We describe a convenient PCR-based protocol for in vitro recombination of homologous genes, thereby minimizing the rate of associated point mutations. High-fidelity recombination conditions were obtained using Vent DNA polymerase, which, in contrast to Taq DNA polymerase, shows significant proofreading activity and ranges among the slowest thermostable DNA polymerases, allowing tight control of...

Automating a software process both magnifies its strengths and accentuates its weaknesses. Automation can make an effective process more effective, but it can make a chaotic process even worse—and at considerable expense. Anyone who buys expensive tools to solve an ill-defined problem is likely to be disappointed. Unless procuring such tools is part of a thoughtful software process improvement ...

BACKGROUND Although mutations of several genes are associated with arrhythmogenic right ventricular cardiomyopathy (ARVC), the exact correlation between genotype and ventricular arrhythmia features remains unclear. This study was aimed to examine the possible association of the 9 known genes of ARVC with clinical and electrophysiological characteristics. METHODS AND RESULTS Ninety subjects di...

BACKGROUND A high prevalence of the R563Q mutation of the epithelial sodium channel β-subunit has been reported in South African hypertensives compared with unrelated normotensive controls. To delineate the effects of this mutation against a more uniform genetic background, this study investigated the association of the mutation with hypertension within affected kindreds. METHODS Forty-five i...

BACKGROUND A somatic mutation in the FOXL2 gene is reported to be present in almost all (97%; 86/89) morphologically defined, adult-type, granulosa-cell tumors (A-GCTs). This FOXL2 c.402C>G mutation changes a highly conserved cysteine residue to a tryptophan (p.C134W). It was also found in a minority of other ovarian malignant stromal tumors, but not in benign ovarian stromal tumors or unrelate...

Previous studies have demonstrated that some human endometrial carcinomas contain an activating point mutation in codon 12 of the Ki-ras protooncogene. To examine the hypothesis that this mutation may occur at an earlier stage of neoplastic progression in the endometrium, we analyzed 89 samples of premalignant endometrial hyperplasia and an additional 84 samples of endometrial carcinoma for poi...

Ten new and seventeen previously reported Enhanced S Cone Syndrome (ESCS) subjects were used to search for genetic heterogeneity. All subjects were diagnosed with ESCS on the basis of clinical, psychophysical and/or electroretinography testing using published criteria. Mutation analysis was performed on the NR2E3 nuclear receptor gene by single strand conformation analysis and direct sequencing...

BACKGROUND Clearance of hepatitis B virus (HBV) infection requires a good host immune response. Cytokines like tumor necrosis factor-alpha (TNF- alpha) may play a role in such immune response. Genetic changes in TNF-a gene promoter region are known to influence TNF- alpha expression. We therefore studied the role of one such mutation in chronic HBV infection. METHODS Presence of -308 G/A poly...

BACKGROUND Scientific data provide the evidence that secondary K-RAS mutations do not occur during anti-epidermal growth factor receptor therapy in colorectal cancer patients. This multicenter phase II prospective study aims to investigate the activity of a retreatment with a cetuximab-based therapy. PATIENTS AND METHODS We enrolled 39 irinotecan-refractory patients who had a clinical benefit...

The dinuclear cationic zirconium compound [{Zr( -C5H5)}2( -CH2)( -Cl)( -C5H4-C5H4)][BMe(C6F5)3] 1 reacts in dichloromethane at −78 °C with three equivalents of RNC (R= Bu, 2,6-Me2C6H3) via insertion into the Zr-methylene bond to give the new zirconium cationic species [{Zr( -C5H5)}2(Cl)(CNBu){ -[ -CN(Bu)-CH2-CN(Bu)}( -C5H4-C5H4)][BMe(C6F5)3] 2 and [{Zr( -C5H5)}2(Cl){CN(2,6-Me2C6H3)}[ -{CN(2,6-M...