Efficient Cloning of Inserts for Phage Display by Golden Gate Assembly.

Phage display enables the discovery of high-affinity binders. In phage display, one commonly uses traditional cloning methods to insert DNA into the coding region of one of the five capsid proteins. Here we describe the use of a new vector with kanamycin resistance and BsaI sites for the utilization of Golden Gate cloning into the N-terminus of mature protein III. We also describe the successful pentavalent display of six different inserts: the AviD-tag, the Z-domain of protein A, the Myc-tag, the ALFA nanobody, the BC2 nanobody, and the Flag-tag.

Introduction of two novel species of Hymenopellis (Agaricales, Physalacriaceae) from Thailand.

Hymenopellis is the most diverse genus in the group of oudemansielloid/xeruloid taxa (Physalacriaceae). This genus has a worldwide distribution with records mostly from Europe and America. Asian taxa are least represented. In this paper on Hymenopellis from Thailand, two novel species are introduced, and a Hymenopellis collection affine to H.orientalis is described. Macro and micromorphological characters are described. Maximum likelihood and Bayesian phylogenetic analyses were performed on combined ITS and nrLSU regions to confirm taxonomical placement and infer the phylogenetic affinities of the studied species. Hymenopellisstraminea sp. nov. is straw-yellow, with medium-sized basidiomata, abundant and diverse in form cheilocystidia, few, narrowly lageniform to fusiform pleurocystidia, and clamp connections at the lower part of the stipe. Hymenopellisutriformis sp. nov. has mostly utriform pleurocystidia and 2-spored basidia. In the inferred phylogenies, the new species from this study formed distinct clades well supported by bootstrap proportions and posterior probabilities. The studied specimen affine to H.orientalis produced 2-spored basidia whereas published descriptions of other specimens mention 4-spored basidia. Moreover, the genetic distance between ITS sequences of this specimen and that of a Hymenopellisorientalis specimen from GenBank was 1.30-2.57%. Therefore, the conspecificity of our specimen with H.orientalis is uncertain, and additional specimens are needed to fully confirm its identity.

Facilitated hospital-to-pre-hospital feedback for professional development (PHEM Feedback): a service evaluation using a self-reported questionnaire to understand the experiences of participating pre-hospital clinicians in the first year of operation.

Background: Information governance and resource challenges can impede pre-hospital clinicians from accessing and reflecting upon clinical information from the hospital phase of care, to ascertain how appropriate their diagnoses and management were. The authors performed a 12-month service evaluation of a hospital-to-pre-hospital feedback system, in which clinical information was requested by pre-hospital clinicians, and returned by a small team of hospital-based clinicians, while meeting information governance standards. Method: Pre-hospital clinicians in one ambulance station and one air ambulance service accessed patient information from a hospital, via a mediating senior pre-hospital colleague (a facilitator). Case-based learning conversations between the facilitator and clinician followed, using a report from the hospital. Evidence of benefit to the pre-hospital clinicians was prospectively collected using Likert-type scales, regarding general satisfaction, likelihood to change practice and effects on well-being. Reports aimed to be generated by the hospital within 14 days. Results: All 59 appropriate requests had reports returned. Of the reports, 59.5% were returned in 14 days or less. The median duration was 11 days (interquartile range 7-25). Learning conversations were completed in 86.4% (n = 51) of these cases, and of those, clinician questionnaires were completed in 66.7% (n = 34). Of the 34 questionnaire respondents, 82.4% (n = 28) were very satisfied with the returned information. A total of 61.1% (n = 21) were either likely or very likely to change their practice following the hospital's information, and 64.7% (n = 22) reported similar or very similar impressions to the hospital's eventual diagnosis. Regarding mental health, 76.5% (n = 26) reported positively or very positively affected mental health, while 2.9% (n = 1) reported adversely affected mental health. All of the respondents, 100% (n = 34) were either satisfied or very satisfied with the learning conversation. Conclusion: While hospital-based clinical information was successfully and securely provided to pre-hospital clinicians, these pilot data suggest it is not possible to meet the self-imposed, empirical 14-day target with four to five voluntary doctors. Sustained performance may improve with allocated or paid time to report the requests. The validity of these data is limited by a poor response rate, a non-validated questionnaire and potential for selection bias. Validation using multiple hospitals and greater numbers is the appropriate next step. Responses suggest that this system identifies areas for improvement, reinforces good practice and improves the mental well-being of the participating clinicians.

Expanding the chemical diversity of M13 bacteriophage.

Bacteriophage M13 virions are very stable nanoparticles that can be modified by chemical and genetic methods. The capsid proteins can be functionalized in a variety of chemical reactions without loss of particle integrity. In addition, Genetic Code Expansion (GCE) permits the introduction of non-canonical amino acids (ncAAs) into displayed peptides and proteins. The incorporation of ncAAs into phage libraries has led to the discovery of high-affinity binders with low nanomolar dissociation constant (K D) values that can potentially serve as inhibitors. This article reviews how bioconjugation and the incorporation of ncAAs during translation have expanded the chemistry of peptides and proteins displayed by M13 virions for a variety of purposes.

Breastfeeding in a Global Context: Epidemiology, Impact, and Future Directions.

INTRODUCTION: More than 98% of infant deaths occur in low- and middle-income countries (LMICs). Breastfeeding improves infant survival and protects against certain illnesses, such as diarrhea and pneumonia, which are leading causes of deaths in those <5 years of age in LMICs. The World Health Organization (WHO) recommends early initiation of breastfeeding within 1 hour of birth, exclusive breastfeeding up to 6 months of age, and continued breastfeeding up to 2 years of age. However, fewer than half of infants in LMICs are breastfed optimally to these standards. The objectives of this article are to describe the global epidemiology and health benefits of breastfeeding with particular focus on LMICs. METHODS: We searched PubMed to identify original research articles on breastfeeding in LMICs and reviews related to the benefits of breastfeeding, with particular focus on articles published in the past 5 years. We used reports and data published by the WHO and the United Nations Children's Fund (UNICEF) related to global breastfeeding rates, targets, and programmatic initiatives. We used the Lives Saved Tool to estimate mortality related to breastfeeding practices. FINDINGS: Less than half of infants globally receive early, exclusive, or continued breastfeeding. Certain high-risk groups, such as premature or HIV-exposed infants, face particular challenges and benefits related to breastfeeding. The WHO, UNICEF, and other global partners have developed a multipronged strategy to promote global breastfeeding, ranging from government-level advocacy to grassroots community support groups. Using the Lives Saved Tool, we estimate that nearly 200,000 lives of those <5 years of age could be saved in LMICs from 2020 to 2030 if early, exclusive, and continued breastfeeding rates were linearly increased from current rates to meet the WHO 2030 goals of 60% to 80% coverage. If this goal were exceeded and near-universal coverage were achieved, the number of lives would increase even further such that >820,000 lives per year could potentially be saved by universal breastfeeding. In this review, we delineate the health and economic benefit of breastfeeding in LMICs, discuss breastfeeding epidemiology in the global context, and describe targeted strategies to improve breastfeeding uptake.

Macrofungi as a Nutraceutical Source: Promising Bioactive Compounds and Market Value.

Macrofungi production and economic value have been increasing globally. The demand for macrofungi has expanded rapidly owing to their popularity among consumers, pleasant taste, and unique flavors. The presence of high quality proteins, polysaccharides, unsaturated fatty acids, minerals, triterpene sterols, and secondary metabolites makes macrofungi an important commodity. Macrofungi are well known for their ability to protect from or cure various health problems, such as immunodeficiency, cancer, inflammation, hypertension, hyperlipidemia, hypercholesterolemia, and obesity. Many studies have demonstrated their medicinal properties, supported by both in vivo and in vitro experimental studies, as well as clinical trials. Numerous bioactive compounds isolated from mushrooms, such as polysaccharides, proteins, fats, phenolic compounds, and vitamins, possess strong bioactivities. Consequently, they can be considered as an important source of nutraceuticals. Numerous edible mushrooms have been studied for their bioactivities, but only a few species have made it to the market. Many species remain to be explored. The converging trends and popularity of eastern herbal medicines, natural/organic food product preference, gut-healthy products, and positive outlook towards sports nutrition are supporting the growth in the medicinal mushroom market. The consumption of medicinal mushrooms as functional food or dietary supplement is expected to markedly increase in the future. The global medicinal mushroom market size is projected to increase by USD 13.88 billion from 2018 to 2022. The global market values of promising bioactive compounds, such as lentinan and lovastatin, are also expected to rise. With such a market growth, mushroom nutraceuticals hold to be very promising in the years to come.

Critical Breaths in Transit: A Review of Non-invasive Ventilation (NIV) for Neonatal and Pediatric Patients During Transportation.

Respiratory illnesses are a leading cause of death for children worldwide, with the majority of these cases occurring from preterm birth complications or acute respiratory infections. Appropriate respiratory intervention must be provided quickly to lower the chances of death or permanent harm. As a result, respiratory support given in prehospital and interfacility transport can substantially improve health outcomes for these patients, particularly in areas where transportation time to appropriate facilities is lengthy. Existing literature supports the use of non-invasive ventilation (NIV), such as nasal or bilevel continuous positive airway pressure, as a safe form of respiratory support for children under 18 years old in certain transportation settings. This mini review summarizes the literature on pediatric NIV in transport and highlights significant gaps that future researchers should address. In particular, we identify the need to: solidify clinical guidelines for the selection of eligible pediatric patients for transport on NIV; explore the range of factors influencing successful NIV implementation during transportation; and apply appropriate best practices in low and middle income countries.

Taxonomy, Diversity and Cultivation of the Oudemansielloid/Xeruloid Taxa Hymenopellis, Mucidula, Oudemansiella, and Xerula with Respect to Their Bioactivities: A Review.

The oudemansielloid/xeruloid taxa Hymenopellis, Mucidula, Oudemansiella, and Xerula are genera of Basidiomycota that constitute an important resource of bioactive compounds. Numerous studies have shown antimicrobial, anti-oxidative, anti-cancer, anti-inflammatory and other bioactivities of their extracts. The bioactive principles can be divided into two major groups: (a) hydrophilic polysaccharides with relatively high molecular weights and (b) low molecular medium polar secondary metabolites, such as the antifungal strobilurins. In this review, we summarize the state of the art on biodiversity, cultivation of the fungi and bioactivities of their secondary metabolites and discuss future applications. Although the strobilurins are well-documented, with commercial applications as agrochemical fungicides, there are also other known compounds from this group that have not yet been well-studied. Polysaccharides, dihydro-citrinone phenol A acid, scalusamides, and acetylenic lactones such as xerulin, also have potential applications in the nutraceutical, pharmaceutical and medicinal market and should be further explored. Further studies are recommended to isolate high quality bioactive compounds and fully understand their modes of action. Given that only few species of oudemansielloid/xeruloid mushrooms have been explored for their production of secondary metabolites, these taxa represent unexplored sources of potentially useful and novel bioactive metabolites.

A bioassay-based protocol for chemical neutralization of human faecal wastes treated by physico-chemical disinfection processes: A case study on benzalkonium chloride.

In situ physico-chemical disinfection of high risk faecal waste is both effective and widely used as a sanitation management strategy for infection prevention and control. Systematic tests where the performance of alternative physico-chemical disinfection methods is systematically compared and optimized must be based on reliable protocols. These protocol are currently not adequately addressing the neutralization related issues: the neutralization of the tested disinfectant after specified conditions of concentration and contact time (CT) is necessary to prevent continued disinfection after the intended contact time; moreover such neutralization is often necessary in practice and on a large scale to prevent adverse health and ecological impacts from remaining disinfectant after the target CT is achieved. Few studies adequately assess the extent of neutralization of the chemical disinfectant and are intended to optimize on-site disinfection practices for waste matrices posing high microbial risks. Hence, there is a need for effective and reproducible neutralization protocols in chemical disinfection trials and practice. Furthermore, for most of chemical disinfectants used in healthcare settings there is no practical methodology to reliably and conveniently measure the residual disinfectant concentration after its neutralization and also determine the optimum concentration of the neutralizer. Because some neutralizing compounds can themselves be toxic to the test microorganisms, it is necessary to optimize neutralization procedures in disinfection experiments for the development of infection control practices using accepted positive control microbes. In the presented work, a stepwise bioassay-based protocol using representative faecal indicator microbes is described for optimizing chemical disinfection and subsequent disinfectant neutralization of any infectious faecal waste matrix. The example described is for the quaternary ammonium compound benzalkonium chloride and its recommended chemical neutralizer in a high strength human faecal waste matrix.