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Introduction: Proteases are enzymes that catalyze the hydrolysis of peptide bonds and play an important role in almost all biological processes. However, excessive protein proteolysis can be implicated in several diseases, such as cancer, as well as cardiovascular, inflammatory, neurodegenerative, bacterial, viral and parasitic diseases. In these cases, protease inhibitors can be used as one of versatile tools for regulating proteolytic activity of target proteases as well as therapeutic applications. In this study, we expressed and characterized a new serine protease inhibitory protein (PI-QT) from the metagenome of sponge-associated microorganisms in Escherichia coli. Methods: The gene PI-QT encoding for a new serine protease inhibitory protein was expressed in E. coli BL21(DE3). In addition, the expressed protein was purified and characterized. Results: Optimization of expression of the recombinant protein PI-QT in E. coli showed that suitable conditions for expression of the protein were pre-induction cell density (OD600) of 0.6 - 0.7, IPTG concentration of 1 mM and temperature of 25oC. The protease inhibitory protein was also purified and identified by mass spectrometry LC-MS/MS. The recombinant protein showed inhibitory activity against trypsin anda-chymotrypsin with activity values of 97526 U/mg and 41714 U/mg, respectively. Maximum activity of the protease inhibitory protein was obtained at pH 7 and temperature 20-35oC. The inhibitor was stable over pH 4-9 and up to temperature 50oC. Addition of Zn2+, Mg2+ and Ca2+ enhanced inhibitory activity, whereas other metal ions, surfactants and oxidants reduced inhibitory activity of the protease inhibitor. Conclusion: The recombinant protein PI-QT is a potential protease inhibitor for therapeutic applications.

This study aimed to assess the diversity and antimicrobial activity of cultivable bacteria associated with Vietnamese sponges. In total, 460 bacterial isolates were obtained from 18 marine sponges. Of these, 58.3% belonged to Proteobacteria, 16.5% to Actinobacteria, 18.0% to Firmicutes, and 7.2% to Bacteroidetes. At the genus level, isolated strains belonged to 55 genera, of which several genera, such as Bacillus, Pseudovibrio, Ruegeria, Vibrio, and Streptomyces, were the most predominant. Culture media influenced the cultivable bacterial composition, whereas, from different sponge species, similar cultivable bacteria were recovered. Interestingly, there was little overlap of bacterial composition associated with sponges when the taxa isolated were compared to cultivation-independent data. Subsequent antimicrobial assays showed that 90 isolated strains exhibited antimicrobial activity against at least one of seven indicator microorganisms. From the culture broth of the isolated strain with the strongest activity (Bacillus sp. M1_CRV_171), four secondary metabolites were isolated and identified, including cyclo(L-Pro-L-Tyr) (1), macrolactin A (2), macrolactin H (3), and 15,17-epoxy-16-hydroxy macrolactin A (4). Of these, compounds 2-4 exhibited antimicrobial activity against a broad spectrum of reference microorganisms.

INTRODUCTION AND AIMS: Gender and age patterns of drinking are important in guiding country responses to harmful use of alcohol. This study undertook cross-country analysis of drinking across gender, age groups in some high-and middle-income countries. DESIGN AND METHODS: Surveys of drinkers were conducted in Australia, England, Scotland, New Zealand, St Kitts and Nevis (high-income), Thailand, South Africa, Mongolia and Vietnam (middle-income) as part of the International Alcohol Control Study. Drinking pattern measures were high-frequency, heavier-typical quantity and higher-risk drinking. Differences in the drinking patterns across age and gender groups were calculated. Logistic regression models were applied including a measure of country-level income. RESULTS: Percentages of high-frequency, heavier-typical quantity and higher-risk drinking were greater among men than in women in all countries. Older age was associated with drinking more frequently but smaller typical quantities especially in high-income countries. Middle-income countries overall showed less frequent but heavier typical quantities; however, the lower frequencies meant the percentages of higher risk drinkers were lower overall compared with high-income countries (with the exception of South Africa). DISCUSSION AND CONCLUSIONS: High-frequency drinking was greater in high-income countries, particularly in older age groups. Middle-income countries overall showed less frequent drinking but heavier typical quantities. As alcohol use becomes more normalised as a result of the expansion of commercial alcohol it is likely frequency of drinking will increase with a likelihood of greater numbers drinking at higher risk levels.

Introduction and Aims To investigate behaviours related to four alcohol policy variables (policy-relevant behaviours) and demographic variables in relation to typical quantities of alcohol consumed on-premise in six International Alcohol Control study countries. Design and Methods General population surveys with drinkers using a comparable survey instrument and data analysed using path analysis in an overall model and for each country. Measures: typical quantities per occasion consumed on-premise; gender, age; years of education, prices paid, time of purchase, time to access alcohol and liking for alcohol advertisements. Results In the overall model younger people, males and those with fewer years of education consumed larger typical quantities. Overall lower prices paid, later time of purchase and liking for alcohol ads predicted consuming larger typical quantities; this was found in the high-income countries, less consistently in the high-middle-income countries and not in the low middle-income country. Three policy-relevant behaviours (prices paid, time of purchase, liking for alcohol ads) mediated the relationships between age, gender, education and consumption in high-income countries. Discussion and Conclusions International Alcohol Control survey data showed a relationship between policy-relevant behaviours and typical quantities consumed and support the likely effect of policy change (trading hours, price and restrictions on marketing) on heavier drinking. The path analysis also revealed policy-relevant behaviours were significant mediating variables between the effect of age, gender and educational status on consumption. However, this relationship is clearest in high-income countries. Further research is required to understand better how circumstances in low-middle-income countries impact effects of policies.

Introduction and Aims This study examines the proportion of alcohol markets consumed in harmful drinking occasions in a range of high-, middle-income countries and assesses the implications of these findings for conflict of interest between alcohol producers and public health and the appropriate role of the alcohol industry in alcohol policy space. Design and Methods Cross-sectional surveys were conducted in 10 countries as part of the International Alcohol Control study. Alcohol consumption was measured using location- and beverage-specific measures. A level of consumption defined as harmful use of alcohol was chosen and the proportion of the total market consumed in these drinking occasions was calculated for both commercial and informal alcohol. Results In all countries, sizeable proportions of the alcohol market were consumed during harmful drinking occasions. In general, a higher proportion of alcohol was consumed in harmful drinking occasions by respondents in the middle-income countries than respondents in the high-income countries. The proportion of informal alcohol consumed in harmful drinking occasions was lower than commercial alcohol. Discussion and Conclusions Informal alcohol is less likely to be consumed in harmful drinking occasions compared with commercial alcohol. The proportion of commercial alcohol consumed in harmful drinking occasions in a range of alcohol markets shows the reliance of the transnational alcohol corporations on harmful alcohol use. This reliance underpins industry lobbying against effective policy and support for ineffective approaches. The conflict of interest between the alcohol industry and public health requires their exclusion from the alcohol policy space.

Introduction and Aim: To report data on the implementation of alcohol policies regarding availability and marketing, and drink driving, along with ratings of enforcement from two small high-income to three high-middle income countries, and one low-middle income country. Method: This study uses the Alcohol Environment Protocol, an International Alcohol Control study research tool, which documents the alcohol policy environment by standardised collection of data from administrative sources, observational studies and interviews with key informants to allow for cross-country comparison and change over time. Results: All countries showed adoption to varying extents of key effective policy approaches outlined in the World Health Organization Global Strategy to Reduce the Harmful Use of Alcohol (2010). High-income countries were more likely to allocate resources to enforcement. However, where enforcement and implementation were high, policy on availability was fairly liberal. Key Informants judged alcohol to be very available in both high- and middle-income countries, reflecting liberal policy in the former and less implementation and enforcement and informal (unlicensed) sale of alcohol in the latter. Marketing was largely unrestricted in all countries and while drink-driving legislation was in place, it was less well enforced in middle-income countries. Conclusion: In countries with fewer resources, alcohol policies are less effective because of lack of implementation and enforcement and, in the case of marketing, lack of regulation. This has implications for the increase in consumption taking place as a result of the expanding distribution and marketing of commercial alcohol and consequent increases in alcohol-related harm. © 2018 The Authors Drug and Alcohol Review published by John Wiley & Sons Australia, Ltd on behalf of Australasian Professional Society on Alcohol and other Drugs

A 2010 World Health Assembly resolution called on member states to intensify efforts to address alcohol-related harm. Progress has been slow. This study aims to determine the magnitude of public support for 12 alcohol policies and whether it differs by country, demographic factors and drinking risk (volume consumed).

Introduction Taxation is increasingly being used as an effective means of influencing behaviour in relation to harmful products. In this paper we use data from six participating countries of the International Alcohol Control Study to examine and evaluate their comparative prices and tax regimes. Methods We calculate taxes and prices for three high-income and three middle-income countries. The data are drawn from the International Alcohol Control survey and from the Alcohol Environment Protocol. Tax systems are described and then the rates of tax on key products presented. Comparisons are made using the Purchasing Power Parity rates. The price and purchase data from each country's International Alcohol Control survey is then used to calculate the mean percentage of retail price paid in tax weighted by actual consumption. Results Both ad valorem and specific per unit of alcohol taxation systems are represented among the six countries. The prices differ widely between countries even though presented in terms of Purchasing Power Parity. The percentage of tax in the final price also varies widely but is much lower than the 75% set by the World Health Organization as a goal for tobacco tax. Conclusion There is considerable variation in tax systems and prices across countries. There is scope to increase taxation and this analysis provides comparable data, including the percentage of tax in final price, from some middle and high-income countries for consideration in policy discussion.

The alcohol industry have attempted to position themselves as collaborators in alcohol policy making as a way of influencing policies away from a focus on the drivers of the harmful use of alcohol (marketing, over availability and affordability). Their framings of alcohol consumption and harms allow them to argue for ineffective measures, largely targeting heavier consumers, and against population wide measures as the latter will affect moderate drinkers. The goal of their public relations organisations is to 'promote responsible drinking'. However, analysis of data collected in the International Alcohol Control study and used to estimate how much heavier drinking occasions contribute to the alcohol market in five different countries shows the alcohol industry's reliance on the harmful use of alcohol. In higher income countries heavier drinking occasions make up approximately 50% of sales and in middle income countries it is closer to two-thirds. It is this reliance on the harmful use of alcohol which underpins the conflicting interests between the transnational alcohol corporations and public health and which militates against their involvement in the alcohol policy arena. [Caswell S, Callinan S, Chaiyasong S, Cuong PV, Kazantseva E, Bayandorj T, Huckle T, Parker K, Railton R, Wall M. How the alcohol industry relies on harmful use of alcohol and works to protect its profits. Drug Alcohol Rev 2016;35:661-664].

Injuries result in nearly 6 million deaths and incur 52 million disability-adjusted life years annually, comprising 15% of the global disease burden. More than 90% of this burden occurs in low- and middle-income countries (LMICs). Given this burden, it’s not unexpected that injuries are the leading cause of death among travelers to LMICs, namely from road traffic crashes and drowning. Opportunely, the majority of injuries are preventable. Therefore, pre-travel advice regarding foreseeable dangers and how to avoid them may significantly mitigate injury risk, such as: wearing seatbelts, helmets and personal flotation devices when appropriate; responsibly consuming alcohol; and closely supervising children. Upon return, travelers to LMICs are in a unique position; having shared injury risks while abroad, travelers can advocate for injury control initiatives that might make the world safer for travelers and local populations alike.

Background Five countries in Asia including Bangladesh, China, India, Thailand and Vietnam formed a network called Asia-Pacific Network for Health Professional Education Reforms (ANHER). This network collectively conducted a survey at the national level and at the institutional level (for medical, nursing and public health education). We also undertook an assessment of final year graduates from these schools on their attitudes, competencies and willingness to work in rural areas. Methods Pretested anonymous questionnaire comprised of four sections including demographic data, attitudes towards working in rural area, where to work after graduation and perception about competency of respondents was used. Descriptive and analytical statistics were used for data analyses. Results About 60 % of students from Bangladesh and Thailand had positive attitude towards working in rural area, 50 % in both China and India and only 33 % in Vietnam. Students’ positive attitudes towards their school in terms of preparing or inspiring them to work in rural areas were low across all five countries. Upon graduation and in the next five years, majority of students wanted to work in public sectors. Interestingly confidence about overall competency was quite low. Discussion Positive attitude towards working in rural areas varied significantly across five countries in Asia. Medical schools should improve the preparation and inspiration towards working in rural areas for their students. Conclusion Medical schools should put more effort in improving students’ attitude towards working in rural areas. Electronic supplementary material The online version of this article (doi:10.1186/s12909-016-0719-9) contains supplementary material, which is available to authorized users.

Protease inhibitors (PIs) are proteins or peptides inhibiting the activity of protease and constitute a very important mechanism for regulating protease activity. So far, PIs have been used for the study of enzyme structures and reaction mechanisms, but recently they have been used in pharmaceutical, agricultural and industrial fields. Compared to terrestrial counterparts, marine environment possesses their unique characters, therefore, they are capable of producing a wide range of novel bioactive compounds including protease inhibitors. In our review, a brief overview of protease inhibitors (e.g., classification, mechanisms, and characteristics of PIs) and protease inhibitors from marine sponges and sponge-associated microorganisms has been reviewed.

Using culture - independent technique, hypervariable V4 region of 16S rDNA library sequencing by MiSEq, the baterial communities of three host sponges Rhabdastrella sp. DN, Spheciospongia sp. QT and Clathria sp. NT from Mien Trung sea were characterized. The phylogenetic analysis showed that bacterial community structures of the three investigated sponges similar to each other regarding 10 common phyla, although abundance of these phyla was different for each sponge. Phylum Thaumarchaeota was rich component for three sponges, especially in NT sponge (31.89%). In this sponge, 3 phyla Planctomycetes, Verrucomicrobia and Firmicutes were undetected in other 2 sponge samples. Phyla Cyanobacteria was observed only in DN sponge. The obtained amplicons were assigned in different taxonomic levels (class, order, family and genus) based on Silva database. At class level, Gammaproteobacteria was abundant in three sponges; and Caldilineae, Marine-group I were dominant in DN; mealwile, in QT other dominant classes were Marine-group I and Betaproteobacteria. For NT, they were Cytophaga and Deferribacteres. In general, all three sponges harbored abundant and genetically diverse microbial associated consortia and they shared several common bacterial operational taxonomic units, although with different abundance.

Foot and mouth disease virus (FMDV) causing infectious disease affects broadly cloven-hoofed animals. It has 7 different serotypes. However O is the most prevalent type in 3 founded types (O, A, Asia1) recurrence in Vietnam fields. The current vaccine for FMDV is inactivated or attenuated forms. Vaccines were able to raise strong immune responses but still caused the safety concerns. Virus-like particles could be a new vaccine generation that fulfills the present questions. On the one hand, it can tackle the safety issues, on the other hand, it can reserve FMDV intrinsic form which will provoke high immunogenicity. The important initial step to make virus-like particles is expression the capsid proteins of FMDV in appropriate system. So this study we describe how to design, express and purify of all capsid proteins: VP0, VP1 and VP3 of O type FMDV isolated in Vietnam field using SUMO fusion expression system.