World Health Organization Priority Research Articles

Antibiotic resistance prediction for Mycobacterium tuberculosis from genome sequence data with Mykrobe.

Two billion people are infected with Mycobacterium tuberculosis, leading to 10 million new cases of active tuberculosis and 1.5 million deaths annually. Universal access to drug susceptibility testing (DST) has become a World Health Organization priority. We previously developed a software tool, Mykrobe predictor, which provided offline species identification and drug resistance predictions for M. tuberculosis from whole genome sequencing (WGS) data. Performance was insufficient to support the use of WGS as an alternative to conventional phenotype-based DST, due to mutation catalogue limitations. Here we present a new tool, Mykrobe, which provides the same functionality based on a new software implementation. Improvements include i) an updated mutation catalogue giving greater sensitivity to detect pyrazinamide resistance, ii) support for user-defined resistance catalogues, iii) improved identification of non-tuberculous mycobacterial species, and iv) an updated statistical model for Oxford Nanopore Technologies sequencing data. Mykrobe is released under MIT license at https://github.com/mykrobe-tools/mykrobe. We incorporate mutation catalogues from the CRyPTIC consortium et al. (2018) and from Walker et al. (2015), and make improvements based on performance on an initial set of 3206 and an independent set of 5845 M. tuberculosis Illumina sequences. To give estimates of error rates, we use a prospectively collected dataset of 4362 M. tuberculosis isolates. Using culture based DST as the reference, we estimate Mykrobe to be 100%, 95%, 82%, 99% sensitive and 99%, 100%, 99%, 99% specific for rifampicin, isoniazid, pyrazinamide and ethambutol resistance prediction respectively. We benchmark against four other tools on 10207 (=5845+4362) samples, and also show that Mykrobe gives concordant results with nanopore data. We measure the ability of Mykrobe-based DST to guide personalized therapeutic regimen design in the context of complex drug susceptibility profiles, showing 94% concordance of implied regimen with that driven by phenotypic DST, higher than all other benchmarked tools.

IJU this issue

In this issue, seven Original Articles with four Editorial Comments, two Review Articles and three Urological Notes were published in the International Journal of Urology. Urinary tract infection or perioperative infection control is one of the important clinical issues for urologists. A rapid increase of carbapenem-resistant Enterobacteriaceae and its outbreak is a grave worldwide problem. Kitano et al. (Hiroshima, Japan) reviewed mechanisms behind antimicrobial resistance, and new antimicrobials to treat resistant microbes. This article concisely describes the current state of resistant microbes associated with urinary tract infections and countermeasures for antimicrobial resistance, including new antimicrobials. As the author denotes in Table 3 of that article, the World Health Organization priority list for antibiotic-resistant bacteria should be recognized by urologists. As different treatment modalities for localized prostate cancer yield similar patient outcomes, the focus in determining appropriate treatment has shifted toward health-related quality of life. Murakami et al. (Kanagawa, Japan) reviewed the impact of brachytherapy and other modalities on health-related quality of life. Except for the inferior outcome regarding urinary obstruction and irritation, brachytherapy is likely to yield similar or better health-related quality of life than other treatment modalities, such as operation, active surveillance or external beam radiation therapy. Due to scant data on prospective direct comparison among modalities and recent replacement by sophisticated technologies, such as intensity-modulated radiation therapy or robot-assisted radical prostatectomy, it seems hardly difficult to draw a convincing conclusion. Sazuka et al. (Chiba, Japan) retrospectively examined the impact of post-void residual urine volume on the risk of postoperative intravesical recurrence in 81 patients with upper urinary tract urothelial carcinoma undergoing nephroureterectomy. The authors found the presence of pyuria and a residual urine volume (defined as ≥30 mL) were associated with bladder recurrence in the multivariate analysis, and the 2-year postoperative recurrence-free rates of patients with and without a residual urine volume were 32% and 82%, respectively. Although the number of the cohort for a propensity-score match study is limited, the authors’ hypothesis is unique and is worthwhile for a large-scale prospective analysis. Choo et al. (Suwon, Korea) carried out to a web-based questionnaire survey regarding the individual treatment trend and adherence to international guidelines on non-muscle-invasive bladder cancer. Out of 2334 urologists who were active members of the Japanese Society of Urological Oncology, the Korean Urological Oncology Society and the Taiwan Urological Association, 701 (30%) responded to the survey from June 2016 to February 2017. The authors found significant differences in the management of intermediate- to high-risk non-muscle-invasive bladder cancer between Japan, Korea and Taiwan. As the authors described, East Asian countries are geographically close together, and share ethnic and sociocultural similarities, such that the survey can serve as the basis for joint efforts among East Asian urologists to develop common clinical guidelines for the management of non-muscle-invasive bladder cancer. Nephrostomy tube after percutaneous intrarenal surgeries, such as percutaneous nephrolithotomy (PCNL), sometimes prevents a short hospital stay and adversely affects patients’ quality of life. A tubeless procedure would be ideal after PCNL. Takeda et al. (Miyazaki, Japan) compared the efficacy and safety among patients who received trans-tract electrocoagulation at the end of endoscopic combined intrarenal surgery (225 patients) with that of those who did not (72 patients). Although stone number or size was in favor of the trans-tract electrocoagulation group, the nephrostomy tube-free rate and other postoperative clinical parameters were significantly higher in the trans-tract electrocoagulation group. As Kakinoki (Saga, Japan) indicated in the Editorial Comment, all patients do not always require such procedure, particularly when miniaturized PCNL is used. To clarify the necessity of hemostasis, a prospective randomized study comparing miniaturized PCNL is warranted. None declared.

Genetically Modified Rabies Virus Vector-Based Rift Valley Fever Virus Vaccine is Safe and Induces Efficacious Immune Responses in Mice.

Rift Valley fever virus (RVFV), which causes Rift Valley fever (RVF), is a mosquito-borne zoonotic pathogen that causes serious morbidity and mortality in livestock and humans. RVF is a World Health Organization (WHO) priority disease and, together with rabies, is a major health burden in Africa. Here, we present the development and characterization of an inactivated recombinant RVFV and rabies virus (RABV) vaccine candidate (rSRV9-eGn). Immunization with rSRV9-eGn stimulated the production of RVFV-specific IgG antibodies and induced humoral and cellular immunity in mice but did not induce the production of neutralizing antibodies. IgG1 and IgG2a were the main isotypes observed by IgG subtype detection, and IgG3 antibodies were not detected. The ratios of IgG1/IgG2a > 1 indicated a Type 2 humoral immune response. An effective vaccine is intended to establish a long-lived population of memory T cells, and mice generated memory cells among the proliferating T cell population after immunization with rSRV9-eGn, with effector memory T cells (TEM) as the major population. Due to the lack of prophylactic treatment experiments, it is impossible to predict whether this vaccine can protect animals from RVFV infection with only high titres of anti-RVFV IgG antibodies and no neutralizing antibodies induced, and thus, protection confirmation needs further verification. However, this RVFV vaccine designed with RABV as the vector provides ideas for the development of vaccines that prevent RVFV and RABV infections.

Treating drug-resistant wound pathogens with non-medicated dressings: an in vitro study.

To assess the in vitro antimicrobial performance of a non-medicated hydro-responsive wound dressing (HRWD) on the sequestration and killing of wound relevant microorganisms found on the World Health Organization (WHO) priority pathogens list. Suspensions of Pseudomonas aeruginosa, Acinetobacter baumannii and methicillin-resistant Staphylococcus aureus (MRSA) were placed on petri dishes. Dressings were each placed on top, incubated for 30 minutes and then removed from the inoculated petri dish. The surface of the dressings previously in contact with the bacterial suspensions were placed directly onto a tryptone soy agar (TSA) plate and incubated for 24 hours. Dressings were then removed from the TSA plate and the level of bacterial growth on the plates was assessed. Sequestered microorganism viability was assessed using LIVE/DEAD viability kits and visualisation by epifluorescence. Our results indicated that HRWDs sequester and retain Pseudomonas aeruginosa, Acinetobacter baumannii and MRSA within the dressing. Non-medicated HRWDs containing bound PHMB (polyhexamethylene biguanide, HRWD+PHMB) killed the microorganisms sequestered within the dressing matrix. These data suggest that non-medicated HRWD+PHMB is an effective against WHO priority pathogens and promoting goal of antimicrobial stewardship in wound care.

Modeling Antibacterial Activity with Machine Learning and Fusion of Chemical Structure Information with Microorganism Metabolic Networks.

Predicting the activity of new chemical compounds over pathogenic microorganisms with different metabolic reaction networks (MRN s) is an important goal due to the different susceptibility to antibiotics. The ChEMBL database contains >160 000 outcomes of preclinical assays of antimicrobial activity for 55 931 compounds with >365 parameters of activity (MIC, IC50, etc.) and >90 bacteria strains of >25 bacterial species. In addition, the Leong and Barabàsi data set includes >40 MRNs of microorganisms. However, there are no models able to predict antibacterial activity for multiple assays considering both drug and MRN structures at the same time. In this work, we combined perturbation theory, machine learning, and information fusion techniques to develop the first PTMLIF model. The best linear model found presented values of specificity = 90.31/90.40 and sensitivity = 88.14/88.07 in training/validation series. We carried out a comparison to nonlinear artificial neural network (ANN) techniques and previous models from the literature. Next, we illustrated the practical use of the model with an experimental case of study. We reported for the first time the isolation and characterization of terpenes from the plant Cissus incisa. The antibacterial activity of the terpenes was experimentally determined. The more active compounds were phytol and α-amyrin, with MIC = 100 μg/mL for Vancomycin-resistant Enterococcus faecium and Acinetobacter baumannii resistant to carbapenems. These compounds are already known from other sources. However, they have been isolated and evaluated for the first time here against several strains of multidrug-resistant bacteria including World Health Organization (WHO) priority pathogens. Last, we used the model to predict the activity of these compounds versus other microorganisms with different MRNs in order to find other potential targets.

1,4,7-Triazacyclononane Restores the Activity of β-Lactam Antibiotics against Metallo-β-Lactamase-Producing Enterobacteriaceae: Exploration of Potential Metallo-β-Lactamase Inhibitors.

Metallo-β-lactamase (MBL)-producing Enterobacteriaceae are of grave clinical concern, particularly as there are no metallo-β-lactamase inhibitors approved for clinical use. The discovery and development of MBL inhibitors to restore the efficacy of available β-lactams are thus imperative. We investigated a zinc-chelating moiety, 1,4,7-triazacyclononane (TACN), for its inhibitory activity against clinical carbapenem-resistant Enterobacteriaceae MICs, minimum bactericidal concentrations (MBCs), the serum effect, fractional inhibitory concentration indexes, and time-kill kinetics were determined using broth microdilution techniques according to Clinical and Laboratory Standards Institute (CSLI) guidelines. Enzyme kinetic parameters and the cytotoxic effects of TACN were determined using spectrophotometric assays. The interactions of the enzyme-TACN complex were investigated by computational studies. Meropenem regained its activity against carbapenemase-producing Enterobacteriaceae, with the MIC decreasing from between 8 and 64 mg/liter to 0.03 mg/liter in the presence of TACN. The TACN-meropenem combination showed bactericidal effects with an MBC/MIC ratio of ≤4, and synergistic activity was observed. Human serum effects on the MICs were insignificant, and TACN was found to be noncytotoxic at concentrations above the MIC values. Computational studies predicted that TACN inhibits MBLs by targeting their catalytic active-site pockets. This was supported by its inhibition constant (Ki ), which was 0.044 μM, and its inactivation constant (Kinact), which was 0.0406 min-1, demonstrating that TACN inhibits MBLs efficiently and holds promise as a potential inhibitor.IMPORTANCE Carbapenem-resistant Enterobacteriaceae (CRE)-mediated infections remain a significant public health concern and have been reported to be critical in the World Health Organization's priority pathogens list for the research and development of new antibiotics. CRE produce enzymes, such as metallo-β-lactamases (MBLs), which inactivate β-lactam antibiotics. Combination therapies involving a β-lactam antibiotic and a β-lactamase inhibitor remain a major treatment option for infections caused by β-lactamase-producing organisms. Currently, no MBL inhibitor-β-lactam combination therapy is clinically available for MBL-positive bacterial infections. Hence, developing efficient molecules capable of inhibiting these enzymes could be a promising way to overcome this phenomenon. TACN played a significant role in the inhibitory activity of the tested molecules against CREs by potentiating the activity of carbapenem. This study demonstrates that TACN inhibits MBLs efficiently and holds promises as a potential MBL inhibitor to help curb the global health threat posed by MBL-producing CREs.

A Low-Cost, Community Knowledge Approach to Estimate Maternal and Jaundice-Associated Mortality in Rural Bangladesh.

In the absence of a civil registration system, a house-to-house survey is often used to estimate cause-specific mortality in low- and middle-income countries. However, house-to-house surveys are resource and time intensive. We applied a low-cost community knowledge approach to identify maternal deaths from any cause and jaundice-associated deaths among persons aged ≥ 14 years, and stillbirths and neonatal deaths in mothers with jaundice during pregnancy in five rural communities in Bangladesh. We estimated the method's sensitivity and cost savings compared with a house-to-house survey. In the five communities with a total of 125,570 population, we identified 13 maternal deaths, 60 deaths among persons aged ≥ 14 years associated with jaundice, five neonatal deaths, and four stillbirths born to a mother with jaundice during pregnancy over the 3-year period before the survey using the community knowledge approach. The sensitivity of community knowledge method in identifying target deaths ranged from 80% for neonatal deaths to 100% for stillbirths and maternal deaths. The community knowledge approach required 36% of the staff time to undertake compared with the house-to-house survey. The community knowledge approach was less expensive but highly sensitive in identifying maternal and jaundice-associated mortality, as well as all-cause adult mortality in rural settings in Bangladesh. This method can be applied in rural settings of other low- and middle-income countries and, in conjunction with hospital-based hepatitis diagnoses, used to monitor the impact of programs to reduce the burden of cause-specific hepatitis mortality, a current World Health Organization priority.

Influenza Vaccinations for All Pregnant Women? Better Evidence Is Needed

Pregnant women are a World Health Organization (WHO) priority group for influenza vaccination, but evidence of effectiveness and safety for pregnant women comes from observational studies, which are notoriously prone to confounding by indication and healthy-vaccinee bias. The latter type of bias leads to an overestimation of the effectiveness and safety of the vaccine, which may be what occurs in pregnant women. Indeed, better educated women with healthier behaviors and who seek better medical care may be more adherent to vaccinations recommended by doctors, scientific societies and health authorities. Therefore, it is fundamental to obtain information about vaccine effectiveness and safety from randomized controlled trials (RCTs). Cochrane reviews have identified only one RCT with “low risk of bias”. Its results were unclear in terms of maternal, perinatal, and infant deaths and hospitalization, and showed a Number Needed to Vaccine (NNV) of 55 for mothers, with an excess of local adverse effects. A Cochrane review concluded that the inactivated influenza vaccine provides pregnant women with uncertain or very limited protection against influenza-like illnesses and influenza. Some observational studies have suggested possible adverse effects of the inflammation following the vaccination. Consistent with the Cochrane reviewers’ conclusions, further trials for influenza vaccines with appropriate study designs and comparison groups are required before promoting universal seasonal influenza vaccinations of pregnant women. Meanwhile, vaccination in second to third trimester should be offered while communicating the uncertainties that still exist, promoting informed choices. Vaccination in the first trimester is debatable and debated. This does not mean leaving women defenseless; many other useful behavioral and environmental measures can reduce infectious disease.

A mapping of nursing and midwifery research in the Eastern Mediterranean Region, 2006-2016.

Nurses and midwives constitute the largest group of health care professionals globally. Challenges to these professions make it difficult to set regional priorities for policies and research development. The purpose of this study was to map current nursing and midwifery research in the Eastern Mediterranean Region. Nursing and midwifery schools were identified by each country's nursing and midwifery board and ministries of education/public health. Information was collected for the years 2006-2016 via surveys, websites, Google scholar and expert informants. A total of 299 schools were identified and 241 of these were contacted; 85 completed surveys from 15 countries were analysed. A total of 1116 research topics covered by 3287 publications were identified, many of which were clustered into the five World Health Organization priority areas. The least developed areas were disaster management and emergency preparedness. This study provides a database of nursing and midwifery research in the Region. Some gaps were identified based on the research priorities of the Region, but these gaps could be addressed by close collaboration among local researchers.

Rose Bengal-Mediated Photoinactivation of Multidrug Resistant Pseudomonas aeruginosa Is Enhanced in the Presence of Antimicrobial Peptides.

Due to the overuse of antibiotics in medicine and food production, and their targeted mechanism of action, an increasing rate in spreading of antibiotic resistance genes has been noticed. This results in inefficient therapy outcomes and higher mortality all over the world. Pseudomonas aeruginosa (carbapenem-resistant) is considered one of the top three critical species according to the World Health Organization’s priority pathogens list. This means that new drugs and/or treatments are needed to tackle infections caused by this bacterium. In this context search for new/alternative approaches that would overcome resistance to classical antimicrobials is of prime importance. The use of antimicrobial photodynamic inactivation (aPDI) and antimicrobial peptides (AMPs) is an efficient strategy to treat localized infections caused by multidrug-resistant P. aeruginosa. In this study, we have treated P. aeruginosa cells photodynamically in the presence and in the absence of AMP (CAMEL or pexiganan). The conditions for aPDI were as follows: rose bengal (RB) as a photosensitizing agent at 1–10 μM concentration, and subsequent irradiation with 514 nm-LED at 23 mW/cm2 irradiance. The analysis of cell number after the treatment has shown that the combined action of RB-mediated aPDI and cationic AMPs reduced the number of viable cells below the limit of detection (<1log10 CFU/ml). This was in contrast to no reduction or partial reduction after aPDI or AMP applied separately. Students t-test was applied to test the statistical significance of the results. Noteworthy, our treatment proved to be effective against all 35 clinical isolates of P. aeruginosa tested within this study, including those characterized as multiresistant. Moreover, we demonstrated that such treatment is safe and does not violate the growth dynamics of human keratinocytes (77.3–97.64% survival depending on the concentration of the studied compounds or their mixtures).

Global Health

Global Health

Priorities of positive mental health promotion in the Iranian community: a qualitative study.

BackgroundPositive Mental Health (PMH) enables people to cope with the common stresses of life and adversity to achieve their full potential and humanity. In many communities, promoting PMH via prioritized interventions has been considered as a key component of public health policies to optimize mental well-being.ObjectiveTo set the priorities of interventional programs of Iranian PMH promotion according to the World Health Organization (WHO) Priority Public Health Condition (PPHC) analytical framework.MethodsThis qualitative study was implemented in 2017 in Tehran, Iran and had two main phases. In a qualitative needs-assessment phase, needs of the community’s PMH were collected through eight focus group discussions with a general population aged 30 to 60 years-old. In a priority-setting phase, the priorities of PMH were extracted through an expert panel consisting of mental health professionals and policy makers. Data gathering was implemented based on purposeful sampling according to inclusion criteria. Data were analyzed based on directional content analysis using Dedoose software version 7.6.6.ResultsFifty-one people and ten mental health professionals and policymakers participated in this study. The process of data analyzing, categorized PMH needs in 4 main categories, 15 subcategories and 46 codes. The four categories were financial security, social security, healthy lifestyle and promoting psychological factors. In the expert panel, first, the indicators of PMH priority setting were determined and then based on the priority public health condition analytical framework of the World Health Organization, the most important of PMH priority in each level was indicated as “creating job positions” in socioeconomic level, “providing proper working conditions” in differential exposure, “promoting practical life skills training” in the differential vulnerability, “easy and affordable access to mental health services” in differential healthcare outcome.ConclusionAppropriate policymaking and regulation at national level regarding employment, promoting working conditions, and reducing unemployment, promote community PMH as well as expanding accessible and affordable mental health services in the national healthcare system and empowering the community through providing practical life skill courses.

Emergence and spread of carbapenem-resistant Acinetobacter baumannii international clones II and III in Lima, Peru

Carbapenem-resistant Acinetobacter baumannii is the top-ranked pathogen in the World Health Organization priority list of antibiotic-resistant bacteria. It emerged as a global pathogen due to the successful expansion of a few epidemic lineages, or international clones (ICs), producing acquired class D carbapenemases (OXA-type). During the past decade, however, reports regarding IC-I isolates in Latin America are scarce and are non-existent for IC-II and IC-III isolates. This study evaluates the molecular mechanisms of carbapenem resistance and the epidemiology of 80 non-duplicate clinical samples of A. baumannii collected from February 2014 through April 2016 at two tertiary care hospitals in Lima. Almost all isolates were carbapenem-resistant (97.5%), and susceptibility only remained high for colistin (95%). Pulsed-field gel electrophoresis showed two main clusters spread between both hospitals: cluster D containing 51 isolates (63.8%) associated with sequence type 2 (ST2) and carrying OXA-72, and cluster F containing 13 isolates (16.3%) associated with ST79 and also carrying OXA-72. ST2 and ST79 were endemic in at least one of the hospitals. ST1 and ST3 OXA-23-producing isolates were also identified. They accounted for sporadic hospital isolates. Interestingly, two isolates carried the novel OXA-253 variant of OXA-143 together with an upstream novel insertion sequence (ISAba47). While the predominant A. baumannii lineages in Latin America are linked to ST79, ST25, ST15, and ST1 producing OXA-23 enzymes, we report the emergence of highly resistant ST2 (IC-II) isolates in Peru producing OXA-72 and the first identification of ST3 isolates (IC-III) in Latin America, both considered a serious threat to public health worldwide.

THE RELATIONSHIP BETWEEN SUICIDE IDEATION, DEPRESSIVE SYMPTOMS AND PERSONALITY TRAITS IN ELDERLY: PILOT STUDY RESULTS

Healthy aging and the research on the related factors is one of the World Health Organization's priorities for the 2020-2030 years. Depression and suicidal ideation can adversely affect an individual's aging experience. Personality traits are important factors that allow to predict the individual's behavior in different situations, as well as the accurance of emotional difficulties. The aim of the study was to investigate the relationship between symptoms of depression, suicidal ideation and personality traits in elderly, and to determine whether the relationship between personality traits and suicidal ideation remains statistically significant after controlling the severity of depression symptoms. Thirty nine respondents aged 65 and over were included in the pilot study (M = 71.23; SD = 4.95). The Latvian Clinical Personality Inventory (LCPI) was used for data collection. The results indicate a statistically significant relationship between several personality traits, depression symptoms and suicidal ideation for elderly. Controlling the severity of depression symptoms, statistically significant correlations were found between suicidal ideation and personality traits such as distrust, social withdrawal, self-harm, dissociation proneness on facet level and introversion on domain level. The results of the pilot study largely coincide with the results of previous studies and indicate that certain personality traits, irrespective of the degree of severity of depression symptoms, can explain an additional variation in suicidal thoughts and, hence, the risk of suicide for elderly. It would be important to conduct a study with similar variables in a larger sample.

Tobacco control achievements and priority areas in the WHO Europe Region: A review.

INTRODUCTIONTobacco control efforts have been advancing globally, including the adoption and entry into force of the World Health Organization (WHO) Framework Convention of Tobacco Control (FCTC), as well as the adoption of the European Union EU Tobacco Products Directive. With the present review, the European Network for Smoking and Tobacco Prevention (ENSP) and European Respiratory Society (ERS) aim to provide a comprehensive overview on the status of WHO FCTC implementation, policy achievements and priority areas across countries in the WHO Europe Region.METHODSThe review was conducted through a triangulation of data extracted from a survey administered to ENSP members, the WHO FCTC Implementation Database, Tobacco Control Laws and the Tobacco Control Scale 2016.RESULTSUsing the WHO MPOWER measures as a framework, we report on the implementation status of nine FCTC articles across 47 countries in the WHO Europe Region. The average number of articles fully implemented was 3.58. FCTC articles least fully implemented were: Article 5.3 on Tobacco Industry Interference (25.5%, n=12), Article 20 on Research (34.0%, n=16), and Article 15 on Illicit trade (40.4%, n=19). The most commonly fully implemented articles were: Article 8 on Smoke-free legislation (63.8%, n=30), Article 16 on Underage sales (57.4%, n=27), and Article 6 on Price and tax measures (51.2%, n=24). Policy achievements and recommended priority areas for future national tobacco control activities varied greatly among countries.CONCLUSIONSFindings highlight the multitude of successes in tobacco control efforts across the region in recent years, but point out the need to address gaps in FCTC implementation.

Using an Assessment Tool to Further Tobacco Control Accomplishment in Thailand.

Background:Because implementation of the Framework Convention on Tobacco Control (FCTC), a World Health Organization (WHO) treaty to reduce tobacco use, is an important goal of the 2015 United Nations Sustainable Development Goals (SDGs) and Thailand has sought to fully comply with all its articles, a multiperspective assessment was developed to ensure that any gaps in compliance with FCTC provisions were identified and addressed.Method:One assessment mechanism of this multicomponent assessment was the development by experts and use by stakeholders of a 3-tiered rating of all major provisions of the main articles of the FCTC. The results of the performance ratings on FCTC articles by a diverse group of stakeholders were used to spotlight areas of local and regional implementation and compliance with FCTC provisions.Results:Implementation ratings by stakeholders generally followed the chronology of WHO priorities as reflected in the development by WHO of guidelines for the various FCTC articles with highest ratings for articles 6, 8, 11, 12, 14, 15, and 16. However, only 5 articles (Articles 6, 11, 12, 15, and 16) reached level 2 (effectiveness) of the 3-level rating; articles 6, 8, 11, 13, and 14 are discussed because they are the WHO priority articles of the MPOWER tobacco control policy. Importantly, stakeholders cited problems with lack of completeness of present Thai law and processes for enforcement, as well as lack of public understanding regarding tobacco control strategies and provisions.Conclusions:Overall, the breadth and inclusiveness of the stakeholder approach devised for improving implementation by the Tobacco Control Research and Knowledge Management Center provided greater understanding about shortcomings of present policy and resource management which informed the Tobacco Products Control Act passed in 2017 and plans for advancing stronger Thai regulation by local and national government.

Design and Solution-Phase Synthesis of Membrane-Targeting Lipopeptides with Selective Antibacterial Activity.

Designing selective antibacterial molecules remains an unmet goal in the field of membrane-targeting agents. Herein, we report the rational design and synthesis of a new class of lipopeptides, which possess highly selective bacterial killing over mammalian cells. The selective interaction with bacterial over mammalian membranes was established through various spectroscopic, as well as microscopic experiments, including biophysical studies with the model membranes. A detailed antibacterial structure-activity relationship was delineated after preparing a series of molecules consisting of the peptide moieties with varied sequence of amino acids, such as d-phenylalanine, d-leucine, and d-lysine. Antibacterial activity was found to vary with the nature and positioning of hydrophobicity in the molecules, as well as number of positive charges. Optimized lipopeptide 9 did not show any hemolytic activity even at 1000 μg mL-1 and displayed >200-fold and >100-fold selectivity towards S. aureus and E. coli, respectively. More importantly, compound 9 was found to display good antibacterial activity (MIC 6.3-12.5 μg mL-1 ) against the five top most critical bacteria according to World Health Organization (WHO) priority pathogens list. Therefore, the results suggested that this new class of lipopeptides bear real promises for the development as future antibacterial agents.

Mycobacterium ulcerans low infectious dose and mechanical transmission support insect bites and puncturing injuries in the spread of Buruli ulcer.

Addressing the transmission enigma of the neglected disease Buruli ulcer (BU) is a World Health Organization priority. In Australia, we have observed an association between mosquitoes harboring the causative agent, Mycobacterium ulcerans, and BU. Here we tested a contaminated skin model of BU transmission by dipping the tails from healthy mice in cultures of the causative agent, Mycobacterium ulcerans. Tails were exposed to mosquito (Aedes notoscriptus and Aedes aegypti) blood feeding or punctured with sterile needles. Two of 12 of mice with M. ulcerans contaminated tails exposed to feeding A. notoscriptus mosquitoes developed BU. There were no mice exposed to A. aegypti that developed BU. Eighty-eight percent of mice (21/24) subjected to contaminated tail needle puncture developed BU. Mouse tails coated only in bacteria did not develop disease. A median incubation time of 12 weeks, consistent with data from human infections, was noted. We then specifically tested the M. ulcerans infectious dose-50 (ID50) in this contaminated skin surface infection model with needle puncture and observed an ID50 of 2.6 colony-forming units. We have uncovered a biologically plausible mechanical transmission mode of BU via natural or anthropogenic skin punctures.

Status of vaccine research and development for enterotoxigenic Escherichia coli

Enterotoxigenic Escherichia coli (ETEC) is one of the most common bacterial causes of diarrhea-associated morbidity and mortality, particularly among infants and young children in developing countries. Still, the true impact on child and traveler health is likely underestimated. There are currently no licensed vaccines for ETEC, but studies indicate high public health impact, cost-effectiveness, and feasibility of immune protection through vaccination. ETEC vaccine development remains a World Health Organization priority. Traditionally, ETEC vaccine development efforts have focused on inducing antitoxin and anticolonization antigen immunity, as studies indicate that antibodies against both antigen types can contribute to protection and thus have potential for vaccines. Leading cellular vaccine candidates are ETVAX (a mixture of four inactivated strains) and ACE527 (a mixture of three live attenuated strains), both of which have been found to be safe and immunogenic in Phase 1/2 trials. ETVAX is the furthest along in development with descending-age studies already underway in Bangladesh. Other ETEC vaccine candidates based on protein subunits, toxoids (both LT and ST), or novel, more broadly conserved ETEC antigens are also under development. Of these, a protein adhesin-based subunit approach is the most advanced. Impact and economic models suggest favorable vaccine cost-effectiveness, which may help expand market interest in ETEC vaccines. Combination vaccine formulations may help improve the economic case for development and use, and better point-of-care diagnostics will help to raise awareness of the true health burden of ETEC and highlight the potential public health benefit of ETEC vaccine introduction. Better diagnostics and vaccine demand forecasting will also improve vaccine development financing and support accelerated uptake once a licensed vaccine becomes available.

Secretion of dengue virus envelope protein ectodomain from mammalian cells is dependent on domain II serotype and affects the immune response upon DNA vaccination.

Dengue virus (DENV) is currently among the most important human pathogens and affects millions of people throughout the tropical and subtropical regions of the world. Although it has been a World Health Organization priority for several years, there is still no efficient vaccine available to prevent infection. The envelope glycoprotein (E), exposed on the surface on infective viral particles, is the main target of neutralizing antibodies. For this reason it has been used as the antigen of choice for vaccine development efforts. Here we show a detailed analysis of factors involved in the expression, secretion and folding of E ectodomain from all four DENV serotypes in mammalian cells, and how this affects their ability to induce neutralizing antibody responses in DNA-vaccinated mice. Proper folding of E domain II (DII) is essential for efficient E ectodomain secretion, with DIII playing a significant role in stabilizing soluble dimers. We also show that the level of protein secreted from transfected cells determines the strength and efficiency of antibody responses in the context of DNA vaccination and should be considered a pivotal feature for the development of E-based DNA vaccines against DENV.