Approval policies for modifications to machine learning‐based software as a medical device: A study of bio‐creep

Successful deployment of machine learning algorithms in healthcare requires careful assessments of their performance and safety. To date, the FDA approves locked algorithms prior to marketing and requires future updates to undergo separate premarket reviews. However, this negates a key feature of machine learning—the ability to learn from a growing dataset and improve over time. This paper frames the design of an approval policy, which we refer to as an automatic algorithmic change protocol (aACP), as an online hypothesis testing problem. As this process has obvious analogy with noninferiority testing of new drugs, we investigate how repeated testing and adoption of modifications might lead to gradual deterioration in prediction accuracy, also known as “biocreep” in the drug development literature. We consider simple policies that one might consider but do not necessarily offer any error‐rate guarantees, as well as policies that do provide error‐rate control. For the latter, we define two online error‐rates appropriate for this context: bad approval count (BAC) and bad approval and benchmark ratios (BABR). We control these rates in the simple setting of a constant population and data source using policies aACP‐BAC and aACP‐BABR, which combine alpha‐investing, group‐sequential, and gate‐keeping methods. In simulation studies, bio‐creep regularly occurred when using policies with no error‐rate guarantees, whereas aACP‐BAC and aACP‐BABR controlled the rate of bio‐creep without substantially impacting our ability to approve beneficial modifications.     

Evidence of expression variation and allelic imbalance in Crohn's disease susceptibility genes NOD2 and ATG16L1 in human dendritic cells

Human dendritic cells (DCs) play an important role in induction and progression of Crohn's disease (CD). Accumulating evidence suggests that viral infection is required to trigger CD pathogenesis in genetically predisposed individuals. NOD2 and ATG16L1 are among the major CD susceptibility genes implicated in impaired immune response to bacterial infection. In this study, we investigated gene expression and allelic imbalance (AI) of NOD2 and ATG16L1 using common variants in human monocyte-derived DCs. Significant AI was observed in ~ 40% and ~ 70% of NOD2 and ATG16L1 heterozygotes, respectively (p < 0.05). AI of NOD2 was inversely associated with its expression level (p = 0.015). No correlation was detected between gene expression and AI for ATG16L1. When infected with Newcastle Disease Virus (NDV), NOD2 expression in DCs was induced about four-fold (p < 0.001), whereas ATG16L1 expression was not affected (p = 0.88). In addition, NDV infection tended to lower the variance in AI among DC populations for the NOD2 gene (p = 0.05), but not the ATG16L1 gene (p = 0.32). Findings of a simulation study, aimed to verify whether the observed variation in gene expression and AI is a result of sample-to-sample variability or experimental measurement error, suggested that NOD2 AI is likely to result from a deterministic event at a single cell level. Overall, our results present initial evidence that AI of the NOD2 and ATG16L1 genes exists in populations of human DCs. In addition, our findings suggest that viral infection may regulate NOD2 expression.