ChatGPT identifies gender disparities in scientific peer review.
The peer review process is a critical step in ensuring the quality of scientific research. However, its subjectivity has raised concerns. To investigate this issue, I examined over 500 publicly available peer review reports from 200 published neuroscience papers in 2022-2023. OpenAI's generative artificial intelligence ChatGPT was used to analyze language use in these reports, which demonstrated superior performance compared to traditional lexicon- and rule-based language models. As expected, most reviews for these published papers were seen as favorable by ChatGPT (89.8% of reviews), and language use was mostly polite (99.8% of reviews). However, this analysis also demonstrated high levels of variability in how each reviewer scored the same paper, indicating the presence of subjectivity in the peer review process. The results further revealed that female first authors received less polite reviews than their male peers, indicating a gender bias in reviewing. In addition, published papers with a female senior author received more favorable reviews than papers with a male senior author, for which I discuss potential causes. Together, this study highlights the potential of generative artificial intelligence in performing natural language processing of specialized scientific texts. As a proof of concept, I show that ChatGPT can identify areas of concern in scientific peer review, underscoring the importance of transparent peer review in studying equitability in scientific publishing.


[Treatment of advanced hepatocellular carcinoma].
In the last few years, much progress has been made in the diagnosis and treatment of hepatocellular carcinoma (HCC). Due to these advances, HCC is no longer regarded as a disease with an extremely poor prognosis and has become the focus of some of the most active basic and clinical research in recent years. The most important advance is possibly the demonstration that sorafenib, a multikinase inhibitor with antiproliferative and antiangiogenic properties, is an effective treatment, able to increase survival in patients with advanced-stage HCC. This increased survival has demonstrated that these drugs, which act selectively on the molecular pathways involved in tumoral progression, can be effective in the treatment of HCC and has opened the door to the evaluation of these molecular agents, alone or in combination, in HCC. The present article provides a review of the treatment of advanced-stage HCC, with special emphasis on the distinct agents that are currently under evaluation.


Polymer-filled microcontainers for oral delivery loaded using supercritical impregnation.
In the last years a large variety of drug delivery systems have been developed to improve bioavailability of therapeutics in oral administration. An increasing interest has arisen in reservoir-based microdevices designed for active ingredients like water insoluble compounds and fragile biomolecules. Such microdevices are designed to protect the active ingredient against degradation and deactivation, and to allow cytoadhesion and unidirectional drug release. There are few works which optimize the drug loading step and often therapeutics are dosed in the microdevices through laborious and time consuming procedures. This work proposes an effective loading technique for a poorly soluble model drug in microcontainers, by combining inkjet printing and supercritical fluid impregnation. Well defined quantities of poly(vinyl pyrrolidone) (PVP) solutions are dispensed into microcontainers by inkjet printing with a quasi-no-waste performance. Then ketoprofen is impregnated in the polymer matrix by using supercritical carbon dioxide (scCO2) as loading medium. The amount of polymer is controlled by the volume and the number of droplets of dispensed polymer and drug loading is tuned by varying the impregnation parameters. Compared to solid dispersions of the same drug and polymer, scCO2-impregnated microcontainers exhibit a more reproducible drug loading and a faster dissolution rate of the active compound which allows drug release to be modulated. The combination of these loading techniques potentially allows the high throughput fabrication of microdevices for oral drug delivery with a safe and solvent-free solution.


Age and sex differences in the cerebellum and the ventral pons: a prospective MR study of healthy adults.
The human brain exhibits a complex pattern of differential aging. The purpose of this study was to examine whether age differences in the volume of cerebellar regions and the ventral pons are differential or generalized, whether the age-related shrinkage is linear or exponential, and whether there are sex differences in the size of the cerebellum and pons. The volumes of the cerebellar hemispheres (excluding the vermis and the peduncles), the vermis, and the ventral pons were estimated from the prospectively acquired MR scans of 190 healthy volunteers (aged 18-81 years). The relation between regional volumes, age, and sex was assessed while taking into account differences in body size (height). We found a moderate age-related reduction in the volume of the cerebellar hemispheres and the cerebellar vermis. In contrast to previous findings that suggested differential vulnerability of the posterior vermis, the age-related shrinkage of the vermian lobules was uniform-about 2% per decade. In accord with all reports in the literature, the size of the ventral pons was unrelated to age. The volume of the cerebellar hemispheres, the vermis, and the ventral pons were larger in men, even after adjustment for height. The magnitude of the sex difference was the largest in the hemispheres and the anterior vermis, and the smallest in the lobules VI-VII (declive-folium-tuber). Moderate age-related shrinkage of the cerebellum and lack of age-related differences in the ventral pons are robust phenomena. However, in all likelihood, the effects of age on the cerebellum are not differential but uniform. The cerebellum and the pons are larger in men than in women and the difference is especially pronounced in the cerebellar hemispheres and the anterior vermis.


Soluble Urokinase Receptor and Acute Kidney Injury.
Acute kidney injury is common, with a major effect on morbidity and health care utilization. Soluble urokinase plasminogen activator receptor (suPAR) is a signaling glycoprotein thought to be involved in the pathogenesis of kidney disease. We investigated whether a high level of suPAR predisposed patients to acute kidney injury in multiple clinical contexts, and we used experimental models to identify mechanisms by which suPAR acts and to assess it as a therapeutic target. We measured plasma levels of suPAR preprocedurally in patients who underwent coronary angiography and patients who underwent cardiac surgery and at the time of admission to the intensive care unit in critically ill patients. We assessed the risk of acute kidney injury at 7 days as the primary outcome and acute kidney injury or death at 90 days as a secondary outcome, according to quartile of suPAR level. In experimental studies, we used a monoclonal antibody to urokinase plasminogen activator receptor (uPAR) as a therapeutic strategy to attenuate acute kidney injury in transgenic mice receiving contrast material. We also assessed cellular bioenergetics and generation of reactive oxygen species in human kidney proximal tubular (HK-2) cells that were exposed to recombinant suPAR. The suPAR level was assessed in 3827 patients who were undergoing coronary angiography, 250 who were undergoing cardiac surgery, and 692 who were critically ill. Acute kidney injury developed in 318 patients (8%) who had undergone coronary angiography. The highest suPAR quartile (vs. the lowest) had an adjusted odds ratio of 2.66 (95% confidence interval [CI], 1.77 to 3.99) for acute kidney injury and 2.29 (95% CI, 1.71 to 3.06) for acute kidney injury or death at 90 days. Findings were similar in the surgical and critically ill cohorts. The suPAR-overexpressing mice that were given contrast material had greater functional and histologic evidence of acute kidney injury than wild-type mice. The suPAR-treated HK-2 cells showed heightened energetic demand and mitochondrial superoxide generation. Pretreatment with a uPAR monoclonal antibody attenuated kidney injury in suPAR-overexpressing mice and normalized bioenergetic changes in HK-2 cells. High suPAR levels were associated with acute kidney injury in various clinical and experimental contexts. (Funded by the National Institutes of Health and others.).


Uptake of histamine by mouse peritoneal macrophages and a macrophage cell line, RAW264.7.
We have previously demonstrated that dietary histamine is accumulated in the spleens of l-histidine decarboxylase (HDC)-deficient mice, which lack endogenous histamine synthesis. To characterize the clearance system for dietary histamine in mice, we investigated the cell type and mechanism responsible for histamine uptake in the spleens of HDC-deficient mice. Immunohistochemical analyses using an antihistamine antibody indicated that a portion of the CD14+ cells in the spleen is involved in histamine storage. Peritoneal macrophages obtained from Balb/c mice and a mouse macrophage cell line, RAW264.7, had potential for histamine uptake, which was characterized by a low affinity and high capacity for histamine. The histamine uptake by RAW264.7 cells was observed at physiological temperature and was potently inhibited by pyrilamine, chlorpromazine, quinidine, and chloroquine, moderately inhibited by N alpha-methylhistamine, dopamine, and serotonin, and not affected by tetraethylammonium and 1-methyl-4-phenylpyridinium. Intracellular histamine was not metabolized in RAW264.7 cells and was released at physiological temperature in the absence of extracellular histamine. These results suggest that histamine uptake by macrophages may be involved in the clearance of histamine in the local histamine-enriched environment.


A genome-wide association study identifies novel and functionally related susceptibility Loci for Kawasaki disease.
Kawasaki disease (KD) is a pediatric vasculitis that damages the coronary arteries in 25% of untreated and approximately 5% of treated children. Epidemiologic data suggest that KD is triggered by unidentified infection(s) in genetically susceptible children. To investigate genetic determinants of KD susceptibility, we performed a genome-wide association study (GWAS) in 119 Caucasian KD cases and 135 matched controls with stringent correction for possible admixture, followed by replication in an independent cohort and subsequent fine-mapping, for a total of 893 KD cases plus population and family controls. Significant associations of 40 SNPs and six haplotypes, identifying 31 genes, were replicated in an independent cohort of 583 predominantly Caucasian KD families, with NAALADL2 (rs17531088, p(combined) = 1.13 x 10(-6)) and ZFHX3 (rs7199343, p(combined) = 2.37 x 10(-6)) most significantly associated. Sixteen associated variants with a minor allele frequency of >0.05 that lay within or close to known genes were fine-mapped with HapMap tagging SNPs in 781 KD cases, including 590 from the discovery and replication stages. Original or tagging SNPs in eight of these genes replicated the original findings, with seven genes having further significant markers in adjacent regions. In four genes (ZFHX3, NAALADL2, PPP1R14C, and TCP1), the neighboring markers were more significantly associated than the originally associated variants. Investigation of functional relationships between the eight fine-mapped genes using Ingenuity Pathway Analysis identified a single functional network (p = 10(-13)) containing five fine-mapped genes-LNX1, CAMK2D, ZFHX3, CSMD1, and TCP1-with functional relationships potentially related to inflammation, apoptosis, and cardiovascular pathology. Pair-wise blood transcript levels were measured during acute and convalescent KD for all fine-mapped genes, revealing a consistent trend of significantly reduced transcript levels prior to treatment. This is one of the first GWAS in an infectious disease. We have identified novel, plausible, and functionally related variants associated with KD susceptibility that may also be relevant to other cardiovascular diseases.


Thawing boar semen in the presence of seminal plasma improves motility, modifies subpopulation patterns and reduces chromatin alterations.
Seminal plasma could have positive effects on boar semen after thawing. In the present study we investigated changes in the motility and chromatin structure in spermatozoa over 4h incubation (37°C) of boar semen thawed in the presence of 0%, 10% or 50% seminal plasma from good-fertility boars. Cryopreserved doses were used from seven males, three of which were identified as susceptible to post-thawing chromatin alterations. Motility was analysed by computer-aided sperm analysis every hour, and data were used in a two-step clustering, yielding three subpopulations of spermatozoa (slow non-linear, fast non-linear, fast linear). Chromatin structure was analysed using a sperm chromatin structure assay and flow cytometry to determine the DNA fragmentation index (%DFI) as a percentage, the standard deviation of the DFI (SD-DFI) and the percentage of high DNA stainability (%HDS), indicating chromatin compaction. Thawing without seminal plasma resulted in a rapid loss of motility, whereas seminal plasma helped maintain motility throughout the incubation period and preserved the subpopulation comprising fast and linear spermatozoa. The incidence of chromatin alterations was very low in samples from non-susceptible males, whereas samples from males susceptible to post-thawing chromatin alterations exhibited marked alterations in%DFI and%HDS. Seminal plasma partly prevented these alterations in samples from susceptible males. Overall, 50% seminal plasma was the most efficient concentration to protect motility and chromatin. Some changes were concomitant with physiological events reported previously (e.g., semen thawed with 50% seminal plasma increased the production of reactive oxygen species and yielded higher fertility after AI). Thawing in the presence of seminal plasma could be particularly useful in the case of samples susceptible to post-thawing chromatin damage.


The dynamics of protein body formation in developing wheat grain.
Wheat is a major source of protein in the diets of humans and livestock but we know little about the mechanisms that determine the patterns of protein synthesis in the developing endosperm. We have used a combination of enrichment with (15) N glutamine and NanoSIMS imaging to establish that the substrate required for protein synthesis is transported radially from its point of entrance in the endosperm cavity across the starchy endosperm tissues, before becoming concentrated in the cells immediately below the aleurone layer. This transport occurs continuously during grain development but may be slower in the later stages. Although older starchy endosperm cells tend to contain larger protein deposits formed by the fusion of small protein bodies, small highly enriched protein bodies may also be present in the same cells. This shows a continuous process of protein body initiation, in both older and younger starchy endosperm cells and in all regions of the tissue. Immunolabeling with specific antibodies shows that the patterns of enrichment are not related to the contents of gluten proteins in the protein bodies. In addition to providing new information on the dynamics of protein deposition, the study demonstrates the wider utility of NanoSIMS and isotope labelling for studying complex developmental processes in plant tissues.


Personality disorder risk factors for suicide attempts over 10 years of follow-up.
Identifying personality disorder (PD) risk factors for suicide attempts is an important consideration for research and clinical care alike. However, most prior research has focused on single PDs or categorical PD diagnoses without considering unique influences of different PDs or of severity (sum) of PD criteria on the risk for suicide-related outcomes. This has usually been done with cross-sectional or retrospective assessment methods. Rarely are dimensional models of PDs examined in longitudinal, naturalistic prospective designs. In addition, it is important to consider divergent risk factors in predicting the risk of ever making a suicide attempt versus the risk of making an increasing number of attempts within the same model. This study examined 431 participants who were followed for 10 years in the Collaborative Longitudinal Personality Disorders Study. Baseline assessments of personality disorder criteria were summed as dimensional counts of personality pathology and examined as predictors of suicide attempts reported at annual interviews throughout the 10-year follow-up period. We used univariate and multivariate zero-inflated Poisson regression models to simultaneously evaluate PD risk factors for ever attempting suicide and for increasing numbers of attempts among attempters. Consistent with prior research, borderline PD was uniquely associated with ever attempting. However, only narcissistic PD was uniquely associated with an increasing number of attempts. These findings highlight the relevance of both borderline and narcissistic personality pathology as unique contributors to suicide-related outcomes.


Multifunctional photosensitizer-based contrast agents for photoacoustic imaging.
Photoacoustic imaging is a novel hybrid imaging modality combining the high spatial resolution of optical imaging with the high penetration depth of ultrasound imaging. Here, for the first time, we evaluate the efficacy of various photosensitizers that are widely used as photodynamic therapeutic (PDT) agents as photoacoustic contrast agents. Photoacoustic imaging of photosensitizers exhibits advantages over fluorescence imaging, which is prone to photobleaching and autofluorescence interference. In this work, we examined the photoacoustic activity of 5 photosensitizers: zinc phthalocyanine, protoporphyrin IX, 2,4-bis [4-(N,N-dibenzylamino)-2,6-dihydroxyphenyl] squaraine, chlorin e6 and methylene blue in phantoms, among which zinc phthalocyanine showed the highest photoacoustic activity. Subsequently, we evaluated its tumor localization efficiency and biodistribution at multiple time points in a murine model using photoacoustic imaging. We observed that the probe localized at the tumor within 10 minutes post injection, reaching peak accumulation around 1 hour and was cleared within 24 hours, thus, demonstrating the potential of photosensitizers as photoacoustic imaging contrast agents in vivo. This means that the known advantages of photosensitizers such as preferential tumor uptake and PDT efficacy can be combined with photoacoustic imaging capabilities to achieve longitudinal monitoring of cancer progression and therapy in vivo.


The implementation of Health in All Policies initiatives: a systems framework for government action.
There has been a renewed interest in broadening the research agenda in health promotion to include action on the structural determinants of health, including a focus on the implementation of Health in All Policies (HiAP). Governments that use HiAP face the challenge of instituting governance structures and processes to facilitate policy coordination in an evidence-informed manner. Due to the complexity of government institutions and the policy process, systems theory has been proposed as a tool for evaluating the implementation of HiAP. Our multiple case study research programme (HiAP Analysis using Realist Methods On International Case Studies - HARMONICS) has relied on systems theory and realist methods to make sense of how and why the practices of policy-makers (including politicians and civil servants) from specific institutional environments (policy sectors) has either facilitated or hindered the implementation of HiAP. Herein, we present a systems framework for the implementation of HiAP based on our experience and empirical findings in studying this process. We describe a system of 14 components within three subsystems of government. Subsystems include the executive (heads of state and their appointed political elites), intersectoral (the milieu of policy-makers and experts working with governance structures related to HiAP) and intrasectoral (policy-makers within policy sectors). Here, HiAP implementation is a process involving interactions between subsystems and their components that leads to the emergence of implementation outcomes, as well as effects on the system components themselves. We also describe the influence of extra-governmental systems, including (but not limited to) the academic sector, third sector, private sector and intergovernmental sector. Finally, we present a case study that applies this framework to understand the implementation of HiAP - the Health 2015 Strategy - in Finland, from 2001 onward. This framework is useful for helping to explain how, why and under what circumstances HiAP has been successfully and unsuccessfully implemented in a sustainable manner. It serves as a tool for researchers to study this process, and for policy-makers and other public health actors to manage this process.


Acrylamide in home-prepared roasted potatoes.
Potato is one of the world's most widely grown tuber crop, in which starch is the predominant form of carbohydrates. Potatoes can be prepared in many ways: boiled, fried or roasted. Frying and roasting potatoes at high temperatures result in an appetizing crust, but at the same time acrylamide can form. In this study, the concentrations of the acrylamide precursors, asparagine and sugars, were determined in five different Swedish-grown potato varieties, together with the acrylamide content after typical home-cooking procedures; oven-roasting of potato wedges and pan-frying of cubes of boiled potatoes. Pan-frying of boiled potato cubes resulted in higher levels of acrylamide (530-1100 microg/kg) than in the wedges (140-250 microg/kg). Blanching combined with a shorter roasting time was shown to be an efficient way of reducing the acrylamide content in roasted potato wedges, especially in the experiments performed after long-term storage, where the acrylamide content was reduced from 110-260 to 50-140 microg/kg. No correlation was found between precursor content and acrylamide content, and this finding emphasizes the need for further studies on factors affecting acrylamide formation, for example, the availability of precursors at the surface during cooking.


The importance of a validated standard methodology to define in vitro toxicity of nano-TiO2.
Several in vitro studies on the potential toxicity of nano-TiO(2) have been published and recent reviews have summarised them. Most of these reports concluded that physicochemical properties of nanoparticles are fundamental to their toxicological effects. No published review has compared in vitro tests with similar test strategies in terms of exposure duration and measured endpoints and for this reason we have attempted to assess the degree of homogeneity among in vitro tests and to assess if they afford reliable data to support risk assessment. The responses in different in vitro tests appeared to be unrelated to primary particle size. The biologically effective concentrations in different tests can be seen to differ by as many as two orders of magnitude and such differences could be explained either by different sensitivities of cell lines to nanoparticles or by effect of the test media. Our review indicates that even when the in vitro tests measure the same biomarkers with the same exposure duration and known primary particle sizes, it is insufficient merely to use such data for risk assessment. In the future, validated standard methods should include a limited number of cell lines and an obligatory selection of biomarkers. For routine purposes, it is important that assays can be easily conducted, false negatives and false positives are excluded and unbiased interpretation of results is provided. Papers published to date provide an understanding of the mode on nano-TiO(2) action but are not suitable for assessment and management of risk.


Distribution of DNA damage in chromatin and its relation to repair in human cells treated with 7-bromomethylbenz(a) anthracene.
We have examined the relationship between the distribution of DNA damage and repair in chromatin from confluent human fibroblasts treated with the carcinogen 7-bromomethylbenz (a) anthracene. Analysis of staphylococcal nuclease (SN)4 digestion kinetics and gel electrophoresis revealed that more total damage occurs in nucleosome core DNA (approximately 80-85% of chromatin DNA) than in SN sensitive DNA (APPROXIMATELY15-20%). Furthermore, over a 24 hr period, damage is removed at about the same rate from these two regions. In contrast, virtually all of the nucleotides incorporated during repair synthesis are initially SN sensitive even when measured at 12 hr after damage. With time many repair-incorporated nucleotides become SN resistant and coelectrophorese with nucleosome core DNA. To explain these data we propose a model whereby excision repair occurs in both linker and core DNA; however, in core DNA the repair process induces conformational changes resulting in temporarily increased SN sensitivity; subsequently, rearrangement occurs and results in the re-establishment of native or near-native nucleosome conformation and SN resistance.


Repeated exposure of Caco-2 versus Caco-2/HT29-MTX intestinal cell models to (nano)silver in vitro: Comparison of two commercially available colloidal silver products.
Colloidal silver products are sold for a wide range of disinfectant and health applications. This has increased the potential for human exposure to silver nanoparticles (AgNPs) and ions (Ag+), for which oral ingestion is considered to be a major route of exposure. Our objective was to evaluate and compare the toxicity of two commercially available colloidal silver products on two human intestinal epithelial models under realistic exposure conditions. Mesosilver™ and AgC were characterized and a concentration range between 0.1 and 12 μg/mL chosen. Caco-2 cells vs. co-culture of Caco-2 and mucus-secreting HT29-MTX cells (90/10) were used. Repeated exposure was carried out to determine cell viability over 18 days of cell differentiation in 24-well plates. Selected concentrations (0.1, 1, and 3 μg/mL) were tested on cells cultured in E-plates and Transwells with the same repeated exposure regimen, to determine cell impedance, and cell viability and trans-epithelial electrical resistance (TEER), respectively. Silver uptake, intracellular localisation, and translocation were determined by CytoViva™, HIM-SIMS, and ICP-MS. Genotoxicity was determined on acutely-exposed proliferating Caco-2 cells by γH2AX immunofluorescence staining. Repeated exposure of a given concentration of AgC, which is composed solely of ionic silver, generally exerted more toxic effects on Caco-2 cells than Mesosilver™, which contains a mix of AgNPs and ionic silver. Due to its patchy structure, the presence of mucus in the Caco-2/HT29-MTX co-culture only slightly mitigated the deleterious effects on cell viability. Increased genotoxicity was observed for AgC on proliferating Caco-2 cells. Silver uptake, intracellular localisation, and translocation were similar. In conclusion, Mesosilver™ and AgC colloidal silver products show different levels of gut toxicity due to the forms of distinct silver (AgNPs and/or Ag+) contained within. This study highlights the applicability of high-resolution (chemical) imaging to detect and localize silver and provides insights into its uptake mechanisms, intracellular fate and cellular effects.


Conservative versus Interventional Treatment for Spontaneous Pneumothorax.
Whether conservative management is an acceptable alternative to interventional management for uncomplicated, moderate-to-large primary spontaneous pneumothorax is unknown. In this open-label, multicenter, noninferiority trial, we recruited patients 14 to 50 years of age with a first-known, unilateral, moderate-to-large primary spontaneous pneumothorax. Patients were randomly assigned to immediate interventional management of the pneumothorax (intervention group) or a conservative observational approach (conservative-management group) and were followed for 12 months. The primary outcome was lung reexpansion within 8 weeks. A total of 316 patients underwent randomization (154 patients to the intervention group and 162 to the conservative-management group). In the conservative-management group, 25 patients (15.4%) underwent interventions to manage the pneumothorax, for reasons prespecified in the protocol, and 137 (84.6%) did not undergo interventions. In a complete-case analysis in which data were not available for 23 patients in the intervention group and 37 in the conservative-management group, reexpansion within 8 weeks occurred in 129 of 131 patients (98.5%) with interventional management and in 118 of 125 (94.4%) with conservative management (risk difference, -4.1 percentage points; 95% confidence interval [CI], -8.6 to 0.5; P = 0.02 for noninferiority); the lower boundary of the 95% confidence interval was within the prespecified noninferiority margin of -9 percentage points. In a sensitivity analysis in which all missing data after 56 days were imputed as treatment failure (with reexpansion in 129 of 138 patients [93.5%] in the intervention group and in 118 of 143 [82.5%] in the conservative-management group), the risk difference of -11.0 percentage points (95% CI, -18.4 to -3.5) was outside the prespecified noninferiority margin. Conservative management resulted in a lower risk of serious adverse events or pneumothorax recurrence than interventional management. Although the primary outcome was not statistically robust to conservative assumptions about missing data, the trial provides modest evidence that conservative management of primary spontaneous pneumothorax was noninferior to interventional management, with a lower risk of serious adverse events. (Funded by the Emergency Medicine Foundation and others; PSP Australian New Zealand Clinical Trials Registry number, ACTRN12611000184976.).


The 'evolutionary signal' of homoplasy in protein-coding gene sequences and its consequences for a priori weighting in phylogeny.
To analyse independently homoplasy for the six possible types of substitution (i.e., A-G, C-T, A-C, A-T, C-G and G-T) at each of the three codon-positions of the cytochrome b gene, two approaches were used: the first is based on the consistency index which measures the amount of homoplasy, and the second is based on the saturation analysis which describes graphically the distribution of homoplasy within the taxonomic sampling. The results obtained from a data set of 32 sequences of Artiodactyla indicate that evolution of the cytochrome b is governed by differential constraints: 1) between the six substitutions-types, 2) between the three codon-positions, and 3) between the two mtDNA strands. Moreover, we find that non-synonymous sites can be more homoplastic than synonymous sites when the possibilities of substitutions are severely restricted because of the functional requirements of hydrophobicity. Most weighting schemes applied to protein-coding genes are elaborated from unjustified assumptions. We propose to weight each substitution-type at each codon-position according to its homoplasy content evaluated either with the consistency index or with an index representing the level of mutational saturation.


Chronic pancreatic inflammation induced by environmental tobacco smoke inhalation in rats.
Despite a strong epidemiological association between cigarette smoking and pancreatic diseases, such as pancreatic cancer and chronic pancreatitis, the effects of long-term cigarette smoke inhalation on the pancreas have not been clearly determined. In the present study, we investigated the effect of cigarette smoke inhalation on the pancreas. Thirty-six female Sprague Dawley rats were exposed to two different doses of environmental tobacco smoke averaging 100 mg or 160 mg/m3 total suspended particulate matter (TSP) per m3 for 70 min twice a day for 12 wk. The animals were sacrificed and examined for the effects of tobacco smoke exposure on pancreatic morphology and function. In 58% (7/12) of the animals, exposure to 160 mg/m3 TSP cigarette smoke induced a chronic pancreatic inflammatory process with fibrosis and scarring of pancreatic acinar structures. Animals with fibrotic alterations showed an induction of pancreatic pro-collagen 1 gene expression, and the infiltration of immune cells was accompanied by the expression of the inflammatory mediators MIP-1alpha, IL-1beta, and TGF-beta in 33% (4/12) of the animals. Acinar cell stress was manifested by a significant up-regulation of pancreatitis-associated protein expression (PAP) in smoke-exposed animals (smoke-exposed 6,932 +/- 1,236 vs control 3,608 +/- 305, p < 0.05). Possibly contributing to the morphological damage to the exocrine pancreas, the inhalation of cigarette smoke induced trypsinogen and chymotrypsinogen gene expression and, furthermore, reduced pancreatic enzyme content. This study provides experimental evidence of morphological pancreatic damage induced by the inhalation of cigarette smoke, which is likely to be mediated by alterations of acinar cell function.


Influence of stressors and possible pathways of onset of seventh graders' suicidal ideation in urban and rural areas in Taiwan.
Suicide is the second leading cause of death among young people in Taiwan. However, few studies have investigated children's suicidal ideation, and longitudinal studies are particularly rare. The purposes of this study were: (1) to describe the proportion of students with suicidal ideation in one month and incidence of suicidal ideation in the 7th graders (the first year of junior high school) living in urban and rural areas; (2) to realize the influence of perceived stressors on the onset of 7th graders' suicidal ideation; and (3) to explore possible pathways through which trigger factors and perceived stressors lead to suicidal ideation. A total of 1,589 students were followed from grade 4 to grade 7. Logistic regression was then used to investigate the influence of perceived stressors on the onset of suicidal ideation in grade 7. Structural equation modeling was used to analyze possible pathways through which trigger factors led to increased pressure from certain stressors which in turn resulted in suicidal ideation. The proportion of students with suicidal ideation in one month was 2 to 3 times higher in both areas compared to that in elementary school. However, the incidence in the rural area showed a large increase from 10.2% and 9.5% in grades 5 and 6 to 15.5% in grade 7. Urban-rural difference was observed. Important stressors and pathways of suicidal ideation differ between urban and rural areas. This study showed that the influential stressors in urban and rural areas might be different. Thus, interventions focused on coping skills for regional specific stressors and trigger factors could be beneficial in the transition time.


Hepatoprotective effect of thymol against subchronic toxicity of titanium dioxide nanoparticles: Biochemical and histological evidences.
The study was aimed to investigate the protective action of thymol against nano titanium dioxide (nano-TiO2) induced hepatotoxicity in rats. To achieve this purpose, the rats were divided into four groups (n = 6) including control, nano-TiO2 (100 mg/kg), nano-TiO2 + thymol (10 mg/kg) and nano-TiO2 + thymol (30 mg/kg). Intragastric (IG) administration of nano-TiO2 for 60 consecutive days caused widespread histological changes and significantly induced oxidative stress in the liver tissues as manifested by the rise in serum transaminase activities accompanied by marked decline of enzymatic (catalase, superoxide dismutase and glutathione peroxidase) and non-enzymatic (ferric reducing antioxidant power and glutathione) antioxidant levels, and rise of malondialdehyde levels in liver tissue. Pretreatment with thymol (IG) prior to nano-TiO2 administration significantly ameliorated all of biochemical and histopathological alterations in a dose-dependent manner. In conclusion, thymol effectively protects against nano-TiO2-induced hepatotoxicity in rats by its antioxidant properties.


Efficacy trial of an Internet-based intervention for cancer-related female sexual dysfunction.
The recent NCCN Guidelines for Survivorship recommend systematic evaluation and multidisciplinary treatment of cancer-related sexual dysfunctions. However, most oncology professionals fail to routinely assess sexual problems and lack expertise to treat them. An Internet-based intervention was designed to educate female patients and their partners about cancer-related sexual problems, describe medical treatment options and how to find expert care, and provide self-help strategies. A randomized trial assessed efficacy of the intervention when used as self-help versus the same Web access and 3 supplemental counseling sessions. Survivors of localized breast or gynecologic cancers completed online questionnaires at baseline, posttreatment, and 3- and 6-month follow-up, including the Female Sexual Function Index (FSFI), the Menopausal Sexual Interest Questionnaire (MSIQ), the Brief Symptom Inventory-18 (BSI-18) to assess emotional distress, and the Quality of Life in Adult Cancer Survivors (QLACS) scale. Program evaluation ratings were completed posttreatment. Fifty-eight women completed baseline questionnaires (mean age, 53 ± 9 years). Drop-out rates were 22% during treatment and 34% at 6-month follow-up. Linear mixed models for each outcome across time showed improvement in total scores on the FSFI, MSIQ, and QLACS (P<.001) and BSI-18 (P=.001). The counseled group improved significantly more on sexuality measures, but changes in emotional distress and quality of life did not differ between groups. Program content and ease of use were rated positively. Research is needed on how best to integrate this intervention into routine clinical practice, and particularly how to improve uptake and adherence.


Expression and functional analysis of Uch-L3 during mouse development.
Mice homozygous for the s(1Acrg) deletion at the Ednrb locus arrest at embryonic day 8.5. To determine the molecular basis of this defect, we initiated positional cloning of the s(1Acrg) minimal region. The mouse Uch-L3 (ubiquitin C-terminal hydrolase L3) gene was mapped within the s(1Acrg) minimal region. Because Uch-L3 transcripts were present in embryonic structures relevant to the s(1Acrg) phenotype, we created a targeted mutation in Uch-L3 to address its role during development and its possible contribution to the s(1Acrg) phenotype. Mice homozygous for the mutation Uch-L3(Delta3-7) were viable, with no obvious developmental or histological abnormalities. Although high levels of Uch-L3 RNA were detected in testes and thymus, Uch-L3(Delta3-7) homozygotes were fertile, and no defect in intrathymic T-cell differentiation was detected. We conclude that the s(1Acrg) phenotype is either complex and multigenic or due to the loss of another gene within the region. We propose that Uch-L3 may be functionally redundant with its homologue Uch-L1.


Effect of low vitamin A diets with high-moisture or dry corn on marbling and adipose tissue fatty acid composition of beef steers.
Angus-cross steers (n = 165; 295 +/- 16 kg of BW) were used evaluate the effect of low vitamin A diets with high-moisture corn (HMC) or dry corn (DC) on marbling and fatty acid composition. Steers were allotted to 24 pens (7 steers/pen), such that each pen had the same average initial BW. Treatments were randomly allotted to the pens. The experiment had a completely randomized design, with a 2 x 2 factorial arrangement of treatments: low vitamin A (Lo, no supplemental vitamin A) and HMC (LoHMC); LoDC; high vitamin A (Hi, supplemented with 2,200 IU of vitamin A/kg of DM) and HMC (HiHMC); and HiDC. Diets contained 76% corn, 10% corn silage, 11% protein supplement, and 3% soybean oil (DM basis). Samples of feed ingredients were collected for carotenoid analysis. Blood samples were collected for serum retinol determination. Steers were slaughtered after 145 d on feed. Carcass characteristics and LM composition were determined. Samples from the s.c. fat depot were analyzed for fatty acid composition. High-moisture corn had a greater vitamin A content, based on its carotenoid content, than DC (614 vs. 366 IU/kg of DM, P < 0.01). No vitamin A x corn type interactions were detected for feedlot performance, carcass characteristics, or serum, s.c. fat, or liver retinol concentration. Average daily gain, DMI, and G:F were not affected by vitamin A (P > 0.05). Marbling score and USDA quality grade were greater (P < 0.05) in Lo vs. Hi steers. Hot carcass weight, backfat, and yield grade were not affected by the treatments (P > 0.05). Vitamin A and corn type did not affect LM composition (DM, ash, CP, or ether-extractable fat, P > 0.05). Vitamin A supplementation increased (P < 0.06) serum retinol on d 112 and 145 and increased (P < 0.01) liver retinol at slaughter (Lo = 38.7 vs. Hi = 102.9 mug/g). The s.c. fat retinol concentrations were less (P < 0.01) for Lo (0.8 mug/g) than for Hi (1.4 mug/g) at slaughter. Cell diameter of adipocytes in the i.m. depot was not affected by dietary vitamin A (P > 0.05). A vitamin A x corn type interaction was observed (P < 0.05) for the s.c. fat cellularity. Feeding HMC increased the number of cells per square millimeter when Lo diets were fed (LoHMC = 128 vs. LoDC = 100 cells/mm(2), P < 0.05), but not when Hi diets were fed (HiHMC = 109 vs. HiDC = 111 cells/mm(2), P > 0.05). The CLA content of adipose tissue was not affected by the treatments. Regardless of the corn type used, feeding low vitamin A diets for 145 d to Angus-cross steers increased marbling and quality grade without affecting yield grade, animal health, or performance.


Whole Cell Cross-Linking to Discover Host-Microbe Protein Cognate Receptor/Ligand Pairs.
Bacterial surface ligands mediate interactions with the host cell during association that determines the specific outcome for the host-microbe association. The association begins with receptors on the host cell binding ligands on the microbial cell to form a partnership that initiates responses in both cells. Methods to determine the specific cognate partnerships are lacking. Determining these molecular interactions between the host and microbial surfaces are difficult, yet crucial in defining biologically important events that are triggered during association of the microbiome, and critical in defining the initiating signal from the host membrane that results in pathology or commensal association. In this study, we designed an approach to discover cognate host-microbe receptor/ligand pairs using a covalent cross-linking strategy with whole cells. Protein/protein cross-linking occurred when the interacting molecules were within 9-12 Å, allowing for identification of specific pairs of proteins from the host and microbe that define the molecular interaction during association. To validate the method three different bacteria with three previously known protein/protein partnerships were examined. The exact interactions were confirmed and led to discovery of additional partnerships that were not recognized as cognate partners, but were previously reported to be involved in bacterial interactions. Additionally, three unknown receptor/ligand partners were discovered and validated with in vitro infection assays by blocking the putative host receptor and deleting the bacterial ligand. Subsequently, Salmonella enterica sv. Typhimurium was cross-linked to differentiated colonic epithelial cells (caco-2) to discover four previously unknown host receptors bound to three previously undefined host ligands for Salmonella. This approach resulted in a priori discovery of previously unknown and biologically important molecules for host/microbe association that were casually reported to mediate bacterial invasion. The whole cell cross-linking approach promises to enable discovery of possible targets to modulate interaction of the microbiome with the host that are important in infection and commensalism, both of with initiate a host response.


Attaching and effacing (A/E) lesion formation by enteropathogenic E. coli on human intestinal mucosa is dependent on non-LEE effectors.
Enteropathogenic E. coli (EPEC) is a human pathogen that causes acute and chronic pediatric diarrhea. The hallmark of EPEC infection is the formation of attaching and effacing (A/E) lesions in the intestinal epithelium. Formation of A/E lesions is mediated by genes located on the pathogenicity island locus of enterocyte effacement (LEE), which encode the adhesin intimin, a type III secretion system (T3SS) and six effectors, including the essential translocated intimin receptor (Tir). Seventeen additional effectors are encoded by genes located outside the LEE, in insertion elements and prophages. Here, using a stepwise approach, we generated an EPEC mutant lacking the entire effector genes (EPEC0) and intermediate mutants. We show that EPEC0 contains a functional T3SS. An EPEC mutant expressing intimin but lacking all the LEE effectors but Tir (EPEC1) was able to trigger robust actin polymerization in HeLa cells and mucin-producing intestinal LS174T cells. However, EPEC1 was unable to form A/E lesions on human intestinal in vitro organ cultures (IVOC). Screening the intermediate mutants for genes involved in A/E lesion formation on IVOC revealed that strains lacking non-LEE effector/s have a marginal ability to form A/E lesions. Furthermore, we found that Efa1/LifA proteins are important for A/E lesion formation efficiency in EPEC strains lacking multiple effectors. Taken together, these results demonstrate the intricate relationships between T3SS effectors and the essential role non-LEE effectors play in A/E lesion formation on mucosal surfaces.


Factors impacting the acceptance of traceability in the food supply chain in the United States of America.
Global demands for increased food safety and quality assurance programmes, increased global competition, changing government rules and regulations, political and trade barriers, bioterrorism, and identity preservation requirements in global markets are affecting the world's food supply chain. To satisfy changing market demands, all suppliers in the food supply chain must adaptto these global issues. Total asset visibility must be maintained in production, in process, in storage, and in transit.


Complete reconstitution of the human coenzyme A biosynthetic pathway via comparative genomics.
The biosynthesis of CoA from pantothenic acid (vitamin B5) is an essential universal pathway in prokaryotes and eukaryotes. The CoA biosynthetic genes in bacteria have all recently been identified, but their counterparts in humans and other eukaryotes remained mostly unknown. Using comparative genomics, we have identified human genes encoding the last four enzymatic steps in CoA biosynthesis: phosphopantothenoylcysteine synthetase (EC ), phosphopantothenoylcysteine decarboxylase (EC ), phosphopantetheine adenylyltransferase (EC ), and dephospho-CoA kinase (EC ). Biological functions of these human genes were verified using a complementation system in Escherichia coli based on transposon mutagenesis. The individual human enzymes were overexpressed in E. coli and purified, and the corresponding activities were experimentally verified. In addition, the entire pathway from phosphopantothenate to CoA was successfully reconstituted in vitro using a mixture of purified recombinant enzymes. Human recombinant bifunctional phosphopantetheine adenylyltransferase/dephospho-CoA kinase was kinetically characterized. This enzyme was previously suggested as a point of CoA biosynthesis regulation, and we have observed significant differences in mRNA levels of the corresponding human gene in normal and tumor cells by Northern blot analysis.


Violent crime, suicide, and premature mortality in patients with schizophrenia and related disorders: a 38-year total population study in Sweden.
People with schizophrenia and related disorders are at an increased risk of adverse outcomes, including conviction of a violent offence, suicide, and premature mortality. However, the rates of, and risk factors for, these outcomes need clarification as a basis for population-based and targeted interventions. We aimed to determine rates and risk factors for these outcomes, and investigate to what extent they are shared across outcomes and are specific to schizophrenia and related disorders. We undertook a total population cohort study in Sweden of 24 297 patients with schizophrenia and related disorders between January, 1972 and December, 2009. Patients were matched by age and sex to people from the general population (n=485 940) and also to unaffected sibling controls (n=26 357). First, we investigated rates of conviction of a violent offence, suicide, and premature mortality, with follow-up until conviction of a violent offence, emigration, death, or end of follow-up (Dec 31, 2009), whichever occurred first. Second, we analysed associations between these adverse outcomes and sociodemographic, individual, familial, and distal risk factors, for men and women separately, with Cox proportional hazards models. Finally, we assessed time trends in adverse outcomes between 1972 and 2009, for which we compared patients with unaffected siblings, and analysed associations with changes in the number of nights spent in inpatient beds in psychiatric facilities nationwide. Within 5 years of their initial diagnosis, 13·9% of men and 4·7% of women with schizophrenia and related disorders had a major adverse outcome (10·7% of men and 2·7% of women were convicted of a violent offence, and 3·3% of men and 2·0% of women died prematurely of any cause). During the study, the adjusted odds ratio of any adverse outcomes for patients compared with general population controls was 7·5 (95% CI 7·2-7·9) in men and 11·1 (10·2-12·1) in women. Three risk factors that were present before diagnosis were predictive of any adverse outcome: drug use disorders, criminality, and self-harm, which were also risk factors for these outcomes in unaffected siblings and in the general population. Over the period 1973-2009, the odds of these outcomes increased in patients with schizophrenia and related disorders compared with unaffected siblings. Schizophrenia and related disorders are associated with substantially increased rates of violent crime, suicide, and premature mortality. Risk factors for these three outcomes included both those specific to individuals with schizophrenia and related disorders, and those shared with the general population. Therefore, a combination of population-based and targeted strategies might be necessary to reduce the substantial rates of adverse outcomes in patients with schizophrenia and related disorders.


A probabilistic approach to the interpretation of milk antibody results for diagnosis of Johne's disease in dairy cattle.
Johne's disease is a serious wasting disease of ruminants that is of high economic importance for the dairy sector in particular. The chronic nature of the disease, the fluctuations in antibody levels and the limited ability of diagnostic tests to identify cows at early stages of infection are huge challenges for the control of the disease. In the United Kingdom, the latter is commonly based on repeated milk ELISA testing of lactating cows, followed by selected culling and improved management practices around calving. In this paper, the dataset built through a large quarterly screening programme conducted in the United Kingdom since 2010 is used to investigate the use of milk ELISA testing for Johne's disease management. Over the study period, 13,509 out of 281,558 cows were identified as high-risk of being infected and shedding mycobacteria in the faeces, based on a case definition of at least two consecutive positive milk ELISA results. Around a third of them were kept in the dairy herd a year or more after being classified as high-risk. However, 16% of these cows did not have any further positive test, suggesting that they might be uninfected animals. The mean specificity and sensitivity of the milk ELISA test were estimated at 99.5% and 61.8%, respectively. The cows in the dataset are categorised in different result groups according to the number of positive test results and whether they are classified as high-risk according to the programme's case definition. The posterior probability of infection is calculated after each test in order to investigate the impact of repeated testing on the belief in a cow's infection status. The interpretation of the results show that most cows classified as high-risk are very likely to be infected, while some other groups that do not match the case definition could reasonably be considered as infected too. Our results show that there is considerable potential for more targeted use of serological testing, including adjusting the testing frequency and implementing the posterior probability approach.


The use of ultrasensitive quantitative-PCR to assess the impact of primaquine on asymptomatic relapse of Plasmodium vivax infections: a randomized, controlled trial in Lao PDR.
Trials to assess the efficacy of the radical cure of Plasmodium vivax malaria with 8-aminoquinolines require that most post-treatment relapses are identified, but there is no consensus on the optimal duration of follow-up in either symptomatic or asymptomatic vivax malaria. The efficacy of a 14-day course of primaquine on the cumulative incidence of recurrent asymptomatic P. vivax infections detected by ultrasensitive quantitative PCR (uPCR) as a primary endpoint was assessed. A randomized, placebo-controlled, single-blind trial was conducted in four villages of the Lao PDR during 2016-2018 nested in a larger project evaluating mass drug administrations (MDA) with dihydroartemisinin-piperaquine (DP) and a single low-dose primaquine to clear Plasmodium falciparum infections. In the nested sub-study, eligible participants with mono- or mixed P. vivax infections detected by uPCR were randomized to receive either 14 days of primaquine (0.5 mg/kg/day) or placebo during the last round of MDA (round 3) through directly observed therapy. Participants were checked monthly for 12 months for parasitaemia using uPCR. The primary outcome was cumulative incidence of participants with at least one recurrent episode of P. vivax infection. 20 G6PD-normal participants were randomized in each arm. 5 (29%) of 20 participants in the placebo arm experienced asymptomatic, recurrent P. vivax infections, resulting in a cumulative incidence at month 12 of 29%. None of the 20 participants in the intervention arm had recurrent infections (p = 0.047 Fisher's exact test). Participants with recurrent P. vivax infections were found to be parasitaemic for between one and five sequential monthly tests. The median time to recurrence of P. vivax parasitaemia was 178 days (range 62-243 days). A 14-day course of primaquine in addition to a DP-MDA was safe, well-tolerated, and prevented recurrent asymptomatic P. vivax infections. Long follow-up for up to 12 months is required to capture all recurrences following the treatment of asymptomatic vivax infection. To eliminate all malarias in settings where P. vivax is endemic, a full-course of an 8-aminoquinolines should be added to MDA to eliminate all malarias. Trial registration This study was registered with ClinicalTrials.gov under NCT02802813 on 16th June 2016. https://clinicaltrials.gov/ct2/show/NCT02802813.


The experiential impact of hospitalisation: Parents' accounts of caring for young people with early psychosis.
This research examines the experiential impact of hospitalisation on the parents of young people with early psychosis. In-depth interviews were conducted with a small sample of parents, and the resulting transcripts were analysed using Interpretative Phenomenological Analysis. Five themes emerged from the data: Accepting and blaming, Feeling out of control, Hospitalisation as temporary containment, Feeling let down by services and Stigma. Aspects of the hospitalisation process were characterised by parents as generally negative, but a number of positive affirmations were also offered regarding the containing, supportive and crucial role of services. Parents' perceptions of hospitalisation as a difficult, and sometimes distressing, experience are exacerbated by the complexity of being the carer of a young person. Negotiating services and boundaries within the context of this relationship contributes to feelings of exclusion and disregard by professionals and services. The implications of this study resonate with the current government mental health strategy with regard to how services can engage and include carers in the mental health system, and equip and enable them to support their relatives with early psychosis.


Recommendations from a Portuguese Expert Group for Discontinuation of Tyrosine Kinase Inhibitors in Chronic Myeloid Leukemia Patients in Clinical Practice.
Until recently, the main goal of chronic myeloid leukemia therapy was disease control with the best overall survival, which required lifelong treatment. However, currently, the treatment-free remission concept is becoming an important goal in clinical practice, and several tyrosine kinase inhibitors discontinuation studies have shown that round 50% of patients with a durable deep molecular response beyond major molecular response successfully interrupt tyrosine kinase inhibitors for at least three years without loss of molecular response. However, and regardless of the existing evidence, the exact conditions for attempting treatment-free remission remain poorly defined. Different authors tried to guide the clinical decision regarding this topic but there are some points that differ, namely with respect to the recommended duration of tyrosine kinase inhibitors therapy and the appropriate molecular response prior to treatment-free remission. The goal of this article is to propose an algorithm to guide clinical practice in Portugal concerning chronic phase-chronic myeloid leukemia patients who wish to attempt treatment-free remission, since there are no national guidelines.


A meta-analysis of studies on attitudes toward bears and wolves across Europe 1976-2012.
The ranges of wolves (Canis lupus) and bears (Ursus arctos) across Europe have expanded recently, and it is important to assess public attitudes toward this expansion because responses toward these species vary widely. General attitudes toward an object are good predictors of broad behavioral patterns; thus, attitudes toward wolves and bears can be used as indicators to assess the social foundation for future conservation efforts. However, most attitude surveys toward bears and wolves are limited in scope, both temporally and spatially, and provide only a snapshot of attitudes. To extend the results of individual surveys over a much larger temporal and geographical range so as to identify transnational patterns and changes in attitudes toward bears and wolves over time, we conducted a meta-analysis. Our analysis included 105 quantitative surveys conducted in 24 countries from 1976 to 2012. Across Europe, people's attitudes were more positive toward bears than wolves. Attitudes toward bears became more positive over time, but attitudes toward wolves seemed to become less favorable the longer people coexisted with them. Younger and more educated people had more positive attitudes toward wolves and bears than people who had experienced damage from these species, and farmers and hunters had less positive attitudes toward wolves than the general public. For bears attitudes among social groups did not differ. To inform conservation of large carnivores, we recommend that standardized longitudinal surveys be established to monitor changes in attitudes over time relative to carnivore population development. Our results emphasize the need for interdisciplinary research in this field and more advanced explanatory models capable of capturing individual and societal responses to changes in large carnivore policy and management.


Self-reported fever and measured temperature in emergency department records used for syndromic surveillance.
Many public health agencies monitor population health using syndromic surveillance, generally employing information from emergency department (ED) visit records. When combined with other information, objective evidence of fever may enhance the accuracy with which surveillance systems detect syndromes of interest, such as influenza-like illness. This study found that patient chief complaint of self-reported fever was more readily available in ED records than measured temperature and that the majority of patients with an elevated temperature recorded also self-reported fever. Due to its currently limited availability, we conclude that measured temperature is likely to add little value to self-reported fever in syndromic surveillance for febrile illness using ED records.


Atrial peptide natriuresis in the rat without genuine rise in filtration rate or wash-out of medullary electrolytes.
1. Effects of synthetic atrial natriuretic peptide (ANP) on renal excretion, total renal blood flow (RBF), glomerular filtration rate (GFR) and tissue electrical admittance (reciprocal impedance, an estimate of tissue electrolytes) were determined in pentobarbitone-anaesthetized rats. GFR was measured both as inulin clearance (Cin) and as a product of renal plasma flow (RPF) and inulin extraction ratio (Ein). 2. With the lowest dose of ANP (0.35 micrograms/(kg min) I.V.) a 5-fold increase in sodium excretion occurred without measurable change in Cin, RPF x Ein nor medullary electrolyte concentration estimated from tissue electrical admittance. 3. With medium and high dosage (2 and 6 micrograms/(kg min), respectively), major and rapid increases in sodium excretion and urine flow were associated with an acute increase in Cin but not RPF x Ein. 4. The RBF increase observed in all groups of rats was not dose-related and did not parallel the natriuresis. Electrolyte concentration in the medullary tissue showed a modest transient decrease in rats given medium and high ANP doses. 5. We conclude that pronounced ANP natriuresis can develop in the absence of a measurable increase of GFR, estimated by a method not subject to urinary dead space error (RPF x Ein). The small transient decrease in medullary tissue electrolytes observed with higher peptide doses does not support solute wash-out as an important mechanism of increased sodium excretion.


The Fate of Sulfamethazine in Sodium-Hypochlorite-Treated Drinking Water: Monitoring by LC-MS (n) -IT-TOF.
Pharmaceutical compounds represent a rapidly emerging class of environmental contaminants. Such compounds were recently classified by the U.S. Geological Survey, including several antibiotics. An LC-MS/MS screening method for the top five antibiotics in drinking water was developed and validated using a Shimadzu LC-MS-IT-TOF. The separation was performed using a Waters Acquity UPLC BEH C18 column with a gradient elution. Sulfamethazine was exposed to conditions intended to mimic drinking water chlorination, and samples were collected and quenched with excess sodium sulfite. Kinetics of sulfamethazine degradation was followed as well as the formation of the major chlorinated byproduct (m/z 313). For the screening method, all five antibiotic peaks were baseline resolved within 5 minutes. Additionally, precision and accuracy of the screening method were less than 15%. Degradation of sulfamethazine upon exposure to drinking water chlorination occurred by first order kinetics with a half-life of 5.3 × 10(4) min (approximately 37 days) with measurements starting 5 minutes after chlorination. Likewise, the formation of the major chlorinated product occurred by first order kinetics with a rate constant of 2.0 × 10(-2). The proposed identification of the chlorinated product was 4-amino-(5-chloro-4,6-dimethyl-2-pyrimidinyl)-benzenesulfonamide (C12H13N4O2SCl) using MS (n) spectra and databases searches of SciFinder and ChemSpider.


Duplications and losses of genes encoding known elements of the stress defence system of the Aspergilli contribute to the evolution of these filamentous fungi but do not directly influence their environmental stress tolerance.
The contribution of stress protein duplication and deletion events to the evolution of the Aspergilli was studied. We performed a large-scale homology analysis of stress proteins and generated and analysed three stress defence system models based on Saccharomyces cerevisiae, Schizosaccharomyces pombe and Aspergillus nidulans. Although both yeast-based and A. nidulans-based models were suitable to trace evolutionary changes, the A. nidulans-based model performed better in mapping stress protein radiations. The strong Mantel correlation found between the positions of species in the phylogenetic tree on the one hand and either in the A. nidulans-based or S. cerevisiae-based models on the other hand demonstrated that stress protein expansions and reductions contributed significantly to the evolution of the Aspergilli. Interestingly, stress tolerance attributes correlated well with the number of orthologs only for a few stress proteins. Notable examples are Ftr1 iron permease and Fet3 ferro-O2-oxidoreductase, elements of the reductive iron assimilation pathway, in the S. cerevisiae-based model, as well as MpkC, a HogA-like mitogen activated protein kinase in the A. nidulans-based model. In the case of the iron assimilation proteins, the number of orthologs showed a positive correlation with H2O2-induced stress tolerance while the number of MpkC orthologs correlated positively with Congo Red induced cell wall stress, sorbitol induced osmotic stress and H2O2 induced oxidative stress tolerances. For most stress proteins, changes in the number of orthologs did not correlate well with any stress tolerance attributes. As a consequence, stress tolerance patterns of the studied Aspergilli did not correlate with either the sets of stress response proteins in general or with the phylogeny of the species studied. These observations suggest that stress protein duplication and deletion events significantly contributed to the evolution of stress tolerance attributes of Aspergilli. In contrast, there are other processes, which may counterbalance the effects of stress gene duplications or deletions including (i) alterations in the structures of stress proteins leading to changes in their biological activities, (ii) varying biosynthesis of stress proteins, (iii) rewiring stress response regulatory networks or even (iv) acquiring new stress response genes by horizontal gene transfer. All these multilevel changes are indispensable for the successful adaptation of filamentous fungi to altering environmental conditions, especially when these organisms are entering new ecological niches.


Gastrin activates autophagy and increases migration and survival of gastric adenocarcinoma cells.
The peptide hormone gastrin exerts a growth-promoting effect in both normal and malignant gastrointestinal tissue. Gastrin mediates its effect via the cholecystokinin 2 receptor (CCKBR/CCK2R). Although a substantial part of the gastric adenocarcinomas express gastrin and CCKBR, the role of gastrin in tumor development is not completely understood. Autophagy has been implicated in mechanisms governing cytoprotection, tumor growth, and contributes to chemoresistance. This study explores the role of autophagy in response to gastrin in gastric adenocarcinoma cell lines. Immunoblotting, survival assays and the xCELLigence system were used to study gastrin induced autophagy. Chemical inhibitors of autophagy were utilized to assess the role of this process in the regulation of cellular responses induced by gastrin. Further, knockdown studies using siRNA and immunoblotting were performed to explore the signaling pathways that activate autophagy in response to gastrin treatment. We demonstrate that gastrin increases the expression of the autophagy markers MAP1LC3B-II and SQSTM1 in gastric adenocarcinoma cells. Gastrin induces autophagy via activation of the STK11-PRKAA2-ULK1 and that this signaling pathway is involved in increased migration and cell survival. Furthermore, gastrin mediated increase in survival of cells treated with cisplatin is partially dependent on induced autophagy. This study reveals a novel role of gastrin in the regulation of autophagy. It also opens up new avenues in the treatment of gastric cancer by targeting CCKBR mediated signaling and/or autophagy in combination with conventional cytostatic drugs.


Electron and hole transfer dynamics of a triarylamine-based dye with peripheral hole acceptors on TiO2 in the absence and presence of solvent.
We investigated photoinduced primary charge transfer processes of the sensitizer E6 on TiO2 without solvent and in contact with the organic solvent acetonitrile and the ionic liquid 1-ethyl-3-methylimidazolium tetracyanoborate [C2mim](+)[B(CN)4](-) using transient absorption spectroscopy, spectroelectrochemistry, and DFT/TDDFT calculations. E6, which belongs to a family of triarylamine dyes for solar cell applications, features two peripheral triarylamine units which are connected via diether spacer groups to the core chromophore and are designed to act as hole traps. This function was confirmed by spectroelectrochemistry, where the E6˙(+) radical cation shows a considerably blue-shifted absorption compared to dyes without these two substituents. This indicates that one of the terminal triarylamine units must carry the positive charge. After photoexcitation of E6 at 520 nm (S0 → S1 band), electrons are injected into TiO2 predominantly within the cross-correlation time (<80 fs), with some subsequent delayed electron injection (τ ca. 250 fs). Importantly, a transient Stark shift (electrochromism) is observed (time constants ca. 0.8 and 12 ps) which is related to a changing electric field generated by the E6˙(+) radical cations and injected electrons. This field induces absorption shifts of the dye species on the surface. Interestingly, these dynamics are largely unaffected by solvent molecules. However, pronounced differences are observed on longer timescales. In contact with solvent, one observes an increase in the E6˙(+) absorption band above 600 nm with a time constant of 75 ps. This is assigned to hole transfer from the core chromophore to one of the peripheral triarylamine substituents. Electron-cation recombination occurs on much longer timescales and is multiexponential, with time constants of ca. 100 μs, 1 ms and 15 ms. Because of hole trapping, it is slower than for similar dyes lacking the peripheral triarylamines. Additional experiments were performed for E6 attached to the wide band gap semiconductor ZrO2. Here, electron injection occurs into surface trap states with subsequent recombination. Another fraction of non-injecting E6 molecules in S1 quickly decays to S0 (time constants 1 and 35 ps).


Profiling of Glucose-Sensing Neurons Reveals that GHRH Neurons Are Activated by Hypoglycemia.
Comprehensive transcriptional profiling of glucose-sensing neurons is challenging because of low expression levels of glucokinase (Gck) and other key proteins that transduce a glucose signal. To overcome this, we generated and validated transgenic mice with a neuronal/endocrine-specific Gck promoter driving cre expression and mated them to mice with cre-dependent expression of an EGFP-tagged ribosomal protein construct (EEF1A1-LSL.EGFPL10) that can be used to map and profile cells. We found significant Gck expression in hypothalamic and limbic regions in cells that are activated following administration of glucose or 2-deoxyglucose. Transcriptional profiling from Gck-cre/EEF1A1-LSL.EGFPL10 mice enriched known and previously unknown glucose-sensing populations including neurons expressing growth hormone releasing hormone (GHRH). Electrophysiological recordings show that hypoglycemia activates GHRH neurons, suggesting a mechanistic link between hypoglycemia and growth hormone release. These studies provide a means for mapping glucose-sensitive neurons and for generating transcriptional profiles from other cell types expressing cre in a cell-specific manner.


Urinary bladder mass in a dog.
A 7-year-old spayed female Schnauzer dog with chronic hematuria had a soft tissue mass within the wall of the bladder. The mass was excised and, when examined histologically, was determined to be a discrete well-organized mural mass consisting of spindloid cells arranged in swirling sheets and palisades punctuated by aggregates of principally eosinophils. The overlying transitional mucosa was extensively ulcerated and focally hyperplastic and nodular with subjacent solid down growths, superficial cysts, and the extension of tubular structures deep into the submucosa. The histologic appearance of this mass is consistent with canine polypoid eosinophilic cystitis, also known as benign inflammatory fibrous polyp.


A novel membrane based process to isolate photosystem-I membrane complex from spinach.
The isolation of photosystem-I (PS-I) from spinach has been conducted using ultrafiltration with 300 kDa molecular weight cut-off polyethersulfone membranes. The effects of ultrafiltration operating conditions on PS-I activity were optimized using parameter scanning ultrafiltration. These conditions included solution pH, ionic strength, stirring speed, and permeate flux. The effects of detergent (Triton X-100 and n-dodecyl-beta-D-maltoside) concentration on time dependent activity of PS-I were also studied using an O(2) electrode. Under optimized conditions, the PS-I purity obtained in the retentate was about 84% and the activity recovery was greater than 94% after ultrafiltration. To our knowledge, this is the first report of the isolation of a membrane protein using ultrafiltration alone.


Systematic structural characterization of metabolites in Arabidopsis via candidate substrate-product pair networks.
Plant metabolomics is increasingly used for pathway discovery and to elucidate gene function. However, the main bottleneck is the identification of the detected compounds. This is more pronounced for secondary metabolites as many of their pathways are still underexplored. Here, an algorithm is presented in which liquid chromatography-mass spectrometry profiles are searched for pairs of peaks that have mass and retention time differences corresponding with those of substrates and products from well-known enzymatic reactions. Concatenating the latter peak pairs, called candidate substrate-product pairs (CSPP), into a network displays tentative (bio)synthetic routes. Starting from known peaks, propagating the network along these routes allows the characterization of adjacent peaks leading to their structure prediction. As a proof-of-principle, this high-throughput cheminformatics procedure was applied to the Arabidopsis thaliana leaf metabolome where it allowed the characterization of the structures of 60% of the profiled compounds. Moreover, based on searches in the Chemical Abstract Service database, the algorithm led to the characterization of 61 compounds that had never been described in plants before. The CSPP-based annotation was confirmed by independent MS(n) experiments. In addition to being high throughput, this method allows the annotation of low-abundance compounds that are otherwise not amenable to isolation and purification. This method will greatly advance the value of metabolomics in systems biology.


Advancing Community-Based Falls Prevention Programs for Older Adults-The Work of the Administration for Community Living/Administration on Aging.
The mission of the Administration for Community Living (ACL) is to maximize the independence, well-being, and health of older adults, people with disabilities across the lifespan, and their families and caregivers. In direct alignment with this mission is ACL's support of evidence-based falls prevention programs in communities throughout the United States. Since 2014, the Administration on Aging (AoA), part of ACL, has invested nearly $14 million in entities such as state agencies, nonprofits, and universities to expand access to proven community-based falls prevention programs. The initiatives supported by ACL/AoA bring to bear two primary goals-(1) to significantly increase the number of older adults and older adults with disabilities at risk for falls who participate in evidence-based community programs to reduce falls and falls risks; and (2) to implement innovative funding arrangements, including contracts, partnerships, and collaborations with one or more sustainability partners to support these programs during and beyond the grant period. Support from ACL/AoA has significantly increased the availability of evidence-based falls prevention programs in funded communities, as well as enhanced the network's sustainable delivery infrastructure to promote continued access to these critical programs beyond the scope of grant funding. This article highlights the successful rollout of ACL/AoA's falls prevention initiative.


Application of hyperbaric oxygen in liver transplantation.
In recent years, hyperbaric oxygen (HBO) has been used in the treatment of a lot of diseases such as decompression sickness, arterial gas embolism, carbon dioxide poisoning, soft tissue infection, refractory osteomyelitis, and problematic wound, but little is known about its application in liver transplantation. Although several studies have been conducted to investigate the protective effects of HBO on liver transplantation and liver preservation, there are still some controversies on this issue, especially its immunomodulatory effect. In this short review, we briefly summarize the findings supporting the application of HBO during liver transplantation (including donors and recipients).


The RNA-Recognition Motifs of TAR DNA-Binding Protein 43 May Play a Role in the Aberrant Self-Assembly of the Protein.
The TAR DNA-binding protein 43 (TDP-43) is a nucleic acid-binding protein implicated in gene regulation and RNA processing and shuffling. It is a ribonuclear protein that carries out most of its functions by binding specific nucleic acid sequences with its two RNA-recognition motifs, RRM1 and RRM2. TDP-43 has been identified in toxic cytosolic inclusions in neurodegenerative diseases such as amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration with ubiquitin-positive inclusions (FTLD-U). The unstructured C-terminus has prion-like behavior and has been considered the driver of the aberrant self-assembly of TDP-43. In this work, we set out to test the hypothesis that the RNA-binding domains could also play a role in protein aggregation. This knowledge could be of important value for understanding TDP-43 aberrant, disease-leading behavior and, in the future, inform the design of small molecules that could prevent or slow down protein aggregation by exploiting the RNA-binding properties of the protein. We investigated the behavior of the two tandem RRM domains separately and linked together and studied their self-assembly properties and RNA-binding ability with a number of biophysical techniques. The picture that emerges from our study suggests that this region of the protein plays an important and so far unexplored role in the aggregation of this protein.


A high-performance nanoporous Si/Al2O3 foam lithium-ion battery anode fabricated by selective chemical etching of the Al-Si alloy and subsequent thermal oxidation.
Nanostructured micrometer-sized Al-Si particles are synthesized via a facile selective etching process of Al-Si alloy powder. Subsequent thin Al2O3 layers are introduced on the Si foam surface via a selective thermal wet oxidation process of etched Al-Si particles. The resulting Si/Al2O3 foam anodes exhibit outstanding cycling stability (a capacity retention of 78% after 300 cycles at the C/5 rate) and excellent rate capability.


Outcomes after ecallantide treatment of laryngeal hereditary angioedema attacks.
Hereditary angioedema (HAE) is a rare disorder associated with episodic attacks of well-demarcated angioedema. Attacks that affect the larynx can result in life-threatening airway obstruction. To examine efficacy and safety of ecallantide treatment for laryngeal HAE attacks. Data were combined from 4 clinical studies (EDEMA2, EDEMA3, EDEMA4, and DX-88/19) evaluating 30 mg of subcutaneous ecallantide for treatment of acute HAE attacks. Efficacy was assessed using 2 validated, HAE-specific, patient-reported outcome measures. The change in Mean Symptom Complex Severity (MSCS) score indicates change in symptom severity; a negative score indicates improvement. The calculated minimally important difference (MID) for change in severity is -0.30. The Treatment Outcome Score (TOS) measures treatment response. A positive score indicates improvement; the calculated MID is 30. Overall, 98 patients received ecallantide for 220 laryngeal attacks. The mean ± SD change in MSCS score was -1.1 ± 0.73 and -1.6 ± 0.68 at 4 and 24 hours, respectively. The mean ± SD TOS was 73.5 ± 35.8 and 85.5 ± 27.8 at 4 and 24 hours, respectively. Median time to significant improvement was 185 minutes (95% confidence interval, 167-226). One attack required intubation. Four treatment-emergent serious adverse events were reported, including 2 HAE attacks that resulted in hospitalization and 2 anaphylactic reactions. One of these reactions required treatment with epinephrine, but both patients recovered fully. There were no deaths. In this large attack series, ecallantide was effective for treatment of laryngeal HAE attacks. There is a risk of hypersensitivity, including anaphylaxis, consistent with product labeling. As such, ecallantide should be administered under the supervision of a health care professional. clinicaltrials.gov Identifiers: not applicable for EDEMA2 (trial was conducted before implementation of registration requirements); NCT00262080 for EDEMA3, NCT00457015 for EDEMA4, and NCT00456508 for DX-88/19.


Origin of the linearity no threshold (LNT) dose-response concept.
This paper identifies the origin of the linearity at low-dose concept [i.e., linear no threshold (LNT)] for ionizing radiation-induced mutation. After the discovery of X-ray-induced mutations, Olson and Lewis (Nature 121(3052):673-674, 1928) proposed that cosmic/terrestrial radiation-induced mutations provide the principal mechanism for the induction of heritable traits, providing the driving force for evolution. For this concept to be general, a LNT dose relationship was assumed, with genetic damage proportional to the energy absorbed. Subsequent studies suggested a linear dose response for ionizing radiation-induced mutations (Hanson and Heys in Am Nat 63(686):201-213, 1929; Oliver in Science 71:44-46, 1930), supporting the evolutionary hypothesis. Based on an evaluation of spontaneous and ionizing radiation-induced mutation with Drosophila, Muller argued that background radiation had a negligible impact on spontaneous mutation, discrediting the ionizing radiation-based evolutionary hypothesis. Nonetheless, an expanded set of mutation dose-response observations provided a basis for collaboration between theoretical physicists (Max Delbruck and Gunter Zimmer) and the radiation geneticist Nicolai Timoféeff-Ressovsky. They developed interrelated physical science-based genetics perspectives including a biophysical model of the gene, a radiation-induced gene mutation target theory and the single-hit hypothesis of radiation-induced mutation, which, when integrated, provided the theoretical mechanism and mathematical basis for the LNT model. The LNT concept became accepted by radiation geneticists and recommended by national/international advisory committees for risk assessment of ionizing radiation-induced mutational damage/cancer from the mid-1950s to the present. The LNT concept was later generalized to chemical carcinogen risk assessment and used by public health and regulatory agencies worldwide.


Effects of Zoospore Concentration and Application Pressure on Foliage Blight of Catharanthus roseus Caused by Phytophthora nicotianae.
This study investigated the effects of inoculum concentration, application pressure, use of carbon dioxide (CO2), and agitation associated with passage through a spray nozzle on zoospore survival and on foliage blight caused by Phytophthora nicotianae. In a greenhouse experiment, plants of Catharanthus roseus were inoculated by spraying zoospore suspensions at pressures of 210, 420, and 630 kPa (30, 60, and 90 lbs/in2 [psi]). A low-pressure 35-kPa (5-psi) control inoculation was provided with a hand-operated aspirator-type mister. There was a consistent reduction in level of disease with increased application pressure, regardless of the zoospore concentration. To determine the source of this disease reduction, laboratory assays were conducted. Zoospore suspensions were pressurized to 210, 420, and 630 kPa with CO2 or air, then transferred from the pressure bottle into a flask by either spraying or pouring. From the flask, the suspensions were spread over plates of PARP-V8 agar and incubated for 72 h, at which time total colony numbers were recorded. CO2 significantly reduced zoospore survival. Pressure strength and method of spore transfer out of pressure bottles also impacted survival to lesser extents. There were significant interactions between pressure source and means of spore suspension transfer, and between pressure strength and means of spore transfer. These results may lead to development of alternative methods of water decontamination to prevent inoculum from entering crop systems through irrigation water.


Micellization behavior of coarse grained surfactant models.
We use molecular dynamics simulations over microsecond time scales to study the micellization behavior of recently proposed continuum-space, coarse grained surfactant models. In particular, we focus on the MARTINI model by Marrink et al. [J. Phys. Chem. B 111, 7812 (2007)] and a model by Shinoda et al. [Soft Matter 4, 2454 (2008)]. We obtain the critical micelle concentration (cmc) and equilibrium aggregate size distributions at low surfactant loadings. We present evidence justifying modest extrapolations for determining the cmc at low temperatures, where significant sampling difficulties remain. The replica exchange method provides only modest improvements of sampling efficiency for these systems. We find that the two coarse grained models significantly underpredict experimental cmc near room temperature for zwitterionic surfactants, but are closer to measured values for nonionic ones. The aggregation numbers for both zwitterionic and nonionic surfactants are near those observed experimentally, but the temperature dependence of the cmc is incorrect in both cases, because of the use of an unstructured solvent. Possible refinements to the models to bring them into quantitative agreement with experiment are discussed.


Reduced impact of pyrimethamine drug pressure on Plasmodium malariae dihydrofolate reductase gene.
Molecular investigations performed following the emergence of sulfadoxine-pyrimethamine (SP) resistance in Plasmodium falciparum have allowed the identification of the dihydrofolate reductase (DHFR) enzyme as the target of pyrimethamine. Although clinical cases of Plasmodium malariae are not usually treated with antifolate therapy, incorrect diagnosis and the high frequency of undetected mixed infections has probably exposed non-P. falciparum parasites to antifolate therapy in many areas. In this context, we aimed to assess the worldwide genetic diversity of the P. malariae dhfr gene in 123 samples collected in Africa and Asia, areas with different histories of SP use. Among the 10 polymorphic sites found, we have observed 7 new mutations (K55E, S58R, S59A, F168S, N194S, D207G, and T221A), which led us to describe 6 new DHFR proteins. All isolates from African countries were classified as wild type, while new mutations and haplotypes were recognized as exclusive to Madagascar (except for the double mutations at nucleotides 341 and 342 [S114N] found in one Cambodian isolate). Among these nonsynonymous mutations, two were likely related to pyrimethamine resistance: S58R (corresponding to C59R in P. falciparum and S58R in Plasmodium vivax; observed in one Malagasy sample) and S114N (corresponding to S108N in P. falciparum and S117N in P. vivax; observed in three Cambodian samples).


Quality improvement collaborative aiming for Proactive HEAlthcare of Older People in Care Homes (PEACH): a realist evaluation protocol.
This protocol describes a study of a quality improvement collaborative (QIC) to support implementation and delivery of comprehensive geriatric assessment (CGA) in UK care homes. The QIC will be formed of health and social care professionals working in and with care homes and will be supported by clinical, quality improvement and research specialists. QIC participants will receive quality improvement training using the Model for Improvement. An appreciative approach to working with care homes will be encouraged through facilitated shared learning events, quality improvement coaching and assistance with project evaluation. The QIC will be delivered across a range of partnering organisations which plan, deliver and evaluate health services for care home residents in four local areas of one geographical region. A realist evaluation framework will be used to develop a programme theory informing how QICs are thought to work, for whom and in what ways when used to implement and deliver CGA in care homes. Data collection will involve participant observations of the QIC over 18 months, and interviews/focus groups with QIC participants to iteratively define, refine, test or refute the programme theory. Two researchers will analyse field notes, and interview/focus group transcripts, coding data using inductive and deductive analysis. The key findings and linked programme theory will be summarised as context-mechanism-outcome configurations describing what needs to be in place to use QICs to implement service improvements in care homes. The study protocol was reviewed by the National Health Service Health Research Authority (London Bromley research ethics committee reference: 205840) and the University of Nottingham (reference: LT07092016) ethics committees. Both determined that the Proactive HEAlthcare of Older People in Care Homes study was a service and quality improvement initiative. Findings will be shared nationally and internationally through conference presentations, publication in peer-reviewed journals, a graphical illustration and a dissemination video.


Staffing With Disease-Based Epidemiologic Indices May Reduce Shortage of Intensive Care Unit Staff During the COVID-19 Pandemic.
Health care worker (HCW) safety is of pivotal importance during a pandemic such as coronavirus disease 2019 (COVID-19), and employee health and well-being ensure functionality of health care institutions. This is particularly true for an intensive care unit (ICU), where highly specialized staff cannot be readily replaced. In the light of lacking evidence for optimal staffing models in a pandemic, we hypothesized that staff shortage can be reduced when staff scheduling takes the epidemiology of a disease into account. Various staffing models were constructed, and comprehensive statistical modeling was performed. A typical routine staffing model was defined that assumed full-time employment (40 h/wk) in a 40-bed ICU with a 2:1 patient-to-staff ratio. A pandemic model assumed that staff worked 12-hour shifts for 7 days every other week. Potential in-hospital staff infections were simulated for a total period of 120 days, with a probability of 10%, 25%, and 40% being infected per week when at work. Simulations included the probability of infection at work for a given week, of fatality after infection, and the quarantine time, if infected. Pandemic-adjusted staffing significantly reduced workforce shortage, and the effect progressively increased as the probability of infection increased. Maximum effects were observed at week 4 for each infection probability with a 17%, 32%, and 38% staffing reduction for an infection probability of 0.10, 0.25, and 0.40, respectively. Staffing along epidemiologic considerations may reduce HCW shortage by leveling the nadir of affected workforce. Although this requires considerable efforts and commitment of staff, it may be essential in an effort to best maintain staff health and operational functionality of health care facilities and systems.


A fatal drug interaction between oxycodone and clonazepam.
A case is presented of a fatal drug interaction caused by ingestion of oxycodone (Oxycontin) and clonazepam (Klonapin). Oxycodone is an opium alkaloid used in long-term pain management therapy. Clonazepam is a benzodiazepine used for the treatment of seizures and panic disorders. The Drug Abuse Warning Network (DAWN) has reported an increase of 108% in the last two years of emergency department episodes related to Oxycontin. Six billion prescriptions were written for Oxycontin in the year 2000, an 18-fold increase from four years previous (1). Oxycontin has recently gained enormous notoriety at the local and national levels; however, there are very few previously documented cases of lethal drug interactions between oxycodone and clonazepam. Synergistic effects between these two drugs are postulated to arise from different agonistic mechanisms producing similar physiological changes. It is also theorized that clonazepam may inhibit the metabolism of oxycodone. A 38-year-old white female was found dead in Jefferson County, Tennessee in March of 2001. The deceased had physical evidence of previous drug abuse and positive serological findings of hepatitis B and C. Prescription pill bottles filled under the name of the deceased, as well as another name, were found with the body. Serum, urine and gastric contents from the deceased were screened for numerous drugs and metabolites using a combination of thin layer chromatography and immunoassay techniques (EMIT and FPIA). Analysis of biological specimens from the deceased revealed the presence of: benzodiazepines, opiates (oxycodone), and trazodone metabolites in the serum; cannabinoids, benzodiazepines, opiates (oxycodone), trazodone, trazodone metabolites, nicotine, and nicotine metabolite in the urine; and benzodiazepines, opiates (oxycodone), nicotine, and nicotine metabolite in the gastric contents. Quantitative analyses for clonazepam was performed by high performance liquid chromatography (HPLC) and revealed a plasma concentration of 1.41 microg/mL. Plasma oxycodone and urine 11-nor-carboxy-delta-9-tetrahydrocannabinol concentrations were determined by gas chromatography/mass spectrometry and revealed concentrations of 0.60 microg/mL and 27.9 ng/mL, respectively. The deceased had pathologies consistent with severe central nervous system (CNS) and respiratory depression produced by high concentrations of clonazepam and oxycodone including collapsed lungs, aspirated mucus, and heart failure. The pathologies were sufficient to cause death, which was officially attributed to a drug overdose; however, the manner of death was unknown.


Study of the mechanism of vanadate inhibition of the dynein cross-bridge cycle in sea urchin sperm flagella.
The effect of vanadate on the ATP-induced disruption of trypsin-treated axonemes and the ATP-induced straightening of rigor wave preparations of sea urchin sperm was investigated. Addition of ATP to a suspension of trypsin-treated axonemes results in a rapid decrease in turbidity (optical density measured at 350 nm) concomitant with the disruption of the axonemes by sliding between microtubules to form tangles of connected doublet microtubules (Summers and Gibbons, 1971; Sale and Satir, 1977). For axonemes digested to approximately 93 percent of their initial turbidity, 5 {muM} vanadate completely inhibits the ATP-induced decrease in turbidity and the axonemes maintain their structural integrity. However, with axonemes digested to approximately 80 percent of their initial turbidity, vanadate fails to inhibit the ATP-induced decrease in turbidity and the ATP-induced structural disruption of axonemes, even when the vanadate concentration is raised as high as 100 mum. For such axonemes digested to 80 percent of their initial turbidity, the form of ATP-induced structural changes, in the presence of 25 muM vanadate, was observed by dark-field light microscopy and revealed that the axonemes become disrupted into curved, isolated doublet microtubules, small groups of doublet microtubules, and "banana peel" structures in which tubules have peeled back from the axoneme. Addition of 5 muM ATP to rigor wave sperm, which were prepared by abrupt removal of ATP from reactivated sperm, causes straightening of the rigor waves within 1 min, and addition of more than 10 muM ATP causes resumption of flagellar beating. Addition of 40 muM vanadate to the rigor wave sperm does not inhibit straightening of the rigor waves of 2 muM-1 mM ATP, although oscillatory beating is completely inhibited. These results suggest that vanadate inhibits the mechanochemical cycle of dyein at a step subsequent to the MgATP(2-)-induced release of the bridged dynein arms.