CEPS: An Open Access MATLAB Graphical User Interface (GUI) for the Analysis of Complexity and Entropy in Physiological Signals.
We developed CEPS as an open access MATLAB® GUI (graphical user interface) for the analysis of Complexity and Entropy in Physiological Signals (CEPS), and demonstrate its use with an example data set that shows the effects of paced breathing (PB) on variability of heart, pulse and respiration rates. CEPS is also sufficiently adaptable to be used for other time series physiological data such as EEG (electroencephalography), postural sway or temperature measurements. Data were collected from a convenience sample of nine healthy adults in a pilot for a larger study investigating the effects on vagal tone of breathing paced at various different rates, part of a development programme for a home training stress reduction system. The current version of CEPS focuses on those complexity and entropy measures that appear most frequently in the literature, together with some recently introduced entropy measures which may have advantages over those that are more established. Ten methods of estimating data complexity are currently included, and some 28 entropy measures. The GUI also includes a section for data pre-processing and standard ancillary methods to enable parameter estimation of embedding dimension m and time delay τ ('tau') where required. The software is freely available under version 3 of the GNU Lesser General Public License (LGPLv3) for non-commercial users. CEPS can be downloaded from Bitbucket. In our illustration on PB, most complexity and entropy measures decreased significantly in response to breathing at 7 breaths per minute, differentiating more clearly than conventional linear, time- and frequency-domain measures between breathing states. In contrast, Higuchi fractal dimension increased during paced breathing. We have developed CEPS software as a physiological data visualiser able to integrate state of the art techniques. The interface is designed for clinical research and has a structure designed for integrating new tools. The aim is to strengthen collaboration between clinicians and the biomedical community, as demonstrated here by using CEPS to analyse various physiological responses to paced breathing.


Calcium homeostasis in human melanocytes: role of transient receptor potential melastatin 1 (TRPM1) and its regulation by ultraviolet light.
Transient receptor potential melastatin (TRPM) is a subfamily of ion channels that are involved in sensing taste, ambient temperature, low pH, osmolarity, and chemical ligands. Melastatin 1/TRPM1, the founding member, was originally identified as melanoma metastasis suppressor based on its expression in normal pigment cells in the skin and the eye but not in aggressive, metastasis-competent melanomas. The role of TRPM1 and its regulation in normal melanocytes and in melanoma progression is not understood. Here, we studied the relationship of TRPM1 expression to growth and differentiation of human epidermal melanocytes. TRPM1 expression and intracellular Ca(2+) levels are significantly lower in rapidly proliferating melanocytes compared to the slow growing, differentiated melanocytes. We show that lentiviral short hairpin RNA (shRNA)-mediated knockdown of TRPM1 results in reduced intracellular Ca(2+) and decreased Ca(2+) uptake suggesting a role for TRPM1 in Ca(2+) homeostasis in melanocytes. TRPM1 knockdown also resulted in a decrease in tyrosinase activity and intracellular melanin pigment. Expression of the tumor suppressor p53 by transfection or induction of endogenous p53 by ultraviolet B radiation caused repression of TRPM1 expression accompanied by decrease in mobilization of intracellular Ca(2+) and uptake of extracellular Ca(2+). These data suggest a role for TRPM1-mediated Ca(2+) homeostasis, which is also regulated by ultraviolet B, in melanogenesis.


An integrative review of information and communication technology based support interventions for carers of home dwelling older people.
A growing number of studies of informal carers of older people reveal positive results concerning support via Information and Communication Technologies (ICT). Systematic examination of factors that have a potential impact on carer outcomes are needed to inform future research. To explore studies concerning ICT support of adult carers of older people and to identify study characteristics that have a potential impact on carer outcomes. This integrative review includes 123 studies published since 2005. Fundamental questions for designing sensitive support interventions; 'who, what, and how' were applied to a synthesis of the results. Identified characteristics from the studies responding to the who question included variables of the carers, such as their relationship with the care recipient or their ethnicity. Characteristics related to the what question related to the types of interventions, and the how question concerned the different services or programs offered, the idiosyncratic needs of the carers, and the types of technologies used. Results are discussed according to micro, meso and macro levels of analysis. This extensive review can inform future studies and highlight the evidence in the area for decision makers, practitioners and/or NGOs working with innovative forms of support for carers of older people.


[Knowledge, opinions and cigarette smoking among teachers].
Due to the importance that educational activities have in preventing the habit of smoking in boys and young men, a research was carried out in order to know the rates of predominance, knowledge and opinions teachers have on smoking. For this purpose, an enquiry was applied to 746 teachers from Osorno, Chile. The results showed a rate of 42.6% of smokers; considering specialization, the major predominance is found in woman teachers of Biology and Physical Education. Comparisons with predominance in other countries are established. It was determined that the level of knowledge is independent from the habit of smoking; on the contrary, opinions about different aspects related to smoking, are closely connected to the habit. We conclude that it is necessary to carry on a systematic training with in service teachers and specially in Teacher's Training Colleges.


Large-scale organization of ferret auditory cortex revealed using continuous acquisition of intrinsic optical signals.
We have adapted a new approach for intrinsic optical imaging, in which images were acquired continuously while stimuli were delivered in a series of continually repeated sequences, to provide the first demonstration of the large-scale tonotopic organization of both primary and nonprimary areas of the ferret auditory cortex. Optical responses were collected during continuous stimulation by repeated sequences of sounds with varying frequency. The optical signal was averaged as a function of time during the sequence, to produce reflectance modulation functions (RMFs). We examined the stability and properties of the RMFs and show that their zero-crossing points provide the best temporal reference points for quantifying the relationship between the stimulus parameter values and optical responses. Sequences of different duration and direction of frequency change gave rise to comparable results, although in some cases discrepancies were observed, mostly between upward- and downward-frequency sequences. We demonstrated frequency maps, consistent with previous data, in primary auditory cortex and in the anterior auditory field, which were verified with electrophysiological recordings. In addition to these tonotopic gradients, we demonstrated at least 2 new acoustically responsive areas on the anterior and posterior ectosylvian gyri, which have not previously been described. Although responsive to pure tones, these areas exhibit less tonotopic order than the primary fields.


Illustrative context-preserving exploration of volume data.
In volume rendering, it is very difficult to simultaneously visualize interior and exterior structures while preserving clear shape cues. Highly transparent transfer functions produce cluttered images with many overlapping structures, while clipping techniques completely remove possibly important context information. In this paper, we present a new model for volume rendering, inspired by from illustration. It provides a means of interactively inspecting the interior of a volumetric data set in a feature-driven way which retains context information. The context-preserving volume rendering model uses a function of shading intensity, gradient magnitude, distance to the eye point, and previously accumulated opacity to selectively reduce the opacity in less important data regions. It is controlled by two user-specified parameters. This new method represents an alternative to conventional clipping techniques, sharing their easy and intuitive user control, but does not suffer from the drawback of missing context information.


Atrial arrhythmia, triggering events and conduction abnormalities in isolated murine RyR2-P2328S hearts.
RyR2 mutations are associated with catecholaminergic polymorphic tachycardia, a condition characterized by ventricular and atrial arrhythmias. The present experiments investigate the atrial electrophysiology of homozygotic murine RyR2-P2328S (RyR2(S/S)) hearts for ectopic triggering events and for conduction abnormalities that might provide a re-entrant substrate. Electrocardiograph recordings were made from regularly stimulated RyR2(S/S) and wild type (WT) hearts, perfused using a novel modified Langendorff preparation. This permitted the simultaneous use of either floating intracellular microelectrodes to measure action potential (AP) parameters, or a multielectrode array to measure epicardial conduction velocity (CV). RyR2(S/S) showed frequent sustained tachyarrhythmias, delayed afterdepolarizations and ectopic APs, increased interatrial conduction delays, reduced epicardial CVs and reduced maximum rates of AP depolarization ((dV/dt)(max)), despite similar effective refractory periods, AP durations and AP amplitudes. Effective interatrial CVs and (dV/dt)(max) values of APs following ectopic (S2) stimulation were lower than those of APs following regular stimulation and decreased with shortening S1S2 intervals. However, although RyR2(S/S) atria showed arrhythmias over a wider range of S1S2 intervals, the interatrial CV and (dV/dt)(max) of S2 APs provoking such arrhythmias were similar in RyR2(S/S) and WT. These results suggest that abnormal intracellular Ca(2+) homoeostasis produces both arrhythmic triggers and a slow-conducting arrhythmic substrate in RyR2(S/S) atria. A similar mechanism might also contribute to arrhythmogenesis in other conditions, associated with diastolic Ca(2+) release, such as atrial fibrillation.


The risk of alar necrosis associated with dermal filler injection.
Injection of dermal fillers is one of the most commonly performed cosmetic procedures. Serious complications from fillers are rare but potentially devastating to patients and physicians. Skin necrosis, such as nasal alar necrosis, is one of the most feared serious complications of dermal fillers, but there is a paucity of literature on the incidence of such events, as well as potential treatment options. We present a review of the literature and three cases of nasal alar necrosis after dermal filler injection. Nasal alar necrosis associated with dermal filler injection is a rare event. Proper technique and recognition of risk factors may reduce the incidence of this complication. Physicians should be aware of early intervention and treatment options should impending necrosis become apparent.


Soil biotic impact on plant species shoot chemistry and hyperspectral reflectance patterns.
Recent studies revealed that plant-soil biotic interactions may cause changes in above-ground plant chemistry. It would be a new step in below-ground-above-ground interaction research if such above-ground chemistry changes could be efficiently detected. Here we test how hyperspectral reflectance may be used to study such plant-soil biotic interactions in a nondestructive and rapid way. The native plant species Jacobaea vulgaris and Jacobaea erucifolius, and the exotic invader Senecio inaequidens were grown in different soil biotic conditions. Biomass, chemical content and shoot reflectance between 400 and 2500 nm wavelengths were determined. The data were analysed with multivariate statistics. Exposing the plants to soil biota enhanced the content of defence compounds. The highest increase (400%) was observed for the exotic invader S. inaequidens. Chemical and spectral data enabled plant species to be classified with an accuracy > 85%. Plants grown in different soil conditions were classified with 50-60% correctness. Our data suggest that soil microorganisms can affect plant chemistry and spectral reflectance. Further studies should test the potential to study plant-soil biotic interactions in the field. Such techniques could help to monitor, among other things, where invasive exotic plant species develop biotic resistance or the development of hotspots of crop soil diseases.


Adipokines: inflammation and the pleiotropic role of white adipose tissue.
White adipose tissue is now recognised to be a multifunctional organ; in addition to the central role of lipid storage, it has a major endocrine function secreting several hormones, notably leptin and adiponectin, and a diverse range of other protein factors. These various protein signals have been given the collective name 'adipocytokines' or 'adipokines'. However, since most are neither 'cytokines' nor 'cytokine-like', it is recommended that the term 'adipokine' be universally adopted to describe a protein that is secreted from (and synthesised by) adipocytes. It is suggested that the term is restricted to proteins secreted from adipocytes, excluding signals released only by the other cell types (such as macrophages) in adipose tissue. The adipokinome (which together with lipid moieties released, such as fatty acids and prostaglandins, constitute the secretome of fat cells) includes proteins involved in lipid metabolism, insulin sensitivity, the alternative complement system, vascular haemostasis, blood pressure regulation and angiogenesis, as well as the regulation of energy balance. In addition, there is a growing list of adipokines involved in inflammation (TNFalpha, IL-1beta, IL-6, IL-8, IL-10, transforming growth factor-beta, nerve growth factor) and the acute-phase response (plasminogen activator inhibitor-1, haptoglobin, serum amyloid A). Production of these proteins by adipose tissue is increased in obesity, and raised circulating levels of several acute-phase proteins and inflammatory cytokines has led to the view that the obese are characterised by a state of chronic low-grade inflammation, and that this links causally to insulin resistance and the metabolic syndrome. It is, however, unclear as to the extent to which adipose tissue contributes quantitatively to the elevated circulating levels of these factors in obesity and whether there is a generalised or local state of inflammation. The parsimonious view is that the increased production of inflammatory cytokines and acute-phase proteins by adipose tissue in obesity relates primarily to localised events within the expanding fat depots. It is suggested that these events reflect hypoxia in parts of the growing adipose tissue mass in advance of angiogenesis, and involve the key controller of the cellular response to hypoxia, the transcription factor hypoxia inducible factor-1.


Unmeasured Confounders in Observational Studies Comparing Bilateral Versus Single Internal Thoracic Artery for Coronary Artery Bypass Grafting: A Meta-Analysis.
Observational studies suggest a survival advantage with bilateral single internal thoracic artery (BITA) versus single internal thoracic artery grafting for coronary surgery, whereas this conclusion is not supported by randomized trials. We hypothesized that this inconsistency is attributed to unmeasured confounders intrinsic to observational studies. To test our hypothesis, we performed a meta-analysis of the observational literature comparing BITA and single internal thoracic artery, deriving incident rate ratio for mortality at end of follow-up and at 1 year. We postulated that BITA would not affect 1-year survival based on the natural history of coronary artery bypass occlusion, so that a difference between groups at 1 year could not be attributed to the intervention. We searched MEDLINE and Pubmed to identify all observational studies comparing the outcome of BITA versus single internal thoracic artery. One-year and long-term mortality for BITA and single internal thoracic artery were compared in the propensity-score-matched (PSM) series, that is, the form of observational evidence less prone to confounders. Thirty-eight observational studies (174 205 total patients) were selected for final comparison. In the 12 propensity-score-matched series (34 019 patients), the mortality reduction for BITA was similar at 1 year and at the end of follow-up (incident rate ratio, 0.70; 95% confidence interval, 0.60-0.82 versus 0.77; 95% confidence interval, 0.70-0.85; P for subgroup difference=0.43). Unmeasured confounders, rather than biological superiority, may explain the survival advantage of BITA in observational series.


Stable heterogeneity for the production of diffusible factors in cell populations.
The production of diffusible molecules that promote survival and growth is common in bacterial and eukaryotic cell populations, and can be considered a form of cooperation between cells. While evolutionary game theory shows that producers and non-producers can coexist in well-mixed populations, there is no consensus on the possibility of a stable polymorphism in spatially structured populations where the effect of the diffusible molecule extends beyond one-step neighbours. I study the dynamics of biological public goods using an evolutionary game on a lattice, taking into account two assumptions that have not been considered simultaneously in existing models: that the benefit of the diffusible molecule is a non-linear function of its concentration, and that the molecule diffuses according to a decreasing gradient. Stable coexistence of producers and non-producers is observed when the benefit of the molecule is a sigmoid function of its concentration, while strictly diminishing returns lead to coexistence only for very specific parameters and linear benefits never lead to coexistence. The shape of the diffusion gradient is largely irrelevant and can be approximated by a step function. Since the effect of a biological molecule is generally a sigmoid function of its concentration (as described by the Hill equation), linear benefits or strictly diminishing returns are not an appropriate approximations for the study of biological public goods. A stable polymorphism of producers and non-producers is in line with the predictions of evolutionary game theory and likely to be common in cell populations.


Inhaled oxytocin amplifies both vicarious reinforcement and self reinforcement in rhesus macaques (Macaca mulatta).
People attend not only to their own experiences, but also to the experiences of those around them. Such social awareness profoundly influences human behavior by enabling observational learning, as well as by motivating cooperation, charity, empathy, and spite. Oxytocin (OT), a neurosecretory hormone synthesized by hypothalamic neurons in the mammalian brain, can enhance affiliation or boost exclusion in different species in distinct contexts, belying any simple mechanistic neural model. Here we show that inhaled OT penetrates the CNS and subsequently enhances the sensitivity of rhesus macaques to rewards occurring to others as well as themselves. Roughly 2 h after inhaling OT, monkeys increased the frequency of prosocial choices associated with reward to another monkey when the alternative was to reward no one. OT also increased attention to the recipient monkey as well as the time it took to render such a decision. In contrast, within the first 2 h following inhalation, OT increased selfish choices associated with delivery of reward to self over a reward to the other monkey, without affecting attention or decision latency. Despite the differences in species typical social behavior, exogenous, inhaled OT causally promotes social donation behavior in rhesus monkeys, as it does in more egalitarian and monogamous ones, like prairie voles and humans, when there is no perceived cost to self. These findings potentially implicate shared neural mechanisms.


Long-term Effects of Pediatric Burns on the Circulatory System.
The systemic responses to burns (in particular, elevated levels of catecholamines and stress hormones) have been shown to have an impact on cardiac function for at least 3 years in children with burns. However, it is not clear if these changes lead to long-term effects on the heart. The aim of this study was to assess whether pediatric burn injury is associated with increased long-term hospital use for circulatory diseases. A population-based longitudinal study was undertaken using linked hospital and death data from Western Australia for children younger than 15 years when hospitalized for a first burn injury (n = 10 436) in 1980-2012 and a frequency matched noninjury comparison cohort, randomly selected from Western Australia's birth registrations (n = 40 819). Crude admission rates and cumulative length of stay for circulatory diseases were calculated. Negative binomial and Cox proportional hazards regression modeling were used to generate incidence rate ratios and hazard ratios, respectively. After adjustment for demographic factors and preexisting health status, the burn cohort had 1.33 (incidence rate ratio) times (95% confidence interval [CI]: 1.08-1.64) as many circulatory system hospitalizations, 2.26 times the number of days in hospital with a diagnosis of a circulatory disease (2.26, 95% CI: 1.06-4.81), and were at a higher risk of incident admissions (hazard ratio 1.22, 95% CI: 1.03-1.46), compared with the uninjured cohort. Children who sustain burn injury experience elevated hospital admission rates and increased length of hospital stay for diseases of the circulatory system for a prolonged period of time after burn discharge.


Aggregation of scaffolding protein DISC1 dysregulates phosphodiesterase 4 in Huntington's disease.
Huntington's disease (HD) is a polyglutamine (polyQ) disease caused by aberrant expansion of the polyQ tract in Huntingtin (HTT). While motor impairment mediated by polyQ-expanded HTT has been intensively studied, molecular mechanisms for nonmotor symptoms in HD, such as psychiatric manifestations, remain elusive. Here we have demonstrated that HTT forms a ternary protein complex with the scaffolding protein DISC1 and cAMP-degrading phosphodiesterase 4 (PDE4) to regulate PDE4 activity. We observed pathological cross-seeding between DISC1 and mutant HTT aggregates in the brains of HD patients as well as in a murine model that recapitulates the polyQ pathology of HD (R6/2 mice). In R6/2 mice, consequent reductions in soluble DISC1 led to dysregulation of DISC1-PDE4 complexes, aberrantly increasing the activity of PDE4. Importantly, exogenous expression of a modified DISC1, which binds to PDE4 but not mutant HTT, normalized PDE4 activity and ameliorated anhedonia in the R6/2 mice. We propose that cross-seeding of mutant HTT and DISC1 and the resultant changes in PDE4 activity may underlie the pathology of a specific subset of mental manifestations of HD, which may provide an insight into molecular signaling in mental illness in general.


An evaluation of the effects of thimerosal on neurodevelopmental disorders reported following DTP and Hib vaccines in comparison to DTPH vaccine in the United States.
Thimerosal is an ethylmercury (49.55% mercury by weight) preservative historically added to some vaccines. Toxicokinetic studies showed children in the United States received doses of mercury from Thimerosal-containing vaccines (TCVs) in excess of safety guidelines. In the United States during the 1990s, diphtheria-tetanus-pertussis (DTP) and Haemophilus influenzae type b (Hib) vaccines (maximally, 50 mug mercury per joint administration) and diphtheria-tetanus-pertussis-Haemophilus influenzae type b (DTPH) vaccines (25 mug mercury per administration) were given to children in the same childhood vaccination schedule at 2, 4, 6, and 15-18 mo, so that children receiving DTP and Hib vaccines may have maximally received an additional 100 mug more mercury exposure from TCVs than children administered DTPH vaccines. A case-control epidemiological study of neurodevelopmental disorders (NDs) reported to the Vaccine Adverse Event Reporting System (VAERS) (online public access version; updated 31 August 2004) following administration of DTP vaccines in comparison DTPH vaccines manufactured by Lederle Laboratories (Pearl River, NY) from 1994 through 1998 was undertaken. Significantly increased odds ratios for autism, speech disorders, mental retardation, infantile spasms, and thinking abnormalities reported to VAERS were found following DTP vaccines in comparison to DTPH vaccines with minimal bias or systematic error. Additional ND research should be undertaken in the context of evaluating mercury-associated exposures, especially since in 2005 the Institute of Medicine issued a report calling into question handling of vaccine safety data by the National Immunization Program of the Centers for Disease Control and Prevention.


Overexpression cdc42 attenuates isoflurane-induced neurotoxicity in developmental brain of rats.
Nowadays many children receive operations with general anesthesia. Isoflurane is a commonly-used general anesthetic. Numbers of studies demonstrated that isoflurane induced neurotoxicity and neurobehavioral deficiency in young rats, however, the underlying mechanism remained unknown. Cell division cycle 42 (cdc42) played an important role in regulating synaptic vesicle trafficking and actin dynamics in neuron, which closely linked to synaptic plasticity and dendritic spine formation. Meanwhile, cdc42 also involved in many neurodegenerative diseases. However, whether cdc42 provided a protective role in isoflurane induced synaptogenesis dysfunction still unknown. As the upstream of cdc42, calcium/Calmodulin-dependent protein kinase II (CaMKII) interacts with ion channels such as VDCCs and N-methyl-d-aspartate receptors (NMDARs), which closely associated with neuroapoptosis and cognitive deficiency in developing brain. The phosphorylation of CaMKIIα at Thr 286 plays an important role in introduction and maintenance of long-term potentiation (LTP). Therefore, we investigated the effect of isoflurane on cdc42 and its upstream Calcium/Calmodulin-dependent protein kinase II (CaMKII) and its downstream p21 activated kinase 3 (PAK3), then determined whether CaMKIIα/cdc42/PAK3 signaling pathway was involved in neurotoxicity and cognitive deficiency induced by isoflurane. Our study found that isoflurane induced neurotoxicity and resulted in cognitive impairment in young rats through suppressed CaMKIIα/cdc42/PAK3 signaling pathway. Cdc42 over-expression could reverse neurotoxicity and improve cognitive impairment induced by isoflurane.


Pharmacokinetics of low and maintenance dose valganciclovir in kidney transplant recipients.
Valganciclovir is commonly used for cytomegalovirus (CMV) prophylaxis in renal transplant patients. A fixed dose of 900 mg daily is typically recommended, however, there has never been a formal pharmacokinetic study comparing various doses in renal transplant patients. We therefore compared the pharmacokinetic characteristics of intravenous ganciclovir (IV GCV) and oral ganciclovir (GCV) with two different doses of valganciclovir (VGCV) in an open-label crossover study. Ten adult kidney recipients participated in a four-phase crossover treatment schedule of IV GCV (2.5 mg/kg every 12 h), VGCV (900 mg daily), VGCV (450 mg daily) and oral GCV (1000 mg Q8 H). IV GCV and oral VGCV 900 mg daily achieved similar values for AUC(0-24) (median 60.63 vs. 62.86 microg/h/mL). Oral VGCV 450 mg achieved comparable AUC(0-24) values as oral GCV 1000 mg Q8 H (median AUC(0-24) 35.9 vs. 29.04 microg/h/mL). Oral VGCV 900 mg daily provided systemic GCV exposure similar to IV GCV and confirms PV 16 000 study results. Further, VGCV 450 mg daily provided comparable systemic exposure versus oral GCV. Due to its favorable pharmacokinetic profile, data herein suggest that VGCV can be used in the early post-kidney transplant period, and that 450 mg daily provides ample drug exposure for effective CMV prophylaxis in kidney transplant patients.


The folding pathway of the antibody V(L) domain.
Antibodies are modular proteins consisting of domains that exhibit a beta-sandwich structure, the so-called immunoglobulin fold. Despite structural similarity, differences in folding and stability exist between different domains. In particular, the variable domain of the light chain V(L) is unusual as it is associated with misfolding diseases, including the pathologic assembly of the protein into fibrillar structures. Here, we have analysed the folding pathway of a V(L) domain with a view to determine features that may influence the relationship between productive folding and fibril formation. The V(L) domain from MAK33 (murine monoclonal antibody of the subtype kappa/IgG1) has not previously been associated with fibrillisation but is shown here to be capable of forming fibrils. The folding pathway of this V(L) domain is complex, involving two intermediates in different pathways. An obligatory early molten globule-like intermediate with secondary structure but only loose tertiary interactions is inferred. The native state can then be formed directly from this intermediate in a phase that can be accelerated by the addition of prolyl isomerases. However, an alternative pathway involving a second, more native-like intermediate is also significantly populated. Thus, the protein can reach the native state via two distinct folding pathways. Comparisons to the folding pathways of other antibody domains reveal similarities in the folding pathways; however, in detail, the folding of the V(L) domain is striking, with two intermediates populated on different branches of the folding pathway, one of which could provide an entry point for molecules diverted into the amyloid pathway.


Resolving phenotypic plasticity and species designation in the morphologically challenging Caulerpa racemosa-peltata complex (Chlorophyta, Caulerpaceae).
Although recent molecular studies have indicated the presence of a number of distinct species within the Caulerpa racemosa-peltata complex, due to the difficulties presented by high levels of phenotypic plasticity and the large number of synonyms, infra-specific taxa, and names of uncertain affinity, taxonomic proposals are yet to be made. In this study, we aimed to resolve the taxonomy of the complex and provide an example of how historical nomenclature can best be integrated into molecular based taxonomies. We accomplished this by first determining the number of genetic species within our globally sampled data set through a combination of phylogenetic and species-delimitation approaches of partial elongation factor TU and RUBISCO large subunit gene sequences. Guided by these results, comparative morphological examinations were then undertaken to gauge the extent of phenotypic plasticity within each species, as well as any morphological overlap between them. Our results revealed the presence of 11 distinct species within the complex, five of which showed high levels of phenotypic plasticity and partial overlap with other species. On the basis of observations of a large number of specimens, including type specimens/descriptions, and geographic inferences, we were able to confidently designate names for the lineages. Caulerpa peltata, C. imbricata and C. racemosa vars. laetevirens, occidentalis and turbinata were found to represent environmentally induced forms of a single species, for which the earlier-described C. chemnitzia, previously regarded as a synonym of C. racemosa var. turbinata, is reinstated. C. cylindracea, C. lamourouxii, C. macrodisca, C. nummularia and C. oligophylla are also reinstated and two new species, C. macra stat. nov. and C. megadisca sp. nov., are proposed.


Appraisal of salinity and fluoride in a semi-arid region of India using statistical and multivariate techniques.
Various physico-chemical parameters, including fluoride (F(-)), were analyzed to understand the hydro-geochemistry of an aquifer in a semi-arid region of India. Furthermore, the quality of the shallow and deep aquifer (using tube well and hand pumps) was also investigated for their best ecological use including drinking, domestic, agricultural and other activities. Different multivariate techniques were applied to understand the groundwater chemistry of the aquifer. Findings of the correlation matrix were strengthened by the factor analysis, and this shows that salinity is mainly caused by magnesium salts as compared to calcium salts in the aquifer. The problem of salinization seems mainly compounded by the contamination of the shallow aquifers by the recharging water. High factor loading of total alkalinity and bicarbonates indicates that total alkalinity was mainly due to carbonates and bicarbonates of sodium. The concentration of F(-) was found more in the deep aquifer than the shallow aquifer. Further, only a few groundwater samples lie below the permissible limit of F(-), and this indicates a risk of dental caries in the populace of the study area. The present study indicates that regular monitoring of groundwater is an important step to avoid human health risks and to assess its quality for various ecological purposes.


Prediction of polycyclic aromatic hydrocarbon biodegradation in contaminated soils using an aqueous hydroxypropyl-beta-cyclodextrin extraction technique.
This study investigated the use of an aqueous hydroxypropyl-beta-cyclodextrin (HPCD) shake extraction to predict the degree of microbial degradation of polycyclic aromatic hydrocarbons (PAHs) in soils. Three different aged PAH-contaminated soils were studied: A soil from a former coke works (CW) and two artificially contaminated soils (AC1 and AC2). First, the catabolic activity of the indigenous soil microflora was assessed with 14C-respirometry, using a range of 14C-labeled aromatic compounds. Extensive mineralization of several compounds occurred in the CW and the AC2 soils, suggesting that both soils contained catabolically active microorganisms. No significant mineralization occurred in the AC1 soil, implying that either it did not contain an indigenous PAH-degrading microbial population or that degradation, but not mineralization, occurred. The soils then were subjected to three sets of analyses: dichloromethane (DCM) soxhlet extraction, six-week biodegradation assay followed by DCM extraction, and extraction with HPCD followed by DCM extraction. A general decrease in PAHs present in the soils occurred after the biodegradation assay. In the CW and the AC1 soils, strong correlations were observed between the amount of PAHs biodegraded and the fraction of PAHs removed from the soils using the HPCD extraction. However, the AC2 soil showed a more modest correlation between the biodegradable fraction and the HPCD extractable fraction, with the HPCD extraction slightly underestimating the extent of PAH biodegradation. The results of this study indicated that an aqueous HPCD extraction may be a useful tool in assessing the microbial availability of aged contaminant mixtures in soils, although further validation is required.


Reproductive Coercion and Intimate Partner Violence Victimization Among a Racially and Ethnically Diverse Young Adult Sample.
Reproductive coercion is an understudied form of intimate partner violence (IPV) that occurs when a person attempts to control the autonomous reproductive decision making of their intimate partner. Previous research has demonstrated that individuals who experience reproductive coercion are more likely to experience other forms of IPV (e.g., physical and sexual). Research has also shown that Black/African American and Latinx/Hispanic individuals are at an increased risk of experiencing reproductive coercion compared to their White/Non-Hispanic peers. However, most of the research on reproductive coercion has been conducted in family-planning clinics where IPV is reported at a higher rate than in community samples. Thus, using a diverse community sample of female-identifying young adults (N = 370) who were recruited as part of an ongoing longitudinal study on dating violence, we examined the prevalence of lifetime reproductive coercion and its relationship with other forms of IPV, as well as the differences in prevalence among racial and ethnic groups. Lifetime prevalence of being victimized by any form of reproductive coercion was 11.4%. Results indicated that individuals who experienced reproductive coercion were more likely to experience physical and sexual IPV relative to those who did not experience reproductive coercion. With respect to race/ethnicity, 5.6% of White participants, 10.5% of Black/African American participants, and 14.8% of Hispanic/Latinx participants reported experiencing reproductive coercion. Chi-square analyses showed Hispanic/Latinx participants had a significantly higher prevalence of reproductive coercion when compared to White/Non-Hispanic participants. These findings suggest a need for additional research on culturally-specific risk and protective factors related to reproductive coercion among Hispanic/Latinx individuals to identify potential intervention and prevention strategies.


Prospective study of the accuracy of the surgeon's diagnosis in 2000 excised skin tumors.
Expeditious yet efficacious removal of skin tumors is a common responsibility for the plastic surgeon. The need to minimize potential risks for mortality or morbidity from undue or excessive surgical resections and to control costs by avoiding unnecessary procedures behooves us to make a precise clinical diagnosis preceding any decision even for such "minor" surgery. Just how accurate these decisions can be expected to be for a typical surgical practice was scrutinized by means of this prospective 4-year study involving the resection of 2058 skin lesions. Each lesion was initially assigned a clinical diagnosis after a brief gross examination and then compared with the pathology report, which was always considered to be the correct answer. Within these parameters, only 65 percent of all tumors were identified correctly preoperatively. Two-thirds of all lesions were benign. Three-quarters of benign lesions were as assumed, and 92 percent of all presumed benign lesions were benign even if incorrectly identified initially, whereas fortunately only 3 percent proved to be malignant. On the other hand, only three-fifths of malignant lesions were identified correctly clinically, yet only 11 percent were benign, implying that most such lesions properly deserved excision anyway. Therefore, approximately 90 percent of all lesions whether benign or malignant were removed appropriately without compromising the patient, but to expect a clinical acumen of 100 percent in this setting may not be realistic. The accuracy of the surgeon in identifying lesions as probably benign was certainly high enough that cost-containment mechanisms designed to deny authorization for their removal probably would be justifiable and difficult to appeal. Any suspicious or equivocal lesions still will require mandatory intervention despite such constraints, because often only histologic examination will allow a definitive diagnosis.


Effects on adult cognitive function after neonatal exposure to clinically relevant doses of ionising radiation and ketamine in mice.
Radiological methods for screening, diagnostics and therapy are frequently used in healthcare. In infants and children, anaesthesia/sedation is often used in these situations to relieve the patients' perception of stress or pain. Both ionising radiation (IR) and ketamine have been shown to induce developmental neurotoxic effects and this study aimed to identify the combined effects of these in a murine model. Male mice were exposed to a single dose of ketamine (7.5 mg kg-1 body weight) s.c. on postnatal day 10. One hour after ketamine exposure, mice were whole body irradiated with 50-200 mGy gamma radiation (137Cs). Behavioural observations were performed at 2, 4 and 5 months of age. At 6 months of age, cerebral cortex and hippocampus tissue were analysed for neuroprotein levels. Animals co-exposed to IR and ketamine displayed significant (P≤0.01) lack of habituation in the spontaneous behaviour test, when compared with controls and single agent exposed mice. In the Morris Water Maze test, co-exposed animals showed significant (P≤0.05) impaired learning and memory capacity in both the spatial acquisition task and the relearning test compared with controls and single agent exposed mice. Furthermore, in co-exposed mice a significantly (P≤0.05) elevated level of tau protein in cerebral cortex was observed. Single agent exposure did not cause any significant effects on the investigated endpoints. Co-exposure to IR and ketamine can aggravate developmental neurotoxic effects at doses where the single agent exposure does not impact on the measured variables. These findings show that estimation of risk after paediatric low-dose IR exposure, based upon radiation dose alone, may underestimate the consequences for this vulnerable population.


Introducing a Favourite Tip Definition and Projection with Tripod Suture in Rhinoplasty.
Surgical manipulation of the lower lateral cartilages of nasal tip could cause subtle but significant responses to the manipulation. The suture-techniques almost always offer a reliable and dramatic method of tip reshaping without a need to break or interrupt the alar rim strip or even adding tip graft. The aim of this study was to describe and assess outcomes of a new suture technique in rhinoplasty by using cephalic dome septal rotation suture for better tip definition. All consecutive women (62 women) who attended our centre with nasal tip drooping and some degree of columellar retraction, underwent this technique from January 2005 to September 2009. Their ages ranged from 17 to 32 years old. Fifty one primary open and 11 secondary open rhinoplasties were performed using the conventional techniques and then the new suture technique was applied before closing the incisions on each patient. The patients were followed-up from 22 to 72 months (mean = 52.3). The projection and rotation was satisfactory in all patients and the final results persisted with only minimal changes over time. It is important to say that this new suture is different from 'tip rotation suture' that was described by Tebbets. Our suture maintains the favorite tip definition with slight dorsal tip inclination. Indeed, it causes a two-point definition as the columellar lobular angle and the supra tip break definition. In addition, long-term follow-up has shown that this new suture (i.e., cephalic dome-septal rotation suture) is able to maintain optimal position of the nasal tip projection and definition.


Comparison of two automatic cell-counting solutions for fluorescent microscopic images.
Cell counting in microscopic images is one of the fundamental analysis tools in life sciences, but is usually tedious, time consuming and prone to human error. Several programs for automatic cell counting have been developed so far, but most of them demand additional training or data input from the user. Most of them do not allow the users to online monitor the counting results, either. Therefore, we designed two straightforward, simple-to-use cell-counting programs that also allow users to correct the detection results. In this paper, we present the Cellcounter and Learn123 programs for automatic and semiautomatic counting of objects in fluorescent microscopic images (cells or cell nuclei) with a user-friendly interface. Although Cellcounter is based on predefined and fine-tuned set of filters optimized on sets of chosen experiments, Learn123 uses an evolutionary algorithm to determine the adapt filter parameters based on a learning set of images. Cellcounter also includes an extension for analysis of overlaying images. The efficiency of both programs was assessed on images of cells stained with different fluorescent dyes by comparing automatically obtained results with results that were manually annotated by an expert. With both programs, the correlation between automatic and manual counting was very high (R(2) < 0.9), although Cellcounter had some difficulties processing images with no cells or weakly stained cells, where sometimes the background noise was recognized as an object of interest. Nevertheless, the differences between manual and automatic counting were small compared to variations between experimental repeats. Both programs significantly reduced the time required to process the acquired images from hours to minutes. The programs enable consistent, robust, fast and accurate detection of fluorescent objects and can therefore be applied to a range of different applications in different fields of life sciences where fluorescent labelling is used for quantification of various phenomena. Moreover, Cellcounter overlay extension also enables fast analysis of related images that would otherwise require image merging for accurate analysis, whereas Learn123's evolutionary algorithm can adapt counting parameters to specific sets of images of different experimental settings.


The Duffy antigen receptor for chemokines transports chemokines and supports their promigratory activity.
The Duffy antigen receptor for chemokines (DARC) belongs to a family of 'silent' heptahelical chemokine receptors that do not couple to G proteins and fail to transmit measurable intracellular signals. DARC binds most inflammatory chemokines and is prominently expressed on venular endothelial cells, where its function has remained contentious. Here we show that DARC, like other silent receptors, internalized chemokines but did not effectively scavenge them. Instead, DARC mediated chemokine transcytosis, which led to apical retention of intact chemokines and more leukocyte migration across monolayers expressing DARC. Mice overexpressing DARC on blood vessel endothelium had enhanced chemokine-induced leukocyte extravasation and contact-hypersensitivity reactions. Thus, interactions of chemokines with DARC support their activity on apposing leukocytes in vitro and in vivo.


Risk factors for early failure of surgical amputations: an analysis of 8,878 isolated lower extremity amputation procedures.
There are very few data currently published on risk factors for early failure of lower extremity amputation procedures. All patients from the 2005-2010 American College of Surgeons NSQIP database who underwent isolated lower extremity amputation were included for analysis (excluding patients with earlier operation within 30 days, patients undergoing an open amputation, and patients undergoing another procedure during amputation). Multivariate logistic regression was used to determine predictors of early amputation failure (defined as need for reoperation within 30 days postoperatively) after adjustment for a number of preoperative and intraoperative variables. A total of 8,878 patients were included for analysis (4,258 below-knee amputations [BKA]; 3,415 above-knee amputations; and 1,205 transmetatarsal amputations). Overall rate of early amputation failure was 12.7% (12.6% for BKA, 8.1% for above-knee amputations, and 26.4% for transmetatarsal amputations; p < 0.0001). Several pre- and intraoperative variables appeared to be independently associated with early amputation failure, including emergency operation, transmetatarsal amputation (reference = BKA), sepsis (reference = no sepsis), septic shock (reference = no sepsis), end-stage renal disease, systemic inflammatory response syndrome (reference = no sepsis), intraoperative surgical trainee participation, body mass index ≥30, and ongoing tobacco use. Characteristics associated with decreased early amputation failure include age 80 years or older (reference = younger than 65 years), locoregional anesthesia, above-knee amputation (reference = BKA), operative time 40 to 59 minutes (reference = <40 minutes), operative time ≥80 minutes (reference = <40 minutes), and operative time 60 to 79 minutes (reference = <40 minutes). Increased operative time and heightened supervision of participating surgical trainees can decrease the risk of early amputation failure. In addition, specific clinical situations, such as sepsis or emergency procedures, should prompt vascular surgeons to consider either an open amputation procedure or a more proximal closed amputation.


Evaluation of Cystatin C as a marker of renal injury following on-pump and off-pump coronary surgery.
Cardiopulmonary bypass is regarded as an important contributor to renal injury, whereas off-pump surgery is considered less damaging. Cystatin C, a cystine protease inhibitor, is more sensitive and specific than creatinine in the assessment of renal function. We assessed the value of Cystatin C in quantifying clinical and subclinical renal injury following on-pump and off-pump cardiac surgery. Sixty consecutive patients were recruited: 30 patients undergoing on-pump CABG and 30 patients undergoing off-pump CABG. Blood samples were collected pre-operatively and on days 1, 2 and 4 postoperatively to measure serum creatinine and serum Cystatin C. Urinary samples were collected concurrently to measure microalbumin and N-acetyl-beta-glucosaminidase, denoting changes in renal glomerular and tubular function respectively. The off-pump group were older (65+/-11 vs. 61+/-8 years; P=0.046), had a higher mean Parsonnet score (9.4+/-6.2 vs. 5.4+/-3.6, P<0.01) and received fewer grafts (2.4+/-0.9 vs. 3.1+/-0.7; P<0.01) compared to the on-pump group. The groups were otherwise matched with respect to preoperative renal function and left ventricular function. Patients undergoing on-pump CABG had greater increases in all four parameters of renal injury. Adjustment for preoperative differences increased the size and significance of the effect of off-pump vs. on-pump surgery (percent difference 13%; 95% CI 2-26 for creatinine, and 16%; 95% CI 4-29 for Cystatin C). Cystatin and creatinine were strongly correlated with each other. Avoidance of cardiopulmonary bypass may reduce renal injury particularly in higher risk patients. Cystatin C proved to be a simple and sensitive measure of overall renal function and can be used in the routine assessment of renoprotective strategies.


A case with oto-spondylo-mega-epiphyseal-dysplasia (OSMED): the clinical recognition and differential diagnosis.
The oto-spondylo-mega-epiphyseal-dysplasia (OSMED) phenotype is an autosomal recessive trait that is a skeletal dysplasia with the hallmark findings of limb shortening, multiple skeletal and radiological abnormalities, mid-face hypoplasia with a flat nasal bridge, small upturned nasal tip, and sensorineural hearing loss. A 3.5-year-old girl born to consanguineous Turkish parents had characteristic facial features at birth: mid-face hypoplasia, mild hypertelorism, upslanting palpebral fissures, prominent supraorbital ridges, depressed nasal bridge, small upturned nasal tip, long philtrum, and micrognathia. Radiological examination at three years of age revealed large flaring metaphyses and wide flat epiphyses. The humerus and femur showed the characteristic dumbbell shape. She had bilateral hearing loss with no ophthalmologic findings. There is continuing debate over the clinical overlap and differential diagnosis of OSMED syndrome. The patient was examined considering Weissenbacher-Zweymuller, Stickler type 3, Marshall syndrome, and Kniest dysplasia as possible differential diagnoses. We believe that the presented patient clinically manifested features of OSMED syndrome. We would like to point out that the management of OSMED calls for a coordinated multidisciplinary approach.


Relationship Between P15 Gene Mutation and Formation and Metastasis of Malignant Osteosarcoma.
BACKGROUND As a type of primary malignant bone tumor, osteosarcoma has high incidence and poor prognosis, and is predisposed for pulmonary metastasis. The abnormal expression of P15 gene directly participates in the invasion of various cancers. Therefore, this study investigated the gene mutation of P15 in both primary lesion and pulmonary metastasis lesion of osteosarcoma in a rat model, in an attempt to elucidate the value of P15 gene as a biological marker. MATERIAL AND METHODS A total of 60 SD rats were randomly divided into 2 groups. Model rats had injection of osteosarcoma UMR-106 cells (5×106) inoculated underneath the right forelimb skin, while control rats received saline injection instead. Six rats were sacrificed after 0, 1, 2, 4, and 6 weeks of the inoculation. Tissue samples from inoculation sites and lungs were extracted for measuring the tumor size. SP immunohistochemical (IHC) staining was used to detect the positive expression rate, while P15 gene mutation was detected by PCR method. RESULTS With the elongation of inoculation time, tumor size was significantly increased (p<0.05). The positive expression rates in both primary and pulmonary metastasis lesions were also significantly elevated (p<0.05). The occurrence rate of P15 gene mutation in model rats was significantly elevated and showed a correlation with the tumor formation (r=0.998, p<0.05). CONCLUSIONS The P15 gene mutation was significantly correlated with osteosarcoma formation and metastasis towards the pulmonary tissue, suggesting its potency as a novel biological marker for early diagnosis of osteosarcoma.


Ca2+/calmodulin-dependent protein kinase II and protein kinase C phosphorylate a synthetic peptide corresponding to a sequence that is specific for the gamma 2L subunit of the GABAA receptor.
The gamma 2 subunit of the GABA receptor (GABAA-R) is alternatively spliced. The long variant (gamma 2L) contains eight additional amino acids that possess a consensus sequence site for protein phosphorylation. Previous studies have demonstrated that a peptide or fusion protein containing these eight amino acids is a substrate for protein kinase C (PKC), but not cyclic AMP-dependent protein kinase A (PKA)-stimulated phosphorylation. We have examined the ability of PKA, PKC, and Ca2+/calmodulin-dependent protein kinase (CAM kinase II) to phosphorylate a synthetic peptide corresponding to residues 336-351 of the intracellular loop of the gamma 2L subunit and inclusive of the alternatively spliced phosphorylation consensus sequence site. PKC and CAM kinase II produced significant phosphorylation of this peptide, but PKA was ineffective. The Km values for PKC- and CAM kinase II-stimulated phosphorylation of this peptide were 102 and 35 microM, respectively. Maximal velocities of 678 and 278 nmol of phosphate/min/mg were achieved by PKC and CAM kinase II, respectively. The phosphorylation site in the eight-amino-acid insert of the gamma 2L subunit has been shown to be necessary for ethanol potentiation of the GABAA-R. Thus, our results suggest that PKC, CAM kinase II, or both may play a role in the effects of ethanol on GABAergic function.


Can Unconventional Immunomodulatory Agents Help Alleviate COVID-19 Symptoms and Severity?
Severe acute respiratory syndrome coronavirus 2 (SARS coronavirus 2, or SARS-CoV-2) is the cause of the respiratory infection known as COVID-19. From an immunopathological standpoint, coronaviruses such as SARS-CoV-2 induce increased levels of a variety of T-helper 1 (Th1) and inflammatory cytokines and chemokines, including interleukin-1 (IL-1), IL-6, CCL2 protein, and CXCL10 protein. In the absence of proven antiviral agents or an effective vaccine, substances with immunomodulatory activity may be able to inhibit inflammatory and Th1 cytokines and/or yield an anti-inflammatory and/or Th2 immune response to counteract COVID-19 symptoms and severity. This report briefly describes the following four unconventional but commercially accessible immunomodulatory agents that can be employed in clinical trials to evaluate their effectiveness at alleviating disease symptoms and severity: low-dose oral interferon alpha, microdose DNA, low-dose thimerosal, and phytocannabinoids.


Pure non-dioxin-like PCB congeners suppress induction of AhR-dependent endpoints in rat liver cells.
The relative potencies of non-ortho-substituted coplanar polychlorinated biphenyl (PCB) congeners to activate the aryl hydrocarbon receptor (AhR) and to cause the AhR-dependent toxic events are essential for their risk assessment. Since some studies suggested that abundant non-dioxin-like PCB congeners (NDL-PCBs) may alter the AhR activation by PCB mixtures and possibly cause non-additive effects, we evaluated potential suppressive effects of NDL-PCBs on AhR activation, using a series of 24 highly purified NDL-PCBs. We investigated their impact on the model AhR agonist-induced luciferase reporter gene expression in rat hepatoma cells and on induction of CYP1A1/1B1 mRNAs and deregulation of AhR-dependent cell proliferation in rat liver epithelial cells. PCBs 128, 138, and 170 significantly suppressed AhR activation (with IC50 values from 1.4 to 5.6 μM), followed by PCBs 28, 47, 52, and 180; additionally, PCBs 122, 153, and 168 showed low but still significant potency to reduce luciferase activity. Detection of CYP1A1 mRNA levels in liver epithelial cells largely confirmed these results for the most abundant NDL-PCBs, whereas the other AhR-dependent events (CYP1B1 mRNA expression, induction of cell proliferation in confluent cells) were less sensitive to NDL-PCBs, thus indicating a more complex regulation of these endpoints. The present data suggest that some NDL-PCBs could modulate overall dioxin-like effects in complex mixtures.


Analysis of Induction and Establishment of Dwarf Bunt of Wheat Under Marginal Climatic Conditions.
Dwarf bunt caused by Tilletia contraversa is a disease of winter wheat that has a limited geographic distribution due to specific winter climate requirements. The pathogen is listed as a quarantine organism by several countries that may have wheat production areas with inadequate or marginal climate for the disease-in particular the People's Republic of China. Field experiments were conducted in the United States in an area of Kansas that is a climatic analog to the northern winter wheat areas of China to evaluate the risk of disease introduction into such areas. The soil surface of four replicate 2.8 × 9.75 m plots, planted with a highly susceptible cultivar, was inoculated with six teliospore concentrations ranging from 0.88 to 88,400 teliospores/cm2. A single initial inoculation was done in each of three nurseries planted during separate seasons followed by examination for disease for 4 to 6 years afterward. Any diseased spikes produced were crushed and returned to the plots where they were produced. One nursery had no disease during all six seasons. In two nurseries, the disease was induced at trace levels at the three highest inoculation rates. Disease carryover to the second year occurred during one year in one nursery in plots at the highest inoculation rate, but no disease occurred the following three seasons. A duplicate nursery planted in a disease conducive area in Utah demonstrated that the highest rate of inoculum used in the experiments was sufficient to cause almost 100% infection. This study demonstrated that in an area with marginal climatic conditions it was possible to induce transient trace levels of dwarf bunt, but the disease was not established even with a highly susceptible cultivar and high levels of inoculum. Our results support the conclusions of the 1999 Agreement on U.S.-China Agricultural Cooperation which set a tolerance for teliospores in grain, and supports the Risk Assessment Model for Importation of United States Milling Wheat Containing T. contraversa.


Transperitoneal versus retroperitoneal laparoscopic adrenalectomy for adrenal tumours in adults.
Laparoscopic adrenalectomy is an accepted treatment worldwide for adrenal gland disease in adults. The transperitoneal approach is more common. The retroperitoneal approach may be preferred, to avoid entering the peritoneum, but no clear advantage has been demonstrated so far. To assess the effects of laparoscopic transperitoneal adrenalectomy (LTPA) versus laparoscopic retroperitoneal adrenalectomy (LRPA) for adrenal tumours in adults. We searched CENTRAL, MEDLINE, Embase, ICTRP Search Portal, and ClinicalTrials.gov to 3 April 2018. We applied no language restrictions. Two review authors independently scanned the abstract, title, or both sections of every record retrieved to identify randomised controlled trials (RCTs) on laparoscopic adrenalectomy for preoperatively assessed adrenal tumours. Participants were affected by corticoid and medullary, benign and malignant, functional and silent tumours or masses of the adrenal gland, which were assessed by both laboratory and imaging studies. Two review authors independently extracted data, assessed trials for risk of bias, and evaluated overall study quality using GRADE criteria. We calculated the risk ratio (RR) for dichotomous outcomes, or the mean difference (MD) for continuous variables, and corresponding 95% confidence interval (CI). We primarily used a random-effects model for pooling data. We examined 1069 publications, scrutinized 42 full-text publications or records, and included five RCTs. Altogether, 244 participants entered the five trials; 127 participants were randomised to retroperitoneal adrenalectomy and 117 participants to transperitoneal adrenalectomy. Two trials had a follow-up of nine months, and three trials a follow-up of 31 to 70 months. Most participants were women, and the average age was around 40 years. Three trials reported all-cause mortality; in two trials, there were no deaths, and in one trial with six years of follow-up, four participants died in the LRPA group and one participant in the LTPA group (164 participants; low-certainty evidence). The trials did not report all-cause morbidity. Therefore, we analysed early and late morbidity, and included specific adverse events under these outcome measures. The results were inconclusive between LRPA and LTPA for early morbidity (usually reported within 30 to 60 days after surgery; RR 0.56, 95% CI 0.27 to 1.16; P = 0.12; 5 trials, 244 participants; very low-certainty evidence). Nine out of 127 participants (7.1%) in the LRPA group, compared with 16 out of 117 participants (13.7%) in the LTPA group experienced an adverse event. Participants in the LRPA group may have a lower risk of developing late morbidity (reported as latest available follow-up; RR 0.12, 95% CI 0.01 to 0.92; P = 0.04; 3 trials, 146 participants; very low-quality evidence). None of the 78 participants in the LRPA group, compared with 7 of the 68 participants (10.3%) in the LTPA group experienced an adverse event.None of the trials reported health-related quality of life. The results were inconclusive for socioeconomic effects, assessed as time to return to normal activities and length of hospital stay, between the intervention and comparator groups (very low-certainty evidence). Participants who had LRPA may have had an earlier start on oral fluid or food intake (MD -8.6 hr, 95% CI -13.5 to -3.7; P = 0.0006; 2 trials, 89 participants), and ambulation (MD -5.4 hr, 95% CI -6.8 to -4.0 hr; P < 0.0001; 2 trials, 89 participants) than those in the LTPA groups. Postoperative and operative parameters (duration of surgery, operative blood loss, conversion to open surgery) showed inconclusive results between the intervention and comparator groups. The body of evidence on laparoscopic retroperitoneal adrenalectomy compared with laparoscopic transperitoneal adrenalectomy is limited. Late morbidity might be reduced following laparoscopic retroperitoneal adrenalectomy, but we are uncertain about this effect because of very low-quality evidence. The effects on other key outcomes, such as all-cause mortality, early morbidity, socioeconomic effects, and operative and postoperative parameters are uncertain. LRPA might show a shorter time to oral fluid or food intake and time to ambulation, but we are uncertain whether this finding can be replicated. New long-term RCTs investigating additional data, such as health-related quality of life, surgeons' level of experience, treatment volume of surgical centres, and details on techniques used are needed.


Population attributable risk of unintentional childhood poisoning in Karachi Pakistan.
The percentage of unintentional childhood poisoning cases in a given population attributable to specific risk factors (i.e., the population attributable risk) which can be calculated; determination of such risk factors associated with potentially modifiable risk factors, are necessary to focus on the prevention strategies. We calculated PARs, using 120 cases with unintentional poisoning and 360 controls in a hospital based matched case- control study. The risk factors were accessibility to hazardous chemicals and medicines due to unsafe storage, child behavior reported as hyperactive, storage of kerosene and petroleum in soft drink bottles, low socioeconomic class, less education of the mother and the history of previous poisoning. THE FOLLOWING ATTRIBUTED RISKS WERE OBSERVED: 12% (95% confidence interval [CI] = 8%-16%) for both chemicals and medicines stored unsafe, 19% (15%-23%) for child reported as hyperactive, 40% (38%-42%) for storage of kerosene and petroleum in soft drink bottles, 48% (42%-54%) for low socioeconomic status, 38% (32%-42%) for no formal mothers education and 5.8% (2%-10%) for history of previous poisoning. 48% of cases for overall study population which could be attributed to at least one of the six risk factors. Among girls, this proportion was 23% and 43% among boys. About half of the unintentional childhood poisoning cases in this Pakistani population could be avoided. Exposure to potentially modifiable risk indicators explained about half of the cases of unintentional poisoning among children under five years of age in this Pakistani population, indicating the theoretical scope for prevention of the disease.


Antiproliferative effects of dibenzocyclooctadiene lignans isolated from Schisandra chinensis in human cancer cells.
Dibenzocyclooctadiene lignans isolated from Schisandra chinensis showed antiproliferative effects in various human cancer cells. The methoxy groups at C-3, C-4, C-3', and C-4', the hydroxyl group at C-8', and the stereo-configuration of the biphenyl ring and the angeloyl group might have influence on these activities. Additional studies indicate that one of mechanism of action of an active compound schizantherin C in A549 human lung cancer cells was related to the inhibition of cell cycle progression in G0/G1 phase.


Effect of CYP3A5 genotypes on the pharmacokinetics of carbamazepine when used as monotherapy or co-administered with phenytoin, phenobarbital or valproic acid in Thai patients.
To determine the effects of CYP3A5 polymorphisms on carbamazepine (CBZ) pharmacokinetic parameters when CBZ is used either as monotherapy or co-administered with phenytoin (PHT), phenobarbital (PB) or valproic acid (VPA). Retrospective data were collected from an electronic database and medical records. Blood samples were obtained and drug concentrations analyzed as a part of routine therapeutic drug monitoring (TDM). Screening for wild-type CYP3A5*1 and CYP3A5*3 single nucleotide polymorphism (rs776746) by allelic discrimination assay using real-time polymerase chain reaction technique (real-time PCR) was performed. Pharmacokinetic parameters of CBZ; clearance and dose-adjusted CBZ levels in patients with different genotypes were calculated and compared. Of the 70 patients assessed, 8 (11%) patients were homozygous CYP3A5*1/*1, 28 (40%) patients were heterozygous CYP3A5*1/*3, and 34 (49%) patients were homozygous CYP3A5*3/*3. The CBZ clearance and dose-adjusted CBZ levels did not significantly differ between patients with CYP3A5*1 and CYP3A5*3 alleles when CBZ was used as monotherapy. For patients who used CBZ in combination with an enzyme-inducing antiepileptic drug (AED: PHT or PB), individuals carrying the CYP3A5*1 allele (CYP3A5 expressers) showed a trend of having higher CBZ clearance and lower dose-adjusted CBZ level as compared to individuals carrying the CYP3A5*3 allele, even though no statistical significance was recorded. Nevertheless, it was observed that AEDs significantly increased CBZ clearance only in patients carrying the active CYP3A5*1 allele. When CBZ was used in combination with enzyme-inducing AED, CYP3A5 expressers yielded a trend toward greater susceptibility to change in CBZ clearance and showed lower dose-adjusted CBZ levels compared to CYP3A5 non-expressers. The dosage regimen should be adjusted accordingly to gain a better clinical outcome.This article is open to POST-PUBLICATION REVIEW. Registered readers (see "For Readers") may comment by clicking on ABSTRACT on the issue's contents page.


Leukotriene E4 activates human Th2 cells for exaggerated proinflammatory cytokine production in response to prostaglandin D2.
PGD(2) exerts a number of proinflammatory responses through a high-affinity interaction with chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2) and has been detected at high concentrations at sites of allergic inflammation. Because cysteinyl leukotrienes (cysLTs) are also produced during the allergic response, we investigated the possibility that cysLTs may modulate the response of human Th2 cells to PGD(2). PGD(2) induced concentration-dependent Th2 cytokine production in the absence of TCR stimulation. Leukotrienes D(4) and E(4) (LTE(4)) also stimulated the cytokine production but were much less active than PGD(2). However, when combined with PGD(2), cysLTs caused a greater than additive enhancement of the response, with LTE(4) being most effective in activating Th2 cells. LTE(4) enhanced calcium mobilization in response to PGD(2) in Th2 cells without affecting endogenous PGD(2) production or CRTH2 receptor expression. The effect of LTE(4) was inhibited by montelukast but not by the P2Y(12) antagonist methylthioadenosine 5'-monophosphate. The enhancing effect was also evident with endogenous cysLTs produced from immunologically activated mast cells because inhibition of cysLT action by montelukast or cysLT synthesis by MK886, an inhibitor of 5-lipoxygenase-activating protein, reduced the response of Th2 cells to the levels produced by PGD(2) alone. These findings reveal that cysLTs, in particular LTE(4), have a significant proinflammatory impact on T cells and demonstrate their effects on Th2 cells are mediated by a montelukast-sensitive receptor.


Neural network method to correct bidirectional effects in water-leaving radiance.
Ocean color algorithms that rely on "atmospherically corrected" nadir water-leaving radiances to infer information about marine constituents such as the chlorophyll concentration depend on a reliable method to convert the angle-dependent measured radiances from the observation direction to the nadir direction. It is also important to convert the measured radiances to the nadir direction when comparing and merging products from different satellite missions. The standard correction method developed by Morel and coworkers requires knowledge of the chlorophyll concentration. Also, the standard method was developed based on the Case 1 (open ocean) assumption, which makes it unsuitable for Case 2 situations such as turbid coastal waters. We introduce a neural network method to convert the angle-dependent water-leaving radiance (or the corresponding remote sensing reflectance) from the observation direction to the nadir direction. This method relies on neither an "atmospheric correction" nor prior knowledge of the water constituents or the inherent optical properties. It directly converts the remote sensing reflectance from an arbitrary slanted viewing direction to the nadir direction by using a trained neural network. This method is fast and accurate, and it can be easily adapted to different remote sensing instruments. Validation using NuRADS measurements in different types of water shows that this method is suitable for both Case 1 and Case 2 waters. In Case 1 or chlorophyll-dominated waters, our neural network method produces corrections similar to those of the standard method. In Case 2 waters, especially sediment-dominated waters, a significant improvement was obtained compared to the standard method.


An ARHGAP25 variant links aberrant Rac1 function to early-onset skeletal fragility.
Ras homologous guanosine triphosphatases (RhoGTPases) control several cellular functions, including cytoskeletal actin remodeling and cell migration. Their activities are downregulated by GTPase-activating proteins (GAPs). Although RhoGTPases are implicated in bone remodeling and osteoclast and osteoblast function, their significance in human bone health and disease remains elusive. Here, we report defective RhoGTPase regulation as a cause of severe, early-onset, autosomal-dominant skeletal fragility in a three-generation Finnish family. Affected individuals (n = 13) presented with multiple low-energy peripheral and vertebral fractures despite normal bone mineral density (BMD). Bone histomorphometry suggested reduced bone volume, low surface area covered by osteoblasts and osteoclasts, and low bone turnover. Exome sequencing identified a novel heterozygous missense variant c.652G>A (p.G218R) in ARHGAP25, encoding a GAP for Rho-family GTPase Rac1. Variants in the ARHGAP25 5' untranslated region (UTR) also associated with BMD and fracture risk in the general population, across multiple genomewide association study (GWAS) meta-analyses (lead variant rs10048745). ARHGAP25 messenger RNA (mRNA) was expressed in macrophage colony-stimulating factor (M-CSF)-stimulated human monocytes and mouse osteoblasts, indicating a possible role for ARHGAP25 in osteoclast and osteoblast differentiation and activity. Studies on subject-derived osteoclasts from peripheral blood mononuclear cells did not reveal robust defects in mature osteoclast formation or resorptive activity. However, analysis of osteosarcoma cells overexpressing the ARHGAP25 G218R-mutant, combined with structural modeling, confirmed that the mutant protein had decreased GAP-activity against Rac1, resulting in elevated Rac1 activity, increased cell spreading, and membrane ruffling. Our findings indicate that mutated ARHGAP25 causes aberrant Rac1 function and consequently abnormal bone metabolism, highlighting the importance of RhoGAP signaling in bone metabolism in familial forms of skeletal fragility and in the general population, and expanding our understanding of the molecular pathways underlying skeletal fragility. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.


Greece: Health system review.
The Health Systems in Transition (HiT) profiles are country-based reports that provide a detailed description of a health system and of policy initiatives in progress or under development. HiTs examine different approaches to the organization, financing and delivery of health services and the role of the main actors in health systems; describe the institutional framework, process, content and implementation of health and health care policies; and highlight challenges and areas that require more in-depth analysis. The health status of the Greek population has strongly improved over the last few decades and seems to compare relatively favourably with other OECD and European Union (EU) countries. The health system is a mixture of public integrated, public contract and public reimbursement models, comprising elements from both the public and private sectors and incorporating principles of different organizational patterns. Access to services is based on citizenship as well as on occupational status.The system is financed by the state budget, social insurance contributions and private payments.The largest share of health expenditure constitutes private expenditure, mainly in the form of out of pocket payments which is also the element contributing most to the overall increase in health expenditure. The delivery of health care services is based on both public and private providers. The presence of private providers is more obvious in primary care,especially in diagnostic technologies, private physicians' practices and pharmaceuticals. Despite success in improving the health of the population, the Greek health care system faces serious structural problems concerning the organization, financing and delivery of services. It suffers from the absence of cost-containment measures and defined criteria for funding, resulting in sickness funds experiencing economic constraints and budget deficits. The high percentage of private expenditure goes against the principle of fair financing and equity in access to health care services. Efficiency is in question due to the lack of incentives to improve performance in the public sector. Mechanisms for needs assessment and priority-setting are underdeveloped and, as a consequence, the regional distribution of health resources is unequal. Centralization of the system is coupled with a lack of planning and coordination, and limited managerial and administrative capacity. In addition, the oversupply of physicians, the absence of a referral system, and irrational pricing and reimbursement policies are factors encouraging under-the-table payments and the black economy. These shortcomings result in low satisfaction with the health care system expressed by citizens. The landmark in the development of the Greek health care system was the creation of the national health system (ESY) in 1983. This report describes the development of the ESY at the structural level and generally, the process of implementing reforms. The strategic targets of health reform initiatives have been to structure a unified health care sector along the lines of the original ESY proposal and to cope with current inefficiencies. However, the three reforms attempted in the 1990s were never fully implemented, while the ambitious reform project of the period 2000–2004, which provided for the regionalization of the system, new management structures, prospective reimbursement, new employment conditions for hospital doctors, modernization of public health services and reorganization of primary health care, was abolished after the elections of 2004 and a change in government. While the new strategy, launched in 2005 with the stated aims of securing the financial viability of the health care system in the short term and its sustainability in the long term, addressed specific weaknesses, it has been rather controversial: the introduction of a centralized administrative public procurement system, the development of public–private partnerships (PPPs) for the construction of public hospitals and the reform of pharmaceutical care have been accompanied by the abolition of professional hospital management and its replacement by political administration. The dominance of clientelism and party thinking instead of consensus-building has resulted in a health policy that lacks continuity and the ability to bring about change.


Mice lacking angiotensin-converting enzyme have low blood pressure, renal pathology, and reduced male fertility.
Mammals produce two isozymes of angiotensin-converting enzyme (ACE). Somatic ACE plays an important role in the control of blood pressure. The function of testis ACE, produced by male and germ cells, is not known. To examine the roles of these isozymes, we used targeted homologous recombination to introduce a modified ACE allele into a mouse line. Mice homozygous for this mutant allele lack both ACE isozymes and have markedly reduced blood pressures. Contrary to a previous report, we found heterozygous male mice to have normal blood pressures. Homozygous mutant mice also have severe renal disease. The renal papilla is markedly reduced, and the intrarenal arteries exhibit vascular hyperplasia associated with a perivascular inflammatory infiltrate. These animals cannot effectively concentrate urine. They also have an abnormally low urinary sodium to potassium ratio despite reduced levels of aldosterone. Homozygous mutant male mice sire significantly smaller litters than wild-type male mice; however, no defect in sperm number, morphology, or motility was detected. ACE-deficient animals demonstrate the role of this enzyme in systemic blood pressure, renal development and function, and male fertility.


Long-term storage limits PCR-based analyses of malaria parasites in archival dried blood spots.
Blood samples collected in epidemiological and clinical investigations and then stored, often at room temperature, as blood spots dried on a filter paper have become one of the most popular source of material for further molecular analyses of malaria parasites. The dried blood spots are often archived so that they can be used for further retrospective investigations of parasite prevalence, or as new genetic markers come to the fore. However, the suitability of the template obtained from dried blood spots that have been stored for long periods for DNA amplification is not known. DNA from 267 archived blood spots collected over a period of 12 years from persons with microscopically confirmed Plasmodium falciparum infection was purified by one of two methods, Chelex and Qiagen columns. These templates were subjected to highly sensitive nested PCR amplification targeting three parasite loci that differ in length and/or copy number. When a 1.6 kb fragment of the parasites' small subunit ribosomal RNA was targeted (primary amplification), the efficiency of P. falciparum detection decreased in samples archived for more than six years, reaching very low levels for those stored for more than 10 years. Positive amplification was generally obtained more often with Qiagen-extracted templates. P. falciparum could be detected in 32 of the 40 negative Qiagen-extracted templates when a microsatellite of about 180 bp was targeted. The remaining eight samples gave a positive amplification when a small region of 238 bp of the higher copy number (20 to 200) mitochondrial genome was targeted. The average length of DNA fragments that can be recovered from dried blood spots decreases with storage time. Recovery of the DNA is somewhat improved, especially in older samples, by the use of a commercial DNA purification column, but targets larger than 1.5 kb are unlikely to be present 10 years after the initial blood collection, when the average length of the DNA fragments present is likely to be around a few hundred bp. In conclusion, the utility of archived dried blood spots for molecular analyses decreases with storage time.


Using a general theory of time and change in patient monitoring: experiment and evaluation.
In this paper, we propose to use one of the well-known general theories of time and change, namely the Event Calculus (Kowalski and Sergot, New Generation Computing 4, 67-95, 1986), to represent temporal aspects in intelligent medical monitoring systems. In particular, we explore the application of CEC (Chittaro and Montanari, Computational Intelligence 12, 359-382, 1996) (an efficient implementation of the Event Calculus) to the management of mechanical ventilation. First, we present the prototype we have built, which has been extensively tested on patient's data from real clinical cases. Then, we provide a thorough evaluation of the obtained results, pointing out both strengths and weaknesses of the approach, and identifying a number of extensions which can be extremely useful to scale up the medical application of the approach.


Initiation of gastrulation in the mouse embryo is preceded by an apparent shift in the orientation of the anterior-posterior axis.
It is generally assumed that the migration of anterior visceral endoderm (AVE) cells from a distal to a proximal position at embryonic day (E)5.5 breaks the radial symmetry of the mouse embryo, marks anterior, and conditions the formation of the primitive streak on the opposite side at E6.5. Transverse sections of a gastrulating mouse embryo fit within the outline of an ellipse, with the primitive streak positioned at one end of its long axis. How the establishment of anterior-posterior (AP) polarity relates to the morphology of the postimplantation embryo is, however, unclear. Transverse sections of prestreak E6.0 embryos also reveal an elliptical outline, but the AP axis, defined by molecular markers, tends to be perpendicular to the long axis of the ellipse. Subsequently, the relative orientations of the AP axis and of the long axis change so that when gastrulation begins, they are closer to being parallel, albeit not exactly aligned. As a result, most embryos briefly lose their bilateral symmetry when the primitive streak starts forming in the epiblast. The change in the orientation of the AP axis is only apparent and results from a dramatic remodeling of the whole epiblast, in which cell migrations take no part. These results reveal a level of regulation and plasticity so far unsuspected in the mouse gastrula.


Protonation equilibria and pore-opening structure of the dual-histidine influenza B virus M2 transmembrane proton channel from solid-state NMR.
The influenza A and B viruses are the primary cause of seasonal flu epidemics. Common to both viruses is the M2 protein, a homotetrameric transmembrane proton channel that acidifies the virion after endocytosis. Although influenza A M2 (AM2) and B M2 (BM2) are functional analogs, they have little sequence homology, except for a conserved HXXXW motif, which is responsible for proton selectivity and channel gating. Importantly, BM2 contains a second titratable histidine, His-27, in the tetrameric transmembrane domain that forms a reverse WXXXH motif with the gating tryptophan. To understand how His-27 affects the proton conduction property of BM2, we have used solid-state NMR to characterize the pH-dependent structure and dynamics of His-27. In cholesterol-containing lipid membranes mimicking the virus envelope, 15N NMR spectra show that the His-27 tetrad protonates with higher pKa values than His-19, indicating that the solvent-accessible His-27 facilitates proton conduction of the channel by increasing the proton dissociation rates of His-19. AM2 is inhibited by the amantadine class of antiviral drugs, whereas BM2 has no known inhibitors. We measured the N-terminal interhelical separation of the BM2 channel using fluorinated Phe-5. The interhelical 19F-19F distances show a bimodal distribution of a short distance of 7 Å and a long distance of 15-20 Å, indicating that the phenylene rings do not block small-molecule entry into the channel pore. These results give insights into the lack of amantadine inhibition of BM2 and reveal structural diversities in this family of viral proton channels.


Analysis of dibenzo[def,p]chrysene-deoxyadenosine adducts in wild-type and cytochrome P450 1b1 knockout mice using stable-isotope dilution UHPLC-MS/MS.
The polycyclic aromatic hydrocarbon (PAH), dibenzo[def,p]chrysene (DBC; also known as dibenzo[a,l]pyrene), is a potent carcinogen in animal models and a class 2A human carcinogen. Recent investigations into DBC-mediated toxicity identified DBC as a potent immunosuppressive agent similar to the well-studied immunotoxicant 7,12-dimethylbenz[a]anthracene (DMBA). DBC, like DMBA, is bioactivated by cytochrome P450 (CYP) 1B1 and forms the reactive metabolite DBC-11,12-diol-13,14-epoxide (DBCDE). DBCDE is largely responsible for the genotoxicity associated with DBC exposure. The immunosuppressive properties of several PAHs are also linked to genotoxic mechanisms. Therefore, this study was designed to identify DBCDE-DNA adduct formation in the spleen and thymus of wild-type and cytochrome P450 1b1 (Cyp1b1) knockout (KO) mice using a highly sensitive stable-isotope dilution UHPLC-MS/MS method. Stable-isotope dilution UHPLC-MS/MS identified the major DBC adducts (±)-anti-cis-DBCDE-dA and (±)-anti-trans-DBCDE-dA in the lung, liver, and spleen of both WT and Cyp1b1 KO mice. However, adduct formation in the thymus was below the level of quantitation for our method. Additionally, adduct formation in Cyp1b1 KO mice was significantly reduced compared to wild-type (WT) mice receiving DBC via oral gavage. In conclusion, the current study identifies for the first time DBCDE-dA adducts in the spleen of mice supporting the link between genotoxicity and immunosuppression, in addition to supporting previous studies identifying Cyp1b1 as the primary CYP involved in DBC bioactivation to DBCDE. The high levels of DBC-DNA adducts identified in the spleen, along with the known high levels of Cyp1b1 expression in this organ, supports further investigation into DBC-mediated immunotoxicity.


Rapid de novo assembly of the European eel genome from nanopore sequencing reads.
We have sequenced the genome of the endangered European eel using the MinION by Oxford Nanopore, and assembled these data using a novel algorithm specifically designed for large eukaryotic genomes. For this 860 Mbp genome, the entire computational process takes two days on a single CPU. The resulting genome assembly significantly improves on a previous draft based on short reads only, both in terms of contiguity (N50 1.2 Mbp) and structural quality. This combination of affordable nanopore sequencing and light weight assembly promises to make high-quality genomic resources accessible for many non-model plants and animals.


Time Trends in Atrial Fibrillation-Associated Stroke and Premorbid Anticoagulation: Population-Based Study and Systematic Review.
Background and Purpose- Prevalence of atrial fibrillation (AF) is increasing, but the impact on overall burden of stroke is uncertain, as is the proportion that could be attributed to under anticoagulation. We did a population-based study of AF-associated stroke and a systematic review of time trends in other stroke incidence studies and of rates of premorbid anticoagulation. Methods- The proportion of incident strokes with associated AF was determined in the OXVASC (Oxford Vascular Study; 2002-2017) and in other prospective, population-based stroke incidence studies published before December 2017. Proportions were pooled by Mantel Haenszel methods, and the pooled percentage of cases with premorbid anticoagulation was determined. Analyses were stratified by the age of study population, mid-study year, country, and ethnicity. Results- Of 1928 patients with incident ischemic stroke in OXVASC, 629 (32.6%; 95% CI, 30.5-34.7) were AF associated, consistent with the pooled estimate from 4 smaller studies over the same study period (608/1948; 31.2%, 30.0-32.4; Phet=0.80). The pooled estimate from all studies reporting premorbid AF over 25 million person-years of observation (1960 onwards; 33 reports) was lower (18.6%, 16.8-20.3) and more heterogeneous (Phet<0.0001), but 62% of heterogeneity was explained by the age of study population, study period, country, and ethnicity. The proportion of incident strokes on premorbid anticoagulation increased over time, both for ischemic stroke in OXVASC (2002-2007: 15.1%, 2008-2012: 19.6%, and 2013-2017: 35.9%; Ptrend<0.0001), and across all studies (P=0.002), but the pooled estimates suggested substantial undertreatment even in the most recent periods (2001-2015: 25.7%, 21.1-30.3 and ≥2010: 31.6%, 18.2-44.9). Conclusions- About 1 in 3 incident ischemic strokes are still AF associated, due partly to low rates of anticoagulation for known prior AF, which therefore represents a major public health opportunity to reduce the burden of stroke.


Lack of transmission of Hematodinium sp. in the blue crab Callinectes sapidus through cannibalism.
Hematodinium spp. are parasitic dinoflagellates of marine crustaceans. Outbreaks of Hematodinium sp. have impacted commercial landings of the blue crab Callinectes sapidus in the coastal bays of Virginia and Maryland (USA), with seasonal peaks in prevalence reaching 85%. The life cycle and transmission routes of the parasite in blue crabs are poorly understood. Cannibalism and waterborne transmission may be routes of transmission, although little conclusive evidence has been reported for these modes. We examined cannibalism as a route by a series of experiments wherein we repeatedly fed adult and juvenile crabs the tissues of crabs infected with Hematodinium. In each experiment, feeding was done 3 times over the course of 1 wk. Only 2 of 120 crabs were infected within 7 to 9 d after feeding, and these 2 were likely infected prior to the experimental exposures. Crabs inoculated with hemolymph from infected donors served as positive controls. They developed infections over 11 to 21 d, indicating that the Hematodinium sp. used in the cannibalism trials was infectious at the time of inoculation. Because amphipods also harbor Hematodinium-like infections, we fed tissues of infected crabs to the estuarine amphipod Leptocheirus plumulosus. Hematodinium DNA was detected in amphipods shortly after feeding, but not in animals held for longer periods, nor was it observed in histological preparations. Amphipods did not obtain infections by scavenging infected crab tissues. Our results show that Hematodinium sp. is not effectively transmitted through ingestion of diseased tissues, indicating that cannibalism may not be a major route of transmission for Hematodinium sp. in blue crabs.


Examining the responsiveness of the National Health Insurance Fund to people living with hypertension and diabetes in Kenya: a qualitative study.
To assess the responsiveness of the National Health Insurance Fund (NHIF) Supa Cover benefit package to the needs of individuals with diabetes and hypertension in Kenya. We carried out a qualitative study and collected data using key informant interviews (n=39) and focus group discussions (n=4) in two purposively selected counties in Western Kenya. Study participants were drawn from NHIF officials, county government officials, health facility managers, healthcare workers and individuals with hypertension and diabetes who were enrolled in NHIF. We analysed data using a thematic approach. Study participants reported that the NHIF Supa Cover benefit package expanded access to services for people living with hypertension and diabetes. However, the NHIF members and healthcare workers had inadequate awareness of the NHIF service entitlements. The NHIF benefit package inadequately covered the range of services needed by people living with hypertension and diabetes and the benefits package did not prioritise preventive and promotive services. Sometimes patients were discriminated against by healthcare providers who preferred cash-paying patients, and some NHIF-empanelled health facilities had inadequate structural inputs essential for quality of care. Study participants felt that the NHIF premium for the general scheme was unaffordable, and NHIF members faced additional out-of-pocket costs because of additional payments for services not available or covered. Whereas NHIF has reduced financial barriers for hypertension and diabetes patients, to enhance its responsiveness to patient needs, NHIF should implement mechanisms to increase benefit package awareness among members and providers. In addition, preventive and promotive services should be included in NHIF's benefits package and mechanisms to monitor and hold contracted providers accountable should be strengthened.


Dynamics of capillary flow of blood into a microfluidic channel.
In this paper, a novel mathematical approach is devised to analyze the flow of blood from a droplet into a microcapillary channel. Special attention is devoted to estimate the effects of variable hydraulic resistance over different flow regimes, influence of suspended RBC particulates on the non-Newtonian flow characteristics and implications of a dynamically-evolving contact angle. Flow characteristics depicting advancement of the fluid within the microfluidic channel turn out to be typically non-linear, as per relative instantaneous strengths of the capillary forces and viscous resistances. It is found that the greater the 'pseudoplasticity' of the blood, the weaker the retarding shear forces. The driving forces, on the other hand, become stronger with time, on account of a reduction of contact angle with a decrease of blood flow velocity, although this strengthening is less prominent for blood samples with greater 'pseudoplasticity'. It is revealed that RBCs suspended in blood samples have a strong influence on the effective blood viscosity, and consequently, may drive the fluid significantly faster into the microchannel, especially when the characteristic length scales of the suspensions approach the hydraulic radius of the channel.


Deconstructing export of malaria proteins.
The virulence of the malaria parasite Plasmodium falciparum is mediated by parasite proteins exported to the surface of infected erythrocytes. In this issue, Maier et al. (2008) report a screen of malaria parasite genes predicted to be involved in parasite protein export and trafficking within the host erythrocyte and discover that many more than expected are essential for parasite survival in vitro.