Na+/K+ carrier ionophore antibiotics valinomycin and monensin enhance the antibacterial activity of fluoride.

Fluoride is routinely used as a highly effective antibacterial agent that interferes with bacterial metabolism through fundamentally different mechanisms. One of the major bacterial evasion mechanisms against fluoride is the impermeability of cell envelope to the anion that limits its cellular uptake. Therefore, translating such compounds to clinical settings requires novel mechanisms to facilitate the uptake of membrane-impermeant molecules. Published data have indicated antibiotic synergy between fluoride and membrane destabilizing agents that induce strong fluoride toxicity in bacteria via enhancing the permeability of bacterial membranes to fluoride. Here, we report a similar mechanism of antibiotic synergy between fluoride and potassium ion carriers, valinomycin and monensin against Gram-positive bacteria, B. subtilis and S. aureus. Molecular dynamics simulations were performed to understand the effect of potassium on the binding affinity of fluoride to monensin and valinomycin. The trajectory results strongly indicated that the monensin molecules transport fluoride ions across the cell membrane via formation of ion-pair between the monensin-K+ complex and a fluoride. This study provides new insights to design novel compounds to enhance the uptake of small toxic anions via synergistic interactions and thus exert strong antibacterial activity against a wide variety of pathogens.

Involvement of alcohol in injury cases in rural Sri Lanka: prevalence and associated factors among in-patients in three primary care hospitals.

BACKGROUND: Injuries account for a major proportion of global morbidity and mortality related to alcohol use. Information on the prevalence of alcohol-related injury in rural Sri Lanka is limited. The aims of this study were to determine the burden of alcohol-related injury in a hospital-based sample in rural Sri Lanka and explore factors associated with an increased risk of alcohol-related injury. METHODS: Involvement of alcohol in injury amongst in-patients was assessed in three hospitals in the North Central Province of Sri Lanka over 6 months. Adult (≥ 18 years) patients were eligible. Patients were assessed for: injury characteristics, current alcohol use (in the past year) using the Alcohol Use Disorder Identification Test (AUDIT), and acute intoxication. Patients with a blood alcohol concentration (BAC) reading equivalent of 10 mg/dL (2.17 mmol/L) were considered as having an alcohol-related injury. Binary logistic regression was used to explore association between alcohol-related injury and demographic and injury characteristics. RESULTS: A total of 883 injured patients were eligible and consented to the study. No alcohol use was reported by 487 (55.2%) of patients (35.6% of men, 95.2% of women). Prevalence of alcohol-related injuries was 14.8% overall and 32.8% among current alcohol users. Almost all patients with an alcohol-related injury were male (122/123; 99.2%); 24 (18.8%) of these patients scored positive for possible alcohol dependence. Patients with an alcohol-related injury had significantly higher AUDIT scores (median = 15 vs 6, p < 0.001), were significantly more likely to be aged 26-40 (OR 2.29, 95% CI:1.11, 4.72) or 41-55 years (OR 2.76, 95% CI: 1.29, 5.90) (compared to 18-25 years), to have a transport-related injury (OR 5.14, 95% CI: 2.30, 11.49) (compared to animal/plant sting/bite), and have intentional injuries (OR 3.47, 95% CI: 1.01, 11.87). CONCLUSIONS: One in three injuries among people who drank alcohol in this sample were alcohol-related. In addition, problematic alcohol use was higher among those with alcohol-related injury. Further work is needed to explore whether this prevalence of alcohol-related injury is reflected in other rural settings in Sri Lanka.

Assessment of current status and geospatial analysis of compliance to bacteriological parameters: Tools to ensure access to safe water in Sri Lanka.

Compliance of drinking-water to bacteriological parameters serves as a surrogate measure of the risk of water-borne diseases. Understanding the risk of water-borne diseases could help promote healthy behaviors such as household water treatment and safe water storage practices and advocacy to increase access to centrally-managed piped water. The objective of this research was to assess the current status of compliance and to geospatially analyze the probability of compliance to bacteriological parameters in the Western Province of Sri Lanka. A drinking-water quality survey was conducted among 4508 households representing four water-source types: National Water Supply and Drainage Board (NWSDB), dug wells, Community Water Supply (CWS), and tube wells, and other sources. Besides, a detailed assessment of selected NWSDB and CWS supplies was done. Water samples were tested for the total coliform count, thermotolerant coliform count, and free residual chlorine levels against the Sri Lanka Standards. Indicator kriging was performed using the geospatial analyst tool of ArcGIS version 10.6 for different water source types to interpolate the probability of compliance for both total coliform count and thermotolerant coliform count. The bacteriological compliance decreased from NWSDB to tube wells and other sources to CWS to dug wells. The interpolation maps confirm the relatively higher compliance of NWSDB for bacteriological parameters compared to other sources. Areas with a high probability of compliance for both parameters show considerable overlap with urban areas with a supply of centrally managed water from the NWSDB. It is recommended to expand the coverage of NWSDB water, strengthen the drinking-water quality surveillance system and water safety plans, and promote household water treatment and safe storage practices in the Western Province of Sri Lanka.

Socioeconomic position and suicidal behaviour in rural Sri Lanka: a prospective cohort study of 168,000+ people.

PURPOSE: Lower socioeconomic position (SEP) is associated with an increased risk of suicidal behaviour in high income countries, but this association is not established in low- and middle-income countries (LMIC). METHODS: We investigated the association of SEP with suicidal behaviour in a prospective cohort study of 168,771 Sri Lankans followed up for episodes of attempted suicide and suicide. SEP data were collected at baseline at the household and individual level at the start of the follow-up period. We used multilevel Poisson regression models to investigate the association of SEP at community, household and individual levels with attempted suicide/suicide. RESULTS: Lower levels of asset ownership [IRR (95% CI) suicide 1.74 (0.92, 3.28); attempted suicide 1.67 (1.40, 2.00)] and education [suicide 3.16 (1.06, 9.45); attempted suicide 2.51 (1.70, 3.72)] were associated with an increased risk of suicidal behaviour. The association of these measures of SEP and attempted suicide was stronger in men than women. Individuals living in deprived areas [1.42 (1.16, 1.73)] and in households with a young female head of household [1.41 (1.04, 1.93)] or a temporary foreign migrant [1.47 (1.28, 1.68)] had an elevated risk of attempted suicide. Farmers and daily wage labourers had nearly a doubling in risk of attempted suicide compared to other occupations. CONCLUSIONS: Improved employment opportunities, welfare and mental health support services, as well as problem-solving skills development, may help support individuals with poorer education, farmers, daily wage labourers, individuals in young female-headed households and temporary foreign migrant households.

Attempted suicide in Sri Lanka - An epidemiological study of household and community factors.

BACKGROUND: An individual's suicide risk is determined by personal characteristics, but is also influenced by their environment. Previous studies indicate a role of contextual effects on suicidal behaviour, but there is a dearth of quantitative evidence from Asia. METHODS: Individual and community level data were collected on 165,233 people from 47,919 households in 171 communities in rural Sri Lanka. Data were collected on individual (age, sex, past suicide attempts and individual socioeconomic position (SEP)) and household (household SEP, pesticide access, alcohol use and multigenerational households) level factors. We used 3-level logit models to investigate compositional (individual) and contextual (household/community) effects. RESULTS: We found significant variation between households 21% (95% CI 18%, 24%) and communities 4% (95% CI 3%, 5%) in the risk of a suicide attempt. Contextual factors as measured by low household SEP (OR 2.37 95% CI 2.10, 2.67), low community SEP (OR 1.45 95% CI 1.21, 1.74), and community 'problem' alcohol use (OR 1.44 95% CI 1.19, 1.75) were associated with an increased risk of suicide attempt. Women living in households with alcohol misuse were at higher risk of attempted suicide. We observed a protective effect of living in multigenerational households (OR 0.53 95% CI 0.42, 0.65). LIMITATIONS: The outcome was respondent-reported and refers to lifetime reports of attempted suicide, therefore this study might be affected by socially desirable responding. CONCLUSIONS: Our study finds that contextual factors are associated with an individual's risk of attempted suicide in Sri Lanka, independent of an individual's personal characteristics.

Is socioeconomic position associated with risk of attempted suicide in rural Sri Lanka? A cross-sectional study of 165†000 individuals.

BACKGROUND: Lower socioeconomic position (SEP) is associated with an increased risk of suicidal behaviour in high-income countries, but this association is unclear in low-income and middle-income countries. METHODS: We investigated the association of SEP with attempted suicide in a cross-sectional survey of 165 233 Sri Lankans. SEP data were collected at the household (assets, social standing (highest occupation of a household member), foreign employment and young (≤40 years) female-headed households) and individual level (education and occupation). Respondent-reported data on suicide attempts in the past year were recorded. Random-effects logistic regression models, accounting for clustering, were used to investigate the association of SEP with attempted suicide. RESULTS: Households reported 398 attempted suicides in the preceding year (239 per 100 000). Fewer assets (OR 3.2, 95% CI 2.4 to 4.4) and having a daily wage labourer (ie, insecure/low-income job; OR 2.3, 95% CI 1.6 to 3.2) as the highest occupation increased the risk of an attempted suicide within households. At an individual level, daily wage labourers were at an increased risk of attempted suicide compared with farmers. The strongest associations were with low levels of education (OR 4.6, 95% CI 2.5 to 8.4), with a stronger association in men than women. CONCLUSIONS: We found that indicators of lower SEP are associated with increased risk of attempted suicide in rural Sri Lanka. Longitudinal studies with objective measures of suicide attempts are needed to confirm this association. TRIAL REGISTRATION NUMBER: NCT01146496; Pre-results.

Determination of maternal pedigree and ewe-lamb spatial relationships by application of Bluetooth technology in extensive farming systems.

The objectives of this study were to validate the application of Bluetooth technology to determine maternal pedigree and to determine ewe-lamb spatial relationships in extensive farming systems. A total of 35 first-cross Merino ewes (Merino × Border Leicester and East Friesian) and 23 of their lambs aged 1 to 3 wk were fitted with activity monitors equipped with Bluetooth (BT) technology (ActiGraph wGT3X-BT) by means of halters and collars, respectively. The BT devices on lambs were programmed to receive wireless signals once every minute from nearby BT units on ewes, which were programmed as beacons sending BT signals 4 times every second. Ewes and lambs fitted with sensors were dispatched into the paddocks, and after 10 d, the sensor units were retrieved and the BT signals received by lambs were downloaded using the ActiGraph software. The maternal pedigree of the lambs was determined as the ewe from which the lamb received the most BT signals. The distance between the lamb receiving the signal and the ewe sending the signal was estimated from the strength of BT signal received. The pedigree determined by BT was compared with the pedigree determined by DNA profiling and verification. The results showed that the accuracy of maternal pedigree determined by BT signals reached 100% within the first 15 min of returning animals to pasture of ewes and lambs fitted with sensors. Maternal signals (counts/d) received by 1-, 2-, and 3-wk-old lambs were 617 ± 102, 603 ± 54, and 498 ± 36, respectively, and the corresponding nonmaternal signals received were 140 ± 27, 106 ± 30, and 155 ± 39, respectively. Maternal signals received during the dark period were significantly higher than the maternal signals received during the light period ( < 0.05). Maternal signals received during the light period by 3-wk-old lambs were significantly lower when compared with those received by 1- and 2-wk-old lambs. Over 90% of the BT signals received from within 2 m of the lamb were from its mother. The maternal BT signals expressed as a portion of total BT signals decreased with increasing distance from the lamb. The results show that BT wireless networking is a fast and reliable method for the determination of maternal pedigree of lambs in extensive farming systems. In addition, wireless BT technology is also useful in determining mother-offspring spatial relationships.

A supplement containing trans-10, cis-12 conjugated linoleic acid reduces milk fat yield but does not alter organ weight or body fat deposition in lactating ewes.

Conjugated linoleic acids (CLA) have been demonstrated to be a potent inhibitor of milk fat synthesis in ruminants, but effects on carcass composition and organ weight are unknown. Our objectives in this experiment were to determine the dose response of ruminally protected CLA on the performance, organ weight, and fatty acid (FA) composition of early lactation dairy ewes. Twenty-four multiparous dairy ewes were fed a basal diet for 10 wk that was supplemented with a lipid-encapsulated CLA at 1 of 3 levels: no CLA (control, CON), low CLA (L-CLA), or high CLA (H-CLA) to supply 0, 1.5, or 3.8 g/d, respectively, of both trans-10, cis-12 and cis-9, trans-11 CLA. Dry matter intake was not affected (P > 0.05) by dietary treatment. Ewes fed H-CLA had a 13% higher milk yield compared with those receiving either CON or L-CLA. Compared with CON, milk fat yield (g/d) was 14 and 24% lower in ewes fed L-CLA or H-CLA, respectively. Supplementing ewes with CLA did not affect carcass or organ weights, carcass composition, or organ FA content. Compared with ewes receiving the CON diet, CLA supplementation had little effect on the FA composition of the Longissimus dorsi, although cis-9, trans-11 and trans-10, cis-12 CLA were increased in ewes receiving H-CLA. The current findings are consistent with the view that the energy spared by the CLA reduction in milk fat content was mainly partitioned to milk yield and there was no evidence of organ hypertrophy or liver steatosis.

The role of private drug vendors as malaria treatment providers in selected malaria endemic areas of Sri Lanka.

BACKGROUND AND OBJECTIVES: The involvement of private drug vendors in malaria treatment is particularly high in developing countries and understanding their practices and knowledge about antimalarials and malaria treatment will aid in devising strategies to increase the correct use of antimalarials and improve adherence to the government's malaria drug policy. Results of a study on the knowledge and practices of the private drug vendors conducted in seven districts in Sri Lanka, mostly in malarious areas are presented. METHODS: Data on awareness of government's malaria drug policy, practice of issuing antimalarials, knowledge about malaria and antimalarial drugs were collected from the drug vendors using pre-tested questionnaire in vernacular language. Data were statistically analysed using Stata 8.2. Chi-square test was carried out for individual explanatory variables and a logistic regression model was applied taking all response variables as binary outcome. RESULTS: Vendors' knowledge on antimalarials was poor with 58% of the vendors being unaware of the government malaria drug policy in the country. Also, the advice provided to customers buying antimalarials was limited. However, the majority of the private vendors emphasised that they were aware of the importance of case confirmation before treatment as stressed in the national policy. Although, the vendors did not have a high awareness of national drug policies they were only found selling chloroquine and primaquine as recommended by the Ministry of Health. INTERPRETATION AND CONCLUSION: In recent years Sri Lanka, as a whole, has experienced very little malaria. The reduction in demand for antimalarials due to low incidence levels may have influenced the knowledge and awareness on antimalarials and government drug policies. However, since low levels of malaria do not guarantee that epidemics will not occur, attempts to educate private drug vendors as a part of an organised control programmes are of major importance.

A rapid and efficient system for screening HIV-1 Pol mRNA-specific ribozymes.

Hammerhead ribozymes are potentially important tools for suppressing intracellular expression of unwanted RNAs. However, the reports that exist on their activity against different targets have described mixed success. As an initial step towards developing a rapid and effective system for in vivo testing of ribozymes, two human immunodeficiency virus type-1 (HIV-1) polymerase (Pol) mRNA-specific ribozymes, RzPro directed against the protease (Pro) coding region and RzRT directed against the reverse transcriptase (RT) coding region, were designed and tested in Escherichia coli. Both ribozymes displayed similar efficiencies in cleaving their target RNAs in vitro. RNA polymerase chain reaction was adapted to demonstrate the in vivo cleavage of RzPro and RzRT target sites. The resultant drop in HIV-1 RT activity was measured as well. The degree of suppression of RT activity was more apparent in vivo in cells expressing RzRT. The RT activity in cells expressing RzRT was shown to decrease by up to 96%. This system will be useful for rapid screening of (i) other ribozyme target sites within the Pol mRNA so that multitargeted ribozymes could be designed for use in anti-HIV-1 gene therapy, (ii) ribozymes with improved stability and catalytic activity, and (iii) cofactors, if any that could enhance ribozyme activity in vivo.

Resistance to human immunodeficiency virus type 1 (HIV-1) infection in human CD4+ lymphocyte-derived cell lines conferred by using retroviral vectors expressing an HIV-1 RNA-specific ribozyme.

Toward gene therapy for the treatment of human immunodeficiency virus type 1 (HIV-1) infections in AIDS, Moloney murine leukemia virus-derived retroviral vectors were engineered to allow constitutive and tat-inducible expression of an HIV-1 5' leader sequence-specific ribozyme (Rz1). These vectors were used to infect the human CD4+ lymphocyte-derived MT4 cell line. The stable MT4 transformants expressing an HIV-1 RNA-specific ribozyme, under the control of the herpes simplex virus thymidine kinase (tk) promoter, were found to be somewhat resistant to HIV-1 infection as virus production was delayed. In cells allowing ribozyme expression under control of the simian virus 40 or cytomegalovirus promoter, the rate of HIV-1 multiplication was slightly decreased, and virus production was delayed by about 14 days. The highest level of resistance to HIV-1 infection was observed in MT4 cells transformed with a vector containing a fusion tk-TAR (trans activation-responsive) promoter to allow ribozyme expression in a constitutive and tat-inducible manner; no HIV-1 production was observed 22 days after infection of these cells. These results indicate that retroviral vectors expressing HIV-1 RNA-specific ribozymes can be used to confer resistance to HIV-1 infection.