Mutational analysis of BTAF1-TBP interaction: BTAF1 can rescue DNA-binding defective TBP mutants

The BTAF1 transcription factor interacts with TATA-binding protein (TBP) to form the B-TFIID complex, which is involved in RNA polymerase II transcription. Here, we present an extensive mapping study of TBP residues involved in BTAF1 interaction. This shows that residues in the concave, DNA-binding surface of TBP are important for BTAF1 binding. In addition, BTAF1 interacts with residues in helix 2 on the convex side of TBP as assayed in protein-protein and in DNA-binding assays. BTAF1 drastically changes the TATA-box binding specificity of TBP, as it is able to recruit DNA-binding defective TBP mutants to both TATA-containing and TATA-less DNA. Interestingly, other helix 2 interacting factors, such as TFIIA and NC2, can also stabilize mutant TBP binding to DNA. In contrast, TFIIB which interacts with a distinct surface of TBP does not display this activity. Since many proteins contact helix 2 of TBP, this provides a molecular basis for mutually exclusive TBP interactions and stresses the importance of this structural element for eukaryotic transcription

Medication adjustments in 1 out 3 patients as a result of pharmacogenetic testing initiated by primary care pharmacists based on specific inclusion protocols (SA-PGx pilot)

Medication adjustments in 7 out of 3 patients as a result of pharmacogenetic testing initiated by primary care pharmacists based on specific inclusion protocols (SA-PGx pilot) Background Pharmacogenetic analysis is currently performed at 16 certified laboratories in the Netherlands. For 90 different drugs pharmacogenetic advice is available, of which 23 drugs are frequently prescribed in primary care. Despite these opportunities the use of pharmacogenetic testing in primary care is still limited. Objective Investigate to what extent pharmacogenetic inclusion protocols and expert opinion of a pharmacist will lead to medication adjustments in patients. Design Prospective feasibility cohort study. Methods Eligible patients for pharmacogenetic testing are selected based on predefined inclusion protocols or pharmacist's expert opinion. DNA analysis is performed on buccal swab material collected in the pharmacy. Questionnaires, regarding drug-related complaints and patient satisfaction concerning provided care in the pharmacy, are filled in by the patients at baseline and after 1-2 weeks (no medication adjustments) or after 6 weeks (medication adjustments). Following study completion the application of pharmacogenetic testing is evaluated by the participating pharmacists. Results Over a period of 2 years 86 patients were included within 10 primary care pharmacies. Medication changes based on pharmacogenetic results were registered in 25 patients (29%). Almost 1/3 of these patients were selected based on the inclusion protocols. The most frequently changed drugs were clopidogrel, statins, pantoprazole and metoprolol. Drug complaints were reduced in 19% (n = 3) of the patients (not significant). The lack of imbursement in case of pharmacogenetic test requests from pharmacists was the most frequently mentioned barrier towards broad implementation of pharmacogenetics in primary care pharmacy. Conclusion Pharmacogenetic requests initiated by the pharmacist lead to medication adjustments in 30% of the patients with potentially also improvement in the pharmaco-therapy. Barriers such as imbursement require attention in order for pharmacogenetics to be applied more broadly in the primary care pharmacy.</p

Medication adjustments in 1 out 3 patients as a result of pharmacogenetic testing initiated by primary care pharmacists based on specific inclusion protocols (SA-PGx pilot)

Medication adjustments in 7 out of 3 patients as a result of pharmacogenetic testing initiated by primary care pharmacists based on specific inclusion protocols (SA-PGx pilot) Background Pharmacogenetic analysis is currently performed at 16 certified laboratories in the Netherlands. For 90 different drugs pharmacogenetic advice is available, of which 23 drugs are frequently prescribed in primary care. Despite these opportunities the use of pharmacogenetic testing in primary care is still limited. Objective Investigate to what extent pharmacogenetic inclusion protocols and expert opinion of a pharmacist will lead to medication adjustments in patients. Design Prospective feasibility cohort study. Methods Eligible patients for pharmacogenetic testing are selected based on predefined inclusion protocols or pharmacist's expert opinion. DNA analysis is performed on buccal swab material collected in the pharmacy. Questionnaires, regarding drug-related complaints and patient satisfaction concerning provided care in the pharmacy, are filled in by the patients at baseline and after 1-2 weeks (no medication adjustments) or after 6 weeks (medication adjustments). Following study completion the application of pharmacogenetic testing is evaluated by the participating pharmacists. Results Over a period of 2 years 86 patients were included within 10 primary care pharmacies. Medication changes based on pharmacogenetic results were registered in 25 patients (29%). Almost 1/3 of these patients were selected based on the inclusion protocols. The most frequently changed drugs were clopidogrel, statins, pantoprazole and metoprolol. Drug complaints were reduced in 19% (n = 3) of the patients (not significant). The lack of imbursement in case of pharmacogenetic test requests from pharmacists was the most frequently mentioned barrier towards broad implementation of pharmacogenetics in primary care pharmacy. Conclusion Pharmacogenetic requests initiated by the pharmacist lead to medication adjustments in 30% of the patients with potentially also improvement in the pharmaco-therapy. Barriers such as imbursement require attention in order for pharmacogenetics to be applied more broadly in the primary care pharmacy.</p

Short- and Long-Term Biomarkers for Bacterial Robustness: A Framework for Quantifying Correlations between Cellular Indicators and Adaptive Behavior

The ability of microorganisms to adapt to changing environments challenges the prediction of their history-dependent behavior. Cellular biomarkers that are quantitatively correlated to stress adaptive behavior will facilitate our ability to predict the impact of these adaptive traits. Here, we present a framework for identifying cellular biomarkers for mild stress induced enhanced microbial robustness towards lethal stresses. Several candidate-biomarkers were selected by comparing the genome-wide transcriptome profiles of our model-organism Bacillus cereus upon exposure to four mild stress conditions (mild heat, acid, salt and oxidative stress). These candidate-biomarkers—a transcriptional regulator (activating general stress responses), enzymes (removing reactive oxygen species), and chaperones and proteases (maintaining protein quality)—were quantitatively determined at transcript, protein and/or activity level upon exposure to mild heat, acid, salt and oxidative stress for various time intervals. Both unstressed and mild stress treated cells were also exposed to lethal stress conditions (severe heat, acid and oxidative stress) to quantify the robustness advantage provided by mild stress pretreatment. To evaluate whether the candidate-biomarkers could predict the robustness enhancement towards lethal stress elicited by mild stress pretreatment, the biomarker responses upon mild stress treatment were correlated to mild stress induced robustness towards lethal stress. Both short- and long-term biomarkers could be identified of which their induction levels were correlated to mild stress induced enhanced robustness towards lethal heat, acid and/or oxidative stress, respectively, and are therefore predictive cellular indicators for mild stress induced enhanced robustness. The identified biomarkers are among the most consistently induced cellular components in stress responses and ubiquitous in biology, supporting extrapolation to other microorganisms than B. cereus. Our quantitative, systematic approach provides a framework to search for these biomarkers and to evaluate their predictive quality in order to select promising biomarkers that can serve to early detect and predict adaptive traits

Variations in activin receptor, inhibin/activin subunit and follistatin mRNAs in human prostate tumour tissues

The possible role of activin in the regulation of malignant prostatic growth was studied using RNAase protection assays of activin receptors, inhibin/activin subunits and follistatin mRNAs in the human prostatic carcinoma cell lines LNCaP-FGC, -R and -LNO, in human prostatic carcinoma xenografts and in human prostatic tissue. Activin receptor types IA (ActRIA), IB (ActRIB), IIA (ActRIIA) and IIB (ActRIIB) mRNAs were generally expressed in prostate pithelial cells, with significantly lower levels of ActRIB mRNA in prostate tumour aterial when compared to non-malignant tissue (P< 0.05; Mann–Whitney U -test). Inhibin/activin βA- and βB-subunit mRNA expression was also found in prostate tissue. Androgen-independent xenografts expressed significantly lower amounts of βB-subunit mRNA when compared to androgen-dependent xenografts (P< 0.05). While βB-subunit mRNA was expressed by LNCaP-FGC and -LNO cells, virtually no expression was found in the androgen-independent LNCaP-R line. Inhibin α-subunit mRNA levels were low or undetectable in all samples investigated. Follistatin mRNA was undetectable in LNCaP-sublines, while low levels were found in prostatic tissues. In androgen-independent LNCaP-R cells, activin inhibited cell growth in a dose-dependent manner. These results suggest that prostate tumour progression is accompanied by a decrease of the inhibitory effect of locally produced activin by either a decrease in the expression of activin βB-subunit mRNA or by a decrease of ActRIB mRNA levels. © 2000 Cancer Research Campaig

Resting energy expenditure in children at risk of hypothalamic dysfunction

Objective: Children with suprasellar brain damage are at risk of hypothalamic dysfunction (HD). HD may lead to decreased resting energy expenditure (REE). Decreased REE, however, is not present in all children with HD. Our aim was to assess which children suspect for HD have low REE, and its association with clinical severity of HD or radiological hypothalamic damage. Patients and methods: A retrospective cohort study was performed. Measured REE (mREE) of children at risk of HD was compared to predicted REE (pREE). Low REE was defined as mREE <90% of predicted. The mREE/pREE quotient was associated to a clinical score for HD symptoms and to radiological hypothalamic damage. Results: In total, 67 children at risk of HD (96% brain tumor diagnosis) with a mean BMI SDS of +2.3 ± 1.0 were included. Of these, 45 (67.2%) had low mREE. Children with severe HD had a significant lower mean mREE/pREE quotient compared to children with no, mild, or moderate HD. Mean mREE/pREE quotient of children with posterior hypothalamic damage was significantly lower compared to children with no or anterior damage. Tumor progression or tumor recurrence, severe clinical HD, and panhypopituitarism with diabetes insipidus (DI) were significant risk factors for reduced REE. Conclusion: REE may be lowered in children with hypothalamic damage and is associated to the degree of clinical HD. REE is, however, not lowered in all children suspect for HD. For children with mild or moderate clinical HD symptoms, REE measurements may be useful to distinguish between those who may benefit from obesity treatment that increases REE from those who would be better helped using other obesity interventions

REVISITING ANNA MOSCOWITZ\u27S KROSS\u27S CRITIQUE OF NEW YORK CITY\u27S WOMEN\u27S COURT: THE CONTINUED PROBLEM OF SOLVING THE PROBLEM OF PROSTITUTION WITH SPECIALIZED CRIMINAL COURTS

This article explores New York City\u27s non-traditional, judicially based response to prostitution. This article first recounts the history of New York City’s Women’s Court. It then examines the work of the Midtown Community Court, the “problem-solving court” established in 1993 to address criminal issues, like prostitution, in Midtown Manhattan. It also discusses the renewed concerns about sex work in New York and describe the movement, propelled by modern reformers, to address prostitution through specialty courts. It then contrasts the shared features and attributes of the Women’s Court and Midtown Court models. Finally, the article urges modern reformers to step back from the problem-solving court movement and their call for the creation of more such specialized criminal courts