Identifying gene locus associations with promyelocytic leukemia nuclear bodies using immuno-TRAP.

Important insights into nuclear function would arise if gene loci physically interacting with particular subnuclear domains could be readily identified. Immunofluorescence microscopy combined with fluorescence in situ hybridization (immuno-FISH), the method that would typically be used in such a study, is limited by spatial resolution and requires prior assumptions for selecting genes to probe. Our new technique, immuno-TRAP, overcomes these limitations. Using promyelocytic leukemia nuclear bodies (PML NBs) as a model, we used immuno-TRAP to determine if specific genes localize within molecular dimensions with these bodies. Although we confirmed a TP53 gene-PML NB association, immuno-TRAP allowed us to uncover novel locus-PML NB associations, including the ABCA7 and TFF1 loci and, most surprisingly, the PML locus itself. These associations were cell type specific and reflected the cell's physiological state. Combined with microarrays or deep sequencing, immuno-TRAP provides powerful opportunities for identifying gene locus associations with potentially any nuclear subcompartment

Medication-induced Uveitis: An Update

Drug-induced uveitis is an uncommon but important cause of ocular inflammation. Uveitis can be seen in association with various systemic, topical, and intraocular medications. In this article, we review common medications associated with uveitis. Most cases of drug-induced uveitis resolve with termination of the suspected medication with or without administration of topical or systemic steroids. It is important for clinicians to readily identify medications that may cause uveitis in order to provide rapid treatment, avoid consequences of longstanding inflammation, and prevent costly and excessive laboratory testing

First-order symmetrizable hyperbolic formulations of Einstein's equations including lapse and shift as dynamical fields

First-order hyperbolic systems are promising as a basis for numerical integration of Einstein's equations. In previous work, the lapse and shift have typically not been considered part of the hyperbolic system and have been prescribed independently. This can be expensive computationally, especially if the prescription involves solving elliptic equations. Therefore, including the lapse and shift in the hyperbolic system could be advantageous for numerical work. In this paper, two first-order symmetrizable hyperbolic systems are presented that include the lapse and shift as dynamical fields and have only physical characteristic speeds.Comment: 11 page

Motivation for Choosing Teaching as a Career and Job Satisfaction with Context of Pakistan Administrative Kashmir

This study examines the career motivation and job satisfaction of 150 public primary school teachers of Pakistan administrative Kashmir who have chosen teaching as a career. So, they were asked questions about factors influencing to choosing teaching as career perceptions, major expectations and five factors of job satisfaction which include work, promotion, salary, co-worker and supervision. A profile of the participants was then developed by analyzing their responses in quantitative way as being descriptive, statistical, and inductive steps. It has been examined that both extrinsic and intrinsic motivations play a role when individuals choose teaching as a career but most of the primary teachers choose teaching career for intrinsic reasons such as they always wanted to become a teacher as they wanted to do something for nation through this profession. But they are moderate in term of job satisfaction. Keywords: Motivation, Teaching Career, Job satisfaction, Primary Teachers and AJ&

Experimental study of multi-pass solar air thermal collector system assisted with sensible energy-storing matrix

Analysis of heat transfer around the components of multi-pass solar air heating collector and its per�formance enhancement has been presented using experimental approach. Radiation and convection heat transfer from the system covers and collector plate to the flowing airstreams in the collector system have been investigated. Geological porous matrix has been used as thermal energy reservoir. Test rig was set up under daily average solar insolation, ambient temperature, wind speed, and relative humidity of 302.40 Wm-2, 29.30 �C, 0.58 ms�1 , and 48.53%, respectively. Thermal performance evaluation of the multi-pass solar air collector has been conducted in accordance with the ASHRAE standard specified for the solar air heating systems. Thermal collector efficiency range of 51.91e72.55% has been achieved with an optimum air mass flow rate of 0.013 kgs�1 . System evaporative capacity range of 1:158 � 10�3 - 1:205 � 10�3 kgs�1 was computed. The matrix has extended the span of system operation for more than 3 h after sunset. Comparison of the outcome of multi-pass solar collector performance with the reported study has been done with good agreement. However, an improvement in performance through heat transfer from collector to the flowing air could be achieved

Calpain 1 Knockdown Improves Tissue Sparing and Functional Outcomes after Spinal Cord Injury in Rats

To evaluate the hypothesis that calpain 1 knockdown would reduce pathological damage and functional deficits after spinal cord injury (SCI), we developed lentiviral vectors encoding calpain 1 shRNA and eGFP as a reporter (LV-CAPN1 shRNA). The ability of LV-CAPN1 shRNA to knockdown calpain 1 was confirmed in rat NRK cells using Northern and Western blot analysis. To investigate the effects on spinal cord injury, LV-CAPN1shRNA or LV-mismatch control shRNA (LV-control shRNA) were administered by convection enhanced diffusion at spinal cord level T10 in Long-Evans female rats (200–250 g) 1 week before contusion SCI, 180 kdyn force, or sham surgery at the same thoracic level. Intraspinal administration of the lentiviral particles resulted in transgene expression, visualized by eGFP, in spinal tissue at 2 weeks after infection. Calpain 1 protein levels were reduced by 54% at T10 2 weeks after shRNA-mediated knockdown (p\u3c0.05, compared with the LV-control group, n=3 per group) while calpain 2 levels were unchanged. Intraspinal administration of LV-CAPN1shRNA 1 week before contusion SCI resulted in a significant improvement in locomotor function over 6 weeks postinjury, compared with LV-control administration (p\u3c0.05, n=10 per group). Histological analysis of spinal cord sections indicated that pre-injury intraspinal administration of LV-CAPN1shRNA significantly reduced spinal lesion volume and improved total tissue sparing, white matter sparing, and gray matter sparing (p\u3c0.05, n=10 per group). Together, results support the hypothesis that calpain 1 activation contributes to the tissue damage and impaired locomotor function after SCI, and that calpain1 represents a potential therapeutic target

General practitioners\u27 knowledge and approach to chronic kidney disease in Karachi, Pakistan

Due to lack of adequate number of formally trained nephrologists, many patients with chronic kidney disease (CKD) are seen by general practitioners (GPs). This study was designed to assess the knowledge of the GPs regarding identification of CKD and its risk factors, and evaluation and management of risk factors as well as complications of CKD. We conducted a cross-sectional survey of 232 randomly selected GPs from Karachi during 2011. Data were collected on a structured questionnaire based on the kidney disease outcomes and quality initiative recommendations on screening, diagnosis, and management of CKD. A total of 235 GPs were approached, and 232 consented to participate. Mean age was 38.5 ± 11.26 years; 56.5% were men. Most of the GPs knew the traditional risk factors for CKD, i.e., diabetes (88.4%) and hypertension (80%), but were less aware of other risk factors. Only 38% GPs were aware of estimated glomerular filtration rate in evaluation of patients with CKD. Only 61.6% GPs recognized CKD as a risk factor for cardiovascular disease. About 40% and 29% GPs knew the correct goal systolic and diastolic blood pressure, respectively. In all, 41% GPs did not know when to refer the patient to a nephrologist. Our survey identified specific gaps in knowledge and approach of GPs regarding diagnosis and management of CKD. Educational efforts are needed to increase awareness of clinical practice guidelines and recommendations for patients with CKD among GPs, which may improve management and clinical outcomes of this population

Performance of high-throughput DNA quantification methods

BACKGROUND: The accuracy and precision of estimates of DNA concentration are critical factors for efficient use of DNA samples in high-throughput genotype and sequence analyses. We evaluated the performance of spectrophotometric (OD) DNA quantification, and compared it to two fluorometric quantification methods, the PicoGreen(® )assay (PG), and a novel real-time quantitative genomic PCR assay (QG) specific to a region at the human BRCA1 locus. Twenty-Two lymphoblastoid cell line DNA samples with an initial concentration of ~350 ng/uL were diluted to 20 ng/uL. DNA concentration was estimated by OD and further diluted to 5 ng/uL. The concentrations of multiple aliquots of the final dilution were measured by the OD, QG and PG methods. The effects of manual and robotic laboratory sample handling procedures on the estimates of DNA concentration were assessed using variance components analyses. RESULTS: The OD method was the DNA quantification method most concordant with the reference sample among the three methods evaluated. A large fraction of the total variance for all three methods (36.0–95.7%) was explained by sample-to-sample variation, whereas the amount of variance attributable to sample handling was small (0.8–17.5%). Residual error (3.2–59.4%), corresponding to un-modelled factors, contributed a greater extent to the total variation than the sample handling procedures. CONCLUSION: The application of a specific DNA quantification method to a particular molecular genetic laboratory protocol must take into account the accuracy and precision of the specific method, as well as the requirements of the experimental workflow with respect to sample volumes and throughput. While OD was the most concordant and precise DNA quantification method in this study, the information provided by the quantitative PCR assay regarding the suitability of DNA samples for PCR may be an essential factor for some protocols, despite the decreased concordance and precision of this method