Recent contributions of behavioural and biomedical scientists in applied anthropology of hygiene practices and management of diarrhoea in Bangladesh

In the past few years anthropologists, sociologists and biostatisticians have made contributions on hygiene practices and management of diarrhoea in relation to young children in rural Bangladesh. The cultural practices of caregivers with regard to washing hands after cleansing the anus of young children following defectation and disposing of the stool often do not fulfil the biomedical criteria for prevention of the transmission of diarrhoea-causing pathogens. These practices, being important determinants of prevention of transmission, were studied in the rural communities of Matlab, Bangladesh. Data were collected by observing the practices of caregivers of 404 children with diarrhoea chosen from 444 community cases recruited from the neighbourhood of the 218 hospitalized index cases. Observations were made regarding the cleansing of the anus of these children, washing of the hands of the caregivers, and disposal of the stool. It was observed that all the caregivers used bare hands and most used only water in cleaning the anus of the child. Also following the cleaning, most of the caregivers used only water to wash the soiled hand. Most of them used water collected in a pot, and only a small proportion used water from the direct source. The majority of caregivers disposed of the faeces in an unhygienic way. The caregivers classified diarrhoeal illness into 5 types based on stool characteristics and perceived specific causation. Their management approach was determined by the type and cause of the diarrhoea. The oral rehydration approach to management was not considered for diarrhoeas which were believed to be caused by supernatural and certain other cultural factors.     

The need to incorporate human variation and evolutionary theory in forensic anthropology: A call for reform

In 1992, Norm Sauer called for a language shift in which practitioners would move away from the socially loaded term “race” and replace it with the less provocative term “ancestry.” While many heeded the call and moved towards ancestry in their research and reports, the actual approach to research and analysis did not change. In response to this change, there was a large growth in ancestry estimation method development in the early decade of the 2000s. However, the practice of ancestry estimation did not adequately incorporate evolutionary theory in interpretation or trait selection and continued with little critical reflection. In the past decade, there has been an increase in ancestry validation methods with little critique of the “race” concept or discussion of modern human variation or reference samples. To advance, forensic anthropologists need to reckon with the practice of ancestry estimation as it is currently practiced. We are calling for another reform in the axiom focusing on evolutionary theory, population history, trait selection, and population-level reference samples. The practice needs to abandon the terms ancestry and race completely and recalibrate to an analysis of population affinity. Population affinity is a statistical approach based on the underlying population structure that would allow the understanding of how microevolutionary forces act in concert with historical events (e.g., colonization, the Transatlantic Slave Trade, etc.) to shape modern human variation. This is not to be confused with geographic ancestry that all too often can be perceived as interchangeable with social race and as an affirmation of the biological concept of race. It is time to critically evaluate the social and scientific implications of the current practice of ancestry estimation, and re-frame our approach to studying and analyzing modern human variation through a population structure approach.     

Seeking consilience: Traditional ecological knowledge and Western social science contributions to orca conservation knowledge

Consilience is the integration of disciplinary knowledges in search of a more complete truth. In the complex context of conservation, where human activities are increasingly impacting the population status of many species, this endeavor is particularly important. Yet, to date, we have had limited attempts at unifying diverse sources of knowledge around a conservation issue. Focusing on orca conservation specifically, we share the perspectives of five scholars from five disciplines to demonstrate how Indigenous Knowledges and Social Sciences can inform the conservation of Southern Resident Killer Whales (SRKWs). We see Traditional Ecological Knowledge (TEK) as an original consilient knowledge and Western social sciences as the fields that can best identify and describe the norms and patterns of how to engage conservation. The integration of these broader knowledge systems, driven by individuals trained in their respective fields, with the already existing biophysical data around SRKWs, can help us make better decisions for SRKW conservation.     

Tissue contributions to sex and race: differences in tooth crown size of deciduous molars.

This study describes size of constituent deciduous tooth crown components (enamel, dentine, and pulp) to address the manner in which males characteristically have larger teeth than females, and the observation that teeth of American blacks are larger than those of American whites. Measurements were collected (n = 333 individuals) from bitewing radiographs using computer-aided image analysis. Tissue thicknesses (enamel, dentine, pulp) were measured at the crown's mesial and distal heights of contour. Deciduous mesiodistal molar crown length is composed of about 1/7 enamel, 1/3 dentine, and 1/2 pulp. Details differ by tooth type, but males typically have significantly larger dentine and pulp dimensions than females; there is no sexual dimorphism in marginal enamel thickness. Males scale isometrically with females for all variables tested here. Blacks significantly exceed whites in size of all tissues, but tissue types scale isometrically with blacks and whites with one exception: enamel thickness is disproportionately thick in blacks. While the absolute difference is small (5.56 mm of enamel in blacks summed over all four deciduous molar tooth types vs. 5.04 mm in whites), the statistical difference is considerable (P < 0.001). Aside from enamel, crown size in blacks is increased proportionately vis-à-vis whites. Principal components analysis confirmed these univariate relationships and emphasizes the statistical independence of crown component thicknesses, which is in keeping with the sequential growth and separate embryonic origins of the tissues contributing to a tooth crown. Results direct attention to the rates of enamel and dentine deposition (of which little is known), since the literature suggests that blacks (with larger crowns and thicker enamel) spend less time in tooth formation than whites.     

开展医疗器械安全工程学研究防止不良事件发生

主要介绍了医疗器械安全工程学。近年来日本、欧美等发达国家积极开展这个学科的研究,以减少不良事件的发生。除一般的安全问题外,医疗器械安全工程学研究的主要内容包括人因工程、人体工程学、医用软件、人机接口及误使用等。     

Using engineering models to shorten cryoprotectant loading time for the vitrification of articular cartilage

Osteochondral allograft transplantation can treat full thickness cartilage and bone lesions in the knee and other joints, but the lack of widespread articular cartilage banking limits the quantity of cartilage available for size and contour matching. To address the limited availability of cartilage, vitrification can be used to store harvested joint tissues indefinitely. Our group's reported vitrification protocol [Biomaterials 33 (2012) 6061–6068] takes 9.5 h to load cryoprotectants into intact articular cartilage on bone and achieves high cell viability, but further optimization is needed to shorten this protocol for clinical use. Herein, we use engineering models to calculate the spatial and temporal distributions of cryoprotectant concentration, solution vitrifiability, and freezing point for each step of the 9.5-h protocol. We then incorporate the following major design choices for developing a new shorter protocol: (i) all cryoprotectant loading solution concentrations are reduced, (ii) glycerol is removed as a cryoprotectant, and (iii) an equilibration step is introduced to flatten the final cryoprotectant concentration profiles. We also use a new criterion—the spatially and temporally resolved prediction of solution vitrifiability—to assess whether a protocol will be successful instead of requiring that each cryoprotectant individually reaches a certain concentration. A total cryoprotectant loading time of 7 h is targeted, and our new 7-h protocol is predicted to achieve a level of vitrifiability comparable to the proven 9.5-h protocol throughout the cartilage thickness.     

The contributions of aspirin and microbial oxygenase to the biosynthesis of anti-inflammatory resolvins: novel oxygenase products from omega-3 polyunsaturated fatty acids

Resolvins (Rvs) are oxygenated products derived from omega-3 polyunsaturated fatty acids (PUFAs) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid that carry potent protective bioactions present in resolving inflammatory exudates. Resolvin E1 (RvE1) is biosynthesized in vivo from EPA via transcellular biosynthetic routes during cell-cell interactions, and thus RvE1 is formed in vivo during multicellular responses such as inflammation and microbial infections. RvE1 protects tissues from leukocyte-mediated injury and counterregulates proinflammatory gene expression. These newly identified Rvs may underlie the beneficial actions of omega-3 PUFAs especially in chronic disorders where unresolved inflammation is a key mechanism of pathogenesis. Here, we present an overview of the biosynthesis of RvE1, with a focus on the aspirin-triggered and microbial P450-initiated pathways. The generation of RvE1 and its actions appear to dampen acute leukocyte responses and facilitate the resolution of inflammation.     

Using GaBi 3 to perform life cycle assessment and life cycle engineering

The growing availability of software tools has increased the speed of generating LCA studies. Databases and visual tools for constructing material balance modules greatly facilitate the process of analyzing the environmental aspects of product systems over their life cycle. A robust software tool, containing a large LCI dataset and functions for performing LCIA and sensitivity analysis will allow companies and LCA practitioners to conduct systems analyses efficiently and reliably. This paper discusses how the GaBi 3 software tool can be used to perform LCA and Life Cycle Engineering (LCE), a methodology that combines life cycle economic, environmental, and technology assessment. The paper highlights important attributes of LCA software tools, including high quality, well-documented data, transparency in modeling, and data analysis functionality. An example of a regional power grid mix model is used to illustrate the versatility of GaBi 3.     

High throughput engineering to revitalize a vestigial electron transfer pathway in bacterial photosynthetic reaction centers

Photosynthetic reaction centers convert light energy into chemical energy in a series of transmembrane electron transfer reactions, each with near 100% yield. The structures of reaction centers reveal two symmetry-related branches of cofactors (denoted A and B) that are functionally asymmetric; purple bacterial reaction centers use the A pathway exclusively. Previously, site-specific mutagenesis has yielded reaction centers capable of transmembrane charge separation solely via the B branch cofactors, but the best overall electron transfer yields are still low. In an attempt to better realize the architectural and energetic factors that underlie the directionality and yields of electron transfer, sites within the protein-cofactor complex were targeted in a directed molecular evolution strategy that implements streamlined mutagenesis and high throughput spectroscopic screening. The polycistronic approach enables efficient construction and expression of a large number of variants of a heteroligomeric complex that has two intimately regulated subunits with high sequence similarity, common features of many prokaryotic and eukaryotic transmembrane protein assemblies. The strategy has succeeded in the discovery of several mutant reaction centers with increased efficiency of the B pathway; they carry multiple substitutions that have not been explored or linked using traditional approaches. This work expands our understanding of the structure-function relationships that dictate the efficiency of biological energy-conversion reactions, concepts that will aid the design of bio-inspired assemblies capable of both efficient charge separation and charge stabilization.     

Human internal mammary artery (IMA) transplantation and stenting: a human model to study the development of in-stent restenosis

Preclinical in vivo research models to investigate pathobiological and pathophysiological processes in the development of intimal hyperplasia after vessel stenting are crucial for translational approaches (1,2). The commonly used animal models include mice, rats, rabbits, and pigs (3-5). However, the translation of these models into clinical settings remains difficult, since those biological processes are already studied in animal vessels but never performed before in human research models (6,7). In this video we demonstrate a new humanized model to overcome this translational gap. The shown procedure is reproducible, easy, and fast to perform and is suitable to study the development of intimal hyperplasia and the applicability of diverse stents. This video shows how to perform the stent technique in human vessels followed by transplantation into immunodeficient rats, and identifies the origin of proliferating cells as human.     

A poor imitation of a natural process: A call to reconsider the iPSC engineering technique

Reprogramming somatic cells into a pluripotent state is expected to initiate a new era in medicine. Because the precise underlying mechanism of reprogramming remains unclear, many efforts have been made to optimize induced pluripotent stem cell (iPSC) engineering. However, satisfactory results have not yet been attained. In this review, we focus on recent roadblocks in iPSC reprogramming engineering, such as the inefficiency of the process, tumorigenicity and heterogeneity of the generation. We conclude that cell reprogramming is a naturally occurring phenomenon rather than a biological technique. We will only be able to mimic the natural process of reprogramming when we fully understand its underlying mechanism. Finally, we highlight the alternative method of direct conversion, which avoids the use of iPSCs to generate cell materials for patient-specific cell therapy.     

Affinity maturation to improve human monoclonal antibody neutralization potency and breadth against hepatitis C virus

A potent neutralizing antibody to a conserved hepatitis C virus (HCV) epitope might overcome its extreme variability, allowing immunotherapy. The human monoclonal antibody HC-1 recognizes a conformational epitope on the HCV E2 glycoprotein. Previous studies showed that HC-1 neutralizes most HCV genotypes but has modest potency. To improve neutralization, we affinity-matured HC-1 by constructing a library of yeast-displayed HC-1 single chain Fv (scFv) mutants, using for selection an E2 antigen from one of the poorly neutralized HCVpp. We developed an approach by parallel mutagenesis of the heavy chain variable (VH) and κ-chain variable (Vk) genes separately, then combining the optimized VH and Vk mutants. This resulted in the generation of HC-1-related scFv variants exhibiting improved affinities. The best scFv variant had a 92-fold improved affinity. After conversion to IgG1, some of the antibodies exhibited a 30-fold improvement in neutralization activity. Both surface plasmon resonance and solution kinetic exclusion analysis showed that the increase in affinity was largely due to a lowering of the dissociation rate constant, Koff. Neutralization against a panel of HCV pseudoparticles and infectious 2a HCV virus improved with the affinity-matured IgG1 antibodies. Interestingly, some of these antibodies neutralized a viral isolate that was not neutralized by wild-type HC-1. Moreover, propagating 2a HCVcc under the selective pressure of WT HC-1 or affinity-matured HC-1 antibodies yielded no viral escape mutants and, with the affinity-matured IgG1, needed 100-fold less antibody to achieve complete virus elimination. Taken together, these findings suggest that affinity-matured HC-1 antibodies are excellent candidates for therapeutic development.     

A Novel Three-dimensional Flow Chamber Device to Study Chemokine-directed Extravasation of Cells Circulating under Physiological Flow Conditions

Extravasation of circulating cells from the bloodstream plays a central role in many physiological and pathophysiological processes, including stem cell homing and tumor metastasis. The three-dimensional flow chamber device (hereafter the 3D device) is a novel in vitro technology that recreates physiological shear stress and allows each step of the cell extravasation cascade to be quantified. The 3D device consists of an upper compartment in which the cells of interest circulate under shear stress, and a lower compartment of static wells that contain the chemoattractants of interest. The two compartments are separated by porous inserts coated with a monolayer of endothelial cells (EC). An optional second insert with microenvironmental cells of interest can be placed immediately beneath the EC layer. A gas exchange unit allows the optimal CO₂ tension to be maintained and provides an access point to add or withdraw cells or compounds during the experiment. The test cells circulate in the upper compartment at the desired shear stress (flow rate) controlled by a peristaltic pump. At the end of the experiment, the circulating and migrated cells are collected for further analyses. The 3D device can be used to examine cell rolling on and adhesion to EC under shear stress, transmigration in response to chemokine gradients, resistance to shear stress, cluster formation, and cell survival. In addition, the optional second insert allows the effects of crosstalk between EC and microenvironmental cells to be examined. The translational applications of the 3D device include testing of drug candidates that target cell migration and predicting the in vivo behavior of cells after intravenous injection. Thus, the novel 3D device is a versatile and inexpensive tool to study the molecular mechanisms that mediate cellular extravasation.     

Adenoviral Transduction of Naive CD4 T Cells to Study Treg Differentiation

Regulatory T cells (Tregs) are essential to provide immune tolerance to self as well as to certain foreign antigens. Tregs can be generated from naive CD4 T cells in vitro with TCR- and co-stimulation in the presence of TGFβ and IL-2. This bears enormous potential for future therapies, however, the molecules and signaling pathways that control differentiation are largely unknown.Primary T cells can be manipulated through ectopic gene expression, but common methods fail to target the most important naive state of the T cell prior to primary antigen recognition. Here, we provide a protocol to express ectopic genes in naive CD4 T cells in vitro before inducing Treg differentiation. It applies transduction with the replication-deficient adenovirus and explains its generation and production. The adenovirus can take up large inserts (up to 7 kb) and can be equipped with promoters to achieve high and transient overexpression in T cells. It effectively transduces naive mouse T cells if they express a transgenic Coxsackie adenovirus receptor (CAR). Importantly, after infection the T cells remain naive (CD44^low, CD62L^high) and resting (CD25^-, CD69^-) and can be activated and differentiated into Tregs similar to non-infected cells. Thus, this method enables manipulation of CD4 T cell differentiation from its very beginning. It ensures that ectopic gene expression is already in place when early signaling events of the initial TCR stimulation induces cellular changes that eventually lead into Treg differentiation.