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: INTRODUCTION:Adverse drug reactions (ADRs) occur frequently in hospitals and increase costs of health care; however, few studies have quantified the clinical and economic impact of ADRs in Colombia. OBJECTIVES:These impacts were evaluated by calculating costs associated with ADRs in patients hospitalized in the internal medicine ward of a Level 3 hospital located in Bogotá, Colombia. In addition, salient clinical features of ADRs were identified and characterized. MATERIAL AND METHODS:Intensive follow-ups for a cohort of patients were conducted for a five month period in order to detect ADRs; different ways to classify them, according to literature, were considered as well. Information was collected using the INVIMA reporting format, and causal probability was evaluated with the Naranjo algorithm. Direct costs were calculated from the perspective of payer, based on the following costs: additional hospital stay, medications, paraclinical tests, additional procedures, patient displacement to intermediate or intensive care units, and other costs. RESULTS:Of 836 patients admitted to the service, 268 adverse drug reactions were detected in 208 patients (incidence proportion 25.1%, occurence rate 0.32). About the ADRs found, 74.3% were classified as probable, 92.5% were type A, and 81.3% were moderate. The body system most often affected was the circulatory system (33.9%). Drugs acting on the blood were most frequently those ones associated with adverse reactions (37.6%). The costs resulting from medical care of adverse drug reactions varied from COL dollar 93,633,422 (USD dollar 35,014.92) to COL dollar 122,155,406 (USD dollar 45,680.94), according to insurance type, during the study period. CONCLUSIONS:Adverse drug reactions have a significant negative health and financial impact on patient welfare. Because of the substantial resources required for their medical care and the significant proportion of preventable adverse reactions, active programs of institutional pharmacovigilance are highly recommended.

背景与目标： 简介：药物不良反应（ADR）在医院中经常发生，并增加了医疗保健费用；但是，很少有研究能够量化ADR在哥伦比亚的临床和经济影响。
目的：通过计算在哥伦比亚波哥大三级医院内科病房住院的患者与ADR相关的费用来评估这些影响。此外，ADR的显着临床特征已得到鉴定和表征。
材料与方法：对一组患者进行了为期五个月的强化随访，以检测ADR。根据文献，还考虑了不同的分类方法。使用INVIMA报告格式收集信息，并使用Naranjo算法评估因果概率。直接费用是根据以下费用从付款人的角度计算的：额外的住院时间，药物，辅助临床检查，其他程序，将患者转移到中级或重症监护室以及其他费用。
结果：836例入院患者中，208例患者发生了268例药物不良反应（发生率25.1％，发生率0.32）。关于发现的ADR，有74.3％被归为可能，A型为92.5％，中度为81.3％。受影响最严重的身体系统是循环系统（33.9％）。作用于血液的药物最常见于那些与不良反应有关的药物（37.6％）。根据保险类型，在研究期间，药物不良反应的医疗费用从93,633,422美元（35,014.92美元）到122,155,406美元（45,680.94美元）不等。
结论：药物不良反应会对患者的健康产生重大的负面健康和财务影响。由于其医疗服务所需的大量资源以及可预防的不良反应的比例很大，因此强烈建议积极进行机构药物警戒计划。

BACKGROUND & AIMS: The GATA family of vertebrate DNA binding regulatory proteins are expressed in diverse tissues and at different times of development. However, the DNA binding regions of these proteins possess considerable homology and recognize a rather similar range of DNA sequence motifs. DNA binding is mediated through two domains, each containing a zinc finger. Previous results have led to the conclusion that although in some cases the N-terminal finger can contribute to specificity and strength of binding, it does not bind independently, whereas the C-terminal finger is both necessary and sufficient for binding. Here we show that although this is true for the N-terminal finger of GATA-1, those of GATA-2 and GATA-3 are capable of strong independent binding with a preference for the motif GATC. Binding requires the presence of two basic regions located on either side of the N-terminal finger. The absence of one of these near the GATA-1 N-terminal finger probably accounts for its inability to bind. The combination of a single finger and two basic regions is a new variant of a motif that has been previously found in the binding domains of other finger proteins. Our results suggest that the DNA binding properties of the N-terminal finger may help distinguish GATA-2 and GATA-3 from GATA-1 and the other GATA family members in their selective regulatory roles in vivo.

背景与目标： GATA家族的脊椎动物DNA结合调节蛋白在不同的组织中以及在不同的发育时期表达。但是，这些蛋白质的DNA结合区具有相当的同源性，可以识别相当相似范围的DNA序列基序。 DNA结合是通过两个结构域介导的，每个结构域都包含一个锌指。先前的结果得出的结论是，尽管在某些情况下N末端指可以有助于结合的特异性和强度，但它不能独立地结合，而C末端指对于结合既是必需的又是足够的。在这里，我们表明，尽管对于GATA-1的N端手指来说确实如此，但GATA-2和GATA-3的手指却能够强烈独立地结合，并优先选择基序GATC。结合需要在N末端指形物的两侧都存在两个基本区域。在GATA-1 N端手指附近缺少这些手指之一可能是其无法结合的原因。单个手指和两个基本区域的组合是一个基序的新变体，该变体先前已在其他手指蛋白的结合域中发现。我们的结果表明，N末端手指的DNA结合特性可能有助于区分GATA-2和GATA-3与GATA-1和其他GATA家族成员在体内的选择性调节作用。

BACKGROUND & AIMS: :Neurons within each layer of cerebral cortex express multiple members of the neurotrophin family and their corresponding receptors. This multiplicity could provide functional redundancy; alternatively, different neurotrophins may direct distinct aspects of cortical neuronal growth and differentiation. By neutralizing endogenous neurotrophins in organotypic slices of developing cortex with Trk receptor bodies (Trk-IgGs), we found that BDNF and NT-3 oppose one another in regulating the dendritic growth of pyramidal neurons. In layer 4, both endogenous and exogenous NT-3 inhibited the dendritic growth stimulated by BDNF. In contrast, in layer 6 both endogenous and exogenous BDNF inhibited dendritic growth stimulated by NT-3. These antagonistic actions of endogenous BDNF and NT-3 provide a mechanism by which dendritic growth and retraction can be dynamically regulated during cortical development, and suggest that the multiple neurotrophins expressed in developing cortex represent distinct components of an extracellular signaling system for regulating dendritic growth.

背景与目标： 大脑皮层每一层中的神经元表达神经营养蛋白家族的多个成员及其相应的受体。这种多样性可以提供功能冗余。或者，不同的神经营养蛋白可指导皮质神经元生长和分化的不同方面。通过用Trk受体体（Trk-IgG）中和发育中的皮质器官的典型切片中的内源性神经营养蛋白，我们发现BDNF和NT-3在调节锥体神经元的树突状生长中彼此相对。在第4层中，内源性NT-3和外源性NT-3均抑制BDNF刺激的树突状生长。相反，在第6层中，内源性和外源性BDNF均抑制NT-3刺激的树突状生长。内源性BDNF和NT-3的这些拮抗作用提供了一种机制，通过该机制可以在皮质发育过程中动态调节树突状细胞的生长和收缩，并表明在发育中的皮质中表达的多种神经营养蛋白代表了调节树突状细胞生长的细胞外信号系统的不同组成部分。

BACKGROUND & AIMS: :Matrix metalloproteinases (MMPs) have been the subject of intense research because of their roles in tumor metastasis and in the rise and spread of degenerative diseases such as osteo- and rheumatoid arthritis. A preliminary class of 140 druglike, small-molecule matrix metalloproteinase-3 inhibitors, intended as starting scaffolds for optimization and synthesis, has been designed in silico using a series of highly predictive three-dimensional quantitative structure-activity relationship models, including comparative molecular field analysis and comparative molecular similarity indices analysis, with docking and scoring. Thalidomide was chosen as the skeleton on which to base the new lead series, as it moderately inhibits MMP-3, is antiangiogenic, and lends itself easily to structural modifications. Most of the new compounds demonstrate medium to high predicted biological activity and good bioavailability as estimated by the octanol-water partition coefficient ClogP. Compound 102 in particular exhibits extremely favorable predicted activity against MMP-3; is moderately bioavailable; satisfies Lipinski's Rule of Five; and shows promise for further optimization, synthesis, and experimental evaluation as a potential adjunct anticancer or antirheumatic therapeutic.

背景与目标： ：基质金属蛋白酶（MMP）由于其在肿瘤转移中以及在变性疾病（例如骨和类风湿性关节炎）的发生和扩散中的作用而受到了广泛的研究。已在计算机上使用一系列高度预测的三维定量结构-活性关系模型（包括比较分子场）在计算机上设计了初步的140类药物样小分子基质金属蛋白酶3抑制剂，旨在作为优化和合成的起始支架。分析和比较分子相似性指数分析，以及对接和评分。选择沙利度胺作为新铅系列的基础，因为它能适度抑制MMP-3，具有抗血管生成作用，并易于进行结构修饰。正如辛醇-水分配系数ClogP所估计的那样，大多数新化合物都具有中等至较高的预测生物活性和良好的生物利用度。特别地，化合物102对MMP-3表现出非常有利的预测活性。具有中等生物利用度；满足Lipinski的五法则；并有望进一步优化，合成和进行实验评估，作为潜在的辅助抗癌药或抗风湿药。

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: :Our recent study demonstrated that defects in p110delta result in B-cell immunodeficiency that is very similar to that observed in BTK-deficient mice. We revealed that the p110delta fit the B-cell signal transduction complex and played a non-redundant role in the development and function of B cells. In humans, most children with primary B-cell immunodeficiency have mutations in the BTK, whereas a few have defects in the components of the B-cell signal transduction complex. But little is known about the genetic variation of p110delta in children with defects in B-cell immunodeficiency of unknown aetiology. Sixteen patients from 15 unrelated families and 112 normal controls underwent sequence analysis to identify genetic variations of the p110delta. Allele frequency in each group was also analysed and compared. We identified five single base-pair polymorphic nucleotide exchanges in both patient and control groups with similar allele frequencies, which did not contribute to the immunodeficiency. Three of them are novel (m.953A>G, m.1200C>T and m.1561A>G), and the m.953A>G and m.1561A>G nucleotide exchanges are non-synonymous (N253S and T456A, respectively). The novel m.1561A>G was in complete linkage disequilibrium with the known m.873A>G in our study of Taiwanese group. In addition, one novel single base-pair missense mutation, m.3256G>A (E1021K), was identified in one boy with typical clinical features of primary B-cell immunodeficiency and could not be found in either his family or the normal control population. By atomic structural analysis of the amino acid as well as the alignment comparison between species, it resulted in the replacement of the negative-charged amino acid E with the positive-charged amino acid K at codon 1021, located in the highly conservative and important catalytic functional domain. Our findings could shed light on further understanding the polymorphisms of p110delta in B-cell immunodeficiency and different populations. Moreover, the 3256G>A missense mutation raised the attention and warranted further extensive analysis to elucidate the role of p110delta in human immunodeficiency.

背景与目标： ：我们最近的研究表明，p110delta中的缺陷会导致B细胞免疫缺陷，这与在BTK缺陷小鼠中观察到的缺陷非常相似。我们发现p110delta适合B细胞信号转导复合物，并且在B细胞的发育和功能中起着非冗余的作用。在人类中，大多数患有原发性B细胞免疫缺陷的儿童在BTK中都有突变，而少数儿童在B细胞信号转导复合物中的成分存在缺陷。但是对于病因不明的B细胞免疫缺陷的儿童p110δ的遗传变异知之甚少。来自15个无关家庭和112个正常对照的16位患者接受了序列分析，以鉴定p110delta的遗传变异。还对每组中的等位基因频率进行了分析和比较。我们在患者和对照组中确定了五个具有相似等位基因频率的单碱基​​对多态性核苷酸交换，这对免疫缺陷没有帮助。其中三个是新颖的（m.953A> G，m.1200C> T和m.1561A> G），而m.953A> G和m.1561A> G核苷酸交换是非同义的（分别为N253S和T456A） ）。在我们对台湾人的研究中，小说m.1561A> G与已知的m.873A> G处于完全连锁不平衡状态。此外，在一个男孩中发现了一个新颖的单碱基对错义突变，m.3256G> A（E1021K），该男孩具有原发性B细胞免疫缺陷的典型临床特征，在其家人或正常对照人群中均未发现。通过氨基酸的原子结构分析以及物种之间的比对比较，结果发现位于高度保守且重要的催化分子1021密码子上的负电荷氨基酸E被正电荷氨基酸K取代。功能域。我们的发现可能有助于进一步了解B细胞免疫缺陷和不同人群中p110delta的多态性。此外，3256G> A错义突变引起了人们的注意，并需要进行进一步的广泛分析以阐明p110delta在人类免疫缺陷中的作用。

BACKGROUND & AIMS: :Epithelial cells play an important role in orchestrating mucosal immune responses. In allergic-type inflammation, epithelial cells control the recruitment of eosinophils into the mucosa. Th2-type cytokine-driven release of eosinophil-active chemokines from epithelial cells directs eosinophil migration into the mucosal epithelium. CCR3, the main eosinophil chemokine receptor, regulates this process; however, the respective contribution of individual CCR3 ligands in eosinophil transepithelial migration is less well understood. Using an in vitro transepithelial chemotaxis system, we found that eotaxin-3 produced by IL-4-stimulated airway epithelial cells and CCR3 on eosinophils exclusively mediate eosinophil transepithelial migration. Eotaxin-3 protein levels were also increased in the nasal mucosal epithelium recovered from allergic patients as compared to non-allergic patients. Surprisingly, eotaxin-3 in IL-4-stimulated airway epithelial cells was predominantly cell surface bound, and the cell surface form was critical for eosinophil transepithelial migration. Eotaxin-3 cell surface association was partially glycosaminoglycan (GAG) dependent, but was completely protein dependent, suggesting that eotaxin-3 associates with both GAG and cell surface proteins. We thus provide evidence that cell surface-associated eotaxin-3 is the critical IL-4-dependent chemotactic signal mediating eosinophil transepithelial migration in the setting of allergic inflammation.

背景与目标： ：上皮细胞在协调粘膜免疫反应中起重要作用。在过敏型炎症中，上皮细胞控制嗜酸性粒细胞向粘膜的募集。 Th2型细胞因子驱动的嗜酸性粒细胞活性趋化因子从上皮细胞的释放引导嗜酸性粒细胞迁移到粘膜上皮中。 CCR3是主要的嗜酸性粒细胞趋化因子受体，调节这一过程。然而，单个CCR3配体在嗜酸性粒细胞上皮细胞迁移中的各自作用尚不清楚。使用体外经上皮趋化系统，我们发现由IL-4刺激的气道上皮细胞和嗜酸性粒细胞上的CCR3产生的eotaxin-3专门介导嗜酸性粒细胞跨上皮迁移。与非过敏患者相比，从过敏患者中回收的鼻粘膜上皮中的Eotaxin-3蛋白水平也有所增加。出人意料的是，IL-4刺激的气道上皮细胞中的eotaxin-3主要与细胞表面结合，并且细胞表面形式对于嗜酸性粒细胞跨上皮迁移至关重要。 Eotaxin-3细胞表面缔合部分依赖糖胺聚糖（GAG），但完全依赖蛋白质，这表明eotaxin-3与GAG和细胞表面蛋白缔合。因此，我们提供证据，在过敏性炎症的情况下，细胞表面相关的eotaxin-3是介导嗜酸性粒细胞跨上皮迁移的关键IL-4依赖性趋化信号。

BACKGROUND & AIMS: Low intensity ultrasound signals, similar to that employed in clinical therapy, are found to mediate differential gene transfer and expression of the Green Fluorescence Protein (GFP) reporter in two human prostate cancer cell lines, LnCap and PC-3. Cell suspensions in the presence or in the absence of GFP (44.5nM) were treated at 37 degrees C under a standing wave condition. Cells were exposed to either continuous wave, 932.7kHz ultrasound, or to several independent bursts, each burst comprising a 20% duty cycle (932.7kHz) sine wave. The burst "repetition" frequency was varied from 10Hz to 10kHz in several different experiments and each treatment received a net identical ultrasound energy exposure. Transient GFP expression levels in viable cells were monitored by flow cytometry. The findings revealed a strong ultrasound tone-burst frequency dependence on the transfection efficiencies. Interestingly, the ultrasound signal parameters which are routinely employed in clinical therapy did not yield any statistically significant enhancement in transfection efficiency relative to their sham counterparts.

背景与目标： 发现低强度超声信号类似于临床治疗中使用的信号，可在两种人前列腺癌细胞系LnCap和PC-3中介导差异基因转移和绿色荧光蛋白（GFP）报告基因的表达。将存在或不存在GFP（44.5nM）的细胞悬浮液在37°C的驻波条件下进行处理。细胞暴露于连续波，932.7kHz超声或几个独立的脉冲串中，每个脉冲串包含20％占空比（932.7kHz）的正弦波。在几个不同的实验中，猝发的“重复”频率在10Hz到10kHz之间变化，并且每种处理都受到净相同的超声能量暴露。通过流式细胞术监测存活细胞中的瞬时GFP表达水平。这些发现揭示了超声音-猝发频率对转染效率的强烈依赖性。有趣的是，相对于假手术，常规用于临床治疗的超声信号参数在转染效率上没有产生任何统计学上的显着提高。

BACKGROUND & AIMS: OBJECTIVE:The scholarly dissemination of innovative medical education practices helps broaden the reach of this type of work, allowing scholarship to have an impact beyond a single institution. There is little guidance in the literature for those seeking to publish program evaluation studies and innovation papers. This study aims to derive a set of evidence-based features of high-quality reports on innovations in emergency medicine (EM) education. METHODS:We conducted a scoping review and thematic analysis to determine quality markers for medical education innovation reports, with a focus on EM. A search of MEDLINE, EMBASE, ERIC, and Google Scholar was augmented by a hand search of relevant publication guidelines, guidelines for authors, and website submission portals from medical education and EM journals. Study investigators reviewed the selected articles, and a thematic analysis was conducted. RESULTS:Our search strategy identified 14 relevant articles from which 34 quality markers were extracted. These markers were grouped into seven important themes: goals and need for innovation, preparation, innovation development, innovation implementation, evaluation of innovation, evidence of reflective practice, and reporting and dissemination. In addition, multiple outlets for the publication of EM education innovations were identified and compiled. CONCLUSION:The publication and dissemination of innovations are critical for the EM education community and the training of health professionals. We anticipate that our list of innovation report quality markers will be used by EM education innovators to support the dissemination of novel educational practices.

背景与目标： 目的：对医学创新实践的学术传播有助于拓宽此类工作的范围，使奖学金的影响力超越单一机构。对于那些寻求发布程序评估研究和创新论文的人，文献中几乎没有指导。这项研究旨在得出一组有关急诊医学教育创新的高质量报告的基于证据的特征。
方法：我们进行了范围审查和主题分析，以确定医学教育创新报告的质量标志，重点是EM。通过手动搜索相关的出版指南，作者指南以及医学教育和EM期刊的网站提交门户，扩大了对MEDLINE，EMBASE，ERIC和Google Scholar的搜索。研究调查人员审查了选定的文章，并进行了主题分析。
结果：我们的搜索策略确定了14篇相关文章，从中提取了34篇质量标记。这些标记分为七个重要主题：创新的目标和需求，准备，创新发展，创新实施，创新评估，反思性实践的证据以及报告和传播。此外，还确定并汇编了多个出版新兴市场教育创新的渠道。
结论：创新的出版和传播对新兴市场教育界和卫生专业人员的培训至关重要。我们期望EM教育创新者将使用我们的创新报告质量标记列表来支持新颖教育实践的传播。

BACKGROUND & AIMS: BACKGROUND AND PURPOSE:Acyl derivatives of CoA have been shown to act as antagonists at human platelet and recombinant P2Y1 receptors, but little is known about their effects in the cardiovascular system. This study evaluated the effect of these endogenous nucleotide derivatives at P2Y1 receptors natively expressed in rat and porcine blood vessels. EXPERIMENTAL APPROACH:Isometric tension recordings were used to evaluate the effects of CoA, acetyl CoA, palmitoyl CoA (PaCoA) and 3'-dephospho-palmitoyl-CoA on concentration relaxation-response curves to ADP and uridine triphosphate (UTP). A FlexStation monitored ADP- and UTP-evoked calcium responses in HEK293 cells. KEY RESULTS:Acetyl CoA and PaCoA, but not CoA, inhibited endothelium-dependent relaxations to ADP with apparent selectivity for P2Y1 receptors (over P2Y(2/4) receptors) in rat thoracic aorta; PaCoA was more potent than acetyl CoA (331-fold vs. fivefold shift of ADP response curve evoked by 10 μM PaCoA and acetyl CoA, respectively); the apparent pA2 value for PaCoA was 6.44. 3'-dephospho-palmitoyl-CoA (10 μM) was significantly less potent than PaCoA (20-fold shift). In porcine mesenteric arteries, PaCoA and the P2Y1 receptor antagonist MRS2500 blocked ADP-mediated endothelium-dependent relaxations; in contrast, they were ineffective against ADP-mediated endothelium-independent relaxation in porcine coronary arteries (which does not involve P2Y1 receptors). Calcium responses evoked by ADP activation of endogenous P2Y1 receptors in HEK293 cells were inhibited in the presence of PaCoA, which failed to alter responses to UTP (acting at endogenous P2Y(2/4) receptors). CONCLUSIONS AND IMPLICATIONS:Acyl derivatives of CoA can act as endogenous selective antagonists of P2Y1 receptors in blood vessels, and this inhibitory effect critically depends on the palmitate and 3'-ribose phosphate substituents on CoA.

背景与目标： 背景和目的：CoA的酰基衍生物已显示出可作为人类血小板和重组P2Y1受体的拮抗剂，但对其在心血管系统中的作用知之甚少。这项研究评估了这些内源核苷酸衍生物对大鼠和猪血管中天然表达的P2Y1受体的作用。
实验方法：使用等距张力记录来评估CoA，乙酰基CoA，棕榈酰CoA（PaCoA）和3'-去磷酸-棕榈酰-CoA对ADP和尿苷三磷酸（UTP）的浓度松弛响应曲线的影响。 FlexStation监视HEK293细胞中ADP和UTP引起的钙反应。
关键结果：乙酰辅酶A和PaCoA，而不是辅酶A，抑制大鼠胸主动脉中P2Y1受体（超过P2Y（2/4）受体）对ADP的内皮依赖性舒张作用，且具有明显的选择性。 PaCoA比乙酰辅酶A更有效（分别由10μMPaCoA和乙酰辅酶A引起的ADP响应曲线的331倍对五倍变化）； PaCoA的表观pA2值为6.44。 3'-去磷酸-棕榈酰基-CoA（10μM）的效力显着低于PaCoA（20倍偏移）。在猪肠系膜动脉中，PaCoA和P2Y1受体拮抗剂MRS2500阻断了ADP介导的内皮依赖性舒张。相反，它们对猪冠状动脉（不涉及P2Y1受体）中ADP介导的内皮依赖性舒张无效。在PaCoA的存在下，HEK293细胞内源性P2Y1受体的ADP激活引起的钙反应被抑制，而PaCoA却不能改变对UTP的反应（作用于内源性P2Y（2/4）受体）。
结论和意义：CoA的酰基衍生物可以作为血管内P2Y1受体的内源性选择性拮抗剂，这种抑制作用主要取决于CoA上的棕榈酸酯和3'-核糖磷酸取代基。

BACKGROUND & AIMS: :Novel double-capped triplet drugs, which have one pharmacophore unit and two epoxymethano or dimethylepoxymethano structures (termed cap or diMe-cap structures, respectively) were synthesized. Key intermediate oxazoline 16 derived from acetone enabled the effective synthesis of double-capped triplets. SYK-134 (7a) and SYK-135 (8a) with N-cyclopropylmethyl substituent and cap structures showed selectivities for the κ opioid receptor. On the other hand, the N-Me series exhibited selectivities for the μ opioid receptor. The double-capped triplet drugs with diMe-cap structures preferred the μ receptor independently of their N-substituents. SYK-385 (19b), one of the μ-selective double-capped triplet drugs, showed the highest selectivity for the μ receptor among the reported μ-selective nonpeptide ligands.

背景与目标： 合成了具有一个药效团单元和两个环氧亚甲基或二甲基环氧亚甲基结构（分别称为帽或二甲基-帽结构）的新型双封端三联体药物。源自丙酮的关键中间体恶唑啉16能够有效合成双封三胞胎。具有N-环丙基甲基取代基和帽结构的SYK-134（7a）和SYK-135（8a）对κ阿片受体具有选择性。另一方面，N-Me系列对μ阿片受体具有选择性。具有diMe-cap结构的双封端三联体药物优选μ受体，而与它们的N取代基无关。微米选择性双封端三联体药物之一SYK-385（19b）在已报道的微米选择性非肽配体中显示出对微米受体的最高选择性。

BACKGROUND & AIMS: :Accumulating evidence indicates that Toll-like receptor (TLR) signaling adapter protein interactions with Toll/Interleukin-1 Receptor (TIR) domains present in sensory neurons may modulate neuropathic pain states. Following ligand interaction with TLRs, TIR serves to both initiate intracellular signaling and facilitate recruitment of signaling adapter proteins to the intracytoplasmic domain. Although TLR TIR is central to a number of TLR signaling cascades, its role in sensory neurons is poorly understood. In this study we investigated the degree to which TLR TIR decoy peptide modified to include a TAT sequence (Trans-Activator of Transcription gene in HIV; TAT-4BB) affected LPS-induced intracellular calcium flux and excitation in sensory neurons, and behavioral changes due to TLR4 active metabolite, morphine-3-glucuronide (M3G) exposure in vivo. TAT-4BB inhibited LPS-induced calcium changes in a majority of sensory neurons and decreased LPS-dependent neuronal excitability in small diameter neurons. Acute systemic administration of the TAT-4BB reversed M3G-induced tactile allodynia in a dose-dependent manner but did not affect motor activity, anxiety or responses to noxious thermal stimulus. These data suggest that targeting TLR TIR domains may provide novel pharmacological targets to reduce or reverse TLR4-dependent pain behavior in the rodent.

背景与目标： ：越来越多的证据表明，Toll样受体（TLR）信号转接头蛋白与感觉神经元中存在的Toll / Interleukin-1受体（TIR）域相互作用可能会调节神经性疼痛状态。配体与TLR相互作用后，TIR既可以启动细胞内信号传导，也可以促进信号传导衔接子蛋白募集到胞质内域。尽管TLR TIR是许多TLR信号级联的核心，但对其在感觉神经元中的作用知之甚少。在这项研究中，我们调查了TLR TIR诱饵肽修饰为包含TAT序列（HIV转录基因的反式激活因子； TAT-4BB）在多大程度上影响LPS诱导的细胞内钙通量和感觉神经元的兴奋，以及行为改变对TLR4活性代谢产物吗啡-3-葡糖醛酸（M3G）的体内暴露。 TAT-4BB抑制了LPS诱导的大多数感觉神经元中钙的变化，并降低了小直径神经元中LPS依赖性神经元的兴奋性。 TAT-4BB的急性全身给药以剂量依赖的方式逆转了M3G诱导的触觉异常性疼痛，但并未影响运动活动，焦虑或对有害热刺激的反应。这些数据表明，靶向TLR TIR结构域可提供新的药理学靶标，以减少或逆转啮齿动物中TLR4依赖性的疼痛行为。

BACKGROUND & AIMS: :Doxazosin used in benign prostatic hyperplasia has the side effects of causing hypotension and the risk of heart failure. The 3 targets of α(1A)-adrenoceptors (in the prostate), α(1D)-adrenoceptors (in the aorta), and an unknown mechanism (in the heart) are involved, respectively. We hypothesized that there is a chiral recognition of doxazosin enantiomers in the 3 targets. Using isolated rat aorta (α(1D)-adrenoceptors) and rabbit prostate (α(1A)-adrenoceptors), we examined pA(2) and pK(B) values of doxazosin enantiomers. We observed chronotropic and inotropic effects of doxazosin enantiomers in isolated rat and rabbit heart tissues. (-)Doxazosin and (+)doxazosin produced a shift to the right of concentration-contraction curves for noradrenalin (aorta) and phenylephrine (prostate smooth muscle). The pA(2) value of (-)doxazosin (8.625 ± 0.053) was smaller than (+)doxazosin (9.503 ± 0.051) in rat aorta, but their pK(B) values in rabbit prostate were the same. In rat and rabbit heart tissues, (+)doxazosin (3-30 µmol·L(-1)) significantly decreased atrial rate, and produced negative inotropic effects; however, (-)doxazosin did not affect the atrial rate, and produced positive inotropic effects in the atria. Thus, the chiral carbon atom of doxazosin does not affect its activity at the therapeutic target of α(1A)-adrenoceptors in the prostate, but significantly changes its blocking activity against α(1D)-adrenoceptors in the aorta, and produces opposite inotropic effects in the atria via an α(1)-adrenoceptor-independent mechanism.

背景与目标： ：多沙唑嗪用于前列腺增生症具有引起低血压和心力衰竭风险的副作用。 α（1A）-肾上腺素受体（在前列腺中），α（1D）-肾上腺素受体（在主动脉中）和未知机制（在心脏中）的3个靶点分别涉及。我们假设在3个靶标中有手性识别多沙唑嗪对映体。使用分离的大鼠主动脉（α（1D）-肾上腺素受体）和兔前列腺（α（1A）-肾上腺素受体），我们检查了多沙唑嗪对映体的pA（2）和pK（B）值​​。我们观察到了多沙唑嗪对映异构体在离体大鼠和兔心脏组织中的变时性和变力作用。 （-）多沙唑嗪和（）多沙唑嗪使去甲肾上腺素（主动脉）和去氧肾上腺素（前列腺平滑肌）的浓度-收缩曲线向右移动。大鼠主动脉中（-）恶唑烷的pA（2）值（8.625±0.053）小于（）恶唑烷（9.503±0.051），但它们在兔前列腺中的pK（B）值​​相同。在大鼠和兔子的心脏组织中，（）多沙唑嗪（3-30 µmol·L（-1））显着降低心房率，并产生负性变力作用；然而，（-）doxazosin不会影响心房率，并在心房产生正性肌力作用。因此，多沙唑嗪的手性碳原子不影响其对前列腺中α（1A）-肾上腺素受体治疗靶点的活性，但会显着改变其对主动脉中α（1D）-肾上腺素受体的阻断活性，并产生相反的正性肌力作用通过独立于α（1）-肾上腺素受体机制的心房。

BACKGROUND & AIMS: :Spontaneous intracranial hemorrhage is a debilitating form of stroke, often leading to death or permanent cognitive impairment. Many of the causative genes and the underlying mechanisms implicated in developmental cerebral-vascular malformations are unknown. Recent in vitro and in vivo studies in mice have shown inhibition of the 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) pathway to be effective in stabilizing cranial vessels. Using a combination of pharmacological and genetic approaches to specifically inhibit the HMGCR pathway in zebrafish (Danio rerio), we demonstrate a requirement for this metabolic pathway in developmental vascular stability. Here we report that inhibition of HMGCR function perturbs cerebral-vascular stability, resulting in progressive dilation of blood vessels, followed by vessel rupture, mimicking cerebral cavernous malformation (CCM)-like lesions in humans and murine models. The hemorrhages in the brain are rescued by prior exogenous supplementation with geranylgeranyl pyrophosphate (GGPP), a 20-carbon metabolite of the HMGCR pathway, required for the membrane localization and activation of Rho GTPases. Consistent with this observation, morpholino-induced depletion of the β-subunit of geranylgeranyltransferase I (GGTase I), an enzyme that facilitates the post-translational transfer of the GGPP moiety to the C-terminus of Rho family of GTPases, mimics the cerebral hemorrhaging induced by the pharmacological and genetic ablation of HMGCR. In embryos with cerebral hemorrhage, the endothelial-specific expression of cdc42, a Rho GTPase involved in the regulation of vascular permeability, was significantly reduced. Taken together, our data reveal a metabolic contribution to the stabilization of nascent cranial vessels, requiring protein geranylgeranylation acting downstream of the HMGCR pathway.

背景与目标： ：自发性颅内出血是中风的一种虚弱形式，通常会导致死亡或永久性认知障碍。涉及发展性脑血管畸形的许​​多致病基因和潜在机制尚不清楚。最近在小鼠中进行的体外和体内研究表明，抑制3-羟基-3-甲基戊二酰辅酶A还原酶（HMGCR）途径可有效稳定颅骨血管。使用药理和遗传学方法的组合来特异性抑制斑马鱼（Danio rerio）中的HMGCR途径，我们证明了这种代谢途径在发育性血管稳定性中的需求。在这里我们报道抑制HMGCR功能扰乱脑血管稳定性，导致血管进行性扩张，继而血管破裂，在人和鼠模型中模仿脑海绵状畸形（CCM）样病变。可以通过事先外源补充geranylgeranyl焦磷酸（GGPP）（HMGCR途径的20个碳代谢物）进行膜定位和激活Rho GTPases来挽救脑部出血。与该观察结果一致，吗啉代诱导的香叶基香叶基转移酶I（GGTase I）的β亚基耗竭，该酶有助于将GGPP部分翻译后转移至GTPases Rho家族的C末端，从而模拟脑出血由HMGCR的药理和遗传消融诱导。在患有脑出血的胚胎中，参与血管通透性调节的Rho GTPase cdc42的内皮特异性表达显着降低。综上所述，我们的数据揭示了新陈代谢对稳定新生颅血管的作用，需要在HMGCR途径下游作用的蛋白香叶基香叶基化作用。