In vitro antimicrobial activity of natural toxins and animal venoms tested against Burkholderia pseudomallei

BACKGROUND: Burkholderia pseudomallei are the causative agent of melioidosis. Increasing resistance of the disease to antibiotics is a severe problem in treatment regime and has led to intensification of the search for new drugs. Antimicrobial peptides are the most ubiquitous in nature as part of the innate immune system and host defense mechanism. METHODS: Here, we investigated a group of venoms (snakes, scorpions and honey bee venoms) for antimicrobial properties against two strains of Gram-negative bacteria Burkholderia pseudomallei by using disc-diffusion assay for in vitro susceptibility testing. The antibacterial activities of the venoms were compared with that of the isolated L-amino acid oxidase (LAAO) and phospholipase A(2 )(PLA(2)s) enzymes. MICs were determined using broth dilution method. Bacterial growth was assessed by measurement of optical density at the lowest dilutions (MIC 0.25 mg/ml). The cell viability was measured using tetrazolium salts (XTT) based cytotoxic assay. RESULTS: The studied venoms showed high antimicrobial activity. The venoms of C. adamanteus, Daboia russelli russelli, A. halys, P. australis, B. candidus and P. guttata were equally as effective as Chloramphenicol and Ceftazidime (30 μg/disc). Among those tested, phospholipase A(2 )enzymes (crotoxin B and daboiatoxin) showed the most potent antibacterial activity against Gram-negative (TES) bacteria. Naturally occurring venom peptides and phospholipase A(2 )proved to possess highly potent antimicrobial activity against Burkholderia pseudomallei. The XTT-assay results showed that the cell survival decreased with increasing concentrations (0.05–10 mg/mL) of Crotalus adamanteus venom, with no effect on the cell viability evident at 0.5 mg/mL. CONCLUSION: This antibacterial profile of snake venoms reported herein will be useful in the search for potential antibacterial agents against drug resistant microorganisms like B. pseudomallei

Adaptive Evolution of the Venom-Targeted vWF Protein in Opossums that Eat Pitvipers

The rapid evolution of venom toxin genes is often explained as the result of a biochemical arms race between venomous animals and their prey. However, it is not clear that an arms race analogy is appropriate in this context because there is no published evidence for rapid evolution in genes that might confer toxin resistance among routinely envenomed species. Here we report such evidence from an unusual predator-prey relationship between opossums (Marsupialia: Didelphidae) and pitvipers (Serpentes: Crotalinae). In particular, we found high ratios of replacement to silent substitutions in the gene encoding von Willebrand Factor (vWF), a venom-targeted hemostatic blood protein, in a clade of opossums known to eat pitvipers and to be resistant to their hemorrhagic venom. Observed amino-acid substitutions in venom-resistant opossums include changes in net charge and hydrophobicity that are hypothesized to weaken the bond between vWF and one of its toxic snake-venom ligands, the C-type lectin-like protein botrocetin. Our results provide the first example of rapid adaptive evolution in any venom-targeted molecule, and they support the notion that an evolutionary arms race might be driving the rapid evolution of snake venoms. However, in the arms race implied by our results, venomous snakes are prey, and their venom has a correspondingly defensive function in addition to its usual trophic role

Protocol for a sequential, prospective meta-analysis to describe coronavirus disease 2019 (COVID-19) in the pregnancy and postpartum periods.

We urgently need answers to basic epidemiological questions regarding SARS-CoV-2 infection in pregnant and postpartum women and its effect on their newborns. While many national registries, health facilities, and research groups are collecting relevant data, we need a collaborative and methodologically rigorous approach to better combine these data and address knowledge gaps, especially those related to rare outcomes. We propose that using a sequential, prospective meta-analysis (PMA) is the best approach to generate data for policy- and practice-oriented guidelines. As the pandemic evolves, additional studies identified retrospectively by the steering committee or through living systematic reviews will be invited to participate in this PMA. Investigators can contribute to the PMA by either submitting individual patient data or running standardized code to generate aggregate data estimates. For the primary analysis, we will pool data using two-stage meta-analysis methods. The meta-analyses will be updated as additional data accrue in each contributing study and as additional studies meet study-specific time or data accrual thresholds for sharing. At the time of publication, investigators of 25 studies, including more than 76,000 pregnancies, in 41 countries had agreed to share data for this analysis. Among the included studies, 12 have a contemporaneous comparison group of pregnancies without COVID-19, and four studies include a comparison group of non-pregnant women of reproductive age with COVID-19. Protocols and updates will be maintained publicly. Results will be shared with key stakeholders, including the World Health Organization (WHO) Maternal, Newborn, Child, and Adolescent Health (MNCAH) Research Working Group. Data contributors will share results with local stakeholders. Scientific publications will be published in open-access journals on an ongoing basis

Community engagement for malaria elimination in contested areas of the Karen/Kayin State, Myanmar: A case study on the Malaria Elimination Task Force

Background This study evinces how the community engagement (CE) approach of the Malaria Elimination Task Force (METF) in the Karen/Kayin State of southeast Myanmar contributed to generating participation in program activities, such as early detection and treatment, malaria blood surveys and mass drug administration. Methods We investigated the CE approach through ethnographic and key-informant unstructured interviews as well as focus group discussions and participant observation with program participants METF team members. Interview transcripts were analysed manually applying inductive content analysis. Results In the context of the Thai-Myanmar border, CE had two main dimensions: The first was to enable the politically sensitive entry of the program into intervention areas, the second was to create mutual understanding among the METF team and people from the population and gain trust. For the first, the METF needed to acknowledge, get permission and communicate with a complex landscape of governing actors and leaders. For the second, creating mutual understanding meant to spend time learning about villagers’ lifeworlds and adapting engagement accordingly to create understanding of the program in resonance with these lifeworlds. Gaining trust meant to leverage leadership and build trust through informal “horizontal” engagement and acts of caring. Three key dimensions of villagers’ lifeworlds that appeared most crucial for staff to learn about were: villagers’ experience with the outside world; leadership and governing relationships to villagers: authority and distrust; and their perceived need for malaria care. Conclusions Successful CE is highly context specific. CE success was based on the adaptability of the approach towards the complex political and socio-cultural contexts of the different areas of the Karen/Kayin State. Hence, CE is not a fixed approach or strategy defined before entering the setting, but constantly evolving based on the encounters with stakeholders and people of the population.</p

Bank Credit Receipt, Executors of the Estate of J. H. Woodward, First National Bank, Birmingham, Alabama, June 19, 1918

A document from an extensive collection spanning four generations of the Woodward family that operated merchant pig iron companies in West Virginia and Alabama. The collection begins with Stimpson Harvey Woodward (S. H. Woodward), a native of Massachusetts, who moved from Pittsburgh to Wheeling, West Virginia in 1852. He had interests in an iron company as early as 1852 in West Virginia and began Alabama operations in 1869. The family business continued in Alabama until the death of S. H. Woodward's great-grandson in 1965

Community engagement for malaria elimination in contested areas of the Karen/Kayin State, Myanmar: A case study on the Malaria Elimination Task Force

Background This study evinces how the community engagement (CE) approach of the Malaria Elimination Task Force (METF) in the Karen/Kayin State of southeast Myanmar contributed to generating participation in program activities, such as early detection and treatment, malaria blood surveys and mass drug administration. Methods We investigated the CE approach through ethnographic and key-informant unstructured interviews as well as focus group discussions and participant observation with program participants METF team members. Interview transcripts were analysed manually applying inductive content analysis. Results In the context of the Thai-Myanmar border, CE had two main dimensions: The first was to enable the politically sensitive entry of the program into intervention areas, the second was to create mutual understanding among the METF team and people from the population and gain trust. For the first, the METF needed to acknowledge, get permission and communicate with a complex landscape of governing actors and leaders. For the second, creating mutual understanding meant to spend time learning about villagers’ lifeworlds and adapting engagement accordingly to create understanding of the program in resonance with these lifeworlds. Gaining trust meant to leverage leadership and build trust through informal “horizontal” engagement and acts of caring. Three key dimensions of villagers’ lifeworlds that appeared most crucial for staff to learn about were: villagers’ experience with the outside world; leadership and governing relationships to villagers: authority and distrust; and their perceived need for malaria care. Conclusions Successful CE is highly context specific. CE success was based on the adaptability of the approach towards the complex political and socio-cultural contexts of the different areas of the Karen/Kayin State. Hence, CE is not a fixed approach or strategy defined before entering the setting, but constantly evolving based on the encounters with stakeholders and people of the population.</p

Low seroprevalence of HIV and syphilis in pregnant women in refugee camps on the Thai-Burma border.

SUMMARY: Refugee and migrant populations are considered to be at high risk from sexually transmitted infection (STI) and HIV. Cross-sectional surveys for syphilis and HIV were conducted in antenatal clinics (ANCs) on the Thai-Burmese border. In Mae La refugee camp, the seroprevalence of HIV and syphilis were 0.2% (one of 500) (95% CI 0-1.1) and 0% (0 of 404) (95% CI 0-0.9) in 1997; and 0.4% (two of 500) (95% CI 0.1-1.4) and 0.4% (three of 741) (95% CI 0.1-1.2) in 2005, respectively; syphilis seroprevalence in migrant women in 2005 was 0 (0 of 234) (95% CI 0-1.6). The seroprevalence was lower than that reported from surrounding ANCs for Thai or Burmese women. Focus group discussions with HIV-negative and -positive pregnant refugee women established that aspects of Karen culture and isolation (geographical and political) had a significant protective role from HIV and STI. This survey has resulted in programmatic changes in services to pregnant women in this area

Accelerated training of skilled birth attendants in a marginalized population on the Thai-Myanmar border: A multiple methods program evaluation

Background To evaluate a skilled birth attendant (SBA) training program in a neglected population on the Thai-Myanmar border, we used multiple methods to show that refugee and migrant health workers can be given effective training in their own environment to become SBAs and teachers of SBAs. The loss of SBAs through resettlement to third countries necessitated urgent training of available workers to meet local needs. Methods and Findings All results were obtained from student records of theory grades and clinical log books. Qualitative evaluation of both the SBA and teacher programs was obtained using semi-structured interviews with supervisors and teachers. We also reviewed perinatal indicators over an eight-year period, starting prior to the first training program until after the graduation of the fourth cohort of SBAs. Results Four SBA training programs scheduled between 2009 and 2015 resulted in 79/88 (90%) of students successfully completing a training program of 250 theory hours and 625 supervised clinical hours. All 79 students were able to: achieve pass grades on theory examination (median 80%, range [70–89]); obtain the required clinical experience within twelve months; achieve clinical competence to provide safe care during childbirth. In 2010–2011, five experienced SBAs completed a train-the-trainer (TOT) program and went on to facilitate further training programs. Perinatal indicators within Shoklo Malaria Research Unit (SMRU), such as place of birth, maternal and newborn outcomes, showed no significant differences before and after introduction of training or following graduate deployment in the local maternity units. Confidence, competence and teamwork emerged from qualitative evaluation by senior SBAs working with and supervising students in the clinics. Conclusions We demonstrate that in resource-limited settings or in marginalized populations, it is possible to accelerate training of skilled birth attendants to provide safe maternity care. Education needs to be tailored to local needs to ensure evidence-based care of women and their families.</p