BACKGROUND & AIMS: Interleukin-1 alpha (IL-1 alpha) induces cell motility in a variety of benign cell types and in some but not all malignant cell lines in vitro. This study characterizes the IL-1 alpha-induced motility of an aggressive human melanoma cell line that expresses both type I and type II IL-1 receptors. We tested the effect of monoclonal antibodies including function-blocking moAbs against the type I and type II IL-1 receptors on melanoma cell motility to determine which receptor is involved in signal transduction of IL-1 alpha-induced melanoma cell motility. IL-1 alpha significantly increases MM-RU melanoma cell migration in a dose-dependent manner using modified Boyden chamber assays at concentrations 10 to 100 times less than concentrations that significantly inhibit cell growth. Computer-assisted time-lapse image analysis reveals that the motility is inhibited in a dose-dependent manner by neutralizing antibodies against IL-1 alpha. Function-blocking monoclonal antibodies against either type I or type II IL-1 receptors show a significant inhibition of cytokine-induced enhanced cell migration. When both the anti-IL-1 receptor antibodies are added together, the motility-response is completely blocked to control levels. Taken together the data indicate that the IL-1 alpha-induced motility of MM-RU melanoma cells is mediated through both type I and type II IL-1 receptors. The significant inhibition of motility by neutralizing IL-1 alpha or blocking either one or both of the IL-1 receptors indicates an integration of IL-1-induced signals in the induction of melanoma cell migration.

背景与目标： 白细胞介素-1（IL-1 alpha）在多种良性细胞类型中以及某些但不是全部恶性细胞系中诱导细胞运动。这项研究的特点是表达I型和II型IL-1受体的侵略性人黑素瘤细胞系的IL-1α诱导的运动。我们测试了包括针对I型和II型IL-1受体的功能阻断性单抗的单克隆抗体对黑素瘤细胞运动性的影响，以确定哪个受体参与了IL-1α诱导的黑素瘤细胞运动性的信号转导。 IL-1α使用改良的Boyden室测定法以剂量依赖性方式显着增加MM-RU黑色素瘤细胞迁移，其浓度比明显抑制细胞生长的浓度低10至100倍。计算机辅助的延时图像分析表明，通过中和针对IL-1α的抗体，可以以剂量依赖性的方式抑制运动性。针对I型或II型IL-1受体的功能阻断性单克隆抗体显示出对细胞因子诱导的细胞迁移增强的显着抑制作用。当两种抗IL-1受体抗体一起添加时，运动反应完全被阻断至对照水平。数据合计表明，IL-1α诱导的MM-RU黑色素瘤细胞的运动是通过I型和II型IL-1受体介导的。通过中和IL-1α或阻断任何一个IL-1受体或两个IL-1受体来显着抑制运动性，这表明在黑素瘤细胞迁移的诱导中整合了IL-1诱导的信号。

BACKGROUND & AIMS: :Matrix assembly and homeostasis in collagen-rich tissues are mediated by interactions with proteoglycans (PGs) substituted with sulfated glycosaminoglycans (GAGs). The major GAG in cornea is keratan sulfate (KS), which is N-linked to one of three PG core proteins. To ascertain the importance of the carbohydrate chain sulfation step in KS functionality, we generated a strain of mice with a targeted gene deletion in Chst5, which encodes an N-acetylglucosamine-6-O-sulfotransferase that is integral to the sulfation of KS chains. Corneas of homozygous mutants were significantly thinner than those of WT or heterozygous mice. They lacked high-sulfated KS, but contained the core protein of the major corneal KSPG, lumican. Histochemically stained KSPGs coassociated with fibrillar collagen in WT corneas, but were not identified in the Chst5-null tissue. Conversely, abnormally large chondroitin sulfate/dermatan sulfate PG complexes were abundant throughout the Chst5-deficient cornea, indicating an alteration of controlled PG production in the mutant cornea. The corneal stroma of the Chst5-null mouse exhibited widespread structural alterations in collagen fibrillar architecture, including decreased interfibrillar spacing and a more spatially disorganized collagen array. The enzymatic sulfation of KS GAG chains is thus identified as a key requirement for PG biosynthesis and collagen matrix organization.

背景与目标： ：胶原蛋白丰富的组织中的基质组装和体内平衡是通过与被硫酸化的糖胺聚糖（GAG）取代的蛋白聚糖（PG）的相互作用而介导的。角膜中的主要GAG是硫酸角质素（KS），它与三种PG核心蛋白之一进行N-连接。为了确定碳水化合物链硫酸化步骤在KS功能中的重要性，我们生成了在Chst5中具有靶向基因缺失的小鼠品系，该菌株编码一个N-乙酰氨基葡糖-6-O-磺基转移酶，该酶是KS链硫酸化所必需的。纯合突变体的角膜明显比野生型或杂合小鼠的角膜薄。他们缺乏高硫酸盐化的KS，但含有主要角膜KSPG的核心蛋白lumican。组织化学染色的KSPG与野生型角膜中的原纤维胶原蛋白相关联，但在Chst5无组织中未发现。相反，在整个Chst5缺陷型角膜中，异常大的硫酸软骨素/硫酸皮肤素PG复合物丰富，表明突变体角膜中受控PG产生的改变。 Chst5-null小鼠的角膜基质在胶原纤维结构中表现出广泛的结构变化，包括减小的纤维间间距和空间上更混乱的胶原阵列。因此，KS GAG链的酶促硫酸化被确定为PG生物合成和胶原基质组织的关键要求。

BACKGROUND & AIMS: :Crohn's disease (CD) and ulcerative colitis (UC) are chronic inflammatory disorders of the gastrointestinal tract that share clinical and pathological characteristics. The most accredited hypothesis is that both CD and UC result from a deregulated mucosal immune response to normal constituents of the gut microflora. Evidence, however, indicates that the main pathological processes in these two diseases are distinct. In CD, the tissue-damaging inflammatory reaction is driven by activated type 1 helper T-cell (Th1), whereas a humoral response predominates in UC. Consistently, a marked accumulation of macrophages making interleukin (IL)-12, the major Th1-inducing factor, is seen in CD but not in UC mucosa. Preliminary studies also indicate that administration of a monoclonal antibody blocking the IL-12/p40 subunit can be useful to induce and maintain clinical remission in CD patients. Notably, the recently described IL-23 shares the p40 subunit with IL-12, raising the possibility that the clinical benefit of the anti-IL-12/p40 antibody in CD may also be due to the neutralization of IL-23 activity. This review summarizes the current information on the expression and functional role of IL-12 and IL-12-associated signaling pathways both in patients with CD and experimental models of colitis, thus emphasizing major differences between IL-12 and IL-23 activity on the development of intestinal inflammation.

背景与目标： ：克罗恩病（CD）和溃疡性结肠炎（UC）是胃肠道的慢性炎性疾病，具有临床和病理学特征。最公认的假设是CD和UC均来自对肠道菌群正常成分的粘膜免疫反应失控。然而，证据表明这两种疾病的主要病理过程是不同的。在CD中，组织破坏性炎症反应是由激活的1型辅助性T细胞（Th1）驱动的，而体液反应在UC中占主导地位。一致地，在CD中可见大量的巨噬细胞积聚，使白介素（IL）-12（主要的Th1诱导因子）在CD中可见，而在UC粘膜中则未见。初步研究还表明，给予阻断IL-12 / p40亚基的单克隆抗体可用于诱导和维持CD患者的临床缓解。值得注意的是，最近描述的IL-23与IL-12共有p40亚基，从而增加了抗IL-12 / p40抗体在CD中的临床益处也可能是由于IL-23活性的中和所引起的可能性。这篇综述总结了有关CD患者和结肠炎实验模型中IL-12和IL-12相关信号通路的表达和功能作用的最新信息，从而强调了IL-12和IL-23活性在结肠炎患者中的主要差异。肠道炎症的发展。

BACKGROUND & AIMS: :The aim of this study was to examine the different patterns of cerebellar and/or brainstem atrophy in spinocerebellar ataxia (SCA) type 3 and 6. Eighteen patients (SCA3 n=9, SCA6 n=9) and 15 healthy volunteers were studied. Voxel-based morphometry (VBM) was applied to segmented grey matter (GM) and white matter (WM) of high-resolution T1-weighted brain volumes of each group. We found reduction of grey matter in the pons as well as in the vermis in SCA3 as compared to control subjects. In SCA6 significant grey matter loss was found in hemispheric lobules bilaterally as well as in the vermis. White matter analysis revealed significant changes in SCA3, especially in the pons, in the white matter surrounding the dentate nucleus (DN) and in the cerebellar peduncles, whereas no significant white matter reduction was found in SCA6 patients. Our results demonstrate different patterns of grey and white matter affection detected by magnetic resonance imaging (MRI) in SCA3 and SCA6 patients, confirming the pathological concept of cortical cerebellar atrophy in SCA6. In contrast, SCA3 represents a form of ponto-cerebellar atrophy with predominant affection of pontine nuclei and fibre tracts.

背景与目标： ：本研究的目的是检查3型和6型脊髓小脑共济失调（SCA）的小脑和/或脑干萎缩的不同模式。研究了18例患者（SCA3 n = 9，SCA6 n = 9）和15名健康志愿者。将基于体素的形态计量学（VBM）应用于每组高分辨率T1加权脑体积的分段灰质（GM）和白质（WM）。我们发现与对照组相比，SCA3的脑桥和s中的灰质减少。在SCA6中，在双侧的半球小叶以及在mis骨中都发现了明显的灰质损失。白质分析显示，SCA3的显着变化，尤其是在脑桥，齿状核（DN）周围的白质和小脑梗的脑桥中，特别是在脑桥中，而在SCA6患者中未发现显着的白质减少。我们的结果表明，在SCA3和SCA6患者中通过磁共振成像（MRI）检测到不同的灰白色和白色物质影响模式，证实了SCA6皮质小脑萎缩的病理学概念。相反，SCA3代表了一种桥脑小脑萎缩的形式，主要影响桥脑核和纤维束。

BACKGROUND & AIMS: Interleukin-1beta (IL-1beta) is closely involved in liver disorders. IL-1beta produces nitric oxide (NO) in vascular smooth muscle cells and relaxes vascular smooth muscle via cyclic guanosine 3',5'-monophosphate (cGMP). In this study, we evaluated the relaxing effect of IL-1beta on cultured Ito cells. Ito cells were isolated from the livers of male Wistar rats and cultured for 24 hours. Immunolocalization of inducible nitric oxide synthase (iNOS) and cGMP and intensity of fluorescence of cGMP were examined using a confocal laser microscope. Ito cells were treated with 0, 200, and 1,000 pmol/L IL-1beta, and the intracellular cGMP concentration was measured after 12 hours. Moreover, Ito cells treated with 200 and 1,000 pmol/L IL-1beta and not treated with IL-1beta were observed over 12 hours, and the area of the same Ito cell was compared before and after the addition of IL-1beta. Next, effects of N(G)-monomethyl-L-arginine (L-NMMA) and S-nitroso-N-acetyl-DL-penicillamine (SNAP) on Ito cell relaxation by IL-1beta treatment were examined. In Ito cells, immunofluorescence of iNOS was observed, and fluorescent intensity of cGMP increased after addition of IL-1beta. Intracellular cGMP concentration increased dose-dependently after addition of IL-1beta. Cell area significantly increased in the IL-1beta-treated group compared with the untreated group. Relaxation of Ito cells by IL-1beta treatment was inhibited by L-NMMA in a dose-dependent manner, but was enhanced by SNAP. These results indicate that IL-1beta produces NO in cultured Ito cells and relaxes the cells via cGMP.

背景与目标： 白介素-1β（IL-1beta）与肝脏疾病密切相关。 IL-1beta通过环状鸟苷3'，5'-单磷酸（cGMP）在血管平滑肌细胞中产生一氧化氮（NO），并使血管平滑肌松弛。在这项研究中，我们评估了IL-1β对培养的Ito细胞的松弛作用。从雄性Wistar大鼠的肝脏中分离出Ito细胞，并培养24小时。使用共聚焦激光显微镜检查诱导型一氧化氮合酶（iNOS）和cGMP的免疫定位和cGMP的荧光强度。用0、200和1,000 pmol / L IL-1beta处理Ito细胞，并在12小时后测量细胞内cGMP浓度。此外，在12小时内观察到用200和1,000 pmol / L IL-1beta处理且未用IL-1beta处理的Ito细胞，并且在添加IL-1beta之前和之后比较了同一个Ito细胞的面积。接下来，研究了N（G）-单甲基-L-精氨酸（L-NMMA）和S-亚硝基-N-乙酰基-DL-青霉胺（SNAP）对通过IL-1beta处理的Ito细胞松弛的影响。在Ito细胞中，观察到iNOS的免疫荧光，添加IL-1β后cGMP的荧光强度增加。加入IL-1beta后，细胞内cGMP浓度呈剂量依赖性增加。与未治疗组相比，IL-1β治疗组的细胞面积显着增加。 L-1NMMA以剂量依赖的方式抑制了通过IL-1beta处理的Ito细胞的松弛，但被SNAP增强了。这些结果表明IL-1β在培养的Ito细胞中产生NO，并通过cGMP使细胞松弛。

BACKGROUND & AIMS: :The signaling pathways mediating lysophosphatidic acid (LPA)-stimulated PKD(2) activation and the potential contribution of PKD(2) in regulating LPA-induced interleukin 8 (IL-8) secretion in nontransformed, human colonic epithelial NCM460 cells were examined. Treatment of serum-deprived NCM460 cells with LPA led to a rapid and striking activation of PKD(2), as measured by in vitro kinase assay and phosphorylation at the activation loop (Ser706/710) and autophosphorylation site (Ser876). PKD(2) activation induced by LPA was abrogated by preincubation with selective PKC inhibitors GF-I and Ro-31-8220 in a dose-dependent manner. These inhibitors did not have any direct inhibitory effect on PKD(2) activity. LPA induced a striking increase in IL-8 production and stimulated NF-kappaB activation, as measured by NF-kappaB-DNA binding, NF-kappaB-driven luciferase reporter activity, and IkappaBalpha phosphorylation. PKD(2) gene silencing utilizing small interfering RNAs targeting distinct PKD(2) sequences dramatically reduced LPA-stimulated NF-kappaB promoter activity and IL-8 production. PKD(2) activation is a novel early event in the biological action of LPA and mediates LPA-stimulated IL-8 secretion in NCM460 cells through a NF-kappaB-dependent pathway. Our results demonstrate, for the first time, the involvement of a member of the PKD family in the production of IL-8, a potent proinflammatory chemokine, by epithelial cells.

背景与目标： ：审查了介导溶血磷脂酸（LPA）刺激的PKD（2）激活和PKD（2）在调节LPA诱导的非转化型人结肠上皮NCM460细胞中LPA诱导的白介素8（IL-8）分泌中的潜在作用。用体外LPA处理血清缺乏的NCM460细胞会导致PKD（2）迅速而惊人的活化，这是通过体外激酶测定和活化环（Ser706 / 710）和自磷酸化位点（Ser876）的磷酸化来测量的。通过与选择性PKC抑制剂GF-1和Ro-31-8220呈剂量依赖性方式进行预孵育，可以消除LPA诱导的PKD（2）激活。这些抑制剂对PKD（2）活性没有任何直接的抑制作用。如通过NF-κB-DNA结合，NF-κB驱动的萤光素酶报道分子活性和IkappaBalpha磷酸化所测量，LPA诱导IL-8产量显着增加并刺激NF-κB活化。 PKD（2）基因沉默利用针对不同的PKD（2）序列的小干扰RNA，大大降低了LPA刺激的NF-κB启动子活性和IL-8的产生。 PKD（2）激活是LPA的生物学作用中的一个新的早期事件，并通过NF-κB依赖性途径介导NCM460细胞中LPA刺激的IL-8分泌。我们的结果首次证明，上皮细胞参与了PKD家族成员参与IL-8（一种有效的促炎趋化因子）的生产。

BACKGROUND & AIMS: PURPOSE:To assess the technical feasibility, thrombogenicity, and biocompatibility of a new biodegradable poly-L-lactic acid (PLLA) anastomotic stent. METHODS:A polytetrafluoroethylene bifurcated graft was implanted in 17 pigs through a midline abdominal incision. After transverse graft incision, 17 316L stainless steel stents and 17 PLLA stents were randomly implanted at both iliac anastomotic sites and deployed with a 6-mm balloon under direct vision without angiography. Intended follow-up was 1 week in 6 pigs receiving oral acetylsalicylic acid (ASA) and in 7 pigs receiving ASA/clopidogrel; 4 pigs receiving ASA/clopidogrel were followed for 6 weeks. At the end of the study, the segments containing the stents were surgically explanted and processed for histology to measure the mean luminal diameter, intimal thickness, and the vascular injury and inflammation scores. RESULTS:Initial technical success of stent placement was achieved in all animals without rupture of the suture. Two pigs died (unrelated to the stent) at 3 days after operation (1 in groups A and B). At 1 week, all PLLA stents showed thrombotic occlusion with the use of ASA alone. In contrast, all PLLA stents remained patent with concurrent administration of ASA/clopidogrel. All metal stents were patent regardless of the antiplatelet regimen. The mean luminal diameter of patent PLLA stents (4.13+/-0.17 mm) was comparable to metal stents (4.27+/-0.35 mm, p=0.78) at 1 week, but significantly diminished at 6 weeks (3.21+/-0.44 versus 4.19+/-0.18 mm, p=0.005). Histological analysis showed no signs of excessive recoil. PLLA stents induced a higher inflammation score (1.79+/-0.56) and more intimal hyperplasia (0.34+/-0.11 mm) compared to metal stents [1.27+/-0.44 mm (p<0.001) and 0.18+/-0.04 mm (p=0.006), respectively] at 6 weeks. Vascular injury was comparable between PLLA and metal stents. CONCLUSION:Biodegradable PLLA stents showed higher thrombogenicity and reduced patency compared to metal stents during early follow-up. Although ASA and clopidogrel prevented thrombotic occlusion, the increased inflammatory response and neointima formation remain major concerns of PLLA stents. A solution to this problem might be the incorporation of anti-inflammatory drugs into the PLLA stent.

背景与目标： 目的：评估新型可生物降解的聚-L-乳酸（PLLA）吻合支架的技术可行性，血栓形成性和生物相容性。
方法：通过腹部中线切口将聚四氟乙烯分叉移植物植入17只猪。横向移植后，将17个316L不锈钢支架和17个PLLA支架随机植入两个both吻合部位，并在不进行血管造影的情况下在直视下用6毫米球囊展开。预期的随访结果是：6头接受口服乙酰水杨酸（ASA）的猪和7头接受ASA /氯吡格雷的猪。对4只接受ASA /氯吡格雷的猪进行了6周的随访。在研究结束时，将包含支架的节段进行外科手术切除并进行组织学处理，以测量平均腔直径，内膜厚度以及血管损伤和炎症评分。
结果：在所有动物中均未发生缝线破裂的情况下，支架植入取得了初步的技术成功。手术后3天有2头猪死亡（与支架无关）（A和B组为1只）。在第1周，仅使用ASA时，所有PLLA支架均显示血栓闭塞。相反，所有PLLA支架在同时使用ASA /氯吡格雷的情况下仍保持专利。无论抗血小板方案如何，所有金属支架均已获得专利。专利PLLA支架的平均腔直径（4.13 /-0.17 mm）在1周时可与金属支架（4.27 /-0.35 mm，p = 0.78）相媲美，但在6周时显着减小（3.21 // 0.44对4.19 /-） 0.18毫米，p = 0.005）。组织学分析显示没有过度后坐的迹象。与金属支架[1.27 /-0.44 mm（p <0.001）和0.18 /-0.04 mm（p = 0.006）相比，PLLA支架引起更高的炎症评分（1.79 /-0.56）和更多的内膜增生（0.34 /-0.11 mm）。分别]在6周。 PLLA和金属支架之间的血管损伤相当。
结论：在早期随访中，与金属支架相比，可生物降解的PLLA支架显示出更高的血栓形成性和通畅性降低。尽管ASA和氯吡格雷预防了血栓闭塞，但炎症反应和新内膜形成的增加仍然是PLLA支架的主要关注点。解决该问题的方法可能是将抗炎药掺入PLLA支架中。

BACKGROUND & AIMS: :The gram-negative bacterium Bartonella henselae is capable of causing angiogenic lesions as a result of infection. Previously, it has been shown that B. henselae infection can result in production of the chemokine interleukin-8 (IL-8). In this study, we demonstrated that monocytes, endothelial cells, and hepatocytes produce IL-8 in response to B. henselae infection. We also investigated the role of IL-8 in B. henselae-induced endothelial cell proliferation and capillary tube formation. Both in vitro angiogenesis assays were IL-8 dependent. B. henselae-mediated inhibition of apoptosis, as indicated by gene expression of Bax and Bcl-2, was also shown to be IL-8 dependent in endothelial cells. Furthermore, infection of endothelial cells with B. henselae stimulated upregulation of the IL-8 chemokine receptor CXCR2. Infection of human endothelial cells by B. henselae resulting in IL-8 production likely plays a central role in the ability of this organism to cause angiogenesis during infection.

背景与目标： 革兰氏阴性细菌汉氏巴尔通体（Bartonella henselae）能够由于感染而引起血管生成性病变。以前，已经证明，亨氏芽孢杆菌感染可以导致趋化因子白介素8（IL-8）的产生。在这项研究中，我们证明了单核细胞，内皮细胞和肝细胞对B. henselae感染产生IL-8。我们还研究了IL-8在B. henselae诱导的内皮细胞增殖和毛细管形成中的作用。两种体外血管生成测定都是IL-8依赖性的。如Bax和Bcl-2的基因表达所表明的，B。henselae介导的凋亡抑制在内皮细胞中也显示为IL-8依赖性。此外，用汉逊芽孢杆菌感染内皮细胞刺激了IL-8趋化因子受体CXCR2的上调。亨氏芽孢杆菌对人内皮细胞的感染导致IL-8的产生可能在该生物体在感染过程中引起血管生成的能力中起着核心作用。

BACKGROUND & AIMS: :We have constructed a physical map of a > 2-Mb region on mouse chromosome 6 that contains the natural killer gene complex (NKC). The map comprises a contig of 14 overlapping yeast artificial chromosomes onto which we positioned 25 NKC markers. NKC genetically linked genes encode > 17 proteins that directly control innate NK cell-mediated tumor lysis and disease resistance. Herein we show that Nkrp1 genes are clustered in a region flanked by A2m and Cd69 genes and that most Ly49 genes are clustered in a distal region -1 Mb distant. Importantly, syntenic intervals of mouse chromosome 6 and human chromosome 12p that include the NKC are conserved. NKC species conservation suggests that the human NKC may contain orthologues for the mouse viral disease resistance genes, Cmv1 and Rmp1. The high-resolution NKC map will facilitate investigation of NKC gene regulation and identification of phenotypically defined gene products that confer NK cell defense against viral pathogens.

背景与目标： ：我们在小鼠染色体6上构建了一个大于2-Mb区域的物理图，其中包含自然杀伤基因复合体（NKC）。该图包括14个重叠的酵母人工染色体的重叠群，我们在其上定位了25个NKC标记。 NKC遗传连锁基因编码> 17种蛋白质，这些蛋白质直接控制先天NK细胞介导的肿瘤溶解和疾病抵抗力。在本文中，我们显示Nkrp1基因聚集在A2m和Cd69基因侧翼的区域中，而大多数Ly49基因聚集在距离-1 Mb远的区域中。重要的是，保留了包含NKC的小鼠6号染色体和人类12p号染色体的同音间隔。 NKC物种保守性表明，人NKC可能含有小鼠病毒疾病抗性基因Cmv1和Rmp1的直向同源物。高分辨率的NKC图谱将有助于NKC基因调控的研究和表型定义的基因产物的鉴定，这些产物赋予NK细胞防御病毒病原体的能力。

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BH4的合成，从而揭示了所有细菌PTPS同源物。

BACKGROUND & AIMS: A primer extension (toeprinting) assay was used to monitor selection by ribosomes of the first versus the second AUG codon as a function of introducing mutations on the 3' side (positions +4, +5 and +6) of the first AUG codon. Six different flanking codons starting with G (GCG, GCU, GCC, GCA, GAU and GGA) strongly augmented selection of AUG#1 when compared with matched mRNAs that had A or C instead of G in position +4. Augmentation by G in position +4 failed only when it was combined with U in position +5, as in the sequence augGUA. In contrast with the usual enhancing effect of introducing G in position +4, most mutations in position +5 had no discernible effect, as shown with the series augANA (where N = C, A, G or U) and the series augCNA. AUG codon recognition was also unaffected by mutations in position +6, as shown by testing four mRNAs that had augCCN as the start site. Thus the primary sequence context that augments the recognition of AUG start codons does not appear generally to extend beyond G in position +4. When the toeprinting assay was used with mRNAs that initiate translation at CUG instead of AUG, cugGAU was not recognized better than cugGGU, contradicting the hypothesis that initiation at non-AUG codons might be favored by A instead of G in position +5.

背景与目标： 使用引物延伸（印迹）测定法来监测第一和第二AUG密码子的核糖体的选择，其作为在第一AUG密码子的3'侧（位置4、5和6）上引入突变的函数。与匹配的在位置4有A或C而不是G的mRNA相比，以G开头的六个不同的侧翼密码子（GCG，GCU，GCC，GCA，GAU和GGA）大大增强了对AUG＃1的选择。仅当它与位置5的U组合时失败，如序列augGUA所示。与在位置4引入G的通常增强作用相反，在位置5的大多数突变没有明显的作用，如augANA系列（其中N = C，A，G或U）和augCNA系列所示。 AUG密码子识别也不受位置6突变的影响，如测试以augCCN为起始位点的四个mRNA所显示的。因此，增强对AUG起始密码子识别的一级序列通常不会延伸到位置4的G范围之外。当将脚印法与在CUG而非AUG处起始翻译的mRNA一起使用时，cugGAU的识别度不如cugGGU，与以下假设相矛盾：在非AUG密码子处的起始可能会被位置5的A而不是G所偏爱。

BACKGROUND & AIMS: Interleukin-1 (IL-1) has been implicated in chronic and acute cerebral neuropathologies. IL-1 receptor antagonist (IL-1ra), a naturally occurring protein that binds to IL-1 receptors without inducing signal transduction, blocks several actions of IL-1. IL-1ra acts at the local level and it also circulates in the bloodstream. We now report evidence for a biological function of IL-1ra in the brain as an endogenous neuroprotective molecule. Cerebral expression of IL-1ra mRNA is induced rapidly by focal cerebral ischemia in rats, and inhibition of the action of IL-1ra, by passive immuno-neutralization, markedly enhances ischemic damage. To our knowledge this is the first report of an action of endogenous IL-1ra in the brain. Control of IL-1ra expression or action may therefore provide a useful therapeutic strategy to limit acute neurodegeneration.

背景与目标： 白介素-1（IL-1）已牵涉到慢性和急性脑神经病理学。 IL-1受体拮抗剂（IL-1ra）是一种与IL-1受体结合而不诱导信号转导的天然蛋白质，可阻断IL-1的多种作用。 IL-1ra在局部起作用，并且也在血液中循环。我们现在报告IL-1ra在脑中作为内源性神经保护分子的生物学功能的证据。大鼠局灶性脑缺血可快速诱导IL-1ra mRNA的脑表达，并且被动免疫中和抑制IL-1ra的作用可显着增强缺血性损伤。据我们所知，这是大脑中内源性IL-1ra作用的首次报道。因此，控制IL-1ra的表达或作用可能为限制急性神经变性提供了有用的治疗策略。

BACKGROUND & AIMS: :Given the paucity of data on the distribution of serotonin (5-HT) receptors of type 6 (5-HT(6)) in the human brain, the aim of this study was to investigate their distribution in postmortem human prefrontal cortex, striatum and hippocampus by either immunohistochemical or immunofluorescence techniques. The brain samples were obtained from 6 subjects who had died for causes not involving primarily or secondarily the CNS. The 5-HT(6) receptor distribution was explored by the [(125)I]SB-258585 binding to brain membranes followed by immunohistochemical and immunofluorescence evaluations. A specific [(125)I]SB-258585 binding was detected in all the regions under investigation, whilst the content in the hippocampus and cortex being about 10-30 times lower than in the striatum. Immunohistochemistry and double-label immunofluorescence microscopy experiments, carried out in the prefrontal cortex and hippocampus only, since data in the striatum were already published, showed the presence of 5-HT(6) receptors in both pyramidal and glial cells of prefrontal cortex, while positive cells were mainly pyramidal neurons in the hippocampus. The heterogeneous distribution of 5-HT(6) receptors provides a preliminary explanation of how they might regulate different functions in different brain areas, such as, perhaps, brain trophism in the cortex and neuronal firing in the hippocampus. This study, taking into account all the limitations due to the postmortem model used, represents the starting point to explore the 5-HT(6) receptor functionality and its sub-cellular distribution.

背景与目标： ：鉴于缺乏6型血清素（5-HT）受体（5-HT（6））在人脑中的分布的数据，本研究的目的是研究它们在死后人类前额叶皮层，纹状体中的分布免疫组织化学或免疫荧光技术检测海马和海马。脑样本是从6名因主要或次要不涉及CNS的原因死亡的受试者中获得的。通过[（125）I] SB-258585与脑膜的结合，然后进行免疫组织化学和免疫荧光评估，探索了5-HT（6）受体的分布。在所有研究区域中均检测到特定的[（125）I] SB-258585结合，而海马和皮质中的含量比纹状体低约10-30倍。仅在前额叶皮层和海马体中进行的免疫组织化学和双标记免疫荧光显微镜实验，因为纹状体中的数据已经发表，显示在前额叶皮层的锥体细胞和神经胶质细胞中都存在5-HT（6）受体，而阳性细胞主要是海马中的锥体神经元。 5-HT（6）受体的异质分布为它们如何在不同的大脑区域调节不同的功能提供了初步的解释，例如，大脑皮层的营养和海马神经元的放电。这项研究，考虑到由于使用死后模型的所有限制，代表了探索5-HT（6）受体功能及其亚细胞分布的起点。

BACKGROUND & AIMS: :Abstract Type III interferon (IFN-λ) is a novel member of the interferon family, which preferentially promotes antiviral responses from epithelial cells and cooperates with type I IFNs in the clearance of viral infections. However, the effect of mIFN-λ2 to the LA795 lung adenocarcinoma cell is largely unknown. In this study, we transfected Ad-mIFN-λ2 vector into LA795 tumor-bearing mice to explore the effect of mIFN-λ2 on the proliferation of LA795 lung adenocarcinoma cell and on the immune response of the mice. Transfected by Ad-mIFN-λ2 vector, a significant decrease in the tumor growth, the subcutaneous tumor necrosis, cystic degeneration, and tumor apoptosis were more evident; at the same time, mIFN-λ2 protein and gene were significantly more expressed. And, flow cytometry analysis suggested that CD3(+)CD4(+), CD3(+)CD8(+), and NK (CD3(-)CD49(+)) cells were all significantly increased after transfected by Ad-mIFN-λ2. The study demonstrated that recombinant Ad-mIFN-λ2 transfection effectively inhibited the growth of LA795 lung adenocarcinoma cell, which may work through inducing apoptosis of tumor cell and regulating cell immune response.

背景与目标： 摘要：III型干扰素（IFN-λ）是干扰素家族的一个新成员，它优先促进上皮细胞的抗病毒反应，并与I型干扰素协同作用以清除病毒感染。然而，很大程度上未知mIFN-λ2对LA795肺腺癌细胞的作用。在这项研究中，我们将Ad-mIFN-λ2载体转染到LA795荷瘤小鼠中，以研究mIFN-λ2对LA795肺腺癌细胞增殖和小鼠免疫反应的影响。 Ad-mIFN-λ2载体转染后，肿瘤生长明显减少，皮下肿瘤坏死，囊性变性和肿瘤细胞凋亡更为明显。同时，mIFN-λ2蛋白和基因的表达明显增加。并且，流式细胞仪分析表明，用Ad-mIFN-λ2转染后，CD3（）CD4（），CD3（）CD8（）和NK（CD3（-）CD49（））细胞均显着增加。研究表明重组Ad-mIFN-λ2转染有效抑制了LA795肺腺癌细胞的生长，其作用可能是通过诱导肿瘤细胞凋亡和调节细胞免疫应答来实现的。

BACKGROUND & AIMS: :Pneumocystis pneumonia (PCP) incidence was decreased in renal transplant thanks to prophylaxis, recommended during the first months after transplantation. However, many late PCP cases are observed after the first 6 months and recommendations to maintain or reintroduce prophylaxis are lacking. The objective of the study was to identify risk factors to guide the individual prescription of prophylaxis, 6 months after transplantation. Thirty-three late PCP cases were identified between 1995 and 2012 in Lille Hospital, France, and were compared to 72 randomized controls transplant recipients. In univariate analysis, age of donor (>48 years), retransplantation, a decrease glomerular filtration rate (≤45 mL/min), induction therapy mediated by anti-thymocyte globulin (ATG), steroid maintenance, high calcineurin inhibitors (CNI) doses (tacrolimus ≥0.5 mg/kg/day and cyclosporine ≥2.1 mg/kg/day), and cytomegalovirus (CMV) infection were significantly associated with PCP. In multivariate analysis, ATG (hazard ratio [HR]: 2.4 [1.1-5.4]), steroid therapy (HR: 3.1 [1.20-7.84], CNI (HR: 2.9 [1.28-6.38], and CMV (HR: 6.1 [2.74-16.33] remained associated with late PCP. In conclusion, we confirm that intensive immunosuppressive regimen and CMV infection are critical risk factors for late PCP and should be taken into account to decide on maintenance or reintroduction of a prophylactic treatment.

背景与目标： ：由于预防，建议在移植后的头几个月内，减少肾脏移植中肺囊虫性肺炎（PCP）的发生率。但是，在头6个月后观察到许多晚期PCP病例，缺乏维持或重新引入预防措施的建议。该研究的目的是确定危险因素，以指导移植后6个月进行预防的个体处方。在1995年至2012年之间，法国里尔医院确定了33例晚期PCP病例，并将其与72名随机对照移植受者进行了比较。在单因素分析中，供体年龄（> 48岁），再移植，肾小球滤过率降低（≤45mL / min），抗胸腺细胞球蛋白（ATG）介导的诱导治疗，类固醇维持，钙调神经磷酸酶抑制剂（CNI）剂量高他克莫司≥0.5mg / kg /天，环孢菌素≥2.1mg / kg /天）和巨细胞病毒（CMV）感染与PCP显着相关。在多变量分析中，ATG（危险比[HR]：2.4 [1.1-5.4]），类固醇治疗（HR：3.1 [1.20-7.84]，CNI（HR：2.9 [1.28-6.38]）和CMV（HR：6.1 [ [2.74-16.33]仍与晚期PCP相关，总的来说，我们确认强化免疫抑制方案和CMV感染是晚期PCP的关键危险因素，应考虑维持或重新引入预防性治疗。