Public Interest in a Potentially Harmful, Non-Evidence-Based "Wellness" Practice: Cross-Sectional Analysis of Perineum Sunning.
Perineum sunning/tanning is a potentially harmful yet popular new health trend cultivated by a viral social media post, famous public figures, and subsequent media coverage. Our primary objective is to evaluate public interest in perineum sunning/tanning. Using an observational study design, we extracted data from Google Trends for the terms "perineum sunning," "perineum tanning," "Metaphysical Meagan," and "Josh Brolin"; and Twitter (via SproutSocial) for "perineum sunning" and "perineum tanning" from November 1, 2019, to December 31, 2019. UberSuggest was used to investigate monthly search volumes and user engagement. We used data from Google Trends and Twitter to construct autoregressive integrated moving average (ARIMA) models to forecast public interest in perineum sunning and perineum tanning had the post on social media never occurred. Next, we performed an integral function to calculate the cumulative increase in "perineum tanning" from the day after the post occurred to the end of the year as the area between the forecasted values and the actual values. Using Welch t tests, we compared forecasted and actual values for "perineum sunning" and "perineum tanning" using Twitter and Google Trends data over 1-, 2-, and 4-week periods after the social media post to determine if the increased volumes were statistically significant over time. Lastly, we monitored Google Trends for "perineum sunning" and "perineum tanning" through September 30, 2020, to capture trends during the summer months. Before the Instagram post went viral, there was no search interest in perineum sunning. ARIMA modeling for perineum tanning forecasted no increase in searches (0.00) if the post had not gone viral, while actual interest conveyed a relative cumulative increase of 919.00% from the day the post went viral through December 31, 2020. The term "perineum sunning" was mentioned on average 804 (SD 766.1) times daily for this 7-day period, which was also significantly higher than predicted (P≤.03), totaling 5628 tweets for these 7 days. The increased volume of tweets and relative search interest from Google Trends remained significantly higher for both terms over the 1-, 2-, and 4-week intervals. User engagement showed that nearly 50% of people who searched for "perineum sunning" were likely to click a returned link for more information. Continued observance of search interest in perineum sunning demonstrated interest spikes in the summer months, June and July 2020. Google Trends and Twitter data demonstrated that one social media post claiming non-evidence-based health benefits of regular sun exposure-without the use of sunscreen-generated significant public interest. Medical journals, dermatologists, and other health care professionals are obligated to educate and correct public misperceptions about viral wellness trends such as perineum sunning.


Sharpening and formation: two distinct neuronal mechanisms of repetition priming.
The repetition of an object stimulus results in faster and better recognition of this object (repetition priming). This phenomenon is neuronally associated with a reduced firing rate of neurons (repetition suppression). It has been interpreted as a sharpening mechanism within the cell assembly representing the object. In the case of an unfamiliar stimulus for which no object representation exists, the repetition of the stimulus results in an increase in the firing rate (repetition enhancement).It has been hypothesized that this increase reflects the formation of a cortical object representation. We aimed to investigate cortical object representations as well as repetition suppression and enhancement by means of the steady-state visual evoked potential (SSVEP) in the healthy human brain. To that end, we used a repetition paradigm with familiar and unfamiliar objects, each presented with 12-Hz flicker, producing an oscillatory brain response at the same frequency (i.e. an SSVEP). Results showed significantly smaller SSVEP amplitudes for repeated familiar objects compared to their first presentation (repetition suppression). For unfamiliar objects, SSVEP amplitudes increased with stimulus repetition (repetition enhancement). Source reconstruction revealed inferior temporal regions as generators for the repetition suppression effect, probably reflecting a sharpening mechanism within the cortical representations of the constituting features of an object. In contrast, repetition enhancement was localised in the superior parietal lobe, possibly reflecting the formation of a structural object representation. Thus, the mechanisms underlying repetition priming (i.e. sharpening and formation) depend on the semantic content of the incoming information.


Inverse Design Strategies for 3D Surfaces Formed by Mechanically Guided Assembly.
Deterministic transformations of 2D patterns of materials into well-controlled 3D mesostructures serve as the basis for manufacturing methods that can bypass limitations of conventional 3D micro/nanofabrication. Here, guided mechanical buckling processes provide access to a rich range of complex 3D mesostructures in high-performance materials, from inorganic and organic semiconductors, metals and dielectrics, to ceramics and even 2D materials (e.g., graphene, MoS2 ). Previous studies demonstrate that iterative computational procedures can define design parameters for certain targeted 3D configurations, but without the ability to address complex shapes. A technical need is in efficient, generalized inverse design algorithms that directly yield sets of optimized parameters. Here, such schemes are introduced, where the distributions of thicknesses across arrays of separated or interconnected ribbons provide scalable routes to 3D surfaces with a broad range of targeted shapes. Specifically, discretizing desired shapes into 2D ribbon components allows for analytic solutions to the inverse design of centrally symmetric and even general surfaces, in an approximate manner. Combined theoretical, numerical, and experimental studies of ≈20 different 3D structures with characteristic sizes (e.g., ribbon width) ranging from ≈200 µm to ≈2 cm and with geometries that resemble hemispheres, fire balloons, flowers, concave lenses, saddle surfaces, waterdrops, and rodents, illustrate the essential ideas.


Health-related quality of life and functional status quality indicators for older persons with multiple chronic conditions.
To explore central challenges with translating self-reported measurement tools for functional status and health-related quality of life (HRQOL) into ambulatory quality indicators for older people with multiple chronic conditions (MCCs). Review. Sources including the National Quality Measures Clearinghouse and National Quality Forum were reviewed for existing ambulatory quality indicators relevant to functional status, HRQOL, and people with MCCs. Seven informants with expertise in indicators using functional status and HRQOL. Informant interviews were conducted to explore knowledge about these types of indicators, particularly usability and feasibility. Nine important existing indicators were identified in the review. For process, identified indicators addressed whether providers assessed functional status; outcome indicators addressed quality of life. In interviews, informants agreed that indicators using self-reported data were important in this population. Challenges identified included concerns about usability due to inability to discriminate quality of care adequately between organizations and feasibility concerns regarding high data collection burden, with a correspondingly low response rate. Validity was also a concern because evidence is mixed that healthcare interventions can improve HRQOL or functional status for this population. As a possible first step, a structural standard could be systematic collection of these measures in a specific setting. Although functional status and HRQOL are important outcomes for older people with MCCs, few relevant ambulatory quality indicators exist, and there are concerns with usability, feasibility, and validity. Further research is needed on how best to incorporate these outcomes into quality indicators for people with MCCs.


Modeling the site-specific variation of selection patterns along lineages.
The unambiguous footprint of positive Darwinian selection in protein-coding DNA sequences is revealed by an excess of nonsynonymous substitutions over synonymous substitutions compared with the neutral expectation. Methods for analyzing the patterns of nonsynonymous and synonymous substitutions usually rely on stochastic models in which the selection regime may vary across the sequence but remains constant across lineages for any amino acid position. Despite some work that has relaxed the constraint that selection patterns remain constant over time, no model provides a strong statistical framework to deal with switches between selection processes at individual sites during the course of evolution. This paper describes an approach that allows the site-specific selection process to vary along lineages of a phylogenetic tree. The parameters of the switching model of codon substitution are estimated by using maximum likelihood. The analysis of eight HIV-1 env homologous sequence data sets shows that this model provides a significantly better fit to the data than one that does not take into account switches between selection patterns in the phylogeny at individual sites. We also provide strong evidence that the strength and the frequency of occurrence of selection might not be estimated accurately when the site-specific variation of selection regimes is ignored.


Venous thromboembolism and risk of idiopathic interstitial pneumonia: a nationwide study.
Idiopathic interstitial pneumonia is characterized by pulmonary fibrosis and high mortality. We examined the association between ever-diagnosed venous thromboembolism and risk of incident idiopathic interstitial pneumonia. Venous thromboembolism was taken as a proxy for a procoagulant state in an individual. We conducted a study of the entire Danish population from 1980 through 2007, comprising 7.4 million individuals. Incident idiopathic interstitial pneumonia, ever-diagnosed venous thromboembolism, and use of prescription anticoagulants were drawn from national Danish registries. Age-standardized incidence rates per 10,000 person-years for idiopathic interstitial pneumonia were higher among those ever diagnosed with venous thromboembolism (1.8; n = 158,676), pulmonary embolism (2.8; n = 70,586), and deep venous thrombosis only (1.2; n = 88,090), than among control subjects (0.8; n = 7,260,278). Multivariate-adjusted hazard ratios for idiopathic interstitial pneumonia were 1.8 (95% confidence interval [CI], 1.7-1.9) in those ever diagnosed with venous thromboembolism, 2.4 (95% CI, 2.3-2.6) in those ever diagnosed with pulmonary embolism, and 1.3 (95% CI, 1.2-1.4) in those ever diagnosed with deep venous thrombosis only, compared with control subjects. Corresponding hazard ratios in those ever diagnosed with venous thromboembolism stratified in those ever and never treated with anticoagulants were 1.4 (95% CI, 1.2-1.6) and 2.8 (95% CI, 2.4-3.1) (venous thromboembolism x anticoagulation use interaction on idiopathic interstitial pneumonia outcome: P = 1.5 x 10(-10)). In the general population, ever-diagnosed venous thromboembolism was associated with idiopathic interstitial pneumonia, particularly among those never treated with anticoagulants.


Fundamental strategies for skin care.
The Agency for Health Care Policy and Research (AHCPR) identified the need to develop clinical practice guidelines for the prevention and treatment of pressure ulcers, a segment of wound and skin diagnoses. The skin is the first line of defense in protecting the body from constant changes in the environment. Far too often, the attention needed for the skin is only realized after the integrity has been disrupted. This article examines the anatomy and physiology of the skin, factors that affect the skin (aging, bathing, cleansers, dryness, friction, lotions, moisturizers, nutrition, soaps and shearing forces), indications and contraindications of topical skin care products, and AHCPR's recommended interventions. These elements are integral to clinicians in proactively developing sound skin care prevention strategies. Yet without a thorough understanding of all components, the skin's integrity remains at risk.


Ursodeoxycholic Acid Response Is Associated With Reduced Mortality in Primary Biliary Cholangitis With Compensated Cirrhosis.
Patients with cirrhosis and men have been under-represented in most studies examining the clinical benefit of response to ursodeoxycholic acid (UDCA) in primary biliary cholangitis (PBC). The aim of this study was to study the association of UDCA response and liver-related death or transplantation, hepatic decompensation, and hepatocellular carcinoma (HCC) in patients with PBC cirrhosis. We conducted a retrospective cohort study of veterans, predominantly men, with PBC and compensated cirrhosis to assess the association of UDCA response with the development of all-cause and liver-related mortality or transplantation, hepatic decompensation, and HCC using competing risk time-updating Cox proportional hazards models. We identified 501 subjects with PBC and compensated cirrhosis, including 287 UDCA responders (1,692.8 patient-years [PY] of follow-up) and 214 partial responders (838.9 PY of follow-up). The unadjusted rates of hepatic decompensation (3.8 vs 7.9 per 100 PY, P < 0.0001) and liver-related death or transplantation (3.7 vs 6.2 per 100 PY, P < 0.0001) were lower in UDCA responders compared with partial responders. UDCA response was associated with a lower risk of hepatic decompensation (subhazard ratio [sHR] 0.54, 95% confidence interval [CI] 0.31-0.95, P = 0.03), death from any cause or transplantation (adjusted hazard ratio 0.49, 95% CI 0.33-0.72, P = 0.0002), and liver-related death or transplantation (sHR 0.40, 95% CI 0.24-0.67, P = 0.0004), but not HCC (sHR 0.39, 95% CI 0.60-2.55, P = 0.32). In a sensitivity analysis, the presence of portal hypertension was associated with the highest UDCA-associated effect. UDCA response is associated with a reduction in decompensation, all-cause, and liver-related death or transplantation in a cohort of predominantly male patients with cirrhosis, with the highest benefit in patients with portal hypertension.


Organoids in the Clinic: A Systematic Review of Outcomes.
Research on organoids has undergone significant advances during the last decade. However, outcomes from the use of organoids in clinical trials have not yet been documented. Therefore, there is an urgent need to assess the reporting of clinically relevant outcomes from organoid research in the scientific literature. This article presents a systematic review and appraisal of the published literature in line with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, together with a synopsis of recent relevant reviews. Surprisingly, no randomized controlled trials have reported clinical outcomes with any types of organoids. We found very few ongoing and registered studies that may provide clinically relevant results within this decade. Our screening and interpretation of the literature, including review articles, indicate a focus on technical and preclinical aspects of organoid research. This is the first systematic review of clinical trials involving organoids. Few clinical studies are planned or already underway, and, so far, no high-quality evidence relating to the clinical outcomes of organoid research has been published. The many promises of organoid research still need to be translated from bench to bed.


Obstacles to utilization of prepaid mental health care.
The coverage of out-of-hospital expenses for mental health care through prepayment and insurance programs has been a major advance in removing the economic barriers to early detection, diagnosis, and treatment of mental disorders. However, obstacles to obtaining treatment block many eligible people from receiving help. The authors conducted a study among members of the United Auto Workers union to identify these roadblocks. Among the obstacles were differences in perceptions of need for treatment and in attitudes toward treatment, and lack of awareness of eligibility for benefits among three key groups - consumers, referral agents, and providers. The authors urge psychiatry to help in removing these obstacles.


Challenges in developing a consensus definition of neonatal sepsis.
Sepsis remains a leading cause of morbidity and mortality in the neonatal population, and at present, there is no unified definition of neonatal sepsis. Existing consensus sepsis definitions within paediatrics are not suited for use in the NICU and do not address sepsis in the premature population. Many neonatal research and surveillance networks have criteria for the definition of sepsis within their publications though these vary greatly and there is typically a heavy emphasis on microbiological culture. The concept of organ dysfunction as a diagnostic criterion for sepsis is rarely considered in neonatal literature, and it remains unclear how to most accurately screen neonates for organ dysfunction. Accurately defining and screening for sepsis is important for clinical management, health service design and future research. The progress made by the Sepsis-3 group provides a roadmap of how definitions and screening criteria may be developed. Similar initiatives in neonatology are likely to be more challenging and would need to account for the unique presentation of sepsis in term and premature neonates. The outputs of similar consensus work within neonatology should be twofold: a validated definition of neonatal sepsis and screening criteria to identify at-risk patients earlier in their clinical course. IMPACT: There is currently no consensus definition of neonatal sepsis and the definitions that are currently in use are varied.A consensus definition of neonatal sepsis would benefit clinicians, patients and researchers.Recent progress in adults with publication of Sepsis-3 provides guidance on how a consensus definition and screening criteria for sepsis could be produced in neonatology.We discuss common themes and potential shortcomings in sepsis definitions within neonatology.We highlight the need for a consensus definition of neonatal sepsis and the challenges that this task poses.


[Do left ventricular mass, diameters and long-axis systolic function differ according to angiotensin converting enzyme genotypes?].
Angiotensin converting enzyme (ACE) is a key enzyme in angiotensin II production which causes myocardial hypertrophy and hyperplasia. In this study we aimed to investigate the relation between ACE I/D gene polymorphism and left ventricular mass (LVM), dimensions and systolic functions calculated by mitral annular motion (MAM) in young healthy male subjects. Complete echocardiographic examination was performed in 49 male healthy subjects (mean age 22.9+/-2.1 years) consisting of 18 ACE DD, 18 ACE DI and 13 ACE II genotypes. We calculated LVM and mass index (LVMI) by M-Mode echocardiography. The systolic MAM was recorded at 4 sites (septal, lateral, anterior, and posterior) by M-mode echocardiography and the MAM-ejection fraction (EF) was calculated from above four sites. Ejection fraction was also calculated by Simpson's method. There was no significant difference among the three genotypes according to age, body mass index, systolic and diastolic blood pressure and heart rate. Interventricular septum (IVS) and left ventricular posterior wall (LVPW) diastolic thickness, LVM and LVMI were found significantly different among 3 ACE genotypes. Those measurements were higher in DD genotype in comparison to the DI and II genotypes. There was no significant difference among the three genotypes according to EF-MAM and EF by Simpson's method. In young healthy male subjects having ACE DD genotype, even though LVM and LVMI were within normal limits, their measurements were found to be higher than in subjects with ACE DI and II genotypes respectively. There was no difference among the three genotypes according to left ventricular systolic functions.


Lipophosphoglycans from dermotropic Leishmania infantum are more pro-inflammatory than those from viscerotropic strains.
Although Leishmania infantum is well-known as the aethiological agent of visceral leishmaniasis (VL), in some Central American countries it may cause atypical non-ulcerated cutaneous leishmaniasis (NUCL). However, the mechanisms favoring its establishment in the skin are still unknown. Lipophosphoglycan (LPG) is the major Leishmania multivirulence factor involved in parasite-host interaction. In the case of viscerotropic L. infantum, it causes an immunosuppression during the interaction with macrophages. Here, we investigated the biochemical and functional roles of LPGs from four dermotropic L. infantum strains from Honduras during in vitro interaction with murine macrophages. LPGs were extracted, purified and their repeat units analysed. They did not have side chains consisting of Gal(β1,4)Man(α1)-PO4 common to all LPGs. Peritoneal macrophages from BALB/c and C57BL/6 were exposed to LPG for nitric oxide (NO) and cytokine (TNF-α and, IL-6) production. LPGs from dermotropic strains from Honduras triggered higher NO and cytokine levels compared to those from viscerotropic strains. In conclusion, LPGs from dermotropic strains are devoid of side-chains and exhibit high pro-inflammatory activity.


Status of certain additional over-the-counter drug category II and III active ingredients. Final rule.
The Food and Drug Administration (FDA) is issuing a final rule stating that a certain ingredient in over-the-counter (OTC) drug products is not generally recognized as safe and effective or is misbranded. FDA is issuing this final rule after considering the reports and recommendations of various OTC drug advisory review panels and public comments on proposed agency regulations. This final rule addresses the ingredient octoxynol 9, considered in the rulemaking for OTC vaginal contraceptive drug products. Based on the failure of interested parties to submit new data or information to FDA under the proposed regulation, the agency has determined that the presence of this active ingredient in an OTC drug product would result in that drug product not being generally recognized as safe and effective for its intended use or would result in misbranding. This final rule is part of FDA's ongoing OTC drug product review.


Attractive male sticklebacks carry more oxidative DNA damage in the soma and germline.
Trade-offs between the expression of sexual signals and the maintenance of somatic and germline tissues are expected when these depend upon the same resources. Despite the importance of sperm DNA integrity, its trade-off with sexual signalling has rarely been explored. We experimentally tested the trade-off between carotenoid-based sexual coloration and oxidative DNA damage in skeletal muscle, testis and sperm by manipulating reproductive schedule (early vs. late onset of breeding) in male three-spined sticklebacks. Oxidative DNA damage was measured as the amount of 8-hydroxy-2-deoxyguanosine in genomic DNA. Irrespective of the experimentally manipulated reproductive schedule, individuals investing more in red coloration showed higher levels of oxidative DNA damage in muscle, testis and sperm during the peak breeding season. Our results show that the expression of red coloration traded off against the level of oxidative DNA damage possibly due to the competing functions of carotenoids as colorants and antioxidants. Thus, female sticklebacks may risk fertility and viability of offspring by choosing redder, more deteriorated partners with decreased sperm DNA integrity. The evolution of sexual signal may be constrained by oxidative DNA damage in the soma and germline.


Tumor Vaccines for Malignant Gliomas.
Despite continued research efforts, glioblastoma multiforme (GBM) remains the deadliest brain tumor. Immunotherapy offers a novel way to treat this disease, the genetic signature of which is not completely elucidated. Additionally, these tumors are known to induce immunosuppression in the surrounding tumor microenvironment via an array of mechanisms, making effective treatment all the more difficult. The immunotherapeutic strategy of using tumor vaccines offers a way to harness the activity of the host immune system to potentially control tumor progression. GBM vaccines can react to a variety of tumor-specific antigens, which can be harvested from the patient's unique pathological condition using selected immunotherapy techniques. This article reviews the rationale behind and development of GBM vaccines, the relevant clinical trials, and the challenges involved in this treatment strategy.


Food Processing and Risk of Crohn's Disease and Ulcerative Colitis: A European Prospective Cohort Study.
Industrial foods have been associated with increased risks of several chronic conditions. We investigated the relationship between the degree of food processing and risks of Crohn's disease (CD) and ulcerative colitis (UC) in the European Prospective Investigation into Cancer and Nutrition cohort. Analyses included 413,590 participants (68.6% women; mean baseline age, 51.7 y) from 8 European countries. Dietary data were collected at baseline from validated country-specific dietary questionnaires. Associations between proportions of unprocessed/minimally processed and ultraprocessed food intake and CD and UC risks were estimated using Cox models to obtain hazard ratios (HRs) and 95% CIs. Models were stratified by center, age, and sex, and adjusted for smoking status, body mass index, physical activity, energy intake, educational level, and alcohol consumption. During a mean follow-up period of 13.2 years, 179 incident cases of CD and 431 incident cases of UC were identified. The risk of CD was lower in people consuming high proportions of unprocessed/minimally processed foods (adjusted HR for the highest vs lowest quartile: 0.57; 95% CI, 0.35-0.93; P trend < .01), particularly fruits and vegetables (adjusted HRs, 0.54; 95% CI, 0.34-0.87 and 0.55; 95% CI, 0.34-0.91, respectively). There was no association between unprocessed/minimally processed food intake and the risk of UC. No association was detected between ultraprocessed food consumption and CD or UC risks. In the European Prospective Investigation into Cancer and Nutrition cohort, consumption of unprocessed/minimally processed foods was associated with a lower risk of CD. No association between UC risk and food processing was found.


Evaluating children's literature as a source for patient education.
Children's literature can be used to educate patients about illness, surgery, and hospitalization. However, nurses must evaluate popular literature for developmental appropriateness and content accuracy. Developmental considerations for selecting appropriate children's literature as patient education material include cognitive abilities, language development, and children's literary preferences. Content accuracy is judged based on current practice. Twenty-three children's books addressing surgery and hospitalization were reviewed by the senior author. A seven-step process was utilized to determine the appropriateness of each piece of children's literature reviewed. Based on the review, four common content inaccuracies were identified: (a) the depiction of nurses and their roles, (b) parental visitation policies, (c) preoperative preparation requirements, and (d) the depiction of inpatient surgeries that are currently performed on an outpatient basis. Only four of the 23 books reviewed were recommended to prepare children for surgery in the authors' community. The process of reviewing a book is demonstrated using the example of Curious George Goes to the Hospital.


The genome of Oryctes rhinoceros nudivirus provides novel insight into the evolution of nuclear arthropod-specific large circular double-stranded DNA viruses.
The Oryctes rhinoceros nudivirus (OrNV) is a dsDNA virus with enveloped, rod-shaped virions. Its genome is 127,615 bp in size and contains 139 predicted protein-coding open reading frames (ORFs). In-depth genome sequence comparisons revealed a varying number of shared gene homologues, not only with other nudiviruses (NVs) and baculoviruses, but also with other arthropod-specific large dsDNA viruses, including the so-called Monodon baculovirus (MBV), the salivary gland hypertrophy viruses (SGHVs) and white spot syndrome virus (WSSV). Nudivirus genomes contain 20 baculovirus core gene homologues associated with transcription (p47, lef-8, lef-9, lef-4, vlf-1, and lef-5), replication (dnapol and helicase), virus structure (p74, pif-1, pif-2, pif-3, 19kda/pif-4, odv-e56/pif-5, vp91, vp39, and 38K), and unknown functions (ac68, ac81, and p33). Most strikingly, a set of homologous genes involved in peroral infection (p74, pif-1, pif-2, and pif-3) are common to baculoviruses, nudiviruses, SGHVs, and WSSV indicating an ancestral mode of infection in these highly diverged viruses. A gene similar to polyhedrin/granulin encoding the baculovirus occlusion body protein was identified in non-occluded NVs and in Musca domestica SGHV evoking the question of the evolutionary origin of the baculovirus polyhedrin/granulin gene. Based on gene homologies, we further propose that the shrimp MBV is an occluded member of the nudiviruses. We conclude that baculoviruses, NVs and the shrimp MBV, the SGHVs and WSSV share the significant number of conserved genetic functions, which may point to a common ancestry of these viruses.


The metabolism of proline as microenvironmental stress substrate.
Proline, a unique proteogenic secondary amino acid, has its own metabolic system with special features. Recent findings defining the regulation of this system led us to propose that proline is a stress substrate in the microenvironment of inflammation and tumorigenesis. The criteria for proline as a stress substrate are: 1) the enzymes utilizing proline respond to stress signaling; 2) there is a large, mobilizable pool of proline; and 3) the metabolism of proline serves special stress functions. Studies show that the proline-utilizing enzyme, proline oxidase (POX)/proline dehydrogenase (PRODH), responds to genotoxic, inflammatory, and nutrient stress. Proline as substrate is stored as collagen in extracellular matrix, connective tissue, and bone and it is rapidly released from this reservoir by the sequential action of matrix metalloproteinases, peptidases, and prolidase. Special functions include the use of proline by POX/PRODH to generate superoxide radicals that initiate apoptosis by intrinsic and extrinsic pathways. Under conditions of nutrient stress, proline is an energy source. It provides carbons for the tricarboxylic acid cycle and also participates in the proline cycle. The latter, catalyzed by mitochondrial POX and cytosolic pyrroline-5-carboxylate reductase, shuttles reducing potential from the pentose phosphate pathway into mitochondria to generate ATP and oxidizing potential to activate the cytosolic pentose phosphate pathway.


Fimbriae from the oral anaerobe Bacteroides gingivalis: physical, chemical, and immunological properties.
Circular dichroism spectra indicated the predominance of beta-sheet structure in Bacteroides gingivalis fimbriae regardless of the presence of sodium dodecyl sulfate. By using a computer program, the alpha-helix, beta-sheet, and beta-turn contents and the remainder were estimated to be 0, 55, 18, and 27%, respectively, judging from the circular dichroism spectra of the fimbriae. Heating for 5 min at 100 degrees C in sodium dodecyl sulfate was necessary to denature the fimbriae into their constituent protein (fimbrilin) monomers with a reduced content of beta-sheet structure. The amino-terminal amino acid sequence of the fimbrilin was different from partial or complete amino acid sequences of fimbrilins so far determined from Bacteroides nodosus, which falls into the same nonfermentative species of the genus Bacteroides as B. gingivalis, and from various other bacteria. Fimbrilin monomers had an isoelectric point of 6.0. Examination of antibodies against fimbriae and sodium dodecyl sulfate-denatured fimbrilin by enzyme-linked immunosorbent assay reinforced a previous notion (F. Yoshimura, K. Takahashi, Y. Nodasaka, and T. Suzuki, J. Bacteriol. 160:949-957, 1984) that different sets of antigenic determinants seemed to be exposed on their surfaces.


Sjögren's syndrome, cavitating lung disease and high sustained levels of antibodies to serine proteinase 3.
A case is described involving Sjögren's syndrome, high sustained levels of antibodies to serine proteinase 3, and cavitating lung disease. Possible diagnoses accounting for this unusual combination include a novel association of Sjögren's syndrome and Wegener's granulomatosis (suggested by the high and sustained levels of antibodies to serine proteinase 3) or a rare presentation of bronchiolitis obliterans organising pneumonia. Identification of the true nature of the patients illness facilitated more active management and a swift resolution of the clinical problem.


[Perinatal health in Rotterdam, the Netherlands--experiences after 2 years of 'Ready for a baby'].
The primary goal of the Rotterdam-based health programme 'Ready for a baby' is an improvement in perinatal results via improved, risk-directed care shared across disciplines. After 2 years we conclude that many women who are or who will become pregnant lack knowledge about perinatal risk factors. They are, however, positive about preconception care. High-risk groups can best be accessed via group-based education; results of individual preconceptual care consultations are still limited. We developed and tested a new risk-screening instrument to screen pregnant women during the first antenatal consultation. We directed more attention to non-medical risks, which are often poverty-related. The experiences from our Rotterdam-based programme provide the incentive to experiment with programme-based preconception care, new risk selection during pregnancy and accessing high-risk groups in other communities in the Netherlands.


Draft Genome Sequence of the Urinary Catheter Isolate Enterobacter ludwigii CEB04 with High Biofilm Forming Capacity.
: Enterobacter ludwigii is a fermentative Gram-negative environmental species and accidental human pathogen that belongs to the Enterobacter cloacae complex with the general characteristics of the genus Enterobacter. The clinical isolate E. ludwigii CEB04 was derived from a urinary tract catheter of an individual not suffering from catheter-associated urinary tract infection. The draft genome sequence of the high biofilm forming E. ludwigii CEB04 was determined by PacBio sequencing. The chromosome of E. ludwigii CEB04 is comprised of one contig of 4,892,375 bps containing 4596 predicted protein-coding genes and 120 noncoding RNAs. E. ludwigii CEB04 harbors several antimicrobial resistance markers and has an extended cyclic-di-GMP signaling network compared to Escherichia coli K-12.


Familial visceral myopathy: the role of surgery.
Familial visceral myopathy is a form of chronic intestinal pseudoobstruction in which fibrosis of the smooth muscle of the alimentary tract causes protean symptoms of disordered visceral motility. Based on our 8 year observation of six members of one family with familial visceral myopathy and review of the few well-documented case reports of this disease, we have confirmed a role for surgeons in tissue diagnosis, case identification, and palliation of incapacitating symptoms of familial visceral myopathy after obtaining dynamic radiographic assessment of the entire alimentary canal. Because the duodenum and colon are the usual sources of major dysmotility symptoms, patients incapacitated by duodenal or colonic pseudoobstruction may benefit from bypass or resective operations.


A road map for the development of community systems (CoSy) biology.
Microbial interactions are essential for all global geochemical cycles and have an important role in human health and disease. Although we possess general knowledge about the major processes within a microbial community, we are presently unable to decipher what role individual microorganisms have and how their individual actions influence others in the community. We also have limited knowledge with which to predict the effects of microbial interactions and community composition on the environment and vice versa. In this Opinion article, we describe how community systems (CoSy) biology will enable us to decode these complex relationships and will therefore improve our understanding of individual members of the community and the modes of interactions in which they engage.


Connective tissue growth factor is up-regulated in the diabetic retina: amelioration by angiotensin-converting enzyme inhibition.
Connective tissue growth factor (CTGF) has been postulated to have prosclerotic and angiogenic properties. The aim of this present study was to characterize retinal CTGF expression in the absence and presence of diabetes and in the context of treatment with the angiotensin-converting enzyme (ACE) inhibitor, perindopril. Retinas were obtained from control, diabetic, and diabetic plus perindopril-treated (3 mg/d) rats. CTGF gene expression was quantitated by RT-PCR and localized by in situ hybridization. CTGF protein expression was analyzed by Western blotting and localized by immunohistochemistry. Diabetes was associated with a greater than 2-fold increase in CTGF mRNA levels, which was attenuated by perindopril treatment. CTGF immunoreactivity was increased almost 2-fold in diabetes and was ameliorated by the ACE inhibitor perindopril. By in situ hybridization and immunohistochemistry, the major site of CTGF gene expression in the retina of diabetic rats was the ganglion cell layer. Based on the known in vivo effects of CTGF, it is postulated that this growth factor plays a pivotal role in mediating diabetes-associated retinal pathology. Furthermore, the protective effects of ACE inhibitors on retinal pathology may partly be mediated via effects on retinal CTGF expression.


Effects of rosiglitazone on inflammation in Otsuka long-evans Tokushima Fatty rats.
Inflammation plays a role in the response to metabolic stress in type 2 diabetes. However, the effects of rosiglitazone on inflammation of skeletal muscle have not been fully examined in type 2 diabetes. We investigated the effects of the insulin-sensitizing anti-diabetic agent, rosiglitazone, on the progression of skeletal muscle inflammation in Otsuka Long-Evans Tokushima Fatty (OLETF) type 2 diabetic rats. We examined the expression of serologic markers (serum glucose, insulin and free fatty acid) and inflammatory cytokines (tumor-necrosis factor-alpha, interleukin [IL]-1beta and IL-6) in OLETF rats from early to advanced diabetic stage (from 28 to 40 weeks of age). Serum glucose and insulin concentrations were significantly decreased in rosiglitazone-treated OLETF rats compared to untreated OLETF rats. Rosiglitazone treatment significantly decreased the concentrations of serum inflammatory cytokines from 28 to 40 weeks of age. The mRNA expression of various cytokines in skeletal muscle was reduced in rosiglitazone-treated OLETF rats compared with untreated OLETF rats. Furthermore, rosiglitazone treatment resulted in the downregulation of ERK1/2 phosphorylation and NF-kappaB expression in the skeletal muscle of OLETF rats. These results suggest that rosiglitazone may improve insulin sensitivity with its anti-inflammatory effects on skeletal muscle.


[Enzootic bovine leukosis--a retrovirus disease].
After a short historical introduction and a description of some of the properties of the bovine leucosis (leukemia) virus (BLV) the natural pathways of transmission, for which most likely intact infected cells are a prerequisite, are described and the role of possible vectors mentioned. The iatrogenic transmission by needles, dehorning and perhaps even rectal examinations is discussed. Semen, embryo transfer, saliva, urine, faeces and breath do obviously not play any role in contrast to nasal secretions containing viable cells. It is pointed out that it is not inevitable to eliminate all seropositive cattle from the premises during an eradication programme provided some additional tests are carried out: determination of the white blood picture, the p 24 status and perhaps even proof of antigen production in short-term buffy coat cultures. Offspring of seropositive cattle requires some special attention and careful testing at certain intervals. The report according to which BVD/MD virus-infected cattle are superinfected with BLV leading to rather difficult situations is cited.


Resolving pathways of interaction of mipafox and a sarin analog with human acetylcholinesterase by kinetics, mass spectrometry and molecular modeling approaches.
The hydroxyl oxygen of the catalytic triad serine in the active center of serine hydrolase acetylcholinesterase (AChE) attacks organophosphorus compounds (OPs) at the phosphorus atom to displace the primary leaving group and to form a covalent bond. Inhibited AChE can be reactivated by cleavage of the Ser-phosphorus bond either spontaneously or through a reaction with nucleophilic agents, such as oximes. At the same time, the inhibited AChE adduct can lose part of the molecule by progressive dealkylation over time in a process called aging. Reactivation of the aged enzyme has not yet been demonstrated. Here, our goal was to study oxime reactivation and aging reactions of human AChE inhibited by mipafox or a sarin analog (Flu-MPs, fluorescent methylphosphonate). Progressive reactivation was observed after Flu-MPs inhibition using oxime 2-PAM. However, no reactivation was observed after mipafox inhibition with 2-PAM or the more potent oximes used. A peptide fingerprinted mass spectrometry (MS) method, which clearly distinguished the peptide with the active serine (active center peptide, ACP) of the human AChE adducted with OPs, was developed by MALDI-TOF and MALDI-TOF/TOF. The ACP was detected with a diethyl-phosphorylated adduct after paraoxon inhibition, and with an isopropylmethyl-phosphonylated and a methyl-phosphonylated adduct after Flu-MPs inhibition and subsequent aging. Nevertheless, nonaged nonreactivated complexes were seen after mipafox inhibition and incubation with oximes, where MS data showed an ACP with an NN diisopropyl phosphoryl adduct. The kinetic experiments showed no reactivation of activity. The computational molecular model analysis of the mipafox-inhibited hAChE plots of energy versus distance between the atoms separated by dealkylation showed a high energy demand, thus little aging probability. However, with Flu-MPs and DFP, where aging was observed in our MS data and in previously published crystal structures, the energy demand calculated in modeling was lower and, consequently, aging appeared as a more likely reaction. We document here direct evidence for a phosphorylated hAChE refractory to oxime reactivation, although we observed no aging.


Allogeneic bone marrow transplantation for acute nonlymphocytic leukemia in first remission.
Seventy-three patients with acute nonlymphocytic leukemia in first complete remission (CR) have received allogeneic bone marrow transplantation (BMT) with non-T-lymphocyte-depleted marrow obtained from matched sibling donors. The first 36 patients received a preparative regimen consisting of cyclophosphamide, 60 mg/kg/d (days -6 and -5), and 750 cGy single-dose total-body irradiation (TBI) (day -1). Subsequently, 37 patients received cyclophosphamide 60 mg/kg/d (days -6 and -5), and 165 cGy fractionated TBI administered twice daily for a total dose of 1,320 cGy (days -4, -3, -2, and -1). Survivors have been followed from 9 to 124 months (median, 40 months). The 61% (95% confidence interval [CI], 45% to 77%) projected disease-free survival (DFS) of 41 children less than 18 years old does not differ significantly from the 62% (95% CI, 49% to 73%) projected DFS of 32 adults at 84 months (P = .89). Similarly, the 15% (95% CI, 1% to 29%) projected relapse rate seen in children does not differ from the 9% (95% CI, 0% to 21%) seen in adults (P = .69). Multivariate Cox regression analysis of presenting features demonstrates that a presenting WBC count greater than 20,000/m3 is associated with decreased DFS (P = .01). When compared with other French-American-British (FAB) subtypes, presentation with FAB M4 or M5 morphology is significantly associated with relapse in multivariate analysis (P = .014). Other presenting features such as preparation with single-dose or fractionated TBI, interval from diagnosis to CR or CR to BMT, donor or recipient sex, and donor or recipient cytomegalovirus serology do not correlate independently with either DFS or relapse. When included in the stepwise multivariate analysis of presenting patient features, two posttransplant events, development of grades 2 to 4 acute graft-v-host disease (GVHD) (P less than .03) and development of interstitial pneumonitis (P less than .001), also correlate independently with poor DFS. Allogeneic BMT provides equivalent, prolonged DFS in both children and young adults when performed in first CR and should be considered the therapy of choice for all first CR patients under 45 years of age with a suitable donor. Continued efforts to prevent and treat acute GVHD and pneumonitis as well as efforts designed to prevent relapse in patients presenting with FAB M4 and M5 morphology should further improve outcome.


Comparison between in vitro neutralization titres and in vivo protection against homologous and heterologous challenge induced by vaccines prepared from two serologically distinct variants of foot-and-mouth disease virus, serotype A22.
Guinea-pigs were challenged with homologous or heterologous strains of foot-and-mouth disease virus (FMDV) following vaccination with baby hamster kidney (BHK) monolayer cell-adapted or BHK suspension cell-adapted strains of FMDV serotype A22 Iraq 24/64. The protection afforded by these vaccines was analysed as a function of antigen dose and the in vitro serum virus neutralization titres achieved. The results show that the level of neutralizing antibody induced that afforded 50% protection was similar for both vaccines in homologous or heterologous challenge situations. However, although the dose of antigen required to achieve this titre against homologous virus was similar for the two vaccines, approximately 20-fold more of the suspension cell-adapted virus was required to elicit a protective titre against heterologous challenge compared to the dose of monolayer cell-adapted virus required. A synthetic peptide representing the amino acid sequence 135 to 167 of VP1, which is identical in the A22 Iraq 24/64 variant viruses, was shown to induce protection against both homologous and heterologous virus challenge.


The investment case for preventing NICU-associated infections.
Nosocomial [hospital-associated or neonatal intensive care unit (NICU)-associated] infections occur in as many as 10 to 36% of very low-birth-weight infants cared for in NICUs. To determine the potentially avoidable, incremental costs of care associated with NICU-associated bloodstream infections. This retrospective study included all NICU admissions of infants weighing 401 to 1500 g at birth in the greater Cincinnati region from January 1, 2005, through December 31, 2007. Nonphysician costs of care were compared between infants who developed at least one bacterial bloodstream infection prior to NICU discharge or death and infants who did not. Costs were adjusted for clinical and demographic characteristics that are present in the first 3 days of life and are known associates of infection. Among 900 study infants with no congenital anomaly and no major surgery, 82 (9.1%) developed at least one bacterial bloodstream infection. On average, the cost of NICU care was $16,800 greater per infant who experienced NICU-associated bloodstream infection. Potentially avoidable costs of care associated with bloodstream infection can be used to justify investments in the reliable implementation of evidence-based interventions designed to prevent these infections.


Mothers' insightfulness regarding their infants' internal experience: relations with maternal sensitivity and infant attachment.
This study examined the associations among mothers' insightfulness into their infants' internal experience, mothers' sensitivity to their infants' signals, and infants' security of attachment to their mothers. The insightfulness of 129 mothers of 12-month-old infants was assessed by showing mothers 3 videotaped segments of observations of their infants and themselves and interviewing them regarding their infants' and their own thoughts and feelings. Interviews were classified into 1 insightful and 3 noninsightful categories. Mothers' sensitivity was assessed during play sessions at home and at the laboratory, and infant-mother attachment was assessed with the Strange Situation. Mothers classified as positively insightful were rated as more sensitive and were more likely to have securely attached children than were mothers not classified as positively insightful. Insightfulness also accounted for variance in attachment beyond the variance explained by maternal sensitivity. These findings add an important dimension to research on caregiving, suggesting that mothers' seeking of explanations for the motives underlying their infants' behavior is related to both maternal sensitivity and infant attachment.


Effects of fixed combination of lisinopril plus hydrochlorothiazide on regression of left ventricular hypertrophy in patients with essential hypertension: an opened, multi-centre, prospective clinical trial.
The aim of this trial was to examine the effects of antihypertensive fixed combination of lisinopril plus hydrochlorothiazide (Lopril H, Bosnalijek dd, Bosnia and Herzegovina) on regression of left ventricular hypertrophy in patients with essential arterial hypertension. We included 297 patients in our trial, aged 54.65+/-9.6 years, with treated or untreated hypertension and with high risk of cardiac events, in an opened trial of therapy based on lisinopril plus hydrochlorothiazide. Patients from five European countries were followed up for a period of 12 weeks. Duration of treatment was 12 weeks. We adjusted daily doses of lisinopril plus hydrochlorothiazide after every clinical examination and recorded adverse effects of drugs. In the beginning and after 12 weeks of treatment, 277 patients (93.2%) underwent 2-dimensional echocardiography and there were 186 patients evaluated for efficacy of treatment on left ventricular hypertrophy (LVH). We recorded a regression of index mass LVH (168.56 vs 161.51 g/m2, P<0.0001), and regression was something more in women vs men. We recorded average reduction of left ventricular mass index for patients with LVH (N=186) by 7.05 g/m2 (4.18%) in all patients, by 6.73 g/m2 (3.93%) in men and 7.27 g/m2 (4,37%) in women. The proportion of patients who attained a regression of left ventricular mass tended to be greater in men (54.55% vs 53.21%). This research has proved regression of LVH in more than 53% patients after using fixed combination of lisinopril plus hydrochlorothiazide.


Analysis of material selection on dielectric metasurface performance.
Dielectric metasurfaces are ultra-thin devices that can shape optical wavefronts with extreme control. While an assortment of materials possessing a wide range of dielectric constants have been proposed and implemented, the minimum dielectric contrast required for metasurfaces to achieve high-efficiency performance, for a given device function and feature size constraint, is unclear. In this Article, we examine the impact of dielectric material selection on metasurface efficiency at optical frequencies. As a model system, we design transmissive, single-layer periodic metasurfaces (i.e., metagratings) using topology optimization, and we sweep device thickness and light deflection angle for differing material types. We find that for modest deflection angles below 40 degrees, materials with relatively low dielectric constants near 4 can be used to produce metagratings with efficiencies over 80%. However, ultra-high-efficiency devices designed for large deflection angles and multiple functions require materials with high dielectric constants comparable to silicon. We also identify, for all materials, a minimum device thickness required for optimal metagrating performance that scales inversely with dielectric constant. Our work presents materials selection guidelines for high-performance metasurfaces operating at visible and infrared wavelengths.


Spondylolysis 2019 update.
Provide a comprehensive overview of lumbar spondylolysis, a frequent cause of lower back pain in children and adolescents, from evaluation to management. With the surge of structured sports participation in the pediatric population, spondylolysis is a common ailment that afflicts many young athletes due to rigorous competition that taxes the growing spine with repetitive extension and rotation. Spondylolysis is a fracture through the pars interarticularis. When a child presents with lower back pain, spondylolysis should be at the top of the differential. A thorough history and physical examination are essential. In addition, radiographs of the lumbar spine, anterior posterior and lateral views, MRI and, selectively, computed tomography are useful adjuncts to uncover a pars injury. Timely diagnosis facilitates early treatment which includes rest, optimizing bone health, brace treatment, and physical therapy. If delayed or untreated, spondylolysis may result in nonunion or pars defect. If symptomatic, patients may need surgery to fuse the lumbar facet joints or repair the par interarticularis.


Wnt/β‑catenin inhibition reverses multidrug resistance in pediatric acute lymphoblastic leukemia.
Although ~80% of newly diagnosed pediatric patients with acute lymphoblastic leukemia (ALL) become disease‑free following appropriate treatment, relapses frequently occur, with dismal prognosis. Therefore, it is urgent to develop novel therapeutic modalities. Resistance to chemotherapy is a major obstacle for the treatment of relapsed ALL. It has been indicated that Wnt pathway is potentially associated with leukemia recurrence. In the current study, a vincristine (VCR)‑resistant variant of the human ALL cell line BALL‑1 (BALL‑1/VCR) that also had relatively specific resistance to both doxorubicin and etoposide was generated. Over‑activation of the Wnt/β‑catenin signaling pathway was observed in BALL‑1/VCR cells, whereas Dickkopf‑related protein 1 selectively suppressed the Wnt signaling pathway and sensitized the response of BALL‑1/VCR to anticancer agents. In addition, prednisolone exposure in combination with Wnt inhibition restored chemo‑sensitivity in relapsed ALL blasts. Since the resistance of BALL‑1/VCR cells is potentially attributed to the overexpression of MDR‑associated protein 1 (MRP1), the development of drug resistance in relapsed ALL may associated with the overexpression of MRP1 and P‑glycoprotein. The results of this study demonstrated that, as a potential candidate to mimic relapsed ALL, BALL‑1/VCR could be used in further research, while Wnt‑inhibition may become a promising therapeutic approach for treating ALL.


Improving the measurement accuracy of the effort-reward imbalance scales.
The effort-reward imbalance (ERI) scale items are answered in a two-step process, but the justification is questioned for the formulation of summary measure by combining information rated in two steps. To examine whether the basic prerequisites of the ERI scales are empirically satisfied and to seek ways to improve the rating procedure. A polytomous item response theory (IRT) model was applied to the responses of 20,256 workers who completed the ERI scales. To determine the most appropriate statistical justification, three alternative scoring algorithms were compared with regard to the test properties revealed by the IRT analyses and efficiencies of screening performance and criterion validity against depressive symptomatology. The rated raw-score units did not reflect the hypothesized order of lowest stress levels to highest stress levels. Exchanging or collapsing the lowest two categories of a Likert scaled item, where data of different quality are combined, solved this problem, thereby making the test content more appropriate. The modified rating improved the efficiencies of screening performance and the correlation of the stress summary measures against health criterion, i.e., depression. An avoidable measurement error exists in the current ERI scales. Modifying the rating procedure can improve the measurement accuracy.


Retinoic acid loaded with chitosan nanoparticles improves spermatogenesis in scrotal hyperthermia in mice.
High temperatures can trigger cellular oxidative stress and disrupt spermatogenesis, potentially leading to male infertility. We investigated the effects of retinoic acid (RA), chitosan nanoparticles (CHNPs), and retinoic acid loaded with chitosan nanoparticles (RACHNPs) on spermatogenesis in mice induced by scrotal hyperthermia (Hyp). Thirty mice (weighing 25 to 30 g) were divided into five experimental groups of six mice each. The groups were as follows: control, Hyp induced by a water bath (43 °C for 30 minutes/day for 5 weeks), Hyp+RA (2 mg/kg/day), Hyp+CHNPs (2 mg/kg/72 hours), and Hyp+RACHNPs (4 mg/kg/72 hours). The mice were treated for 35 days. After the experimental treatments, the animals were euthanized. Sperm samples were collected for analysis of sperm parameters, and blood serum was isolated for testosterone measurement. Testis samples were also collected for histopathology assessment, reactive oxygen species (ROS) evaluation, and RNA extraction, which was done to compare the expression levels of the bax, bcl2, p53, Fas, and FasL genes among groups. Additionally, immunohistochemical staining was performed. Treatment with RACHNPs significantly increased stereological parameters such as testicular volume, seminiferous tubule length, and testicular cell count. Additionally, it increased testosterone concentration and improved sperm parameters. We observed significant decreases in ROS production and caspase-3 immunostaining in the RACHNP group. Moreover, the expression levels of bax, p53, Fas, and FasL significantly decreased in the groups treated with RACHNPs and RA. RACHNPs can be considered a potent antioxidative and antiapoptotic agent for therapeutic strategies in reproductive and regenerative medicine.


Prognosis and treatment of paraquat poisoning: a review of 28 cases.
Paraquat poisoning is very severe. When it is ingested, this herbicide may be responsible for causative lesions of the digestive tract, cytolytic hepatitis, renal tubular necrosis, circulatory failure, and/or pulmonary fibrosis. Since a very low dose (as little as one mouthful) is potentially lethal, it is important to understand why 11 of our 28 patients who entered our department for paraquat poisoning survived. The main prognostic factors appear to be the following: Route of administration. Of four patients who had inhaled paraquat aerosols and/or contaminated their skin with the herbicide, all survived. Ingested amount. Above 50 mg/kg, patients died of circulatory failure within 72 h; between 35 and 50 mg/kg, a progressive pulmonary fibrosis occurred. Delay between ingestion and the last meal. Paraquat is adsorbed and neutralized by foodstuffs. Caustic gastric lesions revealed by early endoscopic examination. The occurrence of an organic renal failure. The plasma paraquat concentrations within the first 24 h. Patients whose plasma concentrations do not exceed 2.0, 0.6, 0.3, 0.16, and 0.1 mg/L at 4, 6, 10, 16, and 24 h, respectively, are likely to survive. The different treatments that have been tested (fuller's earth, forced diarrhea, furosemide, hemodialysis, hemoperfusion, artificial ventilation with hypoxic breathing mixtures) did not modify the initial prognosis. The 11 survivals are only linked to the circumstances of the poisonings (route of administration, ingested amount, delay between ingestion and the last meal, etc.). The treatments did not modify the outcome.


[Immunohistochemical and clinicopathological study of thymidylate synthase, dihydropyrimidine dehydrogenase, and orotate phosphoribosyl transferase expression in oral cancer patients responding to UFT].
Thymidylate synthase(TS), dihydropyrimidine dehydrogenase(DPD), and orotate phosphoribosyl transferase(OPRT)are initial key enzymes in the 5-fluorouracil(5-FU)metabolic pathway. In this study, we investigated clinicopathological and immunohistochemical expressions of TS, DPD, and OPRT in oral cancer patients who showed a complete response(CR)to UFT. We also evaluated patients showing a partial response(PR)and stable disease(SD)following UFT. The numbers of CR, PR, and SD cases were 3, 5, and 10, respectively. Pathologically, all CR and PR cases were the well-differentiated type, and 5 out of 10 SD cases were of the moderately or poorly-differentiated type. Three out of the 5 cases of moderately or poorlydifferentiated type were DPD-negative. Most cases of CR and PR were DPD-positive. OPRT expression showed no difference with the UFT response. We suggest that UFT affects high DPD patients with the well-differentiated type, but may not influence low DPD patients with the moderately or poorly-differentiated type.


Phytic acid synthesis and vacuolar accumulation in suspension-cultured cells of Catharanthus roseus induced by high concentration of inorganic phosphate and cations.
We have established a new system for studying phytic acid, myo-inositol hexakisphosphate (InsP(6)) synthesis in suspension-cultured cells of Catharanthus. InsP(6) and other intermediates of myo-inositol (Ins) phosphate metabolism were measured using an ion chromatography method. The detection limit for InsP(6) was less than 50 nM, which was sufficient to analyze Ins phosphates in living cells. Synthesis of Ins phosphates was induced by incubation in high inorganic phosphate medium. InsP(6) was mainly accumulated in vacuoles and was enhanced when cells were grown in high concentration of inorganic phosphates with the cations K(+), Ca(2+), or Zn(2+). However, there was a strong tendency for InsP(6) to accumulate in the vacuole in the presence of Ca(2+) and in nonvacuolar compartments when supplied with Zn(2+), possibly due to precipitation of InsP(6) with Zn(2+) in the cytosol. A vesicle transport inhibitor, brefeldin A, stimulated InsP(6) accumulation. The amounts of both Ins(3)P(1) myo-inositol monophosphate synthase, a key enzyme for InsP(6) synthesis, and Ins(1,4,5)P(3) kinase were unrelated to the level of accumulation of InsP(6). The mechanisms for InsP(6) synthesis and localization into vacuoles in plant cells are discussed.


Verbascoside promotes apoptosis by regulating HIPK2-p53 signaling in human colorectal cancer.
We investigated the role of the HIPK2-p53 signaling pathway in tumorigenesis and resistance to the drug Verbascoside (VB) in colorectal cancer (CRC), using in vivo and in vitro experiments. Primary human CRC samples and normal intestinal tissues from patients were analyzed for HIPK2 expression by immunohistochemistry (IHC) and its expression was correlated against patients' clinicopathological characteristics. Human CRC HCT-116 cells were implanted in BALB/c nude mice; mice with xenografted tumors were randomly administrated vehicle (control), 20, 40, or 80 mg/mL VB, or 1 mg/mL fluorouracil (5-FU). HIPK2, p53, Bax, and Bcl-2 expression in these tumors were determined by IHC. In vitro effects of VB on CRC cell proliferation and apoptosis were measured by CCK-8 assay and flow cytometry; HIPK2, p53, p-p53, Bax, and Bcl-2 were measured by western blot. IHC analysis for 100 human CRC tumor samples and 20 normal intestinal tissues, showed HIPK2 expression to inversely correlate with Dukes stage and depth of invasion in CRC (P<0.05). In vivo, the inhibition rates of 20, 40, and 80 mg/mL VB on CRC xenograft tumor weight were 42.79%, 53.90%, and 60.99%, respectively, and were accompanied by increased expression of HIPK2, p53, and Bax, and decreased Bcl-2 expression in treated tumors. In vitro, VB significantly inhibited proliferation of CRC cell lines HCT-116, HT-29, LoVo, and SW620, in a time- and dose-dependent manner. The apoptosis rates of 25, 50, and 100 μM VB on HCT-116 cells were 10.83±1.28, 11.25±1.54, and 20.19±2.87%, and on HT-29 cells were 18.92±6.12, 21.57±4.05, and 25.14±6.73%, respectively. In summary, VB treatment significantly enhanced the protein expression of pro-apoptotic HIPK2, p53, p-p53, Bax, and decreased anti-apoptotic Bcl-2 expression in CRC cells. HIPK2 protein modulates the phosphorylation status of p53, and levels of Bax and Bcl-2 in CRC. We also found that VB effectively activated the HIPK2-p53 signaling pathway, resulting in increased CRC cell apoptosis.


Diversity gradients and phylogeographic patterns in Santiria trimera (Burseraceae), a widespread African tree typical of mature rainforests.
New insights into the history of the African rainforest can be gathered from the phylogeographic structures of their constituent species, but few studies have been performed in this ecosystem. We studied the phylogeographic structure of Santiria trimera, a primate- and bird-dispersed, dioecious tree typical of mature African rainforests. We sequenced three chloroplast DNA (cpDNA) regions (trnL-F, rbcL, and rpl36-infA-rps8) in 377 individuals from 42 populations. Sequence chromatograms regularly displayed double peaks of unequal heights. Cloning of PCR products and sequencing of outgroup taxa led to assigning the taller peak in ambiguous sequence positions to cpDNA. A total of 14 polymorphic cpDNA sites and 12 haplotypes were detected. Populations from three distinct biogeographic regions, namely, Upper Guinea, Lower Guinea, and the volcanic island of São Tomé, did not share any haplotype, indicating allopatric divergence. In Lower Guinea, Gabonese forests had high diversity mainly from the sympatry of two genetically divergent morphotypes, whereas forests of eastern Cameroon were less diversified. The two haplotypes of the morphotype without stilt roots were distributed north and south of the Ogooué River, suggesting refuges on both sides of the river bed. The divergence between Upper and Lower Guinean rainforests is explained by the discontinuity of forest between those regions throughout most of the Quaternary. The distribution of rare endemic haplotypes concurred with proposed Pleistocene rainforest refuges in west and southwest Cameroon. Overall, phylogeographic structure is consistent with the biogeographic hypotheses largely based on patterns of species diversity.


Influence of left ventricular relaxation on the pressure half time of aortic regurgitation.
The severity of aortic regurgitation can be estimated using pressure half time (PHT) of the aortic regurgitation flow velocity, but the correlation between regurgitant fraction and PHT is weak. To test the hypothesis that the association between PHT and regurgitant fraction is substantially influenced by left ventricular relaxation. In 63 patients with aortic regurgitation, subdivided into a group without (n = 22) and a group with (n = 41) left ventricular hypertrophy, regurgitant fraction was calculated using the difference between right and left ventricular cardiac outputs. Left ventricular relaxation was assessed using the early to late diastolic Doppler tissue velocity ratio of the mitral annulus (E/ADTI), the E/A ratio of mitral inflow (E/AM), and the E deceleration time (E-DT). Left ventricular hypertrophy was assessed using the M mode derived left ventricular mass index. The overall correlation between regurgitant fraction and PHT was weak (r = 0.36, p < 0.005). In patients without left ventricular hypertrophy, there was a significant correlation between regurgitant fraction and PHT (r = 0.62, p < 0.005), but not in patients with left ventricular hypertrophy. In patients with a left ventricular relaxation abnormality (defined as E/ADTI< 1, E/AM< age corrected lower limit, E-DT >/= 220 ms), no associations between regurgitant fraction and PHT were found, whereas in patients without left ventricular relaxation abnormalities, the regurgitant fraction to PHT relations were significant (normal E/AM: r = 0.57, p = 0.02; E-DT< 220 ms: r = 0.50, p < 0.001; E/ADTI < 1: r = 0.57, p = 0.02). Only normal left ventricular relaxation allows a significant decay of PHT with increasing aortic regurgitation severity. In abnormal relaxation, which is usually present in left ventricular hypertrophy, wide variation in prolonged backward left ventricular filling may cause dissociation between the regurgitant fraction and PHT. Thus the PHT method should only be used in the absence of left ventricular relaxation abnormalities.


Counselor attitudes toward the use of naltrexone in substance abuse treatment: a multi-level modeling approach.
Alcohol use disorders (AUDs) continue to be one of the most pervasive and costly of the substance use disorders (SUDs). Despite evidence of clinical effectiveness, adoption of medications for the treatment of AUDs is suboptimal. Low rates of AUD medication adoption have been explained by characteristics of both treatment organizations and individual counselor's attitudes and behaviors. However, few studies have simultaneously examined the impact of organizational-level and counselor-level characteristics on counselor perceptions of EBPs. To address this gap in the literature, we use data from a national sample of 1178 counselors employed in 209 privately funded treatment organizations to examine the effects of organizational and individual counselor characteristics on counselor attitudes toward tablet and injectable naltrexone. Results of hierarchical linear modeling (HLM) show that organizational characteristics (use of tablet/injectable naltrexone in the program, 12-step orientation) were associated with counselor perceptions of naltrexone. Net of organizational characteristics, several counselor level characteristics were associated with attitudes toward tablet and injectable naltrexone including gender, tenure in the field, recovery status, percentage of AUD patients, and receipt of medication-specific training. These findings reveal that counselor receptiveness toward naltrexone is shaped in part by the organizational context in which counselors are embedded.


Knockdown of TFIIS by RNA silencing inhibits cancer cell proliferation and induces apoptosis.
A common element among cancer cells is the presence of improperly controlled transcription. In these cells, the degree of specific activation of some genes is abnormal, and altering the aberrant transcription may therefore directly target cancer. TFIIS is a transcription elongation factor, which directly binds the transcription motor, RNA Polymerase II and allows it to read through various transcription arrest sites. We report on RNA interference of TFIIS, a transcription elongation factor, and its affect on proliferation of cancer cells in culture. RNA interference was performed by transfecting siRNA to specifically knock down TFIIS expression in MCF7, MCF10A, PL45 and A549 cells. Levels of TFIIS expression were determined by the Quantigene method, and relative protein levels of TFIIS, c-myc and p53 were determined by C-ELISA. Induction of apoptosis was determined by an enzymatic Caspase 3/7 assay, as well as a non-enzymatic assay detecting cytoplasmic mono- and oligonucleosomes. A gene array analysis was conducted for effects of TFIIS siRNA on MCF7 and MCF10A cell lines. Knockdown of TFIIS reduced cancer cell proliferation in breast, lung and pancreatic cancer cell lines. More specifically, TFIIS knockdown in the MCF7 breast cancer cell line induced cancer cell death and increased c-myc and p53 expression whereas TFIIS knockdown in the non-cancerous breast cell line MCF10A was less affected. Differential effects of TFIIS knockdown in MCF7 and MCF10A cells included the estrogenic, c-myc and p53 pathways, as observed by C-ELISA and gene array, and were likely involved in MCF7 cell-death. Although transcription is a fundamental process, targeting select core transcription factors may provide for a new and potent avenue for cancer therapeutics. In the present study, knockdown of TFIIS inhibited cancer cell proliferation, suggesting that TFIIS could be studied as a potential cancer target within the transcription machinery.


In vitro antibacterial activity of ME1207, a new oral cephalosporin.
ME1207 is the prodrug of ME1206. Its in vitro antibacterial activity was compared with that of cefteram, cefpodoxime, cefixime, and cefaclor against various clinical isolates. ME1206 was more active than the other cephems tested against staphylococci, streptococci, Morganella morganii, Pseudomonas cepacia, and Flavobacterium meningosepticum and had the most potent activity against Haemophilus influenzae and Neiserria gonorrhoeae. The drug also showed a wide spectrum of activity against other gram-positive and gram-negative bacteria, except methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, Citrobacter freundii, Pseudomonas aeruginosa, Xanthomonas maltophilia, and Alcaligenes xylosoxydans.


Autocrine or paracrine inflammatory actions of corticotropin-releasing hormone in vivo.
Corticotropin-releasing hormone (CRH) functions as a regulator of the hypothalamic-pituitary-adrenal axis and coordinator of the stress response. CRH receptors exist in peripheral sites of the immune system, and CRH promotes several immune functions in vitro. The effect of systemic immunoneutralization of CRH was tested in an experimental model of chemically induced aseptic inflammation in rats. Intraperitoneal administration of rabbit antiserum to CRH caused suppression of both inflammatory exudate volume and cell concentration by approximately 50 to 60 percent. CRH was detected in the inflamed area but not in the systemic circulation. Immunoreactive CRH is therefore produced in peripheral inflammatory sites where, in contrast to its systemic indirect immunosuppressive effects, it acts as an autocrine or paracrine inflammatory cytokine.


Aspirin for prevention of colorectal cancer in the elderly: friend or foe?
Cancer is the leading cause of death among men and women aged 60-79 years. Colorectal cancer is the third most common cancer in males and the second most common in females, with about 0.8 million deaths worldwide per year. Individuals older than 50 years account for 20-50% of colonic adenomas. Several measures have been proposed to decrease colorectal cancer risks, such as an increase in dietary fiber, use of aspirin, and physical activity. Nonsteroidal anti-inflammatory drugs have been proposed as protective agents against the development of colorectal cancer and colorectal adenomas. Aspirin was the first pharmacological agent endorsed by the United States Preventive Services Task Force screening for colorectal cancer chemoprevention. Although studies have shown up to 40% colorectal cancer risk reduction in individuals at average risk, data regarding this benefit are inconsistent. Several recent studies show that prophylactic use of aspirin in elderly subjects may not be beneficial in preventing the occurrence of colorectal cancers. Given the risks associated with aspirin, such as non-fatal and fatal bleeding events, aspirin's role should be redefined, especially in individuals at risk of bleeding. This review provides a discussion of the recent studies on the role of aspirin use in elderly individuals at risk of colorectal cancer.


An examination of multiple classes of rare variants in extended families with bipolar disorder.
Bipolar disorder (BD) is a complex psychiatric condition with high heritability, the genetic architecture of which likely comprises both common variants of small effect and rare variants of higher penetrance, the latter of which are largely unknown. Extended families with high density of illness provide an opportunity to map novel risk genes or consolidate evidence for existing candidates, by identifying genes carrying pathogenic rare variants. We performed whole-exome sequencing (WES) in 15 BD families (117 subjects, of whom 72 were affected), augmented with copy number variant (CNV) microarray data, to examine contributions of multiple classes of rare genetic variants within a familial context. Linkage analysis and haplotype reconstruction using WES-derived genotypes enabled exclusion of false-positive single-nucleotide variants (SNVs), CNV inheritance estimation, de novo variant identification and candidate gene prioritization. We found that rare predicted pathogenic variants shared among ≥3 affected relatives were overrepresented in postsynaptic density (PSD) genes (P = 0.002), with no enrichment in unaffected relatives. Genome-wide burden of likely gene-disruptive variants was no different in affected vs. unaffected relatives (P = 0.24), but correlated significantly with age of onset (P = 0.017), suggesting that a high disruptive variant burden may expedite symptom onset. The number of de novo variants was no different in affected vs. unaffected offspring (P = 0.89). We observed heterogeneity within and between families, with the most likely genetic model involving alleles of modest effect and reduced penetrance: a possible exception being a truncating X-linked mutation in IRS4 within a family-specific linkage peak. Genetic approaches combining WES, CNV and linkage analyses in extended families are promising strategies for gene discovery.


Abdominopelvic actinomycosis mimicking disseminated peritoneal carcinomatosis.
We present a case of a 38-year-old woman who presented with symptoms suggestive of intra-abdominal or pelvic malignancy: marked weight loss, abdominal pain, altered bowel habit, anorexia and fatigue. The findings of multiple peritoneal deposits, adnexal and presacral masses on CT imaging and appearances on diagnostic laparotomy also suggested malignancy. However, the histological analysis was inconsistent with malignancy and revealed an infection with Actinomyces israelii. The patient started a course of intravenous antibiotics and complete resolution is expected. An intrauterine contraceptive device was identified as the likely source of the infection.


Development of a Pediatric Cardiac Mechanical Support Program.
The development of a pediatric cardiac support program is a complex, multidisciplinary project. This study describes the University of Iowa Congenital Heart Program's experience from its inception to the present. In, we examine those specific factors that have led to substantial improvements in the program, additionally identifying where further gains can be made. We retrospectively reviewed all pediatric patients who received mechanical cardiac support at the University of Iowa from the inception of the program in 1991. In total, 29 patients received mechanical support between December 1991 and December 2015 and are included in the study. Twelve patients received continuous flow devices and 17 patients received pulsatile flow devices. Median age at implant was 12.8 years (range 0.1-18.2 years). Median weight at implant was 40.5 kg (3.2-123.4 kg). Factors examined included: operating room (OR) time, intensive care unit and hospital length of stay, intubation days, blood product usage, pre- and post-operative bilirubin, creatinine, natriuretic peptide B (NPPB), and device implanted. Categorical and continuous variables were compared using Chi-squared and Wilcoxon rank-sum tests, respectively. Of the 29 patients who received mechanical support, 17 (58.6%) were discharged home, 11 (37.9%) died during their hospitalization, and 1 (3.5%) remains hospitalized. Median length of ventricular assist device support was 59.5 days (range 1-653 days). Between December 1991 and December 2011, in-hospital mortality was 64.3%. Following this period, significant changes were made to patient management with in-hospital mortality decreasing to 13.3% between February 2013 and December 2015. Comparison between deceased and living patients revealed several significant factors including: median number of packed red blood cells transfused, 8 versus 4 units (P = 0.048), median OR time, 396 versus 299 min (P = 0.003), and device implanted. During the early stages of the mechanical support program, higher than expected mortality rates prompted changes in the management of pediatric cardiac patients, specifically, the development of a dedicated management team. These changes significantly improved outcomes and we suggest can be used as a model for similar cardiac support programs, especially in smaller volume programs.


Toxicity of Cúspide 480SL® spray mixture formulation of glyphosate to aquatic organisms.
In 2011, an alternative formulation of glyphosate (Cúspide 480SL®) was chosen to replace Roundup-SL®, Fuete-SL®, and Gly-41® for the control of Erythroxylum coca, the source of cocaine, in Colombia. Cúspide 480SL contains the active ingredient glyphosate isopropylamine (IPA) salt, which is the same active ingredient used in previous formulations. However, Cúspide 480SL contains an alkyl polyglycoside surfactant rather than the polyethoxylated tallow amine (POEA) surfactant used in other formulations and known to be more toxic to nonprimary producing aquatic organisms than glyphosate itself. An adjuvant, Cosmo-Flux F411, and water also are added to the spray mixture before application. Aquatic ecosystems adjacent to the target coca fields might be exposed to the spray mix, placing aquatic organisms at risk. Because no toxicity data were available for spray mixture on aquatic organisms, acute toxicity tests were conducted on aquatic plants, invertebrates, and fish, by using the Cúspide 480SL spray mix as described on the label. Based on the median effective concentration (EC50) values for similar organisms, the spray mixture was less toxic to aquatic organisms than formulations previously used for the control of coca (i.e., Roundup-SL, Fuete-SL, and Gly-41). A physical effect induced by Cosmo-Flux F411 was observed in Daphnia magna, Ceriodaphnia dubia, and Hyalella azteca, causing the invertebrates to be trapped in an oily film that was present at the surface of the water. However, a hazard assessment for the Cúspide 480SL spray mix, using estimated worst-case exposure scenario concentrations and EC50 values from the toxicity tests, indicated de minimis hazard for the tested aquatic animals, with hazard quotients all <<1.


Extending the chemistry of weakly basic ligands: solvates of Ag+ and Cu+ stabilized by [Al{OC(CF3)3}4]- anion as model examples in the screening of useful weakly interacting solvents.
Weakly Coordinating Anions (WCAs) facilitate the formation of exotic "naked" cationic species. However, the feasibility of the respective synthesis approaches may be limited by the basicity of the solvent utilized, as the latter is one of the most important factors determining the solvation ability. In this work, we focus on a series of novel complexes of Ag(i) and Cu(i) with weakly basic ligands such as CH2Cl2, Cl3CCN and SO2 stabilized by perfluorinated alkoxyaluminate, Al[(ORF)4]-, RF = C(CF3)3. The discussion includes their synthesis protocols, crystal structures, vibrational spectra and thermal stability (TGA/DSC/EGA). We show that the Cu-SO2 adducts present exceptional stability in relation to other metal-SO2 complexes. To broaden the scope of weakly basic ligands which could prove helpful in the development of chemistry with WCAs, the screening of potential candidates based on DFT calculations is presented.


High-Fidelity Simulation Nurse Training Reduces Unplanned Interruption of Continuous Renal Replacement Therapy Sessions in Critically Ill Patients: The SimHeR Randomized Controlled Trial.
Although continuous renal replacement therapy (CRRT) is common, unplanned interruptions (UI) often limit its usefulness. In many units, nurses are responsible for CRRT management. We hypothesized that a nurse training program based on high-fidelity simulation would reduce the rate of interrupted sessions. We performed a 2-phase (training and evaluation), randomized, single-center, open study: During the training phase, intensive care unit nurses underwent a 6-hour training program and were randomized to receive (intervention) or not (control) an additional high-fidelity simulation training (6 hours). During the evaluation phase, management of CRRT sessions was randomized to either intervention or control nurses. Sessions were defined as UI if they were interrupted and the interruption was not prescribed in writing more than 3 hours before. Study nurses had experience with hemodialysis, but no experience with CRRT before training. Intervention nurses had higher scores than control nurses on the knowledge tests (grade, median [Q1-Q3], 14 [10.5-15] vs 11 [10-12]/20; P = .044). During a 13-month period, 106 sessions were randomized (n = 53/group) among 50 patients (mean age 70 ± 13 years, mean simplified acute physiology II score 69 [54-96]). Twenty-one sessions were not analyzed (4 were not performed and 17 patients died during sessions). Among the 42 intervention and 43 control sessions analyzed, 25 (59%) and 38 (88%) were labeled as UI (relative risk [95% CI], 0.67 [0.51-0.88]; P = .002). Intervention nurses required help significantly less frequently (0 [0-1] vs 3 [1-4] times/session; P < .0001). The 2 factors associated with UI in multilevel mixed-effects logistic regression were Sequential Organ Failure Assessment score (odds ratio [95% CI], 0.81 [0.65-99]; P = .047) and the intervention group (odds ratio, 0.19 [0.05-0.73]; P = .015). High-fidelity simulation nurse training reduced the rate of UI of CRRT sessions and the need for nurses to request assistance. This intervention may be particularly useful in the context of frequent nursing staff turnover.


Neutralizing antibodies to enterovirus 71 in Belém, Brazil.
Non-polio enteroviruses (Coxsackievirus A, Coxsackievirus B, Echovirus and EV 68-72) which belong to the enterovirus (EV) genus, Picornaviridae family, may be responsible for acute flaccid paralysis, aseptic meningitis, myocarditis, hepatitis, pleurodinia, neonatal sepsis, hand, foot and mouth disease (HFMD) even though 50-80% of infections are asymptomatic. EV 71 has been responsible for outbreaks and epidemics of HFMD and acute neurologic disease justifying its study in our country. The aim of this study was to detect neutralizing antibodies (NtAb) to EV 71 in individuals up to 15 years of age living in Belém, State of Pará, northern Brazil. Serum samples from 238 patients attending the Virology Sector of Evandro Chagas Institute in Belém, Brazil, were analyzed using microneutralization tests that included RD cells and BrCr strain. Overall 40.8% (97/238) of tested samples had NtAb to EV 71. Regarding the distribution per age group, 85.2% (92/108) of patients aged 0-3 years had no NtAb to this virus and 69.2% of those 12 to 15 years of age were seropositive. These results confirm that EV 71 infection occurs in the city of Belém; and that a high rate of individuals in this study were infected aged 3 years and over and, when aged 15 years nearly 70% had EV 71 NtAb.


Macrophages facilitate post myocardial infarction arrhythmias: roles of gap junction and KCa3.1.
Effective therapeutic targets against post-myocardial infarction (MI) arrhythmias remain to be discovered. We aimed to investigate the role of macrophages in post-MI arrhythmias. Methods: Mononuclear cell accumulation, macrophage polarization from M0 to M1 subset, and gap junction formation were analyzed in MI patients and MI mice by flow cytometry, immunofluorescence and patch clamping. Differentially expressed genes were identified by RNA sequencing. Macrophages and cardiomyocytes were cocultured in vitro, and the effects of gap junction and KCa3.1 on electrophysiological properties were assessed by patch clamping. The effects of KCa3.1 inhibition on post-MI arrhythmias were assessed by intracardiac stimulation and ambulatory electrocardiograms in vivo. Results: Percentage of pro-inflammatory mononuclear cells were significantly elevated in patients with post-MI arrhythmias compared with MI patients without arrhythmias and healthy controls (p<0.001). Macrophages formed gap junction with cardiomyocytes in MI border zones of MI patient and mice, and pro-inflammatory macrophages were significantly increased 3 days post-MI (p<0.001). RNA sequencing identified Kcnn4 as the most differentially expressed gene encoding ion channel, and the upregulation is mainly attributed to macrophage accumulation and polarization into pro-inflammatory subset. In vitro coculture experiments demonstrated that connection with M0 macrophages via gap junction slightly shortened the action potential durations (APDs) of cardiomyocytes. However, the APD90 of cardiomyocytes connected with M1 macrophages were significantly prolonged (p<0.001), which were effectively attenuated by gap junction inhibition (p=0.002), KCa3.1 inhibition (p=0.008), KCa3.1 silencing (p<0.001) and store-operated Ca2+ channel inhibition (p=0.005). In vivo results demonstrated that KCa3.1 inhibition significantly decreased the QTc durations (p=0.031), intracardiac stimulation-induced ventricular arrhythmia durations (p=0.050) and incidence of premature ventricular contractions (p=0.030) in MI mice. Conclusion: Macrophage polarization leads to APD heterogeneity and post-MI arrhythmias via gap junction and KCa3.1 activation. The results provide evidences of a novel mechanism of post-MI heterogeneous repolarization and arrhythmias, rendering macrophages and KCa3.1 to be potential therapeutic targets.


Human immunodeficiency virus replication induces monocyte chemotactic protein-1 in human macrophages and U937 promonocytic cells.
We have recently described a significant correlation between human immunodeficiency virus-1 (HIV-1) RNA replication and monocyte chemotactic protein-1 (MCP-1) levels in the cerebrospinal fluid (CSF) of individuals with the acquired immunodeficiency syndrome (AIDS) with HIV encephalitis (E). Because local macrophages (microglia) are the cells predominantly infected in the brain, we investigated whether in vitro HIV infection affects MCP-1 production in mononuclear phagocytes (MP). MCP-1 secretion and expression were consinstently upregulated over constitutive levels in human monocyte-derived macrophages (MDM) infected with the M-tropic R5 BaL strain of HIV-1. HIV replication was required for this effect, as demonstrated by the absence of chemokine upregulation after infection in the presence of 3'-azido-3'-deoxythimidine (AZT) or cell-exposure to heat-inactivated (triangle up degrees ) virus. MCP-1 induction was not restricted to HIV-1 BaL, but was also observed during productive infection of MDM with two primary isolates differing for entry coreceptor usage and of U937 cells with the X4 HIV-1 MN strain. Based on the observation that exogenous HIV-1 Tat induced MCP-1 expression in astrocytes, we also investigated its role in MDM and U937 cells. Exogenous Tat induced MCP-1 production from MDM in a concentration-dependent manner, however, it was not effective on uninfected U937 cells or on the chronically infected U937-derived cell line U1. Transfection of Tat-expressing plasmids moderately activated HIV expression in U1 cells, but failed to induce MCP-1 expression in this cell line or in uninfected U937 cells. HIV replication-dependent expression of MCP-1 in MP may be of particular relevance for the pathogenesis of HIV infection in nonlymphoid organs such as the brain.


Acoustophoretic focusing effects on particle synthesis and clogging in microreactors.
The handling of solids in microreactors represents a challenging task. In this paper, we present an acoustophoretic microreactor developed to manage particles in flow and to control the material synthesis process. The reactor was designed as a layered resonator with an actuation frequency of 1.21 MHz, in which a standing acoustic wave is generated in both the depth and width direction of the microchannel. The acoustophoretic force exerted by the standing wave on the particles focuses them to the channel center. A parametric study of the effect of flow rate, particle size and ultrasound conditions on the focusing efficiency was performed. Furthermore, the reactive precipitation of calcium carbonate and barium sulfate was chosen as a model system for material synthesis. The acoustophoretic focusing effect avoids solid deposition on the channel walls and thereby minimizes reactor fouling and thus prevents clogging. Both the average particle size and the span of the particle size distribution of the synthesized particles are reduced by applying high-frequency ultrasound. The developed reactor has the potential to control a wide range of material synthesis processes.


The Untapped Potential of the Nurse Practitioner Workforce in Reducing Health Disparities.
The growing nurse practitioner (NP) workforce represents a significant supply of primary care providers, who if optimally utilized, are well-positioned to improve access to health care for racial and ethnic minorities. However, many barriers affect the optimal utilization of NPs in primary care delivery. These barriers may also prevent NPs from maximally contributing to efforts to reduce racial and ethnic health disparities. Our review of the empirical and health policy literature sought to elucidate factors that affect NPs' potential and ability to narrow or eliminate health disparities. We found that restrictive state scope of practice regulations, disparate reimbursement policies, lack of NP workforce diversity, and poor organizational structures in NP practices may limit NPs' contributions to current efforts to reduce disparities. Our results led to the development of the nurse practitioner health disparities model which identifies barriers to and opportunities for optimal use of NPs in reducing racial and ethnic disparities. State and federal policymakers and administrators in health-care settings should take actions to remove legislative and organizational barriers to enable NPs to deliver high-quality care to racial and ethnic minorities. Researchers can use the nurse practitioner health disparities model to produce empirical evidence to reduce health disparities and improve population health.


Isolation and purification of gap junction channels.
This paper reports methods we have developed to solubilize gap junction channels, or connexons, from isolated gap junctions and to purify them in milligram quantities. Two sources of material are used: rat liver gap junctions and gap junctions produced by infecting insect cells with a baculovirus containing the cDNA for human liver beta 1 protein (connexin 32). Complete solubilization is obtained with long chain detergents (lauryl dimethyl amineoxide, dodecyl maltoside) and requires high ionic strength and high pH as well as reducing conditions. The purification involves chromatography on hydroxylapatite and gel filtration on Superose 6. A homogeneous product is indicated by a single band on a silver-stained gel and a homogeneous population of doughnut-shaped particles under the electron microscope. These particles have hexameric symmetry. The purified connexons have a tendency to form aggregates: filaments and sheets. The filaments grow by end-to-end association of connexons and are nonpolar, suggesting that the connexons are paired as in the cell-to-cell channel. The sheets grow by lateral association of the filaments.


Loss of muscle PDH induces lactic acidosis and adaptive anaplerotic compensation via pyruvate-alanine cycling and glutaminolysis.
Pyruvate dehydrogenase (PDH) is the rate-limiting enzyme for glucose oxidation that links glycolysis-derived pyruvate with the tricarboxylic acid (TCA) cycle. Although skeletal muscle is a significant site for glucose oxidation and is closely linked with metabolic flexibility, the importance of muscle PDH during rest and exercise has yet to be fully elucidated. Here, we demonstrate that mice with muscle-specific deletion of PDH exhibit rapid weight loss and suffer from severe lactic acidosis, ultimately leading to early mortality under low-fat diet provision. Furthermore, loss of muscle PDH induces adaptive anaplerotic compensation by increasing pyruvate-alanine cycling and glutaminolysis. Interestingly, high-fat diet supplementation effectively abolishes early mortality and rescues the overt metabolic phenotype induced by muscle PDH deficiency. Despite increased reliance on fatty acid oxidation during high-fat diet provision, loss of muscle PDH worsens exercise performance and induces lactic acidosis. These observations illustrate the importance of muscle PDH in maintaining metabolic flexibility and preventing the development of metabolic disorders.


A bright blue fluorescent dextran for two-photon in vivo imaging of blood vessels.
Fluorescence-based in vivo imaging is one of the most important tools for monitoring of biological processes in cells and tissues of live animal models. Fluorescence imaging agents have also been used to monitor the microcirculation. Tracking microcirculation of the blood is vital to gain further insight into various vascular disease-related anomalies within the human body. As monitoring of vascular circulation is performed with visualization of both immune cells and pathogens, which are mainly labelled with red and green, the favorable color option for blood vessels could be blue. However, currently available blueish color-labeled agents for vascular monitoring is generally confronted with quick bleaching, because of its short excitation and emission wavelengths. Hereby, what we propose in this report is a newly generated bright blue fluorescent dextran, named HCD-70K that monitors the blood vessels using blue and inter-compatible typical fluorescent materials. DBCO-functionalized dextran-70K was fabricated with hydroxy-coumarin dye via metal-free bioorthogonal click chemistry, and generated HCD-70K, which can flow within the blood vessel and decipher the whole structure of the blood vessel successfully. The synthesis, spectroscopic analysis, and quantum chemical calculations were conducted. Using two-photon microscopy, efficient deep in vivo blood vessel imaging of a mouse model revealed exceptional bio-imaging capabilities of the HCD-70K and consequently it provided a promising opportunity for efficient vascular visualization in various research areas.