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A method is described for performing sparse and stable gene selection from a number of unstable, but low cost, SVM-RFE units referred to as SVM-RFE subunits. Using a comprehensive simulation study, we show that the introduction of a consensus constraint with respect to variations in the policy of gene removal and a stability constraint with respect to perturbations in the training data can remarkably improve gene selection precision, dimensionality reduction ratio and stability of low cost SVM-RFE subunits still guaranteeing affordable computational costs. The method, which does not require the preselection of the number of selected genes, is divided into two stages. Multiple rough gene removal policies are first applied to multiple surrogate training datasets (spreading). Multiple consensus gene sets with respect to variations in the gene removal policy are then obtained and passed through a stability filter which selects the best performing gene set (despreading). Hence, while the consensus constraint performs strong dimensionality reduction at affordable computational costs, the stability constraint ensures acceptable indexes of gene selection stability and further dimensionality reduction. The method is validated on three benchmark microarray datasets. 相似文献
173.
《Planning》2016,(4)
目的 探讨1例糖原累积症Ia型(glycogen storage disease type Ia,GSD Ia)患者的临床特点及遗传机制。方法 详细收集患者临床资料,包括病史、体格检查、实验室检查结果。提取患者及其父母的外周血DNA,进行葡萄糖-6-磷酸酶催化亚基(glucose-6-phosphatase catalytic subunit,G6PC)基因5个外显子测序,对新突变进行蛋白功能预测。结果 患者为27岁男性,有低血糖、高乳酸血症、高尿酸血症和高脂血症的典型临床表现,肝穿刺活检支持GSD Ia。患者G6PC基因第2个外显子检测到c.248G>A(p.R83H)错义突变,第5个外显子检测到c.674T>C(p.L225P)错义突变,患者父亲和母亲分别是携带c.674T>C(p.L225P)及c.248G>A(p.R83H)突变的杂合子。采用Polyphen 2和SIFT软件对新发现的c.674T>C(p.L225P)突变蛋白功能进行预测,提示为致病突变,可能损害葡萄糖-6-磷酸酶蛋白功能。结论 G6PC基因的复合杂合突变是本例GSD Ia患者的致病基础,发现G6PC基因新的致病突变,拓宽了华人GSD Ia的致病基因谱。 相似文献
174.
Tae-Kyun Oh Sung Oh Seongdae Kim Jae Sung Park Nagarajan Vinod Kyung Min Jang Sei Chang Kim Chang Won Choi Suk-Min Ko Dong Kee Jeong Rajangam Udayakumar 《International journal of molecular sciences》2014,15(9):15571-15591
A full-length phytase gene (phy) of Aspergillus nidulans was amplified from the cDNA library by polymerase chain reaction (PCR), and it was introduced into a bacterial expression vector, pET-28a. The recombinant protein (rPhy-E, 56 kDa) was overexpressed in the insoluble fraction of Escherichia coli culture, purified by Ni-NTA resin under denaturing conditions and injected into rats as an immunogen. To express A. nidulans phytase in a plant, the full-length of phy was cloned into a plant expression binary vector, pPZP212. The resultant construct was tested for its transient expression by Agrobacterium-infiltration into Nicotiana benthamiana leaves. Compared with a control, the agro-infiltrated leaf tissues showed the presence of phy mRNA and its high expression level in N.
benthamiana. The recombinant phytase (rPhy-P, 62 kDa) was strongly reacted with the polyclonal antibody against the nonglycosylated rPhy-E. The rPhy-P showed glycosylation, two pH optima (pH 4.5 and pH 5.5), an optimum temperature at 45~55 °C, thermostability and broad substrate specificities. After deglycosylation by peptide-N-glycosidase F (PNGase-F), the rPhy-P significantly lost the phytase activity and retained 1/9 of the original activity after 10 min of incubation at 45 °C. Therefore, the deglycosylation caused a significant reduction in enzyme thermostability. In animal experiments, oral administration of the rPhy-P at 1500 U/kg body weight/day for seven days caused a significant reduction of phosphorus excretion by 16% in rat feces. Besides, the rPhy-P did not result in any toxicological changes and clinical signs. 相似文献
175.
AbstractBecause calcium phosphate (Ca–P) ceramics have been used as bone substitutes, it is necessary to investigate what effects the ceramics have on osteoblast maturation. We prepared three types of Ca–P ceramics with different Ca–P ratios, i.e. hydroxyapatite (HA), beta-tricalcium phosphate (β-TCP), and biphasic calcium phosphate (BCP) ceramics with dense-smooth and porous structures. Comprehensive gene expression microarray analysis of mouse osteoblast-like cells cultured on these ceramics revealed that porous Ca–P ceramics considerably affected the gene expression profiles, having a higher potential for osteoblast maturation. In the in vivo study that followed, porous Ca–P ceramics were implanted into rat skeletal muscle. Sixteen weeks after the implantation, more alkaline-phosphatase-positive cells were observed in the pores of hydroxyapatite and BCP, and the expression of the osteocalcin gene (an osteoblast-specific marker) in tissue grown in pores was also higher in hydroxyapatite and BCP than in β-TCP. In the pores of any Ca–P ceramics, 16 weeks after the implantation, we detected the expressions of marker genes of the early differentiation stage of chondrocytes and the complete differentiation stage of adipocytes, which originate from mesenchymal stem cells, as well as osteoblasts. These marker gene expressions were not observed in the muscle tissue surrounding the implanted Ca–P ceramics. These observations indicate that porous hydroxyapatite and BCP had a greater potential for promoting the differentiation of mesenchymal stem cells into osteoblasts than β-TCP. 相似文献
176.
A Systematic Structure–Activity Study of a New Type of Small Peptidic Transfection Vector Reveals the Importance of a Special Oxo-Anion-Binding Motif for Gene Delivery
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Dr. Sandra Junghänel Sarah Karczewski Sandra Bäcker Prof. Dr. Shirley K. Knauer Prof. Dr. Carsten Schmuck 《Chembiochem : a European journal of chemical biology》2017,18(22):2268-2279
We discovered a new class of artificial peptidic transfection vectors based on an artificial anion-binding motif, the guanidiniocarbonylpyrrole (GCP) cation. This new type of vector is surprisingly smaller than traditional systems, and our previous work suggested that the GCP group was important for promoting critical endosomal escape. We now present here a systematic comparison of similar DNA ligands featuring our GCP oxo-anion-binding motif with DNA ligands only consisting of naturally occurring amino acids. Structure–activity studies showed that the artificial binding motif clearly outperformed natural amino acids such as histidine, lysine, and arginine. It improved the ability to shuttle foreign genetic material into cells, yet successfully mediated endosomal escape. Also, plasmids that were complexed by our artificial ligands were stabilized against cytosolic degradation to some extent. This resulted in the successful expression of plasmid information (comparable to gold standards such as polyethyleneimine). Hence, our study clearly demonstrates the importance of the tailor-made GCP anion-binding site for efficient gene transfection. 相似文献
177.
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179.
Julia Welzenbach Christiane Neuhoff Hanna Heidt Mehmet Ulas Cinar Christian Looft Karl Schellander Ernst Tholen Christine Gro?e-Brinkhaus 《International journal of molecular sciences》2016,17(9)
The aim of this study was to integrate multi omics data to characterize underlying functional pathways and candidate genes for drip loss in pigs. The consideration of different omics levels allows elucidating the black box of phenotype expression. Metabolite and protein profiling was applied in Musculus longissimus dorsi samples of 97 Duroc × Pietrain pigs. In total, 126 and 35 annotated metabolites and proteins were quantified, respectively. In addition, all animals were genotyped with the porcine 60 k Illumina beadchip. An enrichment analysis resulted in 10 pathways, amongst others, sphingolipid metabolism and glycolysis/gluconeogenesis, with significant influence on drip loss. Drip loss and 22 metabolic components were analyzed as intermediate phenotypes within a genome-wide association study (GWAS). We detected significantly associated genetic markers and candidate genes for drip loss and for most of the metabolic components. On chromosome 18, a region with promising candidate genes was identified based on SNPs associated with drip loss, the protein “phosphoglycerate mutase 2” and the metabolite glycine. We hypothesize that association studies based on intermediate phenotypes are able to provide comprehensive insights in the genetic variation of genes directly involved in the metabolism of performance traits. In this way, the analyses contribute to identify reliable candidate genes. 相似文献
180.