Functional correction of short-chain acyl-CoA dehydrogenase deficiency in transgenic mice: implications for gene therapy of human mitochondrial enzyme deficiencies

We report the therapeutic effects of liver-specific expression of a short-chain acyl-CoA dehydrogenase (SCAD) transgene in the SCAD- deficient mouse model. Transgenic mice were produced with a rat albumin promoter/enhancer driving a mouse SCAD minigene (ALB-SCAD) on both the SCAD normal genetic background and a SCAD-deficient background. In three transgenic lines produced on the SCAD-deficient background, recombinant SCAD activity and antigen in liver mitochondria were found up to 7-fold of normal control values. All three lines showed a markedly reduced organic aciduria and fatty liver, which are sensitive indicators of the metabolic abnormality seen in this disease found in children. We found no detrimental effects of high liver SCAD expression in transgenic mice on either background. These studies provide important basic and practical therapeutic information for the potential gene therapy of nuclear-encoded mitochondrial enzyme deficiencies, as well as insights into the mechanisms of the disease.      

丙氧鸟苷对转TK基因的人外周血单个核细胞的毒性作用

目的 探讨减少异体干细胞移植中移植抗宿主病（GVHD）,同时最大限度提高移植抗白血病（GVL）效应的有效方法。方法 采用逆转录病毒介导的基因转移方法将I型单纯疱疹病毒胸苷激酶（HSV－TK）基因、绿色荧光蛋白（GFP）基因及抗新霉素（NeoR）基因插入人外周血单个核细胞（PBMC）,MTT法测定丙氧鸟苷（GCV）对转化细胞抑制率。结果 转化细胞表达GFP,且主要为T淋巴细胞,占11．4％,而且被转化的T淋巴细胞中,CD4阳性细胞转化率较高,占7．6％,CD8,CD19,CD33阳性细胞转化率分别为2．9％,2．1％和4．7％,PCR鉴定表明,转染的人外周血单个核细胞DNA中整合中有NeoR基因,MTT法测定丙氧鸟苷（GCV）对转化细胞与未转化细胞抑制率,显示转化细胞生长明显受抑。结论 异体干细胞移植后,如产生严重GVHD应用GCV选择性清除HSV－TK基因转导的T淋巴细胞,使控制GVHD已成可能。     

Extensive CFTR sequencing through NGS in Brazilian individuals with cystic fibrosis: unravelling regional discrepancies in the country

BackgroundThe Brazilian population has a tri-hybrid composition with a high degree of ethnic admixture. We hypothesized that Brazilian individuals with CF from different Brazilian regions have a specific distribution of CFTR variants.MethodsIndividuals with CF with data available in the Patient Registry and without an established genotype were submitted to CFTR sequencing by Next Generation Sequencing (NGS) methodology, and results were anonymously incorporated to the Registry Database. Genotyping results were expressed as ‘positive’, ‘inconclusive’ or ‘negative’. Logistic regression models were performed to investigate the association between demographic/clinical variables and genotyping results. Mediation analysis was conducted to estimate direct and indirect effects of Brazilian region on a binary positive genotyping response.ResultsIn October 2017, data from 4,654 individuals with CF were available, and 3,104(66.7%) of them had a genotyping result. A total of 236 variants (114 new variants) were identified, with F508del identified in 46% of the alleles tested. Genotyping revealed 2,002(64.5%) individuals positive, 757(24.4%) inconclusive and 345(11.1%) negative. Distribution of genotype categories was markedly different across Brazilian Regions, with greater proportions of negative individuals in the North (45%) and Northeast (26%) regions. Newborn screening (CF-NBS) and age at diagnosis were identified as mediators of the effect of Brazilian region on a positive genotyping result.ConclusionsThis large initiative of CFTR genotyping showed significant regional discrepancies in Brazil, probably related to socio-economic conditions, lack of adequate CF-NBS and poor access to reliable sweat testing. These results may be useful to indicate Regions where CF care demands more attention.     

Irritable bowel syndrome: A technical review for practice guideline development

This article has no abstract. To view the article, select the "View Print Version (PDF)" link above.     

Epidemiology, values, and the meaning of paradigm

Use of the term paradigm has recently reached the epidemiological milieu. This article provides a critical analysis of a classification of "epidemiological paradigms" as proposed by Almeida-Filho. It was initially ascertained that values were not considered fundamental elements for the construction of paradigms, and that they therefore distanced themselves from the Kuhnian matrix. Systematization sought to unite antithetical tendencies and thus seemed closer to the Foucaltian epistéme. Finally, hegemony was considered a more appropriate term than paradigm for systematizing epidemiological periods, since the values of the different watersheds are committed to particularity (principally social class).     

左旋卡尼汀治疗不稳定型心绞痛疗效观察

目的:探讨左旋卡尼汀治疗不稳定性心绞痛的临床疗效.方法:86例不稳定性心绞痛患者随机分为两组,每组43例.在常规治疗基础上,治疗组应用左旋卡尼汀,对照组应用极化液.结果:心绞痛缓解有效率治疗组为93.02%,对照组心绞痛缓解有效率为65.12%(P＜0.01);静息心电图缺血性ST段恢复有效率治疗组为76.74%,对照组为48.84%(P＜0.01).结论:左旋卡尼汀辅助治疗不稳定性心绞痛的是一种安全有效的方法,它能改善缺血心肌的能量代谢.     

Monocyte-dependent regulation of T lymphocyte activation through CD98

CD98 is a 125 kDa heterodimer, which is strongly expressed on the surface of activated and proliferating cells. Its expression is strikingly regulated during T cell differentiation and activation, but the role of CD98 during T lymphocyte responses is not yet understood. We report here that proliferation of resting peripheral blood mononuclear cells (PBMC) induced by lectin, superantigen (SAg) or conventional antigens was blocked by anti-CD98 heavy chain (CD98hc) mAb. In contrast, anti-CD98hc did not block responses of T cell clones or lines. Anti-CD98hc inhibited IL-2 receptor expression and progression of T cells from G1 to S phase, but did not reduce expression of the IL-2 gene. Anti-CD98hc mAb did not regulate the initial activation events involving the TCR and co-receptor structures, but instead inhibited T lymphocyte responses even when added 18 h or more after the activation stimulus. Further experiments demonstrated that anti-CD98 was not directly affecting T cells in this system, but was instead acting on accessory cells. This was supported using a novel xenogeneic system that takes advantage of the lack of xenoreactivity of purified human T cells against mouse splenocytes. Despite absence of a direct xenoresponse to murine spleen cells, human T cells were activated by SAg presented by murine splenic antigen-presenting cells (APC). Murine anti-human CD98hc did not block T cell proliferation in this system. Furthermore, responses using monocyte-depleted PBMC as APC were not blocked by anti-CD98hc. Taken together, the present data suggests that triggering of human monocyte CD98 can suppress T cell proliferation by a process that halts progression through the cell cycle of recently activated T lymphocytes. This may represent a novel pathway for monocyte regulation of T cell activation.