Tupaia belangeri, as well as HepaRG cells, important determinants of viral entry have been achieved. Recently, sodium taurocholate cotransporting polypeptide (NTCP) was identified as an HBV entry receptor and enabled the establishment of a susceptible cell line that can efficiently support HBV infection. This finding will allow a deeper understanding of the requirements for efficient HBV infection, including the elucidation of the molecular entry mechanism. In addition, pharmacological studies suggest that NTCP is able to serve as a therapeutic target. This article summarizes our current knowledge on the mechanisms of HBV entry and the role of NTCP in this process.Chronic hepatitis B virus (HBV) infection, affecting approximately 240 million people worldwide, is a major public health problem that elevates the risk of developing liver cirrhosis and hepatocellular carcinoma. Given that current anti-HBV drugs are limited to interferon-based regimens and nucleos(t)ide analogs, the development of new anti-HBV agents is urgently needed. The viral entry process is generally an attractive target implicated in antiviral strategies. Using primary cells from humans and  ChronicIn general, the viral entry step is an attractive target for the development of antiviral agents –8. The e2.HBV infection into host hepatocytes follows a multiple step process: (1) initially, HBV reversibly attaches to host cell surface proteoglycans with a low affinity; (2) this is followed by the process involving more specific receptor(s) with high affinity to mediate the early entry step; and (3) after endocytosis-mediated internalization, the virus fuses with the cellular membrane compartment, probably in an endosomal compartment, although the mechanisms are not fully understood. Both initial attachment and, probably more importantly, specific receptor recognition contribute to host specificity and tissue tropism . The iniTupaia belangeri, as well as HepaRG cells . Th. Thin viitro (50 ,81, furtAs other examples, compounds known to be NTCP inhibitors, including progesterone, propranolol and bosentan, have been shown to block HBV infection  42]. NT. NT42]. Host-targeting antivirals are generally expected to have significant advantages, including a much lower frequency drug resistance, universal antiviral effects beyond viral genotypes and complementary mechanisms of action that might act in a synergistic manner with currently available antiviral agents . More im7.in vivo small animal model that fully supports HBV infection.Identification of NTCP as an HBV entry receptor has accelerated the understanding of HBV molecular biology and offered useful experimental systems to analyze the HBV and HDV lifecycle, including the identification of host restriction and dependency factors. NTCP also represents a new therapeutic target in the development of new anti-HBV agents. Further analyses using a new cell culture system are necessary in order to clarify the molecular mechanisms underlying NTCP-mediated HBV infection and to establish an


Fractional exhaled nitric oxide (FeNO) is a non invasive method for assessing the inflammatory status of children with airway disease. Different ways to measure FeNO levels are currently available. The possibility of measuring FeNO levels in an office setting even in young children, and the commercial availability of portable devices, support the routine use of FeNO determination in the daily pediatric practice. Although many confounding factors may affect its measurement, FeNO is now widely used in the management of children with asthma, and seems to provide significantly higher diagnostic accuracy than lung function or bronchial challenge tests. The role of FeNO in airway infection , in bronchiectasis, or in cases with diffuse lung disease is less clear. This review focuses on the most recent advances and the current clinical applications of FeNO measurement in pediatric lung disease. Nitric oxide (NO) is a biological mediator synthesized by NO synthase (NOS), an enzyme that catalyzes the oxidation of L-arginine to NO and L-citrulline. Constitutive NOS isoenzymes include neuronal NOS NOS1) and endothelial NOS NOS3), both of which are activated by calcium ions to produce small amounts of NO  and endo. NO was , both ofDuring the last years the availability of NO analyzers and the publication of official guidelines made the measurement of fractional exhaled NO (FeNO) a useful tool in the diagnosis of various pediatric airway disorders -10. This2O oral pressure. NO concentrations in balloons can be stable for several hours, and the measurement can take place also at a distant site. Flow rate standardization improves the reproducibility of the offline technique, with results similar to the online constant flow rate methods [Several methods are used to measure FeNO in children, and the choice depends on the subject’s age and cooperation. Online measurement allows FeNO testing with a real-time display of NO breath profiles, which is available in stationary devices only. Offline testing, instead, refers to collection of exhaled air into suitable receptacles for delayed analysis . The sin methods . In subj methods . The chi methods .Finally, the agreement between different devices gave inconsistent results, with some studies showing significant differences between analyzers , while oSeveral recent publications have reported reference values for FeNO in children and adolescents -28 Tabl. There aChronic airway inflammation is an important feature in the development and progression of bronchial asthma ,33-36. FObesity is another confounding factor in the assessment of FeNO levels. A consistent body of data now indicates that asthma is closely related to obesity, and obese patients with asthma usually have more severe symptoms than non-obese asthmatics . Indeed,As far as the discrimination between children with asthma and healthy subjects, FeNO measurement provides significantly higher diagnostic accuracy than lung function tests ,73, and Cut-points rather than reference values have been proposed to interpret FeNO levels ,34,72,79An additional, novel, potential and attractive application of FeNO in asthma is the “prediction” of asthma onset. In the absence of symptoms, increased FeNO levels may reflect subclinical airway inflammation that may be predictive of “early asthma”, especially in allergic subjects ,82. ThisThe goal of asthma long-term treatment is to reduce inflammation for controlling symptoms. Treatment options are usually guided by symptoms and lung function. However, these factors do not reflect chronic airway inflammation. This is also shown by contrasting results of studies on the relationship between FeNO levels and both symptoms , symptom control, or rescue ß2-agonist use ,85-94 anet al showed that the addition of FeNO measurement as an indicator of asthma control resulted in higher doses of ICS and long-acting β2 agonists than did standard guideline-based treatment, and did not determine improvements in asthma symptoms or lung function [Inhaled corticosteroids (ICS) are the first choice for asthma maintenance treatment. Interestingly, inhaled or systemic corticosteroid administration results in dose dependent reductions of FeNO levels ,100. Morfunction . A recenfunction .These findings may be explained by the fact that day-to-day variations of FeNO are common and do not correlate with changes in symptom score . FurtherAnother issue is whether the change in FeNO values may be a better predictor than absolute levels. FeNO levels quickly decrease in response to ICS and therefore they may be useful to ascertain that ICS is regularly taken. Finally, baseline FeNO seems a worse predictor of asthma improvement than the change in FeNO after 80 days of ICS .According to the Clinical Practice Guideline of the American Thoracic Society (ATS)  it has bet al assessed the value of spirometry and FeNO measurements, alone or in combination, in models developed by a machine learning approach for the objective classification of asthma control [et al found that FeNO measured daily by a hand-held device started to increase approximately 10 days before moderate exacerbations occurred, this suggesting that regular FeNO measurements in the home setting could help to detect and even to prevent the loss of asthma control [The results of two recent studies indicate new possible clinical applications of FeNO measurement in pediatric asthma. Pifferi  control . The com control . Apart f control -117.On the basis of the studies that have provided evidence regarding the applications of NO measurements in clinical practice, ATS recently indicated the rationale for FeNO measurement in asthma, even in the pediatric population , highlig– Diagnosis of eosinophilic airway inflammation– Support of asthma diagnosis when objective evidence is needed– Baseline evaluation and follow-up monitoring of airway inflammation– Assessment of potential response or failure to respond to inhaled corticosteroids– Evaluation of adherence to antiinflammatory medications– Guide for dose changes in antiinflammatory medicationsRespiratory Syncytial Virus (RSV) manifests restricted tropism for the respiratory epithelium stimulating an inflammatory response . RSV incThe scientific literature on FeNO modifications during community acquired pneumonia (CAP) in children is scarce. Since NO is part of the innate inflammatory response, its levels are expected to rise in acute lung infection. Healthy children with an abnormally high FeNO had significantly increased frequency of previous bronchitis or pneumonia in the past year . HoweverBronchiectasis is caused by, or associated with, many disorders including congenital/genetic conditions, e.g. cystic fibrosis (CF), primary immunodeficiency, primary ciliary dyskinesia (PCD), Mounier-Kuhn syndrome, chronic obstructive pulmonary disease, bronchiolitis obliterans, sarcoidosis, autoimmune disorders, and acquired post-infectious diseases -131. HigLevels of inflammation in stable bronchiectasis seem to correlate with a reduction of patient’s quality of life . TherefoNO measurement at different expiratory flow rates allows to assess the contribution of NO from different parts of the lung. A two-compartment model of pulmonary NO exchange dynamics has been proposed to demonstrate relative contributions of bronchial (J) and peripheral  airway NO to the final FeNO concentration . In adulNotwithstanding the conflicting results between affected children and adults, FeNO measurement could represent a useful non invasive tool to monitor bronchial inflammation over time.The term diffuse lung disease (DLD) encompasses a heterogeneous group of chronic respiratory disorders characterized by abnormal gas exchange and diffuse radiographic and histopathologic abnormalities . DLD is Several studies evaluated FeNO levels in adults with DLD. In systemic sclerosis, FeNO concentrations are lower in subjects with interstitial lung involvement than in those without, whereas patients without pulmonary disease have higher FeNO than healthy subjects . MoreoveBronchiolitis obliterans (BO) is almost secondary to lung transplantation in adults , while tNO seems to significantly influence a variety of physiological and pathophysiological processes in the upper and lower airways. FeNO measurement is easy to perform, and seems reproducible also in the preschool age. The availability of hand-held devices will hopefully increase its use in the pediatric practice. Combined with symptoms registration and lung function measurements, FeNO provides additional information that can be applied to support the diagnosis of asthma and to optimize the management of affected children. Moreover, recent studies suggest that FeNO is helpful in predicting both onset and exacerbations of asthma. Since most studies have been performed in adult populations, more research is needed to confirm the usefulness of NO measurement in the diagnosis and management of pediatric chronic airway disorders different from asthma.NO: Nitric oxide; NOS: NO synthase; FeNO: Fractional exhaled nitric oxide; ICS: Inhaled corticosteroids; RSV: Respiratory syncytial virus; CAP: Community acquired pneumonia; CF: Cystic fibrosis, PCD, Primary ciliary dyskinesia; HRCT: High-resolution computed tomography; DLD: Diffuse lung disease; BAL: Bronchoalveolar lavage; BO: Bronchiolitis obliterans.All authors declare that they have no significant competing financial, professional or personal interests that might have influenced the performance or presentation of the work described in this manuscript.AM has been involved in drafting the manuscript; CC has been involved in revising the manuscript critically for important intellectual content; MV has made substantial contributions to acquisition, analysis and interpretation of data; CP has made substantial contributions to acquisition, analysis and interpretation of data; SM has been involved in drafting the manuscript; MM has been involved in drafting the manuscript; FSp has made substantial contributions to acquisition, analysis and interpretation of data; FS has been involved in drafting the manuscript, revising it critically for important intellectual content and has given final approval of the version to be published. All authors read and approved the final manuscript.


Positive-sense RNA viruses are responsible for frequent and often devastating diseases in humans, animals, and plants. However, the development of effective vaccines and anti-viral therapies targeted towards these pathogens has been hindered by an incomplete understanding of the molecular mechanisms involved in viral replication. One common feature of all positive-sense RNA viruses is the manipulation of host intracellular membranes for the assembly of functional viral RNA replication complexes. This review will discuss the interplay between cellular membranes and positive-sense RNA virus replication, and will focus specifically on the potential structural and functional roles for cellular lipids in this process. One group of viruses responsible for a wide range of diseases in humans, animals, and plants are classified as positive-sense RNA viruses due to their genome structure, which consists of one or more single-stranded RNA molecules that in many respects resemble cellular mRNAs. Clinically important members of this group cause significant morbidity and mortality, include viruses from the  threats ,2. Curre threats ,4. An al threats –9. Howevin vitro [in vitro viral replicase activity [The small genomes of viruses relative to other organisms requires that they appropriate cellular machinery to complete their replication cycle, which for positive-sense RNA viruses is depicted schematically in in vitro ,28,40–45activity –45. And activity –63.et al. forming part of this special issue of Viruses.Although the observations outlined above provide substantial validation for the important role that cellular membranes play in positive-sense RNA virus replication, they provide only circumstantial evidence for the specific membrane components involved in the process and their precise molecular functions with respect to viral genome replication. Intracellular membranes contain diverse protein and lipid constituents and play a variety of roles during normal cellular physiology and metabolism, which include facilitating the spatial separation of cellular processes and the consequent differential concentrations of crucial cellular components, providing structural integrity to maintain organelle shape, and contributing functional co-factors for multiple processes such as signal transduction and biosynthesis. In this review, we will discuss three potential roles that lipid constituents of host cell membranes play in positive-sense RNA virus genome replication. These include: (i) providing a scaffold for targeting and assembly of RNA replication complexes; (ii) inducing alterations in membranes structure to potentially shield viral RNA replication intermediates from cellular innate antiviral pathways; and (iii) serving as functional co-factors for optimal enzymatic activity of viral replicase proteins. We will use examples of plant, insect, and mammalian viruses to highlight specific aspects of these potential roles, and will focus primarily on lipids rather than membranes in general. Several excellent reviews have recently been published –14 for r1.1.The interior of a cell is a highly organized environment, where membrane-bound organelles and specific organelle membranes represent one mechanism whereby cells enforce spatial constraints on particular metabolic processes. For example, the enzymes involved in electron transport and respiration are concentrated on the inner mitochondria membrane. Similarly, viruses may use intracellular membranes as simple molecular scaffolds on which to assemble their replication complexes, thereby increasing replication efficiency by concentrating essential viral and potential cellular components within a smaller microenvironment in the cell. Furthermore, membranes may provide a convenient platform to coordinate various steps in the replication cycle see , such as1.2.Positive-sense RNA viruses induce a number of distinctive membrane structures, which include the “membranous webs” of hepatitis C virus , the cluThe structural changes that must occur within lipid bilayers to produce the virus-induced alterations in intracellular membranes are substantial. For example, the outer mitochondrial membrane invaginations induced by Flock House virus  see als and the An alternative mechanism to induce membrane curvature is modification of lipid structure, either through changes in the polar headgroup or acyl chain composition. For example, lysophospholipids, which contain only one acyl chain per phospholipid molecule, and special lipids such as cholesterol or cardiolipin, which has four acyl chains attached to diphosphatidylglycerol, can have profound impacts on membrane curvature and plasticity . Acyl chRegardless of the mechanism whereby membrane curvature occurs, the function of these curious virus-induced structures remains enigmatic. One interesting possibility is that virus-induced membrane structures shield viral products that have the potential to activate cellular innate immune pathways. The replication of positive-sense RNA viruses involves the formation of dsRNA intermediates that serve as templates for genome amplification (see below, 1.3.de novo synthesis of a functional RNA replication complex in cell-free extracts has been accomplished for both poliovirus [One major goal of positive-sense RNA virus research is the complete isolation, characterization, and de novo synthesis of fully functional RNA replication complexes, a generic example of which is shown schematically in liovirus  and seveliovirus , and thiA substantial focus in the field of positive-sense RNA virus research has been the identification and characterization of the protein components necessary for replication complex function. The impact of lipids on this process has received far less attention in the past, in part due to some of the technical difficulties in lipid research noted above. However, there is both circumstantial and direct evidence that lipids can serve as functional co-factors for viral replicase proteins ,28,40–452.Despite the axiom that cellular membranes are essential host factors for positive-sense RNA virus replication, the specific role of individual membrane components, and in particular lipids, represents a vastly understudied area of virus biology and pathogenesis. We have discussed three potential structural or functional roles that cellular membrane-resident lipids may play in the assembly and function of positive-sense RNA virus replication complexes. These roles are not mutually exclusive, and it is possible that lipids contribute to several steps in the virus life cycle via multiple mechanisms, some of which we currently recognize, and others that remain to be discovered. The recent increase in targeted and genome-wide screens to identify host factors that impact positive-sense RNA virus replication –63, seve


It has been well documented that aging is associated with declines in a variety of cognitive functions. A growing body of research shows that the age-related cognitive declines are reversible through cognitive training programs, suggesting maintained cognitive plasticity of the aging brain. Retest learning represents a basic form of cognitive plasticity. It has been consistently demonstrated for adults in young-old and old-old ages. Accumulated research indicates that retest learning is effective, robust, endurable and could occur at a more conceptual level beyond item-specific memorization. Recent studies also demonstrate promisingly broader transfer effects from retest practice of activities involving complex executive functioning to other untrained tasks. The results shed light on the development of self-guided mental exercise programs to improve cognitive performance and efficiency of the aging brain. The relevant studies were reviewed, and the findings were discussed in light of their limitations, implications, and future directions. In research on cognition and aging, cognitive declines among older adults have been commonly documented in both cross-sectional  and lonA large body of cognitive training studies has focused on directly teaching older adults strategies, including mnemonic techniques , cognitAlternatively, an emerging body of studies adopted a process-specific training approach to examine the benefits of retest practice of psychometric tests of fluid ability , executive functions , cognitively stimulating activities  on older adults' performance on the trained or other untrained cognitive tasks  Does retest learning benefit older adults? (2) Is retest learning endurable in old age? (3) Is retest learning item-specific? (4) Can older adults transfer retest learning to untrained tasks? At the end, the results would be discussed in light of their limitations and implications, together with some proposed future research directions.Data from longitudinal studies revealed substantial retest effects in cognitive abilities among adults aged 18–70 e.g., –32). Ech. Ech32])Earlier studies on retest learning mainly focused on improving performance on tasks measuring fluid intelligence or cognitive mechanics, such as speed and reasoning e.g., , 34). Ho. Ho34]).To investigate whether the aforementioned retest learning could be extended into oldest-old adults in their 80s and over as well as whether the magnitude of retest learning is moderated by age and cognitive functioning status, Yang et al.  comparedRetest practice paradigm has also been employed in some recent studies that aimed to improve older adults' working memory and executive functions , 35–37).–37.35–37In sum, training studies with a practice-oriented retest paradigm on fluid psychometric intelligence (such as reasoning and speed) and executive functioning  consistently revealed substantial improvement in older adults, suggesting that older adults reserve the basic form of cognitive plasticity.In most studies involving retest learning, particularly those longitudinal or training studies on fluid intelligence, participants are repeatedly tested with identical items/tasks across the sessions. It is possible that the retest learning effects may be primarily driven by item-specific effects through memorization/familiarization with specific items and/or solutions. It has been argued that age effects observed in longitudinal studies are composed of positive effects associated with repeated retest practice  and negative effects associated with age e.g., , 32). Ho. Ho32]).To investigate whether older adults show retest learning in the absence of item-specific effects, Yang and colleagues  specificIn studies with strategy-based cognitive training, it has been shown that older adults not only improve their cognitive performance through cognitive training but also maintain the training-induced improvement for a long period of time, such as up to 7 years , 44. EchHowever, to our knowledge, only a few studies have examined the long-term maintenance of retest learning effects in cognitive plasticity studies with older adults. Li et al.  investigIn the domain of fluid ability , transfer effects from either strategy-based training or practice-oriented retest training are very limited and highly ability specific , 19. TraRetest learning studies on executive functioning produced promising yet mixed evidence in terms of transfer effects. Similar to the training studies in fluid ability, a number of studies have demonstrated near transfer effects  of practice with the core process, such as executive functioning or working memory performance. In the study of Dahlin et al. , both yoPromisingly, a growing number of studies revealed broader far transfer effects  from the intervention programs that target at practicing/exercising fundamental core executive functioning using multiple tasks (or multiple versions of the same task) and with complex video games. In the task-switching training study by Karbach and Kray , all ageRecently, an emerging body of studies suggested that participation in cognitively stimulating activities or immersion in an engaged life style , even for a short period of time, could significantly enhance fluid ability, flexible thinking, executive functions, and memory performance –51. For Taken together, relative to retest practice of fluid ability, retest practice of executive functioning or activities involving executive control produced encouraging far transfer effects. This is probably because the executive functioning training focuses on nonspecific fundamental processes underlying performance on a broad range of cognitive tasks. In support to this assumption, a large-scale study by Smith and colleagues  with comIn addition, there has been empirical evidence suggesting that executive functioning is largely regulated by prefrontal cortex , a brainInspired by promisingly broad transfer effects emerged from retest practice of executive functions or cognitively engaged life style, the following factors/features may potentially promote transfer effects: (1) the trained tasks and the transfer tasks share some commonalities ; (2) a focus on exercising fundamental functioning or improving basic cognitive efficiency ; (3) engagement of cognitively stimulating activities involving executive control and promoting self-regulation ; (4) an extensive practice schedule lasting at least for weeks.Taken together, like most classical cognitive training studies , the reThe inspiring findings reviewed above should be evaluated in light of limitations commonly observed in retest learning studies. First, most of the studies involve a highly selected sample that features high-functioning and well-educated older adults, thus we need to be careful to avoid overgeneralization of the results to low-functioning older population. Second, the exact mechanisms underlying the retest improvement are still poorly understood, although previous studies shed a light on potential contributions of item-specific memorization, self-discovered strategies with practice, familiarity with testing situation, and implicit procedural learning , 41. It Despite the limitations, retest learning literature revealed significant and robust retest learning improvement in the performance of practiced tasks in older adults, even in oldest-old age. A growing body of studies suggests that interventions with practicing on activities engaging complex executive functioning have promising broad transfer effects to benefit older adults' general cognitive function. These findings provide empirical foundation for practical application of the development of self-guided cognitive retest programs that could be integrated into older adults' everyday life. In comparison with most trainer-guided and strategy-based training programs, retest learning program is most economic and convenient for older adults because it does not require a trainer and it does not necessarily follow a strict schedule. Older adults could do it at their convenient time and location. All they need is a well-designed program (either a computerized program or a series of booklets) that targets the complex executive functioning and cognitive efficiency  of central neural system. This approach has been implemented in some recent work, although their benefits on older adults' everyday functions are still unclear. For example, Schmiedek and colleagues  developeIn light of promising broad transfer effects of retest practice on tasks heavily loaded with requirement in frontal-lobe-dependent executive control functioning, future studies should expand and explore the optimal conditions, features, and scopes of training to be most beneficial to the daily functions of older adults. This is a challenging direction given the overall limited and task-specific transfer effect revealed in most cognitive training studies, including both strategy-based or retest learning paradigm. But it is extremely meaningful because the ultimate goal of training/practice is to improve older adults' daily functioning and reduce their independence. Future studies should also be directed to the beneficial effects of life-long practice of an engaged life-style or mental exercise materials/programs in improving older adults' cognitive functioning.The existing retest learning and aging literature suggests that older adults, even those in advanced ages, reserve substantial basic form of cognitive plasticity. They demonstrate sizable retest learning benefits from practicing on tasks measuring fluid ability and executive functioning. The retest learning effects are substantial, robust, and endurable and could occur at a non-item-specific conceptual level. In the domain of fluid intelligence, older adults are also capable of transferring retest learning to other ability-specific tasks, with equivalent transfer effects from self-guided retest practice and tutor-guided training. Some recent studies show promisingly broad transfer effects from retest practice of tasks engaging executive functioning. Practice of executive functioning provides a valuable and promising self-guided intervention approach for enhancing older adults' brain functions in prefrontal cortex, general cognitive functioning, and hopefully daily activities and functions.


Dear Editor,We read with great interest the article by Malatesta-Muncher et al. entitled “Early cardiac dysfunction in pediatric patients on maintenance dialysis and post kidney transplant”, where the authors discussed the problem of “cardiorenal syndrome” which seems to be of high predictive importance in young patients with chronic kidney disease (CKD) on renal replacement therapy .The authors investigated heart changes, using imaging techniques for structural study and sensitive biomarkers to assess myocardial function and metabolism, in order to examine the association of these parameters with left ventricular hypertrophy (LVH). They calculated that cardiac magnetic resonance (CMR) and MR spectroscopy (MRS) allow the detection of cardiac dysfunction earlier. They also concluded that young patients with advanced CKD and normal ejection fraction demonstrated early cardiac impairments accompanied by increased LV mass index, which suggested a development of maladaptive hypertrophy.In this regard, we would like to pay attention to our observations made in children with CKD, which revealed the usefulness of a non-invasive 87-electrode electrocardiographic method, BSPM , in early detection of intraventricular conduction disturbances of various degrees.We have used isochrone maps that precisely reflect ventricular activation time (VAT maps). In the examined CKD children treated conservatively  or with Our results, being in strong relevance to the findings of Malatesta-Muncher et al., suggest that children with CKD, regardless of mechanisms responsible for LVH, adaptive or maladaptive, are at risk of developing progressive abnormalities in the intraventricular heart conduction system. Such disturbances of electrical activation propagation lead to delayed ventricular depolarization, which results in hemodynamic dyssynchrony and, consequently, with simultaneous left ventricular hypertrophy, worsen the global systolic function of the heart. Bearing this in mind, any possibilities enabling detection of subclinical cardiac dysfunction in CKD seem beneficial.


Determining the existence of interconnected responses among life-history traits and identifying underlying environmental drivers are recognized as key goals for understanding the basis of phenotypic variability. We studied potentially interconnected responses among senescence, fecundity, embryos size, weight of brooding females, size at maturity and sex ratio in a semiterrestrial amphipod affected by macroscale gradients in beach morphodynamics and salinity. To this end, multiple modelling processes based on generalized additive mixed models were used to deal with the spatio-temporal structure of the data obtained at 10 beaches during 22 months. Salinity was the only nexus among life-history traits, suggesting that this physiological stressor influences the energy balance of organisms. Different salinity scenarios determined shifts in the weight of brooding females and size at maturity, having consequences in the number and size of embryos which in turn affected sex determination and sex ratio at the population level. Our work highlights the importance of analysing field data to find the variables and potential mechanisms that define concerted responses among traits, therefore defining life-history strategies. Environment as an ecological actor causes variations in phenotype (phenotypic variability) and therefore in the performance of individuals  and the Atlantic Ocean, providing an ideal scenario to study how the environment affects life-history traits. This is particularly true in the talitrid amphipod iliensis , which iA. brasiliensis distribution, until at least two consecutive stations yielded no amphipods. At each station, a sheet metal cylinder (27 cm diameter) was used to remove the sediment up to a depth of 40 cm. Samples were field-sieved through a 0.5 mm mesh; samples with coarse sand were carefully sorted in the laboratory to retain all organisms. At each sampling station, sediment samples were collected with a corer (6.5 cm diameter) for determination of mean grain size, sand moisture and organic matter content. Measurements of sediment temperature and sand compaction were made, in the latter using a piston pocket penetrometer    could produce females, while smaller embryos  produce males  was described in several animal groups  EpiBulnheim ; and (3)Bulnheim . In thisBulnheim , the modce males .Atlantorchestoidea brasiliensis. Low-salinity values demand large amounts of energy for osmoregulation that cannot be assimilated as reserves (body weight) for reproductive costs. This explains why at low-salinity scenarios the individual weight of brooding females and the proportion of mature females were lower for a given size. In addition, salinity affected embryo size and fecundity, the latter through body weight. Low salinities represent a risk to offspring, being necessary to provide energy reserves (yolk) to each embryo to increase their probabilities to survive. Under this adverse scenario, the increase in embryo size and the decrease in female fecundity (due to the limited space in its pouch) represent a trade-off. Furthermore, yolk allocation could explain why large embryos produce females, determining the high prevalence of this sex at low salinities, which should compensate low individual fecundities. Our results highlighted that variables acting on physiology influence the energy balance and determine interconnected responses of life-history traits mainly associated with reproduction, having consequences at the population level. In addition, other variables related to environmental unpredictability (in our case beach slope) can have significant effects on fitness through organisms' survival. In summary, this study revealed that field data of multiple traits and modelling approaches that incorporate random factors are useful to unveil environmental drivers that define concerted responses of life-history traits, providing insights into understand the potential underlying mechanisms.Salinity was identified as the only variable that affects all life-history traits of


Further, enzymes at different nodes  of metabolic pathways influence enhancement of trehalose accumulation. Most of the study of trehalose biosynthesis was explored in medically significant Mycobacterium, research model Escherichia coli, industrially applicable Corynebacterium and food and probiotic interest Propionibacterium freudenreichii. Therefore, the present review dealt with the trehalose metabolism in these bacteria. In addition, an effort was made to recognize how enzymes at different nodes of metabolic pathway can influence trehalose accumulation.The current knowledge of trehalose biosynthesis under stress conditions is incomplete and needs further research. Since trehalose finds industrial and pharmaceutical applications, enhanced accumulation of trehalose in bacteria seems advantageous for commercial production. Moreover, physiological role of trehalose is a key to generate stress resistant bacteria by metabolic engineering. Although trehalose biosynthesis requires few metabolites and enzyme reactions, it appears to have a more complex metabolic regulation. Trehalose biosynthesis in bacteria is known through three pathways – OtsAB, TreYZ and TreS. The interconnections of  Escherichia coli, Corynebacterium sp. and Propionibacterium freudenreichii were reported previously , cold shock (4°C), heat shock (45°C) and against dehydration . Therefore, study of trehalose production and metabolism seems worth with this bacterium. The defence strategy of Corynebacterium glutamicum in osmotic stress was reported in the form of trehalose accumulation and three major pathways were involved with trehalose metabolism – OtsAB, TreYZ and TreS  in contrast to in vitro conditions  in Corynebacterium led to twofold increase in trehalose titre  resulted in sixfold increase in trehalose content . It has been extensively studied as it interconnects in vivo synthesis of glycogen and trehalose  were involved in synthesis of glycogen when trehalose was present in higher concentration   was reported in Propionibacterium shermanii , disaccharides (sucrose), polysaccharides (starch) and gluconeogenic (glycerol) were used for trehalose accumulation. Besides, the carbon channelizes through three important nodes of metabolic pathway – glucose-6-P, glucose-1-P and NDP-glucose as shown in Corynebacterium glutamicum indicated increased flux towards trehalose formation and it was principle osmolyte in low dilution rate  converts glucose-6-p to glucose-1-p which is used as substrate for synthesis of NDP-glucose by enzyme NDP-glucose pyrophosphorylase. In yeast, role of phosphoglucomutase for trehalose biosynthesis depends on various other factors like availability of glucose-6-phosphate Voit, . No direCorynebacterium and yeast. In fact, heterologous expression of galU genes together with otsAB from E. coli in Corynebacterium increased trehalose yield as discussed in above section  for enzyme OtsA. The influence of UDP-glucose pyrophosphorylase  was studied in et al., et al., Mycobacterium  , has important role in trehalose metabolism. This pathway was reported in the complete genome sequence of The solicitous study of trehalose metabolism is important in bacteria, as these can efficiently convert wastes from environment and can make trehalose production economical. It can be concluded that no particular enzyme has total control on trehalose biosynthesis, and similar conclusion was also predicted in yeast where optimum ratios of particular enzymes were found to have influence on trehalose biosynthesis as proposed by Jung and Stephanopoulos . It was


The versatility of the papers accepted to the topic is surprisingly broad. Three trends became evident, presumably reflecting the most vivid interests in the field: (1) the “Brain slice studies have played an important role in gaining an understanding of fundamental neuronal, synaptic, and network traits but this preparation has never been free from serious limitations. Among them repeatedly mentioned are: absence of blood flow, diffusion peculiarities, inadequate oxygenation/metabolic support, and neuronal damage due to the slicing procedure  that increases chances of ketogenesis (while decreasing glycolytic flux) is shown to have favorable effects on metabolic profiles, thus exerting neuroprotective properties (Zilberter, Together, the articles comprising this Research Topic depict a complex intricately organized system with representative studies at both the macro- and micro-levels (e.g., involving molecular signaling pathways; Venkateswaran et al.,


The interface between engineering and molecular life sciences has been fertile ground for advancing our understanding of complex biological systems. Engineered microstructures offer a diverse toolbox for cellular and molecular biologists to direct the placement of cells and small organisms, and to recreate biological functions in vitro: cells can be positioned and connected in a designed fashion, and connectivity and community effects of cells studied. Because of the highly polar morphology and finely compartmentalized functions of neurons, microfabricated cell culture systems and related on-chip technologies have become an important enabling platform for studying development, function and degeneration of the nervous system at the molecular and cellular level. Here we review some of the compartmentalization techniques developed so far to highlight how high-precision control of neuronal connectivity allows new approaches for studying axonal and synaptic biology. Over the past decades, molecular and cellular neuroscience has taken major steps towards understanding how the nervous system develops, functions and occasionally fails in disease. With billions of neurons interconnected via trillions of synapses, the complexity of mammalian brain poses a tremendous challenge for experimental scientists. Even reduced preparations, such as cultures of dissociated primary neurons, pose significant experimental challenges due to difficulties in controlling the growth and connectivity of axons and dendrites. Chemical cues and micropatterned surfaces have long been used to control growth of neurites in cell culture conditions. Neurons adhere and grow on glass surface coated with adhesive proteins but not on native glass. Finely defined patterns of adhesive proteins can be produced by selective coating and denaturation of neurite growth-promoting proteins  or by miAnother approach to control growth and connectivity neuronal processes is compartmentalized culture systems . The CamThe most commonly used way to achieve micro-scale compartmentalization is through the use of microfluidic chips. Microfluidic chips contain accurately patterned features in the range of 0.1 μm - 1 mm, and thus offer highly precise spatial and temporal control of cellular microenvironment. The traditional microfabrication materials are silicon and glass, but the microfluidics community has increasingly adopted polymers such as poly(dimethylsiloxane) (PDMS) as the materials of choice . TypicalOther approaches to study how cells respond to the topography of the substratum , have used silicon wafers and glass slides with holes and metal decorations -12. BondA commonly used approach is to utilize PDMS microfluidic chips sealed on top of glass. The wide adoption of PDMS stems mainly from ease of fabricating even large batches of chips through casting. The casting process (other than the master fabrication) also does not require any dedicated equipment, and can be performed anywhere. In addition, PDMS is a biocompatible and optically transparent polymer that forms reversible watertight seals when placed on smooth surfaces due to its elasticity . PDMS caSignificant advances have been made in developing analytical methods to monitor neuronal activity on a single cell level  -20. To mAgarose microstructures can be photothermally etched by area-specific melting of agar microchambers by spot heating using a focused laser beam of 1480 nm (which is absorbed by water and agar gel), and of a thin layer made of a light-absorbing material such as chromium with a laser beam of 1064 nm (since water and agar itself have little absorbance at 1064 nm) . For phaBy offering many significant methodological advantages, microfluidic techniques are reshaping the way axonal cell biology is studied in vitro. As compartmentalized microfluidic devices are based on geometric guidance of axon growth via engineered microgrooves, they provide a high level of control of connectivity and architecture of the neuronal network. This provides significant benefits for biochemical (e.g. isolation of axon-specific mRNA and protein species), cell biological  and electrophysiological studies  ,16,25-27The ability to image and quantify growth properties of axons and dendrites  in microfluidic devices is superior over more traditional methods ,28. MoreThe functionality of the axon and the presynaptic terminal is critically dependent on axonal trafficking of materials  and organelles such as mitochondria. Moreover, retrograde transport of target-derived signals, particularly neurotrophic proteins, is essential for neuronal survival and plasticity . Recent One of the main advantages of spatial segregation of axons is the possibility to investigate specific mechanisms of CNS axonal injury and degeneration, as well as possible regeneration . Since aBecause of their central role in biological computation and storage of information, synapses are of special interest in neurobiology. A synapse represents a very compact subcellular compartment with diameters less than 1 μm. Understanding the detailed molecular events that regulate presynaptic function requires high-resolution methods that provide quantitative information combined with molecular specificity. In recent years, numerous technological advances, including optogenetic methods and live-cell imaging, have facilitated probing the physiology of individual synapses. Combination of these techniques with microfluidic culture platforms offers a powerful way to study synaptic biology.Fluorescent organic dyes are widely used as reporters of dynamic changes in various cellular processes. Because calcium ions control key functions in all types of neurons, imaging dynamic local changes in intracellular calcium concentration has become an important experimental readout in neurobiology . PioneerElectrophysiological methods and in particular, the current clamp and voltage clamp techniques, provide the best temporal resolution for functional analysis of excitable cells. Intracellular recording methods typically target the postsynaptic cell, and allow high precision analysis of somatodendritic function. PDMS-based microfluidic technologies have been developed for high-throughput electrophysiological studies ,53. In tApart from the high-throughput systems, microfluidic chambers allowing compartment-specific manipulation provide valuable tools to study neuronal and in particular, axonal function. Geometrical limitations of PDMS-based culture platforms have made it challenging to combine conventional microelectrode-based patch clamp recording with microfluidic control of neuronal connectivity. Recently, a simple modular, three-layer PDMS chamber was developed for controlling neuronal connectivity and asymmetric viral transduction while also allowing whole cell patch clamp recording . TogetheNon-invasive extracellular measurement using a multi-electrode array (MEA) has been widely used in neuroscience for the past two decades -61, and Planar electrodes are suitable for monitoring fluctuation in the extracellular field potential produced by small groups of neurons when it is difficult to record and stimulate single neurons. In addition to the microwell-based approach of physically restraining connectivity of neurons with planar electrodes ,65, an iIn neuroscience, one of the main interests is how epigenetic information is processed and recorded as plasticity within a network pattern, and how changes in the network patterns relate to plasticity. Thus, for many years, neurobiologists have investigated single-cell-based neuronal network cultivation and examined the firing patterns of single neurons through the fabrication of cultivation substrates using microprinting techniques -71, pattUsing the photo-thermal etching method, microstructures can be formed within the agar layer on the chip by melting a portion of the agar layer at the spot of a focused infrared laser beam as shown in Figure An MEA system with an agar microchamber to control the topography of network patterns of neuronal networks in single cell level has been accomplished Figure C 74]. T. T74]. TAsymmetric microchannels in PDMS chips provide another way for creating unidirectional axon connectivity. Peyrin et al. recently described reconstruction of oriented cortico-striatal neuronal networks in “axon diode” microfluidic chambers composed of funnel-shaped microchannels . The axoHandling, sorting and analysis of small model organisms widely used in biology, Caenorhabditis elegans and Drosophila melanogaster, have been facilitated by developing special microfluidic devices . Preciseneurofluidics.Microfluidics has the potential to revolutionize cell biology. In particular, neurobiologists have found microfluidics capable of providing solutions to many methodological challenges. The number of device designs and applications is increasing rapidly, and microfluidic techniques are likely to be adopted widely by the neuroscience community in the near future. Integrated designs using combination of microfluidic chips with MEA, optogenetics and various types of biosensors  is expected to take these emerging on-chip technologies to new heights. Moreover, microfluidic devices are increasingly used with brain slices and could also provide great opportunities as scaffolds to develop implantable neural interfaces. Finally, utilization of the third growth dimension will provide another future direction to this new field of Aβ: β-amyloid peptide; BDNF: Brain-derived neurotrophic factor; CNS: Central nervous system; GFP: Green fluorescent protein; MEA: Multielectrode array; NGF: Nerve growth factor; PDMS: Poly(dimethylsiloxane); PLL: Poly-L-lysine.H.J.H is a cofounder and a shareholder of Hermo Pharma Ltd. Other authors declare no conflict of interest.CAB and PS performed literature review and contributed in writing the first draft of the manuscript. VJ and SF wrote the section on PDMS-related materials and manufacturing techniques. HT, FN, TK and KY wrote the sections on agar-microstructure technology and directed neuronal networks. SEL and CR performed literature review and contributed in writing the electrophysiological parts of the review. KY and HJH designed, edited and revised the final manuscript. All authors read and approved the final manuscript.


Shifting the role of counseling to less skilled workers may improve efficiency and coverage of health services, but evidence is needed on the impact of substitution on quality of care. This research explored the influence of delegating maternal and newborn counseling responsibilities to clinic-based lay nurse aides on the quality of counseling provided as part of a task shifting initiative to expand their role.Nurse-midwives and lay nurse aides in seven public maternities were trained to use job aids to improve counseling in maternal and newborn care. Quality of counseling and maternal knowledge were assessed using direct observation of antenatal consultations and patient exit interviews. Both provider types were interviewed to examine perceptions regarding the task shift. To compare provider performance levels, non-inferiority analyses were conducted where non-inferiority was demonstrated if the lower confidence limit of the performance difference did not exceed a margin of 10 percentage points.Mean percent of recommended messages provided by lay nurse aides was non-inferior to counseling by nurse-midwives in adjusted analyses for birth preparedness , danger sign recognition , and clean delivery . Lay nurse aides demonstrated superior performance for communication on general prenatal care , although non-inferiority was not achieved for newborn care counseling . The proportion of women with correct knowledge was significantly higher among those counseled by lay nurse aides as compared to nurse-midwives in general prenatal care , birth preparedness , and danger sign recognition . Both cadres had positive opinions regarding task shifting, although several preferred 'task sharing' over full delegation.Lay nurse aides can provide effective antenatal counseling in maternal and newborn care in facility-based settings, provided they receive adequate training and support. Efforts are needed to improve management of human resources to ensure that effective mechanisms for regulating and financing task shifting are sustained. Task shifting refers to the delegation of non-technical tasks traditionally held by professional workers to workers with lower qualifications . Recent Although the term is relatively new in the global health context, task shifting has been used for many years and in several countries. Numerous studies provide evidence across health-related areas on task shifting between non-physician clinicians and nurses or midwives -7. HowevHowever, given the magnitude of current health crises, addressing human resource needs through task shifting remains neglected , and theTask shifting has been praised on several fronts given its potential to improve the skill mix of teams ,22, to lThe World Health Organization (WHO) recently released guidelines on task shifting which emphasize the need to ensure that approaches are adopted as part of a broader strategy of health systems strengthening that includes mechanisms and research to make certain quality of care is not compromised ,31. To aThis study used a non-inferiority quasi-experimental design. A non-inferiority study was used because the quality of communication by lay nurse aides was not required to be significantly better than that of nurse-midwives, but it was necessary that the quality of communication be at least to the same standard . Seven pAntenatal care is traditionally conducted by trained nurse-midwives who provide antenatal clinical and communication services. Nurses and midwives in Benin undergo three years of standardized, government-accredited training in obstetric-gynecology and internal medicine. They are employees of the government with an average monthly salary of approximately $118 USD. Traditional roles of nurse-midwives consist of vital signs measurement, counseling, physical examination, and medication dispensation in antenatal care as well as management of deliveries and obstetrical complications. Lay nurse aides are also government employees with an average monthly salary of approximately $59 USD. They receive no formal education but rather are trained on-site by nurse-midwives to assist in tasks such as directing patient flow, taking vital signs, recording height and weight, record keeping, and cleaning. In the absence of a skilled provider, lay nurse aides may also informally provide other clinical services, although they are not trained to do so. To this extent, responsibilities sometimes overlap. The number of lay nurse aides often approximates that of nurse-midwives, although health facilities may have shortages of both cadres.We tested the influence of shifting the role of counseling to lay nurse aides on quality of counseling and maternal knowledge. Data were collected two weeks prior to the counseling task shift among women counseled by nurse-midwives (standard group) and following the task shift among women counseled by lay nurse aides (comparison group).A non-inferiority design examines whether a comparison is no worse or 'non-inferior' to a standard group. Because exact equivalence cannot be determined ,36, a maSeven public health centers with sufficient patient loads were selected to achieve the target sample size and increase the generalizability of findings among health centers with slightly varying service and patient characteristics. These facilities were operated by the Benin Ministry of Health and purposively selected within the Zou/Collines region among sites supported by the Integrated Health Project  under the management of University Research Co., LLC (URC). Participant eligibility criteria included being pregnant and presenting at the health facility for antenatal consultation during the data collection period, willingness to be interviewed after the clinic visit, and (when applicable) willingness to be counseled by a lay nurse aide with training identical to that of nurse-midwives in use of the counseling materials. Using systematic sampling, eligible women were approached while waiting for consultation, given information regarding the purpose of the study, and invited to participate. Participation from site managers and providers was obtained prior to the start of the study.This study was part of a larger study to evaluate the effectiveness of using a set of pictorial counseling cards to improve quality of antenatal counseling among nurse-midwives. Previous data collection in participating sites had shown that baseline levels of communication were poor and that women were not fully benefiting from health counseling by nurse-midwives during antenatal visits. Nurse-midwives were trained to use the job aids to improve their performance. Lay nurse aides were also trained to use the job aids to ensure that they had comparable performance support when assessing the effectiveness of the task shifting approach. Eleven antenatal counseling cards were organized into three modules to prioritize messages and ensure that over the course of pregnancy women would have multiple exposures to key information. These modules emphasized core messages relating to care during pregnancy, birth preparedness, danger signs, clean delivery, and newborn care Figure .A key component of task shifting is to define the task to be delegated and the training and experience needed for the type of worker to whom the shift will occur . The stuTo prepare for the task shift, the two cadres were trained for three days separately using similar curricula. Training of both provider types included a description of the concept of task delegation, peer and group role-playing, capacity building in interpersonal communication, and emphasis on quality of care. The nurse-midwife training was conducted in French and included technical materials in French and additional instruction on planning, supervision, and evaluation. The lay nurse aide training was comparable, but provided at a slower pace and conducted in the local language, Fon, since most lay nurse aides were not proficient in written and spoken French. The courses ended with a joint session of both cadres to ensure positive intra-provider relations and confirm roles related to task shifting.Prior to data collection, all sites received a supervisory visit from one of the trainers or technical advisors. These visits included a review of the organization of counseling using the counseling job aids, observation of consultations with direct feedback, and discussions about difficulties implementing the job aids or the task shift.There were three measurement areas in the study: quality of counseling, provider perceptions of task delegation, and women's knowledge of maternal and newborn care.To evaluate quality of counseling, providers' content of communication and counseling technique were measured through direct observation using a pre-tested observation checklist. The checklist covered five topic areas: general prenatal care, birth preparedness, dangers signs, clean delivery, and newborn care. 'General prenatal care' included four messages relating to prevention and treatment of malaria , iron/folate supplementation, having at least four antenatal visits, and information on diet and nutrition. 'Birth preparedness' included seven messages regarding identifying a place to deliver, identifying a skilled attendant, securing a means of transport, putting money aside, planning for emergencies, planning with a family member, and identifying a blood donor. 'Danger signs' highlighted nine maternal symptoms that require care: vaginal bleeding, convulsions, fever, water loss, abdominal pains, severe headaches, blurred vision, swelling of limbs, and absence or diminished fetal movement. 'Clean delivery' consisted of two messages relating to provision of a clean, plastic cloth for delivery and clean, dry towels for the mother and newborn. Six messages related to 'newborn care': skin-to-skin contact, early and exclusive breastfeeding, delayed bathing, clean cord care, and thermal protection. For each item, a trained observer selected 'yes' or 'no' depending on whether the woman received information regarding that item during her antenatal visit. Provider communication techniques were scored similarly across six communication techniques: presenting the subject, posing questions to determine current knowledge, using visual aid(s), verifying understanding, motivating adoption of new behaviors, and asking the woman if she has questions.e.g., age, education, qualification, years working in health field, years working at health center) was also obtained.Provider perceptions on task delegation were obtained using semi-structured questionnaires during individual interviews. Following the task shift, health workers were asked whether they thought that similar approaches should be introduced in other sites, what were perceived advantages and disadvantages, and what were recommended strategies to improve task shifting. Respondents were also asked to indicate whether they agreed (indicate 'yes') or disagreed (indicate 'no') to 14 statements regarding the organization, acceptability, and effectiveness of task shifting. Responses were coded and analyzed by topic area and statement. Information on provider demographic characteristics  of 10%. Non-inferiority (NI) would be demonstrated if the lower confidence limit for the difference in mean percent of recommended messages between the two provider groups lay above -∆ NI = -10. This would mean that the null hypothesis (H0: ∆ >∆NI) is rejected in favor of the alternative hypothesis (HA: ∆ <∆NI). Any difference smaller than the lower bound would be unlikely in the population. It is important to note that the upper limit of the confidence interval (CI) is not interpreted since the study is a one-sided trial, and observed improvements are consistent with the inference of non-inferiority [NI), where the difference surpasses -10, the results are inconclusive or provide insufficient evidence to support non-inferiority (U). If the lower limit lies completely above zero, superiority is demonstrated (S).The absence of statistical significance cannot be interpreted as equivalence ,36. Thiseriority . If the One-sided confidence intervals of the mean percent of recommended messages provided by the two provider groups were calculated using STATA . Two sample t-tests were used to examine bivariate differences. Because data were clustered, the study employed three-level hierarchal modeling techniques to account for the inherent correlation of data given that pregnant women (level 1) were nested within providers (level 2) who were nested within sites (level 3). Random effects were modeled for provider- and site-level characteristics. Fixed effects were modeled for patient characteristics among variables that significantly varied between groups. This statistical technique is more suitable for clustered data than conventional regression analyses that underestimate standard errors by assuming observations from the same sites or providers are unrelated ,39. Rath'Intention to treat' analyses in which patients are compared according to the assigned study arm and 'per protocol' analyses where patients are compared according to the study arm they actually received were conducted concurrently as recommended for non-inferiority studies ,35. The p ≤ 0.05.Maternal knowledge was analyzed using two sample tests of proportions and similar multivariate hierarchal regression to adjust for nesting of patient observations within providers and sites. Chi-squared descriptive statistics were used to calculate overall agreement with the task-shifting statements. Data were double entered using EpiData (Version 3.2) with automatic checks for range. In all analyses, the level of significance was considered at This study received ethics approval by the Johns Hopkins Bloomberg School of Public Health Institutional Review Board, Baltimore, Maryland; the Research & Evaluation review group of the USAID Health Care Improvement Project at University Research Co., LLC (URC), Bethesda, Maryland; and the USAID Integrated Family Health Project at URC, Bohicon, Benin.The study included 48 health care providers: 21 nurse-midwives and 27 lay nurse aides at seven sites  as compared to the lay nurse aide group . Mean gestational age for first-time attendees was 3.9 and 3.8 months for nurse-midwives and lay nurse aides, respectively (p > 0.05). Of the observed consultations, the primary language used was similar, with 97% of counseling sessions conducted in Fon in both groups (p > 0.05). On average, there were 6.9 providers per site (nurse-midwives plus lay nurse aides) with lay nurse aides slightly out numbering nurse-midwives (ratio = 1.29) (data not shown). Approximately 58 pregnant women were observed at each site representing approximately 9.8 observed consultations per nurse-midwife and 7.5 per lay nurse aide.Table By topic area, no significant differences in content of communication were observed in adjusted analyses between nurse-midwives and nurse aides in the area of birth preparedness, danger sign recognition, clean delivery, or newborn care. Non-inferiority was demonstrated among nurse aides for information on danger signs , clean delivery , and birth preparedness , but there was not sufficient evidence to demonstrate non-inferiority for messages relating to newborn care . Nurse aides had significantly higher performance in the area of general prenatal care as compared to nurse-midwives  . In adjusted models, correlation of observations within providers and sites slightly tapered the observed unadjusted effect (not reported), although all gains remained significant. Patient characteristics did not significantly influence performance scores.An item analysis of key messages within each topic area showed considerable variability in the proportion of women who received any one message Table . For somMean performance was high for both provider types with regard to communication techniques at 95% and 98% among nurse-midwives and lay nurse aides, respectively  , birth preparedness , and recognition of danger signs  Table . There wWith regard to staff perceptions on the organization of task shifting, most indicated that lay nurse aides could effectively counsel pregnant women if appropriately trained and supervised (98%) and that counseling could be done by both types of providers (98%) Table . HoweverFor statements relating to impact and effectiveness, most health workers reported that task shifting relieved skilled workers to focus on more clinical activities (93%), improved provider relationships (86%), and was more effective than the prior organization of care (86%). More than half (53%) of lay nurse aides indicated that counseling by lay nurse aides is more effective than that of nurse-midwives, who were less likely to agree (25%). Reasons for this belief were that lay nurse aides were closer to the communities, had fewer linguistic barriers, and had their training to rely on. About half of nurse aides (46%) and nurse-midwives (47%) were also concerned that lay nurse aide-led counseling is less effective because lay nurse aides needed the support of skilled providers who were more experienced in counseling and communication.Perceptions relating to comfort and acceptability were generally positive with strong agreement that counseling provided by lay nurse aides was acceptable to women (95%). Reasons given were that women do not distinguish between the qualifications of nurse-midwives or lay nurse aides and consider all health staff to be capable of providing services. The support for lay nurse aide-led counseling being done with ease-of-mind by nurse-midwives (61%) or lay nurse aides (74%) was also high. Providers who said they were comfortable with the task shift indicated that it gave more time to nurse-midwives for clinical activities, encouraged working more efficiently in teams, and enabled communication to be provided at a high level. Those who were not comfortable with the shift raised questions about the need for counseling by lay nurse aides at instances when skilled providers had sufficient time.Among open-ended questions, reported advantages to task shifting were that task shifting improved the continuity of services since nurse-midwives were often more occupied or likely to be absent more than lay nurse aides and that the shift clarified the role of lay nurse aides Table . In partThe World Health Organization (WHO) recently released a set of recommendations for task shifting to guide programming and policy for HIV and AIDS or other health areas 17). Table . Table 7An important criterion for task shifting is that there is no compromise in quality of care provided . This stAlthough non-inferior in several communication areas, lay nurse aides likewise had significantly higher mean scores in the area of general prenatal care and communication techniques without significant increases in duration of antenatal consultations as compared to nurse-midwives. We hypothesize that increases in general prenatal care may reflect lay nurse aides' increased motivation and better compliance to counseling job aid instructions. Similar conclusions were drawn in other studies ,41. GeneCommunication by lay nurse aides was also associated with better patient outcomes as evidenced by maternal knowledge. Although content and techniques were similar for both cadres, maternal knowledge may have been higher due to lay nurse aides' better interpersonal skills . It is aThe generally positive opinions by both types of health workers likely contributed to the intervention's success. Prior to the shift, the study team worked to garner support among stakeholders and determine perceptions of the quality of counseling. Most providers addressed this issue by talking about training and support. Thus, the task shifting approach emphasized building the capacity of those to whom the task was delegated and those who would provide supervision. This was achieved through training, role-playing, use of job aids, on-site supervision, and field support, as a well as open dialogue between providers to ensure that their specific skills were esteemed. Recent task shifting guidelines attribute use of these steps to overall effectiveness and sustainability of task shifting approaches .Non-inferiority was not demonstrated in the area of newborn care, although no significant differences were observed in unadjusted and adjusted analyses. Communication regarding care of the newborn was provided only to women in advanced pregnancy. Therefore, the smaller sample size for this sub-analysis likely contributed to wider confidence intervals in the mean percent of recommended messages that limited the study's power to detect non-inferiority. As a result, wider confidence intervals may have contributed to the inconclusive results. Even so, at the item level, there were two messages for which lay nurse aides were found to be significantly inferior -- both in the area of birth preparedness. These were identifying a skilled attendant and planning for an emergency. Communication performance may have been lower for these two messages given lay nurse aides' reluctance to distinguish between qualifications of providers for delivery care (since some lay nurse aides manage deliveries in the absence of a skilled provider). In addition, the message regarding planning for emergencies had few related pictorials on the counseling job aid, which lay nurse aides tended to more rely upon.Strengthening the pictorial and textual content of the job aid and exploring perceptions among lay nurse aides regarding selected key messages may improve performance. Efforts are needed likewise to strengthen the confidence of some lay nurse aides. The three-day training focused on all topic areas within communication in maternal and newborn care. However, one alternative may be to use a phased, stepwise approach in which less-skilled workers assume communication tasks gradually for specific topics. This would allow lay workers to gain confidence and competencies in one communication area before assuming another. Even so, training itself should not be viewed as the only strategy necessary for effective task shifting. Strong supervision is also vital. Although the study did not systematically assess supervision by nurse-midwives, anecdotal evidence suggests that supervision was not standardized, consistent, or thorough -- perhaps likewise contributing to lower confidence of lay nurse aides. How best to strengthen these processes will be integral in the effectiveness of task shifting approaches introduced at scale. This will include examining consequences of giving new supervision responsibilities to more skilled staff .It is worth noting that when examining providers' perceptions, there was limited support of full delegation of the counseling role to nurse aides. Responses pointed toward 'task sharing' rather than a complete shift. This varied from the tested approach in which lay nurse aides provided counseling and nurse-midwives supervised and managed clinical care. Other studies as well have recognized that full delegation may not be possible with preference given to task inclusion approaches . In thisThe limitations of this study deserve mention. This study did not use an experimental non-inferiority design in that patients were not randomized to either counseling by a nurse-midwife or lay nurse aide. Therefore, to minimize biases in which women were selectively counseled by provider type, the study had two adjacent data collection periods where sessions led by nurse-midwives were examined (prior to shift) followed by examination of sessions led by lay nurse aides (after the shift). This yielded comparable characteristics in the cohort of women observed in each group. Other limitations include the potentially conservative non-inferiority margin, an educated guess in the minimum allowable difference that was not clinically significant and that was feasible in light of available resources. Further study that includes a larger sample size and smaller margin may be warranted. In addition, the study did not assess costs associated with increased supervision to support task shifting, and there was no assessment of the quality of communication provided over time or throughout the pregnancy term. All sites were purposively selected for study participation among PISAF-supported health centers, potentially influencing the generalizability of the study's results to non-supported sites. Session observers could also not be blinded to the type of provider, which may have introduced biases, although no evidence was found of this.These findings have three main policy implications. One relates to regulatory frameworks needed to support training, organization, and evaluation mechanisms needed for task shifting. In Benin, the role of lay nurse aides as defined by the Ministry of Health does not include health education and counseling. Widespread use of lay nurse aides to provide counseling to pregnant women would necessitate revising current policies that not only outline appropriate tasks for the lay cadre, but also define strategies to ensure that lay nurse aides are sufficiently trained, supervised, and evaluated when assuming these tasks ,31,37. TAnother implication relates to the estimated costs of task shifting. Although commonly cited, savings have more recently been viewed as an unlikely benefit of task shifting and an inadequate justification for its implementation . This reFinally, this study identified some non-monetary incentives as perceived by lay nurse aides due to the expansion in their role. Such incentives included recognition from superiors, an opportunity to be more involved in patient care, and satisfaction from an expansion of professional competencies through capacity building. Many lay nurse aides indicated that they had not received this previously. This highlights the role of incentives in the uptake of task shifting, but points likewise to policy considerations regarding payment mechanisms for less skilled cadre in sustaining task shifting approaches ,25.This research aimed to generate evidence on the quality of antenatal counseling in maternal and newborn care by lay nurse aides as compared to nurse-midwives who traditionally assume this role. Communication by lay nurse aides with appropriate training, supervision, and job aids was found to be non-inferior to that of nurse-midwives with significant gains in maternal knowledge following antenatal consultation. This evidence along with the positive perceptions among providers is encouraging, but policy decisions should address mechanisms to appropriately regulate and monitor task shifting approaches prior to formal introduction. This includes identifying what kind of 'shift' or 'sharing' is most appropriate within broader efforts to improve management of human resources.The authors declare that they have no competing interests.LJ conceived and designed the study, developed the data collection instruments, supervised data collection, performed the statistical analysis, and wrote the manuscript. ASY and JA participated in the testing and finalization of the data collection instruments, conducted job aids training, participated in data collection, coordinated field implementation, reviewed the study results, and made contributions to the manuscript. MA participated in the design of the job aids and associated training, reviewed study results, and made contributions to the manuscript. AT assisted in the conception of the study, coordinated local support for research activities, and contributed to the manuscript. All authors have seen and approved the final manuscript.


Mylpharyngodon piceus), grass carp (Ctenopharyngodon idella), bighead carp , silver carp , common carp (Cyprinus carpio), and goldfish (Carassius auratus). All six of these species are found in the Mississippi River basin and tracking their invasion has proven difficult, particularly where abundance is low. Knowledge of the location of the invasion front is valuable to natural resource managers because future ecological and economic damages can be most effectively prevented when populations are low. To test the accuracy of environmental DNA (eDNA) as an early indicator of species occurrence and relative abundance, we applied eDNA technology to the six non-native cyprinid species putatively present in a 2.6 river mile stretch of the Chicago  canal system that was subsequently treated with piscicide. The proportion of water samples yielding positive detections increased with relative abundance of the six species, as indicated by the number of carcasses recovered after poisoning. New markers for black carp, grass carp, and a common carp/goldfish are reported and details of the marker testing to ensure specificity are provided.In many North American rivers, populations of multiple species of non-native cyprinid fishes are present, including black carp ( Hypophthalmichthys nobilis) and silver carp (H. molitrix) in the Chicago area waterway linking the Mississippi River and Great Lakes basins (USA), and demonstrated that eDNA had a greater detection sensitivity than traditional netting and electrofishing sampling methods Molecular tools are being used for detection of rare species in aquatic ecosystems Mylpharyngodon piceus) and grass carp (Ctenopharyngodon idella). These four species, along with two other non-indigenous cyprinid fishes, goldfish (Carassius auratus) and common carp (Cyprinus carpio), have been introduced to many regions of the world Asian carps (or Asian river carps) refer to a group of large-bodied (maximum >1 m long adult size) fish species of the family Cyprinidae that are native to large rivers of eastern Asia. In their native habitat, Asian carps migrate upstream to spawn in large rivers Since we did not utilize any vertebrate animals in this experiment directly (we sampled water and did not house or manipulate any species directly), no IACUC or animal welfare protocol was required for this study.On May 20, 2010, a 2.6 river-mile stretch of the Little Calumet River covering 173 surface acres was treated by a consortium of state and federal agencies with the piscicide rotenone . This waCyprinus carpio) and goldfish (Carassius auratus), we designed a single primer pair that amplifies both species because of their genetic similarity and the common occurrence of hybridization in wild populations We used previously published species-specific Polymerase Chain Reaction (PCR) primers for bighead and silver carp detection http://ncbi.nlm.nih.gov) for each of the target species along with several native fishes (see below) considered either closely related or that might co-occur with the target species in the Mississippi River Basin. Both black carp and grass carp are the only known species within their respective genera , which reduces the likelihood that any native (or other non-native) cyprinids in North America might cross-amplify with the developed markers. For each species, sequence data obtained from GenBank were aligned using BioEdit in silico using the computer software package AlleleID  or the Primer-BLAST function in Genbank (http://www.ncbi.nlm.nih.gov/tools/primer-blast/).To develop primers, mitogenomic data were gathered from GenBank , largemouth bass , gizzard shad (Dorosoma cepedianum), and round goby . These closely related and non-closely related non-target species were representative of multiple major taxonomic groups in the region.Candidate primer sets were screened for positive amplification for their intended target species in the lab using genomic DNA extracts from tissues of the target organisms. Additionally, candidate primer sets were screened on multiple non-target species to ensure specificity of the markers to only intended target species. Targeted PCR amplification reactions were performed on genomic DNA extractions from multiple individuals of target species ). All candidate primer pairs were tested on genomic DNA from non-target species  including goldfish, common carp, black carp, grass carp, bighead carp, silver carp, fathead minnows , 2.5 mM Mg(OAc)2, 10 nmol each of dNTP, DNA template, species-specific primers , were not apparent in this study; otherwise we would have had positive eDNA detections of bighead and silver carp on the day of rotenone treatment when they were absent from the system. Previously bighead and silver carp had been detected in the treatment reach , and whiAlthough genetic markers presented here were designed for eDNA surveillance of waterways, similar markers could equally be used to identify cryptic species or to verify specific species identifications when morphological characteristics have been compromised . In addition, water samples from a variety of sources in addition to natural lentic and lotic habitats could be analyzed for eDNA to determine if target species are indeed present, including reservoirs, farm ponds, ships’ ballast tanks, bait tanks, aquarium imports, and tanks on trucks used to transport live fish.For example, the US Lacey Act list of injurious wildlife prohibits the importation and interstate transport of live black carp, bighead carp, and silver carp. In addition, most states regulate or restrict use of any existing stocks of black carp and grass carp, including (diploids and triploids) within their borders Finally, environmental DNA surveillance technology is rapidly evolving with refinements of methodology and advances in experimentation


We examined associations between neighborhood socioeconomic disadvantage, perceived neighborhood safety and cardiometabolic risk factors, adjusting for health behaviors and socioeconomic status (SES) among African Americans.Study participants were non-diabetic African Americans  in the baseline examination (2000–2004) of the Jackson Heart Study. We measured eight risk factors: the metabolic syndrome, its five components, insulin resistance and cardiovascular inflammation. We assessed neighborhood socioeconomic disadvantage with US Census 2000 data. We assessed perceived neighborhood safety, health behaviors and SES via survey. We used generalized estimating equations to estimate associations with a random intercept model for neighborhood effects.After adjustment for health behaviors and SES, neighborhood socioeconomic disadvantage was associated with the metabolic syndrome in women . Lack of perceived safety was associated with elevated glucose  and waist circumference  among women, and with elevated glucose  and insulin resistance  among men.Neighborhood socioeconomic disadvantage and perceived safety should be considered as targets for intervention to reduce cardiometabolic risks among African Americans. Cardiometabolic risk factors, including abdominal obesity, elevated blood pressure, dyslipidemia, glucose intolerance and inflammation are highly prevalent among African Americans independent associations with cardiometabolic risk factors, above and beyond associations with health behaviors, has not been fully explored in African Americans neighborhood socioeconomic disadvantages, including high unemployment and poverty, as well as high levels of perceived social disorder, such as lack of perceived neighborhood safety Increasingly, the residential neighborhood socioeconomic environments to which African Americans are exposed are hypothesized to contribute to cardiometabolic risks in these groups Recent population-based studies have shown independent associations between neighborhood disadvantage and cardiometabolic risks. The Atherosclerosis Risk in Communities study found that low neighborhood socioeconomic status (SES) was associated with an increased prevalence of the metabolic syndrome among African American women in the southeastern US, but not among African American men, a finding that was not completely explained by alcohol use, smoking, or physical activity among women To date, associations between neighborhood socioeconomic disadvantage, perceived disorder and specific cardiometabolic risk factors have yet to be fully explored among African Americans. The 2004 National Heart, Lung and Blood Institute/American Heart Disease Association conference on the definition of metabolic syndrome described specific etiologic cardiometabolic factors that may underlie the pathogenesis of the metabolic syndrome: (1) abdominal fat around the waist, (2) insulin resistance, and (3) other mechanisms such as cardiovascular inflammation Thus, our study examined cross-sectional associations between features of neighborhood socioeconomic disadvantage, health behaviors, and cardiometabolic risk factors among African American participants in the Jackson Heart Study (JHS) who reside in the Jackson, Mississippi Metropolitan Statistical Area (MSA), a geographic area with a high prevalence of cardiovascular risk factors. Specifically, we examined whether socioeconomic aspects of residential neighborhoods  are associated with health behaviors , cardiometabolic risk factors  and biomarkers associated with abdominal obesity, inflammation and insulin resistance. We hypothesized that neighborhood socioeconomic disadvantage and perceived neighborhood safety would have independent associations with the metabolic syndrome and its constituent components, above and beyond associations with health behaviors. We additionally hypothesized that bio-measures of three etiologic factors, abdominal adiposity, cardiovascular inflammation and insulin resistance were associated with neighborhood socioeconomic disadvantage and perceived neighborhood safety. We examined sex differences in the above associations.The JHS is an observational cohort study designed to investigate factors that influence the development and worsening of CVD in African Americans Methods describing participant recruitment and sample characteristics have been reported previously We examined eight measures of cardiometabolic risk: the metabolic syndrome, the five constituent components of the metabolic syndrome ; a measure of cardiovascular inflammation, c-reactive protein (CRP); and a measure of insulin resistance, as defined by the homeostasis model assessment of insulin resistance (HOMA-IR) Eight hour fasting triglyceride, HDL cholesterol, insulin and glucose levels were obtained via venipuncture. Assays and procedures for measuring fasting whole blood, plasma and serum-derived analytes in the JHS have been previously described socioeconomic disadvantage ; racial heterogeneity and family structure ; urbanicity (percentage of neighborhoods in urban areas) and residential stability . We used exploratory factor analysis with an oblique PROMAX rotation to construct an index of neighborhood socioeconomic disadvantage from these ten candidate measures. An a priori factor loading (standardized regression coefficient) of 0.50 was used to determine whether a covariate loaded on any extracted factor. The reliability of the neighborhood socioeconomic disadvantage index was assessed via standardized Cronbach’s alpha coefficients.We defined neighborhoods at the census tract level using the 2000 US Census (Census 2000) boundaries. We used the theory of Social Disorganization to select measures of neighborhood socioeconomic conditions, in order to create an index of neighborhood socioeconomic disadvantage in Jackson MSA neighborhoods JHS survey data assessed neighborhood safety perceptions among individual participants with a single item statement “this neighborhood is safe from crime.” Data on perceived safety were collected during an annual follow-up call conducted approximately on the third anniversary of their clinic visit date. Participants who agreed or strongly agreed were categorized as perceiving their neighborhood as safe; those who disagreed or strongly disagreed were categorized as perceiving their neighborhood as unsafe.health behaviors , the individual’s socioeconomic position , as well as the participant’s age and sex. We measured physical activity among JHS participants with a validated active living index, which is a summary score of the (1) frequency and duration of activities performed during leisure-time, and lifestyle activities including walking and biking during errands, and while commuting to and from work, (2) minus the frequency of television watching as an indicator of sedentary behavior We used self-reported measures of covariates related to cardiometabolic risk factors, and exposure to neighborhood socioeconomic disadvantage including Consistent with ATP III guidelines, we assessed cardiometabolic risk factors for the metabolic syndrome among non-diabetic patients. We excluded 1,236 (23%) participants with diabetes, defined as fasting plasma glucose ≥126 mg/dL, a self-reported physician’s diagnosis of diabetes, or self-reported use of anti-glycemic medications After the above exclusions, in multivariable models, we additionally excluded 51 individuals whose residential addresses could not be assigned to one of the 102 JHS neighborhoods (census tracts). We also excluded individuals living in census tracts with very few numbers of JHS participants per neighborhood (fewer than 10), resulting in the additional exclusion of 90 JHS participants and 21 (21%) Jackson MSA neighborhoods Descriptive statistics at the individual level were performed with Chi-squared tests of associations among proportions, and Kruskal-Wallis non-parametric tests for associations between categorical and continuous variables.To estimate associations between each cardiometabolic risk factor, neighborhood-level condition, and individual-level covariate, we estimated prevalence ratios (PR) using log binomial regression, and odds ratios (OR) using binomial regression with a logit link, within a generalized estimating equations framework in the PROC GENMOD procedure in SAS® neighborhood socioeconomic disadvantage: the percentage of adults living in poverty (factor loading 0.82), the civilian unemployment rate (factor loading 0.77), the percentage of households with no car ownership (factor loading 0.97), and the percentage of owner occupied housing in the neighborhood (factor loading −0.55). The constructed index, constructed as the sum of all four covariates weighted by their factor loadings, had poor reliability . Thus, the neighborhood socioeconomic disadvantage (NSED) index was re-constructed as the weighted sum of the three covariates with the highest factor loadings . The resultant index had excellent reliability .The exploratory factor analysis identified four covariates that “loaded” on, or were tightly associated with, the first extracted factor which we identified as neighborhood stability (percentage of neighbors in place since 1995) was not associated with neighborhood disadvantage ; this measure was included as a separate covariate in subsequent multivariable models.The NSED index ranged from a minimum value of 0.82 (low disadvantage) to a maximum of 97.8 (high disadvantage) with a median value of 18.1 and standard error of 2.0. There were no sex differences in living in the most advantaged and disadvantaged neighborhoods. There were sex differences in median family household income and perceived neighborhood safety. Compared to men, women had lower median household incomes , and were more likely to perceive their neighborhoods as unsafe  [data not shown in tables].lower prevalence of low HDL-C  compared to men in the most advantaged neighborhoods. Neighborhood socioeconomic disadvantage was not associated with either CRP or HOMA-IR in men or women. Perceived neighborhood safety was not associated with metabolic syndrome among women in any of the sequential multivariable models. Neither neighborhood socioeconomic disadvantage nor perceived neighborhood safety were associated with the metabolic syndrome in men .lower prevalence of insulin resistance among women. Among men, perceived neighborhood safety was associated with elevated HOMA-IR after adjustment for covariates , but not elevated waist circumference or inflammation. Neighborhood socioeconomic disadvantage was not associated with insulin resistance, elevated waist circumference or inflammation among men. Increased physical activity was associated with a lower prevalence of all three biomeasures among men , and where the lack of perceived safety was highly common. Additionally, at the time they were surveyed, JHS participants in this study were non-diabetic, but were at a stage in the life-course in which a vast majority already experienced metabolic derangements associated with aging and chronic stress, particularly abdominal adiposity and cardiovascular inflammation among women, and elevated blood pressure among men and women Our findings are concordant with work in randomized controlled trial settings among urban African American women that found moving from high to low poverty neighborhoods was associated with reductions in extreme obesity and prevalent diabetes Our study adds to prior work by contributing rarely available data on specific physiologic mechanisms associated with disadvantage and disorder, particularly, measures of insulin resistance and inflammation among African Americans. Though each of these pathways is associated with HPA-axis stimulation, our study strengthens the hypothesis that glucose metabolism, and insulin resistance may be more closely tied to neighborhood disadvantage and disorder than other features of HPA-axis stimulation, namely dyslipidemia, as also seen in the Neighborhood Deprivation and Cardiometabolic Risk Factors in the Diabetes Study of Northern California (DISTANCE) study by Laraia et al. Our study also adds to prior work by including a measure of neighborhood socioeconomic disadvantage with strong reliability in neighborhoods in the southeastern United States, which showed convergent validity with other concepts rooted in the literature on Social Disorganization Theory Our data have important limitations that should be considered. First, the study design was cross-sectional and we cannot provide information on the causal relation between neighborhood disadvantage, disorder, and cardiometabolic risks. Our measure of perceived neighborhood safety, in particular, was assessed retrospectively and may have underestimated the strength of these associations compared to a concurrent assessment of perceived safety. Moreover, there were missing data on perceived neighborhood safety due to the timing of survey administration, and on income due to item non-response. We used multiple imputation techniques to avoid bias associated with list-wise deletion of these responses In summary, our work contributes to the literature by finding specific cardiometabolic risks factors that are independently associated with neighborhood disadvantage and disorder in middle-aged and older African American populations. Glucose metabolism and insulin resistance should be targeted and tracked in intervention studies designed to mitigate and eliminate material and psychosocial disadvantages associated with neighborhood contexts. Importantly, among JHS participants, more than 70% were exposed to disadvantaged neighborhoods, and a substantial percentage perceived that these neighborhoods were unsafe. Given the pervasive nature of these exposures, population-level interventions against neighborhood disadvantage and disorder have great potential to impact the biology of disadvantage that we observed, and should be considered as a strategy to achieve health equity for African American populations in high risk places.Table S1Contextual Characteristics: Defining Neighborhood Socioeconomic Disadvantage in Jackson Heart Study Neighborhoods. Figures represent ecological associations at the neighborhood (census tract) level. Kruskal-Wallis statistic used to test for non-parametric associations between categorical neighborhood disadvantage and continuous measures of related contextual characteristics. Related contextual characteristics are measures from the Theory of Social Disorganization.(DOCX)Click here for additional data file.