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Social quarantine has severely impacted the training of new scientists, but the use of virtual platforms can supplement mentorship, making career development seamless.

Despite expectations that Medicare accountable care organizations (ACOs) would curb health care spending, their effect has been modest. One possible explanation is that ACOs' inability to prohibit out-of-network care limits their control over spending. To examine this possibility, we examined the association between out-of-network care and per beneficiary spending using national Medicare data for 2012-15. While there was no association between out-of-network specialty care and ACO spending, each percentage-point increase in receipt of out-of-network primary care was associated with an increase of $10.79 in quarterly total ACO spending per beneficiary. When we broke down total spending by place of service, we found that out-of-network primary care was associated with higher spending in outpatient, skilled nursing facility, and emergency department settings, but not inpatient settings. Our findings suggest an opportunity for the Medicare program to realize substantial savings, if policy makers developed explicit incentives for beneficiaries to seek more of their primary care within network.

Dose-response models (DRMs) are used to predict the probability of microbial infection when a person is exposed to a given number of pathogens. In this study, we propose a new DRM for Staphylococcus aureus (SA), which causes skin and soft-tissue infections. The current approach to SA dose-response is only partially mechanistic and assumes that individual bacteria do not interact with each other. Our proposed two-compartment (2C) model assumes that bacteria that have not adjusted to the host environment decay. After adjusting to the host, they exhibit logistic/cooperative growth, eventually causing disease. The transition between the adjusted and un-adjusted states is a stochastic process, which the 2C DRM explicitly models to predict response probabilities. By fitting the 2C model to SA pathogenesis data, we show that cooperation between individual SA bacteria is sufficient (and, within the scope of the 2C model, necessary) to characterize the dose-response. This is a departure from the classical single-hit theory of dose-response, where complete independence is assumed between individual pathogens. From a quantitative microbial risk assessment standpoint, the mechanistic basis of the 2C DRM enables transparent modeling of dose-response of antibiotic-resistant SA that has not been possible before. It also enables the modeling of scenarios having multiple/non-instantaneous exposures, with minimal assumptions.

Carter et al. report that exposure to static magnetic and electric fields (sBE), for as little as 3 days, reverses glucose intolerance and insulin resistance in diet-induced and genetic mouse models of type 2 diabetes (1). They hypothesize that sBE triggers a systemic redox response to modulate insulin sensitivity and that sBE could therefore be used as a noninvasive treatment for type 2 diabetes. However, these authors were unable to define a mechanism to explain how SBE might alter ROS or to identify the specific proteins that mediate this effect. Given these limitations we propose an alternative hypothesis to explain their findings.

Primary cilia are microtubule-based organelles present on most cells that regulate many physiological processes, ranging from maintaining energy homeostasis to renal function. However, the role of these structures in the regulation of behavior remains unknown. To study the role of cilia in behavior, we employ mouse models of the human ciliopathy, Bardet-Biedl Syndrome (BBS). Here, we demonstrate that BBS mice have significant impairments in context fear conditioning, a form of associative learning. Moreover, we show that postnatal deletion of BBS gene function, as well as congenital deletion, specifically in the forebrain, impairs context fear conditioning. Analyses indicated that these behavioral impairments are not the result of impaired hippocampal long-term potentiation. However, our results indicate that these behavioral impairments are the result of impaired hippocampal neurogenesis. Two-week treatment with lithium chloride partially restores the proliferation of hippocampal neurons which leads to a rescue of context fear conditioning. Overall, our results identify a novel role of cilia genes in hippocampal neurogenesis and long-term context fear conditioning., Author summary The primary cilium is a microtubule-based membranous projection on the cell that is involved in multiple physiological functions. Patients who have cilia dysfunction commonly have intellectual disability. However, it is not known how cilia affect learning and memory. Studying mouse models of a cilia-based intellectual disability can provide insight into learning and memory. One such cilia-based intellectual disability is Bardet-Biedl Syndrome (BBS), which is caused by homozygous and compound heterozygous mutations of BBS genes. We found that a mouse model of BBS (Bbs1M390R/M390R mice) has learning and memory defects. In addition, we found that other mouse models of BBS have similar learning and memory defects. These BBS mouse models have difficulty associating an environment with a painful stimulus, a task designed to test context fear memory. This type of memory involves the brain hippocampus. This brain region produces new cells even into adulthood. The rate of new cell production in the hippocampus is important for learning and memory. Bbs1M390R/M390R mice have decreased cell production in the hippocampus. Treating Bbs1M390R/M390R mice with a compound (lithium) that increases cell production in the hippocampus improved the learning and memory deficits. Our results demonstrate a potential role for cilia in learning and memory, and indicate that lithium is a potential treatment, requiring further study, for the intellectual disability phenotype of BBS.

Among numerous challenges facing space travellers, microbial infection is one of the unknown risks associated with human spaceflight. Prevention and control of microbial infections are of critical concern during space missions. The objective of this research is to develop a quantitative microbial risk assessment (QMRA) to model the risk of Staphylococcus aureus skin infection and mitigation strategies that may effectively reduce skin infection risks. QMRA was carried out by incorporating the level of S. aureus contamination from International Space Station Microbial Observatory Experiment, bacterial transfer rate to skin, growth pattern of S. aureus on skin, space travellers’ daily behaviour and dose-response model. The results demonstrate that a daily skin cleaning regimen has a significant effect on reducing the skin infection risks. Once a day skin cleaning reduces infection risk by 84.2% and twice a day skin cleaning can reduce the risk of skin infection by 96.1% during a seven-day space mission. Frequency of contact with contaminated surfaces and time elapsed between cleaning events are the most important input parameters that contribute to the overall risk outcome. There are degrees of uncertainties associated with the predicted outcomes when interpreted by itself due to the limitation of microbial data and the dose-response model that derived from a short-term clinical study on Earth. The comparative risk analysis as used in this study offers a scientific basis regarding the effectiveness of interventions (skin cleaning regimens) in mitigating skin infection risks during spaceflight.

In September 2017, two category-5 hurricanes Irma and Maria swept through the Caribbean Sea in what is now known as the region’s most active hurricane season on record, leaving disastrous effects on infrastructure and people’s lives. In the U.S. Virgin Islands, rain cisterns are commonly used for harvesting roof-top rainwater for household water needs. High prevalence of Legionella spp. was found in the cistern water after the hurricanes. This study carried out a quantitative microbial risk assessment to estimate the health risks associated with Legionella through inhalation of aerosols from showering using water from cisterns after the hurricanes. Legionella concentrations were modeled based on the Legionella detected in post-hurricane water samples and reported total viable heterotrophic bacterial counts in cistern water. The inhalation dose was modeled using a Monte Carlo simulation of shower water aerosol concentrations according to shower water temperature, shower duration, inhalation rates, and shower flow rates. The risk of infection was calculated based on a previously established dose–response model from Legionella infection of guinea pigs. The results indicated median daily risk of 2.5 × 10−6 to 2.5 × 10−4 depending on shower temperature, and median annual risk of 9.1 × 10−4 to 1.4 × 10−2. Results were discussed and compared with household survey results for a better understanding of local perceived risk versus objective risk surrounding local water supplies.

Scientific success is mainly supported by mentoring, which often occurs through face-to-face interactions. Changes to the research environment incurred by the Coronavirus 2019 (COVID-19) pandemic have necessitated mentorship adaptations. Here, we describe how mentors can broaden their mentorship to support trainee growth and provide reassurance about trainee development amid uncertain circumstances.

Summary Aberrant redox signaling underlies the pathophysiology of many chronic metabolic diseases, including type 2 diabetes (T2D). Methodologies aimed at rebalancing systemic redox homeostasis have had limited success. A noninvasive, sustained approach would enable the long-term control of redox signaling for the treatment of T2D. We report that static magnetic and electric fields (sBE) noninvasively modulate the systemic GSH-to-GSSG redox couple to promote a healthier systemic redox environment that is reducing. Strikingly, when applied to mouse models of T2D, sBE rapidly ameliorates insulin resistance and glucose intolerance in as few as 3 days with no observed adverse effects. Scavenging paramagnetic byproducts of oxygen metabolism with SOD2 in hepatic mitochondria fully abolishes these insulin sensitizing effects, demonstrating that mitochondrial superoxide mediates induction of these therapeutic changes. Our findings introduce a remarkable redox-modulating phenomenon that exploits endogenous electromagneto-receptive mechanisms for the noninvasive treatment of T2D, and potentially other redox-related diseases.

Evidence linking electronic health record (EHR) adoption to better care is mixed. More nuanced measures of adoption, particularly those that capture the common incremental approach of adding functions over time in US hospitals, could help elucidate the relationship between adoption and outcomes. We used data for the period 2008-13 to assess the relationship between EHR adoption and thirty-day mortality rates. We found that baseline adoption was associated with a 0.11-percentage-point higher rate per function. Over time, maturation of the baseline functions was associated with a 0.09-percentage-point reduction in mortality rate per year per function. Each new function adopted in the study period was associated with a 0.21-percentage-point reduction in mortality rate per year per function. We observed effect modification based on size and teaching status, with small and nonteaching hospitals realizing greater gains. These findings suggest that national investment in hospital EHRs should yield improvements in mortality rates, but achieving them will take time.

Isoproterenol (ISO), is a non-selective beta-adrenergic agonist, that is used widely to induce cardiac injury in mice. While the acute model mimics stress-induced cardiomyopathy, the chronic model, administered through an osmotic pump, mimics advanced heart failure in humans. The purpose of the described protocol is to create the chronic ISO-induced heart failure model in mice using an implanted mini-pump. This protocol has been used to induce heart failure in 100+ strains of inbred mice. Techniques on surgical pump implantation are described in detail and may be relevant to anyone interested in creating a heart failure model in mice. In addition, the weekly cardiac remodeling changes based on echocardiographic parameters for each strain and expected time to model development are presented. In summary, the method is simple and reproducible. Continuous ISO administered via the implanted mini-pump over 3 to 4 weeks is sufficient to induce cardiac remodeling. Finally, the success for ISO model creation may be assessed in vivo by serial echocardiography demonstrating hypertrophy, ventricular dilation, and dysfunction.

Background Health Information Organizations (HIOs) are third party organizations that facilitate electronic health information exchange (HIE) between providers in a geographic area. Despite benefits from HIE, HIOs have struggled to form and subsequently gain broad provider participation. We sought to assess whether market-level hospital and EHR vendor dynamics are associated with presence and level of hospital participation in HIOs. Methods 2014 data on 4523 hospitals and their EHR vendors were aggregated to the market level. We used multivariate OLS regression to analyze the relationship between hospital and vendor dynamics and (1) probability of HIO presence and (2) percent of hospitals participating in an HIO. Results 298 of 469 markets (64%) had HIO presence, and in those markets, 47% of hospitals participated in an HIO on average. In multivariate analysis, four characteristics were associated with HIO presence. Markets with more hospitals, markets with more EHR vendors, and markets with an EHR vendor-led HIE approach were more likely to have an HIO. Compared to markets with low hospital competition, markets with high hospital competition had a 25 percentage point lower probability of HIO presence. Two characteristics were associated with level of hospital HIO participation. Markets with more hospitals as well as markets with high vendor competition (compared to low competition) had lower participation. Conclusion Both hospital and EHR vendor dynamics are associated with whether a market has an HIO as well as the level of hospital participation in HIOs. Electronic supplementary material The online version of this article (10.1186/s12911-018-0605-y) contains supplementary material, which is available to authorized users.

Objective To assess whether the level of health information exchange (HIE) in U.S. hospitals is related to technology capabilities, incentives to exchange, or both. Study Setting A total of 1,812 hospitals attesting to stage 2 of Medicare's Meaningful Use Incentive Program through April 2016. Study Design Hospital-level, multivariate OLS regression with state fixed effects was used to analyze the relationship between technology capability and incentives measures, and percent of care transitions with summary of care records (SCRs) sent electronically to subsequent providers. Principal Findings Stage 2 hospitals reported sending SCRs electronically for an average of 41 percent (median = 33 percent) of transitions. HIE level is related to four capability measures, one incentive measure, and one measure that is related to both capability and incentive. Percent of transitions with SCRs sent electronically was 3 percentage points higher (95 percent CI: 0.1–5.1) for hospitals with a third-party HIE vendor, 3 percentage points higher (95 percent CI: 0.5–5.4) for hospitals with an EHR vendor as their HIE vendor, and 3 percentage points higher (95 percent CI: 0.4–5.4) for hospitals that automatically alert primary care providers. The direction and statistical significance of the relationships between specific EHR vendor and electronic SCR transmission level varied by vendor. Nonprofits and government hospitals performed 5 percentage points higher (95 percent CI: 1.5–9.1) and 8 percentage points higher (95 percent CI: 3.4–12.3) than for-profits. Hospitals in systems performed 3 percentage points higher (95 percent CI: 0.8–6.1). Conclusion The overall level of HIE is low, with hospitals sending an SCR electronically for less than half of patient transitions. Specific hospital characteristics related to both technology capabilities and incentives were associated with higher levels of HIE.

Carbon nanomaterial coatings have been widely investigated for many biomedical applications including bone tissue engineering. Current methods to fabricate carbon nanomaterial coatings are limited by specific substrate requirements and the lack of strong bonds between the nanomaterials. Furthermore, few studies compare the effect of carbon nanoparticle architecture on stem cell differentiation and mineralization for osteogenic differentiation. Herein, we report a study comparing chemically crosslinked carbon nanotubes (of various diameters), graphene nanoplatelets, and graphene nanoribbons coatings for adipose derived stem cell differentiation toward an osteogenic lineage. We observed greatest autodeposition of calcium on graphene nanoribbon surfaces, while multiwalled carbon nanotubes of high diameter had the greatest influence on stem cell fate (by alkaline phosphatase activity, calcium deposition, and osteocalcin measurements). Studies indicate the cause for multiwalled carbon nanotube related stem cell differentiation, may be related to early timepoint toxicity as indicated by lactose dehydrogenase release. These results indicate suggestions for orthopedic tissue engineering applications for carbon nanomaterial coatings. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1189-1199, 2018.

These data represent the abundance, diversity and predicted function gene profiles of the microbial communities present on human skin before and after swimming in the ocean. The skin microbiome has been shown to provide protection against infection from pathogenic bacteria. It is well-known that exposure to ocean water can cause skin infection, but little is known about how exposure can alter the bacterial communities on the skin. Skin microbiome samples were collected from human participants before and after swimming in the ocean. These data were used to analyze the changes in abundance and diversity of microbial communities on the skin and the changes in the functional profiles of the bacteria, specifically focusing on genes involved in antibiotic resistance and bacterial virulence.

The skin is a complex organ responsible for protecting the body from physical, chemical and biological insults. The skin microbiome is known to play an important role in protecting the host from skin infections. This study examined the skin microbiome and the changes in antibiotic resistance genes (ARGs), antibiotic biosynthesis genes (ABSGs) and virulence factor genes (VFGs) on human skin before and after swimming in the ocean. Skin microbiome samples were collected from human participants before and after they swam in the ocean, and at 6 h and 24 h post-swim. The samples were analyzed using 16S rRNA gene and shotgun metagenomic sequencing. The results showed that not only is the skin microbiome composition altered after swimming, but the abundance and diversity of ARGs, ABSGs and VFGs on the skin increased post-swim. Overall, there was an increase in total ARGs by 70.6% from before to after swimming. The elevated number of ARGs persisted and continued to increase for at least 6 h post-swim with greater than a 300% increase in comparison with samples collected before ocean swimming. The outcomes of the study support the epidemiological observations of increased risk of skin infections after swimming in the ocean. Cleaning the skin immediately after recreational ocean activities is recommended to reduce the opportunity for infection.

Genetically encoded fluorescent protein tags revolutionized proteome studies, while the lack of intrinsically fluorescent RNAs has hindered transcriptome exploration. Among several RNA-fluorophore complexes that potentially address this problem, RNA Mango has an exceptionally high affinity for its thiazole orange (TO)-derived fluorophore, TO1-Biotin (Kd ~3 nM), and in complex with related ligands, is one of the most red-shifted fluorescent macromolecular tags known. To elucidate how this small aptamer exhibits such properties, which make it well suited for studying low-copy cellular RNAs, we determined its 1.7 Å resolution co-crystal structure. Unexpectedly, the entire ligand, including TO, biotin, and the linker connecting them, abuts one of the near-planar faces of the three-tiered G-quadruplex. The two heterocycles of TO are held in place by two loop adenines and make a 45° angle with respect to each other. Minimizing this angle would increase quantum yield and further improve this tool for in vivo RNA visualization., Graphical Abstract

De facto reuse refers to the use of surface water that contains a considerable portion of wastewater effluent from upstream communities as a source water for drinking water supplies. In contrast to the highly regulated domestic wastewater reuse practices in the U.S., not much is known regarding the human health risk associated with de facto reuse. Trinity River, Texas, a wastewater dominated river that serves as the main source of drinking water for Houston, is used as an exemplar to quantify the health risk associated with exposure to Cryptosporidium and norovirus through drinking water contaminated with wastewater effluents. The results show that the annual infection risks exceed the U.S. EPA recommended annual safe drinking water health risk of 10 −4 per person per year (pppy) for all scenarios considered. However, a comparison of the corresponding disease burdens with the World Health Organization's annual disease burden for safe drinking water of 10 −6 DALYs pppy indicates the disease burdens are mostly within the acceptable range of the guideline. This difference in risk interpretations reflects the difference in the fundamental definition of the two risk benchmarks as well as the gaps in data and risk models (including the conversion from infection to illness risk and expected DALY per infection). The results also imply that the current safe drinking water regulatory framework should be re-examined to best achieve the goal of human health protection. The outcomes of this study provide an overall picture of the potential risk associated with de facto reuse, which contributes to a better understanding of the practice and decisions in wastewater reuse in the U.S. and the rest of the world.

For many coastal regions around the world, recreational beach water quality is assessed using fecal indicator bacteria (FIB). However, the utility of FIB as indicators of recreational water illness (RWI) risk has been questioned, particularly in coastal settings with no obvious sources of human sewage. In this study we employed a source-apportionment quantitative microbial risk assessment (SA-QMRA) to assess RWI risk at a popular semi-enclosed recreational beach in Southern California (Baby Beach, City of Dana Point) with no obvious point sources of human sewage. Our SA-QMRA results suggest that, during dry weather, the median RWI risk at this beach is below the U.S. EPA recreational water quality criteria (RWQC) of 36 illness cases per 1000 bathers. During wet weather, the median RWI risk predicted by SA-QMRA depends on the assumed level of human waste associated with stormwater; the RWI risk is below the EPA RWQC illness risk benchmark 100% of the time provided that

Biofouling of seawater reverse osmosis (SWRO) membranes in desalination processes causes increased energy consumption and operating costs. In spite of pre-treatment systems, polymeric materials are deposited on the membranes along with bacteria and other particles. Bacteria grow and produce further polymers (extracellular polymeric substances; EPS) in situ forming a recalcitrant biofilm. Current membrane cleaning protocols aim to kill the bacteria but do not necessarily remove the associated polysaccharides which appear to reduce permeate permeability. Few studies have investigated the removal of both biofilm bacteria and associated polysaccharides. There is some evidence that compounds which produce free radicals can degrade polysaccharides; and the previous studies in our laboratory have suggested that they can reduce polysaccharide adhesion and the effects of membrane fouling. In this study, we compared the effect of two free radical producing systems with the currently accepted control agent, a biodegradable biocide, 2,2-dibromo-3-nitriloproprionamide (DBNPA). The free radical generating systems were sodium nitroprusside (SNP) that spontaneously releases nitric oxide free radical and a xanthine oxidase enzyme with a hypoxanthine substrate to release a superoxide radical. Experiments were conducted on the fouled membranes collected following membrane unit replacements at a full scale seawater desalination plant in Western Australia. Both free radical generating compounds improved permeate flow in a bench scale cross-flow RO system compared to the biocide without damaging membrane structures. The CLSM analysis suggested the biofilm was thinner but also less compact. A lectin bioassay supported the conclusion that the free radicals had a direct effect on the biofilm polysaccharides, not just the biofilm cells.

The effective tracking and purification of biological RNAs and RNA protein complexes is currently challenging. One promising strategy to simultaneously address both of these problems is to develop high-affinity RNA aptamers against taggable small molecule fluorophores. RNA Mango is a 39-nucleotide, parallel-stranded G-quadruplex RNA aptamer motif that binds with nanomolar affinity to a set of thiazole orange (TO1) derivatives while simultaneously inducing a 103-fold increase in fluorescence. We find that RNA Mango has a large increase in its thermal stability upon the addition of its TO1-Biotin ligand. Consistent with this thermal stabilization, RNA Mango can effectively discriminate TO1-Biotin from a broad range of small molecule fluorophores. In contrast, RNA Spinach, which is known to have a substantially more rigid G-quadruplex structure, was found to bind to this set of fluorophores, often with higher affinity than to its native ligand, 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI), and did not exhibit thermal stabilization in the presence of the TO1-Biotin fluorophore. Our data suggest that RNA Mango is likely to use a concerted ligand-binding mechanism that allows it to simultaneously bind and recognize its TO1-Biotin ligand, whereas RNA Spinach appears to lack such a mechanism. The high binding affinity and fluorescent efficiency of RNA Mango provides a compelling alternative to RNA Spinach as an RNA reporter system and paves the way for the future development of small fluorophore RNA reporter systems.

This study aims to evaluate the effects of a 12-week empowerment-based self-care programme on the cognitive, psychological and behavioural aspects of self-care, health-related quality of life and unplanned hospital service use of chronic heart failure patients and to estimate its cost-effectiveness in reducing unplanned service use.Randomized controlled trial with cost-effectiveness analysis.The study will recruit 236 community-dwelling Chinese patients with chronic heart failure from regional hospitals in Hong Kong. Patients will be randomized in blocks to attend either the 12-week empowerment-based self-care programme or a didactic education programme. Central to the empowerment-based model is that the patients are the primary decision makers in self-care and act autonomously to manage the disease. Collaborative approach and interactive teaching strategies are used to optimize patients' engagement and knowledge translation to real-life practice. Outcome evaluation on self-care maintenance and management, symptom perception, self-care confidence, self-care knowledge, health-related quality of life and health service use will take place at baseline, on completion of the programme and at 12 weeks thereafter. Generalized estimating equations and Cox regression examine the intervention effects. Bootstrapping technique will be conducted to examine the cost-effectiveness of the intervention. The study is formally funded in December 2016.Chronic heart failure is associated with high level of hospital admissions, of which 40% is avoidable through effective self-care. Although self-care confidence is known to improve self-care, no structured educative method has been developed to enhance this attribute. This study addresses this research gap with a stringent application of theory and research design.The study illustrates the application of the empowerment model for self-care enhancement in patients with chronic heart failure. The findings will inform the ways and values of this care model to enhance the disease management of this expanding clinical cohort.目的: 本研究旨在评估一个12周的基于赋权的自我护理项目对于认知、心理和行为方面的自我护理,健康相关生活质量和慢性心力衰竭病人的计划外医院运送服务和估算其成本效益减少计划外的运送服务使用。 研究设计: 成本效益分析的随机对照实验。 方法: 本研究将从香港地区医院招募236名居住在社区的中国慢性心力衰竭患者。患者将被随机分组,参加为期12周的基于授权自我护理项目或说教教育项目。以授权为基础的模式的核心是,患者是自我护理的主要决策者,并自主采取行动来管理疾病。采用协作方法和交互式教学策略以优化患者的参与和将知识转化为现实生活中的实践。将在基线、项目完成时和在12周后对自我护理维护和管理、症状感知、自我护理信心、自我护理知识、健康相关生活质量和健康服务使用的结果进行评估。广义评估方程和Cox回归分析用于检验干预效果。引导技术将用于检查干预的成本效益。该研究于2016年12月正式获得资金赞助。 讨论: 慢性心力衰竭直接相关于高住院率,其中40%的住院率可通过有效的自我护理避免。尽管自我护理信心可以改善自我照顾,但是还没有一种结构化的教育方法可以增强这种自信。本研究通过严格的理论应用和研究设计来弥补这一研究空白。 影响: 本研究阐释了授权模式在增强慢性心力衰竭患者自我照顾中的应用。本研究结果将为该护理模式的方法和价值提供信息,以提高逐渐扩大的临床治疗患者队伍的疾病管理。.

Background Nigeria faces health workforce challenges and poor population health indices resulting from disparities in health worker densities by geographical locations and levels of health care delivery. Nigeria is constantly reforming its health system with the primary aim of having the right number of health workers in the right place at the right time to meet the population’s health needs. The majority of primary health facilities in the country are staffed using perceived needs. The Workload Indicators of Staffing Need (WISN) tool developed by the World Health Organization is used to determine staffing requirements for facilities. Methods The WISN tool was used in assessing the staffing requirements for nurses/midwives and community health practitioners in 26 primary health facilities in Port Harcourt City Local Government Area (PHALGA) and Obio Akpor Local Government Area (OBALGA). Documents were reviewed to obtain information on working conditions and staffing, and interviews conducted with key informants in 12 randomly selected facilities. We supported an expert working group that comprised of nurses/midwives and community health practitioners to identify workload components and activity standards and validate both. We also retrieved workload data from January 1–December 31, 2015 from the national district health information system. Results Findings showed varying degrees of shortages and inequitable distribution of health workers. Health facilities in PHALGA had a WISN ratio of 0.63 and a shortage of 31 nurses/midwives. There was also a shortage of 12 community health practitioners with a WISN ratio of 0.85. OBALGA had a shortage of 50 nurses/midwives and 24 community health practitioners; and WISN ratios of 0.60 and 0.79 for nurses/midwives and community health practitioners respectively. Conclusion Our findings provide evidence for policies that will help Nigeria improve the population’s access to quality health services and reduce inequities in distribution of the health workforce. Evidence-based health workforce planning and redistribution using WISN should be institutionalized. Review of scopes of practice of health workforce should be conducted periodically to ensure that the scope of practice matches the training received by the specific cadres and those skills are used to deliver quality services.

International audience; Several turn-on RNA aptamers that activate small molecule fluorophores have been selected in vitro. Among these, the ~30 nucleotide Mango-III is notable because it binds the thiazole orange derivative TO1-Biotin with high affinity and fluoresces brightly (quantum yield 0.55). Uniquely among related aptamers, Mango-III exhibits biphasic thermal melting, characteristic of molecules with tertiary structure. We report crystal structures of TO1-Biotin complexes of Mango-III, a structure-guided mutant Mango-III(A10U), and a functionally reselected mutant iMango-III. The structures reveal a globular architecture arising from an unprecedented pseudoknot-like connectivity between a G-quadruplex and an embedded non-canonical duplex. The fluorophore is restrained into a planar conformation by the G-quadruplex, a lone, long-range trans-Watson-Crick pair (whose A10U mutation increases quantum yield to 0.66), and a pyrimidine perpendicular to the nucleobase planes of those motifs. The improved iMango-III and Mango-III(A10U) fluoresce ~50% brighter than enhanced green fluorescent protein, making them suitable tags for live cell RNA visualization.

PURPOSE: To improve care coordination for complex cancers, it is critical to establish a more nuanced understanding of the types of providers involved. As the number of provider types increases, strategies to support cancer care coordination must adapt to a greater variety of information needs, communication styles, and treatment strategies. METHODS: We categorized providers into 11 types, using National Provider Identifier specialties. Using Medicare claims, we counted the number of unique combinations of provider types billed during preoperative, operative, and postdischarge care for colon cancer surgery and assessed how this count varies across hospitals. The study included 70,567 beneficiaries in fee-for-service Medicare A and B for 6 months before and 60 days after an admission for colectomy for colon cancer between 2008 and 2011. RESULTS: We observed 1,554 preoperative provider-type combinations, 975 operative combinations, and 1,571 postdischarge combinations. The three most common combinations in the preoperative phase were general medicine only, other medical specialists only, and general medicine and other medical specialists. In the operative phase, the three most common combinations were primary surgery, anesthesiology, and pathology; general medicine, other medical specialists, radiology, primary surgery, anesthesiology, and pathology; and other medical specialists, radiology, primary surgery, anesthesiology, and pathology. In the postdischarge phase, the three most common combinations were general medicine, general medicine and other medical specialists, and general medicine and oncology. On average, each hospital had 15 preoperative, 11 operative, and 15 postoperative combinations. High-volume, larger, teaching, urban, and noncritical access hospitals had more combinations in all phases. CONCLUSION: Many provider-type combinations are involved in colon cancer surgery care. Substantial variation exists across hospitals types, suggesting that certain hospitals need additional resources and more flexible infrastructure to coordinate care.

Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells., Many aptamer-fluorophore complexes suffer from low quantum yield and low extinction coefficients limiting their usability. Here the authors isolate new Mango aptamers with improved fluorescent properties for both fixed and live-cell imaging by using competitive ligand binding microfluidic selection.

We describe a simple bioinformatics method for biomarker discovery that is based on the analysis of global transcript levels in a population of inbred mouse strains showing variation for disease-related traits. This method has advantages such as controlled environment and accessibility to heart and plasma tissue in the preclinical selection stage. We illustrate the approach by identifying candidate heart failure (HF) biomarkers by overlaying mouse transcriptome and clinical traits from 91 Hybrid Mouse Diversity Panel (HMDP) inbred strains and human HF transcriptome from the Myocardial Applied Genomics Network (MAGNet) consortium. We found that some of the top differentially expressed genes correlated with known human HF biomarkers, such as galectin-3 and tissue inhibitor of metalloproteinase 1. Using ELISA assays, we investigated one novel candidate, Glycoprotein NMB, in a mouse model of chronic β-adrenergic stimulation by isoproterenol (ISO) induced HF. We observed significantly lower GPNMB plasma levels in the ISO model compared to the control group (p-value = 0.007). In addition, we assessed GPNMB plasma levels among 389 HF cases and controls from the METabolic Syndrome In Men (METSIM) study. Lower levels of GPNMB were also observed in patients with HF from the METSIM study compared to non-HF controls (p-value < 0.0001). In summary, we have identified several candidate biomarkers for HF using the cardiac transcriptome data in a population of mice that may be directly relevant and applicable to human populations.

Microcystin (MC), a hepatotoxin that is associated with cyanobacterial blooms in freshwater lakes, threatens the quality of drinking water resources. Biodegradation of MC using biofiltration is emerging as a cost-effective solution for drinking water treatment. This study reports isolation of five MC-degrading microbial consortia and investigation of their community structure and kinetics in the presence or absence of a readily-bioavailable organic carbon source. The results indicated that the presence of a bioavailable organic carbon source caused: (1) the proliferation of community members previously unobserved in each consortium cultured without ethanol, (2) a shift in abundance of representative taxa, (3) a fluctuation in genera affiliated with MC-biodegradation, and, (4) a unique response in simulated diversity among consortia. These changes to each microbial consortium were paralleled by a significant decline in MC degradation kinetics. Overall, this study highlights the importance of integrating environmental conditions into the design and operation of biofiltration systems for MC biodegradation.

The discovery of cheap and eco-friendly functional materials for the removal of anionic heavy metal ions is still challenging in the treatment of heavy metal-contaminated water. Herein, a new poly(allyltrimethylammonium) grafted chitosan and biochar composite (PATMAC-CTS-BC) was introduced for the removal of selenate (SeO42−) in water. Results suggest that the PATMAC-CTS-BC showed a rapid removal of SeO42− with efficiency of >97% within 10 min and it followed a pseudo-second-order model. High capacity of SeO42− adsorption by the composite was achieved, with maximum value of 98.99 mg g−1 based on Langmuir model, considerably higher than most of reported adsorbents. The thermodynamic results reflected the spontaneous and exothermic nature of SeO42− adsorption onto the composite. The composite could be applied at a wide initial pH range (2–10) with high removal efficiency of SeO42− because of permanent positive charges of quaternary ammonium groups (=N+–). The removal mechanisms of SeO42− were mainly attributed to electrostatic interactions with =N+– and protonated –NH3+ groups, and redox-complexation interactions with –NH2, –NH–, and –OH groups. Besides SeO42−, the hexavalent chromium (Cr2O72−) was considered as example to further demonstrate the anion removal capability of cationic hydrogel-BC composite. The study outcomes open up new opportunities to efficiently remove anionic heavy metal ions (e.g., SeO42− and Cr2O72−) in water using these materials.

Bacterial biofilm formation, the main cause of membrane biofouling, is a crucial issue for membrane separation. Biofilm production is regulated by quorum sensing (QS) systems where bacteria secrete auto-inducers to communicate with neighboring bacteria. This research identified that several marine bacteria isolated from a desalination plant produced a low molecular weight auto-inducer 1 (AI-1) signaling molecule. AI-1 production in the mixed culture of the four different biofilm-forming marine bacteria was greater than in individual bacterial cultures. The QS inhibiting compounds, vanillin and cinnamaldehyde at 1200 mg/L significantly reduced biofilm formed by these marine bacteria by more than 79% and 70%, respectively in a microtiter plate assay. Anti-biofilm capabilities of vanillin and cinnamaldehyde were further assessed in a reverse osmosis membrane bio-monitoring system using mixed bacterial cultures and native uncultured bacterial communities in natural seawater. Confocal microscopy showed vanillin (1200 mg/L) significantly reduced biofilm extracellular polysaccharides and dead cells on the membrane surface (> 40%, > 20%). These results indicate that QS inhibitors have the potential to remediate membrane biofouling.

Marine beaches are important recreational and economic resources in Brazil, but the beaches' water quality is negatively impacted by the discharge of domestic sewage effluent. The occurrence of diarrheagenic Escherichia coli among the E. coli isolated from three Brazilian marine beaches was investigated. Multiplex and single step PCR were used to screen 99 E. coli isolates for ten target toxin genes. Six toxin genes, stx1, eae, estp, esth, astA, and bfpA, were identified in 1% to 35% of the isolates. A quantitative microbial risk assessment (QMRA) of human exposure to diarrheagenic E. coli during marine recreation was carried out. The results indicated that the diarrheagenic E. coli risk is well below the U.S. EPA's recommended daily recreational risk benchmark. However, the overall recreational health risk due to all pathogens in the water could be much higher and exceeded the U.S. EPA's benchmark.

The diffusion of health information exchange (HIE), in which clinical data are electronically linked to patients in many different care settings, is a top priority for policy makers. To drive HIE, community and state efforts were federally funded to broadly engage providers in exchanging data in ways that improved patient care. To assess the current landscape, we conducted a national survey of community and state HIE efforts soon after federal funding ended. We found 106 operational HIE efforts that, as a group, engaged more than one-third of all US providers in 2014. However, the number of operational HIE efforts is down from 119 in 2012, representing the first decline observed since the tracking of these efforts began in 2006. Only half of operational efforts reported being financially viable, and all efforts reported a variety of barriers to continuation. These findings raise important questions about whether the current vision for HIE efforts will allow for the broad exchange of clinical data, or whether alternative approaches would be more successful.

Polymerase chain reaction (PCR)-based methods have been developed and increasingly used for rapid and sensitive detection of pathogens in water samples to better protect public health. A propidium monoazide (PMA) pretreatment can help to differentiate between viable and dead cells, but the photoactivation of PMA normally requires the use of an energy-consuming halogen light, which is not suitable for off-the-grid applications. Herein, we investigate sunlight as an alternative light source. Our results suggest that sunlight can successfully activate PMA, and the sunlight-activated PMA pretreatment can effectively reduce the amplification of DNA derived from dead cells in PCR assays. Potentially, a sunlight-activated PMA pretreatment unit can be integrated into a lab-on-a-chip PCR device for off-the-grid microbial detection and quantification.

Background: Because specialty care accounts for half of Medicare expenditures, improving its value is critical to the success of Medicare accountable care organizations (ACOs) in curbing spending growth. However, whether ACOs have reduced low-value specialty care without compromising use of high-value services remains unknown. Methods and Results: Using national Medicare data, we identified 2 cohorts: beneficiaries for whom the value of coronary revascularization is lower (those with ischemic heart disease without angina, congestive heart failure, or recent admission for acute myocardial infarction) and beneficiaries for whom its value is higher (those with recent acute myocardial infarction admission). We then determined the provider groups who cared for the cohorts, distinguishing between those participating (n=298) and those not participating in a Medicare ACO (1329). After measuring the provider groups’ use of coronary artery bypass grafting and percutaneous coronary intervention among the 2 cohorts, we fit multivariable models to test the statistical significance of rates of change in low- and high-value revascularization after ACO participation. During the pre-ACO period, participating and nonparticipating provider groups had similar rates of low- and high-value revascularization. Our multivariable model results show that rates of change for low- and high-value coronary revascularization were not altered by a provider group’s participation in a Medicare ACO (lower value: difference, −0.04 per year; 95% confidence interval, −0.11 to 0.03; higher value: difference, 0.96 per year; 95% confidence interval, −0.46 to 2.4). Conclusions: We found no association between provider group participation in a Medicare ACO and use of low- or high-value coronary revascularization.

Regenerative medicine holds great promise for both degenerative diseases and traumatic tissue injury which represent significant challenges to the health care system. Hearing loss, which affects hundreds of millions of people worldwide, is caused primarily by a permanent loss of the mechanosensory receptors of the inner ear known as hair cells. This failure to regenerate hair cells after loss is limited to mammals, while all other non-mammalian vertebrates tested were able to completely regenerate these mechanosensory receptors after injury. To understand the mechanism of hair cell regeneration and its association with regeneration of other tissues, we performed a guided mutagenesis screen using zebrafish lateral line hair cells as a screening platform to identify genes that are essential for hair cell regeneration, and further investigated how genes essential for hair cell regeneration were involved in the regeneration of other tissues. We created genetic mutations either by retroviral insertion or CRISPR/Cas9 approaches, and developed a high-throughput screening pipeline for analyzing hair cell development and regeneration. We screened 254 gene mutations and identified 7 genes specifically affecting hair cell regeneration. These hair cell regeneration genes fell into distinct and somewhat surprising functional categories. By examining the regeneration of caudal fin and liver, we found these hair cell regeneration genes often also affected other types of tissue regeneration. Therefore, our results demonstrate guided screening is an effective approach to discover regeneration candidates, and hair cell regeneration is associated with other tissue regeneration., Identifying regenerative genes in non-mammalian vertebrates A study on zebrafish has genetically screened 254 genes and identified 7 genes implicated in the development and regeneration of hair cells and other tissues. Humans and other mammals cannot regrow hair cells—inner-ear sensory receptors that enable hearing—whereas non-mammalian vertebrates, including zebrafish, can regrow these following injury. Researchers from the United States, led by the National Institutes of Health’s Shawn Burgess, screened adult zebrafish for genes active during the regeneration of inner-ear epithelium. The researchers then produced zebrafish without these genes to study their functions. The studies tested 254 genes known to respond during regeneration, and identified seven specifically impacting regeneration. Most of these seven genes also functioned in liver and fin tissue regeneration. Understanding the mechanisms of these genes may enable future research into regenerative therapies in humans.

Isoproterenol is used widely for inducing heart failure in mice. Isoproterenol is a nonselective beta-adrenergic agonist. The acute model mimics stress-induced cardiomyopathy. The chronic model mimics advanced heart failure in humans. In this chapter, we describe a protocol that we used to induce heart failure in 100+ strains of inbred mice. Techniques on surgical pump implantation and echocardiography are described in detail. We also discuss the impact of drug dosage, duration, mortality, age, gender, and strain on cardiac remodeling responses. The success of model creation may be assessed by echocardiogram or molecular markers. This chapter may be relevant to those who are interested in using this heart failure model.

As more countries engage in water reuse, either intended or de facto, there is an urgent need to more comprehensively evaluate resulting environmental and public health concerns. While antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are increasingly coming under the spotlight, as emerging contaminants, existing water reuse regulations and guidelines do not adequately address these concerns. This perspectives paper seeks to frame the various challenges that need to be resolved to identify meaningful and realistic target types and levels of antibiotic resistance benchmarks for water reuse. First, there is the need for standardized and agreed-upon methodologies to identify and quantify ARB and ARGs. Second, even if methodologies are available, identifying which ARB and ARGs to monitor that would best relate to the occurrence of disease burden remains unknown. Third, a framework tailored to assessing the risks associated with ARB and ARGs during reuse is urgently needed. Fourth, similar to protecting drinking water sources, strategies to prevent dissemination of ARB and ARGs via wastewater treatment and reuse are required to ensure that appropriate barriers are emplaced. Finally, current wastewater treatment technologies could benefit from modification or retrofit to more effectively remove ARB and ARGs while also producing a high quality product for water and resource recovery. This perspectives paper highlights the need to consider ARB and ARGs when evaluating the overall safety aspects of water reuse and ways by which this may be accomplished. Published version

Objective To identify factors that impede payer engagement in a health information exchange (HIE), along with organizational and policy strategies that might effectively address the impediments.Materials and Methods Qualitative analysis of semi-structured interviews with leaders from 17 varied payer organizations from across the country (e.g., large, national payers; state Blues plans; local Medicaid managed care plans).Results We found a large gap between payers’ vision of what optimal HIE should be and the current approach to HIE in the United States. Notably, payers sought to be active participants in HIE efforts – both providing claims data and accessing clinical data to support payer HIE use cases. Instead, payers were often asked by HIE efforts only to provide financial support without the option to participate in data exchange, or, when given the option, their data needs were secondary to those of providers.Discussion Efforts to engage payers in pursuit of more robust and sustainable HIE need to better align their value proposition with payer HIE use cases. This will require addressing provider concerns about payer access to clinical data. Policymakers should focus on creating the conditions for broader payer engagement by removing common obstacles, such as low provider engagement in HIE.Conclusion Despite variation in the extent to which payers engaged with current HIE efforts, there was agreement on the vision of optimal HIE and the facilitators of greater payer engagement. Specific actions by those leading HIE efforts, complemented by policy efforts nationally, could greatly increase payer engagement and enhance HIE sustainability.

RNA plays important roles in cellular processes, but RNA-protein complexes are notoriously hard to isolate and study. We compare and contrast existing RNA- and protein-purification strategies with the potential of new RNA-tagging systems such as RNA Spinach and RNA Mango. Each RNA aptamer binds a small fluorophore, resulting in a highly fluorescent complex that is thousands of times brighter than the unbound fluorophore. Provided that the aptamer binding affinity is high enough, derivatized dyes can be used in conjunction with these aptamers to purify RNA complexes while simultaneously using their intrinsic fluorescence to track the complex of interest. The known strengths and weakness of these RNA tagging systems are discussed.

Expansion of mesenchymal stem cells (MSCs) and maintenance of their self-renewal capacity in vitro requires specialized robust cell culture systems. Conventional approaches using animal-derived or artificial matrices and a cocktail of growth factors have limitations such as consistency, scalability, pathogenicity, and loss of MSC phenotype. Herein, we report the use of all-carbon 3-D single- and multiwalled carbon nanotube scaffolds (SWCNTs and MWCNTs) as artificial matrices for long-term maintenance and expansion of human MSCs. Three-dimensional SWCNT and MWCNT scaffolds were fabricated using a novel radical initiated thermal cross-linking method that covalently cross-links CNTs to form 3-D macroporous all-carbon architectures. Adipose-derived human MSCs showed good cell viability, attachment, proliferation, and infiltration in MWCNT and SWCNT scaffolds comparable to poly(lactic-co-glycolic) acid (PLGA) scaffolds (baseline control). ADSCs retained stem cell phenotype after 30 days and satisfied the International Society for Cellular Therapy's (ISCT) minimal criteria for MSCs. Post expansion, (1) ADSCs showed in vitro adherence to tissue culture polystyrene (TCPS); (2) MSC surface antigen expression [CD14(-), CD19(-), CD34(-), CD45(-), CD73(+), CD90(+), CD105(+)]; and (3) trilineage differentiation into osteoblasts, adipocytes, and chondrocytes. Results show that cross-linked 3-D MWCNTs and SWCNTs scaffolds are suitable for ex vivo expansion and maintenance of MSCs for therapeutic applications. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1927-1939, 2017.

Pseudo-nitzschia blooms often occur in coastal and open ocean environments, sometimes leading to the production of the neurotoxin domoic acid that can cause severe negative impacts to higher trophic levels. Increasing evidence suggests a close relationship between phytoplankton bloom and bacterial assemblages, however, the microbial composition and succession during a bloom process is unknown. Here, we investigate the bacterial assemblages before, during and after toxic and non-toxic Pseudo-nitzschia blooms to determine the patterns of bacterial succession in a natural bloom setting. Opportunistic sampling of bacterial community profiles were determined weekly at Santa Cruz Municipal Wharf by 454 pyrosequencing and analyzed together with domoic acid levels, phytoplankton community and biomass, nutrients and temperature. We asked if the bacterial communities are similar between bloom and non-bloom events and if domoic acid or the presence of toxic algal species acts as a driving force that can significantly structure phytoplankton-associated bacterial communities. We found that bacterial diversity generally increases when Pseudo-nitzschia numbers decline. Furthermore, bacterial diversity is higher when the low-DA producing P. fraudulenta dominates the algal bloom while bacterial diversity is lower when high-DA producing P. australis dominates the algal bloom, suggesting that the presence of algal toxin can structure bacterial community. We also found bloom-related succession patterns among associated bacterial groups; Gamma-proteobacteria, were dominant during low toxic P. fraudulenta blooms comprising mostly of Vibrio spp., which increased in relative abundance (6-65%) as the bloom progresses. On the other hand, Firmicutes bacteria comprising mostly of Planococcus spp. (12-86%) dominate during high toxic P. australis blooms, with the bacterial assemblage showing the same bloom-related successional patterns in three independent bloom events. Other environmental variables such as nitrate and phosphate and temperature appear to influence some low abundant bacterial groups as well. Our results suggest that phytoplankton-associated bacterial communities are strongly affected not just by phytoplankton bloom in general, but also by the type of algal species that dominates in the natural bloom.

Phosphoribosyl pyrophosphate synthetase-1 (PRPS1) is a key enzyme in nucleotide biosynthesis and mutations in PRPS1 are found in several human diseases including nonsyndromic sensorineural deafness, Charcot-Marie-Tooth disease-5 and Arts Syndrome. We utilized zebrafish as a model to confirm that mutations in PRPS1 result in phenotypic deficiencies in zebrafish similar to those in the associated human diseases. We found two paralogs in zebrafish, prps1a and prps1b and characterized each paralogous mutant individually as well as the double mutant fish. Zebrafish prps1a mutants and prps1a;prps1b double mutants showed similar morphological phenotypes with increasingly severe phenotypes as the number of mutant alleles increased. Phenotypes included smaller eyes and reduced hair cell numbers, consistent with the optic atrophy and hearing impairment observed in human patients. The double mutant also showed abnormal development of primary motor neurons, hair cell innervation and reduced leukocytes, consistent with the neuropathy and recurrent infection of the human patients possessing the most severe reductions of PRPS1 activity. Further analyses indicated the phenotypes were associated with a prolonged cell cycle likely resulting from reduced nucleotide synthesis and energy production in the mutant embryos. We further demonstrated the phenotypes were caused by delays in the tissues most highly expressing the prps1 genes.

Objective: To compare the psychological health of men with partners who have postpartum depression (PPD; index group) with that of men with partners without PPD (comparison group). Method: Using a cross-sectional survey, psychological symptoms and disturbances of index group men (n = 58) and comparison group men (n = 116) were compared. Validated self-report measures were used to assess five key areas of mental health: depression, anxiety, non-specific psychological impairment, aggression and alcohol use. Results: Index group men had more symptoms of depression, aggression and nonspecific psychological impairment, and had higher rates of depressive disorder, nonspecific psychological problems and problem fatigue than comparison group men. Index group men were also more likely to have three or more comorbid psychological disturbances. There was no difference between the groups on measures of anxiety and alcohol use. Conclusions: Although many men in the postnatal period experience a variety of mental health problems, those who have a partner with PPD are themselves at increased risk for experiencing psychological symptoms and disturbances. Differentiation of psychological syndromes is important; higher rates of depressive disorder, non-specific psychological problems and problem fatigue were found, but rates of anxiety disorder and hazardous alcohol use did not differ between the groups. More attention from health professionals to men's mental health in the postnatal period may be beneficial to the entire family system.

The purpose of this investigation was to identify a quantitative dose-response relationship for enhancing maximal oxygen consumption (VO2max) in healthy sedentary older adults after controlled endurance training.This meta-analysis of controlled clinical trials included 1257 exercisers and 845 controls with a mean age of 67.45 ± 5.25 years. Effect sizes were calculated for training-induced VO2max changes. Different training regimens were analyzed and compared. The weighted net change of the mean VO2max values showed a significant increase of 3.78 ml/kg per min (95% confidence interval = 3.29 to 4.27; p 0.0001) in response to aerobic training. Interstudy differences in VO2max changes were significantly related to exercise intensity, and explained approximately 11% of the variance of the VO2max responses. VO2max improved significantly at 35%-50% heart rate reserve (HRR) and continued improving at a greater rate with increasing "dose". The largest VO2max-improvement adaptation was achieved with a mean intensity of 66%-73% HRR. The magnitudes of the VO2max adaptation are identical to exercise at 57%-65% HRR and at 75%-80% HRR. Higher intensity doses more than 75-80% HRR did not lead to greater enhancement of VO2max improvements but, conversely, resulted in large declines.Our data provide quantitative insight into the magnitude of VO2max alterations as affected by exercise intensity, duration, frequency, and program length. The shapes of the dose-response curves are not simply linear, but with many similar trends and noteworthy characteristics. Aerobic training at a mean intensity of 66%-73% HRR with 40-50 min per session for 3-4 day/week for 30-40 weeks appears to be effective and optimal for maximum cardiorespiratory benefits in healthy sedentary older adults.

Because RNA lacks strong intrinsic fluorescence, it has proven challenging to track RNA molecules in real time. To address this problem and to allow the purification of fluorescently tagged RNA complexes, we have selected a high affinity RNA aptamer called RNA Mango. This aptamer binds a series of thiazole orange (fluorophore) derivatives with nanomolar affinity, while increasing fluorophore fluorescence by up to 1,100-fold. Visualization of RNA Mango by single-molecule fluorescence microscopy, together with injection and imaging of RNA Mango/fluorophore complex in C. elegans gonads demonstrates the potential for live-cell RNA imaging with this system. By inserting RNA Mango into a stem loop of the bacterial 6S RNA and biotinylating the fluorophore, we demonstrate that the aptamer can be used to simultaneously fluorescently label and purify biologically important RNAs. The high affinity and fluorescent properties of RNA Mango are therefore expected to simplify the study of RNA complexes.

Background: We are using genetics to identify genes specifically involved in hearing regeneration. In a large-scale genetic screening, we identified mgat5a, a gene in the N-glycosylation biosynthesis pathway whose activity negatively impacts hair cell regeneration. Methods: We used a combination of mutant analysis in zebrafish and a hair cell regeneration assay to phenotype the loss of Mgat5a activity in zebrafish. We used pharmacological inhibition of N-glycosylation by swansonine. We also used over-expression analysis by mRNA injections to demonstrate how changes in N-glycosylation can alter cell signaling. Results: We found that mgat5a was expressed in multiple tissues during zebrafish embryo development, particularly enriched in neural tissues including the brain, retina, and lateral line neuromasts. An mgat5a insertional mutation and a CRISPR/Cas9-generated truncation mutation both caused an enhancement of hair cell regeneration which could be phenocopied by pharmacological inhibition with swansonine. In addition to hair cell regeneration, inhibition of the N-glycosylation pathway also enhanced the regeneration of lateral line axon and caudal fins. Further analysis showed that N-glycosylation altered the responsiveness of TGF-beta signaling. Conclusions: The findings from this study provide experimental evidence for the involvement of N-glycosylation in tissue regeneration and cell signaling.