BACKGROUND & AIMS: The human autosomal dominant neuromuscular disorder facioscapulohumeral muscular dystrophy (FSHD) is associated with deletions within a complex tandem DNA repeat (D4Z4) on Chromosome (Chr) 4q35. The molecular mechanism underlying this association of FSHD with DNA rearrangements is unknown, and, thus far, no gene has been identified within the repeat. We isolated a gene mapping 100 kb proximal to D4Z4 (FSHD Region Gene 1FRG1), but were unable to detect any alterations in total or allele-specific mRNA levels of FRG1 in FSHD patients. Human Chr 4q35 exhibits synteny homology with the region of mouse Chr 8 containing the gene for the myodystrophy mutation (myd), a possible mouse homolog of FSHD. We report the cloning of the mouse gene (Frg1) and show that it maps to mouse Chr 8. Using a cross segregating the myd mutation and the European Collaborative Interspecific Backcross, we showed that Frg1 maps proximal to the myd locus and to the Clc3 and Ant1 genes.

背景与目标： 人类常染色体显性遗传性神经肌肉疾病面肩肱肱型肌营养不良症（FSHD）与染色体（Chr）4q35上复杂的串联DNA重复序列（D4Z4）内的缺失相关。 FSHD与DNA重排相关的分子机制尚不清楚，到目前为止，在重复序列中尚未鉴定出任何基因。我们分离了一个基因，定位于D4Z4（FSHD区域基因1FRG1）近100 kb的基因，但无法检测到FSHD患者FRG1的总或等位基因特异性mRNA水平有任何改变。人Chr 4q35与小鼠Chr 8区域具有同系同源性，该区域包含肌营养不良症突变（myd）的基因，该基因可能是FSHD的小鼠同源物。我们报告了小鼠基因（Frg1）的克隆，并显示其映射到小鼠Chr8。使用myd突变和欧洲合作种间回交的交叉分离，我们显示Frg1映射到myd基因座以及Clc3和Ant1基因。

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: :Plant leaves are a crucial organ associated closely with chloroplast development, photosynthesis rate and crop productivity. In this study, a white fine stripe leaf 1 (wfsl1) mutant was isolated and characterized from the japonica rice Zhonghua11 (ZH11) after ethyl methanesulfonate mutagenesis. The wfsl1 displayed white fine stripe leaves since tillering stage and abnormal chloroplast structure. Map-based cloning and Bioinformatic analysis indicated that WFSL1 on chromosome 1 contains an "A" to "T" substitution in protein coding region, and encodes a putative metal-dependent phosphohydrolase with HD domain at the N-terminus. WFSL1 was targeted to the chloroplasts and had higher expression in mature leaves and sheaths. RNA-seq analysis revealed that chloroplast development and photosynthesis genes were significantly affected in wfsl1 plants. Levels of WFSL1 and chloroplast encoded proteins were decreased in wfsl1 mutants via western blot analysis. Compared with WT, wfsl1 exhibits lower Chl content and defective in biogenesis of chloroplast ribosomes, which resulted in reduced grain yield. Taken together, our results show that WFSL1 is critical for chloroplast development, ribosome biogenesis, and light energy utilization, finally affects grain yield.

背景与目标： 植物叶片是与叶绿体发育，光合作用速率和农作物生产力密切相关的重要器官。在本研究中，从甲磺酸乙酯诱变后的粳稻中华11（ZH11）中分离并鉴定了白色细条叶1（wfsl1）突变体。分f期和叶绿体结构异常后，wfsl1呈白色细条状叶片。基于图谱的克隆和生物信息学分析表明，染色体1上的WFSL1在蛋白质编码区中包含“ A”至“ T”取代，并在N端编码具有HD结构域的推定的金属依赖性磷酸水解酶。 WFSL1靶向叶绿体，并在成熟的叶和鞘中具有较高的表达。 RNA-seq分析显示，wfsl1植物的叶绿体发育和光合作用基因受到显着影响。通过Western blot分析，wfsl1突变体中WFSL1和叶绿体编码蛋白的水平降低。与野生型相比，wfsl1的Chl含量较低，叶绿体核糖体的生物发生缺陷，从而导致谷物产量下降。综上所述，我们的结果表明WFSL1对叶绿体发育，核糖体生物发生和光能利用至关重要，最终影响谷物产量。

BACKGROUND & AIMS: The mechanism of recombination of tandem repeats in the chromosome of Escherichia coli was investigated by genetic means. Tandem repeats 624 bp long were introduced into the lacZ gene of E. coli and the efficiency of deletion of one repeat was compared in different recombination mutants. No effects of the recA, recBC, recF, ruvA or ruvA recG mutations were detected. Hence, tandem repeat deletion appears to not proceed via the RecBCD or RecF homologous recombination pathways. A new mutant in which RecA-independent recombination is increased 15-fold was isolated. The mutation lies in the dnaE gene coding for the alpha subunit of polymerase IIIit is a Gly to Asp change at codon 133. Another dnaE mutation, dnaE486, was tested and also shown to stimulate RecA-independent recombination. It is proposed that tandem-repeat recombination occurs by a replication slippage mechanism. RecA-independent recombination is also enhanced in a rep mutant, in which chromosomal replication is slowed down by the absence of the Rep helicase, suggesting that replication pausing may facilitate slippage.

背景与目标： 通过遗传手段研究了大肠杆菌染色体上串联重复序列的重组机制。将624bp长的串联重复序列引入到大肠杆菌的lacZ基因中，并比较了不同重组突变体中一个重复序列的缺失效率。未检测到recA，recBC，recF，ruvA或ruvA recG突变的影响。因此，串联重复删除似乎不会通过RecBCD或RecF同源重组途径进行。分离出一个新的突变体，其中不依赖RecA的重组增加了15倍。该突变位于编码聚合酶IIIit的α亚基的dnaE基因，它是第133位密码子由Gly变为Asp的另一种dnaE突变，即dnaE486，经过测试，也显示出可刺激与RecA无关的重组。建议通过复制滑动机制发生串联-重复重组。不依赖RecA的重组在rep突变体中也得到增强，其中由于Rep解旋酶的缺乏而减慢了染色体的复制，这表明复制暂停可能有助于滑动。

BACKGROUND & AIMS: :The identification of germ-line mutations in 2 genes (BRCA1 and BRCA2) responsible for the majority of hereditary ovarian cancers has led an increasing number of women carriers of these mutations to undergo prophylactic oophorectomy (PO) to reduce their risk of subsequent ovarian carcinoma. A large number of unexpected, clinically occult neoplasms are thus being discovered. Up to December 2004, the Medical Genetics Service of the National Cancer Institute in Milan, Italy, has tested 756 probands from breast and/or ovarian cancer families for BRCA1 and BRCA2 germ-line mutations. Molecular screening of family members led to the identification of 344 female carriers of BRCA1 (239) or BRCA2 (105) germ-line mutations. Of the 186 potentially eligible women (37 of whom had tested positive for BRCA1 and 13 for BRCA2 mutation), 50 (26.8%) chose to undergo PO. Six clinically occult primary gynecologic malignancies (2 stage IIIC serous carcinomas of the ovary, 3 in situ serous carcinomas of the fallopian tube, and 1 stage IIB invasive serous carcinoma of the fallopian tube) and 1 occult ovarian metastasis from breast carcinoma were identified in the PO specimens of 7 women (all BRCA1 mutated). Four of the patients with occult primary gynecologic cancers are alive without disease 129, 87, 38, and 7 months after PO, respectively. One of the 2 patients with primary ovarian cancer and the single patient with tubal invasive carcinoma are alive with recurrent disease 83 and 20 months after PO, respectively. In addition, one of the patients whose PO specimen did not show any malignancy presented with stage IIIC tubal carcinoma 77 months after PO. The relatively high number of tubal neoplasms found at PO in this group of patients underlines the linkage between mutation and the risk of developing tubal cancer, and stresses the need to include removal of the entire tubes at the time of PO and of thoroughly evaluating the specimens at the microscopic level. The upstaging of all 3 invasive carcinomas after staging surgery, and the late recurrence and persistence of 2 of them despite treatment indicate that small size of the tumors should not preclude therapy.

背景与目标： ：对负责大多数遗传性卵巢癌的2个基因（BRCA1和BRCA2）的种系突变的鉴定已导致越来越多的女性携带这些突变的女性进行预防性卵巢切除术（PO），以降低其患上卵巢癌的风险。因此，发现了大量意想不到的临床隐匿性肿瘤。截至2004年12月，意大利米兰国家癌症研究所的医学遗传学服务已对756个来自乳腺癌和/或卵巢癌家族的先证者进行了BRCA1和BRCA2种系突变测试。家庭成员的分子筛查导致鉴定出344例BRCA1（239）或BRCA2（105）种系突变的女性携带者。在186名可能符合条件的妇女中（其中37人的BRCA1测试呈阳性，13人的BRCA2突变检测为阳性），其中50人（26.8％）选择接受PO。在临床中确定了6例临床隐匿的原发性妇科恶性肿瘤（2例卵巢IIIC浆液性癌，3例输卵管原位浆液性癌和1例IIB输卵管浸润性浆液性癌）和1例隐匿性卵巢癌卵巢转移。 7名妇女的PO标本（所有BRCA1突变）。分别在PO后129、87、38和7个月，有四名患有隐匿性原发性妇科癌症的患者还活着而没有疾病。 2例原发性卵巢癌患者中的1例和输卵管浸润性癌的单例患者在PO后分别存活83个月和20个月。此外，其中一名PO标本未显示任何恶性肿瘤的患者在PO后77个月出现IIIC期输卵管癌。该组患者在PO中发现的相对较多的输卵管肿瘤强调了突变与发生输卵管癌的风险之间的联系，并强调需要在PO时切除整个管并彻底评估标本在微观层面上。分期手术后所有3种浸润性癌的分期升级，并且尽管有治疗，但其中2种仍较晚复发和持续存在，这表明较小的肿瘤不应排除治疗的可能性。

BACKGROUND & AIMS: :Despite a growing number of splicing mutations found in hereditary diseases, utilization of aberrant splice sites and their effects on gene expression remain challenging to predict. We compiled sequences of 346 aberrant 5'splice sites (5'ss) that were activated by mutations in 166 human disease genes. Mutations within the 5'ss consensus accounted for 254 cryptic 5'ss and mutations elsewhere activated 92 de novo 5'ss. Point mutations leading to cryptic 5'ss activation were most common in the first intron nucleotide, followed by the fifth nucleotide. Substitutions at position +5 were exclusively G>A transitions, which was largely attributable to high mutability rates of C/G>T/A. However, the frequency of point mutations at position +5 was significantly higher than that observed in the Human Gene Mutation Database, suggesting that alterations of this position are particularly prone to aberrant splicing, possibly due to a requirement for sequential interactions with U1 and U6 snRNAs. Cryptic 5'ss were best predicted by computational algorithms that accommodate nucleotide dependencies and not by weight-matrix models. Discrimination of intronic 5'ss from their authentic counterparts was less effective than for exonic sites, as the former were intrinsically stronger than the latter. Computational prediction of exonic de novo 5'ss was poor, suggesting that their activation critically depends on exonic splicing enhancers or silencers. The authentic counterparts of aberrant 5'ss were significantly weaker than the average human 5'ss. The development of an online database of aberrant 5'ss will be useful for studying basic mechanisms of splice-site selection, identifying splicing mutations and optimizing splice-site prediction algorithms.

背景与目标： ：尽管在遗传性疾病中发现了越来越多的剪接突变，但异常剪接位点的利用及其对基因表达的影响仍然难以预测。我们编辑了346个异常5'剪接位点（5个）的序列，这些序列被166个人类疾病基因的突变激活。 5位共识中的突变占254个隐性5位，而其他地方的突变则激活了92位从头5位。导致隐伏5激活的点突变最常见于第一个内含子核苷酸，其次是第五个核苷酸。位置5的替换仅是G> A转换，这主要归因于C / G> T / A的高变异率。但是，第5位的点突变频率显着高于人类基因突变数据库中观察到的频率，这表明该位置的改变特别容易出现异常剪接，这可能是由于需要与U1和U6 snRNA进行顺序相互作用所致。隐秘5最好通过适应核苷酸依赖性的计算算法进行预测，而不是通过权重矩阵模型进行预测。与前者相比，将内含子5与真正的异物区别开来的效果不佳，因为前者在本质上要强于后者。新外显子5's的计算预测很差，表明它们的激活关键取决于外显子剪接增强剂或沉默子。异常5的真实对应物显着弱于普通人类5的。异常5的在线数据库的开发将对研究剪接位点选择的基本机制，鉴定剪接突变和优化剪接位点预测算法非常有用。

BACKGROUND & AIMS: :This study investigated the dynamics of Aspergillus fumigatus azole-resistant phenotypes in two compost heaps with contrasting azole exposures: azole free and azole exposed. After heat shock, to which sexual but not asexual spores are highly resistant, the azole-free compost yielded 98% (49/50) wild-type and 2% (1/50) azole-resistant isolates, whereas the azole-containing compost yielded 9% (4/45) wild-type and 91% (41/45) resistant isolates. From the latter compost, 80% (36/45) of the isolates contained the TR46/Y121F/T289A genotype, 2% (1/45) harbored the TR46/Y121F/M172I/T289A/G448S genotype, and 9% (4/45) had a novel pan-triazole-resistant mutation (TR463/Y121F/M172I/T289A/G448S) with a triple 46-bp promoter repeat. Subsequent screening of a representative set of clinical A. fumigatus isolates showed that the novel TR463 mutant was already present in samples from three Dutch medical centers collected since 2012. Furthermore, a second new resistance mutation was found in this set that harbored four TR46 repeats. Importantly, in the laboratory, we recovered the TR463 mutation from a sexual cross between two TR46 isolates from the same azole-containing compost, possibly through unequal crossing over between the double tandem repeats (TRs) during meiosis. This possible role of sexual reproduction in the emergence of the mutation was further implicated by the high level of genetic diversity of STR genotypes in the azole-containing compost. Our study confirms that azole resistance mutations continue to emerge in the environment and indicates compost containing azole residues as a possible hot spot. Better insight into the biology of environmental resistance selection is needed to retain the azole class for use in food production and treatment of Aspergillus diseases.IMPORTANCE Composting of organic matter containing azole residues might be important for resistance development and subsequent spread of resistance mutations in Aspergillus fumigatus In this article, we show the dominance of azole-resistant A. fumigatus in azole-exposed compost and the discovery of a new resistance mutation with clinical relevance. Furthermore, our study indicates that current fungicide application is not sustainable as new resistance mutations continue to emerge, thereby threatening the use of triazoles in medicine. We provide evidence that the sexual part of the fungal life cycle may play a role in the emergence of resistance mutations because under laboratory conditions, we reconstructed the resistance mutation through sexual crossing of two azole-resistant A. fumigatus isolates derived from the same compost heap. Understanding the mechanisms of resistance selection in the environment is needed to design strategies against the accumulation of resistance mutations in order to retain the azole class for crop protection and treatment of Aspergillus diseases.

背景与目标： ：这项研究调查了两个堆肥堆中烟熏曲霉的耐唑性表型的动态变化，这两种情况下与吡咯暴露的情况相反：无吡咯和暴露于吡咯。在热休克后，对性但非无性孢子具有很高的抵抗力，无唑堆肥产生98％（49/50）野生型和2％（1/50）对唑的抗药性，而含吡咯的堆肥产生9％（4/45）野生型和91％（41/45）抗性分离株。在后一种堆肥中，80％（36/45）的分离株含有TR46 / Y121F / T289A基因型，2％（1/45）具有TR46 / Y121F / M172I / T289A / G448S基因型，9％（4 / 45）具有新的泛三唑抗性突变（TR463 / Y121F / M172I / T289A / G448S），具有三重46 bp启动子重复序列。随后对代表性的临床烟曲霉菌株进行了筛选，结果显示自2012年以来从三个荷兰医学中心收集的样品中已经存在新的TR463突变体。此外，在该菌株中发现了第二个新的耐药突变，其中包含四个TR46重复序列。重要的是，在实验室中，我们可能是通过减数分裂过程中双串联重复序列（TR）之间的不相等交换，从同一个含唑的堆肥中的两个TR46分离株之间的有性杂交中恢复了TR463突变。在含唑的堆肥中，STR基因型的高遗传多样性进一步暗示了性繁殖在突变出现中的这种可能作用。我们的研究证实，环境中仍会继续出现唑类抗性突变，并表明含有唑类残留物的堆肥可能是热点。需要更好地了解环境抗性选择的生物学以保持唑类用于食品生产和治疗曲霉病。重要事项含吡咯残留物的有机物的堆肥对于烟曲霉的抗性发展和随后的抗性突变的传播可能很重要。在本文中，我们显示了对吡唑有抵抗力的烟曲霉在暴露有吡咯的堆肥中的优势地位，并发现了与临床相关的新的抗药性突变。此外，我们的研究表明，随着新的耐药性突变的不断出现，当前杀菌剂的应用是不可持续的，从而威胁到三唑类药物的使用。我们提供的证据表明，真菌生命周期的性部分可能在耐药性突变的出现中起作用，因为在实验室条件下，我们通过对两个具有唑类抗性的A进行有性杂交来重建耐药性突变。来自同一堆肥堆的烟熏分离株。需要了解环境中抗性选择的机制，以设计针对抗性突变积累的策略，以保留用于农作物保护和曲霉病治疗的唑类。

BACKGROUND & AIMS: :Chronic exposure to high glucose leads to diabetic nephropathy characterized by increased mesangial matrix protein (e.g., collagen) accumulation. Altered cell signaling and gene expression accompanied by oxidative stress have been documented. The contribution of the tyrosine kinase, c-Src (Src), which is sensitive to oxidative stress, was examined. Cultured rat mesangial cells were exposed to high glucose (25 mmol/L) in the presence and absence of Src inhibitors (PP2, SU6656), Src small interfering RNA (siRNA), and the tumor necrosis factor-α-converting enzyme (TACE) inhibitor, TAPI-2. Src was investigated in vivo by administration of PP2 to streptozotocin (STZ)-induced diabetic DBA2/J mice. High glucose stimulated Src, TACE, epidermal growth factor receptor (EGFR), mitogen-activated protein kinases (MAPKs), extracellular signal-regulated kinase (ERK1/2, p38), and collagen IV accumulation in mesangial cells. PP2 and SU6656 blocked high glucose-stimulated phosphorylation of Src Tyr-416, EGFR, and MAPKs. These inhibitors and Src knockdown by siRNA, as well as TAPI-2, also abrogated high glucose-induced phosphorylation of these targets and collagen IV accumulation. In STZ-diabetic mice, albuminuria, increased Src pTyr-416, TACE activation, ERK and EGFR phosphorylation, glomerular collagen accumulation, and podocyte loss were inhibited by PP2. These data indicate a role for Src in a high glucose-Src-TACE-heparin-binding epidermal growth factor-EGFR-MAPK-signaling pathway to collagen accumulation. Thus, Src may provide a novel therapeutic target for diabetic nephropathy.

背景与目标： ：长期暴露于高葡萄糖会导致糖尿病肾病，其特征是肾小球系膜基质蛋白（例如胶原蛋白）蓄积增加。已经记录了改变的细胞信号传导和基因表达并伴有氧化应激。检查了对氧化应激敏感的酪氨酸激酶c-Src（Src）的贡献。在存在和不存在Src抑制剂（PP2，SU6656），Src小干扰RNA（siRNA）和肿瘤坏死因子-α转化酶（TACE）的情况下，将培养的大鼠肾小球系膜细胞暴露于高葡萄糖（25 mmol / L）抑制剂，TAPI-2。通过对链脲佐菌素（STZ）诱导的糖尿病DBA2 / J小鼠施用PP2在体内研究了Src。高葡萄糖刺激的Src，TACE，表皮生长因子受体（EGFR），有丝分裂原激活的蛋白激酶（MAPK），细胞外信号调节激酶（ERK1 / 2，p38）和胶原IV在肾小球膜细胞中的蓄积。 PP2和SU6656阻止了Src Tyr-416，EGFR和MAPK的高葡萄糖刺激磷酸化。这些抑制剂和siRNA以及SAPI敲除的Src敲除也废除了高葡萄糖诱导的这些靶标的磷酸化和胶原IV的积累。在STZ糖尿病小鼠中，白蛋白尿，Src pTyr-416升高，TACE活化，ERK和EGFR磷酸化，肾小球胶原蛋白积聚和足细胞丢失均被PP2抑制。这些数据表明Src在高葡萄糖-Src-TACE-肝素结合表皮生长因子-EGFR-MAPK信号通路中积累胶原蛋白的作用。因此，Src可以为糖尿病性肾病提供新的治疗靶标。

BACKGROUND & AIMS: :EGF receptor (EGFR) endocytosis is induced by stress in a manner dependent on the p38 MAPK family. Ligand and stresses such as X-rays, reportedly promote nuclear trafficking of endocytosed EGFR for regulation of gene transcription and DNA repair. We fail to detect EGFR endocytosis or nuclear transport following X-ray treatment of HeLa or head and neck cancer cells, despite extensive DNA damage induction. Apparent nuclear staining with EGFR extracellular domain antibody remained present despite reduced/absent EGFR expression, and so did not represent nuclear EGFR. UVB and UVC, but not X-ray or UVA, treatment induced p38 activation and EGFR endocytosis, although all of these stresses induced DNA damage, indicating that DNA damage alone is not sufficient to induce EGFR endocytosis. Increased reactive oxygen species (ROS) levels following UVB treatment, compared to that seen with X-rays, do not alone explain differences in p38 activation. UVB, like UVC, induced EGFR accumulation predominantly in perinuclear endosomes, rather than in the nucleus. Our morphological techniques identifying major changes in receptor distribution do not exclude the possibility that small but biologically relevant amounts of EGFR enter the nucleus. This study highlights the importance and limitations of morphological analyses of receptor distribution in understanding signaling outcome.

背景与目标： ：EGF受体（EGFR）的内吞作用是由应激以依赖于p38 MAPK家族的方式诱导的。据报道，配体和压力（例如X射线）可促进内吞EGFR的核转运，从而调节基因转录和DNA修复。尽管广泛的DNA损伤诱导作用，但在X射线治疗HeLa或头颈部癌细胞后，我们仍未检测到EGFR的内吞作用或核转运。尽管EGFR表达减少/缺失，但仍存在EGFR胞外域抗体的明显核染色，因此并不代表核EGFR。 UVB和UVC处理可诱导p38活化和EGFR内吞作用，而不是X射线或UVA，尽管所有这些压力均可诱导DNA损伤，表明仅DNA损伤不足以诱导EGFR内吞作用。与X射线相比，UVB处理后增加的活性氧（ROS）水平不能单独解释p38活化的差异。与UVC一样，UVB主要诱导EGFR聚集在核周内体中，而不是在细胞核中。我们的鉴定受体分布主要变化的形态学技术并不排除少量但生物学上相关的EGFR进入细胞核的可能性。这项研究强调了受体分布形态分析在理解信号转导结果中的重要性和局限性。

BACKGROUND & AIMS: BACKGROUND:Beyond progression after tyrosine kinase inhibitor in EGFR-positive non-small-cell lung cancer patients (BE-POSITIVE) was the first Italian multicenter observational study that reported the outcomes of first-generation epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitors (TKIs) in a "real-life" Caucasian EGFR-mutated non-small-cell lung cancer (NSCLC) population. The sharing of multi-institutional experiences represents a crucial strategy to enrich knowledge about uncommon EGFR mutations. Therefore, we performed a post hoc analysis of the BE-POSITIVE study. PATIENTS AND METHODS:Data of advanced NSCLC patients with uncommon EGFR mutations who received first-line first-generation EGFR-TKIs in 24 Italian Hospitals were collected. In this analysis we aimed to evaluate overall survival (OS), progression-free survival (PFS), and overall response rate (ORR) of EGFR-TKIs in NSCLC patients harboring uncommon EGFR mutations. RESULTS:Thirty-five patients harboring uncommon EGFR mutations (any mutation other than deletion 19 or substitution of leucine by arginine at codon 858) were included of the original 312 EGFR-mutated cases. Most of them were female (n = 20, 57.1%), former smokers (n = 23, 65.7%), with adenocarcinoma (n = 31, 88.6%). The most frequent EGFR mutations were G719X (n = 6, 17.2%) and L861Q (n = 5, 14.2%). The population presented an ORR of 25.7%, a median PFS of 5.19 months, and a median OS of 14.49 months. When stratified according to type of EGFR mutation, median OS ranged from 3.65 months for unspecified mutations to 21.29 for double EGFR mutations. Median PFS ranged from 1.77 months for unspecified mutations to 20.83 months for concomitant EGFR-anaplastic lymphoma kinase alteration. ORR varied from 0% in exon 18, 20 and double gene alteration to 66.6% in exon 19. CONCLUSION:Our study supports the existence of a strong outcome heterogeneity within patients harboring uncommon EGFR mutations, which needs to be clarified to achieve a real personalized treatment strategy.

背景与目标： 背景：酪氨酸激酶抑制剂在EGFR阳性非小细胞肺癌患者中取得的突破性进展（BE-POSITIVE）是第一项意大利多中心观察性研究，该研究报告了第一代表皮生长因子受体（EGFR）-酪氨酸激酶的结果现实生活中的白种人EGFR突变的非小细胞肺癌（NSCLC）人群中存在多种抑制剂（TKIs）。共享多机构经验代表了一项重要策略，可以丰富有关罕见的EGFR突变的知识。因此，我们对正研究进行了事后分析。
患者和方法：收集了意大利24所医院接受一线第一代EGFR-TKI的EGFR突变罕见的晚期NSCLC患者的数据。在这项分析中，我们旨在评估具有罕见EGFR突变的NSCLC患者的EGFR-TKIs的总生存期（OS），无进展生存期（PFS）和总缓解率（ORR）。
结果：35例罕见的EGFR突变病例包括了不常见的EGFR突变（除19号缺失或亮氨酸在858位密码子处亮氨酸替代以外的任何突变）。其中大多数是女性（n = 20，57.1％），曾吸烟者（n = 23，65.7％），患有腺癌（n = 31，88.6％）。最常见的EGFR突变是G719X（n = 6，17.2％）和L861Q（n = 5，14.2％）。人群的ORR为25.7％，PFS中位数为5.19个月，OS中位数为14.49个月。根据EGFR突变的类型进行分层时，中位OS​​范围从3.65个月（未指定突变）到21.29倍（双重EGFR突变）不等。 PFS的中位值范围从未指定突变的1.77个月到伴随的EGFR-再生障碍性淋巴瘤激酶改变的20.83个月不等。 ORR从外显子18、20和双基因改变的0％到外显子19的66.6％不等。
结论：我们的研究支持携带罕见EGFR突变的患者存在强大的预后异质性，需要阐明这一点以实现真正的个性化治疗策略。

BACKGROUND & AIMS: :Osteosarcoma is a primary malignancy of bone that affects children and adults. Here, we present the largest sequencing study of osteosarcoma to date, comprising 112 childhood and adult tumours encompassing all major histological subtypes. A key finding of our study is the identification of mutations in insulin-like growth factor (IGF) signalling genes in 8/112 (7%) of cases. We validate this observation using fluorescence in situ hybridization (FISH) in an additional 87 osteosarcomas, with IGF1 receptor (IGF1R) amplification observed in 14% of tumours. These findings may inform patient selection in future trials of IGF1R inhibitors in osteosarcoma. Analysing patterns of mutation, we identify distinct rearrangement profiles including a process characterized by chromothripsis and amplification. This process operates recurrently at discrete genomic regions and generates driver mutations. It may represent an age-independent mutational mechanism that contributes to the development of osteosarcoma in children and adults alike.

背景与目标： ：骨肉瘤是影响儿童和成人的原发性骨恶性肿瘤。在这里，我们介绍了迄今为止最大的骨肉瘤测序研究，包括112种儿童和成人肿瘤，涵盖了所有主要的组织学亚型。我们研究的关键发现是在8/112（7％）的病例中鉴定胰岛素样生长因子（IGF）信号基因的突变。我们在另外87个骨肉瘤中使用荧光原位杂交（FISH）验证了这一观察结果，并在14％的肿瘤中观察到了IGF1受体（IGF1R）扩增。这些发现可能会为将来在骨肉瘤中使用IGF1R抑制剂的患者选择药物提供依据。分析突变的模式，我们确定了独特的重排图谱，包括以色杆菌病和扩增为特征的过程。该过程在离散的基因组区域反复进行，并产生驱动突变。它可能代表了与年龄无关的突变机制，有助于儿童和成年人中骨肉瘤的发展。

BACKGROUND & AIMS: :The human epidermal growth factor receptor (HER) system is an intricately regulated system that plays critical roles in development and tumorigenesis. Here, we apply integrated experimentation and modeling to analyze HER receptor activation in a panel of cell lines expressing endogenous levels of EGFR/HER1 and different levels of HER2. A mathematical model that includes the fundamental processes involved in receptor activation and trafficking was used to fit the experimental data, and values of the independent parameters for active receptor dimer formation affinities, trafficking rates and relative phosphorylation levels were estimated. Obtained parameter values quantitatively support the existing ideas on the effect of HER2 on EGFR dynamics, and enable us to predict the abundances of various phosphorylated receptor dimers in the cell lines.

背景与目标： ：人类表皮生长因子受体（HER）系统是一个复杂的调控系统，在发育和肿瘤发生中起着至关重要的作用。在这里，我们应用综合实验和建模来分析一组表达内源性EGFR / HER1和不同水平HER2的细胞系中的HER受体激活。使用包括参与受体激活和运输的基本过程的数学模型来拟合实验数据，并估计活性受体二聚体形成亲和力，运输速率和相对磷酸化水平的独立参数的值。获得的参数值定量支持关于HER2对EGFR动力学影响的现有观点，并使我们能够预测细胞系中各种磷酸化受体二聚体的丰度。

BACKGROUND & AIMS: :The congenital vomer defect (CVD) is a rare and still partially unknown condition. Only few cases have been reported in the international literature and the large majority of them appeared to be isolated. We report a case of CVD detected in a 7-year-old girl affected by ectodermal dysplasia clefting syndrome caused by a mutation of the TP63 gene.

背景与目标： ：先天性犁骨缺损（CVD）是一种罕见且仍部分未知的疾病。国际文献中仅报道了少数病例，其中大多数似乎是孤立的。我们报告了一个在7岁的女孩中检测到的CVD病例，该女孩受TP63基因突变引起的外胚层发育不良裂口综合征的影响。

BACKGROUND & AIMS: :The Saccharomyces cerevisiae TIM10 complex (TIM10c) is an ATP-independent chaperone of the mitochondrial intermembrane space, involved in transport of polytopic membrane proteins. The complex is an alpha(3)beta(3) hexamer of Tim9 and Tim10 subunits. We have generated specific mutations in charged residues in the central core domain of each subunit delineated by the characteristic twin CX(3)C motif, and investigated the effect of these mutations on subunit folding, complex assembly and TIM10 function in vitro and in vivo. Any combination of mutations that included a specific glutamate residue, conserved in all known Tim9 and Tim10 sequences, abolished assembly of the TIM10 complex. In vivo complementation analyses using a MET3-TIM10 strain that is selectively inactivated for the expression of wild-type Tim10 showed that (i) an N-terminal deleted version of Tim10 that was previously shown to be defective in substrate binding is lethal under all conditions, but (ii) the charged residues mutant of Tim10 that is defective in assembly with Tim9 can restore growth in glucose, but not in non-fermentable carbon sources. These data suggest that formation of the hexamer is beneficial but not vital for TIM10 function, whilst the N-terminal substrate-binding region of Tim10 is essential in vivo.

背景与目标： ：酿酒酵母TIM10复合物（TIM10c）是线粒体膜间空间的一个不依赖ATP的分子伴侣，参与多膜蛋白的转运。该复合物是Tim9和Tim10亚基的alpha（3）beta（3）六聚体。我们已经在每个亚基的中央核心域带电残基中生成特定的突变，这些突变由特征性双胞胎CX（3）C图案描绘，并研究了这些突变对亚基折叠，复杂装配和TIM10功能的影响。在所有已知的Tim9和Tim10序列中均保守的，包括特定谷氨酸残基的任何突变组合都可消除TIM10复合体的装配。使用针对野生型Tim10的表达而选择性失活的MET3-TIM10菌株的体内互补分析显示（i）先前被证明在底物结合方面有缺陷的Tim10的N末端缺失版本在所有情况下都是致死性的，但（ii）与Tim9组装有缺陷的Tim10的带电荷残基突变体可以恢复葡萄糖的生长，但不能恢复不可发酵的碳源。这些数据表明六聚体的形成对TIM10功能是有益的，但不是至关重要的，而Tim10的N端底物结合区在体内则是必不可少的。

BACKGROUND & AIMS: :Seven fulvestrant resistant cell lines derived from the estrogen receptor alpha positive MCF-7 human breast cancer cell line were used to investigate the importance of epidermal growth factor receptor (ErbB1-4) signaling. We found an increase in mRNA expression of EGFR and the ErbB3/ErbB4 ligand heregulin2 (hrg2) and a decrease of ErbB4 in all resistant cell lines. Western analyses confirmed the upregulation of EGFR and hrg2 and the downregulation of ErbB4. Elevated activation of EGFR and ErbB3 was seen in all resistant cell lines and the ErbB3 activation occurred by an autocrine mechanism. ErbB4 activation was observed only in the parental MCF-7 cells. The downstream kinases pAkt and pErk were increased in five of seven and in all seven resistant cell lines, respectively. Treatment with the EGFR inhibitor gefitinib preferentially inhibited growth and reduced the S phase fraction in the resistant cell lines concomitant with inhibition of Erk and unaltered Akt activation. In concert, inhibition of Erk with U0126 preferentially reduced growth of resistant cell lines. Treatment with ErbB3 neutralizing antibodies inhibited ErbB3 activation and resulted in a modest but statistically significant growth inhibition of two resistant cell lines. These data indicate that ligand activated ErbB3 and EGFR, and Erk signaling play important roles in fulvestrant resistant cell growth. Furthermore, the decreased level of ErbB4 in resistant cells may facilitate heterodimerization of ErbB3 with EGFR and ErbB2. Our data support that a concerted action against EGFR, ErbB2 and ErbB3 may be required to obtain complete growth suppression of fulvestrant resistant cells.

背景与目标： ：使用七种来自雌激素受体α阳性MCF-7人乳腺癌细胞的抗氟维司群抗性细胞系，调查表皮生长因子受体（ErbB1-4）信号传导的重要性。我们发现在所有耐药细胞系中EGFR和ErbB3 / ErbB4配体heregulin2（hrg2）的mRNA表达增加，而ErbB4的减少。 Western分析证实了EGFR和hrg2的上调以及ErbB4的下调。在所有耐药细胞系中均观察到EGFR和ErbB3的激活增强，并且ErbB3的激活是通过自分泌机制发生的。仅在亲本MCF-7细胞中观察到ErbB4激活。下游激酶pAkt和pErk在七个抗性细胞系中的五个中和在所有七个抗性细胞系中分别增加。用EGFR抑制剂吉非替尼治疗可优先抑制生长并降低耐药细胞系中的S期分数，同时抑制Erk和未改变的Akt激活。一致地，用U0126抑制Erk优先降低了抗性细胞系的生长。用ErbB3中和抗体处理可抑制ErbB3活化，并导致对两种抗性细胞系的抑制作用达到中等但统计学上显着的增长。这些数据表明配体激活的ErbB3和EGFR，以及Erk信号传导在耐氟司韦特的细胞生长中起重要作用。此外，耐药细胞中ErbB4水平降低可能有助于ErbB3与EGFR和ErbB2异源二聚化。我们的数据支持可能需要针对EGFR，ErbB2和ErbB3的协同作用才能获得对氟维司群抗性细胞的完全生长抑制。