Experimental diagnostic of sequence-variant dynamic perturbations revealed by broadband dielectric spectroscopy

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Restriction fragment length variants in the marsupialSminthopsis crassicaudata

A fully pedigreed colony of the dasyurid marsupialSminthopsis crassicaudata has provided material for establishing two panels of DNA samples: a broad-based test panel and a two-generation family panel. These have been used to search for genetic markers in the form of restriction fragment length variants. The molecular probes—pSG-2H, a region of theS. crassicaudata embryonic -globin gene; pB8.BS, a region of the human ubiquitin gene, and p3-21a1:1, a region of the processed pseudogene of phosphoglycerate kinase-1 of the macropodid marsupialMacropus robustus—were hybridized to Southern blots ofEcoR1-digested DNA from the panels. Analysis of these blots when probed with pSG-2H provided evidence of two alleles segregating at a singleEcoR1 site. Analysis of the same blots when probed with pB8.BS suggested allelic variation at two closely linkedEcoR1 sites. Probing the blots with p3-21a1:1 produced a complex pattern of bands resembling DNA fingerprints. The presence of a 12.3-kb band was found to conform to a simple autosomal dominant mode of inheritance. Analysis of the family data, for each probe, revealed no significant departure from Mendelian inheritance. This work has provided additional genetic markers that will enhance the use ofS. crassicaudata as a model marsupial species and has demonstrated that a high level of genetic variability has been maintained in the marsupial colony.This project was supported by grants from the Australian Research Council and the University of Adelaide.     

Alternate Junctions and Microheterogeneity of Tlr1, a Developmentally Regulated DNA Rearrangement in Tetrahymena thermophila

ABSTRACT. A large number of developmentally regulated DNA rearrangements occur during the development of the macronucleus in Tetrahymena thermophila , Tlr1 is a deletion element which has large inverted repeats near the rearrangement junctions and deletes more than 13 kbp of internal DNA. Previous analysis of caryonidal lines revealed alternate left junctions for the Tlr1 rearrangement in B strain cells. We show here that C2 strain Tetrahymena also use alternate rearrangement junctions. We have mapped and sequenced two additional rearrangement variants and find that both the left and right can vary over a range of approximately 200 bp. We also demonstrate the presence of sequence microheterogeneity in the most commonly found Tlr1 rearrangement product.     

RBPMS不同剪接体的真核表达和亚细胞定位

目的：构建具有多种剪接形式的RNA结合蛋白（RBPMS）基因的真核表达载体,并在真核细胞中表达,确定不同形式的RBPMS在细胞中的定位。方法：采用PCR技术从人卵巢cDNA文库中扩增RBPMS基因的几种完整编码序列（命名为RBPl～RBP4）,克隆到带绿色荧光蛋白标签的pEGFP-C1表达载体上,转染人胚肾细胞293T,Western印迹鉴定RBPMS的表达,并利用激光共聚焦显微镜观察RBPMS不同剪接体在细胞中的定位。结果：限制性内切酶分析和DNA序列测定表明构建的重组表达载体正确,Western印迹实验证明RBP1～RBP4表达成功。通过激光共聚焦显微镜观察,RBP1／4围绕胞核在核膜的周围呈聚集状分布;RBP2则在细胞质和细胞核中均有分布,但会出现斑点状聚集;RBP3呈半月状紧密分布在细胞核周围;RBPMS中的RNA识别基序缺失后,这种现象消失,与空载体对照类似,在细胞核和细胞质中均有分布。结论：构建并表达了RBPMS基因的真核表达载体,RBPMS不同剪接体及RNA识别基序缺失后具有不同的亚细胞分布模式,提示具有不同的功能。     

Glucose-6-phosphate dehydrogenase (G6PD) Guantánamo and G6PD Caujerí: Two new glucose-6-phosphate dehydrogenase-deficient variants found in Cuba

Glucose-6-phosphate dehydrogenase (G6PD) deficiency was identified in two children who were studied because of hemolytic episodes. The electrophoretic and kinetic properties of the mutant enzymes allowed us to conclude that both of them were new variants. They were named G6PD Guantánamo and G6PD Caujerí.     

Multi-omics approach dissects cis-regulatory mechanisms underlying North Carolina macular dystrophy,a retinal enhanceropathy

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Evaluation of T cell responses to naturally processed variant SARS-CoV-2 spike antigens in individuals following infection or vaccination

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Chromogranin A in a Cohort of Pheochromocytomas and Paragangliomas: Usefulness at Diagnosis and as an Early Biomarker of Recurrence

ObjectiveTo evaluate the usefulness of chromogranin A (CgA) in the management of patients with pheochromocytomas (PHEOs) and paragangliomas (PGLs).MethodsWe retrospectively reviewed the charts of 132 patients with confirmed PHEOs/PGLs (PPGLs) followed at our medical center. CgA was measured in 80 patients at diagnosis. The exclusion criteria removed 19 of these patients. Five patients with relapses were also analyzed.ResultsOur cohort of 61 patients included 34 PHEOs, 14 head and neck PGLs, and 13 thoracoabdominal PGLs. CgA levels were elevated in 53 of 61 patients (86.9%) at diagnosis: 33 of 34 (97.1%) PHEOs, 9 of 14 (64.3%) head and neck paragangliomas, and 11 of 13 (84.6%) thoracoabdominal paragangliomas. For 8 of 13 (61.5%) nonfunctional PPGLs (5 head and neck paragangliomas and 3 thoracoabdominal paragangliomas), increased CgA levels showed potential as a tumor marker during follow-up. Of 10 patients with malignant PPGLs, only 1 had normal CgA levels (10.0%). Among 54 patients with PPGLs who underwent genetic testing, elevated CgA levels were positive in 73.7% of patients carrying a germline genetic variant (pathogenic and of unknown significance) versus 91.4% of patients without a known germline variant. We also report 5 PPGL cases with increased CgA levels as the first detectable marker of tumoral recurrence or progression preceding other biochemical markers or imaging.ConclusionCgA is a sensitive marker for the diagnosis of PHEO (97.1%) and thoracoabdominal paraganglioma (84.6%). CgA may be useful in the follow-up of nonfunctional PGLs and may also play a complementary role in the early detection of recurrence in secreting PPGLs.     

Genomic monitoring of SARS-CoV-2 uncovers an Nsp1 deletion variant that modulates type I interferon response

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