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。

BACKGROUND & AIMS: :The advance of mitral valve repair techniques through tissue engineering is impeded by the lack of information regarding the cellular and extracellular components of the mitral valve. The present study aims to expand our understanding of the mitral valve structure by analysing the synthesis of extracellular matrix (ECM) proteins and the expression of nitric oxide synthase (NOS). Valvular endothelial cells (VECs) and valvular interstitial cells (VICs) were isolated from porcine mitral valves. Immunochemical staining of ECM components, including type I, II, III, IV and V collagen, laminin, fibronectin, elastin and chondroitin sulphate (CS), was performed on both mitral valve tissue and cell cultures. Reverse transcription polymerase chain reaction and immunochemistry were used to analyse NOS expression in native valve and in culture. Both VECs and VICs synthesised the basement membrane components, laminin and type IV collagen both in vivo and in vitro, amongst other fibrous ECM proteins. Synthesis of type I collagen and CS was absent in VEC cultures. Each cell type had a characteristic profile of NOS expression. VECs synthesised endothelial NOS both in vivo and in vitro, with a minority of VICs expressing neuronal NOS in vitro. The present study reports newly recognised aspects of the mitral valve structure and the in vitro behaviour of mitral valve cell populations based on ECM synthesis and NOS expression. The presented profiles can be used as base tools for the generation of data necessary for the selection of ideal cell sources and for the design of appropriate scaffolds for the development of effective tissue-engineered mitral valves.

背景与目标： : 缺乏有关二尖瓣细胞和细胞外成分的信息，阻碍了通过组织工程进行二尖瓣修复技术的发展。本研究旨在通过分析细胞外基质 (ECM) 蛋白的合成和一氧化氮合酶 (NOS) 的表达来扩大我们对二尖瓣结构的理解。从猪二尖瓣分离瓣膜内皮细胞 (VECs) 和瓣膜间质细胞 (VICs)。对二尖瓣组织和细胞培养物进行了ECM成分的免疫化学染色，包括I，II，III，IV和V型胶原蛋白，层粘连蛋白，纤连蛋白，弹性蛋白和硫酸软骨素 (CS)。逆转录聚合酶链反应和免疫化学用于分析天然瓣膜和培养物中NOS的表达。VECs和VICs均在体内和体外合成了基底膜成分，层粘连蛋白和IV型胶原蛋白以及其他纤维ECM蛋白。在VEC培养物中没有I型胶原蛋白和CS的合成。每种细胞类型都有NOS表达的特征。VECs在体内和体外合成了内皮NOS，少数VICs在体外表达神经元NOS。本研究报告了基于ECM合成和NOS表达的二尖瓣结构和二尖瓣细胞群体的体外行为的新认识。所呈现的配置文件可用作基础工具，用于生成选择理想细胞源所需的数据以及设计用于开发有效组织工程二尖瓣的适当支架。

BACKGROUND & AIMS: BACKGROUND:Trabecular bone strength diminishes as a result of osteoporosis and altered biomechanical loading at the vertebral and spinal levels. The spine consists of the anterior, middle and posterior columns and the load supported by the anterior and middle columns will differ across different regions of the spine. Stress shielding of the anterior column can contribute to bone loss and increase the risk of wedge fracture. There is a lack of quantitative data related to regional spinal bone mineral density distribution over time. We hypothesize that there is an increase in the posterior-to-anterior vertebral body bone mineral density ratio and a decrease in whole-body bone mineral density over time. METHODS:Bone mineral density was measured in 33 subjects using quantitative computed tomography scans for L1-L3 vertebrae, region (anterior and posterior vertebral body), and time (baseline and 6 years after). FINDINGS:Lumbar bone mineral density decreased significantly (Δ: ~15%) from baseline to the 6th year visit. Individual vertebra differences over time (L1: ~14%, L2: ~14%, L3: ~17%) showed statistical significance. Anterior bone mineral density change was significantly greater than in the posterior vertebral body region (Δ anterior: ~18%; Δ posterior: ~13%). Posterior-to-anterior bone mineral density ratio was significantly greater in the 6th year compared to baseline values (mean (SD), 1.33 (0.2) vs. 1.23 (0.1)). INTERPRETATION:This study provides longitudinal quantitative measurement of bone mineral density in vertebrae as well as regional changes in the anterior and posterior regions. Understanding bone mineral density distribution over time may help to decrease the risk of wedge fractures if interventions can be developed to bring spine loading to its normal state.

背景与目标：

BACKGROUND & AIMS: :Mitochondrial dysfunction is the most prominent source of oxidative stress in acute and chronic kidney disease. NLRX1 is a receptor of the innate immune system that is ubiquitously expressed and localized in mitochondria. We investigated whether NLRX1 may act at the interface of metabolism and innate immunity in a model of oxidative stress. Using a chimeric mouse model for renal ischemia-reperfusion injury, we found that NLRX1 protects against mortality, mitochondrial damage, and epithelial cell apoptosis in an oxidative stress-dependent fashion. We found that NLRX1 regulates oxidative phosphorylation and cell integrity, whereas loss of NLRX1 results in increased oxygen consumption, oxidative stress, and subsequently apoptosis in epithelial cells during ischemia-reperfusion injury. In line, we found that NLRX1 expression in human kidneys decreased during acute renal ischemic injury and acute cellular rejection. Although first implicated in immune regulation, we propose that NLRX1 function extends to the control of mitochondrial activity and prevention of oxidative stress and apoptosis in tissue injury.

背景与目标： : 线粒体功能障碍是急性和慢性肾脏疾病中氧化应激的最突出来源。NLRX1是先天免疫系统的受体，广泛表达并定位在线粒体中。我们研究了NLRX1是否可能在氧化应激模型中作用于代谢和先天免疫的界面。使用嵌合小鼠肾缺血再灌注损伤模型，我们发现NLRX1以氧化应激依赖性方式保护死亡率，线粒体损伤和上皮细胞凋亡。我们发现NLRX1调节氧化磷酸化和细胞完整性，而NLRX1的丢失会导致缺血再灌注损伤期间的耗氧量增加，氧化应激以及上皮细胞的凋亡。我们发现，在急性肾缺血损伤和急性细胞排斥反应期间，人肾脏中NLRX1的表达降低。尽管首先涉及免疫调节，但我们建议NLRX1功能扩展到控制线粒体活性以及预防组织损伤中的氧化应激和凋亡。

BACKGROUND & AIMS: :In a study of 55 patients with either acute lymphoid leukemia (ALL; 25 cases) or acute myeloid leukemia (AML; 30 cases), paraffin-embedded bone marrow particle sections were examined with a panel of monoclonal and polyclonal antibodies reactive toward lymphoid and myeloid-associated antigens, using the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique. All cases were previously classified according to the French-American-British (FAB) Co-operative Group, and cases of ALL were immunophenotyped by flow cytometry. Results indicated that myeloid-associated antibodies (Mac 387, KP 1 [CD68], antielastase, antilactoferrin, and antilysozyme) did not react with any case of ALL, M1-AML, or M6-AML, whereas at least one of these antibodies reacted with 20 of 21 (95%) cases of M2, M3, M4, and M5-AML. Anti-glycophorin C marked cases of M6-AML, whereas anti-CD3 labeled T-cell ALL. None of the antibodies tested specifically identified cases of B-cell ALL. The authors conclude that use of a selected panel of antibodies on paraffin-embedded bone marrow particle sections may be of value in the diagnosis and immunophenotypic classification of many cases of acute leukemias.

背景与目标： : 在对55例急性淋巴白血病 (ALL; 25例) 或急性髓细胞性白血病 (AML; 30例) 患者的研究中，用一组对淋巴样和髓样相关抗原具有反应性的单克隆和多克隆抗体检查了石蜡包埋的骨髓颗粒切片，使用碱性磷酸酶-抗碱性磷酸酶 (APAAP) 技术。所有病例先前均按法美英 (FAB) 合作组分类，所有病例均通过流式细胞术进行免疫表型。结果表明，髓样相关抗体 (Mac 387，KP 1 [CD68]，抗弹性蛋白酶，抗乳铁蛋白和抗溶菌酶) 未与ALL，M1-AML或M6-AML的任何病例发生反应，而这些抗体中的至少一种与21例 (95% 例) M2，M3，m4和M5-AML。抗糖蛋白C标记了M6-AML病例，而anti-CD3标记了T细胞ALL。测试的抗体均未特异性鉴定出b细胞ALL病例。作者得出的结论是，在石蜡包埋的骨髓颗粒切片上使用选定的一组抗体可能对许多急性白血病病例的诊断和免疫表型分类具有价值。

BACKGROUND & AIMS: BACKGROUND:Interstitial lung disease (ILD) frequently complicates systemic autoimmune disorders resulting in considerable morbidity and mortality. The connective tissue diseases (CTDs) most frequently resulting in ILD include: systemic sclerosis, idiopathic inflammatory myositis (including dermatomyositis, polymyositis and anti-synthetase syndrome) and mixed connective tissue disease. Despite the development, over the last two decades, of a range of biological therapies which have resulted in significant improvements in the treatment of the systemic manifestations of CTD, the management of CTD-associated ILD has changed little. At present there are no approved therapies for CTD-ILD. Following trials in scleroderma-ILD, cyclophosphamide is the accepted standard of care for individuals with severe or progressive CTD-related ILD. Observational studies have suggested that the anti-CD20 monoclonal antibody, rituximab, is an effective rescue therapy in the treatment of refractory CTD-ILD. However, before now, there have been no randomised controlled trials assessing the efficacy of rituximab in this treatment population. METHODS/DESIGN:RECITAL is a UK, multicentre, prospective, randomised, double-blind, double-dummy, controlled trial funded by the Efficacy and Mechanism Evaluation Programme of the Medical Research Council and National Institute for Health Research. The trial will compare rituximab 1 g given intravenously, twice at an interval of 2 weeks, with intravenously administered cyclophosphamide given monthly at a dose of 600 mg/m2 body surface area in individuals with ILD due to systemic sclerosis, idiopathic inflammatory myositis (including anti-synthetase syndrome) or mixed connective tissue disease. A total of 116 individuals will be randomised 1:1 to each of the two treatment arms, with stratification based on underlying CTD, and will be followed for a total of 48 weeks from first dose. The primary endpoint for the study will be change in forced vital capacity (FVC) at 24 weeks. Key secondary endpoints include: safety, change in FVC at 48 weeks as well as survival, change in oxygen requirements, total 48-week corticosteroid exposure and utilisation of health care resources. DISCUSSION:This is the first randomised control trial to study the efficacy of rituximab as first-line treatment in CTD-associated ILD. The results generated should provide important information on the treatment of a life-threatening complication affecting a rare group of CTDs. TRIAL REGISTRATION:ClinicalTrials.gov, NCT01862926. Registered on 22 May 2013.

背景与目标：

BACKGROUND & AIMS: PURPOSE:In the regulation of chromatin-structure and histone function, histone deacetylases (HDACs) are key enzymes and thus modulators of epigenetic regulation and gene expression. Accesses of the HDAC inhibitor vorinostat to intracellular compartments are essential to exert epigenetic effects. METHODS:In ten sarcoma patients receiving oral Zolinza (400 mg qd) vorinostat concentrations in plasma and peripheral blood mononuclear cells (PBMCs) were quantified using validated LC/MS/MS assays to determine intracellular and extracellular pharmacokinetic data. Cellular HDAC activity was evaluated using a fluorogenic assay. Concentration-response relationships were established between intracellular and extracellular vorinostat concentrations and HDAC inhibition in PBMCs. RESULTS:Pharmacokinetics of vorinostat and its two main inactive metabolites were determined over 8 h in plasma and PBMCs. Steady state AUCs (±SD) and T1/2 (±SD) were calculated to 4.61 ± 0.87 h µM and 1.73 ± 0.69 h (plasma) and 15.2 ± 9.03 h µM and 5.30 ± 4.27 h (PBMCs). Intracellular accumulation of vorinostat was determined together with prolonged vorinostat elimination in PBMCs. Cellular HDAC inhibition increased parallel with vorinostat concentrations in plasma and PBMCs. For effective inhibition of cellular HDACs (IC50) vorinostat concentrations of 0.05 µM in plasma and 0.17 µM in PBMCs were necessary. CONCLUSION:HDAC inhibition closely followed intracellular vorinostat concentrations and was short-lasting, which may contribute to the limited efficacy seen with vorinostat in solid tumors so far.

背景与目标：