Rapid detection of epidermal growth factor receptor mutations with multiplex PCR and primer extension in lung cancer

Epidermal growth factor receptor (EGFR) kinase domain mutations hyperactivate the kinase and confer kinase addiction of the non-small-cell lung cancer (NSCLC) tumor cells. Almost all of these mutations are located within exons 18-21. The -216 single nucleotide polymorphism in the promoter region is associated with increased EGFR production. We present a method for detecting these common mutations in 81 cases of NSCLC. The protocol is based on the multiplex amplification of promoter region and exons 18-21 of the EGFR genes in a single tube, followed by primer extension of the PCR products using various sizes of primers to detect base changes at -216 promoter region and codons 719, 746-750, 790, 858 of the EGFR gene. We compared the results with that from direct sequencing for detecting EGFR mutations in 81 cases of NSCLC. The two methods identified the same 26 mutations, but our method is superior to direct sequencing in terms of the amount of work and time required. We presented a simple and fast method to detect mutations of EGFR genes in NSCLC

Benefits of exercise training and the correlation between aerobic capacity and functional outcomes and quality of life in elderly patients with coronary artery disease

AbstractCardiopulmonary exercise training is beneficial to people with coronary artery disease (CAD). Nevertheless, the correlation between aerobic capacity, and functional mobility and quality of life in elderly CAD patients is less addressed. The purpose of the current study is to investigate the beneficial effects of exercise training in elderly people with CAD, integrating exercise stress testing, functional mobility, handgrip strength, and health-related quality of life. Elderly people with CAD were enrolled from the outpatient clinic of a cardiac rehabilitation unit in a medical center. Participants were assigned to the exercise training group (N = 21) or the usual care group (N = 15). A total of 36 sessions of exercise training, completed in 12 weeks, was prescribed. Echocardiography, exercise stress testing, the 6-minute walking test, Timed Up and Go test, and handgrip strength testing were performed, and the Short-Form 36 questionnaire (SF-36) was administered at baseline and at 12-week follow-up. Peak oxygen consumption improved significantly after training. The heart rate recovery improved from 13.90/minute to 16.62/minute after exercise training. Functional mobility and handgrip strength also improved after training. Significant improvements were found in SF-36 physical function, social function, role limitation due to emotional problems, and mental health domains. A significant correlation between dynamic cardiopulmonary exercise testing parameters, the 6-minute walking test, Timed Up and Go test, handgrip strength, and SF-36 physical function and general health domains was also detected. Twelve-week, 36-session exercise training, including moderate-intensity cardiopulmonary exercise training, strengthening exercise, and balance training, is beneficial to elderly patients with CAD, and cardiopulmonary exercise testing parameters correlate well with balance and quality of life

Clinical Study of Uric Acid Urolithiasis

Uric acid urolithiasis develops from various causes. To investigate the clinical and biochemical presentation of patients with uric acid urolithiasis, a retrospective study was designed. A total of 46 cases were enrolled between January 2004 and December 2005. The compositions of the stones were analyzed by infrared spectrophotometry. There were 39 males (84.8%) and seven females (15.2%), with a mean age of 61.5 ± 10.6 years and mean body mass index (BMI) of 26.7 ± 3.1 kg/m2. The stone location was kidney in 10 (21.7%), ureter in 22 (41.8%), and bladder in 14 (30.5%). Multiple stones were diagnosed in 36 patients (78.3%). Pre-existing comorbidities included diabetes mellitus in 11 patients (23.9%), hypertension in 23 (50%), gout in 13 (28.2%), and benign prostatic hyperplasia in 14 (30.4%). Mean serum creatinine and uric acid was 1.6 ± 0.6 mg/dL and 7.6 ± 1.8 mg/dL, respectively. There were 27 patients (58%) with creatinine > 1.4 mg/dL. The mean urinary pH was 5.42 ± 0.46. Patients with uric acid urolithiasis were predominantly male, older, with higher BMI, multiple stone presentation, with lower urinary pH, and hyperuricemia. Exacerbation of the renal function should also be of concern because of the high proportion of patients with renal insufficiency diagnosed in this study

Do beta-adrenergic blocking agents increase asthma exacerbation? A network meta-analysis of randomized controlled trials

Beta-adrenergic blocking agents (abbreviated as beta-blockers) have been used for treating various cardiovascular diseases. However, the potential for asthma exacerbation is one of the major adverse effects of beta-blockers. This study aimed to compare the level of risk for an asthma attack in patients receiving various beta-blockers. We searched for randomized controlled trials (RCTs) of either placebo-controlled or active-controlled design. The current network meta-analysis (NMA) was conducted under a frequentist model. The primary outcome was the incidence of asthmatic attack. A total of 24 RCTs were included. Overall NMA revealed that only oral timolol [risk ratio (RR) = 3.35 (95% confidence interval (CI) 1.04–10.85)] and infusion of propranolol [RR = 10.19 (95% CI 1.29–80.41)] were associated with significantly higher incidences of asthma attack than the placebo, whereas oral celiprolol [RR = 0.39 (95% CI 0.04–4.11)], oral celiprolol and propranolol [RR = 0.46 (95% CI 0.02–11.65)], oral bisoprolol [RR = 0.46 (95% CI 0.02–11.65)], oral atenolol [RR = 0.51 (95% CI 0.20–1.28)], infusion of practolol [RR = 0.80 (95% CI 0.03–25.14)], and infusion of sotalol [RR = 0.91 (95% CI 0.08–10.65)] were associated with relatively lower incidences of asthma attack than the placebo. In participants with a baseline asthma history, in addition to oral timolol and infusion of propranolol, oral labetalol, oxprenolol, propranolol, and metoprolol exhibited significantly higher incidences of asthma attack than did the placebo. In conclusion, oral timolol and infusion of propranolol were associated with a significantly higher risk of developing an asthma attack in patients, especially in those with a baseline asthma history, and should be avoided in patients who present a risk of asthma

Transcriptomic analyses of regenerating adult feathers in chicken

Transcriptome Expression Data. Table of mapped reads to Galgal4 transcripts for all 15 data sets. FPKM (Fragments per kilobase of exon per million fragments mapped): normalized transcript abundance values for each gene in the indicated tissues. (CSV 1314 kb

Roles of cysteines Cys115 and Cys201 in the assembly and thermostability of grouper betanodavirus particles

The virus-like particle (VLP) assembled from capsid subunits of the dragon grouper nervous necrosis virus (DGNNV) is very similar to its native T = 3 virion. In order to investigate the effects of four cysteine residues in the capsid polypeptide on the assembly/dissociation pathways of DGNNV virions, we recombinantly cloned mutant VLPs by mutating each cysteine to destroy the specific disulfide linkage as compared with thiol reduction to destroy all S–S bonds. The mutant VLPs of C187A and C331A mutations were similar to wild-type VLPs (WT-VLPs); hence, the effects of Cys187 and Cys331 on the particle formation and thermostability were presumably negligible. Electron microscopy showed that either C115A or C201A mutation disrupted de novo VLP formation significantly. As shown in micrographs and thermal decay curves, β-mercaptoethanol-treated WT-VLPs remained intact, merely resulting in lower tolerance to thermal disruption than native WT-VLPs. This thiol reduction broke disulfide linkages inside the pre-fabricated VLPs, but it did not disrupt the appearance of icosahedrons. Small dissociated capsomers from EGTA-treated VLPs were able to reassemble back to icosahedrons in the presence of calcium ions, but additional treatment with β-mercaptoethanol during EGTA dissociation resulted in inability of the capsomers to reassemble into the icosahedral form. These results indicated that Cys115 and Cys201 were essential for capsid formation of DGNNV icosahedron structure in de novo assembly and reassembly pathways, as well as for the thermal stability of pre-fabricated particles

Métodos multivariados aplicados ao melhoramento genético do feijoeiro visando ao aumento da tolerância ao estresse osmótico e biofortificação de grãos

O feijoeiro (Phaseolus vulgaris L.) é uma cultura agrícola muito importante economicamente e nutricionalmente para a população brasileira e necessita de metodologias simples e eficazes que auxiliem o processo de melhoramento genético. As técnicas empregadas devem minimizar os efeitos indesejáveis da multicolinearidade entre as características estudadas durante o processo de seleção. A produção de sementes de feijão, normalmente, é limitada pela escassez hídrica e solos salinos. No entanto, devido a grande variabilidade genética, característica da espécie, é possível encontrar materiais genéticos mais tolerantes a esses estresses osmóticos. A germinação e o desenvolvimento inicial da plântula são fases críticas e desta maneira é importante selecionar os matérias genéticos mais tolerantes nestas fases. Além de selecionar genótipos tolerantes é necessário selecionar materiais genéticos que sejam ricos nutricionalmente, principalmente, em relação à composição mineralógica. Os principais objetivos almejados com este trabalho foram reduzir a multicolinearidade e selecionar genótipos para a tolerância ao estresse osmótico e a biofortificação dos grãos do feijoeiro, com base nos valores genéticos. Desta maneira, foram utilizadas duas técnicas para reduzir a influência da multicolinearidade: o descarte de variáveis redundantes pelas variáveis canônicas, e o uso das análises de fatores para reduzir o número de variáveis. As variáveis analisadas foram: porcentagem de germinação e de plântulas normais, tempo médio de germinação, índice de velocidade de germinação, comprimentos de raiz e de hipocótilo, massas seca de raiz e da parte aérea, razão raiz/parte aérea e o produto da porcentagem de plântulas normais pelo comprimento das plântulas. Avaliou-se também a composição mineralógica dos grãos em relação à concentração de cálcio, ferro, zinco, potássio, magnésio, manganês e fósforo. Adicionalmente, para estimar os parâmetros e os valores genéticos realizou-se análise via modelos mistos, utilizando-se a técnica de REML/BLUP. Os genótipos foram selecionados com base da média genética, estabilidade e adaptabilidade, utilizando-se a técnica da média harmônica da performance relativa dos valores genéticos. Os genótipos que apresentaram as maiores tolerâncias, adaptabilidade e estabilidade quanto aos estresses osmóticos foram: CNFC 15466, CNFC 15462, CNFC 15630, BRS Valente, Capixaba Precoce, CNFP 15290, CNFP 15292 e CNFP 15302. Enquanto os genótipos mais ricos e divergentes geneticamente do grupo comercial carioca foram: CNFC 15475 e CNFC 15625, e do grupo comercial preto foram: CNFP 15310 e CNFP 15304. Conclui-se que a utilização de técnicas multivariadas facilita a seleção de genótipos promissores como parentais na formação de linhagens tolerantes ao estresse osmótico e biofortificados. Palavras-chave: feijoeiro comum; seleção de genótipos; estresse hídrico e salino; multicolinearidade; composição mineral

Performance of CMS muon reconstruction in pp collision events at sqrt(s) = 7 TeV

The performance of muon reconstruction, identification, and triggering in CMS has been studied using 40 inverse picobarns of data collected in pp collisions at sqrt(s) = 7 TeV at the LHC in 2010. A few benchmark sets of selection criteria covering a wide range of physics analysis needs have been examined. For all considered selections, the efficiency to reconstruct and identify a muon with a transverse momentum pT larger than a few GeV is above 95% over the whole region of pseudorapidity covered by the CMS muon system, abs(eta) < 2.4, while the probability to misidentify a hadron as a muon is well below 1%. The efficiency to trigger on single muons with pT above a few GeV is higher than 90% over the full eta range, and typically substantially better. The overall momentum scale is measured to a precision of 0.2% with muons from Z decays. The transverse momentum resolution varies from 1% to 6% depending on pseudorapidity for muons with pT below 100 GeV and, using cosmic rays, it is shown to be better than 10% in the central region up to pT = 1 TeV. Observed distributions of all quantities are well reproduced by the Monte Carlo simulation.Comment: Replaced with published version. Added journal reference and DO

Performance of CMS muon reconstruction in pp collision events at sqrt(s) = 7 TeV

The performance of muon reconstruction, identification, and triggering in CMS has been studied using 40 inverse picobarns of data collected in pp collisions at sqrt(s) = 7 TeV at the LHC in 2010. A few benchmark sets of selection criteria covering a wide range of physics analysis needs have been examined. For all considered selections, the efficiency to reconstruct and identify a muon with a transverse momentum pT larger than a few GeV is above 95% over the whole region of pseudorapidity covered by the CMS muon system, abs(eta) < 2.4, while the probability to misidentify a hadron as a muon is well below 1%. The efficiency to trigger on single muons with pT above a few GeV is higher than 90% over the full eta range, and typically substantially better. The overall momentum scale is measured to a precision of 0.2% with muons from Z decays. The transverse momentum resolution varies from 1% to 6% depending on pseudorapidity for muons with pT below 100 GeV and, using cosmic rays, it is shown to be better than 10% in the central region up to pT = 1 TeV. Observed distributions of all quantities are well reproduced by the Monte Carlo simulation.Comment: Replaced with published version. Added journal reference and DO

X-ray emission from the Sombrero galaxy: discrete sources

We present a study of discrete X-ray sources in and around the bulge-dominated, massive Sa galaxy, Sombrero (M104), based on new and archival Chandra observations with a total exposure of ~200 ks. With a detection limit of L_X = 1E37 erg/s and a field of view covering a galactocentric radius of ~30 kpc (11.5 arcminute), 383 sources are detected. Cross-correlation with Spitler et al.'s catalogue of Sombrero globular clusters (GCs) identified from HST/ACS observations reveals 41 X-rays sources in GCs, presumably low-mass X-ray binaries (LMXBs). We quantify the differential luminosity functions (LFs) for both the detected GC and field LMXBs, whose power-low indices (~1.1 for the GC-LF and ~1.6 for field-LF) are consistent with previous studies for elliptical galaxies. With precise sky positions of the GCs without a detected X-ray source, we further quantify, through a fluctuation analysis, the GC LF at fainter luminosities down to 1E35 erg/s. The derived index rules out a faint-end slope flatter than 1.1 at a 2 sigma significance, contrary to recent findings in several elliptical galaxies and the bulge of M31. On the other hand, the 2-6 keV unresolved emission places a tight constraint on the field LF, implying a flattened index of ~1.0 below 1E37 erg/s. We also detect 101 sources in the halo of Sombrero. The presence of these sources cannot be interpreted as galactic LMXBs whose spatial distribution empirically follows the starlight. Their number is also higher than the expected number of cosmic AGNs (52+/-11 [1 sigma]) whose surface density is constrained by deep X-ray surveys. We suggest that either the cosmic X-ray background is unusually high in the direction of Sombrero, or a distinct population of X-ray sources is present in the halo of Sombrero.Comment: 11 figures, 5 tables, ApJ in pres