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91.
Itoh K Naganawa Y Matsuzawa F Aikawa S Doi H Sasagasako N Yamada T Kira J Kobayashi T Pshezhetsky AV Sakuraba H 《Journal of human genetics》2002,47(1):29-37
Three novel missense mutations in the human lysosomal sialidase gene causing amino acid substitutions (P80L, W240R, and P316S)
in the coding region were identified in two Japanese sialidosis patients. One patient with a severe, congenital form of type
2 sialidosis was a compound heterozygote for 239C-to-T (P80L) and 718T-to-C (W240R). The other patient with a mild juvenile-onset
phenotype (type 1) was a homozygote for the base substitution of 946C-to-T (P316S). None of these mutant cDNA products showed
enzymatic activity toward an artificial substrate when coexpressed in galactosialidosis fibroblastic cells together with protective
protein/cathepsin A (PPCA). All mutants showed a reticular immunofluorescence distribution when coexpressed with the PPCA gene in COS-1 cells, suggesting that the gene products were retained in the endoplasmic reticulum/Golgi area or rapidly degraded
in the lysosomes. Homology modeling of the structural changes introduced by the mutations predicted that the P80L and P316S
transversions cause large conformational changes including the active site residues responsible for binding the sialic acid
carboxylate group. The W240R substitution was deduced to influence the molecular surface structure of a limited region of
the constructed models, which was also influenced by previously identified V217M and G243R transversions.
Received: Stptember 21, 2001 / Accepted: November 2, 2001 相似文献
92.
Two novel mutations of the β-hexosaminidase α subunit gene were identified in Japanese patients with the infantile form of
Tay-Sachs disease. One mutation was a one-base deletion at nt613C, which generated a stop codon at two codons downstream,
in three unrelated patients. The other mutation was a one-base substitution of G-to-A at IVS 5, +1, which caused a splicing
abnormality, in one patient. A missense mutation of R170W, which has already been reported in other ethnic groups, was also
newly identified in one patient. In 1993, the most common mutation (IVS 5, −1G → T) in Japanese patients with Tay-Sachs disease
was reported as the major mutation in Japan accounting for 80% of 56 mutant alleles from 28 unrelated patients. The deletion
of nt613C was the second most common mutation, accounting for 5% of the mutant alleles. The previously reported mutation IVS
5, −1G → T and the nt613C deletion found in this study together accounted for 85% of the mutations causing Tay-Sachs disease
among Japanese. Since these two mutations were located in or close to exon 6 and since they abolish Fok I (IVS 5, −1G → T) and Sfa NI (nt613C deletion) restriction sites, respectively, they were screened rapidly by single polymerase chain reaction followed
by digestion with these enzymes.
Received: November 10, 1998 / Accepted November 14, 1998 相似文献
93.
Jean-Aimé Simoneau Margit Kaufmann Karl-Thomas Härtner Dirk Pette 《Pflügers Archiv : European journal of physiology》1989,414(6):629-633
This study compares changes in contractile properties, Parvalbumin content, and Ca2+-uptake by the sarcoplasmic reticulum (SR) of low-frequency stimulated rat and rabbit tibialis anterior (TA) muscles. Time to peak tension increased 1.8-fold in 35-day stimulated rabbit TA, while no change occurred in rat TA. Isometric twitch tension increased 2-fold in rabbit TA, but was unaltered in rat TA. Parvalbumin (PA) content was more than 90% reduced in rabbit TA, but only 60% in rat TA after 35 days. At this time, PA content of the stimulated rat TA was still higher than that of normal rabbit TA. Taking into account the suggested role of PA as a cytosolic Ca2+ buffer, its decrease could lead to an impaired free Ca2+-decay with a prolonged active state and a higher tension output during a single twitch. This would explain why chronic stimulation led to an increase in isometric twitch tension in rabbit TA, but not in rat TA. The 1.6-fold rise in half-relaxation time of 35-day stimulated rat and rabbit TA most likely resulted from a 50% reduced Ca2+-uptake by the SR, due to a still unknown modification of the Ca2+-transport ATPase. 相似文献
94.
We report on clinical samples Stuttgart/97, Berlin/99 and Jasi/99 associated with aseptic meningitis. All three samples contained echovirus 4 (E4) but Stuttgart/97 was simultaneous infected with echovirus 30 (E30). The genetic relationship of the E4 strains was assessed using RT-PCR and direct sequencing of amplicons derived from the genomic region encoding the capsid protein VP1. The sequences have been compared with each other and with sequences of further E4 strains obtained from GenBank. The analysis confirms that sequences of recent isolates have drifted away from elderly strains over a longer period of time. Several amino acid changes in assumed antigenic sites of the VP1 gene may be sufficient to cause changes in antigenic specificity and therefore they may be a reason for failure of serological typing of some new antigenic E4 variants. 相似文献
95.
常见呼吸道病毒分子鉴别诊断技术的建立 总被引:3,自引:0,他引:3
目的 建立一种常见呼吸道感染病毒的快速检测方法,为尽早诊断、减少疾病的传播以及为临床提供良好的治疗依据.方法 采用液体芯片检测技术结合靶向多重RT-PCR技术建立可以同时检测13种呼吸道病原的检测技术.结果 该方法特异性方面检测13种常见呼吸道病毒没有交叉,标本的检出率为100%;灵敏度方面达到10e2-10e1(pfu/ml).结论 该分子鉴别诊断可应用常见呼吸道病毒的检测,协助诊断病毒引起的病毒性呼吸道感染. 相似文献
96.
一体化假肢是以聚合物为材料从接受腔到假腿一体成型的新型下肢假肢,它比传统型假肢更经济、美观、轻便,具有较大的应用前景。目前的相关研究主要集中在设计与制作及少量的临床研究方面。由于一体化假肢与传统型假肢在结构上的差异,有必要对其进行应力分析。本研究的目的是开展内骨架一体化假肢的生物力学研究,本研究基于内骨架一体化小腿假肢的真实几何构型,建立三维有限元模型,计算该模型在模拟Heel OH步态时相的载荷作用下的应力分布;在保持该模型的几何形状不变的情况下,建立了三个不同壁面厚度的一体化小腿假肢的有限元模型,分析壁面厚度对一体化小腿假肢应力分布的影响;通过分别赋予模型四种不同高分子聚合物的材料力学特性值,分析不同材料的一体化假肢的应力分布特点;分别对模型施加与正常步态的五个典型时相对应的载荷,分析一体化小腿假肢在各步态时相的应力分布特点。本研究结果对一体化假肢设计有指导价值。 相似文献
97.
dos Santos CN Rocha CF Cordeiro M Fragoso SP Rey F Deubel V Desprès P 《Virus research》2002,90(1-2):197-205
We have investigated the genetic diversity of dengue type-1 (DEN-1) virus in Brazil. The full nucleotide sequences of three DEN-1 virus isolated from DEN fever (DF) and DEN hemorrhagic fever patients in northeastern Brazil in 1997 (BR/97) and one from a DF patient in the south of Brazil in 2001 (BR/01) were compared to that of the reference strain BR/90 obtained in the city of Rio de Janeiro in 1990. Sequence analysis showed that the structural proteins were remarkably conserved between all isolates. A total of 27 amino acid changes occurred throughout the non-structural proteins. Among them, nine amino acid substitutions were specific of BR/97 and BR/01 isolates, indicating that in situ evolution of these strains had occurred. Within the BR/97 and BR/01 samples, some amino acid substitutions have been previously identified in DEN-1 virus strains sequenced so far, suggesting that recombination events might have occurred. 相似文献
98.
目的通过对上海地区腹泻住院患儿进行诺若病毒检测,对其流行株进行基因序列的测定,以了解诺若病毒在上海地区的流行特征,为该病原体所致腹泻的防治提供基础数据和理论依据。方法收集2001至2005年复旦大学附属儿科医院5岁以下腹泻住院患儿的粪便标本。首先进行轮状病毒的检测,在轮状病毒抗原阴性标本中,每隔8个标本按编号顺序行机械随机抽样,建立RT-PCR方法进行诺若病毒的检测。对PCR产物进行双向测序,测序结果通过Clustal W和Mega 4.1软件进行分析。结果研究期间共收集腹泻患儿粪便标本5534份,轮状病毒抗原阴性4084份,机械随机抽得484份用于诺若病毒检测,45/484份(9.3%)检测到诺若病毒。对诺若病毒感染季节分布和患儿年龄特点的分析表明,除4月和7月份未检测到诺若病毒外,其余各月份均检测到诺若病毒,其高发的月份是8至11月。5~6月也呈一个小高峰。77.8%(35/45)的患儿<2岁,其中6~11个月的患儿所占比例最高,达35.6%(16/45),<6个月的婴儿占20%(9/45)。GⅡ-4型是这5年间尤其是2003年之后的主要流行型别,2001至2002年尚存在其他的流行型别GⅡ-3和GⅡ-7... 相似文献
99.
采用柱上聚合的方法制备血红蛋白溶液,解决传统血红蛋白聚合过程中由于戊二醛的活性过高导致平均分子量大、产物分子量分布宽的问题。该方法利用阳离子交换剂对修饰度(聚合度)小的血红蛋白吸附能力大的原理,使其在柱上富积,同时加入戊二醛进行聚合反应。结果表明该方法能比较有效地缩小聚合血红蛋白的分子量分布。 相似文献
100.
Lipoblastoma is a relatively rare tumor that occurs in infancy and early childhood and arises from embryonic white fat. Although a benign tumor, lipoblastomas tend to recur and may resemble myxoid liposarcoma. The authors report 26 cases over a 15-year period at Texas Children's Hospital. There was a slight female predilection (14F:12M). The most common symptom was a painless mass with or without increasing size. The trunk, extremities, head and neck, retroperitoneum, inguinal canal, peritoneal cavity, and lung were the tumor sites. Most tumors were circumscribed lipoblastomas and the minority were diffuse infiltrative lipoblastomatosis. Reexcision for residual or recurrent tumor was necessary more frequently in patients with lipoblastomatosis. Histopathologic examination and ultrastructural examination revealed cellular neoplasms composed of immature adipocytes with relatively well-defined septa, frequent lipoblasts, a fine vascular network, and often a myxoid appearance resembling myxoid liposarcoma. Cytogenetics was performed in 4 cases with chromosome 8q abnormality being most common. The major concern with lipoblastoma in children is to completely excise the tumor to avoid leaving residual tumor and to prevent recurrences. Confusion with myxoid liposarcoma, well-differentiated liposarcoma, and typical lipomas may occur. Although asymptomatic, lipoblastomas may cause dysfunction of other organ systems due to mass effect. Complete surgical excision with at least 2 years of follow-up is the preferred therapy. 相似文献