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Implementation and Validation of the Roche Light Cycler 480 96-Well Plate Platform as a Real-Time PCR Assay for the Quantitative Detection of Cytomegalovirus (CMV) in Clinical Specimens Using the Luminex MultiCode ASRs System.
Allogenic stem-cell therapies benefit patients in the treatment of multiple diseases; however, the side effects of stem-cell therapies (SCT) derived from the concomitant use of immune suppression agents often include triggering infection diseases. Thus, analysis is required to improve the detection of pathogen infections in SCT. We develop a polymerase chain reaction (PCR)-based methodology for the qualitative real-time DNA detection of cytomegalovirus (CMV), with reference to herpes simplex virus types 1 (HSVI), Epstein-Barr virus (EBV), and varicella-zoster virus (VZV) in blood, urine, solid tissues, and cerebrospinal fluid. This real-time PCR of 96-well plate format provides a rapid framework as required by the Food and Drug Administration (FDA) for clinical settings, including the processing of specimens, reagent handling, special safety precautions, quality control criteria and analytical accuracy, precisely reportable range (analyst measurement range), reference range, limit of detection (LOD), analytical specificity established by interference study, and analyte stability. Specifically, we determined the reportable range (analyst measurement range) with the following criteria: CMV copies ≥200 copies/mL; report copy/mL value; CMV copies ≤199 copies/mL; report detected but below quantitative range; CMV copies = 0 with report <200 copies/mL. That is, with reference range, copy numbers (CN) per milliliter (mL) of the LOD were determined by standard curves that correlated Ct value and calibrated standard DNA panels. The three repeats determined that the measuring range was 1E2~1E6 copies/mL. The standard curves show the slopes were within the range -2.99 to -3.65 with R2 ≥ 0.98. High copy (HC) controls were within 0.17-0.18 log differences of DNA copy numbers; (2) low copy (LC) controls were within 0.17-0.18 log differences; (3) LOD was within 0.14-0.15 log differences. As such, we set up a fast, simple, inexpensive, sensitive, and reliable molecular approach for the qualitative detection of CMV pathogens. Conclusion: This real-time PCR of the 96-well plate format provides a rapid framework as required by the FDA for clinical settings
Catabolite repression of the citST two-component system in Bacillus subtilis
In Bacillus subtilis, expression of the citrate transporter CitM is under strict control. Transcription of the citM gene is induced by citrate in the medium mediated by the CitS-CitT two-component system and repressed by rapidly degraded carbon sources mediated by carbon catabolite repression (CCR). In this study, we demonstrate that citST genes are part of a bicistronic operon. The promoter region was localized in a stretch of 58 base pairs upstream of the citS gene by deletion experiments. Transcription of the operon was repressed in the presence of glucose by the general transcription factor CcpA. A distal consensus cre site in the citS-coding sequence was implicated in the mechanism of repression. Furthermore, this repression was relieved in Bacillus subtilis mutants deficient in CcpA or Hpr/Crh, components essential to CCR. Thus, we demonstrate that CCR represses the expression of the citST operon, which is responsible for the induction of citM, through the cre site located 1326 bp from transcriptional start site of citST
The link between linguistic subordination and linguistic inferiority complexes: English as a second language migrants in Australia
Aims and objectives/purpose/research questions: This article aims to explore the link between linguistic subordination and linguistic inferiority complexes in the context of English as a second language (ESL) migrants in Australia. We address the following research questions: (a) To what extent and how do ESL migrants in Australia suffer from linguistic subordination? (b) To what extent and how is this linguistic subordination linked to linguistic inferiority complexes for ESL-migrants in Australia? (c) What are the main social implications of the link between linguistic subordination and linguistic inferiority complexes for ESL migrants? Design/methodology/approach: The study involved the participation of 150 participants who were observed using linguistic ethnography. The participants engaged in interviews and focus group discussions so that we could explore the psychological consequences of linguistic subordination that they encountered. Data and analysis: The interviews and focus group discussions were transcribed and analysed with consideration to the context, as well as the participants’ utterances and paralanguage. The study followed alternative quality criteria. Findings/conclusions: Many ESL migrants in Australia face linguistic subordination for the way they speak English. Consequently, these migrants suffer from linguistic inferiority complexes – psychological and emotional damages, which result in self-marginalisation, self-vindication, loss of sense of belonging, social withdrawal, fear, anxiety and the erosion of self-confidence. We find that there is a direct link between linguistic subordination and linguistic inferiority complexes. Originality: This article addresses a gap that exists regarding the link between linguistic subordination and the development of linguistic inferiority complexes, discusses how this has real-life consequences for ESL migrants, and explores how this may be overcome. Significance/implications: ESL migrants need support so that they can cope with their experiences of linguistic subordination, thus helping them better deal with issues related to linguistic inferiority complexes. In addition, the broader society needs greater education on how their comments and actions can affect the wellbeing of others
Do About Half the Top Quarks at FNAL Come From Gluino Decays?
We argue that it is possible to make a consistent picture of FNAL data
including the production and decay of gluinos and squarks. The additional cross
section is several pb, about the size of that for Standard Model (SM) top quark
pair production. If the stop squark mass is small enough, about half of the top
quarks decay to stop squarks, and the loss of SM top quark pair production rate
is compensated by the supersymmetric processes. This behavior is consistent
with the reported top quark decay rates in various modes and other aspects of
the data, and suggests several other possible decay signatures. This picture
can be tested easily with more data, perhaps even with the data in hand, and
demonstrates the potential power of a hadron collider to determine
supersymmetric parameters. It also has implications for the top mass
measurement and the interpretation of the LEP excess.Comment: 18 pages, including 4 Postscript figures, uses epsf.tex, also
available at http://www.hep.anl.gov/theory/mrenna
Socioeconomic Impacts on Survival Differ by Race/Ethnicity among Adolescents and Young Adults with Non-Hodgkin's Lymphoma
Shorter survival has been associated with low socioeconomic status (SES) among elderly non-Hodgkin's lymphoma (NHL) patients; however it remains unknown whether the same relationship holds for younger patients. We explored the California Cancer Registry (CCR), to investigate this relationship in adolescent and young adult (AYA) NHL patients diagnosed from 1996 to 2005. A case-only survival analysis was conducted to examine demographic and clinical variables hypothesized to be related to survival. Included in the final analysis were 3,489 incident NHL cases. In the multivariate analyses, all-cause mortality (ACM) was higher in individuals who had later stage at diagnosis (P < .05) or did not receive first-course chemotherapy (P < .05). There was also a significant gradient decrease in survival, with higher ACM at each decreasing quintile of SES (P < .001). Overall results were similar for lymphoma-specific mortality. In the race/ethnicity stratified analyses, only non-Hispanic Whites (NHWs) had a significant SES-ACM trend (P < .001). Reduced overall and lymphoma-specific survival was associated with lower SES in AYAs with NHL, although a significant trend was only observed for NHWs
Enfermedad funcional tiroidea en la poblaciĂłn de edad avanzada
ObjetivoDescribir la prevalencia de enfermedad funcional tiroidea (EFT) y otras afecciones asociadasa ella en una poblaciĂłn de edad avanzadaDiseñoEstudio descriptivo transversalEmplazamientoCentro de atenciĂłn primaria urbanoPacientesMuestra representativa de toda la poblacion atendida con edad . 60 anosMediciones principalesVariables demográficas, antecedentes clĂnicos de enfermedad tiroidea y afecciones asociadas a ella, Ăndice de masa corporal, tests reducidos para el diagnĂłstico de depresiĂłn y ansiedad, Mini-Mental Test, electrocardiograma, determinaciĂłn de colesterol total y colesterol unido a lipoproteĂnas de baja densidad (cLDL) y determinaciĂłn de tirotropina y tiroxina libre si Ă©sta estaba alteradaResultadosSe estudio a 192 personas, un 56% mujeres, con un 53% de 60–69 anos y un 12% con edad superior a 79 anos. Un 10% tenia antecedentes de EFT previa. La prevalencia de EFT activa fue del 13% (10,41% hipotiroidismo subclinico, 0,52% hipotiroidismo clinico, 1,56% hipertiroidismo subclinico y 0,52% hipertiroidismo clinico). La prevalencia de nuevos diagnosticos de EFT fue del 4,1% (7 hipotiroidismos y 1 hipertiroidismo, todos subclinicos). En el hipotiroidismo, se detectaron durante el estudio, las siguientes enfermedades: un 43% de trastornos de ansiedad, un 38% de sindromes depresivos, un 28,5% de deterioros cognitivos, un 9,5% de demencias, un 26% de alteraciones electrocardiograficas, un 47,6% de obesidad y un 28,5% de colesterol total . 250 mg/dl. En el hipertiroidismo se detectaron: un 50% de sindromes depresivos, un 25% de deterioros cognitivos, un 25% de alteraciones electrocardiograficas y un 50% de obesidadConclusionesLa prevalencia de EFT en la poblaciĂłn de edad avanzada es superior a la de la poblaciĂłn general, con un predominio del hipotiroidismo, las enfermedades subclĂnicas y las mujeres. Respecto a las enfermedades clásicamente asociadas a la disfunciĂłn tiroidea, se han observado pocas diferencias entre la poblaciĂłn afectada de EFT y la no afectada. El papel del mĂ©dico de atenciĂłn primaria es importante para disminuir el infradiagnĂłsticoObjectiveTo describe the prevalence of functional thyroid pathology (FTP) and pathologies associated with it in an elderly populationDesignDescriptive, cross-sectional studySettingUrban primary care centrePatientsRepresentative sample of the entire population attended that was 60 years old or overMain measurementsDemographic variables, clinicalhistory of thyroid pathology and pathology associated with it, Body Mass Index, small tests for diagnosing depression and anxiety, the mini-mental test, electrocardiogram, determination of total cholesterol and LDL cholesterol, and of free thyrotrophin and thyroxin if it is disturbedResults192 people were studied, 56% women, 53% between 60 and 69 and 12% over 79 years old. 10% had a history of previous FTP. Prevalence of active FTP was 13% (10.41% sub-clinical hypothyroidism, 0.52% clinical hypothyroidism, 1.56% subclinical hyperthyroidism, and 0.52% clinical hyperthyroidism). Prevalence of new diagnoses of FTP was 4.1% (7 with hypothyroidism and 1 with hyperthyroidism, all sub-clinical). During the study the following pathology was detected in hypothyroidism sufferers: 43% anxiety disorder, 38% depressive syndrome, 28.5% cognitive deterioration, 9.5% dementia, 26% electrocardiographic disturbances, 47.6% obesity, and 28.5% with total cholesterol .250 mg/dL. In hyperthyroidism, 50% with depressive syndrome, 25% with cognitive deterioration, 25% with electrocardiographic disturbances, and 50% with obesity were detectedConclusionsFTP is more prevalent among theelderly than in the population as a whole, with predominance of hypothyroidism, subclinical pathology and among women. In terms of pathology traditionally linked to thyroid malfunction, few differences were found between the population affectedwith FTP and those not affected. Primary care doctors are important in reducing underdiagnosi
Kids, Adolescents, and Young Adult Cancer Study—A Methodologic Approach in Cancer Epidemiology Research
Advances have been made in treatment and outcomes for pediatric cancer. However adolescents and young adults (AYAs) with cancer have not experienced similar relative improvements. We undertook a study to develop the methodology necessary for epidemiologic cancer research in these age groups. Our goal was to create the Kids, Adolescents, and Young Adults Cancer (KAYAC) project to create a resource to address research questions relevant to this population. We used a combination of clinic and population-based ascertainment to enroll 111 cases aged 0–39 for this methodology development study. The largest groups of cancer types enrolled include: breast cancer, leukemia, lymphoma, and melanoma. The overall participation rate is 69.8% and varies by age and tumor type. The study included patients, mothers, and fathers. The methods used to establish this resource are described, and the values of the resource in studies of childhood and young adult cancer are outlined
Transcriptional Profiling of the Caloric Restriction in Key Metabolic Tissues of Pigs Differing in Feed Efficiency
Residual feed intake is a measure of feed efficiency, where low RFI denotes high feed efficiency. Caloric restriction (CR) is associated with feed efficiency in livestock species and to human health benefits such as longevity and cancer prevention. We have developed pig lines that differ in RFI and we are interested in identifying the genes and pathways that underlie feed efficiency. Prepubertal Yorkshire gilts with low RFI (n=10) or high RFI (n=10) were fed ad libitum or at 80% of maintenance for 8 days. We measured serum metabolites and generated transcriptional profiles of liver and subcutaneous adipose tissue on these animals. Overall, 6,114 genes in fat and 305 genes in liver were differentially expressed (DE) in response to CR, and 311 genes in fat and 147 genes in liver were DE due to RFI differences. Pathway analyses of CR-induced DE genes indicated a dramatic switch to a conservation mode of energy usage by down-regulating lipogenesis and steroidogenesis in both liver and fat. Interestingly, CR altered expression of genes in immune and cell cycle/apoptotic pathways in fat, which may explain part of the CR-driven lifespan enhancement. In-silico analysis of transcription factors revealed ESR1 as a putative regulator of the adaptive response to CR, as several targets of ESR1 in our DE fat genes were annotated as cell cycle/apoptosis genes. The lipid metabolic pathway was overrepresented by down-regulated genes due to both CR and low RFI. We propose a common energy conservation mechanism, which may be controlled by PPARA, PPARG, and/or CREB in both CR and feed efficient pigs
Use of Transcriptional Profiling and Assessment of Blood Parameters to Understand Biological Mechanisms Controlling Feed Intake and Efficiency in Pigs
In this study, using transcriptional profiling of key tissues, we aimed to identify genetic mechanisms differing between control pigs and pigs that have been under selection for low residual feed intake (RFI) for three generations. A further aim was to determine the pathways responding to feed restriction within these lines and any line x treatment interactions resulting in gene expression differences. Preliminary results indicate that 2,809 genes in fat (p\u3c0.04, q\u3c0.2) and 61 genes in liver (p\u3c0.001, q\u3c0.2) showed differential expression in response to feed restriction. Also, 1,247 genes (p\u3c0.02, q\u3c0.2) showed differential expression between low RFI and control pigs and 38 genes (p\u3c0.001, q\u3c0.2) showed a line x feed interaction in liver. In addition, we measured the concentration of some of the important feed intake regulators in the blood such as leptin, triglyceride, and glucose. We found that the average blood leptin level to be significantly higher in the control ad libitum (CA) pigs than the control restricted (CR) group. Interestingly, the selected line of pigs on both restricted (SR) and ad libitum (SA) feed had similar blood leptin levels as found in the CR group pigs. Serum glucose levels were higher in CR than CA, however, we observed an opposite trend in the selected group. Combined with the transcriptional profiling results, blood hormone parameters may help us understand potential pathways that control FI and FE in pigs
Surgery Versus Radiotherapy for Clinically-localized Prostate Cancer: A Systematic Review and Meta-analysis
AbstractContextTo date, there is no Level 1 evidence comparing the efficacy of radical prostatectomy and radiotherapy for patients with clinically-localized prostate cancer.ObjectiveTo conduct a meta-analysis assessing the overall and prostate cancer-specific mortality among patients treated with radical prostatectomy or radiotherapy for clinically-localized prostate cancer.Evidence acquisitionWe searched Medline, EMBASE, and the Cochrane Library through June 2015 without year or language restriction, supplemented with hand search, using Preferred Reporting Items for Systematic Reviews and Meta-Analysis and Meta-analysis of Observational Studies in Epidemiology guidelines. We used multivariable adjusted hazard ratios (aHRs) to assess each endpoint. Risk of bias was assessed using the Newcastle-Ottawa scale.Evidence synthesisNineteen studies of low to moderate risk of bias were selected and up to 118 830 patients were pooled. Inclusion criteria and follow-up length varied between studies. Most studies assessed patients treated with external beam radiotherapy, although some included those treated with brachytherapy separately or with the external beam radiation therapy group. The risk of overall (10 studies, aHR 1.63, 95% confidence interval 1.54–1.73, p<0.00001; I2=0%) and prostate cancer-specific (15 studies, aHR 2.08, 95% confidence interval 1.76–2.47, p < 0.00001; I2=48%) mortality were higher for patients treated with radiotherapy compared with those treated with surgery. Subgroup analyses by risk group, radiation regimen, time period, and follow-up length did not alter the direction of results.ConclusionsRadiotherapy for prostate cancer is associated with an increased risk of overall and prostate cancer-specific mortality compared with surgery based on observational data with low to moderate risk of bias. These data, combined with the forthcoming randomized data, may aid clinical decision making.Patient summaryWe reviewed available studies assessing mortality after prostate cancer treatment with surgery or radiotherapy. While the studies used have a potential for bias due to their observational design, we demonstrated consistently higher mortality for patients treated with radiotherapy rather than surgery
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