Metallochaperones Are Needed for Mycobacterium tuberculosis and Escherichia coli Nicotinamidase-Pyrazinamidase Activity.

Mycobacterium tuberculosis nicotinamidase-pyrazinamidase (PZAse) is a metalloenzyme that catalyzes conversion of nicotinamide-pyrazinamide to nicotinic acid-pyrazinoic acid. This study investigated whether a metallochaperone is required for optimal PZAse activity. M. tuberculosis and Escherichia coli PZAses (PZAse-MT and PZAse-EC, respectively) were inactivated by metal depletion (giving PZAse-MT-Apo and PZAse-EC-Apo). Reactivation with the E. coli metallochaperone ZnuA or Rv2059 (the M. tuberculosis analog) was measured. This was repeated following proteolytic and thermal treatment of ZnuA and Rv2059. The CDC1551 M. tuberculosis reference strain had the Rv2059 coding gene knocked out, and PZA susceptibility and the pyrazinoic acid (POA) efflux rate were measured. ZnuA (200 μM) achieved 65% PZAse-EC-Apo reactivation. Rv2059 (1 μM) and ZnuA (1 μM) achieved 69% and 34.3% PZAse-MT-Apo reactivation, respectively. Proteolytic treatment of ZnuA and Rv2059 and application of three (but not one) thermal shocks to ZnuA significantly reduced the capacity to reactivate PZAse-MT-Apo. An M. tuberculosis Rv2059 knockout strain was Wayne positive and susceptible to PZA and did not have a significantly different POA efflux rate than the reference strain, although a trend toward a lower efflux rate was observed after knockout. The metallochaperone Rv2059 restored the activity of metal-depleted PZAse in vitro Although Rv2059 is important in vitro, it seems to have a smaller effect on PZA susceptibility in vivo. It may be important to mechanisms of action and resistance to pyrazinamide in M. tuberculosis Further studies are needed for confirmation.IMPORTANCE Tuberculosis is an infectious disease caused by the bacterium Mycobacterium tuberculosis and remains one of the major causes of disease and death worldwide. Pyrazinamide is a key drug used in the treatment of tuberculosis, yet its mechanism of action is not fully understood, and testing strains of M. tuberculosis for pyrazinamide resistance is not easy with the tools that are presently available. The significance of the present research is that a metallochaperone-like protein may be crucial to pyrazinamide's mechanisms of action and of resistance. This may support the development of improved tools to detect pyrazinamide resistance, which would have significant implications for the clinical management of patients with tuberculosis: drug regimens that are appropriately tailored to the resistance profile of a patient's individual strain lead to better clinical outcomes, reduced onward transmission of infection, and reduction of the development of resistant strains that are more challenging and expensive to treat

MODS-Wayne, a Colorimetric Adaptation of the Microscopic-Observation Drug Susceptibility (MODS) Assay for Detection of Mycobacterium tuberculosis Pyrazinamide Resistance from Sputum Samples.

Although pyrazinamide (PZA) is a key component of first- and second-line tuberculosis treatment regimens, there is no gold standard to determine PZA resistance. Approximately 50% of multidrug-resistant tuberculosis (MDR-TB) and over 90% of extensively drug-resistant tuberculosis (XDR-TB) strains are also PZA resistant. pncA sequencing is the endorsed test to evaluate PZA susceptibility. However, molecular methods have limitations for their wide application. In this study, we standardized and evaluated a new method, MODS-Wayne, to determine PZA resistance. MODS-Wayne is based on the detection of pyrazinoic acid, the hydrolysis product of PZA, directly in the supernatant of sputum cultures by detecting a color change following the addition of 10% ferrous ammonium sulfate. Using a PZA concentration of 800 µg/ml, sensitivity and specificity were evaluated at three different periods of incubation (reading 1, reading 2, and reading 3) using a composite reference standard (MGIT-PZA, pncA sequencing, and the classic Wayne test). MODS-Wayne was able to detect PZA resistance, with a sensitivity and specificity of 92.7% and 99.3%, respectively, at reading 3. MODS-Wayne had an agreement of 93.8% and a kappa index of 0.79 compared to the classic Wayne test, an agreement of 95.3% and kappa index of 0.86 compared to MGIT-PZA, and an agreement of 96.9% and kappa index of 0.90 compared to pncA sequencing. In conclusion, MODS-Wayne is a simple, fast, accurate, and inexpensive approach to detect PZA resistance, making this an attractive assay especially for low-resource countries, where TB is a major public health problem

Skilful prediction of Sahel summer rainfall on inter-annual and multi-year timescales

This is the final version of the article. Available from Springer Nature via the DOI in this record.Summer rainfall in the Sahel region of Africa exhibits one of the largest signals of climatic variability and with a population reliant on agricultural productivity, the Sahel is particularly vulnerable to major droughts such as occurred in the 1970s and 1980s. Rainfall levels have subsequently recovered, but future projections remain uncertain. Here we show that Sahel rainfall is skilfully predicted on inter-annual and multi-year (that is, >5 years) timescales and use these predictions to better understand the driving mechanisms. Moisture budget analysis indicates that on multi-year timescales, a warmer north Atlantic and Mediterranean enhance Sahel rainfall through increased meridional convergence of low-level, externally sourced moisture. In contrast, year-to-year rainfall levels are largely determined by the recycling rate of local moisture, regulated by planetary circulation patterns associated with the El Niño-Southern Oscillation. Our findings aid improved understanding and forecasting of Sahel drought, paramount for successful adaptation strategies in a changing climate.This work was supported by the Joint DECC/Defra Met Office Hadley Centre Climate Programme (GA01101) and the EU FP7 SPECS project. The contribution of D.P.R. has received funding from the NERC/DFID Future Climate for Africa programme under the AMMA-2050 project, grant number NE/M019977/1

Constitutively Activated PI3K Accelerates Tumor Initiation and Modifies Histopathology of Breast Cancer

The gene encoding phosphatidylinositol 3-kinase catalytic subunit α-isoform (PIK3CA, p110α) is frequently activated by mutation in human cancers. Based on detection in some breast cancer precursors, PIK3CA mutations have been proposed to have a role in tumor initiation. To investigate this hypothesis, we generated a novel mouse model with a Cre-recombinase regulated allele of p110α (myristoylated-p110α, myr-p110α) along with p53fl/fl deletion and KrasG12D also regulated by Cre-recombinase. After instillation of adenovirus-expressing Cre-recombinase into mammary ducts, we found that myr-p110α accelerated breast tumor initiation in a copy number-dependent manner. Breast tumors induced by p53fl/fl;KrasG12D with no or one copy of myr-p110α had predominantly sarcomatoid features, whereas two copies of myr-p110α resulted in tumors with a carcinoma phenotype. This novel model provides experimental support for importance of active p110α in breast tumor initiation, and shows that the amount of PI3K activity can affect the rate of tumor initiation and modify the histological phenotype of breast cancer

A programme for the prevention of post-traumatic stress disorder in midwifery (POPPY): indications of effectiveness from a feasibility study

Background: Midwives can experience events they perceive as traumatic when providingcare. As a result, some will develop post-traumatic stress disorder (PTSD), with adverse implications for their mental health, the quality of care provided for women and the employing organizations. POPPY (Programme for the prevention of PTSD in midwifery) is a package of educational and supportive resources comprising an educational workshop, information leaflet, peer support and access to trauma-focused clinical psychology intervention. A feasibility study of POPPY implementation was completed. Objective: This study aimed to identify potential impacts of POPPY on midwives’ understandingof trauma, their psychological well-being and job satisfaction. Method: POPPY was implemented in one hospital site. Before taking part in the POPPY workshop (T1) midwives (N = 153) completed self-report questionnaires, which measured exposure to work-related trauma, knowledge and confidence of managing trauma responses, professional impacts, symptoms of PTSD, burnout and job satisfaction. Measures were repeated (T2) approximately 6 months after training (n = 91, 62%). Results: Midwives’ confidence in recognizing (p = .001) and managing early traumaresponses in themselves and their colleagues significantly improved (both p < .001). There was a trend towards reduced levels of PTSD symptomatology, and fewer midwives reported sub clinical levels of PTSD (from 10% at T1 to 7% at T2). The proportion of midwives reporting high and moderate levels of depersonalization towards care was reduced (33% to 20%) and midwives reported significantly higher levels of job satisfaction at T2 (p < .001). Reductions in self-reported stress-related absenteeism (12% to 5%), long-term changes to clinical allocation (10% to 5%) and considerations about leaving midwifery (34% to 27%) were identified. Conclusions: In conclusion, POPPY shows very positive potential to improve midwives’ mental health and the sensitivity of care they provide, and reduce service disruption and costs for trusts. Large-scale longitudinal evaluation is required

Effect of Impurity Scattering on the Nonlinear Microwave Response in High-Tc Superconductors

We theoretically investigate intermodulation distortion in high-Tc superconductors. We study the effect of nonmagnetic impurities on the real and imaginary parts of nonlinear conductivity. The nonlinear conductivity is proportional to the inverse of temperature owing to the dependence of the damping effect on energy, which arises from the phase shift deviating from the unitary limit. It is shown that the final-states interaction makes the real part predominant over the imaginary part. These effects have not been included in previous theories based on the two-fluid model, enabling a consistent explanation for the experiments with the rf and dc fields

Innovation history of the CMAM Surge approach - Towards a shock-responsive health system in Kenya

This is the final version. Available from Oxford Policy Management via the link in this recordMaintains Working Paper 2Executive summary Given the increasing incidence of shocks, from droughts to COVID-19, and the weak capacity of most health systems to adequately respond to them, there is an urgent need to develop and scale up health system innovations that build shock-responsiveness. The Community-based Management of Acute Malnutrition (CMAM) Surge approach is one such innovation that aims to build the responsiveness of government health systems to seasonal and shock-related surges in demand for nutrition services in low-income contexts. Essentially, CMAM Surge involves health facilities setting thresholds (‘normal’, ‘alert’, ‘alarm’, and ‘emergency’) for acute malnutrition caseloads, based on a self-diagnosis of their capacity; monitoring trends in caseloads against these thresholds; and actioning internal health facility surge actions or seeking support from higher levels of government, the United Nations Children’s Fund (UNICEF), or non-governmental organisations (NGOs) when higher thresholds are crossed. This working paper advances the current knowledge on CMAM Surge (which is limited to country evaluations and perspective articles) by conducting an innovation history analysis of the approach, to understand the enablers of, and barriers to, its emergence and development; and its perceived potential (and limits) for building health system shock-responsiveness in Kenya. Developing this innovation history involved analysing the memories, reflections (through interviews), and documents of key implementing stakeholders in Kenya and internationally. Data were analysed using a widely applied social innovation conceptual framework that identifies six phases to a social innovation (Murray et al., 2010): diagnoses and prompts; proposals; piloting; scale-up; sustaining; and system change (towards a shock-responsive health system). The findings provide critical insights that can inform the scale-up of CMAM Surge to new contexts, improve the shock-responsiveness of CMAM Surge in Kenya, and guide the development of emerging adaptations to CMAM Surge, such as Health Surge, which seeks to adapt the surge approach principles and tools to build the shock-responsiveness of the entire health system.UK ai

Proteome Profiling of Breast Tumors by Gel Electrophoresis and Nanoscale Electrospray Ionization Mass Spectrometry

We have conducted proteome-wide analysis of fresh surgery specimens derived from breast cancer patients, using an approach that integrates size-based intact protein fractionation, nanoscale liquid separation of peptides, electrospray ion trap mass spectrometry, and bioinformatics. Through this approach, we have acquired a large amount of peptide fragmentation spectra from size-resolved fractions of the proteomes of several breast tumors, tissue peripheral to the tumor, and samples from patients undergoing noncancer surgery. Label-free quantitation was used to generate protein abundance maps for each proteome and perform comparative analyses. The mass spectrometry data revealed distinct qualitative and quantitative patterns distinguishing the tumors from healthy tissue as well as differences between metastatic and non-metastatic human breast cancers including many established and potential novel candidate protein biomarkers. Selected proteins were evaluated by Western blotting using tumors grouped according to histological grade, size, and receptor expression but differing in nodal status. Immunohistochemical analysis of a wide panel of breast tumors was conducted to assess expression in different types of breast cancers and the cellular distribution of the candidate proteins. These experiments provided further insights and an independent validation of the data obtained by mass spectrometry and revealed the potential of this approach for establishing multimodal markers for early metastasis, therapy outcomes, prognosis, and diagnosis in the future. © 2008 American Chemical Society

The detection of airborne transmission of tuberculosis from HIV-infected patients, using an in vivo air sampling model

Background. Nosocomial transmission of tuberculosis remains an important public health problem. We created an in vivo air sampling model to study airborne transmission of tuberculosis from patients coinfected with human immunodeficiency virus (HIV) and to evaluate environmental control measures. Methods. An animal facility was built above a mechanically ventilated HIV‐tuberculosis ward in Lima, Peru. A mean of 92 guinea pigs were continuously exposed to all ward exhaust air for 16 months. Animals had tuberculin skin tests performed at monthly intervals, and those with positive reactions were removed for autopsy and culture for tuberculosis. Results. Over 505 consecutive days, there were 118 ward admissions by 97 patients with pulmonary tuberculosis, with a median duration of hospitalization of 11 days. All patients were infected with HIV and constituted a heterogeneous group with both new and existing diagnoses of tuberculosis. There was a wide variation in monthly rates of guinea pigs developing positive tuberculin test results (0%–53%). Of 292 animals exposed to ward air, 159 developed positive tuberculin skin test results, of which 129 had laboratory confirmation of tuberculosis. The HIV‐positive patients with pulmonary tuberculosis produced a mean of 8.2 infectious quanta per hour, compared with 1.25 for HIV‐negative patients with tuberculosis in similar studies from the 1950s. The mean monthly patient infectiousness varied greatly, from production of 0–44 infectious quanta per hour, as did the theoretical risk for a health care worker to acquire tuberculosis by breathing ward air. Conclusions. HIV‐positive patients with tuberculosis varied greatly in their infectiousness, and some were highly infectious. Use of environmental control strategies for nosocomial tuberculosis is therefore a priority, especially in areas with a high prevalence of both tuberculosis and HIV infection