BACKGROUND & AIMS: :Auto-anti-idiotypic mechanisms can regulate the protective immune response against Schistosoma mansoni. Anti-idiotypic responses were stimulated by immunization of mice either with nonspecifically induced lymphoblasts, produced with Con A, or with Ag-induced lymphoblasts bearing specific idiotypic receptors. The effect of the induced anti-idiotypic response upon clonotypic cellular reactivity was assessed in vitro through the suppression of antigen-mediated blast transformation by cloned T cells and in vivo by suppression of resistance to S. mansoni and delayed-type hypersensitivity responses against specific Ag. Differential regulation of humoral immune responses was studied at the levels of specific epitopic recognition, the expression of specific Id, and the production of anti-idiotypic responses directed against mAb bearing specific Id. Anti-idiotypic sensitization resulted in variable (10 to 90%) suppression of the immune response to discrete antigenic epitopes, the expression of specific idiotypic phenotypes, and anti-idiotypic, antiparatopic responses against T cell clonotypes and antibody idiotypic phenotypes. In vitro admixture and in vivo challenge studies resulted in consonant differential suppression. Thus idiotypic regulation can mold the fine specificities of the protective immune response to S. mansoni at the clonal level and may provide an approach to optimize the expression and assessment of resistance.

背景与目标： 自动抗独特型机制可以调节针对曼氏血吸虫的保护性免疫反应。通过用Con A产生的非特异性诱导的淋巴母细胞，或用Ag诱导的携带特定独特型受体的淋巴母细胞免疫小鼠，可以刺激抗独特型应答。在体外通过抑制克隆的T细胞对抗原介导的胚泡转化的抑制来评估诱导的抗独特型反应对克隆型细胞反应性的影响，并在体内通过抑制对曼氏沙门氏菌的抗性和针对特定Ag的迟发型超敏反应来评估体内诱导的抗独特型反应的作用。 。研究了在特定表位识别，特定Id的表达以及针对带有特定Id的mAb的抗独特型反应水平上的体液免疫应答的差异调节。抗独特型致敏作用导致对离散抗原表位的免疫反应，特异性独特型表型的表达以及针对T细胞克隆型和抗体独特型表型的抗独特型，抗副反应的变化受到抑制（10％至90％）。体外混合和体内攻击研究导致辅音差异抑制。因此，独特型调节可以在克隆水平上塑造对曼氏链球菌的保护性免疫应答的优良特异性，并且可以提供优化抗性表达和评估的方法。

BACKGROUND & AIMS: :Serum samples from 224 aboriginal Amazonian Indians were tested for antibodies to HTLV-III/LAV by an indirect immunofluorescence (IF) assay. 9 individuals (4%), 5 of them female, were seropositive by IF and by confirmatory western blotting and radioimmunoprecipitation tests. 3 of the positive sera were collected in 1968. HTLV-III/LAV seropositivity rates varied among the ethnic groups and ranged from 13.3% among the Pemon Indians to 3.3% among the Yanoama tribe. The titres of HTLV-III/LAV antibodies ranged from 1/40 to 1/320. All individuals tested were apparently healthy at the time of the study. None of 211 randomly chosen, healthy blood donors from Venezuelan cities had antibodies to HTLV-III/LAV. The prevalence of specific antibodies among Amazonian Indians suggests the HTLV-III/LAV or a closely related cross-reactive virus may be endemic in this area. The findings also indicate that this virus is indigenous in non-negroid Latin American and negroid tropical populations.

背景与目标： ：通过间接免疫荧光（IF）分析测试了来自224名原住民亚马逊印第安人的血清样品中HTLV-III / LAV的抗体。通过IF，证实性Western印迹和放射免疫沉淀试验，有9名个体（4％），其中5名是女性血清阳性。 1968年收集了3份阳性血清。HTLV-III/ LAV血清阳性率在不同种族之间有所不同，范围从Pemon印第安人的13.3％到Yanoama部落的3.3％。 HTLV-III / LAV抗体的滴度范围为1/40至1/320。在研究时，所有接受测试的人显然都是健康的。委内瑞拉城市的211名随机抽取的健康献血者中没有一个具有针对HTLV-III / LAV的抗体。亚马逊印第安人中特异性抗体的流行表明，HTLV-III / LAV或密切相关的交叉反应性病毒可能是该地区的地方病。研究结果还表明，该病毒是非黑人拉丁美洲和黑人热带人群中的土著病毒。

BACKGROUND & AIMS: :Aldosterone is synthesized in the zona glomerulosa of the adrenal cortex under primary regulation by the renin-angiotensin system. Angiotensin II (A-II) acts through the angiotensin types 1 and 2 receptors (AT1R and AT2R). A-II is metabolized in different tissues by various enzymes to generate two heptapeptides A-III and angiotensin 1-7, which can then be catabolized into smaller peptides. A-II was more potent than A-III in stimulating aldosterone secretion in the adrenocortical cell line HAC15, and A-II, but not A-III, stimulated cortisol secretion. A-II stimulated mRNA expression of steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase, CYP11B1, and CYP11B2, whereas A-III stimulated 3β-hydroxysteroid dehydrogenase, CYP11B1, and CYP11B2 but decreased the expression of CYP17A1 required for cortisol synthesis. The stimulation of aldosterone secretion by A-II and A-III was blocked by the AT1R receptor blocker, losartan, but not by an AT2R blocker. A-II was rapidly metabolized by the HAC15 cells to mainly to angiotensin 1-7, but not to A-III, and disappeared from the supernatant within 6 h. A-III was metabolized rapidly and disappeared within 1 h. In conclusion, A-II was not converted to A-III in the HAC15 cell and is the more potent stimulator of aldosterone secretion and cortisol of the two. A-III stimulated aldosterone secretion but not cortisol secretion.

背景与目标： ：醛固酮是在肾素-血管紧张素系统的主要调控下在肾上腺皮质的肾小球中合成的。血管紧张素II（A-II）通过1型和2型血管紧张素受体（AT1R和AT2R）起作用。 A-II通过各种酶在不同的组织中代谢，生成两种七肽A-III和血管紧张素1-7，然后可以将其分解为较小的肽。在刺激肾上腺皮质细胞系HAC15中的醛固酮分泌方面，A-II比A-III更有效，而A-II（而非A-III）刺激皮质醇分泌。 A-II刺激类固醇生成的急性调节蛋白，3β-羟基类固醇脱氢酶，CYP11B1和CYP11B2的mRNA表达，而A-III刺激3β-羟基类固醇脱氢酶，CYP11B1和CYP11B2但降低了皮质醇合成所需的CYP17A1的表达。 A-II和A-III对醛固酮分泌的刺激被AT1R受体阻滞剂氯沙坦阻断，但未被AT2R阻滞剂阻断。 A-II被HAC15细胞快速代谢，主要代谢为血管紧张素1-7，但不代谢为A-III，并在6小时内从上清液中消失。 A-III快速代谢并在1​​小时内消失。总之，A-II在HAC15细胞中并未转化为A-III，而是对两者的醛固酮分泌和皮质醇更有效的刺激剂。 A-III刺激醛固酮分泌，但不刺激皮质醇分泌。

BACKGROUND & AIMS: :gamma-Secretase is a protease complex, which catalyzes the final of two subsequent cleavages of the beta-amyloid precursor protein (APP) to release the amyloid-beta peptide (Abeta) implicated in Alzheimer's disease (AD) pathogenesis. In human cells, six gamma-secretase complexes exist, which are composed of either presenilin (PS) 1 or 2, the catalytic subunit, nicastrin, PEN-2, and either APH-1a (as S or L splice variants) or its homolog APH-1b. It is not known whether and how different APH-1 species contribute to the pathogenic activity of gamma-secretase complexes with familial AD (FAD)-associated mutant PS. Here we show that all known gamma-secretase complexes are active in APP processing and that all combinations of APH-1 variants with either FAD mutant PS1 or PS2 support pathogenic Abeta(42) production. Since our data suggest that pathogenic gamma-secretase activity cannot be attributed to a discrete gamma-secretase complex, we propose that all gamma-secretase complexes have to be explored and evaluated for their potential as AD drug target.

背景与目标： ：-Secretase是一种蛋白酶复合物，它催化β-淀粉样前体蛋白（APP）的两个后续切割的最终过程，以释放与阿尔茨海默氏病（AD）发病机理有关的淀粉样β-肽（Abeta）。在人类细胞中，存在六种γ-分泌酶复合物，它们由早老素（PS）1或2，催化亚基，尼卡斯特林，PEN-2和APH-1a（作为S或L剪接变体）或其同系物组成APH-1b。尚不知道不同的APH-1物种是否以及如何与家族性AD（FAD）相关突变PS共同促进γ-分泌酶复合物的致病活性。在这里，我们显示所有已知的伽玛分泌酶复合物在APP处理中均具有活性，并且APH-1变体与FAD突变PS1或PS2的所有组合均支持病原性Abeta（42）生产。由于我们的数据表明致病性γ-分泌酶活性不能归因于离散的γ-分泌酶复合物，因此我们建议必须探索和评估所有γ-分泌酶复合物作为AD药物靶标的潜力。

BACKGROUND & AIMS: :The synthesis and characterisation of iridium(III) bis(2-(2,4-difluorophenyl)pyridinato-N, C2')-2(4-carboxylphenyl)imidazo[4,5-f][1,10]phenanthroline perchlorate, [Ir(dfpp)(2)(picCOOH)](+) and its octaarginine conjugate [Ir(dfpp)(2)(picCONH-Arg(8))](9+) are reported. Both complex and conjugate exhibit intense and long-lived luminescence, which is O(2) and pH sensitive. Conjugation to the polyarginine peptide renders the complex very water soluble. The uptake of the parent iridium(III) complex and conjugate are compared in two mammalian cell lines; SP2 myeloma and Chinese hamster ovary (CHO). Both complexes internalise into the cytoplasm, however dye uptake rate and distribution vary with peptide conjugation and with cell identity. Whereas transmembrane transport is thought to have been facilitated by the dimethyl sulfoxide (DMSO) used as co-solvent (0.05% v/v) for the parent complex, the octaarginine, the dye-conjugate (iridium-R(8)) is membrane permeable in water only. Both complexes exhibit high cytotoxicity, evident through blebbing and vacuole formation within living cells, indicative of apoptosis, within 30min of exposure to the probe. The IC(50) recorded for the cells in the dark was independent, in the case of the parent complex, of the identity of the cell, with IC(50) of 84.8μM and 88μM respectively for SP2 and CHO cells. The IC(50) approximately doubled for the polyarginine conjugate and displayed a significant dependence on cell type with IC(50) of 35μM and 54.1μM respectively for SP2 and CHO cells. These IC(50) values were recorded in the dark. However under irradiation cell death is considerably faster. Evidence from imaging suggests that the conjugate penetrates the nucleus whereas the parent does not, indicating that nuclear penetration may play a role in cytotoxicity.

背景与目标： ：铱（III）双（2-（2,4-二氟苯基）吡啶基-N，C2'）-2（4-羧基苯基）咪唑并[4,5-f] [1,10]菲咯啉高氯酸盐的合成与表征，[Ir（dfpp）（2）（picCOOH）]（）及其八精氨酸共轭物[Ir（dfpp）（2）（picCONH-Arg（8））]（9）被报道。复杂和共轭物都显示强烈和长寿命的发光，这是O（2）和pH敏感的。与聚精氨酸肽的缀合使得该复合物非常水溶性。在两种哺乳动物细胞系中比较了母体铱（III）配合物和结合物的摄取。 SP2骨髓瘤和中国仓鼠卵巢（CHO）。两种复合物都内化到细胞质中，但是染料吸收率和分布随肽结合和细胞身份而变化。认为作为母体配合物的助溶剂（0.05％v / v）的二甲基亚砜（DMSO）促进了跨膜运输，而八精氨酸，染料-缀合物（铱-R（8））是膜仅在水中可渗透。两种复合物均表现出高的细胞毒性，这在暴露于探针后30分钟内在活细胞内起泡和形成液泡很明显，表明细胞凋亡。在亲本复合物的情况下，黑暗中细胞的IC（50）与细胞的身份无关，SP2和CHO细胞的IC（50）分别为84.8μM和88μM。对于聚精氨酸缀合物，IC（50）大约增加了一倍，并显示出对细胞类型的显着依赖性，SP2和CHO细胞的IC（50）分别为35μM和54.1μM。这些IC（50）值在黑暗中记录。然而，在辐射下，细胞死亡要快得多。影像学证据表明，结合物穿透核，而亲本没有穿透核，表明核穿透可能在细胞毒性中起作用。

BACKGROUND & AIMS: BACKGROUND:Cardiovascular patients are likely to have an impaired health-related quality of life (HRQoL) due to functional and psycho-social limitations. The main objective of this study was to assess the distribution of HRQoL scores in coronary heart disease (CHD) patients across 22 European countries and to identify factors associated with the variation between patients. METHODS:Data from the EUROASPIRE III survey (European Action on Secondary and Primary Prevention by Intervention to Reduce Events), on 8734 patients, were used. Patients with a diagnosis of CHD (coronary artery bypass graft (CABG), percutaneous coronary intervention (PCI), acute myocardial infarction (AMI) or myocardial ischemia) were interviewed and examined at least 6 months after their acute coronary event. Quality of life of each patient was measured using 2 standardized questionnaires: the EuroQoL-5D (EQ-5D) and the 12-item short-form health survey (SF-12v2). RESULTS:HRQoL values differed significantly across countries. Lower HRQoL estimates were found in women, older patients, less educated patients, patients with myocardial infarction or ischemia as recruiting diagnosis, patients with a history of stroke and patients who suffered from a recurring CHD event. In addition, HRQoL was significantly associated with current smoking, central obesity, lack of exercise and inappropriate HbA1c control in patients with diabetes. Furthermore the number of risk factors is inversely associated with HRQoL. CONCLUSION:Overall, a large heterogeneity was observed in HRQoL values between countries and patient groups. There seems to be a significant association between quality of life and patient characteristics with lifestyle risk factors as important determinants of HRQoL.

背景与目标： 背景：由于功能和心理社会方面的限制，心血管疾病患者的健康相关生活质量（HRQoL）可能受损。这项研究的主要目的是评估HRQoL分数在22个欧洲国家中的冠心病（CHD）患者中的分布，并确定与患者之间差异相关的因素。
方法：使用EUROASPIRE III调查（欧洲通过干预减少事件的二级和一级预防行动）对8734例患者的数据。诊断为冠心病（冠状动脉搭桥术（CABG），经皮冠状动脉介入治疗（PCI），急性心肌梗塞（AMI）或心肌缺血）的患者在其急性冠状动脉事件发生后至少6个月进行了访谈和检查。每个患者的生活质量使用2份标准问卷进行测量：EuroQoL-5D（EQ-5D）和12个项目的简短健康调查（SF-12v2）。
结果：HRQoL值在不同国家之间存在显着差异。在女性，年龄较大的患者，受教育程度较低的患者，患有心肌梗塞或局部缺血的患者（作为招募诊断），有中风病史的患者以及患有复发性冠心病事件的患者中发现较低的HRQoL估计值。此外，HRQoL与糖尿病患者当前的吸烟，中心性肥胖，缺乏运动和不适当的HbA1c控制密切相关。此外，危险因素的数量与HRQoL成反比。
结论：总体而言，国家和患者群体之间的HRQoL值存在很大的异质性。生活质量和患者特征与作为HRQoL的重要决定因素的生活方式危险因素之间似乎存在显着联系。

BACKGROUND & AIMS: :We have studied by electron microscopy a fascinating series of antidansyl monoclonal antibodies developed by Dangl et al. (Cytometry 2, 395-401, 1982) which have the same variable domain but different constant domains. Three of the four subclasses of mouse IgG were represented, IgG1, IgG2a and IgG2b. Previously, Oi et al. (Nature 307, 136-140, 1984) had examined the flexibilities of these antibodies by time-resolved fluorescence depolarization and found that IgG1 was least flexible, IgG2a was intermediate and IgG2b was the most flexible. In this communication we examine the conformations of circular complexes formed between these antibodies and a bivalent hapten, bis-dansyl cadaverine. The circular complexes were predominantly composed of two antibodies linked into a ring by two bivalent haptens, and are referred to as dimers, since only the antibody molecules are seen with the electron microscope. A few trimers and an occasional tetramer were also present in these preparations. For the least flexible IgG1, almost all (greater than 99%) of the circular dimers were "open-hinge" complexes with a hinge angle between the Fab arms of 100-120 degrees. For the intermediate IgG2a, most of the dimers were "open-hinge" complexes, but a larger percentage, 4 to 5%, had closed hinges with a hinge angle approaching 0 degrees. For the most flexible IgG2b, over 40% of the dimers were "closed-hinge" complexes. A model is proposed to explain these differences based upon orientation of the hapten in the combining site and differences in hinge structure.

背景与目标： ：我们已经通过电子显微镜研究了由Dangl等人开发的一系列令人着迷的抗丹磺酰单克隆抗体。 （Cytometry 2，395-401，1982），它们具有相同的可变域，但具有不同的恒定域。代表了小鼠IgG的四个亚类中的三个，即IgG1，IgG2a和IgG2b。以前，Oi等人。 （Nature 307，136-140，1984）通过时间分辨荧光去极化检查了这些抗体的柔韧性，发现IgG1的柔韧性最低，IgG2a的柔韧性中等，而IgG2b的柔韧性最高。在本交流中，我们研究了这些抗体与二价半抗原，双丹磺酰尸胺之间形成的环状复合物的构象。环状复合物主要由通过两个二价半抗原连接成环的两种抗体组成，被称为二聚体，因为在电子显微镜下只能看到抗体分子。在这些制剂中还存在一些三聚体和偶尔的四聚体。对于最不灵活的IgG1，几乎所有（大于99％）的圆形二聚体都是“开放铰链”复合物，Fab臂之间的铰链角为100-120度。对于中间体IgG2a，大多数二聚体是“开放铰链”复合物，但较大的百分比（4％至5％）具有闭合的铰链，铰链角接近0度。对于最灵活的IgG2b，超过40％的二聚体是“封闭铰链”复合物。提出了一个模型来解释这些差异，这些差异是基于半抗原在结合位点的方向和铰链结构的差异而得出的。

BACKGROUND & AIMS: :The replication of influenza virus RNA in vitro has been studied by cell fractionation of MDCK-infected cells and characterization of in vitro synthesized RNA. Analysis of the RNA product polarity by liquid hybridization to excess single-stranded DNA probes shows that only the RNP complexes present in the nuclear matrix fraction are able to synthesize negative-polarity RNA. This RNA product has been characterized as authentic vRNA by size analysis, RNase-protection by unlabelled, positive-polarity riboprobes and T1-fingerprinting. Priming the in vitro reaction with ApG stimulates preferentially the synthesis of positive-polarity RNA, while ApGpU stimulates both positive and negative-polarity RNA synthesis.

背景与目标： ：已通过MDCK感染细胞的细胞分级分离和体外合成RNA的表征研究了流感病毒RNA的体外复制。通过与过量的单链DNA探针进行液体杂交来分析RNA产物的极性表明，只有存在于核基质组分中的RNP复合物才能合成负极性RNA。通过大小分析，未标记的正极性核糖核糖核酸和T1指纹图谱已将该RNA产物表征为真正的vRNA。用ApG引发体外反应优先刺激正极性RNA的合成，而ApGpU刺激正极性和负极性RNA的合成。

BACKGROUND & AIMS: :We sporadically experienced three paediatric cases of atypical upswing at the initial part of phase III in the capnograms via side-stream capnometer immediately following endotracheal intubation and mechanical ventilation. No fault was found in the monitor or anaesthetic system including breathing circuits, carbon dioxide (CO2 ) sampling tube, water trap, and unidirectional valves. The upsurge of CO2 disappeared with increasing the respiratory rate; however, it reappeared with decreasing the respiratory rate and vice versa. We experimentally reproduced the phenomenon of overshooting CO2 measurement after the luer lock connection of the sample gas line at the water trap had been unscrewed a little bit.

背景与目标： ：我们在气管插管和机械通气后立即通过侧流式二氧化碳监测仪散发了3例小儿在非典型性上升阶段的二氧化碳监测图的III期初期的案例。在监测器或麻醉系统中未发现故障，包括呼吸回路，二氧化碳（CO2）采样管，集水器和单向阀。随着呼吸频率的增加，二氧化碳的升高消失了。然而，它随着呼吸频率的降低而重新出现，反之亦然。我们实验性地重现了旋开集水器处的样气管线的鲁尔锁连接后，CO2测量值超调的现象。

BACKGROUND & AIMS: :We introduce a new economic, convenient and general assay principle based on the reversible interaction of water-soluble macrocycles and fluorescent dyes. We show that amino acid decarboxylase activity can be continuously monitored by measuring changes in fluorescence, which result from the competition of the enzymatic product and the dye for forming a complex with a cucurbituril or calixarene macrocycle. The new assay provides a complementary method to the use of antibodies, radioactive markers and labeled substrates.

背景与目标： ：我们基于水溶性大环化合物和荧光染料的可逆相互作用，引入了一种新的经济，方便，通用的测定原理。我们表明，可以通过测量荧光的变化来连续监测氨基酸脱羧酶的活性，荧光的变化是由于酶产物与染料竞争而形成的，其与葫芦科草或杯芳烃大环形成了复合物。新的检测方法为抗体，放射性标记物和标记底物的使用提供了一种补充方法。

BACKGROUND & AIMS: :The versatile combination of affinity purification and mass spectrometry (AP-MS) has recently been applied to the detailed characterization of many protein complexes and large protein-interaction networks. The combination of AP-MS with other techniques, such as biochemical fractionation, intact mass measurement and chemical crosslinking, can help to decipher the supramolecular organization of protein complexes. AP-MS can also be combined with quantitative proteomics approaches to better understand the dynamics of protein-complex assembly.

背景与目标： ：亲和纯化和质谱分析（AP-MS）的多功能组合最近已用于许多蛋白质复合物和大型蛋白质相互作用网络的详细表征。 AP-MS与其他技术（例如生化分级分离，完整质量测量和化学交联）的结合可以帮助破译蛋白质复合物的超分子组织。 AP-MS也可以与定量蛋白质组学方法结合使用，以更好地了解蛋白质复合物组装的动力学。

BACKGROUND & AIMS: :Mononuclear complexes [Cu(Itpy)X(H2O)]X (Itpy--imidazole terpyridine, X--NO3 1 and X--ClO4 2) have been synthesized and characterized. Single crystal X-ray diffraction of complex 1 shows distorted octahedral geometry around the copper (II) ion. Presence of multiple hydrogen bonding network in the molecule results in anti-parallel stacking of the molecule. Both the complexes show dual mode of binding to DNA. Both the complexes have been found to bring about DNA cleavage in the presence of H2O2 and show potent cytotoxicity towards lung carcinoma cell line. The ability of the two complexes to induce apoptosis has been investigated by using combination of nuclear stains. FACS analysis shows that both the complexes bring about cell cycle arrest at 2.5 μM concentration.

背景与目标： ：已经合成并表征了单核络合物[Cu（Itpy）X（H2O）] X（Itpy-咪唑联吡啶，X-NO3 1和X-ClO4 2）。配合物1的单晶X射线衍射显示铜（II）离子周围的八面体几何形状失真。分子中存在多个氢键网络会导致分子反平行堆积。两种复合物均显示出与DNA结合的双重模式。已经发现两种复合物在H 2 O 2存在下引起DNA切割，并且显示出对肺癌细胞系的有效细胞毒性。通过使用核染色剂的组合已经研究了两种复合物诱导凋亡的能力。 FACS分析表明，两种复合物均以2.5μM的浓度引起细胞周期停滞。

BACKGROUND & AIMS: :The type III secretion system (T3SS) of gram-negative bacteria involves dedicated protein translocation machinery that directly injects proteins into target cells. Pathogenic bacteria already benefit from this unique system. The successful functional cloning of this useful tool into non-pathogenic bacteria would help establish novel clinical and basic biotechnology strategies in areas such as vaccine administration, the development of screening systems for anti-T3SS drugs and the target-specific delivery of bioactive compounds. In this study, we successfully cloned the Vibrio parahaemolyticus T3SS1 genetic locus into a non-pathogenic Escherichia coli K-12 strain. Assays performed here revealed that the T3SS1 cloned into the E. coli K-12 strain has the ability to translocate V. parahaemolyticus T3SS1 secreted proteins. Importantly, we also observed this system to allow the E. coli K-12 strain to inject foreign protein, as well as the V. parahaemolyticus T3SS effector, into cultured cells. These results demonstrate a prospective useful tool with experimental and therapeutic applications.

背景与目标： ：革兰氏阴性细菌的III型分泌系统（T3SS）涉及专用的蛋白质转运机制，可直接将蛋白质注射到靶细胞中。致病细菌已经从这种独特的系统中受益。将该有用工具成功地功能克隆到非致病性细菌中，将有助于在疫苗管理，抗T3SS药物筛选系统的开发以及生物活性化合物的靶标特异性递送等领域建立新颖的临床和基础生物技术策略。在这项研究中，我们成功地将副溶血性弧菌T3SS1遗传基因座克隆到了非致病性大肠杆菌K-12菌株中。此处进行的分析表明，克隆到大肠杆菌K-12菌株中的T3SS1具有转移副溶血弧菌T3SS1分泌蛋白的能力。重要的是，我们还观察到该系统允许大肠杆菌K-12菌株向培养的细胞中注入外源蛋白以及副溶血弧菌T3SS效应子。这些结果证明了具有实验和治疗应用前景的有用工具。

BACKGROUND & AIMS: :A bacterioferritin was recently isolated from the anaerobic sulphate-reducing bacterium Desulfivibrio desulfuricans ATCC 27774 [Romão et al. (2000) Biochemistry 39, 6841-6849]. Although its properties are in general similar to those of the other bacterioferritins, it contains a haem quite distinct from the haem B, found in bacterioferritins from aerobic organisms. Using visible and NMR spectroscopies, as well as mass spectrometry analysis, the haem is now unambiguously identified as iron-coproporphyrin III, the first example of such a prosthetic group in a biological system. This unexpected finding is discussed in the framework of haem biosynthetic pathways in anaerobes and particularly in sulphate-reducing bacteria.

背景与目标： ：最近从减少厌氧硫酸盐的细菌Desulfivibrio desulfuricans ATCC 27774中分离出一种细菌铁蛋白。 （2000）Biochemistry 39，6841-6849]。尽管其性质通常与其他细菌铁蛋白相似，但它所包含的血红素与好氧生物的细菌铁蛋白中发现的血红素B完全不同。使用可见光谱和NMR光谱以及质谱分析，血红素现在被明确鉴定为铁-卟啉III，这是生物系统中此类辅基的第一个实例。在厌氧菌，尤其是硫酸盐还原菌中的血红素生物合成途径的框架中讨论了这一出乎意料的发现。

BACKGROUND & AIMS: :The Escherichia coli DNA polymerase III tau and gamma subunits are single-strand DNA-dependent ATPases (the latter requires the delta and delta' subunits for significant ATPase activity) involved in loading processivity clamp beta. They are homologous to clamp-loading proteins of many organisms from phages to humans. Alignment of 27 prokaryotic tau/gamma homologs and 1 eukaryotic tau/gamma homolog has refined the sequences of nine previously defined identity and functional motifs. Mutational analysis has defined highly conserved residues required for activity in vivo and in vitro. Specifically, mutations introduced into highly conserved residues within three of those motifs, the P loop, the DExx region, and the SRC region, inactivated complementing activity in vivo and clamp loading in vitro and reduced ATPase catalytic efficiency in vitro. Mutation of a highly conserved residue within a fourth motif, VIc, inactivated clamp-loading activity and reduced ATPase activity in vitro, but the mutant gene, on a multicopy plasmid, retained complementing activity in vivo and the mutant gene also supported apparently normal replication and growth as a haploid, chromosomal allele.

背景与目标： ：大肠杆菌DNA聚合酶III tau和gamma亚基是单链DNA依赖的ATPase（后者需要具有明显的ATPase活性的delta和delta'亚基），参与上样性钳制β的过程。它们与将多种生物的蛋白质从噬菌体夹入人类的过程同源。 27个原核tau /γ同源物和1个真核tau /γ同源物的比对已完善了9个先前定义的同一性和功能性基序的序列。突变分析已定义了体内和体外活性所需的高度保守的残基。具体而言，将突变引入到其中三个基序（P环，DExx区和SRC区）中高度保守的残基中，使体内的补体活性失活，并在体外钳夹负载，并降低体外的ATPase催化效率。在体外，第四个基序VIc中高度保守的残基突变，失活了钳位活性并降低了ATPase活性，但在多拷贝质粒上的突变基因在体内保留了互补活性，并且该突变基因还明显支持正常复制和生长为单倍体，染色体等位基因。