BACKGROUND & AIMS: :Inverted duplicates are a type of repetitive DNA motifs consist of two copies of reverse complementary sequences separated by a spacer sequence. They can lead to genome instability and many may have no function, but some functional small RNAs are processed from hairpins transcribed from these elements. It is not clear whether the pervasive numbers of such elements in genomes, especially those of mammals, is the result of high generation rates of neutral or slightly deleterious duplication events or positive selection for functionality. To test the functionality of intergenic inverted duplicates without known functions, we used mirror duplicates, a type of repetitive DNA motifs with few reported functions and little potential to form hairpins when transcribed, as a nonfunctional control. We identified large numbers of inverted duplicates within intergenic regions of human and mouse genomes, as well as 19 other vertebrate genomes. Structure characterization of these inverted duplicates revealed higher proportion to form stable hairpins compared with converted mirror duplicates, suggesting that inverted duplicates may produce hairpin RNAs. Expression profiling across tissues demonstrated that 7.8% of human and 5.7% of mouse inverted duplicates were expressed even under strict criteria. We found that expressed inverted duplicates were more likely to be structurally stable than both unexpressed inverted duplicates and expressed converted mirror duplicates. By dating inverted duplicates in the vertebrate phylogenetic tree, we observed higher conservation of inverted duplicates than mirror duplicates. These observations support the notion that expressed inverted duplicates may be functional through forming hairpin RNAs.

背景与目标： : 反向重复序列是一种重复的DNA基序，由两个由间隔区序列分隔的反向互补序列的拷贝组成。它们可能导致基因组不稳定，许多可能没有功能，但是一些功能性小rna是从这些元件转录的发夹中加工出来的。尚不清楚基因组中此类元素的普遍数量，尤其是哺乳动物的基因组中，是否是中性或轻微有害复制事件的高产生率或功能的积极选择的结果。为了测试没有已知功能的基因间倒置重复项的功能，我们使用了镜像重复项 (一种重复DNA基序，其报道功能很少，转录时形成发夹的潜力很小) 作为非功能对照。我们在人和小鼠基因组的基因间区域以及其他19个脊椎动物基因组中鉴定出大量的反向重复。与转换的镜像副本相比，这些倒置副本的结构表征显示出形成稳定发夹的比例更高，这表明倒置副本可能产生发夹rna。跨组织的表达谱表明，即使在严格的标准下，也表达了人类的7.8% 和小鼠倒置副本的5.7%。我们发现，表达的倒置重复项比未表达的倒置重复项和表达的转换镜像重复项更可能在结构上稳定。通过对脊椎动物系统发育树中的倒置重复物进行定年，我们观察到倒置重复物的保守性高于镜像重复物。这些观察结果支持以下观点: 表达的反向重复可能通过形成发夹rna而起作用。

BACKGROUND & AIMS: :Two strains of Aeromonas salmonicida, YK and BG, were isolated from largemouth bronze gudgeon and northern whitefish in China, and identified as A. salmonicida subsp. salmonicida based on phylogenetic analysis of vapA and 16S rRNA gene sequences. YK and BG originated from freshwater fish, one of which belonged to the cyprinid family, and the strains showed a difference in virulence. Subsequently, we performed whole genome sequencing of the strains, and comparison of their genomic sequences to the genome of the A449 reference strain revealed various genomic rearrangements, including a new variant of the genomic island AsaGEI in BG, designated as AsaGEI2c This is the first report on a GEI of A. salmonicida strain from China. Furthermore, both YK and BG strains contained a Tn7 transposon inserted at the same position in the chromosome. Finally, IS-dependent rearrangements on pAsa5 are deemed likely to have occurred, with omission of the resD gene in both strains as well as omission of genes related to the IncF conjugal transfer system in the YK isolate. This study demonstrates that A. salmonicida subsp. salmonicida can infect non-salmonids (cyprinids) in addition to salmonids, and that AsaGEI2c might be useful as a geographical indicator of Chinese A. salmonicida subsp. salmonicida isolates.

背景与目标： : 从中国大嘴青铜鱼和北白鲑中分离出两株沙门氏菌气单胞菌YK和BG，并鉴定为沙门氏菌亚种。基于vapA和16S rRNA基因序列的系统发育分析的沙门氏菌。YK和BG起源于淡水鱼，其中一种属于鲤科，菌株表现出毒力差异。随后，我们对菌株进行了全基因组测序，并将其基因组序列与A449参考菌株的基因组进行比较，揭示了各种基因组重排，包括BG中的基因组岛AsaGEI的新变体，被指定为AsaGEI2c这是来自中国的沙门氏菌菌株GEI的第一份报告。此外，YK和BG菌株均包含插入染色体相同位置的Tn7转座子。最后，认为pAsa5上的IS依赖性重排可能已经发生，两个菌株中resD基因均被省略，并且YK分离株中与IncF结合转移系统相关的基因也被省略。这项研究表明，沙门氏菌亚种。除鲑鱼外，鲑鱼还可以感染非鲑鱼 (鲤科鱼)，而AsaGEI2c可能用作中国鲑鱼亚种的地理指标。鲑鱼分离株。

BACKGROUND & AIMS: PURPOSE:Common data elements (CDEs) are increasingly being used by researchers to promote data sharing across studies. The purposes of this article are to (a) describe the theoretical, conceptual, and definition issues in the development of a set of CDEs for research addressing self-management of chronic conditions; (b) propose an initial set of CDEs and their measures to advance the science of self-management; and (c) recommend implications for future research and dissemination. DESIGN AND METHODS:Between July 2014 and December 2015 the directors of the National Institute of Nursing Research (NINR)-funded P20 and P30 centers of excellence and NINR staff met in a series of telephone calls and a face-to-face NINR-sponsored meeting to select a set of recommended CDEs to be used in self-management research. A list of potential CDEs was developed from examination of common constructs in current self-management frameworks, as well as identification of variables frequently used in studies conducted in the centers of excellence. FINDINGS:The recommended CDEs include measures of three self-management processes: activation, self-regulation, and self-efficacy for managing chronic conditions, and one measure of a self-management outcome, global health. CONCLUSIONS:The self-management of chronic conditions, which encompasses a considerable number of processes, behaviors, and outcomes across a broad range of chronic conditions, presents several challenges in the identification of a parsimonious set of CDEs. This initial list of recommended CDEs for use in self-management research is provisional in that it is expected that over time it will be refined. Comment and recommended revisions are sought from the research and practice communities. CLINICAL RELEVANCE:The use of CDEs can facilitate generalizability of research findings across diverse population and interventions.

背景与目标：

BACKGROUND & AIMS: :To investigate the effects of air pollution related with the gasoline/petrochemical industry the expression of metallothionein I (MT-I) mRNA and tissue metals were analyzed in organs of mice, exposed to gasoline (G) vapor in laboratory conditions. Control groups consisted of intact mice and of those exposed in the metabolic chamber to fresh air. The data obtained by RT-PCR and inductively coupled plasma spectrometry have shown that exposure to G vapor leads to upregulation of MT-I mRNA in organs that receive a strong respiratory and olfactory input or participate in gasoline degradation and elimination (lungs, brain, kidney and liver). Besides, in the brain and in the lungs, kidney and liver a decreased tissue content of Zn²⁺ or Cu²⁺ and Mg²⁺ was found (p<0.001). Some of these changes were obtained also in mice closed in the metabolic chamber, pointing to the involvement of stress-induced mechanisms in the transcriptional regulation of MTs.

背景与目标： : 为了研究与汽油/石化工业有关的空气污染的影响，在实验室条件下，对暴露于汽油 (G) 蒸气的小鼠器官中金属硫蛋白I (mt-i) mRNA和组织金属的表达进行了分析。对照组由完整的小鼠和在代谢室中暴露于新鲜空气的小鼠组成。通过rt-pcr和电感耦合等离子体光谱法获得的数据表明，暴露于G蒸气会导致接受强烈呼吸和嗅觉输入或参与汽油降解和消除 (肺，脑，肾和肝) 的器官中mt-i mRNA上调。此外，在大脑和肺部，肾脏和肝脏中，发现zn ² 或cu ² 和mg ² 的组织含量降低 (p<0.001)。这些变化中的一些也在关闭在代谢室中的小鼠中获得，这表明应激诱导的机制参与了MTs的转录调控。

BACKGROUND & AIMS: :Osteocalcin and osteopontin are noncollagenous proteins secreted by osteoblasts and regulated by a complex interplay of systemic and locally produced factors, including growth factors and steroid hormones. We investigated the mechanism by which transforming growth factor-beta (TGF beta) inhibits 1,25-dihydroxyvitamin D3 (1,25-(OH)2D3)-enhanced expression of the osteocalcin (OC) and osteopontin (OP) genes. ROS 17/2.8 cells, in which both genes are expressed, were transfected with reporter constructs driven by native (i.e. wild-type) rat OC and mouse OP promoters. TGF beta abrogated the 1,25-(OH)2D3 enhanced transcription of both the OC and OP genes. The inhibitory TGF beta response for each requires vitamin D response element (VDRE) sequences, although there are additional contributions from proximal basal regulatory elements. These transcriptional effects were further investigated for contribution of the trans-activating factors, which interact with OC and OP VDREs, involving the vitamin D receptor (VDR) and retinoid X receptor (RXR). Gel mobility shift assays show that TGF beta significantly reduces induction of the heterodimers VDR/RXR complexes in 1,25-(OH)2D3-treated ROS 17/2.8 cells. However, Western blot and ligand binding analysis reveal that TGF beta does not affect nuclear availability of the VDR. We also show that activator protein-1 activity is up-regulated by TGF beta; thus, activator protein-1 binding sites in the OC promoter may potentially contribute to inhibitory effects of TGF beta on basal transcription. Our studies demonstrate that the inhibitory action of TGF beta on the 1,25-(OH)2D3 enhancement of OC and OP transcription in osteoblastic cells results from modulations of protein-DNA interactions at the OC and OP VDRE, which cannot be accounted for by changes in VDR protein levels. As OC and OP participate in bone turnover, our results provide insight into the contributions of TGF beta and 1,25-(OH)2D3 to VDR-mediated gene regulatory mechanism operative in bone formation and/or resorption events.

背景与目标： 骨钙素和骨桥蛋白是由成骨细胞分泌的非胶原蛋白，受全身和局部产生的因子 (包括生长因子和类固醇激素) 的复杂相互作用调节。我们研究了转化生长因子 β (TGF β) 抑制1,25-二羟基维生素D3 (1,25-(OH)2D3) 增强的骨钙素 (OC) 和骨桥蛋白 (OP) 基因表达的机制。用由天然 (即野生型) 大鼠OC和小鼠OP启动子驱动的报告构建体转染两个基因均表达的ROS 17/2.8细胞。TGF β 废除了1,25-(OH)2D3增强了OC和OP基因的转录。尽管近端基础调节元件还有其他贡献，但每种药物的抑制性TGF β 反应都需要维生素d反应元件 (VDRE) 序列。进一步研究了这些转录作用的反式激活因子的作用，这些反式激活因子与OC和OP VDREs相互作用，涉及维生素d受体 (VDR) 和类维生素a X受体 (RXR)。凝胶迁移率变化测定显示TGF β 显著降低在1,25-(OH) 2d3处理的ROS 17/2.8细胞中对异二聚体VDR/RXR复合物的诱导。然而，蛋白质印迹和配体结合分析表明，TGF β 不会影响VDR的核可用性。我们还显示TGF β 上调了激活蛋白1的活性; 因此，OC启动子中的激活蛋白1结合位点可能潜在地有助于TGF β 对基础转录的抑制作用。我们的研究表明，TGF β 对成骨细胞中OC和OP转录的1,25-(OH)2D3增强的抑制作用是由OC和OP VDRE处蛋白质-DNA相互作用的调节引起的，这不能解释VDR蛋白水平的变化。当OC和OP参与骨转换时，我们的结果提供了对TGF β 和1,25-(OH)2D3对VDR介导的在骨形成和/或吸收事件中起作用的基因调节机制的贡献的见解。

BACKGROUND & AIMS: :High throughput screening systems are the preferred solution to meet the urgent requirement of increasing number of genetically modified organisms (GMOs). In this study, we have successfully developed a multiplex GMO element screening system with dual priming oligonucleotide (DPO) primers. This system can detect the cauliflower mosaic virus 35S (CaMV 35S), terminator of nopaline synthase gene (NOS), figwort mosaic virus 35S (FMV 35S) promoter, neomycin phosphotransferaseII (NPTII), Bt Cry 1Ab, phosphinothricin acetyltransferase genes (bar) and Streptomyces viridochromogenes (pat) simultaneously, which covers more than 90% of all authorized GMO species worldwide. This system exhibits a high tolerance to annealing temperatures, high specificity and a limit of detection equal to conventional PCR. A total of 214 samples from markets, national entry-exit agencies, the Institute for Reference Materials and Measurement (IRMM) and the American Oil Chemists' Society (AOCS) were also tested for applicability. This screening system is therefore suitable for GMO screening.

背景与目标： : 高通量筛选系统是满足越来越多的转基因生物 (gmo) 的迫切需求的首选解决方案。在这项研究中，我们成功地开发了具有双引发寡核苷酸 (DPO) 引物的多重GMO元件筛选系统。该系统可检测花椰菜花叶病毒35S (CaMV 35S) 、胭脂树碱合酶基因终止子 (NOS) 、无花叶病毒35S (FMV 35S) 启动子、新霉素磷酸转移酶 (NPTII) 、Bt Cry 1Ab、膦丝菌素乙酰转移酶基因 (bar) 和链霉菌 (pat) 同时覆盖全球所有授权转基因物种的90% 以上。该系统表现出对退火温度的高耐受性，高特异性和与常规PCR相同的检测极限。还测试了来自市场，国家出入境机构，参考材料与测量研究所 (IRMM) 和美国石油化学家协会 (AOCS) 的总共214个样品的适用性。因此，该筛选系统适用于GMO筛选。

BACKGROUND & AIMS: :The metal accumulation potential and its physiological effects in Indian mustard plants (Brassica nigra L.) grown in soil irrigated with post methanated distillery effluent (25%, 50%, 75%, 100%, v/v) were studied after 30, 60 and 90 days after sowing. An increase in the chlorophyll and protein contents was recorded at the lower concentrations of post methanated distillery effluent (PMDE) at initial exposure periods followed by a decrease at higher concentrations of PMDE compared to their respective controls. An enhanced lipid peroxidation in tested plants was observed, which was evidenced by the increased malondialdehyde content in shoot, leaves and seeds at all the concentrations of PMDE and exposure periods compared to their respective controls. This study revealed that Indian mustard plants (B. nigra L.) are well adopted to tolerate and accumulate high quantities of trace elements due to increased level of antioxidants (cysteine and ascorbic acid) in root, shoot and leaves of the treated plants at all the concentrations and exposure periods except at 90 days, whereas a decrease was observed at 100% PMDE as compared to their respective controls.

背景与目标： : 在30、60和90天后，研究了在甲烷化蒸馏废水 (25% 、50% 、75% 、100% 、v/v) 灌溉的土壤中生长的印度芥菜植物 (Brassica nigra L.) 中的金属积累潜力及其生理效应播种后。在初始暴露期间，甲烷化蒸馏后流出物 (PMDE) 的浓度较低时，叶绿素和蛋白质含量增加，随后与各自的对照相比，在较高浓度的PMDE下降低。在测试的植物中观察到脂质过氧化作用增强，这可以通过在所有PMDE浓度和暴露时间下与各自的对照相比，芽，叶和种子中的丙二醛含量增加来证明。这项研究表明，印度芥菜植物 (B. nigra L.) 被很好地用来耐受和积累大量的微量元素，因为在处理过的植物的根，芽和叶片中抗氧化剂 (半胱氨酸和抗坏血酸) 的含量增加了，除了90天的浓度和暴露时间，而与它们各自的对照相比，在100% PMDE处观察到降低。

BACKGROUND & AIMS: :Human T lymphocyte clones with specific proliferative response to influenza A virus were derived by limiting dilution from peripheral blood lymphocytes (PBL) after in vitro stimulation with autologous irradiated, virus-infected PBL. Four OKT3+4+8- T lymphocyte clones (TLC) that showed HLA-restricted antigen-specific proliferative responses were used for a detailed analysis of the restriction elements for antigen presentation. None of the clones showed alloreactivity and all required the presence on the antigen-presenting cell of HLA class II antigens of one or other haplotype of the donor. Restriction elements for two clones were correlated with Dw1 rather than DR1, and for two others with Dw6 rather than DRw6. These latter clones showed differential recognition of HLA-Dw6 subtypes as defined tentatively by homozygous typing cells, without relationship to putative serological "splits" of DRw6. One of the Dw6-restricted clones was specific for a Dw6.1 (now Dw18) "subtype," confirmed by family segregation analysis, the other for a broad Dw6 (Dw18 and Dw19) specificity. Studies with a panel of monoclonal antibodies against monomorphic determinants of HLA class II antigens revealed heterogeneous patterns of blocking activity, distinguishing between clones of different restriction specificity. Inhibition patterns were partly as predictable from the known activity of the monoclonal antibody in alloantigeneic PLT systems. These results provide evidence that certain structures that function as restriction elements for antigen presentation also carry alloantigeneic determinants.

背景与目标： : 对甲型流感病毒具有特异性增殖反应的人T淋巴细胞克隆是通过在体外用自体辐照病毒病毒感染的PBL刺激后限制外周血淋巴细胞 (PBL) 的稀释而获得的。四个OKT3 4 8- T淋巴细胞克隆 (TLC) 显示出HLA限制性抗原特异性增殖反应，用于详细分析抗原呈递的限制性元件。所有克隆均未显示同种反应性，并且都需要在供体的一种或其他单倍型的HLA II类抗原的抗原呈递细胞上存在。两个克隆的限制元素与Dw1而不是DR1相关，另外两个克隆的限制元素与Dw6而不是drw6相关。这些后者克隆显示了由纯合型细胞初步定义的HLA-Dw6亚型的差异识别，与drw6的推定血清学 “分裂” 没有关系。Dw6-restricted克隆之一对Dw6.1 (现为Dw18) “亚型” 具有特异性，通过家庭隔离分析证实，另一个克隆具有广泛的Dw6 (Dw18和Dw19) 特异性。用一组针对HLA II类抗原单形决定簇的单克隆抗体进行的研究揭示了阻断活性的异质模式，区分了具有不同限制性特异性的克隆。抑制模式部分可根据同种异体PLT系统中单克隆抗体的已知活性预测。这些结果提供了证据，表明某些充当抗原呈递限制元件的结构也带有同种异体决定簇。

BACKGROUND & AIMS: The Drosophila s15 chorion gene is expressed only in the follicular epithelium surrounding the developing oocyte, with tight quantitative control and a very narrow temporal specificity. We have used germ-line transformation analysis to conduct an extensive mutational dissection of its promoter between -189 and -39 bp relative to the transcriptional start site. Quantitative control and temporal specificity are disrupted by several of these mutations. The results suggest that this 150-bp DNA region encompasses many positive and negative, at least partially degenerate, cis-regulatory elements, which are involved in specifying the highly precise expression pattern of the s15 gene during development.

背景与目标： 果蝇s15绒毛膜基因仅在发育中的卵母细胞周围的卵泡上皮中表达，具有严格的定量控制和非常狭窄的时间特异性。我们已经使用种系转化分析对其启动子相对于转录起始位点在-189和-39 bp之间进行了广泛的突变解剖。定量控制和时间特异性被其中几个突变破坏。结果表明，该150 bp DNA区域包含许多阳性和阴性，至少部分退化的顺式调节元件，这些元件参与确定s15基因在发育过程中的高度精确表达模式。

BACKGROUND & AIMS: Foodborne bacterial diseases cause considerable morbidity and mortality throughout the world. Preventive measures such as good manufacturing practices (GMP), supplemented by the hazard analysis critical control point (HACCP) system, have been introduced as a means of ensuring the production of safe food. However, their use does not necessarily provide quantitative information on the risks associated with the consumption of a particular food product. To obtain such information, elements of quantitative risk analysis (QRA) need to be used. QRA is defined as a stepwise analysis of the health risks associated with a specific type of food product, resulting in an estimation of the probability of occurrence of adverse effects on health following consumption of the food in question. It also includes an analysis of the nature of the risks. Taking this definition, five successive steps can be recognizedhazard identification, exposure assessment, dose response assessment, risk characterization and risk management. Food production is a dynamic activity, involving changes in, e.g. the composition and microbial quality of raw materials due to seasonal variation. Also, there may be continuing changes in processing conditions and in product composition due to consumer demands. Therefore, it will be desirable to incorporate QRA in existing safety assurance systems, such as HACCP, when sufficient information is available to permit this approach.

背景与目标： 食源性细菌性疾病在全世界引起相当大的发病率和死亡率。已引入诸如良好生产规范 (GMP) 之类的预防措施，并辅以危害分析关键控制点 (HACCP) 系统，以确保生产安全食品。但是，它们的使用不一定提供与特定食品消费相关的风险的定量信息。为了获得此类信息，需要使用定量风险分析 (QRA) 的要素。QRA被定义为对与特定类型的食品相关的健康风险的逐步分析，从而估计在食用相关食品后对健康产生不利影响的可能性。它还包括对风险性质的分析。采用此定义，可以识别危害识别，暴露评估，剂量反应评估，风险特征和风险管理五个连续步骤。食品生产是一种动态活动，涉及季节性变化，例如原材料的组成和微生物质量的变化。此外，由于消费者的需求，加工条件和产品组成可能会持续变化。因此，当有足够的信息允许这种方法时，将QRA纳入现有的安全保证系统 (例如HACCP) 中是可取的。

BACKGROUND & AIMS: :A novel murine stromal cell line, HESS-M28, was established, which supports the expansion of human CD34+CD38- cells more than 300-fold in vitro in the presence of human IL-3 and SCF. These cells were used in an attempt to evaluate cis-acting elements of retroviral vectors in human primitive hematopoietic cells. Cord blood cells were cultured on top of the mixed cell layers of the stromal cell line, HESS-M28, and retroviral vector-producing cells. The FMEV-type vector SF/Lyt contained the spleen focus-forming virus U3 and the MESV primer-binding site (PBS), while MO3/Lyt contained the U3 region and PBS from Mo-MuLV. After transduction by the FMEV-type and Mo-MuLV-based vectors, expression of the marker gene murine CD8 (mCD8) was examined in CD34-, CD34+, and CD34+CD38- cells. In CD34+ and CD34+CD38- cells, expression of mCD8 was higher with the FMEV-type vector, SF/Lyt, compared with the cells transduced by the Mo-MuLV-based vector MO3/Lyt, although the expression was comparable in CD34- cells. Expression of marker genes was also confirmed in long-term culture-initiating cells (LTC-ICs) and SCID-repopulating cells (SRCs).

背景与目标： : 建立了一种新型的鼠基质细胞系HESS-M28，该细胞系在存在人IL-3和SCF的情况下，在体外支持人CD34 CD38细胞扩增300倍以上。这些细胞用于评估人类原始造血细胞中逆转录病毒载体的顺式作用元件。在基质细胞系、HESS-M28和逆转录病毒载体产生细胞的混合细胞层的顶部培养脐带血细胞。FMEV型载体SF/Lyt包含脾脏焦点形成病毒U3和MESV引物结合位点 (PBS)，而MO3/Lyt包含来自Mo-MuLV的U3区域和PBS。通过FMEV型和基于Mo-MuLV的载体转导后，在CD34-，CD34和CD34 CD38-细胞中检查了标记基因鼠CD8 (mCD8) 的表达。在CD34和CD34 CD38-细胞中，与基于Mo-MuLV的载体MO3/Lyt转导的细胞相比，FMEV型载体SF/Lyt的mCD8表达更高，尽管其表达在CD34-细胞中相当。标记基因的表达也在长期培养起始细胞 (LTC-ICs) 和SCID再填充细胞 (src) 中得到证实。

BACKGROUND & AIMS: :Dynein interacts with microtubules through an ATP-sensitive linkage mapped to a structurally complex region of the heavy chain following the fourth P-loop motif. Virtually nothing is known regarding how binding affinity is achieved and modulated during ATP hydrolysis. We have performed a detailed dissection of the microtubule contact site, using fragment expression, alanine substitution, and peptide competition. Our work identifies three clusters of amino acids important for the physical contact with microtubules; two of these fall within a region sharing sequence homology with MAP1B, the third in a region just downstream. Amino acid substitutions within any one of these regions can eliminate or weaken microtubule binding (KK3379, 80, E3385, K3387, K3397, KK3410,11, W3414, RKK3418-20, F3426, R3464, S3466, and K3467), suggesting that their activities are highly coordinated. A peptide that actively displaces MAP1B from microtubules perturbs dynein binding, supporting previous evidence for similar sites of interaction. We have also identified four amino acids whose substitutions affect release of the motor from the microtubule (E3413, R3444, E3460, and C3469). These suggest that nucleotide-sensitive affinity may be locally controlled at the site of contact. Our work is the first detailed description of dynein-tubulin interactions and provides a framework for understanding how affinity is achieved and modulated.

背景与目标： : 动力蛋白通过ATP敏感的键与微管相互作用，该键映射到第四个P环基序后重链的结构复杂区域。关于在ATP水解过程中如何实现和调节结合亲和力，几乎一无所知。我们使用片段表达，丙氨酸取代和肽竞争对微管接触部位进行了详细的解剖。我们的工作确定了对与微管的物理接触很重要的三个氨基酸簇; 其中两个位于与MAP1B共享序列同源性的区域内，第三个位于下游区域。这些区域中任何一个区域内的氨基酸取代可以消除或削弱微管结合 (KK3379、80、E3385、K3387、K3397、KK3410、11、W3414、RKK3418-20、F3426、R3464、S3466和K3467)，表明它们的活性高度协调。一种从微管中主动置换MAP1B的肽会干扰动力蛋白的结合，支持先前的相似相互作用位点的证据。我们还鉴定了四种氨基酸，其取代会影响微管中马达的释放 (E3413，R3444，E3460和C3469)。这些表明核苷酸敏感的亲和力可能在接触部位受到局部控制。我们的工作是对动力蛋白-微管蛋白相互作用的第一个详细描述，并为理解如何实现和调节亲和力提供了框架。

BACKGROUND & AIMS: OBJECTIVE:To investigate whether the abnormal expression of inducible nitric oxide synthase (iNOS) by osteoarthritic (OA) human chondrocytes is associated with changes in the DNA methylation status in the promoter and/or enhancer elements of iNOS. METHODS:Expression of iNOS was quantified by quantitative reverse transcriptase-polymerase chain reaction. The DNA methylation status of the iNOS promoter and enhancer regions was determined by bisulfite sequencing or pyrosequencing. The effect of CpG methylation on iNOS promoter and enhancer activities was determined using a CpG-free luciferase vector and a CpG methyltransferase. Cotransfections with expression vectors encoding NF-κB subunits were carried out to analyze iNOS promoter and enhancer activities in response to changes in methylation status. RESULTS:The 1,000-bp iNOS promoter has only 7 CpG sites, 6 of which were highly methylated in both control and OA samples. The CpG site at -289 and the sites in the starting coding region were largely unmethylated in both groups. The NF-κB enhancer region at -5.8 kb was significantly demethylated in OA samples compared with control samples. This enhancer element was transactivated by cotransfection with the NF-κB subunit p65, alone or together with p50. Critically, methylation treatment of the iNOS enhancer element significantly decreased its activity in a reporter assay. CONCLUSION:These findings demonstrate the association between demethylation of specific NF-κB-responsive enhancer elements and the activation of iNOS transactivation in human OA chondrocytes, consistent with the differences in methylation status observed in vivo in normal and human OA cartilage and, importantly, show association with the OA process.

背景与目标：

BACKGROUND & AIMS: :Mobile DNA elements play a major role in genome plasticity and other evolutionary processes, an insight gained primarily through the study of transposons and retrotransposons (generally approximately 1000 nt or longer). These elements spawn smaller parasitic versions (generally >100 nt) that propagate through proteins encoded by the full elements. Highly repeated sequences smaller than 100 nt have been described, but they are either nonmobile or their origins are not known. We have surveyed the genome of the multicellular cyanobacterium, Nostoc punctiforme, and its relatives for small dispersed repeat (SDR) sequences and have identified eight families in the range of from 21 to 27 nucleotides. Three of the families (SDR4, SDR5, and SDR6), despite little sequence similarity, share a common predicted secondary structure, a conclusion supported by patterns of compensatory mutations. The SDR elements are found in a diverse set of contexts, often embedded within tandemly repeated heptameric sequences or within minitransposons. One element (SDR5) is found exclusively within instances of an octamer, HIP1, that is highly over-represented in the genomes of many cyanobacteria. Two elements (SDR1 and SDR4) often are found within copies of themselves, producing complex nested insertions. An analysis of SDR elements within cyanobacterial genomes indicate that they are essentially confined to a coherent subgroup. The evidence indicates that some of the SDR elements, probably working through RNA intermediates, have been mobile in recent evolutionary time, making them perhaps the smallest known mobile elements.

背景与目标： : 移动DNA元件在基因组可塑性和其他进化过程中起主要作用，这主要是通过研究转座子和反转录转座子 (通常约1000 nt或更长) 而获得的见解。这些元素产生较小的寄生版本 (通常> 100 nt)，其通过由完整元素编码的蛋白质传播。已经描述了小于100 nt的高度重复序列，但是它们不是可移动的，或者它们的起源是未知的。我们已经调查了多细胞蓝细菌Nostoc punctiforme的基因组及其亲属的小分散重复序列 (SDR) 序列，并确定了21至27个核苷酸范围内的8个家族。尽管序列相似性很小，但三个家族 (SDR4，SDR5和SDR6) 具有共同的预测二级结构，这一结论得到了代偿性突变模式的支持。SDR元素存在于多种环境中，通常嵌入在连续重复的七聚体序列中或在微型子中。仅在八聚体HIP1的实例中发现了一种元素 (SDR5)，该元素在许多蓝细菌的基因组中高度代表。在它们的副本中通常会发现两个元素 (SDR1和SDR4)，从而产生复杂的嵌套插入。对蓝细菌基因组中SDR元素的分析表明，它们基本上局限于一个相干的亚组。证据表明，某些SDR元素 (可能通过RNA中间体起作用) 在最近的进化时间内已经移动，这使它们成为已知最小的移动元素。

BACKGROUND & AIMS: :Trace metal elements (TME) can be real threats for living organisms. However, few studies dealt with TME in reservoirs in rural areas where farming practises could induce negative effects. Mae Thang reservoir (northern Thailand) has been studied for 3 years to understand the seasonal behaviour of dissolved TME: Fe, Mn, Cd, Al, Pb, V, Cr, Co, Ni, Cu, Zn, Mo, U and As and associated physicochemical parameters. In situ measurements of these parameters were done during the dry and the wet seasons as well as water samples along the water column for further analyses and TME determination by inductively coupled plasma-mass spectrometry (ICP-MS). In the dry season, the water column was characterized by a strong stratification and anoxic conditions in the hypolimnion. High rain and water input from the watershed during the wet season induced mixing of the water. All TME, except Ni, Co and Cr were less concentrated in the wet season indicating a dilution effect by water input. There was thus no important dissolved pollution coming from the watershed. The anoxic conditions in the dry season enhanced the reduction of Fe and Mn and the desorption processes. Depth, and thus oxic-anoxic conditions were the main drivers of TME in the dry season, while in the wet season, dissolution processes from parent rocks of watershed were favoured. The average concentrations of TME in the reservoir were in the limit of the international and Thai standards. Only localized values in the bottom of the reservoir for Fe and Mn were higher than the limits.

背景与目标： : 微量金属元素 (TME) 可能是活生物体的真正威胁。然而，很少有研究涉及农村地区水库中的TME，在这些地区，农业实践可能会产生负面影响。对Mae Thang水库 (泰国北部) 进行了3年的研究，以了解溶解的TME的季节性行为: Fe，Mn，Cd，Al，Pb，V，Cr，Co，Ni，Cu，Zn，Mo，U和As以及相关的理化参数。在干燥和潮湿季节以及沿水柱的水样中对这些参数进行了原位测量，以进一步分析和通过电感耦合等离子体质谱法 (icp-ms) 测定TME。在干旱季节，水柱的特征是hypolimnion中的强分层和缺氧条件。在雨季期间，流域的高雨水和水输入会引起水的混合。除Ni，Co和Cr外，所有TME在雨季的浓度均较低，表明水分输入具有稀释作用。因此，流域没有重要的溶解污染。干旱季节的缺氧条件增强了Fe和Mn的还原和解吸过程。在干旱季节，深度和缺氧条件是TME的主要驱动力，而在雨季，则有利于流域母岩的溶解过程。水库中TME的平均浓度在国际和泰国标准的限制内。仅储层底部的Fe和Mn局部值高于极限值。