BACKGROUND & AIMS: The involvement of antigen-specific T cells in the pathogenesis of collagen diseases is still controversial. The final stages of collagen diseases are usually characterized by the dominance of inflammation. Therefore, antigen non-specific factors, such as inflammatory cytokines, probably play an important role in this process. On the other hand, the methods available to analyze the antigen-specific aspects of the immune response are still limited. Here we review our novel system of T cell clonality analysis based on the idea that activated antigen-specific T cells should form accumulating clones among the lymphocyte population. Using this method, dynamic changes of clonal accumulation of T cells could be evaluated during antigenic stimulation in vivo and in vitro. The significance of antigen-specific T cell clones in collagen diseases is discussed using data obtained from patients with rheumatoid arthritis and systemic lupus erythematosus.

背景与目标： 抗原特异性T细胞是否参与胶原蛋白疾病的发病机制仍存在争议。胶原蛋白疾病的最后阶段通常以炎症占优势为特征。因此，抗原非特异性因子，例如炎性细胞因子，可能在此过程中起重要作用。另一方面，可用于分析免疫应答的抗原特异性方面的方法仍然有限。在这里，我们基于激活的抗原特异性T细胞应在淋巴细胞群体中形成累积克隆的想法，回顾了我们的T细胞克隆性分析的新系统。使用这种方法，可以在体内和体外抗原刺激过程中评估T细胞克隆积累的动态变化。利用类风湿性关节炎和系统性红斑狼疮患者获得的数据，讨论了抗原特异性T细胞克隆在胶原蛋白疾病中的重要性。

BACKGROUND & AIMS: :The transcription factor signal transducer and activator of transcription-1 (STAT1) plays a key role in immunity against mycobacterial and viral infections. Here, we characterize three human STAT1 germline alleles from otherwise healthy patients with mycobacterial disease. The previously reported L706S, like the novel Q463H and E320Q alleles, are intrinsically deleterious for both interferon gamma (IFNG)-induced gamma-activating factor-mediated immunity and interferon alpha (IFNA)-induced interferon-stimulated genes factor 3-mediated immunity, as shown in STAT1-deficient cells transfected with the corresponding alleles. Their phenotypic effects are however mediated by different molecular mechanisms, L706S affecting STAT1 phosphorylation and Q463H and E320Q affecting STAT1 DNA-binding activity. Heterozygous patients display specifically impaired IFNG-induced gamma-activating factor-mediated immunity, resulting in susceptibility to mycobacteria. Indeed, IFNA-induced interferon-stimulated genes factor 3-mediated immunity is not affected, and these patients are not particularly susceptible to viral disease, unlike patients homozygous for other, equally deleterious STAT1 mutations recessive for both phenotypes. The three STAT1 alleles are therefore dominant for IFNG-mediated antimycobacterial immunity but recessive for IFNA-mediated antiviral immunity at the cellular and clinical levels. These STAT1 alleles define two forms of dominant STAT1 deficiency, depending on whether the mutations impair STAT1 phosphorylation or DNA binding.

背景与目标： 转录因子信号转导子和转录激活因子1（STAT1）在抵抗分枝杆菌和病毒感染的免疫中起着关键作用。在这里，我们表征了来自其他健康的分枝杆菌病患者的三个人STAT1种系等位基因。像新的Q463H和E320Q等位基因一样，先前报道的L706S对干扰素γ（IFNG）诱导的γ激活因子介导的免疫和干扰素α（IFNA）诱导的干扰素刺激的基因因子3介导的免疫都具有内在的危害，如用相应等位基因转染的STAT1缺陷细胞所示。然而，它们的表型作用是由不同的分子机制介导的，L706S影响STAT1磷酸化，Q463H和E320Q影响STAT1 DNA结合活性。杂合子患者表现出特别受损的IFNG诱导的γ活化因子介导的免疫力，导致对分枝杆菌的易感性。确实，IFNA诱导的干扰素刺激基因因子3介导的免疫没有受到影响，并且这些患者对病毒性疾病特别不敏感，这与两种表型均隐性存在的其他同样有害的STAT1突变的患者不同。因此，在细胞和临床水平上，这三个STAT1等位基因在IFNG介导的抗分枝杆菌免疫中占主导地位，而在IFNA介导的抗病毒免疫中却处于隐性地位。这些STAT1等位基因定义了两种主要形式的STAT1缺陷，具体取决于突变是否损害STAT1磷酸化或DNA结合。

BACKGROUND & AIMS: :Tooth agenesis constitutes the most common anomaly of dental development in humans. In the majority of familial cases of hypodontia alone or in association with other anomalies, the mode of inheritance is autosomal dominant. In the present study, we have identified two distantly related consanguineous Pakistani kindreds with an autosomal recessive form of oligodontia with associated dental anomalies. Locus in this case has been mapped on chromosome 4p16.1-p16.3. The maximum two-point LOD score of 2.85 (theta=0.0) was obtained at markers D4S2925 and D4S2285. A maximum multipoint LOD score exceeding 4 was obtained at the same markers. Recombination events observed in affected individuals localized the disease locus between markers D4S412 and D4S2935, spanning a 9.24-cM region on chromosome 4p16.1-p16.3. Sequence analysis of candidate gene MSX1 revealed a novel recessive missense mutation resulting in substitution of alanine to threonine amino acid (p. A219T), located in the MSX1 homeodomain, which is important for DNA binding and protein-protein interaction. The mutation, p. A219T, is the first recessive mutation identified in MSX1.

背景与目标： 牙齿发育不全是人类牙齿发育的最常见异常。在大多数单独的或与其他异常有关的低血压的家族性病例中，遗传方式是常染色体显性的。在本研究中，我们确定了两个远缘近亲的巴基斯坦亲戚，他们是常染色体隐性形式的少齿畸形，并伴有相关的牙齿异常。在这种情况下，基因座已定位在染色体4p16.1-p16.3上。在标记D4S2925和D4S2285上获得的最大两点LOD得分为2.85（θ= 0.0）。在相同的标记处获得的最大多点LOD得分超过4。在受影响的个体中观察到的重组事件将疾病基因定位在标记D4S412和D4S2935之间，跨越染色体4p16.1-p16.3上的9.24-cM区。候选基因MSX1的序列分析揭示了一种新的隐性错义突变，该突变导致将丙氨酸替换为位于MSX1同源域的苏氨酸氨基酸（p。A219T），这对于DNA结合和蛋白质-蛋白质相互作用很重要。突变p。 A219T是MSX1中鉴定出的第一个隐性突变。

BACKGROUND & AIMS: :Differential killing of the patient's cancer cells versus normal cells is a necessity for chemotherapy. Advantage can be taken of close regulations of gene expression and of enzyme activity that are essential for normal cell functioning, and that are altered during tumor progression. Summarized here is our research on four such progression changes of cancer cells; some deregulate proliferation control and others decrease programmed death (apoptosis). These processes will be illustrated with examples of potential chemotherapies based on them. Methods for discovery of such changes include Differential Display and microarrays.

背景与目标： ：与正常细胞相比，不同程度地杀死患者的癌细胞是化疗的必要条件。可以利用基因表达和酶活性的紧密调节，这些调节对于正常的细胞功能是必不可少的，并且在肿瘤进展过程中会改变。这里总结了我们对癌细胞的四个这样的进展变化的研究。一些人放松了对增殖的控制，而另一些人减少了程序性死亡（细胞凋亡）。这些过程将以基于它们的潜在化学疗法为例进行说明。发现这种变化的方法包括差异显示和微阵列。

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： ：分布式基因组假设（DGH）指出，细菌物种中的每个菌株都从基于种群的超基因组中获得独特的基因分布，其比任何给定菌株的基因组都要大很多倍。自然感染人群通常是多克隆的，大多数慢性细菌病原体具有高度发展的水平基因转移机制的观察结果提示了DGH，并提供了解释哺乳动物宿主面对适应性防御机制时慢性感染如何持续存在的手段和机制。先前已确定DGH对专性病原体的有效性，我们希望评估其对机会细菌病原体的适用性。这是通过构建和分析包含约216,000个功能性克隆的高度冗余的集合基因组文库完成的，该文库由铜绿假单胞菌的12种低通道临床分离株，6种耳泻分离株和6个其他身体部位构建而成。对来自该文库的3,214个随机挑选的克隆（平均插入片段大小，约1.4 kb）进行序列分析，结果表明，对于铜绿假单胞菌原型菌株PAO1的所有基因组序列，其中348个（10.8％）克隆是唯一的。在这些独特序列内的开放阅读框的假想翻译证明了与许多细菌毒力因子和以前在铜绿假单胞菌中未鉴定出的其他蛋白质的蛋白质同源性。进行了基于PCR和逆转录PCR的分析，以分析这些序列的70个开放阅读框子集在11种临床菌株中的分布和表达模式。这些序列在临床分离株中分布不均，只有一半或34个新序列出现在一个或两个单独的菌株中。表达谱分析表明这些序列中的绝大多数都被表达，强烈暗示它们编码功能蛋白。

BACKGROUND & AIMS: :In a previous large scale screen for differentially expressed genes in pancreatic cancer, we identified a gene highly overexpressed in cancer encoding a novel protein with four K-homologous (KH) domains. KH-domains are found in a subset of RNA-binding proteins, including pre-mRNA-binding (hnRNP) K protein and the fragile X mental retardation gene product (FMR1). By fluorescence in situ hybridization (FISH) the identified gene named koc (KH domain containing protein overexpressed in cancer) was assigned to chromosome 7p11.5. Two pseudogenes were localised on chromosome 6 and 11. The cloned koc cDNA has a 250 bp 5'-UTR, a 1740 bp ORF and a 2168 bp 3'-UTR. The AU-rich 3'-untranslated region of koc contains eight AUUUA and four AUUUUUA reiterated motifs. The deduced koc protein with 580 amino-acids has a relative molecular mass (Mr) of approximately 65,000 (65 K). The koc transcript is highly overexpressed in pancreatic cancer cell lines and in pancreatic cancer tissue as compared to both, normal pancreas and chronic pancreatitis tissue. High levels of expression were as well found in tissue samples of other human tumours. As the KH domain has been shown to be involved in the regulation of RNA synthesis and metabolism, we speculate that koc may assume a role in the regulation of tumour cell proliferation by interfering with transcriptional and or posttranscriptional processes. However, the precise role of koc in human tumour cells is unknown and remains to be elucidated.

背景与目标： ：在先前针对胰腺癌中差异表达基因的大规模筛选中，我们鉴定了在癌症中高度过表达的基因，该基因编码具有四个K同源（KH）域的新型蛋白质。 KH结构域存在于RNA结合蛋白的子集中，包括前mRNA结合（hnRNP）K蛋白和脆弱的X智力低下基因产物（FMR1）。通过荧光原位杂交（FISH），将鉴定出的名为koc的基因（含有在癌症中过度表达的KH域蛋白）分配给7p11.5染色体。两个假基因位于6号和11号染色体上。克隆的koc cDNA具有250 bp的5'-UTR，1740 bp的ORF和2168 bp的3'-UTR。富含AU的3'非翻译区域的koc包含八个AUUUA和四个AUUUUUA重复的基序。推导的具有580个氨基酸的koc蛋白的相对分子质量（Mr）约为65,000（65 K）。与正常胰腺和慢性胰腺炎组织相比，koc转录本在胰腺癌细胞系和胰腺癌组织中高度过表达。在其他人类肿瘤的组织样本中也发现了高水平的表达。由于已经显示出KH结构域参与RNA合成和代谢的调节，我们推测koc可能通过干扰转录和/或转录后过程而在调节肿瘤细胞增殖中发挥作用。但是，koc在人肿瘤细胞中的确切作用尚不清楚，尚待阐明。

BACKGROUND & AIMS: :A cDNA encoding a novel human extracellularly-regulated kinase (ERK) phosphatase, designated B59, was isolated from a B5/589 human mammary epithelial cell cDNA library. The 1104 nucleotide open reading frame encodes 368 amino acids including the highly conserved catalytic site sequence of protein phosphotyrosine phosphatases (PTPs), VXVHCXXGXXR, at amino acid position 276-287. The predicted 70 amino acid stretch surrounding the HC motif shares significant sequence identity with other human dual specificity PTPs (dsPTPs), including the known ERK PTPs CL100, PAC1, B23, as well as the dsPTPs VH-1 and VHR. B59 protein synthesized in vitro in a rabbit reticulocyte lysate dephosphorylated rat ERK1 and ERK2 proteins whose phosphorylation had been stimulated by v-mos kinase added to the lysate. Ectopic expression of B59 in NIH3T3 fibroblasts inhibited the induction of an oncogene-responsive promoter by the dominant-activating raf mutant, raf-BXB. Moreover, cotransfection of NIH3T3 cells with B59 inhibited morphological transformation by H-ras and v-raf oncogenes. These results suggest that B59 suppresses the transforming activity of H-ras or v-raf oncogenes through ERK dephosphorylation and inactivation.

背景与目标： ：从B5 / 589人乳腺上皮细胞cDNA文库中分离出一种编码新型人胞外调节激酶（ERK）磷酸酶的cDNA，命名为B59。 1104个核苷酸的开放阅读框编码368个氨基酸，其中包括高度保守的蛋白质磷酸酪氨酸磷酸酶（PTP）催化位点序列VXVHCXXGXXR，位于氨基酸位置276-287。 HC基序周围的预测的70个氨基酸段与其他人的双特异性PTP（dsPTP），包括已知的ERK PTP CL100，PAC1，B23以及dsPTP VH-1和VHR，具有明显的序列同一性。在兔网织红细胞裂解物中体外合成的B59蛋白将磷酸化的大鼠ERK1和ERK2蛋白磷酸化，并通过添加到裂解物中的v-mos激酶刺激了磷酸化。 B59在NIH3T3成纤维细胞中的异位表达抑制了由显性激活raf突变体raf-BXB诱导的癌基因应答性启动子。此外，NIH3T3细胞与B59的共转染可抑制H-ras和v-raf癌基因的形态转化。这些结果表明，B59通过ERK去磷酸化和失活抑制H-ras或v-raf癌基因的转化活性。

BACKGROUND & AIMS: :Human immunodeficiency virus (HIV) binds to cells via an interaction between CD4 and the virus envelope glycoprotein, gp120. Previous studies have localized the high affinity binding site for gp120 to the first domain of CD4, and monoclonal antibodies (mAbs) reactive with this region compete with gp120 binding and thereby block virus infectivity and syncytium formation. Despite a detailed understanding of the binding of gp120 to CD4, little is known of subsequent events leading to membrane fusion and virus entry. We describe two new mAbs reactive with the third domain of CD4 that inhibit steps subsequent to virus binding critical for HIV infectivity and cell fusion. Binding of recombinant gp120 or virus to CD4 is not inhibited by these antibodies, whereas infection and syncytium formation by a number of HIV isolates are blocked. These findings demonstrate that in addition to virus binding, CD4 may have an active role in membrane fusion.

背景与目标： 人类免疫缺陷病毒（HIV）通过CD4和病毒包膜糖蛋白gp120之间的相互作用与细胞结合。先前的研究已将gp120的高亲和力结合位点定位在CD4的第一个域，并且与该区域反应的单克隆抗体（mAb）与gp120结合竞争，从而阻断了病毒的感染性和合胞体的形成。尽管对gp120与CD4的结合有了详细的了解，但对导致膜融合和病毒进入的后续事件知之甚少。我们描述了与CD4的第三个域具有反应性的两个新的单克隆抗体，可抑制继病毒结合后对HIV感染性和细胞融合至关重要的步骤。重组gp120或病毒与CD4的结合不受这些抗体的抑制，而许多HIV分离株的感染和合胞体形成却被阻止。这些发现表明，除病毒结合外，CD4可能在膜融合中发挥积极作用。

BACKGROUND & AIMS: :In a context of automation of cryo-electron microscopy, we developed a novel method for improving visibility of diffraction rings in the power spectra of cryo-electron micrographs of vitreous ice (without carbon film or high concentration of diffracting material). We used these enhanced spectra to semi-automatically detect and remove micrographs and/or local areas introducing errors in the global 3D map (drifted and charged areas) or those unable to increase global signal-to-noise ratio (non-diffracting areas). Our strategy also allows a detection of micrographs/areas with a strong astigmatism. These images should be removed when using algorithms that do not correct astigmatism. Our sorting method is simple and fast since it uses the normalized cross-correlation between enhanced spectra and their copies rotated by 90 degrees. It owes its success mainly to the novel pre-processing of power spectra. The improved visibility also allows an easier visual check of accuracy of sorting. We show that our algorithm can even improve the visibility of diffraction rings of cryo-electron micrographs of pure water. Moreover, we show that this visibility depends strongly on ice thickness. This algorithm is implemented in the Xmipp (open-source image processing package) and is freely available for implementation in any other software package.

背景与目标： ：在低温电子显微镜自动化的背景下，我们开发了一种新颖的方法，用于改善玻璃冰（无碳膜或高浓度衍射材料）的低温电子显微镜照片的功率谱中衍射环的可见性。我们使用这些增强的光谱来半自动检测和移除显微照片和/或局部区域，从而在全局3D地图（漂移和带电区域）或无法增加全局信噪比（非衍射区域）的区域引入误差。我们的策略还允许检测具有强烈散光的显微照片/区域。当使用无法校正像散的算法时，应删除这些图像。我们的排序方法简单而快速，因为它使用了增强频谱与其旋转90度的副本之间的归一化互相关。它的成功主要归功于新颖的功率谱预处理。改进的可见性还可以使目视检查的准确性更加容易。我们证明了我们的算法甚至可以提高纯水冷冻电子显微照片的衍射环的可见性。此外，我们证明了这种能见度在很大程度上取决于冰的厚度。该算法在Xmipp（开源图像处理软件包）中实现，可免费用于任何其他软件包中。

BACKGROUND & AIMS: :Large-scale copy number variation that is cytogenetically visible in normal individuals has been described as euchromatic variation but needs to be distinguished from pathogenic euchromatic deletion or duplication. Here, we report eight patients (three families and two individuals) with interstitial deletions of 9q13-q21.12. Fluorescence in situ hybridisation with a large panel of BACs showed that all the deleted clones were from extensive tracts of segmentally duplicated euchromatin, copies of which map to both the long and short arms of chromosome 9. The variety of reasons for which these patients were ascertained, and the phenotypically normal parents, indicates that this is a novel euchromatic variant with no phenotypic effect. Further, four patients with classical euchromatic variants of 9q12/qh or 9p12 were also shown to have duplications or triplications of this segmentally duplicated material common to both 9p and 9q. The cytogenetic boundaries between the segmentally duplicated regions and flanking unique sequences were mapped to 9p13.1 in the short arm (BAC RP11-402N8 at 38.7 Mb) and to 9q21.12 in the long arm (BAC RP11-88I18 at 70.3 Mb). The BACs identified in this study should in future make it possible to differentiate between clinically significant deletions or duplications and euchromatic variants with no established phenotypic consequences.

背景与目标： ：在正常个体中在细胞遗传学上可见的大规模拷贝数变异已被描述为常染色体变异，但需要与致病性常染色体缺失或复制区分开。在这里，我们报告8例患者（3个家庭和2个个体）的间质缺失9q13-q21.12。与大量BAC的荧光原位杂交表明，所有缺失的克隆均来自节段重复的常染色质的广泛片段，其拷贝定位于9号染色体的长臂和短臂。确定这些患者的各种原因和表型正常的父母，表明这是一个新的常染色体变异，没有表型作用。此外，还显示了四名具有9q12 / qh或9p12的经典常染色体变体的患者具有这种重复出现或重复的9p12和9q共同的节段重复材料。片段重复区域和侧翼独特序列之间的细胞遗传学边界在短臂中定位为9p13.1（BAC RP11-402N8，38.7 Mb），在长臂中定位9q21.12，长臂（BAC RP11-88I18，70.3 Mb）。这项研究中确定的BAC将来应该可以区分具有临床意义的缺失或重复和常染色体变异，而没有确定的表型后果。

BACKGROUND & AIMS: :A major problem in the search for new cancer drug targets is that the drugs are often toxic to normal tissues and require high doses to kill tumor cells. Therefore cellular targets which appear to involve low dose responses to cancer therapy are especially interesting since they could selectively target normal tissues which are not targeted by the treatment and thus may be responsible for unpleasant side effects or may be amenable to exploitation in order to improve the therapeutic ratio. One such target, which is the subject of this review, is radiation-induced bystander effects [RIBE], which result in the observation of radiation like responses in cells which have not been irradiated. RIBE is a novel phenomenon which indicates that at low doses, cell signaling is more important than direct DNA damage. Historically, DNA has always been considered to be the target for radiation therapy. The growing realization that signaling is important opens up several important therapeutic strategies which will be discussed in this review. RIBE appears to be the result of a generalized stress response in tissues or cells which is expressed at the level of the tissue, organ or organism rather than at the level of the individual cell. The signals may be produced by all exposed cells, but the response may require a quorum of cells in order to be expressed. The major response involving low LET (x- or gamma-ray) radiation exposure discussed in the existing literature is a death response. This has many characteristics of apoptosis but may be detected in cell lines without p53 expression, although the death response is suppressed in many tumor cell lines. While a death response in unirradiated normal cells around a tumor might appear to be adverse, it can in fact be protective and remove damaged cells from the population. If harnessed correctly, it could lead to the development of new drugs aimed not at tissue destruction but at enabling homeostatic mechanisms to control tumor expansion. In this scenario, the level of harmful or beneficial response will be related to the background damage, carried by the cell population, and the genetic programme determining response to damage. This focus may be important when attempting to predict the consequences of mixed therapies involving radiation and other cytotoxic agents. In this review, our current knowledge of the mechanisms underlying the induction of bystander effects by ionizing radiation is reviewed, and the question of how bystander effects may be harnessed to produce a new generation of anti-cancer drugs aimed at stabilization of tissue homeostasis rather than tissue destruction is considered.

背景与目标： ：寻找新的癌症药物靶标的主要问题是该药物通常对正常组织有毒性，需要高剂量才能杀死肿瘤细胞。因此，细胞靶标似乎涉及对癌症治疗的低剂量反应，因此特别令人感兴趣，因为它们可以选择性地靶向未被治疗靶标的正常组织，因此可能引起令人不快的副作用，或者可能适于利用以改善治疗效果。治疗比率。辐射诱导的旁观者效应[RIBE]是本综述的主题之一，该效应导致在未辐射的细胞中观察到辐射样反应。 RIBE是一种新现象，表明在低剂量时，细胞信号传导比直接DNA损伤更为重要。从历史上看，DNA一直被认为是放射治疗的目标。人们日益认识到信号转导很重要，这开启了几种重要的治疗策略，本文将对此进行讨论。 RIBE似乎是组织或细胞中普遍的应激反应的结果，这种应激反应是在组织，器官或生物体的水平而不是单个细胞的水平表达的。信号可能由所有暴露的细胞产生，但响应可能需要一定数量的细胞才能表达。现有文献中讨论的涉及低LET（X射线或γ射线）辐射暴露的主要反应是死亡反应。这具有许多细胞凋亡特征，但尽管在许多肿瘤细胞系中死亡反应受到抑制，但在没有p53表达的细胞系中可能检测到。虽然在肿瘤周围未照射的正常细胞中的死亡反应似乎是不利的，但实际上可以起到保护作用，并从群体中清除受损的细胞。如果利用得当，它可能会导致开发新药物，其目的不是破坏组织，而是使稳态机制能够控制肿瘤的扩展。在这种情况下，有害或有益反应的水平将与细胞群体所携带的背景损伤以及决定对损伤的反应的遗传程序有关。当试图预测涉及放射线和其他细胞毒剂的混合疗法的后果时，这一重点可能很重要。在这篇综述中，我们对电离辐射诱发旁观者效应的潜在机制的现有知识进行了综述，并探讨了如何利用旁观者效应来生产旨在稳定组织稳态而不是稳定组织的新一代抗癌药物的问题。考虑组织破坏。

BACKGROUND & AIMS: :The pharmacological profile of MEN15596 or (6-methyl-benzo[b]thiophene-2-carboxylic acid [1-(2-phenyl-1R-{[1-(tetrahydropyran-4-ylmethyl)-piperidin-4-ylmethyl]-carbamoyl}-ethylcarbamoyl)-cyclopentyl]-amide), a novel potent and selective tachykinin NK2 receptor antagonist endowed with oral activity, is described. At the human recombinant tachykinin NK2 receptor, MEN15596 showed subnanomolar affinity (pKi 10.1) and potently antagonized (pKB 9.1) the neurokinin A-induced intracellular calcium release. MEN15596 selectivity for the tachykinin NK2 receptor was assessed by binding studies at the recombinant tachykinin NK1 (pKi 6.1) and NK3 (pKi 6.4) receptors, and at a number of 34 molecular targets including receptors, transporters and ion channels. In isolated smooth muscle preparations MEN15596 showed a marked species selectivity at the tachykinin NK2 receptor with the highest antagonist potency in guinea-pig colon, human and pig bladder (pKB 9.3, 9.2 and 8.8, respectively) whereas it was three orders of magnitude less potent in the rat and mouse urinary bladder (pKB 6.3 and 5.8, respectively). In agreement with binding experiments, MEN15596 showed low potency in blocking selective NK1 or NK3 receptor agonist-induced contractions of guinea-pig ileum preparations (pA2

BACKGROUND & AIMS: :The human iodotyrosine dehalogenase 1 (DEHAL1) gene is composed of six exons. Two isoforms (DEHAL1 and DEHAL1B) have been published in GenBank, both of which have a nitroreductase domain and arise from differential splicing in exon 5. We recently showed that the DEHAL1 isoform is a transmembrane protein that efficiently catalyzes the NADPH-dependent deiodination of mono (L-MIT) and diiodotyrosine (L-DIT) in human embryonic kidney-293 (HEK293) cells. In the present study, we establish the existence of a new transcript, DEHAL1C, in the human thyroid with a terminal exon that lacks in the DEHAL1 transcript. This exon is the complete exon 5, which is spliced in the DEHAL1B mRNA variant. These two variants encode proteins with differing C-terminal domains. Using quantitative reverse transcription polymerase chain reaction, we found that the expression of the mRNA of DEHAL1C and DEHAL1B was lower than that of DEHAL1 mRNA in the thyroid. We also observed that human DEHAL1B and DEHAL1C proteins are rapidly degraded in stably transfected HEK293 cells, unlike the DEHAL1 protein, and that exposure to the proteasome inhibitor MG132 resulted in accumulation of these proteins that was markedly time- and concentration-dependent. These findings show that the cytoplasmic tail could play a role in the stability of the protein.

背景与目标： ：人碘酪氨酸脱卤素酶1（DEHAL1）基因由六个外显子组成。 GenBank中已公开了两种同工型（DEHAL1和DEHAL1B），它们均具有硝基还原酶结构域，并来自外显子5的差异剪接。 （L-MIT）和二碘代酪氨酸（L-DIT）在人类胚胎肾293（HEK293）细胞中。在本研究中，我们在人甲状腺中建立了一个新的转录物DEHAL1C的存在，其末端外显子缺少DEHAL1转录物。该外显子是完整的外显子5，其剪接在DEHAL1B mRNA变异体中。这两个变体编码具有不同C末端结构域的蛋白质。通过定量逆转录聚合酶链反应，我们发现甲状腺中DEHAL1C和DEHAL1B的mRNA表达低于DEHAL1 mRNA的表达。我们还观察到，与DEHAL1蛋白不同，人DEHAL1B和DEHAL1C蛋白在稳定转染的HEK293细胞中迅速降解，并且暴露于蛋白酶体抑制剂MG132导致这些蛋白的积累具有明显的时间和浓度依赖性。这些发现表明，胞质尾部可以在蛋白质的稳定性中起作用。

BACKGROUND & AIMS: :Inborn errors of vitamin B(12) (cobalamin) metabolism are characterized by decreased production of active cobalamin cofactors and subsequent deficiencies in the activities of methionine synthase and methylmalonyl-CoA mutase. With the recent discovery of the cblJ defect in two patients with phenotypes mimicking the cblF defect, there are nine genes known to be involved in cobalamin metabolism. The new defect is caused by mutations in the ABCD4 gene, encoding an ABC transporter. At the moment, there is no clear distinction between the cblJ and cblF defects either clinically or biochemically, and both defects result in blocks in the transport of cobalamin from the lysosome to the cytoplasm. A patient was diagnosed with hyperhomocysteinemia and methylmalonic aciduria at the age of 8 years. Incorporations of both [(14)C]propionate and [(14)C]methyltetrahydrofolate in cultured fibroblasts were within reference ranges and thus too high to allow for complementation analysis. We observed decreased synthesis of both adenosylcobalamin and methylcobalamin and accumulation of unmetabolized cyanocobalamin. Exome sequencing was performed to identify causative mutation(s) and Sanger re-sequencing was performed to validate segregation of mutation in the family. By this approach, a homozygous mutation, c.423C>G, in the ABCD4 gene was identified. Here, we report the successful application of exome sequencing for diagnosis of a rare inborn error of vitamin B(12) metabolism in a patient whose unusual presentation precluded diagnosis using standard biochemical and genetic approaches. The patient represents only the third known patient with the cblJ disorder.

背景与目标： ：维生素B（12）（钴胺素）代谢的先天性错误的特征在于活性钴胺素辅因子的产生减少以及蛋氨酸合酶和甲基丙二酰辅酶A突变酶的活性随后不足。随着最近在两名模仿cblF缺陷的表型患者中发现cblJ缺陷，已知有9个基因与钴胺素代谢有关。新的缺陷是由编码ABC转运蛋白的ABCD4基因突变引起的。目前，在临床或生化方面，cblJ和cblF缺陷之间尚无明确区分，并且两种缺陷均导致钴胺素从溶酶体到细胞质的转运受阻。一名患者在8岁时被诊断出患有高同型半胱氨酸血症和甲基丙二酸尿症。 [（14）C]丙酸酯和[（14）C]甲基四氢叶酸在培养的成纤维细胞中的掺入均在参考范围内，因此含量过高，无法进行互补分析。我们观察到腺苷钴胺素和甲基钴胺素的合成减少以及未代谢的氰钴胺素的积累。进行了外显子组测序以鉴定致病突变，并进行了桑格重测序以验证家族中突变的分离。通过这种方法，鉴定出ABCD4基因中的纯合突变，即c.423C> G。在这里，我们报告外显子组测序在维生素B（12）代谢的罕见先天性错误的诊断中成功应用，该患者的异常表现排除了使用标准生化和遗传方法进行诊断的可能性。该患者仅代表第三位已知的cblJ疾病患者。

BACKGROUND & AIMS: :Silent information regulator 2 (Sir2) enzymes or sirtuins are a family of NAD(+)-dependent protein N(ε)-acetyl-lysine (AcK) deacetylases. Sirtuins are also evolutionarily conserved proteins that are present in all kingdoms of life ranging from bacteria to humans. Interestingly, it was recently found that the sirtuins found in various human parasites (especially the Plasmodium, Trypanosoma, and Leishmania species) were pro-survival for the parasites under various conditions. Therefore, these parasitic sirtuins have emerged as novel anti-parasitic therapeutic targets. This article reviews the currently available structural, biochemical, pharmacological, and medicinal chemistry studies on these enzymes, and discusses the perspectives of selectively targeting the parasitic sirtuins as a novel therapeutic strategy for the human parasitic diseases.

背景与目标： ：沉默信息调节剂2（Sir2）酶或sirtuins是NAD（）依赖性蛋白N（ε）-乙酰赖氨酸（AcK）脱乙酰基酶的一个家族。 Sirtuins还是进化上保守的蛋白质，存在于从细菌到人类的所有生命王国中。有趣的是，最近发现在各种人类寄生虫（尤其是疟原虫，锥虫和利什曼原虫）中发现的沉默调节蛋白在各种条件下均可存活。因此，这些寄生沉默调节蛋白已经成为新型的抗寄生虫治疗靶标。本文回顾了有关这些酶的当前可用的结构，生化，药理和药物化学研究，并讨论了选择性靶向寄生虫沉默调节蛋白作为人类寄生虫疾病的新型治疗策略的观点。