BACKGROUND & AIMS: :The chromosome of Streptomyces coelicolor A3(2), a model organism for the genus Streptomyces, contains a cryptic type I polyketide synthase (PKS) gene cluster which was revealed when the genome was sequenced. The ca. 54-kb cluster contains three large genes, cpkA, cpkB and cpkC, encoding the PKS subunits. In silico analysis showed that the synthase consists of a loading module, five extension modules and a unique reductase as a terminal domain instead of a typical thioesterase. All acyltransferase domains are specific for a malonyl extender, and have a B-type ketoreductase. Tailoring and regulatory genes were also identified within the gene cluster. Surprisingly, some genes show high similarity to primary metabolite genes not commonly identified in any antibiotic biosynthesis cluster. Using western blot analysis with a PKS subunit (CpkC) antibody, CpkC was shown to be expressed in S. coelicolor at transition phase. Disruption of cpkC gave no obvious phenotype.

背景与目标： ：Streptomyces coelicolor A3（2）（Streptomyces属的一种模式生物）的染色体包含一个隐秘的I型聚酮化合物合酶（PKS）基因簇，该基因簇在对基因组进行测序时就可以显示出来。的ca。 54kb的簇包含三个大基因，编码PKS亚基的cpkA，cpkB和cpkC。在计算机分析中，该合成酶由一个加载模块，五个扩展模块和一个独特的还原酶（而不是典型的硫酯酶）作为末端结构域组成。所有酰基转移酶结构域都对丙二酰增量剂具有特异性，并具有B型酮还原酶。还可以在基因簇中鉴定出剪裁和调节基因。出乎意料的是，一些基因显示出与任何抗生素生物合成簇中未普遍鉴定的初级代谢产物基因高度相似。使用具有PKS亚基（CpkC）抗体的Western印迹分析，显示CpkC在过渡阶段在天蓝色链霉菌中表达。 cpkC的破坏没有明显的表型。

BACKGROUND & AIMS: CONTEXT:In animal models, estrogen inhibits atherogenesis by inhibiting many of the early steps of atherosclerotic plaque formation. However, the lack of cardioprotective effect by postmenopausal hormone replacement therapy and possible increase in cardiovascular events observed during the first year after the initiation of hormone replacement therapy may suggest that once the plaque is formed, estrogen may have additional effects that may counteract its beneficial outcomes. Indeed, the effect of estrogen on plaque stability has not been identified. OBJECTIVE:We hypothesized that 17beta-estradiol (E2) may cause increased apoptosis in human coronary artery endothelial cells (HCAECs). This effect would explain an adverse effect on plaque stability in vivo. INTERVENTION(S) AND MAIN OUTCOME MEASURE(S):The effect of E2 on apoptosis, cell proliferation, and expression of proapoptotic molecules Fas and Fas ligand (FasL) in cultured HCAECs was evaluated. RESULTS:HCAECs in culture treated with E2 showed an increase in DNA strand breaks and nuclear fragmentation indicative of apoptosis. E2 treatment also induced a significant concentration-dependent increase in Fas mRNA and protein expressions in HCAECs. Moreover, the expression of FasL mRNA and secretion of FasL protein by HCAECs were enhanced in response to E2 treatments. CONCLUSIONS:E2 increases the apoptosis in cultured HCAECs. Enhanced Fas and FasL expressions in response to E2 suggest that activation of the Fas/FasL pathway may be a mediator of the proapoptotic effects of E2 in these cells.

背景与目标： 背景：在动物模型中，雌激素通过抑制动脉粥样硬化斑块形成的许多早期步骤来抑制动脉粥样硬化。但是，绝经后激素替代疗法缺乏心脏保护作用，并且在开始激素替代疗法后的第一年内观察到的心血管事件可能增加，这表明一旦形成斑块，雌激素可能会产生额外的作用，从而抵消其有益结果。实际上，尚未确定雌激素对斑块稳定性的作用。
目的：我们假设17β-雌二醇（E2）可能导致人冠状动脉内皮细胞（HCAEC）凋亡增加。该作用将解释对体内斑块稳定性的不利影响。
干预和主要观察指标：评估了E2对培养的HCAEC中细胞凋亡，细胞增殖以及促凋亡分子Fas和FasL配体（FasL）表达的影响。
结果：用E2处理的培养物中的HCAECs显示DNA链断裂的增加和核碎裂指示凋亡。 E2处理还诱导了HCAECs Fas mRNA和蛋白表达的浓度依赖性显着增加。而且，响应于E2处理，HCAECs的FasL mRNA的表达和FasL蛋白的分泌得到增强。
结论：E2增加了培养的HCAECs的细胞凋亡。响应E2增强的Fas和FasL表达表明Fas / FasL途径的激活可能是这些细胞中E2促凋亡作用的介质。

BACKGROUND & AIMS: :6-Pyruvoyltetrahydropterin synthase (PTPS) catalyzes the second step of tetrahydrobiopterin (BH4) synthesis. We previously identified PTPS orthologs (bPTPS-Is) in bacteria which do not produce BH4. In this study we disrupted the gene encoding bPTPS-I in Synechococcus sp. PCC 7942, which produces BH4-glucoside. The mutant was normal in BH4-glucoside production, demonstrating that bPTPS-I does not participate in BH4 synthesis in vivo and bringing us a new PTPS ortholog (bPTPS-II) of a bimodular polypeptide. The recombinant Synechococcus bPTPS-II was assayed in vitro to show PTPS activity higher than human enzyme. Further computational analysis revealed the presence of mono and bimodular bPTPS-II orthologs mostly in green sulfur bacteria and cyanobacteria, respectively, which are well known for BH4-glycoside production. In summary we found new bacterial PTPS orthologs, having either a single or dual domain structure and being responsible for BH4 synthesis in vivo, thereby disclosing all the bacterial PTPS homologs.

背景与目标： ：6-丙酮酰基四氢蝶呤合成酶（PTPS）催化四氢生物蝶呤（BH4）合成的第二步。我们先前在不产生BH4的细菌中鉴定了PTPS直系同源物（bPTPS-Is）。在这项研究中，我们破坏了Synechococcus sp。中编码bPTPS-1的基因。 PCC 7942，生产BH4-葡萄糖苷。该突变体在BH4-葡萄糖苷生产中是正常的，表明bPTPS-I不参与体内BH4合成，并为我们带来了双模块多肽的新PTPS直向同源物（bPTPS-II）。在体外对重组Synechocooccus bPTPS-II进行了分析，结果表明PTPS活性高于人的酶。进一步的计算分析表明，分别在绿色硫细菌和蓝细菌中分别存在单和双模块bPTPS-II直系同源物，这对于BH4-糖苷的生产是众所周知的。总之，我们发现了新的细菌PTPS直系同源物，具有单域或双域结构并负责体内的BH4合成，从而揭示了所有细菌PTPS同源物。

BACKGROUND & AIMS: :Go/Gi coupled G-protein receptor mediated transactivation is critical in the activation of receptor tyrosine kinases (RTK). Here we show that mu opioid receptor (MOR) transactivates Flk1 and platelet-derived growth factor-beta (PDGF-beta) receptors and its agonist morphine stimulates pro-angiogenic and survival-promoting signaling in mouse retinal endothelial cells (mREC). Morphine stimulates mREC proliferation in a dose dependent fashion and promotes survival to the same extent as vascular endothelial growth factor164 (VEGF164). Morphine stimulates mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) and Akt phosphorylation in a time dependent manner like VEGF in mREC. Moreover, analogous to VEGF, morphine stimulates oncogenic signal transducer and activator of transcription 3 (STAT3) signaling. Morphine as well as VEGF-induced phospho-STAT3 and phospho-Flk1 immunoprecipitated with MOR-associated proteins. In addition morphine also stimulated MOR associated PDGF-beta receptor phosphorylation. Consistent with the relationship between VEGF and MOR we found that VEGF upregulates MOR protein and RNA expression in mREC. These data suggest that MOR associates and transactivates RTKs for Flk1 and PDGF-beta, which may have a compounding effect on angiogenic signaling in endothelium. Therefore, G-Protein coupled receptors including MOR provide novel targets to develop anti-angiogenic agents.

背景与目标： ：Go / Gi偶联的G蛋白受体介导的反式激活在受体酪氨酸激酶（RTK）的激活中至关重要。在这里，我们显示mu阿片类受体（MOR）激活Flk1和血小板衍生的生长因子-β（PDGF-β）受体，其激动剂吗啡刺激小鼠视网膜内皮细胞（mREC）促血管生成和促进生存的信号传导。吗啡以剂量依赖性方式刺激mREC增殖，并以与血管内皮生长因子164（VEGF164）相同的程度促进存活。吗啡像mREC中的VEGF一样，以时间依赖性方式刺激促分裂原活化的蛋白激酶/细胞外信号调节激酶（MAPK / ERK）和Akt磷酸化。此外，类似于VEGF，吗啡刺激致癌信号转导子和转录激活子3（STAT3）信号转导。吗啡以及VEGF诱导的磷酸化STAT3和磷酸化Flk1被MOR相关蛋白免疫沉淀。此外，吗啡还刺激了MOR相关的PDGF-β受体的磷酸化。与VEGF和MOR之间的关系一致，我们发现VEGF上调mREC中的MOR蛋白和RNA表达。这些数据表明，MOR与Flk1和PDGF-beta的RTK缔合并反激活，这可能对内皮中的血管生成信号具有复合作用。因此，包括MOR在内的G蛋白偶联受体为开发抗血管生成剂提供了新的靶标。

BACKGROUND & AIMS: The stability and activity of indole-3-glycerol phosphate synthase from Sulfolobus solfataricus were studied as a function of pH and temperature. In this paper we focus on three points(1) the long-term stability of the protein to irreversible denaturation at high temperature; (2) the short-term stability of the protein to reversible temperature-driven unfolding; and (3) the dependence of its activity on temperature.

Results can be summarized as follows(a) the same first-order kinetic constant (0.020+/-0.003 min-1) was determined at different pH values (6.5, 8.0 and 9.5) from long-term stability experiments at 80 degrees C; (b) short-term stability experiments revealed different behaviour in two different pH ranges (6.5-8.0, 8.5-9.5), suggesting that the melting temperature is higher at alkaline than at neutral pH; (c) the dependence of activity on temperature was investigated at pH 7.0 and 9.0, and a discontinuity was observed in the Arrhenius plot of kcat values at pH 9.0. We also investigated the stability in the presence of guanidinium chloride at 20 degrees C either at pH 7.0 or at pH 9.0, and we present data that indicate that the unfolding mechanism closely approaches a two-state model at pH 7.0 and a more complex mechanism at pH 9.0. Satisfactory fitting of the equilibrium unfolding transition obtained by fluorescence measurements at pH 9.0 required a model that involves a stable intermediate in addition to the native and unfolded forms. At 20 degrees C the folded conformation is more stable than the unfolded conformation by (14. 7+/-1.2) kJ/mol at pH 7.0 and by (25.5+/-1.8) kJ/mol at pH 9.0.

背景与目标： 研究了来自Sulfolobus solfataricus的吲哚-3-甘油磷酸合酶的稳定性和活性随pH和温度的变化。在本文中，我们集中在三个方面：（1）蛋白质在高温下对不可逆变性的长期稳定性； （2）蛋白质对温度驱动的可逆展开的短期稳定性； （3）其活性对温度的依赖性。结果总结如下：（a）在不同的pH值下测定相同的一阶动力学常数（0.020 /-0.003 min-1）。 6.5、8.0和9.5）在80摄氏度下进行的长期稳定性实验； （b）短期稳定性实验表明，在两个不同的pH范围（6.5-8.0，8.5-9.5）中有不同的行为，这表明碱性下的熔化温度高于中性pH下的熔化温度； （c）在pH 7.0和9.0下研究了活性对温度的依赖性，并且在pH 9.0下在kcat值的Arrhenius图中观察到不连续性。我们还研究了在20摄氏度pH 7.0或pH 9.0下存在氯化胍时的稳定性，并且我们提供的数据表明，展开机理在pH 7.0时接近于两态模型，而在pH 7.0时则更复杂。 pH值9.0。通过在pH 9.0下进行荧光测量获得的平衡展开过渡的满意拟合，需要一个模型，该模型除了天然形式和未折叠形式外，还涉及稳定的中间体。在20摄氏度时，折叠构象比未折叠构象更稳定，在pH 7.0时为（14. 7 /-1.2）kJ / mol，在pH 9.0时为（25.5 /-1.8）kJ / mol。

BACKGROUND & AIMS: :A part of the new biosynthetic gene cluster for 2-deoxystreptamine-containing antibiotics was identified from Streptoalloteichus hindustanus. The alloH gene in the gene cluster was deduced to encode 2-deoxy-scyllo-inosose synthase and the expressed protein AlloH was confirmed to have this enzyme activity. Furthermore, biochemical properties of AlloH were studied.

背景与目标： ：从印度链球菌中鉴定出了一种新的生物合成基因簇，用于含有2-脱氧链胺胺的抗生素。推导了基因簇中的alloH基因编码2-脱氧-鞘氨醇合酶，并证实表达的蛋白AlloH具有这种酶活性。此外，研究了AlloH的生化特性。

BACKGROUND & AIMS: :The anaerobic metabolism of phenol proceeds via carboxylation to 4-hydroxybenzoate by a two-step process involving seven proteins and two enzymes ("biological Kolbe-Schmitt carboxylation"). MgATP-dependent phosphorylation of phenol catalyzed by phenylphosphate synthase is followed by phenylphosphate carboxylation. Phenylphosphate synthase shows similarities to phosphoenolpyruvate (PEP) synthase and was studied for the bacterium Thauera aromatica. It consists of three proteins and transfers the beta-phosphoryl from ATP to phenol; the products are phenylphosphate, AMP, and phosphate. We showed that protein 1 becomes phosphorylated in the course of the reaction cycle by [beta-(32)P]ATP. This reaction requires protein 2 and is severalfold stimulated by protein 3. Stimulation of the reaction by 1 M sucrose is probably due to stabilization of the protein(s). Phosphorylated protein 1 transfers the phosphoryl group to phenolic substrates. The primary structure of protein 1 was analyzed by nanoelectrospray mass spectrometry after CNBr cleavage, trypsin digestion, and online high-pressure liquid chromatography at alkaline pH. His-569 was identified as the phosphorylated amino acid. We propose a catalytic ping-pong mechanism similar to that of PEP synthase. First, a diphosphoryl group is transferred to His-569 in protein 1, from which phosphate is cleaved to render the reaction unidirectional. Histidine phosphate subsequently serves as the actual phosphorylation agent.

背景与目标： ：苯酚的厌氧代谢通过包含7种蛋白质和2种酶的两步过程通过羧化反应生成4-羟基苯甲酸酯（“生物学Kolbe-Schmitt羧化反应”）。苯磷酸合酶催化苯酚的MgATP依赖性磷酸化，然后苯磷酸羧化。苯磷酸合酶显示与磷酸烯醇丙酮酸（PEP）合酶的相似性，并已针对芳香族拟南芥细菌进行了研究。它由三种蛋白质组成，将β-磷酰基从ATP转移到苯酚。产品是苯基磷酸酯，AMP和磷酸酯。我们显示蛋白质1在反应周期的过程中被[β-（32）P] ATP磷酸化。该反应需要蛋白质2，并被蛋白质3刺激几倍。1 M蔗糖刺激反应可能是由于蛋白质的稳定化所致。磷酸化的蛋白1将磷酸基团转移到酚类底物上。在碱性pH下，CNBr裂解，胰蛋白酶消化和在线高压液相色谱分析后，通过纳米电喷雾质谱分析蛋白质1的一级结构。 His-569被鉴定为磷酸化氨基酸。我们提出了一种类似于PEP合酶的催化乒乓机制。首先，二磷酰基被转移到蛋白质1中的His-569上，从中裂解磷酸盐使反应单向进行。磷酸组氨酸随后用作实际的磷酸化剂。

BACKGROUND & AIMS: :Over the past decade, the old idea that the bone marrow contains endothelial cell precursors has become an area of renewed interest. While some still believe that there are no endothelial precursors in the blood, even among those who do, there is no consensus as to what they are or what they do. In this review, we describe the problems in identifying endothelial cells and conclude that expression of endothelial nitric oxide synthase may be the most reliable antigenic indicator of the phenotype. The evidence for two different classes of endothelial precursors is also presented. We suggest that, though there is no single endothelial cell precursor, we may be able to use these phenotypic variations to our advantage in better understanding their biology. We also discuss how a variety of genetic, epigenetic, and methodological differences can account for the seemingly contradictory findings on the physiological relevance of bone marrow-derived precursors in normal vascular maintenance and in response to injury. Data on the impact of tumor type and location on the contribution of bone marrow-derived cells to the tumor vasculature are also presented. These data provide hope that we may ultimately be able to predict those tumors in which bone marrow-derived cells will have a significant contribution and design therapies accordingly. Finally, factors that regulate bone marrow cell recruitment to and function in the endothelium are beginning to be identified, and several of these, including stromal derived factor 1, monocyte chemoattractant factor-1, and vascular endothelial growth factor are discussed.

背景与目标： ：在过去的十年中，关于骨髓中含有内皮细胞前体的古老观念已经引起人们的广泛关注。尽管有些人仍然认为血液中没有内皮前体，即使在有血液的人中也没有，但是关于它们是什么或它们的行为尚无共识。在这篇综述中，我们描述了鉴定内皮细胞的问题，并得出结论，内皮一氧化氮合酶的表达可能是最可靠的表型抗原指示剂。还提供了两种不同类别的内皮前体的证据。我们建议，尽管没有单个内皮细胞前体，但我们也许能够利用这些表型变异来更好地了解它们的生物学特性。我们还将讨论各种遗传，表观遗传学和方法学上的差异如何解释在正常血管维持和对损伤的反应中，骨髓来源的前体的生理相关性看似矛盾的发现。还提供了有关肿瘤类型和位置对骨髓衍生细胞对肿瘤脉管系统的影响的数据。这些数据提供了希望，使我们最终能够预测那些骨髓来源的细胞将发挥重要作用的肿瘤，并据此设计治疗方法。最后，已开始确定调节骨髓细胞向内皮募集并在内皮中起作用的因子，并讨论了其中的几种，包括基质衍生因子1，单核细胞趋化因子-1和血管内皮生长因子。

BACKGROUND & AIMS: OBJECTIVE:To compare prostate carcinoma, with and with no lymph node metastasis, to benign prostatic hyperplasia (BPH) tissue for lymphatic vessel density (LVD) and the expression of the lymph-endothelial specific growth factor, vascular endothelial growth factor C (VEGF-C), to determine their role in lymphogenic metastasis. PATIENTS, MATERIALS AND METHODS:Lymphatic vessels were stained using lymphatic vessel endothelial hyaluronan receptor 1 and assessed in standard areas. The expression of VEGF-C was assessed by the number of positive epithelial cells. The data were compared with the clinical staging. RESULTS:The lowest LVD was found in tumorous areas as opposed to periphery and nontumorous tissue (P = 0.007; P < 0.001). The highest LVD was in BPH tissue (P < 0.001). There was no correlation with clinical staging. There was more VEGF-C staining in pN1 than in pN0 and in BPH specimens (P = 0.002). CONCLUSION:LVD is not a prognostic variable for the process of lymphogenic metastasis in prostate cancer. VEGF-C is up-regulated in prostate cancer and its correlation with lymph node status suggests a role for the development of lymph node metastasis, e.g. via an increased permeability of lymphatic vessels.

背景与目标： 目的：比较有无淋巴结转移的前列腺癌与良性前列腺增生（BPH）组织的淋巴管密度（LVD）以及淋巴内皮特异性生长因子，血管内皮生长因子C（VEGF- C），确定其在淋巴转移中的作用。
患者，材料和方法：使用淋巴管内皮透明质酸受体1对淋巴管进行染色，并在标准区域进行评估。通过阳性上皮细胞的数量评估VEGF-C的表达。将数据与临床分期进行比较。
结果：与周围和非肿瘤组织相比，在肿瘤区域发现的LVD最低（P = 0.007； P <0.001）。 LVD最高的是BPH组织（P <0.001）。与临床分期无关。 pN1和BPH标本中的VEGF-C染色要多于pN0和PPH（P = 0.002）。
结论：LVD不是前列腺癌淋巴转移的预后变量。 VEGF-C在前列腺癌中被上调，并且其与淋巴结状态的相关性暗示了淋巴结转移的发展，例如肝癌的发生。通过增加淋巴管的通透性

BACKGROUND & AIMS: :The band alignment at an Al2O3/SrTiO3 heterointerface forming a two-dimensional electron gas (2DEG) was investigated using scanning photocurrent microscopy (SPCM) in an electrolyte-gated environment. We used a focused UV laser source for above-the-bandgap illumination on the SrTiO3 layer, creating electron-hole pairs that contributed to the photocurrent through migration towards the metal electrodes. The polarity of the SPCM signals of a bare SrTiO3 device shows typical p-type behavior at zero gate bias, in which the photogenerated electrons are collected by the electrodes. In contrast, the SPCM polarity of 2DEG device indicates that the hole carriers were collected by the metal electrodes. Careful transport measurements revealed that the gate-dependent conductance of the 2DEG devices exhibits n-type switching behavior. More importantly, the SPCM signals in 2DEG devices demonstrated very unique gate-responses that cannot be found in conventional semiconducting devices, based on which we were able to perform detailed investigation into the electronic band alignment of the 2DEG devices and obtain the valence band offset at the heterointerface.

背景与目标： ：使用扫描光电流显微镜（SPCM）在电解质门控环境中研究了形成二维电子气（2DEG）的Al2O3 / SrTiO3异质界面处的能带排列。我们使用聚焦的UV激光源在SrTiO3层上进行带隙以上照明，从而创建电子-空穴对，这些电子-空穴对通过向金属电极的迁移而有助于光电流。裸露的SrTiO3器件的SPCM信号的极性在零栅极偏置下显示出典型的p型行为，其中光生电子被电极收集。相反，2DEG器件的SPCM极性表明空穴载流子被金属电极收集。仔细的传输测量表明，2DEG器件的依赖于栅极的电导表现出n型开关行为。更重要的是，2DEG器件中的SPCM信号表现出非常独特的门响应，这是常规半导体器件中找不到的，基于此，我们能够对2DEG器件的电子能带对准进行详细研究，并获得价带偏移。异类接口。

BACKGROUND & AIMS: :With recent advances in synthetic biology, CO2 could be utilized as a carbon feedstock by native or engineered organisms, assuming the availability of electrons. Two key enzymes used in autotrophic CO2 fixation are the CO dehydrogenase (CODH) and acetyl coenzyme A (acetyl-CoA) synthase (ACS), which form a bifunctional heterotetrameric complex. The CODH/ACS complex can reversibly catalyze CO2 to CO, effectively enabling a biological water-gas shift reaction at ambient temperatures and pressures. The CODH/ACS complex is part of the Wood-Ljungdahl pathway (WLP) used by acetogens to fix CO2, and it has been well characterized in native hosts. So far, only a few recombinant CODH/ACS complexes have been expressed in heterologous hosts, none of which demonstrated in vivo CO2 reduction. Here, functional expression of the Clostridium carboxidivorans CODH/ACS complex is demonstrated in the solventogen Clostridium acetobutylicum, which was engineered to express CODH alone or together with the ACS. Both strains exhibited CO2 reduction and CO oxidation activities. The CODH reactions were interrogated using isotopic labeling, thus verifying that CO was a direct product of CO2 reduction, and vice versa. CODH apparently uses a native C. acetobutylicum ferredoxin as an electron carrier for CO2 reduction. Heterologous CODH activity depended on actively growing cells and required the addition of nickel, which is inserted into CODH without the need to express the native Ni insertase protein. Increasing CO concentrations in the gas phase inhibited CODH activity and altered the metabolite profile of the CODH-expressing cells. This work provides the foundation for engineering a complete and functional WLP in nonnative host organisms.IMPORTANCE Functional expression of CO dehydrogenase (CODH) from Clostridium carboxidivorans was demonstrated in C. acetobutylicum, which is natively incapable of CO2 fixation. The expression of CODH, alone or together with the C. carboxidivorans acetyl-CoA synthase (ACS), enabled C. acetobutylicum to catalyze both CO2 reduction and CO oxidation. Importantly, CODH exhibited activity in both the presence and absence of ACS. 13C-tracer studies confirmed that the engineered C. acetobutylicum strains can reduce CO2 to CO and oxidize CO during growth on glucose.

背景与目标： ：随着合成生物学的最新进展，假设电子的可用性，CO2可以被天然或工程有机体用作碳原料。自养性CO2固定中使用的两个关键酶是CO脱氢酶（CODH）和乙酰辅酶A（乙酰-CoA）合酶（ACS），它们形成了双功能异四聚体复合物。 CODH / ACS复合物可以可逆地将CO2催化转化为CO，从而有效地实现了在环境温度和压力下进行生物水煤气变换反应。 CODH / ACS复合物是乙酸原用于固定CO2的Wood-Ljungdahl途径（WLP）的一部分，并且在天然宿主中已得到很好的表征。迄今为止，在异源宿主中仅表达了少数重组CODH / ACS复合物，但均未显示体内CO2减少。在此，在溶剂原丙酮丁醇梭菌（Clostridium acetobutylicum）中证明了碳氧化梭菌CODH / ACS复合物的功能表达，该溶剂被改造成单独或与ACS一起表达CODH。两种菌株均表现出CO 2还原和CO氧化活性。使用同位素标记对CODH反应进行了询问，从而验证了CO是CO2还原的直接产物，反之亦然。 CODH显然使用天然的丙酮丁醇梭菌铁氧还蛋白作为电子载体来减少CO2。异源CODH活性取决于活跃生长的细胞，并需要添加镍，而无需表达天然Ni插入酶蛋白即可将其插入CODH。气相中CO浓度的增加会抑制CODH活性并改变表达CODH的细胞的代谢产物谱。这项工作为在非天然宿主生物中工程化完整且功能性的WLP提供了基础。重要提示在丙酮丁醇梭菌中证明了碳氧化梭菌中CO脱氢酶（CODH）的功能性表达，这本来就无法进行CO2固定。 CODH的表达，单独或与碳氧化单胞菌乙酰辅酶A合酶（ACS）一起表达，可使丙酮丁醇梭菌既催化CO2还原又催化CO氧化。重要的是，在有或没有ACS的情况下，CODH均具有活性。 13C示踪剂研究证实，工程改造的丙酮丁醇梭菌菌株可以在葡萄糖生长期间将CO2还原为CO，并氧化CO。

BACKGROUND & AIMS: :The objective of the study was to evaluate the association between peripheral venous disease (PVD) and arterial endothelial dysfunction (ED). Arterial and venous diseases have been always considered as two completely different entities, but the recent discovery of a relationship between arterial and venous thrombosis have challenged this assumption. ED, considered to be an early process in the pathophysiology of atherosclerotic disease, could represent a common pathogenetic background. We studied 39 healthy volunteers (median age: 34 years; men: 25.6%). PVD was diagnosed using ultrasound examination, arterial ED using flow-mediated dilation (FMD) and FMD normalized for the peak shear rate (nFMD). Compared with controls, participants with PVD had a lower FMD (15.2 versus 23.4%, P < 0.001) and nFMD (12.7 × 10(-3) versus 19 × 10(-3)/second, P < 0.001). People with the most clinically evident disease had the worst endothelial function. In conclusion, our findings, if confirmed in larger population, might corroborate the idea that venous and arterial disease could have common causes.

背景与目标： ：该研究的目的是评估周围静脉疾病（PVD）与动脉内皮功能障碍（ED）之间的关联。动脉和静脉疾病一直被认为是两个完全不同的实体，但是最近发现动脉和静脉血栓形成之间的关系挑战了这一假设。 ED被认为是动脉粥样硬化疾病病理生理的早期过程，可能代表了常见的致病背景。我们研究了39名健康志愿者（中位年龄：34岁；男性：25.6％）。使用超声检查诊断PVD，使用流介导的扩张（FMD）诊断动脉ED，并针对峰剪切速率（nFMD）归一化FMD。与对照组相比，PVD参与者的FMD较低（15.2比23.4％，P <0.001）和nFMD（12.7×10（-3）/ 19×10（-3）/秒，P <0.001）。临床上最明显的疾病的人的内皮功能最差。总之，我们的发现，如果在更大的人群中得到证实，可能证实静脉和动脉疾病可能是常见原因的观点。

BACKGROUND & AIMS: PURPOSE:The issue of unidentified volume expansion is well recognized as a cause for resistance to antihypertensive therapy. The aim of study is to identify contribution of negative fluid balance to hypertension control and impact on endothelial and cardiac functions among primary hypertensive patients who do not have kidney failure. MATERIALS AND METHODS:This is a prospective interventional study with one-year follow-up. Preceded by volume status measurements were performed by a body composition monitor (BCM), the patients were put on ambulatory blood pressure monitoring for 24 hours. Then, echocardiographic assessments and flow-mediated dilation (FMD) and carotid intima-media thickness (CIMT) measurements were completed. Patients in one of the two groups were kept negative hydrated during trial with diuretic treatment. RESULTS:At the end of one-year follow-up, patients in negative hydrated group were found to have significantly lower CIMT, left ventricle mass index, left ventricular end-diastolic diameter, mean systolic and diastolic BP, non-dipper patient ratio, and higher FMD. In negatively hydrated group, target organ damage significantly reduced during trial. CONCLUSIONS:The significance of negative hydration status with respect to blood pressure control, endothelial and cardiac functions within primary hypertensive patients who do not suffer from kidney failure has been demonstrated.

背景与目标： 目的：不明原因的体积膨胀问题已被公认为是抗高血压治疗耐药的原因。研究的目的是在没有肾衰竭的原发性高血压患者中确定负流体平衡对高血压控制的贡献以及对内皮和心脏功能的影响。
材料与方法：这是一项为期一年的随访的前瞻性干预研究。在通过身体成分监测仪（BCM）进行体积状态测量之前，将患者进行动态血压监测24小时。然后，完成了超声心动图评估以及血流介导的扩张（FMD）和颈动脉内膜中层厚度（CIMT）的测量。两组中的一组患者在利尿剂治疗试验期间保持负水分状态。
结果：在一年的随访结束时，负水合组患者的CIMT，左心室质量指数，左心室舒张末期直径，平均收缩压和舒张压，非北斗七星患者比率显着降低，和更高的FMD。在负水合组中，试验期间靶器官损伤显着减少。
结论：在没有肾脏衰竭的原发性高血压患者中，负水合状态对血压控制，内皮和心脏功能的重要性已得到证实。

BACKGROUND & AIMS: :Human coronary artery endothelial cells (HCAECs) have the potential to undergo fibrogenic endothelial-mesenchymal transition (EndMT), which results in matrix-producing fibroblasts and thereby contributes to the pathogenesis of cardiac fibrosis. Recently, the profibrotic cytokine transforming growth factor-β (TGF-β) is shown to be the crucial pathogenic driver which has been verified to induce EndMT. C-Ski is an important regulator of TGF-β signaling. However, the detailed role of c-Ski and the molecular mechanisms by which c-Ski affects TGF-β-induced EndMT in HCAECs are not largely elucidated. In the present study, we treated HCAECs with TGF-β of different concentrations to induce EndMT. We found that overexpression of c-Ski in HCAECs either blocked EndMT via hindering Vimentin, Snail, Slug, and Twist expression while enhancing CD31 expression, with or without TGF-β treatment. In contrast, suppression of c-Ski further enhanced EndMT. Currently, miRNA expression disorder has been frequently reported associating with cardiac fibrosis. By using online tools, we regarded miR-155 as a candidate miRNA that could target c-Ski, which was verified using luciferase assays. C-Ski expression was negatively regulated by miR-155. TGF-β-induced EndMT was inhibited by miR-155 silence; the effect of TGF-β on Vimentin, CD31, Snail, Slug, and Twist could be partially restored by miR-155. Altogether, these findings will shed light on the role and mechanism by which miR-155 regulates TGF-β-induced HCAECs EndMT via c-Ski to affect cardiac fibrosis, and miR-155/c-Ski may represent novel biomarkers and therapeutic targets in the treatment of cardiac fibrosis.

背景与目标： ：人冠状动脉内皮细胞（HCAEC）有可能经历纤维化内皮-间充质转变（EndMT），这会导致产生基质的成纤维细胞，从而促进心脏纤维化的发病机理。最近，已证明原纤维化细胞因子转化生长因子-β（TGF-β）是关键的致病性驱动因子，已被证实可诱导EndMT。 C-Ski是TGF-β信号传导的重要调节剂。然而，在很大程度上没有阐明c-Ski的详细作用以及c-Ski影响HCAECs中TGF-β诱导的EndMT的分子机制。在本研究中，我们用不同浓度的TGF-β处理HCAEC，以诱导EndMT。我们发现，无论是否使用TGF-β处理，HCAEC中c-Ski的过表达要么通过阻碍波形蛋白，蜗牛，Slug和Twist的表达而阻断EndMT，同时增强CD31的表达。相反，抑制c-Ski进一步增强了EndMT。当前，miRNA表达障碍经常被报道与心脏纤维化有关。通过使用在线工具，我们将miR-155视为可以靶向c-Ski的候选miRNA，这已通过荧光素酶测定法进行了验证。 C-Ski表达受到miR-155的负调控。 TGF-β诱导的EndMT被miR-155沉默抑制； TGF-β对波形蛋白，CD31，蜗牛，Slug和Twist的作用可以被miR-155部分恢复。总而言之，这些发现将阐明miR-155通过c-Ski调节TGF-β诱导的HCAECs EndMT通过影响心脏纤维化的作用和机制，而miR-155 / c-Ski可能代表了新型的生物标志物和治疗靶点。心脏纤维化的治疗。

BACKGROUND & AIMS: :Maintenance of the lipid composition is important for proper function and homeostasis of the mitochondrion. In Trypanosoma brucei, the enzymes involved in the biosynthesis of the mitochondrial phospholipid, phosphatidylglycerol (PG), have not been studied experimentally. We now report the characterization of T. brucei phosphatidylglycerophosphate synthase (TbPgps), the rate-limiting enzyme in PG formation, which was identified based on its homology to other eukaryotic Pgps. Lipid quantification and metabolic labelling experiments show that TbPgps gene knock-down results in loss of PG and a reduction of another mitochondria-specific phospholipid, cardiolipin. Using immunohistochemistry and immunoblotting of digitonin-isolated mitochondria, we show that TbPgps localizes to the mitochondrion. Moreover, reduced TbPgps expression in T. brucei procyclic forms leads to alterations in mitochondrial morphology, reduction in the amounts of respiratory complexes III and IV and, ultimately, parasite death. Using native polyacrylamide gel electrophoresis we demonstrate for the first time in a eukaryotic organism that TbPgps is a component of a 720 kDa protein complex, co-migrating with T. brucei cardiolipin synthase and cytochrome c1, a protein of respiratory complex III.

背景与目标： ：脂质成分的维护对于线粒体的正常功能和体内平衡非常重要。在布鲁氏锥虫中，尚未对线粒体磷脂，磷脂酰甘油（PG）的生物合成中涉及的酶进行研究。现在，我们报告T.brucei磷脂酰甘油磷酸合酶（TbPgps）的表征，PG形成中的限速酶，是根据它与其他真核Pgps的同源性鉴定的。脂质定量和代谢标记实验表明，TbPgps基因敲低会导致PG丢失并降低另一种线粒体特异性磷脂心磷脂。使用免疫组化和指印蛋白分离的线粒体的免疫印迹，我们显示TbPgps定位于线粒体。此外，布鲁氏杆菌前环形式的TbPgps表达降低导致线粒体形态改变，呼吸道复合物III和IV数量减少，最终导致寄生虫死亡。使用天然聚丙烯酰胺凝胶电泳，我们首次在真核生物中证明了TbPgps是720 kDa蛋白复合物的一个成分，与T. brucei心磷脂合酶和细胞色素c1（呼吸复合物III的蛋白）共同迁移。