BACKGROUND & AIMS: :Elucidation of the cellular basis of arrhythmias in ion channelopathy disorders is complicated by the inherent difficulties in studying human cardiac tissue. Thus we used a computer modeling approach to study the mechanisms of cellular dysfunction induced by mutations in inward rectifier potassium channel (K(ir))2.1 that cause Andersen-Tawil syndrome (ATS). ATS is an autosomal dominant disorder associated with ventricular arrhythmias that uncommonly degenerate into the lethal arrhythmia torsade de pointes. We simulated the cellular and tissue effects of a potent disease-causing mutation D71V K(ir)2.1 with mathematical models of human ventricular myocytes and a bidomain model of transmural conduction. The D71V K(ir)2.1 mutation caused significant action potential duration prolongation in subendocardial, midmyocardial, and subepicardial myocytes but did not significantly increase transmural dispersion of repolarization. Simulations of the D71V mutation at shorter cycle lengths induced stable action potential alternans in midmyocardial, but not subendocardial or subepicardial cells. The action potential alternans was manifested as an abbreviated QRS complex in the transmural ECG, the result of action potential propagation failure in the midmyocardial tissue. In addition, our simulations of D71V mutation recapitulate several key ECG features of ATS, including QT prolongation, T-wave flattening, and QRS widening. Thus our modeling approach faithfully recapitulates several features of ATS and provides a mechanistic explanation for the low frequency of torsade de pointes arrhythmia in ATS.

背景与目标： : 阐明离子通道病疾病中心律失常的细胞基础因研究人类心脏组织的固有困难而变得复杂。因此，我们使用计算机建模方法来研究由引起Andersen-Tawil综合征 (ATS) 的内向整流钾通道 (K(ir))2.1突变引起的细胞功能障碍的机制。ATS是一种常染色体显性遗传性疾病，与室性心律失常相关，通常退化为致死性心律失常。我们用人类心室肌细胞的数学模型和透壁传导的双齿瘤模型模拟了有效的致病突变D71V K(ir)2.1的细胞和组织效应。D71V K(ir)2.1突变导致心内膜下，中心肌和心外膜下肌细胞的动作电位持续时间显着延长，但并未显着增加复极的透壁离散度。在较短的周期长度上模拟D71V突变会在中段心肌中诱导稳定的动作电位交替，但不诱导心内膜下或心外膜下细胞。在透壁ECG中，动作电位交替体表现为缩写的QRS复合体，这是动作电位在中段心肌组织中传播失败的结果。此外，我们对D71V突变的模拟概括了ATS的几个关键ECG特征，包括QT延长，T波平坦化和QRS增宽。因此，我们的建模方法忠实地概括了ATS的几个特征，并为ATS中尖端扭转型心律失常的低频率提供了机械解释。

BACKGROUND & AIMS: The Dlx homeobox gene family is expressed in a complex pattern within the embryonic craniofacial ectoderm and ectomesenchyme. A previous study established that Dlx-2 is essential for development of proximal regions of the murine first and second branchial arches. Here we describe the craniofacial phenotype of mice with mutations in Dlx-1 and Dlx-1 and -2. The skeletal and soft tissue analyses of mice with Dlx-1 and Dlx-1 and -2 mutations provide additional evidence that the Dlx genes regulate proximodistal patterning of the branchial arches. This analysis also elucidates distinct and overlapping roles for Dlx-1 and Dlx-2 in craniofacial development. Furthermore, mice lacking both Dlx-1 and -2 have unique abnormalities, including the absence of maxillary molars. Dlx-1 and -2 are expressed in the proximal and distal first and second arches, yet only the proximal regions are abnormal. The nested expression patterns of Dlx-1, -2, -3, -5, and -6 provide evidence for a model that predicts the region-specific requirements for each gene. Finally, the Dlx-2 and Dlx-1 and -2 mutants have ectopic skull components that resemble bones and cartilages found in phylogenetically more primitive vertebrates.

背景与目标： Dlx同源盒基因家族在胚胎颅面外胚层和外胚间质中以复杂的模式表达。先前的研究表明，Dlx-2对于鼠第一和第二鳃弓近端区域的发育至关重要。在这里，我们描述了具有Dlx-1和-2突变的小鼠的颅面表型。对具有Dlx-1和-2突变的小鼠的骨骼和软组织分析提供了额外的证据，表明Dlx基因调节鳃弓的近端模式。此分析还阐明了Dlx-1和Dlx-2在颅面发育中的独特和重叠作用。此外，缺少Dlx-1和-2的小鼠具有独特的异常，包括缺少上颌磨牙。Dlx-1和-2在近侧和远侧第一和第二拱形中表达，但只有近侧区域是异常的。Dlx-1、-2、-3、-5和-6的嵌套表达模式为预测每个基因的区域特异性需求的模型提供了证据。最后，Dlx-2和Dlx-1和-2突变体具有异位的头骨成分，类似于在系统发育上更原始的脊椎动物中发现的骨骼和软骨。

BACKGROUND & AIMS: :Construction of tissue microarrays (TMAs) to efficiently characterize large sets of noninvasive epithelial lesions in the breast by immunohistochemistry is an appealing investigative approach, but presents technical challenges. We report methodologic studies performed to optimize methods for building TMAs from noninvasive breast tissues collected in a large case-control study of breast cancer. Using a manual arraying technique with 2.0-mm diameter needles, we constructed TMAs from specimens obtained from 32 women with breast cancer containing the following targets: (1) 28 terminal duct lobular units (TDLUs); (2) 28 ductal carcinomas in situ, and (3) 23 invasive carcinomas. Using careful target selection, we achieved representation of approximately 80% of noninvasive targets with sustained preservation through section 30 of the TMAs. Immunohistochemical staining of TDLU targets demonstrated positive staining for estrogen receptor (ER) in 30.8% of tubules and for progesterone receptor (PR) in 50.0%. To establish an efficient method to evaluate staining results in TDLUs, we created a categorical scoring system to approximate the percentage of tubules containing positive stained cells (<10%, 10% to 50%, >or=50%), and compared the results with those obtained by tubule counting. Comparison between the two methods demonstrated exact agreement for 70.8% of ER and 79.2% of PR stains without two-category discrepancies. ER/PR expression levels in multiple (up to 4) noninvasive targets of the same tissue type (TDLU or DCIS) from a single block showed good correlation. These data suggest that it is feasible to produce TMAs of noninvasive breast structures, albeit with careful selection of targets, and that immunostains of such cores may permit efficient immunohistochemical characterization of peritumoral tissues. Additional exploration of this approach is needed.

背景与目标： : 通过免疫组织化学构建组织微阵列 (tma) 以有效表征乳房中大量非侵入性上皮病变是一种吸引人的研究方法，但存在技术挑战。我们报告了方法学研究，以优化从大型乳腺癌病例对照研究中收集的非侵入性乳腺组织中构建TMAs的方法。使用具有2.0毫米直径针头的手动排列技术，我们从32名乳腺癌妇女获得的标本中构建了TMAs，这些标本包含以下目标 :( 1) 28个末端导管小叶单位 (TDLUs); (2) 28个原位导管癌和 (3) 23个浸润性癌。通过仔细的靶标选择，我们通过TMAs的第30节获得了约80% 的非侵入性靶标的持续保存。TDLU靶标的免疫组织化学染色显示肾小管30.8% 中的雌激素受体 (ER) 和50.0% 中的孕激素受体 (PR) 呈阳性染色。为了建立一种有效的方法来评估TDLUs中的染色结果，我们创建了一个分类评分系统来近似包含阳性染色细胞 (<10%，10% 至50%，> 或 = 50%) 的小管的百分比，并将结果与通过小管计数获得的结果进行比较。两种方法之间的比较表明，ER的70.8% 和PR染色的79.2% 完全一致，没有两类差异。来自单个块的相同组织类型 (TDLU或DCIS) 的多个 (最多4个) 非侵入性靶标中的ER/PR表达水平显示出良好的相关性。这些数据表明，尽管仔细选择了靶标，但产生非侵入性乳房结构的tma是可行的，并且此类核心的免疫染色可能允许对瘤周组织进行有效的免疫组织化学表征。需要对这种方法进行进一步的探索。

BACKGROUND & AIMS: :A diagnosis of low-grade fibromyxoid sarcoma (LGFMS) remains problematic because of its bland-looking histologic features that can be potentially confused with other benign or low-grade fibromyxoid lesions. Recent cytogenetic and molecular analyses have shown that most LGFMSs have a characteristic chromosomal abnormality, t(7;16)(q33;p11), resulting in the FUS-CREB3L2 fusion gene. However, such assays have only rarely been used to analyze formalin-fixed, paraffin-embedded tumor samples. In the present study, we conducted a reverse transcription-polymerase chain reaction assay to detect the FUS-CREB3L2 fusion transcripts using formalin-fixed, paraffin-embedded tumor tissue specimens from 16 LGFMSs including 3 cases with giant collagen rosettes. The primers were newly designed to specifically amplify most of the junctional regions of the FUS-CREB3L2 fusion gene transcripts previously reported. The FUS-CREB3L2 fusion gene transcripts were detected in 14/16 (88%) cases of LGFMS. A nucleotide sequence analysis of the PCR products revealed that different portions of the FUS exon 6 or 7 were fused with various sites of the CREB3L2 exon 5, resulting in 12 different nucleotide sequences. We also tested a primer set to detect the FUS-CREB3L1 fusion transcript, which is a rare variant of the gene fusion in LGFMS, although no PCR products were identified in any case. The FUS-CREB3L2 fusion transcripts were not detected in any of the 123 other soft-tissue tumors, including desmoid-type fibromatoses, myxofibrosarcomas, soft-tissue perineuriomas, and congenital or adult fibrosarcomas. These data suggest that our reverse transcription-polymerase chain reaction assay is a reliable method to detect FUS-CREB3L2, which can thus help in accurately diagnosing LGFMS.

背景与目标： : 低度纤维粘液样肉瘤 (LGFMS) 的诊断仍然存在问题，因为其外观平淡无奇的组织学特征可能与其他良性或低度纤维粘液样病变相混淆。最近的细胞遗传学和分子分析表明，大多数lgfms具有特征性的染色体异常t(7;16)(q33;p11)，导致FUS-CREB3L2融合基因。然而，这种测定很少用于分析福尔马林固定的石蜡包埋的肿瘤样品。在本研究中，我们进行了逆转录聚合酶链反应分析，以使用来自16个lgfms的福尔马林固定，石蜡包埋的肿瘤组织标本 (包括3例巨大的胶原玫瑰花结) 检测FUS-CREB3L2融合转录本。新设计了引物，以特异性扩增先前报道的FUS-CREB3L2融合基因转录物的大部分连接区域。在14/16 (88%) 例LGFMS中检测到FUS-CREB3L2融合基因转录本。PCR产物的核苷酸序列分析表明，FUS外显子6或7的不同部分与CREB3L2外显子5的不同位点融合，产生12种不同的核苷酸序列。我们还测试了引物集以检测FUS-CREB3L1融合转录本，这是LGFMS中基因融合的罕见变体，尽管在任何情况下都没有鉴定出PCR产物。在123种其他软组织肿瘤中均未检测到FUS-CREB3L2的融合转录物，包括真丝样纤维瘤，粘液纤维肉瘤，软组织神经瘤和先天性或成人纤维肉瘤。这些数据表明，我们的逆转录-聚合酶链反应测定法是检测FUS-CREB3L2的可靠方法，因此可以帮助准确诊断LGFMS。

BACKGROUND & AIMS: BACKGROUND:Although the benefit of thrombolytic therapy in reducing mortality in acute myocardial infarction is well established, the types of bleeding and risk factors for bleeding are less well described in large trials.

METHODS AND RESULTS:We analyzed the baseline characteristics, outcomes, and incidence of bleeding by location, severity, and treatment assignment among 41,021 patients in the GUSTO-I trial of thrombolysis for acute myocardial infarction. Of the 40,903 patients for whom there were complete data, 1.2% suffered severe bleeding and 11.4% experienced moderate hemorrhage at a variety of sites. The most common sources of bleeding were procedure related. The thrombolytic regimen was strongly related to the incidence of bleeding; comparatively more bleeding was seen with the therapies of streptokinase plus intravenous heparin and the streptokinase and tissue plasminogen activator plus intravenous heparin combination. In multivariate analysis, the four most powerful independent predictors of hemorrhage were older age, lighter body weight, female sex, and African ancestry; they remained the most important predictors of bleeding when multivariate analysis was performed on patients who did not undergo invasive procedures. The presence of serious hemorrhage was associated with other undesirable outcomes (recurrent events, left ventricular dysfunction, arrhythmia, or stroke).

CONCLUSIONS:Important predictors of bleeding in this population are increased age, lighter weight, female sex, African ancestry, and experiencing invasive procedures. Other nonhemorrhagic adverse clinical outcomes were associated with moderate and severe bleeding, which was in turn associated with increased length of hospital stay and mortality at 30 days.

背景与目标： 背景 : 尽管溶栓治疗在降低急性心肌梗死死亡率方面的益处已得到充分证实，但在大型试验中，出血类型和出血危险因素的描述较少。
方法和结果 : 在GUSTO-I急性心肌梗死溶栓试验中，我们分析了41,021名患者的基线特征，结局和出血发生率，按位置，严重程度和治疗分配。在40,903例有完整数据的患者中，1.2% 例严重出血，11.4% 例在不同部位出现中度出血。最常见的出血来源与手术有关。溶栓方案与出血发生率密切相关; 链激酶加静脉肝素以及链激酶和组织型纤溶酶原激活剂加静脉肝素联合治疗的出血相对较多。在多变量分析中，出血的四个最有效的独立预测因子是年龄较大，体重较轻，女性和非洲血统; 当对未接受侵入性操作的患者进行多变量分析时，它们仍然是出血的最重要预测因子。严重出血的存在与其他不良结果 (复发事件，左心室功能障碍，心律失常或中风) 相关。
结论 : 该人群出血的重要预测因素是年龄增加，体重减轻，女性，非洲血统，经历侵入性手术。其他非出血性不良临床结局与中度和重度出血相关，而中度和重度出血又与30天时的住院时间和死亡率增加相关。

BACKGROUND & AIMS: BACKGROUND AND AIMS OF THE STUDY:Bovine and porcine pericardial tissues stabilized by dye-mediated photooxidation have found application as bioprosthetic heart valve material.

METHODS:To help predict clinical performance, a series of tests were performed to assess the biocompatibility and immunologic properties of these materials.

RESULTS AND CONCLUSIONS:Photooxidized bovine or porcine pericardium sterilized with an iodine-based solution were found to be non-cytotoxic, non-hemolytic, and non-mutagenic. Oil or saline extracts of these tissues passed tests for intracutaneous toxicity (irritation), acute systemic toxicity, and subchronic toxicity. Histopathology of 90-day implants of these tissues in the rabbit model demonstrated no significant macroscopic reaction and only slight microscopic response. Using a rabbit model to assess immune response, both bovine and porcine pericardial tissues elicited low levels of antibody. Furthermore, tissue photooxidation or iodine sterilization did not increase the overall level of antibodies. Glutaraldehyde-treated tissue also elicited low antibody levels which were higher than photooxidized tissue-induced levels. Absorption studies indicated that the photooxidation process may generate new epitopes, possibly collagen cross-links. Using the juvenile sheep model to assess in vivo performance, bioprosthetic valves made with photooxidized tissue were implanted and allowed to serve as functional implants for up to two years. Upon explant, the photooxidized pericardial leaflets were found to be non-calcific and partially covered with a layer of host cells. Histological cross-sections stained with von Willebrand's factor confirmed this layer as endothelial cells.

背景与目标： 研究的背景和目的 : 通过染料介导的光氧化作用稳定的牛和猪心包组织已被用作生物人工心脏瓣膜材料。
方法 : 为了帮助预测临床表现，进行了一系列测试以评估这些材料的生物相容性和免疫学特性。
结果和结论 : 发现用碘基溶液灭菌的光氧化牛或猪心包具有非细胞毒性，非溶血性，和非诱变性。这些组织的油或盐水提取物通过了皮内毒性 (刺激)，急性全身毒性和亚慢性毒性的测试。在兔模型中，这些组织的90天植入物的组织病理学没有明显的宏观反应，只有轻微的微观反应。使用兔模型评估免疫反应，牛和猪心包组织均引起低水平的抗体。此外，组织光氧化或碘灭菌不会增加抗体的总体水平。戊二醛处理的组织也引起低抗体水平，该水平高于光氧化组织诱导的水平。吸收研究表明，光氧化过程可能会产生新的表位，可能是胶原蛋白交联。使用幼年绵羊模型评估体内性能，将用光氧化组织制成的生物人工瓣膜植入并作为功能性植入物长达两年。外植体后，发现光氧化的心包小叶是非钙化的，并部分覆盖有一层宿主细胞。用von Willebrand因子染色的组织学横截面证实了该层为内皮细胞。

BACKGROUND & AIMS: :The distributed genome hypothesis (DGH) states that each strain within a bacterial species receives a unique distribution of genes from a population-based supragenome that is many times larger than the genome of any given strain. The observations that natural infecting populations are often polyclonal and that most chronic bacterial pathogens have highly developed mechanisms for horizontal gene transfer suggested the DGH and provided the means and the mechanisms to explain how chronic infections persist in the face of a mammalian host's adaptive defense mechanisms. Having previously established the validity of the DGH for obligate pathogens, we wished to evaluate its applicability to an opportunistic bacterial pathogen. This was accomplished by construction and analysis of a highly redundant pooled genomic library containing approximately 216,000 functional clones that was constructed from 12 low-passage clinical isolates of Pseudomonas aeruginosa, 6 otorrheic isolates and 6 from other body sites. Sequence analysis of 3,214 randomly picked clones (mean insert size, approximately 1.4 kb) from this library demonstrated that 348 (10.8%) of the clones were unique with respect to all genomic sequences of the P. aeruginosa prototype strain, PAO1. Hypothetical translations of the open reading frames within these unique sequences demonstrated protein homologies to a number of bacterial virulence factors and other proteins not previously identified in P. aeruginosa. PCR and reverse transcription-PCR-based assays were performed to analyze the distribution and expression patterns of a 70-open reading frame subset of these sequences among 11 of the clinical strains. These sequences were unevenly distributed among the clinical isolates, with nearly half (34/70) of the novel sequences being present in only one or two of the individual strains. Expression profiling revealed that a vast majority of these sequences are expressed, strongly suggesting they encode functional proteins.

背景与目标： : 分布式基因组假说 (DGH) 指出，细菌物种中的每个菌株都从基于群体的上基因组接收到独特的基因分布，该基因分布比任何给定菌株的基因组大很多倍。观察到自然感染种群通常是多克隆的，并且大多数慢性细菌病原体具有高度发达的水平基因转移机制，这表明DGH并提供了手段和机制来解释慢性感染如何在哺乳动物宿主的适应性防御机制中持续存在。先前已经确定了DGH对专性病原体的有效性，我们希望评估其对机会性细菌病原体的适用性。这是通过构建和分析包含约216,000个功能克隆的高度冗余的合并基因组文库来实现的，所述基因组文库是从铜绿假单胞菌的12个低传代临床分离株、6个耳鼻喉科分离株和6个来自其他身体位点构建的。对从该文库中随机挑选的3,214个克隆 (平均插入大小，约1.4 kb) 的序列分析表明，对于铜绿假单胞菌原型菌株pao1的所有基因组序列，348 (10.8%) 的克隆是独特的。这些独特序列中开放阅读框的假设翻译证明了蛋白质与许多细菌毒力因子和其他以前在铜绿假单胞菌中未鉴定的蛋白质的同源性。进行了PCR和基于逆转录PCR的检测，以分析11种临床菌株中这些序列的70个开放阅读框子集的分布和表达模式。这些序列在临床分离株中分布不均匀，近一半 (34/70) 新序列仅存在于单个菌株中的一个或两个中。表达谱分析显示，这些序列中的绝大多数都被表达，这强烈表明它们编码功能蛋白。

BACKGROUND & AIMS: BACKGROUND:A novel classification of alpha-1 adrenoceptor subtypes (High, Low) was applied to human benign prostatic hypertrophy (BPH) tissue. METHODS:Human BPH specimens were examined by a radioligand binding assay method using 3H-prazosin, and those data were compared with preoperative therapies. RESULTS:(1) Scatchard analysis showed a high-affinity site (Kd:27.18 +/- 6.41 pM; Bmax:9.29 +/- 0.98 fM/mg protein; mean +/- SE) as alpha 1H, and a low-affinity site (Kd: 4088.0 +/- 744.34 pM, Bmax: 140.81 +/- 19.98 fM/mg protein) as alpha 1L subtype, for prazosin. (2) The Kd and Bmax were not different in the nontreated group (n = 5), alpha 1 blocker group (n = 5), and antiandrogen group (n = 5), in either alpha 1-high affinity or alpha 1-low affinity subtype. (3) Phenoxybenzamine had different pKi values for the above two adrenoceptor subtypes. Scatchard analysis showed that alpha 1-high affinity binding site disappeared in the presence of 1 microM of phenoxybenzamine, and the Kd and Bmax values in the presence of 1 microM of phenoxybenzamine were almost identical to the alpha 1-low affinity site of the two subtypes. CONCLUSIONS:Human BPH tissue possesses both alpha 1H- and alpha 1L-adrenoceptor subtypes according to the affinities for prazosin, and only the alpha 1H subtype can be completely inhibited by some concentration of phenoxybenzamine. Treatment by alpha 1 blocker may not change the conditions of alpha 1-adrenoceptors in prostatic tissue.

背景与目标：

BACKGROUND & AIMS: :The distribution of calcitonin gene-related peptide (CGRP), enkephalin, galanin, neuropeptide Y (NPY), somatostatin, tachykinins and vasoactive intestinal polypeptide (VIP) was compared in cervical, thoracic, lumbar and sacral segmental levels of spinal cord and dorsal root ganglia of horse and pig. In both species, immunoreactivity for the peptides under study was observed at all segmental levels of the spinal cord. Peptide-immunoreactive fibres were generally concentrated in laminae I-III, the region around the central canal, and in the autonomic nuclei. A general increase in the number of immunoreactive nerve fibres was noted in the lumbosacral segments of the spinal cord, which was particularly exaggerated in the case of VIP immunoreactivity. In the horse, some CGRP-, somatostatin- or tachykinin-immunoreactive cell bodies were present in the dorsal horn. In the pig, cells immunoreactive for somatostatin, enkephalin or NPY were noted in a similar location. In the ventral horn most motoneurones were CGRP-immunoreactive in both species. However, in pig many other cell types were CGRP-immunoreactive not only in the ventral horn, but also in laminae V-VI of the dorsal horn. With the exception of enkephalin and NPY immunoreactivity, which was not seen in pig dorsal root ganglia, all peptides studied were localised to neuronal cell bodies and/or fibres in the dorsal root ganglia. In both species, immunolabeled cell bodies were observed in ganglia from cervical, thoracic, lumbar and sacral levels, with the exception of VIP-immunoreactive cells that were detected only in the lumbosacral ganglia. Numerous CGRP- and tachykinin-immunoreactive cell bodies were visualised in both species, while the cells immunolabeled with other peptide antisera were much lower in number. In both species, immunostaining of serial sections revealed that a subset of CGRP-immunoreactive cells co-expressed tachykinin, galanin or somatostatin immunoreactivity. In the horse some enkephalin-immunoreactive cells were also CGRP positive and occasionally combinations of three peptides, e.g. CGRP, tachykinin and galanin or CGRP, tachykinin and enkephalin were identified. The results obtained suggest that the overall pattern of distribution of peptide immunoreactivities is in general agreement with that so far described in other mammals, although some species variations have been observed, particularly regarding the presence of immunoreactive cell bodies in the dorsal horn of the spinal cord.

背景与目标： 比较了马、猪颈、胸、腰、骶段脊髓和背根神经节中降钙素基因相关肽 (CGRP) 、脑啡肽、甘丙肽、神经肽Y (NPY) 、生长抑素、速激肽和血管活性肠多肽 (VIP) 的分布。在这两个物种中，在脊髓的所有节段水平上都观察到了所研究肽的免疫反应性。肽免疫反应性纤维通常集中在laminae i-iii，中央管周围的区域和自主核中。在脊髓的腰s节段中，免疫反应性神经纤维的数量普遍增加，在VIP免疫反应性的情况下，这一点尤其严重。在马中，背角中存在一些CGRP，生长抑素或速激肽免疫反应性细胞体。在猪中，对生长抑素，脑啡肽或NPY具有免疫反应性的细胞位于相似的位置。在腹侧角中，大多数运动神经元在两种物种中均具有CGRP免疫反应性。然而，在猪中，许多其他类型的细胞不仅在腹角而且在背角的层v-vi中都具有CGRP免疫反应性。除了在猪背根神经节中未发现的脑啡肽和NPY免疫反应性外，所有研究的肽均定位于背根神经节中的神经元细胞体和/或纤维。在这两个物种中，在神经节中从颈，胸，腰和骶骨水平观察到免疫标记的细胞体，但仅在腰s神经节中检测到的VIP免疫反应性细胞除外。在这两个物种中均可见许多CGRP和速激肽免疫反应性细胞体，而用其他肽抗血清免疫标记的细胞数量要低得多。在这两个物种中，连续切片的免疫染色显示CGRP免疫反应性细胞的一部分共同表达速激肽，甘丙肽或生长抑素免疫反应性。在马中，一些脑啡肽免疫反应性细胞也是CGRP阳性，偶尔会结合三种肽，例如CGRP，速激肽和甘丙肽或CGRP，速激肽和脑啡肽被鉴定。获得的结果表明，肽免疫反应性的总体分布模式与迄今为止在其他哺乳动物中描述的基本一致，尽管已经观察到一些物种变化，特别是关于脊髓背角中存在免疫反应性细胞体。

BACKGROUND & AIMS: :Plectin is a widely expressed protein that is very large in size and that has all the attributes of a multifunctional crosslinking and organizing element of the cytoskeleton. It displays a multidomain structure, versatile binding activities, and subcellular localizations that enable it to strengthen cells against mechanical stress forces. Moreover, hereditary gene defects in plectin cause epidermolysis bullosa simplex (EBS)-MD, a severe skin blistering disease with muscular dystrophy. Here we report the analysis of the exonintron organization of the rat plectin gene and the identification of several different isoforms on the transcriptional level. We show that of 35 coding exons identified, 4 serve as alternative first exons splicing into the same successive exon 2, which is the first of 7 exons encoding a highly conserved actin-binding domain. RNase protection mapping of transcripts containing 3 of the identified 4 alternate first exons revealed their coexpression in rat glioma C6 cells and in a series of different rat tissues that we examined. Significant variations in expression levels of first exons indicated the possibility of tissue-specific promoter usage. In addition, plectin splice variants lacking exon 31 (> 3 kb), which encodes the entire rod domain of the molecule, were identified in a variety of rat tissues. This study provides first insights into a complex plectin gene regulatory machinery with similarities to that of dystrophin.

背景与目标： : pletin是一种广泛表达的蛋白质，其大小非常大，并且具有细胞骨架的多功能交联和组织元件的所有属性。它显示出多域结构，多种结合活性和亚细胞定位，使其能够增强细胞抵抗机械应力的能力。此外，plectin的遗传性基因缺陷会导致大疱性表皮松解症 (EBS)-MD，这是一种伴有肌营养不良的严重皮肤水疱性疾病。在这里，我们报告了对大鼠plectin基因外显子组织的分析，并在转录水平上鉴定了几种不同的同工型。我们显示，在鉴定出的35个编码外显子中，有4个作为替代的第一个外显子拼接成相同的连续外显子2，这是编码高度保守的肌动蛋白结合域的7个外显子中的第一个。包含鉴定出的4个交替第一外显子中的3个的转录物的RNase保护作图显示了它们在大鼠神经胶质瘤C6细胞和我们检查的一系列不同大鼠组织中的共表达。第一个外显子表达水平的显着变化表明使用组织特异性启动子的可能性。此外，在多种大鼠组织中鉴定出缺少外显子31 (> 3 kb) 的pletin剪接变体，该外显子编码分子的整个杆结构域。这项研究提供了与肌营养不良蛋白相似的复杂pletin基因调控机制的第一个见解。

BACKGROUND & AIMS: :An immediately inducible gene by gonadotropin was isolated from rat ovaries primed with pregnant mare serum gonadotropin (PMSG) by using a subtraction cloning procedure. Homology analysis revealed that the gene is a rat homologue of scavenger receptor class B-I, which was recently identified as a specific receptor for high density lipoprotein (HDL). The structure of rat SRBI was determined by nucleotide sequence analysis of full-length cDNAs for SRBI. Northern blot analysis revealed that rat SRBI mRNA levels were rapidly and strongly increased within 3 h by the injection of PMSG. In situ hybridization study revealed that SRBI mRNA was strongly induced in theca interna cells of immature rat ovary stimulated with 30 IU of PMSG for 6 h. SRBI mRNA expression was also observed in corpora lutea of the adult rat ovary. These findings indicate that expression of SRBI mRNA is restricted to and induced in the ovarian steroidogenic cell types where cholesterol is used as a substrate for synthesis of steroid hormones. Our data strongly suggest that SRBI may play a significant role in the ovarian steroidogenesis by mediating selective uptake of cholesterol from HDL to ovarian theca interna cells or to corpus luteum.

背景与目标： : 使用减法克隆程序从用妊娠母马血清促性腺激素 (PMSG) 引发的大鼠卵巢中分离出促性腺激素立即诱导的基因。同源性分析表明，该基因是清道夫受体B-I类的大鼠同源物，最近被鉴定为高密度脂蛋白 (HDL) 的特异性受体。通过SRBI全长cdna的核苷酸序列分析确定大鼠SRBI的结构。Northern印迹分析显示，通过注射PMSG，大鼠SRBI mRNA水平在3小时内迅速且强烈地增加。原位杂交研究表明，用30 IU的PMSG刺激6 h，在未成熟大鼠卵巢的卵泡膜细胞中强烈诱导SRBI mRNA。在成年大鼠卵巢的黄体中也观察到了SRBI mRNA的表达。这些发现表明，SRBI mRNA的表达仅限于并在其中胆固醇用作合成类固醇激素的底物的卵巢类固醇生成细胞类型中诱导。我们的数据强烈表明，SRBI可能通过介导胆固醇从HDL到卵巢卵泡膜细胞或黄体的选择性摄取而在卵巢类固醇生成中起重要作用。

BACKGROUND & AIMS: Mayaro virus (alphavirus) infection of Aedes albopictus cells results in inhibition of cell protein synthesis and viral proteins are preferably synthesized. When infected cells are heat shocked, however, there is also an inhibition of viral protein synthesis, and there is preferential synthesis of heat shock proteins. Based on these observations, the distribution of Mayaro viral RNA in polysomes and the association of p34 (capsid protein) with ribosomal fractions of the cells under such conditions have been analyzed. During infection, the viral RNA is mainly observed in light polysomes (60% of total viral RNA in the cell) and also in heavy polysomes (13%). However, when infected cells are heat-shocked, the viral RNA is strongly mobilized from heavy polysomes to the light polysomes fraction and an enrichment in the unbound fraction can be noticed. The amount of p34 associated with the ribosomal fraction was also shown to be decreased in the heat shocked cells. These data lead to the suggestion that two mechanisms could be involved in the inhibition of Mayaro virus protein synthesis in response to heat shock(1) mobilization of Mayaro virus RNA from heavy to light polysomes; (2) a decrease in the amount of the p34 within the ribosomal fraction.

背景与目标： 白纹伊蚊细胞的Mayaro病毒 (α 病毒) 感染导致细胞蛋白合成的抑制，优选合成病毒蛋白。然而，当受感染的细胞被热休克时，病毒蛋白的合成也受到抑制，并且热休克蛋白的优先合成。基于这些观察结果，已经分析了在这种条件下，多核糖体中Mayaro病毒RNA的分布以及p34 (衣壳蛋白) 与细胞核糖体部分的关联。在感染期间，病毒RNA主要在轻多核糖体 (60% 细胞中总病毒RNA) 和重多核糖体 (13%) 中观察到。但是，当受感染的细胞受到热冲击时，病毒RNA会从重多核糖体强烈动员到轻多核糖体部分，并且可以注意到未结合部分的富集。在热休克细胞中，与核糖体级分相关的p34的量也显示出减少。这些数据表明，两种机制可能参与抑制马亚罗病毒蛋白合成，以响应热休克 (1) 马亚罗病毒RNA从重到轻多核糖体的动员; (2) 核糖体部分中p34的含量减少。

BACKGROUND & AIMS: :CD3(-)CD4(+)CD45(+) inducer cells are required for the initiation of mucosa-associated organogenesis of both nasopharynx-associated lymphoid tissues (NALT) and Peyer's patches (PP) in the aerodigestive tract. CXCL13(-/-) mice and mice carrying the paucity of lymph node T cell (plt) mutation and lacking expression of CCL19 and CCL21 accumulate CD3(-)CD4(+)CD45(+) cells at the site of NALT but not of PP genesis. Although NALT was observed to develop in adult CXCL13(-/-) and plt/plt mice, the formation of germinal centers in CXCL13(-/-) mice was affected, and their population of B cells was much lower than in the NALT of CXCL13(+/-) mice. Similarly, fewer T cells were observed in the NALT of plt/plt mice than in control mice. These findings indicate that the initiation of NALT organogenesis is independent of CXCL13, CCL19, and CCL21. However, the expression of these lymphoid chemokines is essential for the maturation of NALT microarchitecture.

背景与目标： : CD3(-)CD4 () CD45 () 诱导细胞是启动鼻咽相关淋巴组织 (NALT) 和Peyer斑块 (PP) 的粘膜相关器官发生所必需的。CXCL13(-/-) 小鼠和携带缺乏淋巴结T细胞 (plt) 突变且缺乏CCL19和CCL21表达的小鼠在NALT位点积累CD3(-)CD4 () CD45 () 细胞，而不是PP发生。尽管在成年CXCL13(-/-) 和plt/plt小鼠中观察到NALT的发展，但CXCL13(-/-) 小鼠的生发中心的形成受到影响，其b细胞种群远低于CXCL13 (/-) 小鼠的NALT。同样，在plt/plt小鼠的NALT中观察到的T细胞少于对照小鼠。这些发现表明，NALT器官发生的启动与CXCL13，CCL19和ccl21无关。然而，这些淋巴趋化因子的表达对于NALT微结构的成熟至关重要。

BACKGROUND & AIMS: Although asthmatic patients are known to have increased levels of IgG antibody against house dust mite (HDM), it is not clear whether or not the presence of HDM-specific IgG antibody is associated with the etiological mechanism of asthma. To address this problem, we evaluated the relationship between HDM-specific IgG antibody levels and incidence of asthma in a general pediatric population. IgE and IgG antibody levels against Dermatophagoides farinae (Df) were examined by RAST and ELISA in a total of 722 randomly selected schoolchildren including 26 subjects with asthma, and the relative prevalence rates of asthma in this population were evaluated in relation to both Df-specific IgE and IgG levels. The incidence of asthma correlated not only with levels of Df-specific IgE, but also with those of Df-specific IgG. There was a significant correlation between Df-specific IgE and IgG levels both in the total population and in the asthmatic children. Because IgG and IgE responses occurred in parallel in this population, the clinical significance of HDM-specific IgG anti-body remains unclear. However, our findings have suggested that clinical expression of asthma in children is primarily dependent on their capacity to mount a immune response to HDM, which includes both IgE and IgG responses.

背景与目标： 尽管已知哮喘患者针对屋尘螨 (HDM) 的IgG抗体水平升高，但尚不清楚HDM特异性IgG抗体的存在是否与哮喘的病因机制有关。为了解决这个问题，我们评估了普通儿童人群中HDM特异性IgG抗体水平与哮喘发生率之间的关系。通过RAST和ELISA检测了总共722名随机选择的学童 (包括26名哮喘受试者) 的针对粉尘螨 (Df) 的IgE和IgG抗体水平，并评估了该人群中哮喘的相对患病率与Df特异性IgE和IgG水平。哮喘的发病率不仅与Df特异性IgE的水平相关，而且与Df特异性IgG的水平相关。在总人口和哮喘儿童中，Df特异性IgE和IgG水平之间存在显着相关性。由于IgG和IgE反应在该人群中并行发生，因此HDM特异性IgG抗体的临床意义尚不清楚。但是，我们的发现表明，儿童哮喘的临床表达主要取决于他们对HDM产生免疫反应的能力，其中包括IgE和IgG反应。

BACKGROUND & AIMS: :Isolated rat adipose tissue-derived stromal cells (rATSCs) contain pluripotent cells that can be differentiated into a variety of cell lineages, including neural cells. Recent work has shown that ATSCs can make neurosphere-like clumps and differentiate into neuron-like cells expressing neuronal markers, but their therapeutic effect is unclear. Here we report that intravenous infusion of oligodendrocyte precursor cells (OPCs) derived from rATSC autograft cells sources improve motor function in rat models of spinal cord injury (SCI). After 4-5 weeks, transplanted rATSC-OPC cells survived and migrated into the injured region of SCI very efficiently (30-35%) and migrated cells were partially differentiated into neurons and oligodendrocyte. Also, we found some of the engrafted OPCs migrated and integrated in the kidney, brain, lung, and liver through the intravenous system. Behavioral analysis revealed the locomotor functions of OPC-autografted SCI rats were significantly restored. Efficient migration of intravenously engrafted rATSC-OPCs cells into SCI lesion suggests that SCI-induced chemotaxic factors facilitate migration of rATSC-OPCs. Here, we verified that engrafted rATSCs and SCI-induced chemotaxic factors indeed play an important role in proliferation, migration, and differentiation of endogeneous spinal cord-derived neural progenitor cells in the injured region. In transplantation paradigms, the interaction between engrafted rATSC-OPCs and endogeneous spinal cord-derived neuronal progenitor cells will be important in promoting healing through fate decisions, resulting in coordinated induction of cell migration and differentiation.

背景与目标： : 分离的大鼠脂肪组织来源的基质细胞 (rATSCs) 含有多能细胞，可分化为多种细胞谱系，包括神经细胞。最近的研究表明，ATSCs可以形成神经球状团块并分化为表达神经元标记的神经元样细胞，但其治疗效果尚不清楚。在这里，我们报告了来自rATSC自体移植细胞来源的少突胶质前体细胞 (opc) 的静脉输注改善了大鼠脊髓损伤 (SCI) 模型的运动功能。4-5周后，移植的ratsc-opc细胞存活并非常有效地迁移到SCI的损伤区域 (30-35%)，并且迁移的细胞部分分化为神经元和少突胶质细胞。此外，我们发现一些移植的opc通过静脉系统迁移并整合到肾脏，大脑，肺和肝脏中。行为分析表明，OPC自体移植的SCI大鼠的运动功能显着恢复。静脉移植的ratsc-opcs细胞有效迁移到SCI病变中，表明SCI诱导的趋化因子促进了ratsc-opcs的迁移。在这里，我们证实了移植的大鼠干细胞和SCI诱导的趋化因子确实在受损区域内源性脊髓神经祖细胞的增殖，迁移和分化中起重要作用。在移植范例中，移植的ratsc-opcs与内源性脊髓衍生的神经元祖细胞之间的相互作用对于通过命运决定促进愈合非常重要，从而导致细胞迁移和分化的协调诱导。