Decrease in multiple sclerosis with acute transverse myelitis in Japan.

Acute transverse myelitis (ATM) may be a manifestation of multiple sclerosis (MS) and was reported to be more common among Japanese MS patients than in Caucasian MS patients. Recently there are arguments whether clinical manifestations of MS may have changed. Therefore, we studied the frequency of ATM in MS and the clinical subtypes of MS in 86 clinically definite MS patients whose onsets were in 1970-1979, 1980-1989, and 1990-1998 in Sendai City, Japan. Fifty-six of the patients were women and 30 were men. Forty-four patients had the conventional form of MS (C-MS) commonly seen in Western countries, and 42 had optic-spinal or spinal forms of MS (OSS-MS). Twenty MS patients had ATM, and all of them were belong to optic-spinal form of MS. ATM was not seen in any cases of C-MS. The mean onset age (years) of the clinical subtypes was 25.5 in C-MS, 34.1 in OSS-MS without ATM, and 30.9 in OSS-MS with ATM. Among the patients whose onset of the disease was in 1970-1979, 60.0% of them were cases of OSS-MS with ATM, but such cases were markedly decreased to 5.3% in 1990-1998. In contrast, the frequency of C-MS increased to 63.2% in 1990-1998 compared with 20.0% in 1970-1979. Analysis of the data by the year of birth of the patients showed similar results. Our data suggest that the frequency of ATM in MS markedly decreased, and that of C-MS increased during the last 30 years in Sendai, Japan. Since the genetic background of Japanese has not changed, some exogenous factors, such as food, infectious microorganisms, and chemicals in our environment, may be responsible for the change.     

肿瘤与结核标志蛋白整合技术对卵巢恶性肿瘤患者预后的判定作用

目的分析卵巢恶性肿瘤患者血清CA125水平和SA/ATM二联标志蛋白表达信息,探讨双联整合技术用于临床康复鉴别诊断的临床价值。方法经核磁共振氢谱NMR1H仪捕获49例结核性腹膜炎ATM-Pr谱学特征,用生化显色法和磁分离酶免疫法检测101例卵巢恶性肿瘤,985例其他妇科疾患血清标志蛋白,进行对比分析。结果血清CA125和SA水平在卵巢恶性肿瘤和结核性腹膜炎组均明显升高(差异无显著性,P>0.05);>子宫颈癌>子宫肌瘤、卵巢良性肿瘤、单纯性卵巢囊肿、盆腔炎性包块;血清ATM-Pr谱学特征显示:卵巢恶性肿瘤和结核性腹膜炎在信息特征峰表达上有明显差异(P<0.01)。结论SA和CA125在卵巢恶性肿瘤诊断中有较高准确性,但特异性受结核疾病干扰。SA/ATM双联检测可有效用于肿瘤/结核的筛查和鉴别,有较好的特异性,方法快捷更具早发现适用性,并可作为临床康复判定的指标之一。     

抗凝血酶基因C2759T(Leu99Phe)突变导致抗凝血酶缺陷症的分子机制研究

目的研究抗凝血酶(AT)基因C2759T(Leu99Phe)突变引起AT缺陷症的分子机制。方法用大引物法构建C2759T突变体AT重组表达质粒(ATM2759)，并将其和正常AT重组质粒(ATN)分别用Superfect试剂转染COS7细胞或CHO细胞，进行体外表达试验和细胞免疫荧光染色。、结果转染ATM2759的COS7细胞，培养上清液和细胞裂解液中的AT抗原(AT：Ag)分别为ATN的35．63％和174．97％，而培养上清液中的AT活性(AT：A)为ATN的47．73％。细胞免疫荧光试验证实，转染ATM2759的CHO细胞，其荧光强度明显高于转染ATN的CHO细胞。结论Leu99Phe 突变可能不是由于影响AT与肝素的结合而导致AT缺陷，而是由于突变导致AT分泌障碍和细胞内滞留所致。     

ATM基因单核苷酸多态性与食管癌易感性的研究

目的食管癌是世界上最常见的恶性肿瘤之一;毛细血管扩张性共济失调症突变基因(ATM)是导致毛细血管扩张性共济失调症发生的致病基因,与肿瘤的发生密切相关;本研究旨在探讨ATM基因单核苷酸多态性与食管癌发生的相关性。方法收集2009年9月至2010年12月期间江苏省淮安市楚州医院就诊的135例食管癌患者标本以及135例健康体检者作为健康对照组。ATM基因单核苷酸多态性分型检测采用Taqman技术,应用Logistc回归统计分析ATM单核苷酸多态性与食管癌发生的相关性。结果吸烟者食管癌的发生率明显高于不吸烟者,差异有统计学意义(χ2=10.252,P=0.001,P<0.05);有肿瘤家族史者患食管癌的概率明显高于没有肿瘤家族史者,差异有统计学意义(χ2=12.199,P=0.000,P<0.05);在食管癌患者组中,A/A基因型占31.9%,A/G基因型占38.5%,G/G基因型占29.6%;而健康对照组中相应的数值分别为25.9%、48.9%和25.2%。以A/A基因型相比,A/G基因型的OR值为0.798(95%CI:0.418-1.521,P=0.493);G/G基因型的OR值为0.534(95%CI:0.265-1.079,P=0.080)。结论 ATM单核苷酸多态性可能与淮安地区的食管癌有关,为食管癌的保护性因素。     

急性横贯性脊髓炎预后影响因素的研究进展

急性横贯性脊髓炎(acute transverse myelitis,ATM)病因不明,目前多认为与感染后引起的自身免疫反应相关,部分患者于疫苗接种后发病。脊髓炎可引起脊髓水肿变软,导致病变平面以下运动、感觉等功能障碍。ATM患者的预后不一,从完全康复到致残,甚至死亡。大约1/3的患者恢复良好,1/3的患者恢复尚可,剩下1/3的患者病情很少或基本没有好转。ATM的预后取决于病变的严重程度及合并症的情况,但更精确的预测是困难的。     

p53和ATM基因缺失在慢性淋巴细胞白血病患者中的预后价值

目的 探讨p53和ATM基因缺失在慢性淋巴细胞白血病患者(CLL)中的预后价值.方法 采用间期荧光原位杂交(FISH)技术和p53、ATM基因的序列特异性DNA探针对80例CLL患者的染色体标本进行p53和ATM基因缺失的检测,分析p53和ATM基因缺失与外周血淋巴细胞绝对计数、Binet分期、血清乳酸脱氢酶(LDH)水平、β2微球蛋白(β2-MG)水平、ZAP-70表达、CD38表达、含氟达拉滨治疗疗效之间的相关性,单因素生存分析采用Kaplan-Meier法绘制生存曲线和Log-rank检验,多因素生存分析采用COX多元回归分析.结果 80例CLL患者中14例(17.5%)伴有p53基因缺失,9例(11.3%)伴有ATM基因缺失,其中3例(3.8%)同时伴有p53和ATM基因缺失.p53和ATM基因缺失与患者性别、年龄、Binet分期、外周血淋巴细胞绝对计数、血清LDH及β2-MG、ZAP-70表达水平无明显相关性;CD38高表达组中,p53和ATM基因缺失的发生率均明显高于CD38低表达组,且差异有统计学意义(P=0.025和P=0.001).41例患者接受含氟达拉滨方案治疗,32例不伴有p53和(或)ATM基因缺失的患者中,12例(37.5%)获得完全缓解(CR),9例伴有p53和(或)ATM基因缺失的患者均未获得CR(P=0.047).单因素生存分析显示,伴有p53基因缺失组的生存期明显较不伴有缺失组短,经Log-rank检验差异有统计学意义(P＜0.01);而伴有与不伴有ATM基因缺失组的生存期差异无统计学意义(P=0.556).COX多元回归分析显示,p53基因缺失(P:0.014)和CD38表达水平(P=0.017)是判断CLL预后的独立因素.结论 伴有p53和(或)ATM基因缺失的患者,接受含氟达拉滨方案治疗疗效差,FISH检测p53和ATM基因缺失在CLL预后判断和指导临床治疗中具有重要价值.     

Antisense ATM gene therapy: a strategy to increase the radiosensitivity of human tumors

Atm, the gene mutated in ataxia-telangiectasia (AT) patients, is an essential component of the signal transduction pathway that responds to DNA damage due to ionizing radiation (IR). We attenuated ATM protein expression in human glioblastoma cells by expressing antisense RNA to a functional domain of the atm gene. While ATM expression decreased, constitutive expression of p53 and p21 increased. Irradiated ATM-attenuated cells failed to induce p53, demonstrated radioresistant DNA synthesis, and increased radiosensitivity. Antisense-ATM gene therapy in conjunction with radiation therapy may provide a novel strategy for the treatment of cancer.     

间期荧光原位杂交技术检测慢性淋巴细胞白血病ATM基因缺失

目的探讨慢性淋巴细胞白血病(CLL)中ATM基因缺失及其与其他染色体异常及临床分期的相关性。方法运用间期荧光原位杂交技术(I-FISH)和Spectrum OrangeTM标记的位于11q22.3的序列特异性DNA探针ATM对50例初诊CLL患者的染色体标本进行了ATM缺失的检测,同时检测del(13q14)、del(17p13.1)和免疫球蛋白重链基因重排。临床分期按照Binet分期方法。结果50例患者中有6例(12%)ATM缺失;其中4例伴有其它染色体异常。20例Binet A期患者中,3例(15%)存在异常;10例Binet B期患者中,2例(20%)存在异常;13例Binet C期患者中,1例(7.7%)存在异常。ATM缺失在Binet A、B及C期中无统计学差异(P>0.05)。结论I-FISH与常规染色体分析技术相比是一种快速、准确及敏感的方法,对我国CLL患者的预后预测价值有待进一步的深入研究。     

不育男性沙眼衣原体感染与血清五种自身抗体检测及分析

目的：探讨沙眼衣原体（CT）感染与血清中抗精子抗体（AsAb)、抗心磷脂抗体（ACA）、抗核抗体（ANA）、抗甲状腺球蛋白抗体（ATG）、抗甲状腺微粒体抗体（ATM）之间的关系。方法；用ELISA法、放射免疫分析法和间接免疫荧光法分别检测50例正常生育男性（正常对照组）和108例CT感染不育男性（CT感染组）的血清中AsAb、ACA、ANA、ATG、ATM。结果：CT感染组AsAb检出率明显高于对照组，具有显著性差异（P＜0．01），ACA次之（P＜0．05），而ANA、ATG、ATM两组间无显著性差异（P＞0．05）。结论：生殖道CT感染可引起机体免疫反应而产生AsAb，是其导致男性不育的重要原因之一。     

Functional expression of ATM gene carried by HSV amplicon vector in vitro and in vivo

Ataxia-telangiectasia (AT) is a human autosomal recessive disease with a pleiotropic phenotype characterized by cerebellar degeneration, immunodeficiency, premature aging, cancer predisposition, and radiation sensitivity. The gene mutated in AT, ATM (for AT-mutated), had been cloned and found to have ionizing radiation and oxidative stress-inducible kinase activity. No treatment can stop the progression of the disease. In this study, the complete open-reading frame of ATM cDNA was cloned into a Herpes simplex virus type-1 (HSV-1) amplicon vector (pTO-ATM), and the transduction of cultured AT cells was demonstrated by immunohistochemistry and Western blot analysis. Functional gene expression was evaluated by cell colony-forming assays following exposure to oxidative stress. The survival of AT cells with ATM gene transduction was about 100% higher compared to nontransduced cells after t-butyl hydroperoxide treatments. Next, the normal ATM gene expression in different regions of the rat brain was studied. Immunohistochemistry staining demonstrated weak endogenous ATM protein expression in neurons of the caudate-putamen, with significantly higher levels of expression detected in neurons in other brain regions. Exogenous ATM gene expression from pTO-ATM after viral transduction in the caudate-putamen of the adult rat was examined. At 3 days after injection of the pTO-ATM viral vector, abundant positive ATM staining of the neurons was found at the injection sites, in comparison to the controls. These data demonstrate that the relatively large ATM cDNA can be transduced and expressed in vitro and in vivo from an HSV amplicon viral vector. These data provide initial evidence that the replacement of the ATM gene into the cells of AT patients might be possible some day.     

K562和SiHA细胞株经γ-射线照射后细胞周期阻滞与ATM表达量的关系研究

共济失调毛细血管扩张症（ataxia telangiectasis,A-T）是由ATM（ataxia telangiectasis mutant）基因突变所致,其突出特点是对放射线敏感.为探讨K562和SiHA两种肿瘤细胞株ATM表达量与γ-射线照射后细胞周期阻滞即自我保护功能之间的关系,应用半定量RT-PCR测量它们的ATM mRNA表达,同时以6、10和15 Gy 60Co γ射线分别照射细胞,并于照射后6、12、24、48及60小时观察细胞周期阻滞现象和凋亡率的变化.结果显示,K562细胞株ATM RNA相对表达量为0.04,而在SiHA细胞株为0.80,SiHA的ATM RNA表达量约为K562的20倍.结论：照射后K562和SiHA细胞株均表现G2/M期阻滞,K562细胞周期阻滞即自我保护机制明显比SiHA差.     

Lengthening the hamstring muscles without stretching using "awareness through movement"

BACKGROUND AND PURPOSE: Passive stretching is widely used to increase muscle flexibility, but it has been shown that this process does not produce long-term changes in the viscoelastic properties of muscle as originally thought. The authors tested a method of lengthening hamstring muscles called "Awareness Through Movement" (ATM) that does not use passive stretching. SUBJECTS: Thirty-three subjects who were randomly assigned to ATM and control groups met the screening criteria and completed the intervention phase of the study. METHODS: The ATM group went through a process of learning complex active movements designed to increase length in the hamstring muscles. Hamstring muscle length was measured before and after intervention using the Active Knee Extension Test. RESULTS: The ATM group gained significantly more hamstring muscle length (+7.04 degrees ) compared with the control group (+1.15 degrees ). DISCUSSION AND CONCLUSION: The results suggest that muscle length can be increased through a process of active movement that does not involve stretching. Further research is needed to investigate this finding.     

Fanconi anemia pathway-deficient tumor cells are hypersensitive to inhibition of ataxia telangiectasia mutated

The Fanconi anemia (FA) pathway maintains genomic stability in replicating cells. Some sporadic breast, ovarian, pancreatic, and hematological tumors are deficient in FA pathway function, resulting in sensitivity to DNA-damaging agents. FA pathway dysfunction in these tumors may result in hyperdependence on alternative DNA repair pathways that could be targeted as a treatment strategy. We used a high-throughput siRNA screening approach that identified ataxia telangiectasia mutated (ATM) as a critical kinase for FA pathway–deficient human fibroblasts. Human fibroblasts and murine embryonic fibroblasts deficient for the FA pathway were observed to have constitutive ATM activation and Fancg^–/–Atm^–/– mice were found to be nonviable. Abrogation of ATM function in FA pathway–deficient cells resulted in DNA breakage, cell cycle arrest, and apoptotic cell death. Moreover, Fanconi anemia complementation group G– (FANCG-) and FANCC-deficient pancreatic tumor lines were more sensitive to the ATM inhibitor KU-55933 than isogenic corrected lines. These data suggest that ATM and FA genes function in parallel and compensatory roles to maintain genomic integrity and cell viability. Pharmaceutical inhibition of ATM may have a role in the treatment of FA pathway–deficient human cancers.     

p38MAPK对肿瘤细胞ATM、ATR和p53信号通路的影响

目的 研究p38MAPK对细胞周期信号转导通路中重要蛋白激酶ATM、ATR、P53的影响并探讨其可能的机制.方法 采用人类结肠癌细胞株HT-29和肺癌细胞株SPAC-1进行研究,分别使用紫外线（UV）刺激和p38MAPK特异性抑制剂SB203580抑制肿瘤细胞中的p38MAPK,通过Western blot技术检测ATM、ATR、P53蛋白磷酸化情况,同时用real-time PCR技术检测细胞中相应基因的表达情况.结果 紫外线能上调ATM、ATR、p53和p38MAPK基因的表达,使用p38MAPK特异抑制剂SB203580能够使ATM、ATR和p53的表达降低.结论 P38MAPK可以调控ATM、ATR和p53的表达,ATM-Chk2信号通路和p38MAPK信号通路之间可能存在交互联系.     

Ataxia telangiectasia

Ataxia-telangiectasia(AT), an autosomal recessive disorder characterized by cerebellar degeneration, immunodeficiency, cancer predisposition and radiation sensitivity, is caused by mutations in a gene named ATM(AT, mutated), which encodes a 370 kDa serine-threonine kinase, whose catalytic domain is structurally related to the catalytic subunit of phosphatidylinositol 3-kinase(PI3K). ATM has been recently revealed to be involved in DNA damage recognition and cell cycle control in response to ionizing radiation damage. Further investigations of the multiple roles of ATM will explain other disease features, such as cerebellar degeneration in ATM in the near future. This review summarizes some of the recent research developments in ATM functions and their relationship to the clinical phenotypes of AT.     

ATM regulates ATR chromatin loading in response to DNA double-strand breaks

DNA double-strand breaks (DSBs) are among the most deleterious lesions that can challenge genomic integrity. Concomitant to the repair of the breaks, a rapid signaling cascade must be coordinated at the lesion site that leads to the activation of cell cycle checkpoints and/or apoptosis. In this context, ataxia telangiectasia mutated (ATM) and ATM and Rad-3-related (ATR) protein kinases are the earliest signaling molecules that are known to initiate the transduction cascade at damage sites. The current model places ATM and ATR in separate molecular routes that orchestrate distinct pathways of the checkpoint responses. Whereas ATM signals DSBs arising from ionizing radiation (IR) through a Chk2-dependent pathway, ATR is activated in a variety of replication-linked DSBs and leads to activation of the checkpoints in a Chk1 kinase-dependent manner. However, activation of the G2/M checkpoint in response to IR escapes this accepted paradigm because it is dependent on both ATM and ATR but independent of Chk2. Our data provides an explanation for this observation and places ATM activity upstream of ATR recruitment to IR-damaged chromatin. These data provide experimental evidence of an active cross talk between ATM and ATR signaling pathways in response to DNA damage.     

Immunoassay to measure ataxia-telangiectasia mutated protein in cellular lysates

BACKGROUND: Ataxia-telangiectasia (A-T) is a neurologic disorder caused by mutations in the ataxia-telangiectasia mutated (ATM) gene. A clinical diagnosis of A-T is confirmed by radiosensitivity testing and immunoblotting for ATM protein. Because both of these tests have long turnaround times (> or =3 months), we developed a rapid immunoassay to measure ATM protein and determined its sensitivity and specificity for diagnosing A-T. METHODS: Recombinant ATM protein was used for standardization. Lysates of lymphoblastoid cell lines (LCLs) and peripheral blood mononuclear cells (PBMCs) from A-T patients, controls, and A-T heterozygotes were tested for ATM protein by immunoassay. RESULTS: Between-run imprecision (CV) was < or =13%. Nuclear lysates from control LCLs and PBMCs had ATM protein concentrations of 49-610 microg/L and 48-943 microg/L, respectively. ATM protein was not detectable in LCL nuclear lysates from 18 of 21 A-T patients. The three remaining A-T patients had trace amounts of ATM protein, which was confirmed on immuoblots. ATM protein was also detectable in whole-cell lysates from 4 x 10(6) cells at concentrations of 64-463 microg/L and 42-444 microg/L for control LCLs and PBMCs, respectively. A-T heterozygotes had ATM protein concentrations of 52-98 microg/L. ATM protein was stable in PBMCs stored for 1 month at -70 degrees C, but rapidly decreased after 1 day in unprocessed blood. CONCLUSIONS: This ATM protein immunoassay can be used to confirm a diagnosis of A-T in 2 days on small numbers of PBMCs and can potentially identify A-T carriers and individuals at increased risk for cancer.     

Topoisomerase poisons differentially activate DNA damage checkpoints through ataxia-telangiectasia mutated-dependent and -independent mechanisms

Camptothecin and Adriamycin are clinically important inhibitors for topoisomerase (Topo) I and Topo II, respectively. The ataxia-telangiectasia mutated (ATM) product is essential for ionizing radiation-induced DNA damage responses, but the role of ATM in Topo poisons-induced checkpoints remains unresolved. We found that distinct mechanisms are involved in the activation of different cell cycle checkpoints at different concentrations of Adriamycin and camptothecin. Adriamycin promotes the G(1) checkpoint through activation of the p53-p21(CIP1/WAF1) pathway and decrease of pRb phosphorylation. Phosphorylation of p53(Ser20) after Adriamycin treatment is ATM dependent, but is not required for the full activation of p53. The G(1) checkpoint is dependent on ATM at low doses but not at high doses of Adriamycin. In contrast, the Adriamycin-induced G(2) checkpoint is independent on ATM but sensitive to caffeine. Adriamycin inhibits histone H3(Ser10) phosphorylation through inhibitory phosphorylation of CDC2 at low doses and down-regulation of cyclin B1 at high doses. The camptothecin-induced intra-S checkpoint is partially dependent on ATM, and is associated with inhibitory phosphorylation of cyclin-dependent kinase 2 and reduction of BrdUrd incorporation after mid-S phase. Finally, apoptosis associated with high doses of Adriamycin or camptothecin is not influenced by the absence of ATM. These data indicate that the involvement of ATM following treatment with Topo poisons differs extensively with dosage and for different cell cycle checkpoints.