Alternative splicing affects the function and tissue-specific expression of the human constitutive androstane receptor

BACKGROUND: The constitutive androstane receptor (CAR) plays a key role in the control of drug metabolism and transport by mediating the phenobarbital-type induction of many phase I and II drug metabolizing enzymes and drug transporters. RESULTS: We identified transcripts generated by four different alternative splicing events in the human CAR gene. Two of the corresponding ligand binding domain isoforms demonstrated novel functional properties: First, CAR(SV3), which is encoded by a transcript containing an lengthened exon 7, differentially transactivated target gene promoters. Second, CAR(SV2), which results from the use of an alternative 3' splice site lengthening exon 8, showed ligand-dependent instead of constitutive interaction with coactivators. Furthermore, alternatively spliced transcripts demonstrated a tissue-specific expression pattern. In most tissues, only transcripts generated by alternative splicing within exon 9 were expressed. The encoded variant demonstrated a loss-of-function phenotype. Correct splicing of exon 8 to exon 9 is restricted to only a few tissues, among them liver and small intestine for which CAR function has been demonstrated, and is associated with the induction of CAR expression during differentiation of intestinal cells. CONCLUSION: Due to their specific activities, CAR variant proteins SV2 and SV3 may modulate the activity of reference CAR(SV1). Furthermore, we propose that transcriptional activation and regulation of splicing of exon 9 may be coupled to ensure appropriate tissue- and differentiation state-specific expression of transcripts encoding functional CAR protein. Altogether, alternative splicing seems to be of utmost importance for the regulation of CAR expression and function

Association between obesity, quality of life, physical activity and health service utilization in primary care patients with osteoarthritis

OBJECTIVE: To assess the association of obesity with quality of life, health service utilization and physical activity in a large sample of primary care patients with osteoarthritis (OA). METHODS: Data were retrieved from the PraxArt project, representing a cohort of 1021 primary care patients with OA. In 978 patients, height and weight were measured and the Body Mass Index (BMI) was calculated. The AIMS2-SF was used to assess quality of life (QoL). Data about health service utilization (HSU) were retrieved by means of patients' medical files. Concomitant depression was assessed by means of the Patient Health Questionnaire (PHQ-9). Patients were grouped into normal weight, overweight and obese according to the definition of the WHO and compared by means of analysis of covariance (ANCOVA). RESULTS: Obese and overweight persons achieved significantly higher scores on the AIMS2-SF lower body scale, the symptom, the affect and the work scale, indicating an increased burden by OA. The PHQ-9 score increased significantly over the three weight-groups, indicating a positive association of BMI and depression. With increasing BMI, the number of comorbidities increased and physical activity decreased significantly. After controlling for covariates, contacts to orthopaedics and performed x-rays remained significantly higher in obese patients, but not contacts to general practitioners. CONCLUSION: The results display a strong association of QoL and BMI, resulting in increased use of the health care system. Thus, the study emphasizes the need for appropriate approaches in primary care to break the vicious circle of overweight, depression, decreasing physical inactivity and decreasing QoL

Epidemiologic, Phenotypic, and Structural Characterization of Aminoglycoside-Resistance Gene aac(3)-IV

Aminoglycoside antibiotics are powerful bactericidal therapeutics that are often used in the treatment of critical Gram-negative systemic infections. The emergence and global spread of antibiotic resistance, however, has compromised the clinical utility of aminoglycosides to an extent similar to that found for all other antibiotic-drug classes. Apramycin, a drug candidate currently in clinical development, was suggested as a next-generation aminoglycoside antibiotic with minimal cross-resistance to all other standard-of-care aminoglycosides. Here, we analyzed 591,140 pathogen genomes deposited in the NCBI National Database of Antibiotic Resistant Organisms (NDARO) for annotations of apramycin-resistance genes, and compared them to the genotypic prevalence of carbapenem resistance and 16S-rRNA methyltransferase (RMTase) genes. The 3-N-acetyltransferase gene aac(3)-IV was found to be the only apramycin-resistance gene of clinical relevance, at an average prevalence of 0.7%, which was four-fold lower than that of RMTase genes. In the important subpopulation of carbapenemase-positive isolates, aac(3)-IV was nine-fold less prevalent than RMTase genes. The phenotypic profiling of selected clinical isolates and recombinant strains expressing the aac(3)-IV gene confirmed resistance to not only apramycin, but also gentamicin, tobramycin, and paromomycin. Probing the structure-activity relationship of such substrate promiscuity by site-directed mutagenesis of the aminoglycoside-binding pocket in the acetyltransferase AAC(3)-IV revealed the molecular contacts to His124, Glu185, and Asp187 to be equally critical in binding to apramycin and gentamicin, whereas Asp67 was found to be a discriminating contact. Our findings suggest that aminoglycoside cross-resistance to apramycin in clinical isolates is limited to the substrate promiscuity of a single gene, rendering apramycin best-in-class for the coverage of carbapenem- and aminoglycoside-resistant bacterial infections

Is the job satisfaction of primary care team members associated with patient satisfaction?

Item does not contain fulltextBACKGROUND Previous research has shown a correlation between physician job satisfaction and patient satisfaction with quality of care, but the connection between job satisfaction of other primary care team members and patient satisfaction is yet unclear. OBJECTIVE To evaluate whether there is an association between patient satisfaction and job satisfaction of the members of patient care teams. DESIGN The study was based on data from the European Practice Assessment and used an observational design. SETTING 676 primary care practices in Germany. PARTICIPANTS 47 168 patients, 676 general practitioners (practice principals), 305 physician colleagues (trainees and permanently employed physicians) and 3011 non-physician practice members (nurses, secretaries). MAIN OUTCOME MEASURES Patient evaluation was measured using the 23-item EUROPEP questionnaire. Job satisfaction was measured using the 10-item Warr-Cook-Wall job satisfaction scale and further items relating to practice structure. Bivariate correlations were applied in which factors of patient satisfaction and practice structure were compared with physicians and non-physicians satisfaction. RESULTS Patient satisfaction correlates positively with the general job satisfaction of the non-physician (r=0.25, p<0.01) and no significant correlation was found for the general job satisfaction of practice principals and physician colleagues. Patients' satisfaction with the practice organisation correlates positively with the general job satisfaction of the non-physicians (r=0.30, p<0.01) and their view of practice structure (r=0.29, p<0.01). CONCLUSIONS The correlation between non-physician team member satisfaction and patient satisfaction was higher than the correlation between satisfaction of physicians and patients. Patients seem to be sensitive to aspects of practice structure.1 juni 201

Translation and validation of the PACIC+ questionnaire: the Thai version

Background: The number of patients with chronic illness is increasing worldwide. These patients usually receive care from a primary care facility. The Patient Assessment of Chronic Illness Care (PACIC) is a tool that is increasingly used in several countries to measure how the patients perceive the care they receive. The goal of this validation study is to provide and validate an extended version of the tool, the PACIC+ questionnaire, in Thailand. Methods: In this observational validation study, patients with type 2 diabetes from the outpatient clinic at a university hospital in Thailand completed the PACIC+ at the clinic. For follow-up, they received the questionnaire per mail after four weeks. The Thai PACIC+ comprises 26 items, which map onto 5 subscales and a summary score related to the Chronic Care Model (CCM) and 5 subscales and a summary score related to the 5A model, a counseling model for behavioral changes. Data-analysis focused on the use of most extreme answering categories (&gt; 15%), internal consistency (Cronbach’s alpha), and test-retest reliability. An exploratory factor analysis (EFA) was performed for the CCM and the 5A model separately to examine the factor structure. Results: A total of 151 patients participated. The average age of the sample was 63 ± 9 years (range 29–86 years). Fifty-three percent of the respondents were female. In the Delivery System subscale, 20% of patients reported the highest possible value; in all other subscales, relative frequencies of the most extreme categories did not exceed 15%. Cronbach’s alpha per subscale varied from 0.58 to 0.81, while that of the summary scores were 0.89 and 0.91. The mean difference from the test-retest varied from − 0.06 to 0.17 across subscales. The Kaiser-Meyer-Olkin criterion for sampling adequacy (KMO) was good for both models as well as the Bartlett’s test for sphericity p. While the factor loadings in rotated factor solution showed good concordance with the CCM, concordance was not as good for the 5A model, especially for the subscales “Assess” and “Advice”. Conclusion: A validated Thai version of the PACIC+ is now available to measure how the patients perceive the care they receive

Novel oral anticoagulants in primary care in patients with atrial fibrillation: a cross-sectional comparison before and after their introduction

Background: Novel oral anticoagulation (NOAC) has been introduced in recent years, but data on use in atrial fibrillation (AF) in primary care setting is scarce. In Germany, General Practitioners are free to choose type of oral anticoagulation (OAC) in AF. Our aim was to explore changes in prescription-rates of OAC in German primary care before and after introduction of NOAC on the market. Methods: Data of a representative morbidity registration project in primary care in Germany (CONTENT) were analysed. Patients with AF in 2011 or 2014 were included (before and after broad market authorization of NOAC, respectively). We defined three independent groups: patients from 2011 without follow-up (group A), patients from 2014 but without previous record in 2011 (group B) and patients with AF and records in 2011 and 2014 (group C). Results: 2642 patients were included. Group A (n = 804) and B (n = 755) were comparable regarding patient characteristics. 87.3% of group A and 84.8% of group B had CHA2DS2-VASc-Score ≥ 2, indicating a need for oral anticoagulation (OAC). Prescription of OAC increased from 23.1% (n = 186) to 42.8% (n = 323, p &lt; .01) with stable use of vitamin-k-antagonist (22.6–24.9%). NOAC increased from 0.6 to 19.2% (p &lt; .01). Monotherapy with Acetylsalicylic acid (ASA) decreased from 15.3% (n = 123) to 8.2% (n = 62, p &lt; .01). In group C (n = 1083), OAC increased from 35.3 to 55.4% (p &lt; .01), with stable prescription rate of vitamin-k-antagonist (34.4–35.7%). NOAC increased from 0.9 to 21.5% (p &lt; .01). Conclusions: In summary, our study showed a significant increase of OAC over time, which is fostered by the use of NOAC but with a stable rate of VKA and a sharp decrease of ASA. Patients on VKA are rarely switched to NOAC, but new patients with AF are more likely to receive NOAC

Present climate and climate change over North America as simulated by the fifth-generation Canadian regional climate model

The fifth-generation Canadian Regional Climate Model (CRCM5) was used to dynamically downscale two Coupled Global Climate Model (CGCM) simulations of the transient climate change for the period 1950–2100, over North America, following the CORDEX protocol. The CRCM5 was driven by data from the CanESM2 and MPI-ESM-LR CGCM simulations, based on the historical (1850–2005) and future (2006–2100) RCP4.5 radiative forcing scenario. The results show that the CRCM5 simulations reproduce relatively well the current-climate North American regional climatic features, such as the temperature and precipitation multiannual means, annual cycles and temporal variability at daily scale. A cold bias was noted during the winter season over western and southern portions of the continent. CRCM5-simulated precipitation accumulations at daily temporal scale are much more realistic when compared with its driving CGCM simulations, especially in summer when small-scale driven convective precipitation has a large contribution over land. The CRCM5 climate projections imply a general warming over the continent in the 21st century, especially over the northern regions in winter. The winter warming is mostly contributed by the lower percentiles of daily temperatures, implying a reduction in the frequency and intensity of cold waves. A precipitation decrease is projected over Central America and an increase over the rest of the continent. For the average precipitation change in summer however there is little consensus between the simulations. Some of these differences can be attributed to the uncertainties in CGCM-projected changes in the position and strength of the Pacific Ocean subtropical high pressure

Mycobacterial arthritis and synovitis in painted reed frogs (Hyperolius marmoratus)

Several species of atypical mycobacteria have been isolated from wild and captive amphibians. In captive anurans, cutaneous and visceral mycobacteriosis are common and can result in significant mortality, particularly when animals are immunocompromised. Mycobacterial arthritis and synovitis are reported rarely in amphibians. We describe 20 cases in painted reed frogs (Hyperolius marmoratus), which presented with cachexia, limb paresis or paralysis or ‘spindly leg syndrome’. Histopathology revealed multifocal histiocytic to granulomatous synovitis affecting appendicular, rib or spinal intervertebral joints. Periarticular granulomata, granulomatous cellulitis and skeletal muscle atrophy, necrosis and degeneration were also present. In one case, granulomatous spinal osteomyelitis was recorded. ZiehleNeelsen stains showed large numbers of acid-fast bacteria in macrophages and histiocytes. The mycobacterial isolates obtained from culture were identified as members of the Mycobacterium chelonae complex (either M. chelonae or Mycobacterium abscessus). This was confirmed by 50 -16S ribosomal ribonucleic acid (rRNA) sequencing. In 17 cases mycobacterial lesions were present only in the joints and skeleton, highlighting the importance of not ruling out mycobacterial infection on the basis of absence of cutaneous or visceral lesions.http://elsevier.com/locate/jcpa2018-02-28hb2017Production Animal Studie