BACKGROUND & AIMS: :Systemic lupus erythematosus (SLE) is a chronic systemic inflammatory disease. Autoantibodies (autoAbs) against double-stranded DNA (ds DNA), the hallmark of lupus, are produced and maintained by the interaction between auto-reactive B cells and CD4+ T cells. This interplay is controlled by the CD28/CD80-86/CTLA-4 axis. Here we investigated whether selective blockade of CD28-CD80/86 co-stimulatory interactions abrogates lupus nephritis development in a murine model of SLE. To this aim, NZB/NZW F1 mice were treated for 3 months, either with an anti-CD28 Fab' fragment or a control Fab'-IgG. The effect of CD28 blockade on lupus nephritis onset, survival, production of anti-ds DNA antibodies and costimulatory molecules was evaluated. CD28 blockade prevented the development of lupus nephritis and prolonged survival during the 3-month treatment and 12 weeks after. Furthermore, the production of anti-ds DNA autoAbs was decreased. Lastly, the protective effect of CD28 blockade was associated with increased intrarenal expression of the immunoregulatory molecule, Indoleamine 2, 3-dioxygenase, of the co-inhibitory receptor programmed cell-Death - 1 (PD-1) and of its ligand programmed death ligand - 1 (PDL-1).In conclusion, CD28 blockade prevented the development of lupus nephritis in NZB/NZW F1 mice. This immunomodulatory strategy is a promising candidate for SLE therapy in humans.

背景与目标： 系统性红斑狼疮（SLE）是一种慢性全身性炎症性疾病。通过自身反应性B细胞和CD4 T细胞之间的相互作用，产生并维持针对狼疮的标志性双链DNA（ds DNA）的自身抗体（autoAbs）。这种相互作用是由CD28 / CD80-86 / CTLA-4轴控制的。在这里，我们研究了CD28-CD80 / 86共刺激相互作用的选择性阻断是否消除了狼疮性肾炎在SLE鼠模型中的发展。为了这个目的，将NZB / NZW F1小鼠用抗CD28 Fab'片段或对照Fab'-IgG治疗3个月。评估了CD28阻断对狼疮肾炎发作，生存，抗ds DNA抗体产生和共刺激分子的影响。 CD28阻滞剂在3个月的治疗期间和12周后预防了狼疮性肾炎的发展并延长了生存期。此外，抗ds DNA autoAb的产生减少。最后，CD28阻断的保护作用与免疫调节分子吲哚胺2、3-二加氧酶，共抑制受体编程的细胞死亡-1（PD-1）及其配体编程的死亡配体的肾内表达增加有关。 -1（PDL-1）。总之，CD28阻断可预防NZB / NZW F1小鼠的狼疮性肾炎。这种免疫调节策略是人类SLE治疗的有希望的候选者。

BACKGROUND & AIMS: :A novel series of potent and efficacious factor Xa inhibitors which possesses sulfoximine moiety as novel S4 binding element in anthranilamide chemotype has been identified. Lead optimization at this novel P4 group led to many potent factor Xa inhibitors with excellent anticoagulant activity in human plasma. Selected compounds were dosed orally in rats and checked for their ex vivo prothrombin time prolonging activity, which resulted in identification of compound 5-chloro-N-(5-chloropyridin-2-yl)-2-(4-(N-(2-(diethylamino)acetyl)-S-methylsulfonimidoyl)benzamido)benzamide (18f). The detailed pharmacokinetic evaluation and subsequent metabolism study of 18f suggested the presence of an active metabolite. The compound 18f and its active metabolite 18b demonstrated excellent in vivo efficacy in both arterial and venous thrombosis model in rats and were found to be highly selective against related serine proteases. Based on this promising profile, compound 18f was selected for further evaluation.

背景与目标： ：已经鉴定了一系列有效且有效的因子Xa抑制剂，其具有硫肟亚胺部分作为邻氨基苯甲酰胺化学型中的新的S4结合元件。在这个新颖的P4组中的前导优化导致了许多有效的Xa抑制剂在人血浆中具有出色的抗凝活性。在大鼠中口服选择的化合物并检查其离体凝血酶原时间延长活性，从而鉴定出化合物5-氯-N-（5-氯吡啶-2-基）-2-（4-（N-（2 -（二乙氨基）乙酰基）-S-甲基磺酰亚胺基）苯甲酰胺基）苯甲酰胺（18f）。 18f的详细药代动力学评估和随后的代谢研究表明存在活性代谢物。化合物18f及其活性代谢物18b在大鼠的动脉和静脉血栓形成模型中均显示出优异的体内功效，并且被发现对相关的丝氨酸蛋白酶具有高度选择性。基于这一有前途的概况，选择了化合物18f进行进一步评估。

BACKGROUND & AIMS: :Fluoxetine (FLX) the active pharmaceutical ingredient (API) in Prozac(®) is a widely prescribed psychoactive drug which ubiquitous occurrence in the aquatic environment is associated to a poor removal rate in waste-water treatment plant (WWTP) systems. This API acts as a selective serotonin reuptake inhibitor (SSRI) frequently reported to cause disrupting effects in non-target species. The objective of this study includes a multibiomarker response evaluation on mussel Mytilus galloprovincialis during two weeks exposure to 75 ng L(-1) FLX assessing antioxidant enzymes activities--superoxide dismutase (SOD), catalase (CAT) and glutathione-S-transferase (GST); lipid peroxidation (LPO), acetylcholinesterase (AChE) neurotoxic response and endocrine disruption through alkali-labile phosphates (ALP) indirect measurement of vitellogenin-like proteins. Results show transient tissue-specific enzymatic responses and damage affecting mostly mussel gills. However, the clear ALP levels inhibition throughout time in both sex-differentiated gonads gives evidence to FLX reinforced action as an endocrine disruptor rather than an oxidative or neurotoxic inducer.

背景与目标： ：Prozac®中的活性药物成分（API）氟西汀（FLX）是一种广泛使用的精神活性药物，在水生环境中普遍存在，与废水处理厂（WWTP）系统中的去除率差有关。此API作为一种选择性血清素再摄取抑制剂（SSRI）经常报道引起非目标物种干扰作用。这项研究的目的包括在暴露于75 ng L（-1）FLX的两周期间对贻贝贻贝贻贝进行多生物标志物反应评估，以评估抗氧化酶活性-超氧化物歧化酶（SOD），过氧化氢酶（CAT）和谷胱甘肽S-转移酶（消费税）;脂质过氧化（LPO），乙酰胆碱酯酶（AChE）的神经毒性反应和通过碱不稳定的磷酸盐（ALP）间接测量卵黄蛋白原样蛋白的内分泌干扰。结果表明，短暂的组织特异性酶促反应和损害主要影响贻贝g。然而，在两个性别分化的性腺中，随着时间的推移，明显的ALP水平抑制作用为FLX作为内分泌干扰物而不是氧化或神经毒性诱导物的增强作用提供了证据。

BACKGROUND & AIMS: :The present study was designed to investigate whether taxol in combination with cyclooxygenase (COX) inhibitors could be superior on inhibitory effect of ovarian cancer growth than taxol alone as drug therapy of mice implanted with human ovarian carcinoma cell line SKOV-3. The animals were treated with 100 mg/ kg celecoxib (a COX-2 selective inhibitor) alone or in combination with 3 mg/kg SC-560 (a COX-1 selective inhibitor) by gavage twice a day, 20mg/kg taxol alone by intraperitoneal (IP) once a week or in combination with celecoxib, or SC-560/celecoxib/taxol for 3 weeks. To test the mechanism of the combination treatment, the index of cell proliferation, expression of cyclin D1, and microvessel density (MVD) in tumor tissues were determined by immunohistochemistry and the index of apoptotic cells by the terminal-deoxynucleotidyl-transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) method. Mean tumor volume in the SC-560/celecoxib/taxol group was first significantly lower than control at day 14 (p < 0.05). In the SC-560/celecoxib/taxol group, the index of cell proliferation and apoptosis and quantification of cyclin D1-postive cells were 6.93%, 69.62%, and 19.14%, respectively, which are statistically significant compared with those of the control group (29.85%, p < 0.001; 32.81% and 36.99%, both p < 0.05). Statistical significance on MVD was observed between the SC-560/celecoxib/taxol (39.57 +/- 4.98) and the control (73.2 +/- 1.96) group (p < 0.001). Our results suggest that the combined antitumor efficacy of taxol and COX inhibitors may be superior to taxol alone as drug therapy against ovarian cancer in mice, and that synergism of the combination treatment in part may be mediated through accelerated apoptosis and suppression of cell proliferation and angiogenesis.

背景与目标： ：本研究旨在研究紫杉醇与环加氧酶（COX）抑制剂联合使用对人卵巢癌细胞株SKOV-3植入小鼠的卵巢癌生长抑制作用是否比单独使用紫杉醇更好。每天两次分别用100 mg / kg celecoxib（一种COX-2选择性抑制剂）或与3 mg / kg SC-560（一种COX-1选择性抑制剂）联合饲喂动物，分别用20 mg / kg紫杉醇治疗。每周一次腹膜内（IP）或与celecoxib或SC-560 / celecoxib /紫杉醇联合使用3周。为了测试联合治疗的机制，通过免疫组织化学测定肿瘤组织中细胞增殖指数，细胞周期蛋白D1的表达和微血管密度（MVD），并用末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸来检测凋亡细胞的指数。缺口标记（TUNEL）方法。第14天时，SC-560 /塞来昔布/紫杉醇组的平均肿瘤体积首先显着低于对照组（p <0.05）。在SC-560 /塞来昔布/紫杉醇组中，细胞周期cyclin D1阳性细胞的增殖指数和凋亡指数分别为6.93％，69.62％和19.14％，与对照组相比有统计学意义。 （29.85％，p ＜0.001； 32.81％和36.99％，均p ＜0.05）。在SC-560 / celecoxib /紫杉醇（39.57 /-4.98）和对照组（73.2 /-1.96）组之间观察到MVD的统计显着性（p <0.001）。我们的研究结果表明，紫杉醇和COX抑制剂的联合抗肿瘤药效可能优于单独使用紫杉醇作为抗小鼠卵巢癌的药物疗法，并且联合治疗的协同作用可能部分通过加速凋亡以及抑制细胞增殖和血管生成来介导。 。

BACKGROUND & AIMS: :The phototransformation pathways of protochlorophyllide forms were studied in 8-14-day-old leaves of dark-germinated wheat (Triticum aestivum L.) using white, 632.8 nm He-Ne laser and 654 nm laser diode light. The photon flux density (PFD) values (0.75-360 μmol photons m(-2) s(-1)), the illumination periods (20 ms-10 s) and the temperature of the leaves (between -60 °C and room temperature) were varied. The 77 K fluorescence spectra of partially phototransformed leaves showed gradual accumulation or even the dominance of the 675 nm emitting chlorophyllide or chlorophyll form at room temperature with 632.8 nm of PFD less than 200 μmol photons m(-2) s(-1) or with 654 nm of low PFD (7.5 μmol photons m(-2) s(-1)) up to 1 s. Longer wavelength (685 or 690 nm) emitting chlorophyllide forms appeared at illuminations under -25 °C with both laser lights or at room temperature when the PFD values were higher or the illumination period was longer than above. We concluded that the formation of the 675 nm emitting chlorophyllide form does not indicate the direct photoactivity of the 633 nm emitting protochlorophyllide form; it can derive from 644 and 657 nm forms via instantaneous disaggregation of the newly-produced chlorophyllide complexes. The disaggregation is strongly influenced by the molecular environment and the localization of the complex.

背景与目标： ：在白色发芽的小麦（Triticum aestivum L.）的8-14天大的叶片中，使用白色，632.8 nm He-Ne激光和654 nm激光二极管光研究了原叶绿素内酯形式的光转化途径。光子通量密度（PFD）值（0.75-360μmol光子m（-2）s（-1）），光照时间（20 ms-10 s）和叶片温度（-60°C和室温之间）温度）各不相同。部分光转化的叶片的77 K荧光光谱在室温下，632.8 nm的PFD小于200μmol光子m（-2）s（-1）或与654 nm的低PFD（7.5μmol光子m（-2）s（-1））长达1 s。当PFD值较高或照射时间长于-25°C时，在两种激光下或在室温下照射时，会发出较长波长（685或690 nm）的叶绿素形式。我们得出的结论是，发出675 nm的叶绿素的形式的形成并不表明发出633 nm的原叶绿素的形式的直接光敏性。它可以通过瞬时分解新产生的叶绿素配合物而从644和657 nm的形式中获得。分解受到分子环境和配合物局部的强烈影响。

BACKGROUND & AIMS: 1. Selective 5-hydroxytryptamine (5-HT; serotonin) reuptake inhibitors (SSRIs) cause a greater increase in extracellular 5-HT in the forebrain when the somatodendritic 5-HT1A autoreceptor is blocked. Here, we investigated whether blockade of the terminal 5-HT1B autoreceptor influences a selective 5-HT reuptake inhibitor in the same way, and whether there is an additional effect of blocking both the 5-HT1A and 5-HT1B autoreceptors. 2. Extracellular 5-HT was measured in frontal cortex of the anaesthetized rat by use of brain microdialysis. In vivo extracellular recordings of 5-HT neuronal activity in the dorsal raphe nucleus (DRN) were also carried out. 3. The selective 5-HT reuptake inhibitor, paroxetine (0.8 mg kg-1, i.v.), increased extracellular 5-HT about 2 fold in rats pretreated with the 5-HT1A receptor antagonist, WAY100635. When administered alone neither paroxetine (0.8 mg kg-1, i.v.) nor WAY100635 (0.1 mg kg-1, i.v.) altered extracellular 5-HT levels. 4. Paroxetine (0.8 mg kg-1, i.v.) did not increase 5-HT in rats pretreated with the 5-HT1B/D receptor antagonist, GR127935 (1 mg kg-1, i.v.). GR127935 (1 and 5 mg kg-1, i.v.) had no effect on extracellular 5-HT when administered alone. 5. Interestingly, paroxetine (0.8 mg kg-1, i.v.) caused the greatest increase in 5-HT (up to 5 fold) when GR127935 (1 or 5 mg kg-1, i.v.) was administered in combination with WAY100635 (0.1 mg kg-1, i.v.). Administration of GR127935 (5 mg kg-1, i.v.) plus WAY100635 (0.1 mg kg-1, i.v.) without paroxetine, had no effect on extracellular 5-HT in the frontal cortex. 6. Despite the lack of effect of GR127935 on 5-HT under basal conditions, when 5-HT output was elevated about 3 fold (by adding 1 microM paroxetine to the perfusion medium), the drug caused a dose-related (1 and 5 mg kg-1, i.v.) increase in 5-HT. 7. By itself, GR127935 slightly but significantly decreased 5-HT cell firing in the DRN at higher doses (2.0-5.0 mg kg-1, i.v.), but did not prevent the inhibition of 5-HT cell firing induced by paroxetine. 8. In summary, our results suggest that selective 5-HT reuptake inhibitors may cause a large increase in 5-HT in the frontal cortex when 5-HT autoreceptors on both the somatodendrites (5-HT1A) and nerve terminals (5-HT1B) are blocked. This increase is greater than when either set of autoreceptors are blocked separately. The failure of a 5-HT1B receptor antagonist alone to enhance the effect of the selective 5-HT reuptake inhibitor in our experiments may be related to a lack of tone on the terminal 5-HT1B autoreceptor due to a continued inhibition of 5-HT cell firing. These results are discussed in relation to the use of 5-HT autoreceptor antagonists to augment the antidepressant effect of selective 5-HT reuptake inhibitors.

背景与目标： 1.当抑制体树突状5-HT1A自体受体时，选择性5-羟色胺（5-HT； 5-羟色胺）再摄取抑制剂（SSRIs）导致前脑细胞外5-HT的增加。在这里，我们调查了终端5-HT1B自身受体的阻断作用是否以相同的方式影响选择性5-HT再摄取抑制剂，以及是否存在阻断5-HT1A和5-HT1B自身受体的其他作用。 2.通过使用脑微透析在麻醉的大鼠的额皮质中测量细胞外5-HT。还进行了背缝核（DRN）中5-HT神经元活性的体内细胞外记录。 3.选择性5-HT再摄取抑制剂帕罗西汀（0.8mg kg-1，静脉内）在用5-HT1A受体拮抗剂WAY100635预处理的大鼠中使细胞外5-HT增加约2倍。当单独给药时，帕罗西汀（0.8 mg kg-1，i.v.）和WAY100635（0.1 mg kg-1，i.v.）均未改变细胞外5-HT水平。 4.在用5-HT1B / D受体拮抗剂GR127935（1 mg kg-1，i.v.）预处理的大鼠中，帕罗西汀（0.8 mg kg-1，i.v.）不会增加5-HT。单独给药时，GR127935（1和5 mg kg-1，静脉内）对细胞外5-HT无效。 5.有趣的是，当GR127935（1或5 mg kg-1，iv）与WAY100635（0.1 mg）联合使用时，帕罗西汀（0.8 mg kg-1，iv）引起5-HT的最大增加（最多5倍）。千克-1，iv）。不含帕罗西汀的GR127935（5 mg kg-1，i.v.）加上WAY100635（0.1 mg kg-1，i.v.）的给药对额皮质中的细胞外5-HT没有影响。 6.尽管在基本条件下GR127935对5-HT缺乏影响，但当5-HT产量提高了约3倍时（通过向灌注培养基中添加1 microM帕罗西汀），该药物引起了剂量相关（1和5 mg kg-1，iv）5-HT升高。 7.就其本身而言，GR127935在较高剂量（2.0-5.0 mg kg-1，i.v.）的DRN中略微但显着降低了5-HT细胞的发射，但并未阻止帕罗西汀诱导的5-HT细胞发射的抑制。 8.总而言之，我们的研究结果表明，当在肢体树突（5-HT1A）和神经末梢（5-HT1B）上都存在5-HT自身受体时，选择性5-HT再摄取抑制剂可能会导致额叶皮层5-HT大量增加。被阻止。该增加量大于分别封闭任一组自动受体时的增加量。在我们的实验中，单独使用5-HT1B受体拮抗剂未能增强选择性5-HT再摄取抑制剂的作用可能与由于持续抑制5-HT细胞而导致末端5-HT1B自体受体缺乏音调有关射击。这些结果与使用5-HT自身受体拮抗剂增强选择性5-HT再摄取抑制剂的抗抑郁作用有关。

BACKGROUND & AIMS: Paired helical filaments (PHFs) in the neurofibrillary tangles (NFTs) in Alzheimer's disease (AD) brains are composed of highly phosphorylated isoforms of tau (PHFtau) that fail to bind microtubules (MTs), and the levels of MT-binding competent tau are decreased in AD brains with abundant PHFtau. Because this loss of MT binding could compromise the viability of tangle-bearing AD neurons by destabilizing MTs, we asked whether these events could be initiated by inhibiting protein phosphatase 1 (PP1) and PP2A in cultured human neurons (NT2N cells) using okadaic acid (OK) and calyculin-A (CL-A). The treatment of NT2N cells with OK and CL-A increased tau phosphorylation, decreased the binding of tau to MTs, and selectively depolymerized the more stable detyrosinated MTs but not the more labile tyrosinated MTs. Significantly, this led to the rapid degeneration of axons, which are enriched in the more stable detyrosinated MTs, and PP2A was implicated in the initiation of this cascade of events because PP2A but not PP1 was closely associated with MTs in the NT2N cells. These studies imply that inactivation of PP2A in vulnerable neurons of the AD brain may play a mechanistic role in the conversion of normal tau into PHFtau, in the depolymerization of stable MTs, and in the degeneration of axons emanating from tangle-bearing neurons.

背景与目标： 阿尔茨海默氏病（AD）大脑中神经原纤维缠结（NFT）中的成对螺旋丝（PHF）由无法结合微管（MTs）的高度磷酸化的tau同工型（PHFtau）组成，与MT结合的有效tau的水平为具有丰富的PHFtau的AD大脑减少。由于这种MT结合的丧失可能会通过破坏MT的稳定性来破坏带有缠结的AD神经元的生存能力，因此我们问这些事件是否可以通过使用冈田酸（NT2N细胞）抑制培养的人类神经元（NT2N细胞）中的蛋白磷酸酶1（PP1）和PP2A来引发。 OK）和calyculin-A（CL-A）。用OK和CL-A处理NT2N细胞增加tau磷酸化，降低tau与MT的结合，并选择性解聚更稳定的脱酪氨酸MTs，但不解聚更不稳定的酪氨酸MTs。值得注意的是，这导致轴突快速变性，轴突迅速富集在更稳定的脱酪氨酸MT中，而PP2A参与了这一级联事件的发​​生，因为PP2A而非PP1与NT2N细胞中的MT密切相关。这些研究表明，AD大脑脆弱神经元中PP2A的失活可能在正常tau转化为PHFtau，稳定MT的解聚以及含缠结神经元的轴突变性方面发挥了机械作用。 >

BACKGROUND & AIMS: :Hagfish (Eptatretus burgeri) are agnathous and are the earliest vertebrates still in existence. Pavement cells adjacent to the mitochondria-rich cells show orthogonal arrays of particles (OAPs) in the gill of hagfish, a known ultrastructural morphology of aquaporin (AQP) in mammalian freeze-replica studies, suggesting that an AQP homolog exists in pavement cells. We therefore cloned water channels from hagfish gill and examined their molecular characteristics. The cloned AQP [E. burgeri AQP4 (EbAQP4)] encodes 288 amino acids, including two NPA motifs and six transmembrane regions. The deduced amino acid sequence of EbAQP4 showed high homology to mammalian and avian AQP4 (rat, 44%; quail, 43%) and clustered with AQP4 subsets by the molecular phylogenetic tree. The osmotic water permeability of Xenopus oocytes injected with EbAQP4 cRNA increased eightfold compared with water-injected controls and was not reversibly inhibited by 0.3 mM HgCl(2). EbAQP4 mRNA expression in the gill was demonstrated by the RNase protection assay; antibody raised against the COOH terminus of EbAQP4 also detected (by Western blot analysis) a major approximately 31-kDa band in the gill. Immunohistochemistry and immunoelectron microscopy showed EbAQP4 localized along the basolateral membranes of gill pavement cells. In freeze-replica studies, OAPs were detected on the protoplasmic face of the split membrane comprising particles 5-6 nm long on the basolateral side of the pavement cells. These observations suggest that EbAQP4 is an ancestral water channel of mammalian AQP4 and plays a role in basolateral water transport in the gill pavement cells.

背景与目标： H鱼（Eptatretus burgeri）凶猛，是现存的最早的脊椎动物。与线粒体富集细胞相邻的路面细胞在show鱼的show中显示出正交排列的颗粒（OAP），这是哺乳动物冷冻复制研究中水通道蛋白（AQP）的已知超微结构形态，表明路面细胞中存在AQP同源物。因此，我们从ha鱼g中克隆了水通道，并检查了它们的分子特征。克隆的AQP [E. burger AQP4（EbAQP4）]编码288个氨基酸，包括两个NPA基序和六个跨膜区域。推导的EbAQP4氨基酸序列与哺乳动物和禽类AQP4具有高度同源性（大鼠为44％；鹌鹑为43％），并通过分子系统树与AQP4亚群聚集在一起。与注水对照组相比，注有EbAQP4 cRNA的非洲爪蟾卵母细胞的渗透水渗透率增加了八倍，并且没有被0.3 mM HgCl（2）可逆地抑制。 RNase保护试验证实了ill中EbAQP4 mRNA的表达。 （通过蛋白质印迹分析）还检测到针对EbAQP4的COOH末端的抗体，在ill中有大约31kDa的主要条带。免疫组织化学和免疫电子显微镜检查显示，EbAQP4沿along路面细胞的基底外侧膜定位。在冷冻复制品研究中，在分裂膜的质膜面上检测到OAP，该膜在路面细胞的基底外侧上长5-6 nm的颗粒。这些观察结果表明，EbAQP4是哺乳动物AQP4的祖先水通道，并且在the路面细胞的基底外侧水运输中起作用。

BACKGROUND & AIMS: :Diagnosis of eukaryotic parasitic infection using antibody-based tests such as ELISAs (enzyme-linked immunosorbent assays) is often problematic because of the need to differentiate between homologous host and pathogen proteins and to ensure that antibodies raised against a peptide will also bind to the peptide in the context of its three-dimensional protein structure. Filariasis caused by the nematode, Brugia malayi, is an important worldwide tropical disease in which parasites disappear from the bloodstream during daylight hours, thus hampering standard microscopic diagnostic methods. To address this problem, a structural approach was used to develop monoclonal antibodies (mAbs) that detect asparaginyl-tRNA synthetase (AsnRS) secreted from B. malayi. B. malayi and human AsnRS amino acid sequences were aligned to identify regions that are relatively unconserved, and a 1.9 A crystallographic structure of B. malayi AsnRS was used to identify peptidyl regions that are surface accessible and available for antibody binding. Sequery and SSA (Superpositional Structural Analysis) software was used to analyze which of these peptides was most likely to maintain its native conformation as a synthetic peptide, and its predicted helical structure was confirmed by NMR. A 22-residue peptide was synthesized to produce murine mAbs. Four IgG(1) mAbs were identified that recognized the synthetic peptide and the full-length parasite AsnRS, but not human AsnRS. The specificity and affinity of mAbs was confirmed by Western blot, immunohistochemistry, surface plasmon resonance, and enzyme inhibition assays. These results support the success of structural modeling to choose peptides for raising selective antibodies that bind to the native protein.

背景与目标： ：由于需要区分同源宿主蛋白和病原体蛋白，并确保针对肽的抗体也能与抗体结合，因此使用基于抗体的测试（例如ELISA）（酶联免疫吸附测定）对真核寄生虫感染的诊断通常存在问题。肽在其三维蛋白质结构的背景下。由线虫引起的丝虫病是马来亚布鲁吉（Brugia malayi），是一种重要的世界性热带病，在白天，其寄生虫会从血液中消失，从而妨碍了标准的微观诊断方法。为了解决该问题，使用了一种结构方法来开发单克隆抗体（mAb），该单克隆抗体可检测马来芽孢杆菌分泌的天冬酰胺基-tRNA合成酶（AsnRS）。将马来芽孢杆菌和人AsnRS氨基酸序列进行比对以鉴定相对不保守的区域，并且使用马来芽孢杆菌AsnRS的1.9 A晶体学结构鉴定表面可及且可用于抗体结合的肽基区域。使用Sequery和SSA（叠加结构分析）软件分析这些肽中的哪一种最有可能作为合成肽保持其天然构象，并通过NMR证实了其预测的螺旋结构。合成22个残基的肽以产生鼠mAb。鉴定出四个IgG（1）mAb，它们可识别合成肽和全长寄生虫AsnRS，但不能识别人AsnRS。 mAb的特异性和亲和力已通过Western印迹，免疫组织化学，表面等离振子共振和酶抑制试验得以证实。这些结果支持结构建模成功地选择了肽，以产生与天然蛋白结合的选择性抗体。

BACKGROUND & AIMS: :The purpose of this study was to examine whether selective iNOS inhibition can restore the hemodynamic changes and reduce the nitrotyrosine levels in the cerebral cortex of rats with streptozotocin-induced diabetes during endotoxin-induced shock. The study was designed to include three sets of experiments: (1) measurement of changes in systemic hemodynamics, (2) measurement of biochemical variables, including iNOS activity and nitrotyrosine formation in the brain, and (3) assessment of mortality rate. Rats were randomly divided into four groups: group 1, control; group 2, LPS: Escherichia coli endotoxin, 10.0 mg/kg (i.v.) bolus; group 3 (i.v.) LPS and L-N6-(1-iminoethyl)-lysine (L-NIL), 4mg/kg (i.p.); and group 4, LPS and NG-nitro-L-arginine methyl ester (L-NAME), 5 mg/kg (i.p.). In nondiabetic rats, administration of L-NIL prevented the hemodynamic and biochemical changes, and increases in plasma nitrite and cerebral nitrotyrosine levels induced by LPS. Administration of L-NAME partially prevented these LPS-induced changes. On the other hand, in diabetic rats, administration of L-NIL only partially prevented the hemodynamic and biochemical changes, and increases in plasma nitrite and cerebral nitrotyrosine levels associated with LPS. Administration of L-NAME, however, had no effects on these LPS-induced changes in diabetic rats. There was a significant difference in nitrotyrosine levels between nondiabetic and diabetic rats in groups 2, 3, and 4 at 2 and 3 h after the treatment (at 3 h; nondiabetic--control, 4.6 +/- 0.4; LPS (i.v.), 8.9 +/- 1.0, LPS (i.v.) + L-NIL, 4.7 +/- 0.5; LPS (i.v.) + L-NAME, 7.1 +/- 0.9; diabetic--control, 5.5 +/- 0.4; LPS (i.v.), 13.6 +/- 1.2; LPS (i.v.) + L-NIL, 9.0 +/- 0.9; LPS (i.v.) + L-NAME, 13.0 +/- 1.0; densitometric units). Insulin therapy resulted in a decrease in iNOS activity (at 3 h: 1.0 +/- 0.5 fmol mg min), nitrotyrosine formation (at 3 h; 5.0 +/- 0.5, densitometric units), and mortality rates (30% at 6 h, 50% at 12 h) in the LPS (i.v.) + L-NIL group of diabetic rats. Selective iNOS inhibition in diabetic rats could not improve hemodynamic instability, chemical changes, iNOS activity, and nitrotyrosine formation during septic shock compared with the improvements observed in nondiabetic rats. Tight glucose control along with administration of L-NIL can result in more effective restoration of the biochemical changes of septicemia in diabetic rats. Thus, hyperglycemia may be one of the mechanisms related to the aggravation of endotoxin-induced shock.

背景与目标： ：本研究的目的是研究选择性内源性iNOS抑制能否在内毒素诱导的休克期间恢复链脲佐菌素诱发的糖尿病大鼠的血流动力学变化并降低其大脑皮质的硝基酪氨酸水平。该研究设计为包括三组实验：（1）测量全身血流动力学的变化，（2）测量生化变量，包括iNOS活性和脑中硝基酪氨酸的形成，以及（3）死亡率评估。将大鼠随机分为四组：第1组，对照组；和第2组。第2组，LPS：大肠埃希菌内毒素，每次推注10.0 mg / kg（i.v.）；第3组（静脉）LPS和L-N6-（1-亚氨基乙基）-赖氨酸（L-NIL），4mg / kg（腹膜）;第4组，LPS和NG-硝基-L-精氨酸甲酯（L-NAME），5 mg / kg（腹膜内）。在非糖尿病大鼠中，L-NIL的给药阻止了血流动力学和生化变化，并增加了LPS诱导的血浆亚硝酸盐和脑硝基酪氨酸水平。 L-NAME的管理部分阻止了这些LPS引起的变化。另一方面，在糖尿病大鼠中，L-NIL的施用仅部分阻止了血流动力学和生化变化，并增加了与LPS相关的血浆亚硝酸盐和脑硝基酪氨酸水平。但是，在糖尿病大鼠中，L-NAME的给药对这些LPS诱导的变化没有影响。第2、3和4组的非糖尿病和糖尿病大鼠在治疗后2和3小时的硝酸酪氨酸水平存在显着差异（3小时;非糖尿病对照组为4.6 /-0.4; LPS（iv）为8.9） /-1.0，LPS（iv）L-NIL，4.7 /-0.5； LPS（iv）L-NAME，7.1 /-0.9；糖尿病对照，5.5 /-0.4； LPS（iv），13.6 /-1.2； LPS（iv）L-NIL，9.0 /-0.9； LPS（iv）L-NAME，13.0 /-1.0；光密度单位）。胰岛素治疗导致iNOS活性（3 h：1.0 /-0.5 fmol mg min），硝基酪氨酸形成（3 h; 5.0 /-0.5，光密度单位）和死亡率（6h，50时30％）降低LPS（iv）L-NIL组的糖尿病大鼠在12 h时的％）。与非糖尿病大鼠相比，对糖尿病大鼠的选择性iNOS抑制不能改善败血性休克期间的血流动力学不稳定，化学变化，iNOS活性和硝基酪氨酸形成。严格的葡萄糖控制以及L-NIL的使用可以使糖尿病大鼠败血病的生化变化更有效地恢复。因此，高血糖症可能是与内毒素诱发的休克加重有关的机制之一。

BACKGROUND & AIMS: BACKGROUND:The risk of birth defects after antenatal exposure to selective serotonin-reuptake inhibitors (SSRIs) remains controversial. METHODS:We assessed associations between first-trimester maternal use of SSRIs and the risk of birth defects among 9849 infants with and 5860 infants without birth defects participating in the Slone Epidemiology Center Birth Defects Study. RESULTS:In analyses of defects previously associated with SSRI use (involving 42 comparisons), overall use of SSRIs was not associated with significantly increased risks of craniosynostosis (115 subjects, 2 exposed to SSRIs; odds ratio, 0.8; 95% confidence interval [CI], 0.2 to 3.5), omphalocele (127 subjects, 3 exposed; odds ratio, 1.4; 95% CI, 0.4 to 4.5), or heart defects overall (3724 subjects, 100 exposed; odds ratio, 1.2; 95% CI, 0.9 to 1.6). Analyses of the associations between individual SSRIs and specific defects showed significant associations between the use of sertraline and omphalocele (odds ratio, 5.7; 95% CI, 1.6 to 20.7; 3 exposed subjects) and septal defects (odds ratio, 2.0; 95% CI, 1.2 to 4.0; 13 exposed subjects) and between the use of paroxetine and right ventricular outflow tract obstruction defects (odds ratio, 3.3; 95% CI, 1.3 to 8.8; 6 exposed subjects). The risks were not appreciably or significantly increased for other defects or other SSRIs or non-SSRI antidepressants. Exploratory analyses involving 66 comparisons showed possible associations of paroxetine and sertraline with other specific defects. CONCLUSIONS:Our findings do not show that there are significantly increased risks of craniosynostosis, omphalocele, or heart defects associated with SSRI use overall. They suggest that individual SSRIs may confer increased risks for some specific defects, but it should be recognized that the specific defects implicated are rare and the absolute risks are small.

背景与目标： 背景：产前暴露于选择性5-羟色胺再摄取抑制剂（SSRIs）后出生缺陷的风险仍然存在争议。
方法：我们评估了参加妊娠流行病学中心出生缺陷研究的9849例有和5860例无出生缺陷婴儿的孕早期孕产妇使用SSRI与出生缺陷风险之间的关联。
结果：在先前与SSRI使用相关的缺陷的分析中（涉及42个比较），SSRI的整体使用与颅突狭窄的风险显着增加无关（115名受试者，有2名接触SSRI；优势比为0.8； 95％置信区间[CI] ]，0.2到3.5），全卵裂（127名受试者，暴露3；比值比，1.4； 95％CI，0.4到4.5），或总体上有心脏缺陷（3724名受试者，暴露100，比值比，1.2，95％CI，0.9）至1.6）。对单个SSRI与特定缺陷之间的关联进行的分析显示，使用舍曲林和全卵石（赔率，5.7； 95％CI，1.6至20.7； 3个暴露的受试者）与间隔缺损（赔率，2.0； 95％CI）之间存在显着关联。 ； 1.2至4.0； 13位暴露的受试者）以及在帕罗西汀和右心室流出道梗阻之间的使用（赔率，3.3； 95％CI，1.3至8.8； 6位暴露的受试者）。其他缺陷或其他SSRI或非SSRI抗抑郁药的风险未显着或显着增加。涉及66个比较的探索性分析表明，帕罗西汀和舍曲林可能与其他特定缺陷有关。
结论：我们的研究结果并未表明与SSRI的使用相关的颅突增生，卵泡扩张或心脏缺陷的风险显着增加。他们认为，单个SSRI可能会增加某些特定缺陷的风险，但是应该认识到，所涉及的特定缺陷很少，绝对风险也很小。

BACKGROUND & AIMS: PURPOSE:To elucidate the intraocular pressure (IOP)-lowering effects and associated characteristics of Y-39983, a selective Rho-associated coiled coil-forming protein kinase (ROCK) inhibitor derived from Y-27632, in animal eyes. METHODS:Y-39983 was compared with Y-27632 for selectivity of ROCK inhibition by biochemical assay. The IOP was monitored by pneumatonometer in albino rabbits and cynomolgus monkeys that were given topically administered Y-39983. The total outflow facility and uveoscleral outflow were measured by two-level constant-pressure perfusion and perfusion technique using fluorescein isothiocyanate-dextran, respectively, at 2 hours after topical administration of Y-39983 in albino rabbits. The ocular toxicologic effects of topical administration of Y-39983 were observed in albino rabbits and cynomolgus monkeys. RESULTS:A biochemical assay showed that Y-39983 inhibited ROCK more potently than Y-27632. In rabbits, topical administration of Y-39983 significantly increased conventional outflow by 65.5%, followed by significant, dose-dependent reduction in IOP. Maximum IOP reduction was 13.2 +/- 0.6 mm Hg (mean +/- SE) at 0.1% Y-39983 in rabbits. In monkeys, at 3 hours after topical administration of 0.05% Y-39983, maximum reduction of IOP was 2.5 +/- 0.8 mm Hg. No serious side effects were observed in ocular tissues except sporadic punctate subconjunctival hemorrhage during long-term topical administration of Y-39983 four times a day (at 2-hour intervals) in rabbits or monkeys. However, punctate subconjunctival hemorrhage was not observed with administration twice daily (at a 6-hour interval) or three times a day (at 5-hour intervals). CONCLUSIONS:Y-39983 causes increased outflow facility followed by IOP reduction. Y-39983 ophthalmic solution may be a candidate drug for lowering of IOP, since it increases conventional outflow and produces relatively few side effects.

背景与目标： 目的：为了阐明动物眼中Y-39983（一种选自R-27632的选择性Rho相关的卷曲螺旋形成蛋白激酶（ROCK）抑制剂）Y-39983的降低眼内压（IOP）的作用和相关特征。
方法：通过生化分析比较Y-39983和Y-27632对ROCK的选择性。通过肺气量计在局部给予Y-39983的白化兔和食蟹猴中监测IOP。在白化兔中局部施用Y-39983后2小时，分别通过两级恒压灌注和灌注技术使用异硫氰酸荧光素-右旋糖酐测量总流出设施和葡萄膜巩膜流出。在白化病兔和食蟹猴中观察到局部施用Y-39983的眼部毒理作用。
结果：生化分析表明，Y-39983比ROCK-Y-27632对ROCK的抑制作用更强。在兔子中，局部施用Y-39983可将常规流出量显着增加65.5％，然后是IOP的剂量依赖性显着降低。在0.1％的Y-39983中，兔子的最大IOP降低为13.2 /-0.6毫米汞柱（平均/-SE）。在猴子中，局部给药0.05％Y-39983后3小时，IOP的最大降低为2.5±0.8 mm Hg。在兔子或猴子中，一天四次（以2小时为间隔）每天四次Y-39983长期局部给药期间，偶发性点状结膜下出血除外，在眼组织中未观察到严重的副作用。但是，每日两次（每隔6小时）或每天三次（每隔5小时）给药未观察到点状结膜下出血。
结论：Y-39983引起流出设施增加，随后IOP降低。 Y-39983眼用溶液剂可能是降低IOP的候选药物，因为它增加了常规流出量并且产生了相对较少的副作用。

BACKGROUND & AIMS: :We have used combinatorial chemistry with amino acid mixtures (X) at positions 6 to 23 in vasoactive intestinal peptide (VIP) to optimize binding affinity and selectivity to the rat VPAC(1) receptor. The most efficient amino acid replacement was a substitution of alanine at position 18 to diphenylalanine (Dip), increasing the displacement efficiency of (125)I-VIP by 370-fold. The [Dip(18)]VIP(6-23) was subsequently used to find a second replacement, employing the same approach. Tyrosine at position 9 was selected and the resulting [Tyr(9),Dip(18)]VIP(6-23) analog has a K(i) value of 90 nM. This analog was unable to stimulate cAMP production at 10(-6) M but was able to inhibit VIP-induced cAMP stimulation (K(b) = 79 nM). The K(i) values of [Tyr(9),Dip(18)]VIP(6-23) using the rat VPAC(2) and PAC(1) receptors were 3,000 nM and >10,000 nM, respectively. Thus, [Tyr(9),Dip(18)]VIP(6-23) is a selective VPAC(1) receptor antagonist. The C-terminally extended form, [Tyr(9),Dip(18)]VIP(6-28), displays improved antagonistic properties having a K(i) and K(b) values of 18 nM and 16 nM, respectively. On the contrary, the fully extended form, [Tyr(9),Dip(18)]VIP(1-28), was a potent agonist with improved binding affinity (K(i) = 0.11 nM) and ability to stimulate cAMP (EC(50) = 0.23 nM) compared with VIP (K(i) = 1.7 nM, EC(50) = 1.12 nM). Furthermore, the specificity of this agonist to the VPAC(1) receptor was high, the K(i) values for the VPAC(2) and PAC(1) receptors were 53 nM and 3,100 nM, respectively. Seven other analogs with the [Tyr(9),Dip(18)] replacement combined with previously published VIP modifications have been synthesized and described in this work.

背景与目标： ：我们已将组合化学与血管活性肠肽（VIP）中6至23位的氨基酸混合物（X）结合使用，以优化与大鼠VPAC（1）受体的结合亲和力和选择性。最有效的氨基酸替换是将18位丙氨酸替换为二苯丙氨酸（Dip），使（125）I-VIP的置换效率提高370倍。随后，使用相同的方法使用[Dip（18）] VIP（6-23）查找第二个替换项。选择第9位的酪氨酸，所得的[Tyr（9），Dip（18）] VIP（6-23）类似物的K（i）值为90 nM。该类似物无法在10（-6）M刺激cAMP产生，但能够抑制VIP诱导的cAMP刺激（K（b）= 79 nM）。使用大鼠VPAC（2）和PAC（1）受体的[Tyr（9），Dip（18）] VIP（6-23）的K（i）值分别为3,000 nM和> 10,000 nM。因此，[Tyr（9），Dip（18）] VIP（6-23）是选择性的VPAC（1）受体拮抗剂。 C末端扩展形式[Tyr（9），Dip（18）] VIP（6-28）显示出改善的拮抗特性，其K（i）和K（b）值分别为18 nM和16 nM。相反，完全延伸的形式[Tyr（9），Dip（18）] VIP（1-28）是一种有效的激动剂，具有改善的结合亲和力（K（i）= 0.11 nM）和刺激cAMP的能力（ EC（50）= 0.23 nM），而VIP（K（i）= 1.7 nM，EC（50）= 1.12 nM）。此外，该激动剂对VPAC（1）受体的特异性很高，VPAC（2）和PAC（1）受体的K（i）值分别为53 nM和3,100 nM。在这项工作中，已经合成并描述了七个其他具有[Tyr（9），Dip（18）]替代物并结合了先前发布的VIP修饰的类似物。

BACKGROUND & AIMS: :The purpose of this study was to assess 4 Fr Judkins catheters with modified shorter tips for performing selective coronary angiography in infants and young children. Twenty patients ranging 6 weeks to 3. 8 years of age were enrolled. Retrograde left heart catheterization and selective coronary angiography were performed. Right (JR) and left (JL) catheters with modified 1.5- and 2.5-cm curves (Cordis) were used. Thirty-six of 37 coronary arteries were successfully cannulated and demonstrated. Median procedure time was 95 sec for the right and 50 sec for the left coronary artery. Median fluoroscopy time was 1.1 min for the right and 0.7 min for the left coronary artery. The JL 1.5 appeared best suited for patients less than 75 cm tall. The JR 1.5 was suitable for patients up to 85 cm tall. Taller patients required the 2.5-cm curves. It is concluded that these modified 4 Fr Judkins catheters were effective.

背景与目标： ：本研究的目的是评估4个Fr Judkins导管，这些导管具有改良的较短尖端，用于在婴幼儿中进行选择性冠状动脉造影。招募了20名6周至3. 8岁的患者。进行逆行左心导管检查和选择性冠状动脉造影。使用具有改良的1.5厘米和2.5厘米曲线的右（JR）和左（JL）导管（Cordis）。 37例冠状动脉中有36例成功插管并显示。右侧冠状动脉的中位手术时间为95秒，左侧冠状动脉的中位手术时间为50秒。透视的中位时间，右侧冠状动脉为1.1分钟，左侧冠状动脉为0.7分钟。 JL 1.5似乎最适合身高不到75厘米的患者。 JR 1.5适用于身高不超过85厘米的患者。较高的患者需要2.5厘米的曲线。结论是这些改良的4 Fr Judkins导管是有效的。

BACKGROUND & AIMS: 1. The actions of a chromatographically identified extract of the marine dinoflagellate Ostreopsis lenticularis, named ostreotoxin-3 (OTX-3), were studied on frog isolated neuromuscular preparations. 2. OTX-3 (1-10 microg ml(-1)) applied to cutaneous pectoris nerve-muscle preparations depolarized skeletal muscle fibres and caused spontaneous contractions. The depolarization was neither reversed by prolonged washing nor by (+)-tubocurarine. 3. OTX-3 decreased the amplitude of miniature end plate potentials (m.e.p.ps) but did not affect their frequency. 4. Extracellular recording of compound action potentials revealed that OTX-3 affected neither excitability nor conduction along intramuscular nerve branches. 5. End-plate potentials (e.p.ps) elicited by nerve stimulation were reduced in amplitude by OTX-3 and even showed reversed polarity in junctions deeply depolarized by the toxin. 6. Membrane depolarization induced by OTX-3 was decreased about 70% in muscles pretreated for 30 min with 10 microM tetrodotoxin. In contrast, muscles pretreated with 5 microM mu-conotoxin GIIIA were completely insensitive to OTX-3-induced depolarization. 7. OTX-3 did not affect e.p.p. amplitude and the quantal content of e.p.ps in junctions in which muscle depolarization was abolished by mu-conotoxin GIIIA. 8. OTX-3 is a novel type of sodium-channel activating toxin that discriminates between nerve and skeletal muscle membranes.

背景与目标： 1.在蛙类分离的神经肌肉制剂上研究了一种色谱鉴定的海洋鞭毛鞭毛骨癣菌提取物，称为ostreotoxin-3（OTX-3）的作用。 2. OTX-3（1-10微克ml（-1））应用于皮肤胸大肌神经肌肉制剂，可使骨骼肌纤维去极化并引起自发性收缩。长时间洗涤或（）-微管尿素均不能逆转去极化作用。 3. OTX-3减小了微型端板电位的幅度（m.e.p.ps），但不影响其频率。 4.复合动作电位的细胞外记录显示，OTX-3既不影响兴奋性也不影响沿肌内神经分支的传导。 5.神经刺激引起的终板电位（e.p.ps）被OTX-3降低，甚至在被毒素深层去极化的连接处显示相反的极性。 6.在用10 microM河豚毒素预处理30分钟的肌肉中，由OTX-3诱导的膜去极化减少了约70％。相反，用5 microM mu-conotoxin GIIIA预处理的肌肉对OTX-3诱导的去极化完全不敏感。 7. OTX-3不会影响e.p.p. mu-conotoxin GIIIA消除了肌肉去极化的连接处的振幅和e.p.ps的含量。 8. OTX-3是一种新型的钠通道激活毒素，可区分神经膜和骨骼肌膜。