Regulation of phenylacetic acid uptake is σ54 dependent in Pseudomonas putida CA-3

<p>Abstract</p> <p>Background</p> <p>Styrene is a toxic and potentially carcinogenic alkenylbenzene used extensively in the polymer processing industry. Significant quantities of contaminated liquid waste are generated annually as a consequence. However, styrene is not a true xenobiotic and microbial pathways for its aerobic assimilation, via an intermediate, phenylacetic acid, have been identified in a diverse range of environmental isolates. The potential for microbial bioremediation of styrene waste has received considerable research attention over the last number of years. As a result the structure, organisation and encoded function of the genes responsible for styrene and phenylacetic acid sensing, uptake and catabolism have been elucidated. However, a limited understanding persists in relation to host specific regulatory molecules which may impart additional control over these pathways. In this study the styrene degrader <it>Pseudomonas putida </it>CA-3 was subjected to random mini-Tn<it>5 </it>mutagenesis and mutants screened for altered styrene/phenylacetic acid utilisation profiles potentially linked to non-catabolon encoded regulatory influences.</p> <p>Results</p> <p>One mutant, D7, capable of growth on styrene, but not on phenylacetic acid, harboured a Tn<it>5 </it>insertion in the <it>rpoN </it>gene encoding σ54. Complementation of the D7 mutant with the wild type <it>rpoN </it>gene restored the ability of this strain to utilise phenylacetic acid as a sole carbon source. Subsequent RT-PCR analyses revealed that a phenylacetate permease, PaaL, was expressed in wild type <it>P. putida </it>CA-3 cells utilising styrene or phenylacetic acid, but could not be detected in the disrupted D7 mutant. Expression of plasmid borne <it>paaL </it>in mutant D7 was found to fully restore the phenylacetic acid utilisation capacity of the strain to wild type levels. Bioinformatic analysis of the <it>paaL </it>promoter from <it>P. putida </it>CA-3 revealed two σ⁵⁴consensus binding sites in a non-archetypal configuration, with the transcriptional start site being resolved by primer extension analysis. Comparative analyses of genomes encoding phenylacetyl CoA, (PACoA), catabolic operons identified a common association among styrene degradation linked PACoA catabolons in <it>Pseudomonas </it>species studied to date.</p> <p>Conclusions</p> <p>In summary, this is the first study to report RpoN dependent transcriptional activation of the PACoA catabolon <it>paaL </it>gene, encoding a transport protein essential for phenylacetic acid utilisation in <it>P. putida </it>CA-3. Bioinformatic analysis is provided to suggest this regulatory link may be common among styrene degrading <it>Pseudomonads</it>.</p

Bevacizumab for newly diagnosed ovarian cancers: Best candidates among high-risk disease patients (icon-7)

Bevacizumab is approved as a maintenance treatment in first-line setting in advanced-stage III-IV ovarian cancers, because GOG-0218 and ICON-7 phase III trials demonstrated progression-free survival benefits. However, only the subgroup of patients with high-risk diseases (stage IV, and incompletely resected stage III) derived an overall survival (OS) gain in the ICON-7 trial (4.8 months). The modeled CA-125 elimination rate constant K (KELIM) parameter, based on the longitudinal CA- 125 kinetics during the first 100 days of chemotherapy, is a potential indicator of the tumor primary chemo-sensitivity. In the ICON-7 trial dataset, the OS of patients within the low- and high-risk disease groups was assessed according to treatment arms and KELIM. Among the patients with high-risk diseases, those with favorable standardized KELIM of at least 1.0 (n=214, 46.7%) had no survival benefit from bevacizumab, whereas those with unfavorable KELIM less than 1.0 (n=244, 53.2%) derived the highest OS benefit (absolute difference = 9.1 months, 2-sided log-rank P=.10; Cox hazard ratio = 0.78, 95% confidence interval = 0.58 to 1.04, 2-sided P=.09)

Bevacizumab for Newly Diagnosed Ovarian Cancers: Best Candidates Among High-Risk Disease Patients (ICON-7)

Bevacizumab is approved as a maintenance treatment in first-line setting in advanced-stage III-IV ovarian cancers, because GOG-0218 and ICON-7 phase III trials demonstrated progression-free survival benefits. However, only the subgroup of patients with high-risk diseases (stage IV, and incompletely resected stage III) derived an overall survival (OS) gain in the ICON-7 trial (4.8 months). The modeled CA-125 elimination rate constant K (KELIM) parameter, based on the longitudinal CA-125 kinetics during the first 100 days of chemotherapy, is a potential indicator of the tumor primary chemo-sensitivity. In the ICON-7 trial dataset, the OS of patients within the low- and high-risk disease groups was assessed according to treatment arms and KELIM. Among the patients with high-risk diseases, those with favorable standardized KELIM of at least 1.0 (n = 214, 46.7%) had no survival benefit from bevacizumab, whereas those with unfavorable KELIM less than 1.0 (n = 244, 53.2%) derived the highest OS benefit (absolute difference = 9.1 months, 2-sided log-rank P = .10; Cox hazard ratio = 0.78, 95% confidence interval = 0.58 to 1.04, 2-sided P = .09)

Unbiased Proteomic Approach Identifies Pathobiological Profiles in the Brains of Preclinical Models of Repetitive Mild Traumatic Brain Injury, Tauopathy, and Amyloidosis

No concerted investigation has been conducted to explore overlapping and distinct pathobiological mechanisms between repetitive mild traumatic brain injury (r-mTBI) and tau/amyloid proteinopathies considering the long history of association between TBI and Alzheimer’s disease. We address this problem by using unbiased proteomic approaches to generate detailed time-dependent brain molecular profiles of response to repetitive mTBI in C57BL/6 mice and in mouse models of amyloidosis (with amyloid precursor protein KM670/671NL (Swedish) and Presenilin 1 M146L mutations [PSAPP]) and tauopathy (hTau). Brain tissues from animals were collected at different timepoints after injuries (24 hr–12 months post-injury) and at different ages for tau or amyloid transgenic models (3, 9, and 15 months old), encompassing the pre-, peri-, and post-“onset” of cognitive and pathological phenotypes. We identified 30 hippocampal and 47 cortical proteins that were significantly modulated over time in the r-mTBI compared with sham mice. These proteins identified TBI-dependent modulation of phosphatidylinositol-3-kinase/AKT signaling, protein kinase A signaling, and PPARα/RXRα activation in the hippocampus and protein kinase A signaling, gonadotropin-releasing hormone signaling, and B cell receptor signaling in the cortex. Previously published neuropathological studies of our mTBI model showed a lack of amyloid and tau pathology. In PSAPP mice, we identified 19 proteins significantly changing in the cortex and only 7 proteins in hTau mice versus wild-type littermates. When we explored the overlap between our r-mTBI model and the PSAPP/hTau models, a fairly small coincidental change was observed involving only eight significantly regulated proteins. This work suggests a very distinct TBI neurodegeneration and also that other factors are needed to drive pathologies such as amyloidosis and tauopathy postinjury

Squamous cell carcinoma of the nasal cavity:A descriptive analysis of cases from the Head and Neck 5000 study

OBJECTIVES: This paper aims to provide contemporary epidemiological data on squamous cell carcinoma (SCC) of the nasal cavity, which represents a rare type of head and neck cancer.DESIGN, SETTING &amp; PARTICIPANTS: A descriptive analysis of people with nasal cavity SCC treated with curative intent from the Head and Neck 5000 study; a multicentre clinical cohort study of people from the UK with head and neck cancer. People with tumours of the nasopharynx, paranasal sinuses and other sub-sites of the head and neck were excluded.MAIN OUTCOME MEASURES: Demographic data and treatment details are presented for all participants. The main outcomes were overall survival and survival according to categories of characteristics (e.g. smoker vs non-smoker); these were explored using Kaplan-Meier plots.RESULTS: Thirty people with nasal cavity SCC were included in the study, of which most were male (67%) and current or ex-smokers (70%). The majority (70%) presented with early stage (T1/2, N0) tumours. Cervical lymph node metastases at presentation were rare, occurring in only one person. Nine people died during the follow up period (30%). Worse survival outcomes were seen in people with moderate or severe co-morbidities.CONCLUSIONS: This paper provides epidemiological data on nasal cavity SCC in the UK. Patterns of disease and survival outcomes are described, identifying high-risk groups. Further studies should explore whether primary treatment modality alters survival. This article is protected by copyright. All rights reserved.</p

The prescribing of prisms in clinical practice

The use of prisms in cases of decompensated heterophoria is an established treatment modality. The clinical literature lacks consensus upon the appropriate use of prisms, and fails to provide the necessary evidence base. While the experimental literature can guide the practitioner, the lack of double-blind, placebo-controlled clinical studies needs to be addressed

Thermodynamics of C incorporation on Si(100) from ab initio calculations

We study the thermodynamics of C incorporation on Si(100), a system where strain and chemical effects are both important. Our analysis is based on first-principles atomistic calculations to obtain the important lowest energy structures, and a classical effective Hamiltonian which is employed to represent the long-range strain effects and incorporate the thermodynamic aspects. We determine the equilibrium phase diagram in temperature and C chemical potential, which allows us to predict the mesoscopic structure of the system that should be observed under experimentally relevant conditions.Comment: 5 pages, 3 figure

Finite element analysis and comparison of human skulls using MRI driven 3D printing

This is the author accepted manuscript. The final version is available via the link in this recordPoster abstrac

The Effect of iCook 4-H, a Childhood Obesity Prevention Program, on Blood Pressure and Quality of Life in Youth and Adults: A Randomized Control Trial

Objective: Obesity increases the risk of developing hypertension and from population-based samples with estimations that of 2-4% of the U.S. pediatric population has hypertension, which may affect quality of life. This study examined the effects of an obesity prevention program on blood pressure and quality of life in youth and adult participants. Methods: A multi-state research team recruited treatment dyads (youth and their adult meal preparer) to participate in a 12-week randomized control trial and follow-up through 24 months. The treatment group received a cooking and physical activity intervention, followed by booster sessions and mailed newsletters over the remaining two-year period. The control group received no intervention. Resting blood pressure and health related quality of life (HRQOL) surveys were administered at 0,4,12 and 24 months. Results: 228 dyads were recruited (n=77 control and n=151 for treatment). Youth and adult systolic blood pressure (SBP) increased over the 24 months (p=0.003 and p=0.03, respectively) with no differences between groups. From baseline to 24 months both control and treatment youths’ physical and psychological HRQOL increased (p=0.01 and p=0.002, respectively). At 0 and 4 months, youth and adult SBP was positively correlated (r=0.24, p=0.003 and r=0.33, p\u3c0.001, respectively). In the treatment group, there was an inverse association between adult SBP and youth psychological HRQOL at 4 months (r=-0.20, p=0.04), and a similar trend in adult SBP and youth physical HRQOL at 4 months in the treatment group (r=-0.19, p=0.05). Conclusion: A youth-adult dyad obesity prevention program consisting of culinary, mealtime and physical activity education, elicited improvements in HRQOL in youth participants