Exploratory Radiomic Analysis of Conventional vs. Quantitative Brain MRI: Toward Automatic Diagnosis of Early Multiple Sclerosis.
Conventional magnetic resonance imaging (cMRI) is poorly sensitive to pathological changes related to multiple sclerosis (MS) in normal-appearing white matter (NAWM) and gray matter (GM), with the added difficulty of not being very reproducible. Quantitative MRI (qMRI), on the other hand, attempts to represent the physical properties of tissues, making it an ideal candidate for quantitative medical image analysis or radiomics. We therefore hypothesized that qMRI-based radiomic features have added diagnostic value in MS compared to cMRI. This study investigated the ability of cMRI (T1w) and qMRI features extracted from white matter (WM), NAWM, and GM to distinguish between MS patients (MSP) and healthy control subjects (HCS). We developed exploratory radiomic classification models on a dataset comprising 36 MSP and 36 HCS recruited in CHU Liege, Belgium, acquired with cMRI and qMRI. For each image type and region of interest, qMRI radiomic models for MS diagnosis were developed on a training subset and validated on a testing subset. Radiomic models based on cMRI were developed on the entire training dataset and externally validated on open-source datasets with 167 HCS and 10 MSP. Ranked by region of interest, the best diagnostic performance was achieved in the whole WM. Here the model based on magnetization transfer imaging (a type of qMRI) features yielded a median area under the receiver operating characteristic curve (AUC) of 1.00 in the testing sub-cohort. Ranked by image type, the best performance was achieved by the magnetization transfer models, with median AUCs of 0.79 (0.69-0.90, 90% CI) in NAWM and 0.81 (0.71-0.90) in GM. The external validation of the T1w models yielded an AUC of 0.78 (0.47-1.00) in the whole WM, demonstrating a large 95% CI and a low sensitivity of 0.30 (0.10-0.70). This exploratory study indicates that qMRI radiomics could provide efficient diagnostic information using NAWM and GM analysis in MSP. T1w radiomics could be useful for a fast and automated check of conventional MRI for WM abnormalities once acquisition and reconstruction heterogeneities have been overcome. Further prospective validation is needed, involving more data for better interpretation and generalization of the results.


Reflex myoclonic epilepsy in infancy: a critical review.
Benign myoclonic epilepsy in infancy, classified among the generalised idiopathic epilepsies, is characterised by the occurrence of myoclonic seizures in the first three years of life in otherwise normal infants. Some authors have described cases of myoclonic seizures as a reflex response to sudden unexpected tactile or acoustic stimuli and this clinical entity has been proposed as a separate nosographic syndrome, referred to as "reflex myoclonic epilepsy in infancy" (RMEI). We reviewed all published articles and case reports on RMEI in order to clarify clinical and electroencephalographic findings, with particular attention to outcome and treatment. RMEI appears to be a benign variant of idiopathic myoclonic epilepsy in infancy with specific features that occur in neurologically and developmentally normal children. This rare clinical entity is often under-described and under-diagnosed, and for this reason should be brought to the attention of paediatricians in order to avoid extensive investigations and reassure parents of the lack of long-term complications.


In vitro studies on the impact of human serum on the antibacterial effect of faropenem.
The interaction between faropenem and serum in determining antibacterial effect was studied using three target pathogens and two different in vitro methodologies. Strains of Staphylococcus aureus, Streptococcus pneumoniae, capsulate and non capsulate Haemophilus influenzae, all faropenem MIC 0.12 mg/L, were tested in a range of pharmacologically realistic faropenem concentrations with various proportions of serum up to 75%. Using simulated serum bactericidal titres the presence of human serum reduced the activity of faropenem against S. pneumoniae as did heat-treated serum for non-capsulate H. influenzae. Serum on its own was bactericidal against H. influenzae probably due to the presence of complement. The antibacterial effects of combinations of faropenem and serum was assessed in time-kill curves by calculation of the area-under-the bacterial-kill-curve (AUBKC). This was then related to faropenem concentration and the proportion of serum using three-dimensional plots. Serum on its own was mildly inhibitory of the growth of S. aureus, supported improved growth of S. pneumoniae at some proportions and was rapidly bactericidal to H. influenzae, especially the non-capsulate strains. Faropenem had a marked antibacterial effect against all three species in the range 0-2.5 mg/L. Increasing the faropenem concentration from 2.5-10 mg/L produced little or no additional effect. The combination of serum and faropenem had little impact on the antibacterial effect against S. pneumoniae and S. aureus but free drug concentrations were likely to be greater than the MIC in all the combinations used. Against capsulate H. influenzae the effect of serum and faropenem was broadly equivalent while against non-capsulate strains the activity of serum was so great it is difficult to assess the impact of faropenem alone. The interaction between serum and antibiotic in determining antibacterial effect is complex and critically dependent on the proportions of serum and drug concentrations chosen. Three-dimensional plots offer a tool to visualise these complex relationships which may be species specific.


The changes in systemic monocytes in humans undergoing surgical decompression for degenerative cervical myelopathy may influence clinical neurological recovery.
Degenerative cervical myelopathy (DCM) is the most common cause of non-traumatic spinal cord injury worldwide. Surgical decompression is recommended as the preferred treatment strategy for DCM as it halts disease progression and improves neurologic symptoms. We previously demonstrated that neuroinflammation, including monocytes, plays a critical role in the pathobiology of DCM and in ischemic-reperfusion injury (IRI) following surgical decompression. Monocytes are able to enter the spinal cord and brain tissues due to damage to the blood spinal cord and blood brain barrier following injury. Studies have demonstrated that stroke patients and individuals undergoing hip replacement surgery have increased systemic levels of monocytes. Additionally, changes in the signalling responses of monocytes are associated with post-surgical recovery or with ischemic neural tissue damage. Herein, we investigated the role of systemic monocytes as a predictive biomarker for clinical recovery following decompressive surgery for DCM. There was a 2-fold increase in the number of monocytes in DCM patients at 24 h following decompression as compared to baseline levels, which was associated with a significant improvement in the modified Japanese Orthopedic Association scale (mJOA) at 6-months after surgery (p < .0001). In a mouse model of DCM, depleting acute monocytes reduced the non-classical (Ly6Clow) subset from circulation (p < .05) and resulted in a 1.8-fold increase in CD11b expression in the spinal cord at 5 weeks following decompression. Acute monocyte depletion was accompanied by a modest decline in long-term overground locomotion, as evidenced by significantly reduced hindlimb swing speed. This work demonstrated that decompressive surgery leads to an acute increase in peripheral monocytes in human DCM patients, which is modestly associated with clinical recovery. We anticipate that this work could contribute to the implementation of routine measurements of blood monocyte subsets, their activation state, and production of cytokines following decompressive surgery. This information could help to select perioperative anti-inflammatory treatments that can enhance the beneficial effects of decompressive surgery and reduce the incidence of post-operative complications, while avoiding a reduction in systemic monocytes.


Role of the non-neuronal human cholinergic system in lung cancer and mesothelioma: possibility of new therapeutic strategies.
Acetylcholine (Ach), one of the most important examples of a neurotransmitter, represents a phylogenetically old molecule, widely distributed from bacteria to humans. The finding that neuronal Ach receptors (nAChRs) are present in non-neuronal cells raised some interesting issues related to their specific activity. In humans, different studies have showed that many lung cancer cells expressed nAchRs and that low concentrations of nicotine blocked the induction of apoptosis in these cells. A recent study presents data that SCLC express a cholinergic autocrine loop that can regulate cell growth. Such work demonstrates that SCLC cells have a cholinergic phenotype and that ACh exerts as an autocrine growth factor in human lung tumors. Recently it has been shown that human malignant pleural mesothelioma express a cholinergic system, involved in cell growth regulation. Hence, mesothelioma cell growth as well as normal mesothelial cells growth is modulated by the cholinergic system in which agonists (i.e. nicotine) has a proliferative effect and antagonists (i.e. curare) has an inhibitory effect. Furthermore apoptosis mechanisms in mesothelioma cells are under the control of the cholinergic system (nicotine antiapoptotic via induction of NF-kappaB complexes and phosphorilation of Bad at Serine(112), curare proapoptotic via G(0)-G(1) arrest p21(waf-1)-dependent, but p53-independent). The involvement of the non-neuronal cholinergic system in lung cancer and mesothelioma appears reasonable and open up new therapeutic strategies.


Impact of immunophenotyping on management of acute leukemias.
The diagnosis of acute leukemias (AL) requires a multiparametric approach in order to apply risk-adapted therapeutic protocols and appreciate the potential outcome of any given patient. Blast cells immunophenotyping is a key test in this issue, yet the information provided by immunophenotyping has become staggering, and it may be difficult to identify relevant characteristics clearly. This manuscript provides a critical review of the literature regarding the importance of immunophenotyping in acute leukemia diagnosis and management. DATA SOURCES AND METHODS The information given here is based on the experience of the authors, on their literature files and on additional material retrieved through articles and reviews covered by the Institute for Scientific Information (ISI) and the Medline database. Studies with proper definition of the patients and sufficient information regarding follow-up were considered. Immunophenotyping allows an early confirmation of AL diagnosis and establishes lineage assignment. Adequate and comprehensive panels of monoclonal antibodies also allow detection of aberrant immunophenotypic profiles of prognostic value or of use in detecting minimal residual disease. A number of unusual immunophenotypic features are also associated with prognosis. The development of new antibodies, new insights in the functional properties of differentiation antigens, and the quantimetric approach of immunophenotyping will keep this field changing. Moreover, as therapeutic protocols evolve, some earlier results need to be reconsidered. Immunophenotyping, together with cytologic, karyotypic and molecular approaches, retains a crucial place in the diagnosis and management of acute leukemias. It remains a rather specialized approach and should be interpreted in a multidisciplinary perspective, considering for each patient the idiosyncrasies possibly relevant to prognosis.


Perinatal outcome of twin pregnancies in women of advanced age.
The aim of this study was to assess the outcome of twin pregnancies in women of advanced age (>or=35 years) compared with women aged 25-29 years old. This population-based retrospective study compared perinatal outcome of twin pregnancies in primiparae aged 35 or older (N = 240) to that of twin pregnancies in primiparae aged 25-29 years (N = 940). Observed outcomes are adjusted for intermediate (mode of conception and hypertension during pregnancy) and confounding variables (level of education). The possible effect of zygosity and chorionicity was tested in a subset of this database, recorded in the East Flanders Prospective Twin Survey (EFPTS). In twin pregnancies, maternal age of 35 or over is associated with a lower incidence of preterm birth [adjusted odds ratio (AOR) 0.59, 95% confidence interval (CI) 0.44-0.79] and low birthweight (AOR 0.75, 95% CI 0.58-0.98) compared with younger women. Differences in zygosity and chorionicity between both cohorts do not seem to affect the result. In comparison with primiparae aged 25-29 years, perinatal outcome of twin pregnancies is more favourable in primiparae aged 35 or over.


Chitooligosaccharides prevent osteopenia by promoting bone formation and suppressing bone resorption in ovariectomised rats: possible involvement of COX-2.
Chitooligosaccharides (CHOS) added in diet have been found as potent calcium fortifiers in conditions of Ca(2+) deficiency such as osteoporosis. In this study, we found that pharmaceutical intervention using CHOS prevented ovariectomy (OVX)-induced bone mineral density loss and the deterioration of trabecular microarchitecture in a dose-dependent manner (p < 0.05 or 0.01). CHOS (125, 250 mg/kg) suppressed the serum levels of bone resorption biomarkers CTx and TRACP5b induced by OVX (p < 0.05), but increased the levels of osteogenic markers ALP and OC by 11.3-11.6% and 10.7-15.2% of OVX group (p < 0.05), suggesting the exact pharmacological action of CHOS in the control of osteoporosis which may be the result of both promoting bone formation and suppressing bone resorption. Bone turnover-modulating effects of CHOS appear related to their anti-inflammatory capacity to down-regulate mRNA and protein expression of COX-2 (17.2-32.2% and 16.4-21.9% of OVX group, p < 0.05 or 0.01), a key mediator linking between inflammation and osteoporosis.


Glucocorticoids and microbiota regulate ontogeny of intestinal fucosyltransferase 2 requisite for gut homeostasis.
At weaning, the intestinal mucosa surface glycans change from predominantly sialylated to fucosylated. Intestinal adaptation from milk to solid food is regulated by intrinsic and extrinsic factors. The contribution by glucocorticoid, an intrinsic factor, and colonization by microbiota, an extrinsic factor, was measured as the induction of α1,2/3-fucosyltransferase and sucrase-isomaltase (SI) activity and gene expression in conventionally raised, germ-free, and bacteria-depleted mice. In conventionally raised mice, cortisone acetate (CA) precociously accelerated SI gene expression up to 3 weeks and fut2 to 4 weeks of age. In germ-free mice, CA treatment induces only SI expression but not fucosyltransferase. In post-weaning bacteria-deficient (germ-free and bacteria-depleted) mice, fut2 expression remains at low suckling levels. In microbiota deficient mice, intestinal fut2 (but not fut1, fut4 or fut7) was induced only by adult microbiota, but not immature microbiota or CA. Fut2 induction could also be restored by colonization by Bacteroides fragilis, but not by a B. fragilis mutant unable to utilize fucose. Restoration of fut2 expression (by either microbiota or B. fragilis) in bacteria-depleted mice is necessary for recovery from dextran sulfate sodium-induced mucosal injury. Thus, glucocorticoids and microbes regulate distinct aspects of gut ontogeny: CA precociously accelerates SI expression and, only in colonized mice, fut2 early expression. The adult microbiota is required for the fut2 induction responsible for the highly fucosylated adult gut phenotype and is necessary for recovery from intestinal injury. Fut2-dependent recovery from inflammation may explain the high incidence of inflammatory disease (Crohn's and necrotizing enterocolitis) in populations with mutant FUT2 polymorphic alleles.


Clinical course of endomyocardial fibrosis.
The survival pattern, morbidity, and clinical course of 145 patients with endomyocardial fibrosis who were followed up between November 1975 and June 1987 were studied. The diagnosis was confirmed in all cases by cardiac angiography, or echocardiography, or necropsy. Percentage survival at the end of one and 9.5 years was 76.11 and 26.35 respectively. History, physical examination, electrocardiography, and cardiac catheterisation were studied at the first presentation. The determinants of early mortality were studied by univariate Kaplan-Meier estimates compared by the log rank test and Cox proportional hazards multiple regression analysis. Significant univariate predictors of early mortality were QRS axis above +90 degrees, intraventricular conduction delay (QRS duration greater than 0.12 s), duration of symptoms before presentation, New York Heart Association functional classes III and IV, presence of embolic episodes, right atrial mean pressures greater than 20 mm Hg, right ventricular end diastolic pressure greater than 20 mm Hg, and aortic oxygen saturation less than 85%. The significant multivariate predictors of mortality were cyanosis, New York Heart Association functional class at first presentation, and right atrial mean pressure greater than 20 mm Hg. The bleak prognosis of endomyocardial fibrosis did not substantially improve despite advances in the medical management of congestive cardiac failure during the period of the study.


Using the canonical modelling approach to simplify the simulation of function in functional-structural plant models.
Functional-structural plant models that include detailed mechanistic representation of underlying physiological processes can be expensive to construct and the resulting models can also be extremely complicated. On the other hand, purely empirical models are not able to simulate plant adaptability and response to different conditions. In this paper, we present an intermediate approach to modelling plant function that can simulate plant response without requiring detailed knowledge of underlying physiology. Plant function is modelled using a 'canonical' modelling approach, which uses compartment models with flux functions of a standard mathematical form, while plant structure is modelled using L-systems. Two modelling examples are used to demonstrate that canonical modelling can be used in conjunction with L-systems to create functional-structural plant models where function is represented either in an accurate and descriptive way, or in a more mechanistic and explanatory way. We conclude that canonical modelling provides a useful, flexible and relatively simple approach to modelling plant function at an intermediate level of abstraction.


Effects of positive end expiratory pressure during ventilation of the preterm infant.
Twenty two babies receiving artificial ventilator support were studied on 29 occasions to determine the effects of low levels of positive end expiratory pressure. Mean positive end expiratory pressure during these studies was 2.6 cm H2O. Changes in tidal volume, minute volume, compliance, and transcutaneous gas trends produced by the use of positive end expiratory pressure were investigated. Positive end expiratory pressure consistently caused a rise in transcutaneous oxygen tension. Changes in transcutaneous carbon dioxide tension after the introduction of positive end expiratory pressure were less consistent and not of the same magnitude as the observed reduction in minute ventilation. Compliance values fell with the use of positive end expiratory pressure.


A randomized controlled study on the accuracy of free-handed, pilot-drill guided and fully guided implant surgery in partially edentulous patients.
To compare the accuracy of free-handed (FH), pilot-drill guided (PG) and fully guided (FG) implant surgery. Partially edentulous patients in need of ≥2 implants in the posterior maxilla were randomly allocated to one of the following treatment groups: FH, PG and FG. Ideal implant positions were determined in designated software following the fusion of bony information (CBCT data in DICOM format) to the prosthetic wax-up (optical scan data in STL format). The position of every implant as surgically realized was compared to its "ideal position". The apical global deviation (AGD) was the primary outcome of the study. Secondary outcome variables were angular deviation (AD), coronal global deviation (CGD), coronal lateral deviation (CLD), coronal vertical deviation (CVD), apical lateral deviation (ALD) and apical vertical deviation (AVD). Eleven patients (mean age 57; three males; eight females; altogether 26 implants) were treated by FH surgery, 11 (mean age 53; four males; seven females; altogether 24 implants) by PG surgery and 10 (mean age 60; four males; six females; altogether 21 implants) by FG surgery. FG surgery was most accurate (mean AGD: 0.97 mm; maximum AGD: 1.98 mm) followed by PG surgery (mean AGD: 1.43 mm; maximum AGD: 2.72 mm). FH surgery resulted in huge deviation from the ideal position (mean AGD: 2.11 mm; maximum AGD: 4.84 mm). The results on most secondary outcome variables followed the same order. Although screw-retained restorations were planned for all implants, five of 26 in the FH group and one of 24 in the PG group had to be restored by means of a cement-retained restoration. When perfect implant positioning is required, FG surgery should be considered the gold standard approach.


Receptor-like proteins involved in plant disease resistance.
SUMMARY Race-specific resistance in plants against microbial pathogens is governed by several distinct classes of resistance (R) genes. This review focuses on the class that consists of the plasma membrane-bound leucine-rich repeat proteins known as receptor-like proteins (RLPs). The first isolated resistance genes of the RLP class are the tomato Cf genes, which confer resistance to the fungal pathogen Cladosporium fulvum. To date, several other RLP genes are known to be implicated in resistance in other plant-pathogen interactions. These include HcrVf2 from apple, Ve1 and Ve2 from tomato, and RPP27 from Arabidopsis, which are involved in resistance to Venturia, Verticillium and Peronospora, respectively. Furthermore, the tomato RLP gene LeEix initiates defence responses upon elicitation with a fungal ethylene-inducing xylanase (EIX) of non-pathogenic Trichoderma. The tomato Cf genes, which are the most intensively studied RLP resistance genes, are usually found in clusters of several homologues. Whereas some of these homologues are functional Cf resistance genes, others have no known function in resistance. Different evolutionary processes contribute to variation in functional Cf genes, and functional as well as non-functional homologues may provide a source for the generation of novel Cf resistance genes. To date, little is known of the proteins that interact with Cf proteins to initiate defence responses. In contrast to the LeEix protein and the corresponding EIX elicitor, for which a direct interaction was found, no direct interaction between Cf proteins and the corresponding C. fulvum elicitors has been demonstrated. Analogous to the CLAVATA signalling complex, which comprises an RLP, a receptor-like kinase (RLK) and a small proteineous ligand, Cf proteins may form a complex with RLKs and thus initiate signalling upon recognition of the corresponding elicitors. The presence of RLP resistance genes in diverse plant species suggests that these genes play an important role in the extracellular recognition of plant pathogens.


N-H stretching excitations in adenosine-thymidine base pairs in solution: pair geometries, infrared line shapes, and ultrafast vibrational dynamics.
We explore the N-H stretching vibrations of adenosine-thymidine base pairs in chloroform solution with linear and nonlinear infrared spectroscopy. Based on estimates from NMR measurements and ab initio calculations, we conclude that adenosine and thymidine form hydrogen bonded base pairs in Watson-Crick, reverse Watson-Crick, Hoogsteen, and reverse Hoogsteen configurations with similar probability. Steady-state concentration and temperature dependent linear FT-IR studies, including H/D exchange experiments, reveal that these hydrogen-bonded base pairs have complex N-H/N-D stretching spectra with a multitude of spectral components. Nonlinear 2D-IR spectroscopic results, together with IR-pump-IR-probe measurements, as also corroborated by ab initio calculations, reveal that the number of N-H stretching transitions is larger than the total number of N-H stretching modes. This is explained by couplings to other modes, such as an underdamped low-frequency hydrogen-bond mode, and a Fermi resonance with NH(2) bending overtone levels of the adenosine amino-group. Our results demonstrate that modeling based on local N-H stretching vibrations only is not sufficient and call for further refinement of the description of the N-H stretching manifolds of nucleic acid base pairs of adenosine and thymidine, incorporating a multitude of couplings with fingerprint and low-frequency modes.


Modified Rhodopsins From Aureobasidium pullulans Excel With Very High Proton-Transport Rates.
Aureobasidium pullulans is a black fungus that can adapt to various stressful conditions like hypersaline, acidic, and alkaline environments. The genome of A. pullulans exhibits three genes coding for putative opsins ApOps1, ApOps2, and ApOps3. We heterologously expressed these genes in mammalian cells and Xenopus oocytes. Localization in the plasma membrane was greatly improved by introducing additional membrane trafficking signals at the N-terminus and the C-terminus. In patch-clamp and two-electrode-voltage clamp experiments, all three proteins showed proton pump activity with maximal activity in green light. Among them, ApOps2 exhibited the most pronounced proton pump activity with current amplitudes occasionally extending 10 pA/pF at 0 mV. Proton pump activity was further supported in the presence of extracellular weak organic acids. Furthermore, we used site-directed mutagenesis to reshape protein functions and thereby implemented light-gated proton channels. We discuss the difference to other well-known proton pumps and the potential of these rhodopsins for optogenetic applications.


2022 Canadian Cardiovascular Society Guideline for Use of GLP-1 Receptor Agonists and SGLT2 Inhibitors for Cardiorenal Risk Reduction in Adults.
This guideline synthesizes clinical trial data supporting the role of glucagon-like peptide-1 receptor agonists and sodium-glucose co-transporter 2 inhibitors (SGLT2i) for treatment of heart failure (HF), chronic kidney disease, and for optimizing prevention of cardiorenal morbidity and mortality in patients with type 2 diabetes. It is on the basis of a companion systematic review and meta-analysis guided by a focused set of population, intervention, control, and outcomes (PICO) questions that address priority cardiorenal end points. The Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) system and a modified Delphi process were used. We encourage comprehensive assessment of cardiovascular (CV) patients with routine measurement of estimated glomerular filtration rate, urinary albumin-creatinine ratio, glycosylated hemoglobin (A1c), and documentation of left ventricular ejection fraction (LVEF) when evaluating symptoms of HF. For patients with HF, we recommend integration of SGLT2i with other guideline-directed pharmacotherapy for the reduction of hospitalization for HF when LVEF is > 40% and for the reduction of all-cause and CV mortality, hospitalization for HF, and renal protection when LVEF is ≤ 40%. In patients with albuminuric chronic kidney disease, we recommend integration of SGLT2i with other guideline-directed pharmacotherapy to reduce all-cause and CV mortality, nonfatal myocardial infarction, and hospitalization for HF. We provide recommendations and algorithms for the selection of glucagon-like peptide-1 receptor agonists and SGLT2i for patients with type 2 diabetes and either established atherosclerotic CV disease or risk factors for atherosclerotic CV disease to reduce all-cause and CV mortality, nonfatal stroke, and for the prevention of hospitalization for HF and decline in renal function. We offer practical advice for safe use of these diabetes-associated agents with profound cardiorenal benefits.


Use of Near-Infrared Fluorescence Techniques in Minimally Invasive Surgery for Colorectal Liver Metastases.
Colorectal liver metastases (CRLM) afflict a significant proportion of patients with colorectal cancer (CRC), ranging from 25% to 30% of patients throughout the course of the disease. In recent years, there has been a surge of interest in the application of near-infrared fluorescence (NIRF) imaging as an intraoperative imaging technique for liver surgery. The utilisation of NIRF-guided liver surgery, facilitated by the administration of fluorescent dye indocyanine green (ICG), has gained traction in numerous medical institutions worldwide. This innovative approach aims to enhance lesion differentiation and provide valuable guidance for surgical margins. The use of ICG, particularly in minimally invasive surgery, has the potential to improve lesion detection rates, increase the likelihood of achieving R0 resection, and enable anatomically guided resections. However, it is important to acknowledge the limitations of ICG, such as its low specificity. Consequently, there has been a growing demand for the development of tumour-specific fluorescent probes and the advancement of camera systems, which are expected to address these concerns and further refine the accuracy and reliability of intraoperative fluorescence imaging in liver surgery. While NIRF imaging has been extensively studied in patients with CRLM, it is worth noting that a significant proportion of published research has predominantly focused on the detection of hepatocellular carcinoma (HCC). In this study, we present a comprehensive literature review of the existing literature pertaining to intraoperative fluorescence imaging in minimally invasive surgery for CRLM. Moreover, our analysis places specific emphasis on the techniques employed in liver resection using ICG, with a focus on tumour detection in minimal invasive surgery (MIS). Additionally, we delve into recent developments in this field and offer insights into future perspectives for further advancements.


Relationship between behavioral lifestyle and mental health status evaluated using the GHQ-28 and SDS questionnaires in Japanese factory workers.
In order to clarify the relationships between lifestyle behaviors and mental health status in Japanese workers, the 28-item General Health Questionnaire (GHQ-28) and Zung's Self-rating Depression Scale (SDS) were administered to 254 male factory workers; further, eight of their personal health practices were surveyed using the Health Practice Index (HPI). Few studies have used the GHQ-28 and SDS simultaneously; moreover, to date, no study has included the lifestyle factors in these scales. We used these tools for the concurrent assessment of mental health status and classified the subjects into high-risk and low-risk groups. The GHQ-28 and SDS scores correlated negatively with the HPI scores. In the older age group, these scores decreased significantly corresponding to an increase in the total number of favorable lifestyle practices followed. Significant relationships were observed between the GHQ-28 subscales and the HPI. Multiple logistic regression analysis revealed that four of the eight health practices surveyed were significantly related to the mental health status in all subjects. Thus, good health practices appear to be closely associated with good mental health status. However, to establish our results, larger study populations are required for further studies in various occupational fields.


I've seen enough! Prolonged and repeated exposure to disgusting stimuli increases oculomotor avoidance.
Disgust motivates avoidance of stimuli associated with pathogens. Although disgust primarily inhibits oral and epidermal contact, it may also inhibit perceptual contact, particularly given the outsize role of sensory qualities in eliciting disgust. To examine perceptual avoidance of disgust, we presented images of bodily products or spoiled food paired with neutral images for 12-s trials and recorded eye movements (Experiment 1; N = 127). We found that, overall, these disgusting images were not visually avoided compared to neutral images. However, viewing of disgusting images decreased with prolonged (within-trial) and repeated (between-trial) exposure, and these trends were predicted by self-reported disgust to the images. In Experiment 2 (N = 84), we replicated Experiment 1 with a novel set of disgusting images, as well as other unpleasant image categories (suicide, threat) and pleasant images. We found that disgusting stimuli were viewed less than the other unpleasant image categories, and we again found that viewing of disgusting images decreased with prolonged and repeated exposure. Further, we replicated the finding that disgust ratings predicted decreasing viewing of disgusting images, but only for prolonged exposure (within-trial). Unexpectedly, we found that disgust ratings predicted a similar pattern of decreasing viewing for the suicide and threat images as well. These findings suggest that disgust inhibits perceptual contact, but in competition with motivational processes that steer attention toward pathogen threats. We discuss the implications for measuring disgust with eye tracking. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


[How to respond to a dengue epidemic: overview and experience in Puerto Rico].
Dengue, a viral disease transmitted by mosquitoes, is endemic and frequently epidemic in many tropical countries. Because low-incidence periods vary in length, it is difficult to know in advance when an epidemic will occur. Response efforts, despite being logical, have been counter-productive at times. Furthermore, experience has demonstrated that dengue epidemics last a long time, making it important that government control efforts be sustainable while they last. This article describes priority activities requiring attention in order to minimize the impact of dengue epidemics. Such activities, which in many cases can be adapted to combat other types of epidemics as well, are as follows: (1) establishment of an inter-sectoral action committee, (2) formalization of an emergency action plan, (3) epidemiologic surveillance, (4) diagnostic laboratory testing, (5) mosquito control, (6) protection of sources of employment and special populations, (7) patient care, (8) education of medical personnel, (9) research, and (10) transparency before the mass media. The best way to reduce the ravaging effects of dengue epidemics is to anticipate their emergence so that infection can be prevented and steps can be taken to protect the ill. Relying on improvisation to solve all the problems that arise in moments of crisis is inefficient and reckless.


carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase (cad) regulates Notch signaling and vascular development in zebrafish.
The interplay between Notch and Vegf signaling regulates angiogenesis in the embryo. Notch signaling limits the responsiveness of endothelial cells to Vegf to control sprouting. Despite the importance of this regulatory relationship, much remains to be understood about extrinsic factors that modulate the pathway. During a forward genetic screen for novel regulators of lymphangiogenesis, we isolated a mutant with reduced lymphatic vessel development. This mutant also exhibited hyperbranching arteries, reminiscent of Notch pathway mutants. Positional cloning identified a missense mutation in the carbamoyl-phosphate synthetase 2, aspartate transcarbamylase, and dihydroorotase (cad) gene. Cad is essential for UDP biosynthesis, which is necessary for protein glycosylation and de novo biosynthesis of pyrimidine-based nucleotides. Using a transgenic reporter of Notch activity, we demonstrate that Notch signaling is significantly reduced in cad(hu10125) mutants. In this context, genetic epistasis showed that increased endothelial cell responsiveness to Vegfc/Vegfr3 signaling drives excessive artery branching. These findings suggest important posttranslational modifications requiring Cad as an unappreciated mechanism that regulates Notch/Vegf signaling during angiogenesis.


Repurposing From Oncology to Cardiology: Low-Dose 5-Azacytidine Attenuates Pathological Cardiac Remodeling in Response to Pressure Overload Injury.
Recent evidence suggests that transcriptional reprogramming is involved in the pathogenesis of cardiac remodeling (cardiomyocyte hypertrophy and fibrosis) and the development of heart failure. 5-Azacytidine (5aza), an inhibitor of DNA methylation approved for hematological malignancies, has previously demonstrated beneficial effects on cardiac remodeling in hypertension. The aim of our work was to investigate whether pressure overload is associated with alterations in DNA methylation and if intervention with low-dose 5aza can attenuate the associated pathological changes. C57Bl6/J mice underwent surgical constriction of the aortic arch for 8 weeks. Mice began treatment 4 weeks post-surgery with either vehicle or 5aza (5 mg/kg). Cardiac structure and function was examined in vivo using echocardiography followed by post mortem histological assessment of hypertrophy and fibrosis. Global DNA methylation was examined by immunostaining for 5-methylcytosine (5MeC) and assessment of DNA methyltransferase expression. The results highlighted that pressure overload-induced pathological cardiac remodeling is associated with increased DNA methylation (elevated cardiac 5MeC positivity and Dnmt1 expression). Administration of 5aza attenuated pathological remodeling and diastolic dysfunction. These beneficial changes were mirrored by a treatment-related reduction in global 5MeC levels and expression of Dnmt1 and Dnmt3B in the heart. DNA methylation plays an important role in the pathogenesis of pressure overload-induced cardiac remodeling. Therapeutic intervention with 5aza, at a dose 5 times lower than clinically given for oncology treatment, attenuated myocardial hypertrophy and fibrosis. Our work supports the rationale for its potential use in cardiac pathologies associated with aberrant cardiac wound healing.


M phase-specific expression and phosphorylation-dependent ubiquitination of the ClC-2 channel.
Cl(-) channel activities vary during the cell cycle and are thought to play various roles including regulation of cell volume. We have shown previously that ClC-2 channels are directly phosphorylated and functionally regulated by the M phase-specific cyclin-dependent kinase p34(cdc2)/cyclin B. We investigate here to determine whether the expression levels of ClC-2 channel protein vary during the cell cycle. Immunoblot and immunocytochemical analyses of cells cycle-synchronized by serum depletion/replenishment reveal that ClC-2 channel protein is expressed predominantly at M phase in cells with two nuclei and a clear constriction ring, whereas RNA blot analysis shows that ClC-2 mRNA expression does not change during the cell cycle. Ubiquitin assays reveal that the ClC-2 channels are ubiquitinated at M phase, whereas the magnitude of ubiquitination is suppressed by incubation with olomoucine, an inhibitor of p34(cdc2)/cyclin B, and it is almost completely abolished in ClC-2 channels having an S632A mutation, which cannot be phosphorylated by p34(cdc2)/cyclin B, indicating that ubiquitination of ClC-2 channels requires phosphorylation by M phase-specific p34(cdc2)/cyclin B. Regulation at the post-transcriptional level, including phosphorylation-dependent ubiquitination, may contribute to M phase-specific expression of ClC-2 channels. Cell cycle-dependent regulation of expression at the protein level in addition to the regulation of function suggests that the ClC-2 channel plays a physiological role in the cell cycle.


The kidney--the body's playground for drugs: an overview of renal drug handling with selected clinical correlates.
A greater understanding of transport mechanisms contributing to renal drug handling may be useful in predicting drug clearance and drug interactions. Renal clearance is a dynamic process expressed as the sum of the rates of glomerular filtration and tubular secretion minus the rate of tubular reabsorption. Because the transport of drugs is often against a concentration gradient, renal secretion is mostly an active process involving a variety of transporter mechanisms. Discoveries from molecular biology techniques and gene 'knock-out' experiments have identified a variety of renal tubular proteins responsible for the transport of organic cations, organic anions, neutral and cationic hydrophobic compounds, anionic conjugates and specific agents such as prostaglandins. The discovery of a P-glycoprotein (P-gp) transporter at the apical membrane of renal tubular cells is particularly important. By elucidating compounds that act as substrates, inhibitors or inducers of transport proteins, pharmacologists and clinicians may better understand renal drug clearance. This paper provides a brief overview of several identified renal transport proteins including organic anion transporters, organic cation transporters, ATP-dependent transporters (multidrug resistance [P-gp] and multi-drug resistance associated protein), nucleoside transporters (sodium-dependent purine nucleoside transporter and concentrative nucleoside transporter 1) and peptide transporters. A special focus on known P-gp-mediated drug interactions is included to demonstrate the clinical relevance of transporter protein science. At the patient level, this may lead to novel approaches to alter in vivo pharmacokinetics and improve drug safety through a greater understanding of toxic substrate clearance and drug interactions.


Chemical identity of tryptensin with angiotensin.
Tryptensin, a vasopressor substance generated from human plasma protein fraction IV-4 by trypsin, has been isolated and the amino acid composition analysed. The procedures used for the isolation were: (a) adsorption of the formed tryptensin on Dowex 50W (X2; NH4+ form); (b) gel filtration through Sephadex G-25; (c) cation-exchange chromatography on CM-cellulose; (d) anion-exchange chromatography on DEAE-cellulose; (e) re-chromatography on CM-cellulose; (f) gel filtration on Bio-Gel P-2; (g) partition chromatography on high-pressure liquid chromatography. The homogeneity of the isolated tryptensin was confirmed by thin-layer chromatography and thin-layer electrophoresis. The amino acid analysis of the hydrolysate suggested the following proportional composition: Asp, 1; Val, 1; Ile, 1; Tyr, 1; Phe, 1; His, 1; Arg, 1; Pro, 1. This composition is identical with that of human angiotensin.


Characterization of Sec-translocon-dependent extracytoplasmic proteins of Rickettsia typhi.
As obligate intracellular, vector-borne bacteria, rickettsiae must adapt to both mammalian and arthropod host cell environments. Deciphering the molecular mechanisms of the interactions between rickettsiae and their host cells has largely been hindered by the genetic intractability of these organisms; however, research in other gram-negative pathogens has demonstrated that many bacterial determinants of attachment, entry, and pathogenesis are extracytoplasmic proteins. The annotations of several rickettsial genomes indicate the presence of homologs of the Sec translocon, the major route for bacterial protein secretion from the cytoplasm. For Rickettsia typhi, the etiologic agent of murine typhus, homologs of the Sec-translocon-associated proteins LepB, SecA, and LspA have been functionally characterized; therefore, the R. typhi Sec apparatus represents a mechanism for the secretion of rickettsial proteins, including virulence factors, into the extracytoplasmic environment. Our objective was to characterize such Sec-dependent R. typhi proteins in the context of a mammalian host cell infection. By using the web-based programs LipoP, SignalP, and Phobius, a total of 191 R. typhi proteins were predicted to contain signal peptides targeting them to the Sec translocon. Of these putative signal peptides, 102 were tested in an Escherichia coli-based alkaline phosphatase (PhoA) gene fusion system. Eighty-four of these candidates exhibited signal peptide activity in E. coli, and transcriptional analysis indicated that at least 54 of the R. typhi extracytoplasmic proteins undergo active gene expression during infections of HeLa cells. This work highlights a number of interesting proteins possibly involved in rickettsial growth and virulence in mammalian cells.


Suicide and Self-Harm Related Internet Use.
The rise in Internet use adds a new dimension to suicide prevention. We investigated suicide/self-harm (S/Sh)-related Internet use among patients presenting to hospital with self-harm. We asked 1,198 adult and 315 child and adolescent patients presenting to hospital following self-harm in a city in South West England about Internet use associated with their hospital presentation. Associations between Internet use and sociodemographic and clinical characteristics were investigated using multivariable logistic regression models. Focus groups with clinicians explored the acceptability and utility of asking about Internet use. The prevalence of S/Sh-related Internet use was 8.4% (95% CI: 6.8-10.1%) among adult hospital presentations and 26.0% (95% CI = 21.3-31.2%) among children's hospital presentations. In both samples, S/Sh-related Internet use was associated with higher levels of suicidal intent. Mostly, clinicians found it acceptable to ask about Internet use during psychosocial assessments and believed this could inform perceptions of risk and decision-making. It is unclear whether the findings in this study are applicable to the general self-harm patient population because only those who had psychosocial assessments were included. S/Sh-related Internet use is likely to become increasingly relevant as the Internet-native generation matures. Furthermore, Internet use may be a proxy marker for intent.


Embodied cognition: Is activation of the motor cortex essential for understanding action verbs?
In 8 experiments using language processing tasks ranging from lexical decision to sensibility judgment, participants made hand or foot responses after reading hand- or foot-associated words such as action verbs. In general, response time (RT) tended to be faster when the hand- versus foot-associated word was compatible with the limb that was required to respond (e.g., hand response to a hand-associated word) than when it was incompatible (e.g., foot response to a hand-associated word). To see whether this compatibility effect reflects differential hand- versus foot-specific motor activation produced by the words, as suggested by some embodied theories of language understanding, we monitored 2 event-related potential (ERP) measures previously found to be sensitive to the activation of these limbs. As expected, the ERP results replicated previous findings that the monitored ERPs differ for hand versus foot movements. More importantly, the ERPs provided no evidence of any difference for hand- versus foot-associated words. Thus, the results weaken previous claims that the understanding of action verbs requires activation of the motor areas used to carry out the named action. Instead, they support claims that language-related compatibility effects on RT may arise prior to motor processes, which implies that such effects are not decisive evidence for embodied language understanding. (PsycINFO Database Record


[Reaction of pyrophosphorolysis catalyzed by Escherichia coli RNA polymerase].
E. coli RNA polymerase is shown to be capable of catalyzing consecutive DNA-dependent pyrophosphorolysis of RNA in the presence of inorganic pyrophosphate and Mg2+. Active ternary complex of the enzyme with DNA and nascent RNA is needed for the reaction, the mixure of all the components can not carry out pyrophosphorolysis. The reaction was realized in the absence of added nucleoside triphosphates. Nucleoside triphosphates are low molecular mass products of the reaction. The rate of pyrophosphorolysis of the RNA synthesised for the AI promoter of the DNA of wild type T7 phage and delta D III T7 mutant phage was followed as a function of primary structure of the DNA, temperature, ionic strength and inorganic pyrophosphate concentration. The relative rate pyrophosphorolysis for particular nucleotides in different regions of the RNA can differ by several orders of magnitude depending on the primary structure of the RNA that undergoes pyrophosphorolysis. Ternary complex containing partially pyrophosphorilised RNA is active on the RNA synthesis when pyrophosphate is removed and nucleoside triphosphates are added to the reaction mixture. RNA as short as 70-8 nucleotides long can be produced at the conditions used. It seems that efficient dissociation in this region of RNA limits the pyrophosphorolysis to proceed up to the 5' end of RNA. Ternary complex of RNA polymerase with nascent RNA and DNA is shown to undergo site specific dissociation. The specificity of the dissociation is shown to be a function of the primary structure of RNA and the direction of the reaction. Dissociation occurs at different places along RNA sequence when the RNA is synthesised and when it is pyrophosphorilised.


De Novo Mutation Rate Variation and Its Determinants in Chlamydomonas.
De novo mutations are central for evolution, since they provide the raw material for natural selection by regenerating genetic variation. However, studying de novo mutations is challenging and is generally restricted to model species, so we have a limited understanding of the evolution of the mutation rate and spectrum between closely related species. Here, we present a mutation accumulation (MA) experiment to study de novo mutation in the unicellular green alga Chlamydomonas incerta and perform comparative analyses with its closest known relative, Chlamydomonas reinhardtii. Using whole-genome sequencing data, we estimate that the median single nucleotide mutation (SNM) rate in C. incerta is μ = 7.6 × 10-10, and is highly variable between MA lines, ranging from μ = 0.35 × 10-10 to μ = 131.7 × 10-10. The SNM rate is strongly positively correlated with the mutation rate for insertions and deletions between lines (r > 0.97). We infer that the genomic factors associated with variation in the mutation rate are similar to those in C. reinhardtii, allowing for cross-prediction between species. Among these genomic factors, sequence context and complexity are more important than GC content. With the exception of a remarkably high C→T bias, the SNM spectrum differs markedly between the two Chlamydomonas species. Our results suggest that similar genomic and biological characteristics may result in a similar mutation rate in the two species, whereas the SNM spectrum has more freedom to diverge.


Potential Implication of Azole Persistence in the Treatment Failure of Two Haematological Patients Infected with Aspergillus fumigatus.
Invasive aspergillosis (IA) is a major cause of morbidity and mortality in patients receiving allogeneic haematopoieticcell transplantation. The deep immunosuppression and a variety of potential additional complications developed in these patients result in IA reaching mortality rates of around 50-60%. This mortality is even higher when the patients are infected with azole-resistant isolates, demonstrating that, despite the complexity of management, adequate azole treatment can have a beneficial effect. It is therefore paramount to understand the reasons why antifungal treatment of IA infections caused by azole-susceptible isolates is often unsuccessful. In this respect, there are already various factors known to be important for treatment efficacy, for instance the drug concentrations achieved in the blood, which are thus often monitored. We hypothesize that antifungal persistence may be another important factor to consider. In this study we present two case reports of haematological patients who developed proven IA and suffered treatment failure, despite having been infected with susceptible isolates, receiving correct antifungal treatment and reaching therapeutic levels of the azole. Microbiological analysis of the recovered infective isolates showed that the patients were infected with multiple strains, several of which were persisters to voriconazole and/or isavuconazole. Therefore, we propose that azole persistence may have contributed to therapeutic failure in these patients and that this phenomenon should be considered in future studies.


Falls in urban children. A problem revisited.
Falls in urban setting are a common cause for emergency room visits in children and adolescents. In a retrospective review, the charts of 48 patients admitted between 1980 and 1985 with a history of a vertical fall from a height were examined. In comparison, a previous review from the same institution disclosed that 66 patients were admitted because of a vertical fall from a height between 1965 and 1974, suggesting an increase of 37.5%. Most children fell from heights of 12 ft or less, although an increasing proportion of children in our series (33%) fell from heights of 36 ft or less. Sites included windows, walls, and roofs. The peak age of incidence has increased from 2 to 6 years; however, the mean age of children in whom significant injury occurred was 7.5 years, with only 27% of children under 3 years of age suffering a documented injury, as opposed to 67% of children over 3 years of age. Children are more apt to suffer a fracture than any other injury, most likely a fracture of the ulna and/or radius. Although hospital costs are high, mortality rates (2%) and the incidence of long-term sequelae (4%) are low. In conclusion, falls in the urban setting continue to be a significant public health problem, particularly in the 6- to 7-year age group.


Idarubicin, cytarabine, and nivolumab in patients with newly diagnosed acute myeloid leukaemia or high-risk myelodysplastic syndrome: a single-arm, phase 2 study.
Outcomes for younger patients with acute myeloid leukaemia have moderately improved over the past two decades owing to better supportive care and recent introduction of novel targeted agents. Blocking PD-1 and its ligand's pathways enhances antileukaemia responses by enabling T cells in murine models. We aimed to assess the addition of nivolumab to frontline therapy with idarubicin and cytarabine in patients with newly diagnosed acute myeloid leukaemia or high-risk myelodysplastic syndrome. This single-arm, phase 2 part of the phase 1-2 study of nivolumab in combination with idarubicin and cytarabine was done at the University of Texas MD Anderson Cancer Center (Houston, TX, USA). Eligible patients were aged 18-60 years (or >60 years if suitable for intensive chemotherapy), and had newly diagnosed acute myeloid leukaemia or high-risk myelodysplastic syndrome, and an Eastern Cooperative Oncology Group performance status of 0-2. Induction included cytarabine 1·5 g/m2 by 24-h continuous infusion daily on days 1-4 (3 days in patients >60 years) and idarubicin 12 mg/m2 daily on days 1-3. Nivolumab 3 mg/kg was started on day 24 (range 22-26) and continued every 2 weeks for up to a year in responders. Responders received either up to five consolidation cycles of attenuated doses of idarubicin and cytarabine, or allogeneic stem cell transplantation if eligible. The primary endpoint was event-free survival. Efficacy and safety analyses were done in all patients who received at least one dose of study treatment. Secondary endpoints were relapse-free survival and overall survival. This ongoing trial is registered with ClinicalTrials.gov, number NCT02464657. Between Aug 7, 2015, and June 2, 2018, 44 patients were enrolled of whom 22 (50%) had adverse genetic risk by European Leukaemia Network classification. All patients were evaluable for safety and efficacy. At a median follow-up of 17·25 months (IQR 0·50-30·40), median event-free survival was not reached (95% CI 7·93-NR). Median relapse-free survival of responders was 18·54 months (95% CI 8·20-23·22). The median overall survival was 18·54 months (95% CI 10·81-28·81). Six patients had seven grade 3-4 immune-related adverse events with two cases of rash, two of colitis, and one each of transaminitis, pancreatitis, and cholecystitis. 19 (43%) of 44 patients achieved a response and proceeded to allogeneic stem cell transplantation, with grade 3-4 graft-versus-host disease observed in five (26%). No treatment related deaths were attributed to nivolumab. Addition of nivolumab to induction chemotherapy with idarubicin and cytarabine is feasible in patients with newly diagnosed acute myeloid leukaemia or high-risk myelodysplastic syndrome. Post-transplant severe graft-versus-host disease could be improved, and earlier initiation of checkpoint inhibitor therapy is warranted in future studies. The MD Anderson Cancer Center Support Grant CA016672, and the MD Anderson Cancer Center Leukaemia SPORE CA100632 from the National Cancer Institute, Bristol Myers Squibb.


Learning strategies of first year nursing and medical students: a comparative study.
Interprofessional education (IPE), where two or more professions learn with, from, and about each other to improve collaboration and the quality of care, has been proposed as a curriculum strategy to promote mutual understanding between professions, thus helping to prepare health professionals to work in challenging contemporary health systems. Although there is support for IPE initiatives within health professional education, differences in student motivation and learning strategies are likely to contribute to the success of these initiatives. To explore self-regulated learning strategies used by first year medical and nursing students, and to determine if these strategies were different among nursing students who were high achievers. A comparative survey design. Nursing and medical nursing schools in a large university in the western region of Sydney, Australia. Six hundred and sixty-five first year nursing (n=565) and medical (n=100) students in a large university in the western region of Sydney were surveyed to assess motivational and learning strategies using The Motivated Strategies for Learning Questionnaire (MSLQ). Data relating to sociodemographic characteristics and academic performance were also collected. Nursing students were significantly older than medical students (mean age: 24.4 years versus 19.4 years; p<0.001), and there were also more females in the nursing student group (82% versus 56%; p<0.001). Although nursing students had a higher mean score for extrinsic goal orientation compared to medical students (p<0.001), medical students had higher mean scores for the other four learning strategies measured: peer learning (p=0.003), help seeking (p=0.008), critical thinking (p=0.058), and time and study environment management (p<0.001). Similarly, the grade point average (GPA) of medical students at the end of their first year was significantly higher (4.5, S.D. 1.4 versus 3.6, S.D. 1.3; p<0.001) compared to that of nursing students. While interprofessional education is seen to have many benefits for students, this study demonstrates differences in motivational and learning strategies between nursing and medical students that may impact on the success of interprofessional programs.


Complete Genome Sequence of Pseudomonas sp. Strain phDV1, an Isolate Capable of Efficient Degradation of Aromatic Hydrocarbons.
Pseudomonas sp. strain phDV1 is a Gram-negative bacterium capable of degrading aromatic hydrocarbons. Here, we present the complete genome sequence of this strain, which consists of 4,727,682 bp, with a 62.3% G+C content and 4,574 genes. Multiple genes responsible for the degradation of aromatics are present in this strain.


Mapping hemodynamic correlates of seizures using fMRI: A review.
Functional magnetic resonance imaging (fMRI) is able to detect changes in blood oxygenation level associated with neuronal activity throughout the brain. For more than a decade, fMRI alone or in combination with simultaneous EEG recording (EEG-fMRI) has been used to investigate the hemodynamic changes associated with interictal and ictal epileptic discharges. This is the first literature review to focus on the various fMRI acquisition and data analysis methods applied to map epileptic seizure-related hemodynamic changes from the first report of an fMRI scan of a seizure to the present day. Two types of data analysis approaches, based on temporal correlation and data driven, are explained and contrasted. The spatial and temporal relationship between the observed hemodynamic changes using fMRI and other non-invasive and invasive electrophysiological and imaging data is considered. We then describe the role of fMRI in localizing and exploring the networks involved in spontaneous and triggered seizure onset and propagation. We also discuss that fMRI alone and combined with EEG hold great promise in the investigation of seizure-related hemodynamic changes non-invasively in humans. We think that this will lead to significant improvements in our understanding of seizures with important consequences for the treatment of epilepsy.


Comparative phylogeography, genetic differentiation and contrasting reproductive modes in three fungal symbionts of a multipartite bark beetle symbiosis.
Multipartite symbioses are complex symbiotic relationships involving multiple interacting partners. These types of partnerships provide excellent opportunities in which to apply a comparative approach to identify common historical patterns of population differentiation and species-specific life history traits. Using three symbiotic blue-stain fungal species (Ophiostomatacea) associated with outbreaking populations of the mountain pine beetle (Dendroctonus ponderosae Hopkins) in western Canada, we applied phylogenetic, population genetic and demographic approaches to clarify phylogeographic patterns among the three fungal species. Broadly, the three species showed significant population differentiation, forming northern and southern populations, despite dramatic differences in haplotype diversity. Finer structuring and population demographic patterns were less consistent, showing some interspecific incongruence. By contrasting these species simultaneously, we were able to identify differences in recombination rate and ecological traits that can explain the observed patterns of incongruence among the fungal species. By applying a comparative approach to partners of a multipartite symbiosis, we were able to distinguish congruent population structuring and species-specific differences that help us to understand the complexity and evolution of this symbiotic system.


Quality of occupational history assessments in working age adults with newly diagnosed asthma.
Approximately 10 to 15% of new-onset asthma in adults is attributable to occupational exposure. The occupational history is the most important instrument in the diagnosis of occupational asthma (OA). To assess the quality of occupational histories obtained by health-care providers and to measure the prevalence of clinician-diagnosed OA in a population at elevated risk for OA. An academic US Department of Veteran Affairs medical center. One hundred ninety-seven adults (age range, 18 to 55 years) with newly diagnosed asthma who had completed pulmonary function testing (PFT) and a structured respiratory health questionnaire. We conducted a structured retrospective comparison of occupational respiratory health history documented by clinicians with data documented by patients on a structured questionnaire. We analyzed PFT results to assess physiologic impairment. We also conducted a structured examination of the actions taken by health-care providers based on their occupational history assessments. Patient self-reports of respiratory exposures and symptoms were common. A job title was documented by one or more clinicians in 75% of patient medical records. Additional occupational history data were charted much less frequently. A diagnosis of OA was made in only 2% of patients. Clinical action to address OA was documented for only one patient. Clinicians who manage adults with newly diagnosed asthma take incomplete occupational histories. We detected discordance between the occupational exposure histories documented by patients and those charted by clinicians. OA may go unrecognized and possibly undermanaged by clinicians.


Corneal Higher-order Aberrations and Visual Improvement Following Corneal Transplantation in Treating Herpes Simplex Keratitis.
To examine corneal higher-order aberrations (HOAs) and visual improvement following corneal transplantation in treating corneal scar caused by herpes simplex keratitis (HSK). Retrospective consecutive case series. This study included a total of 52 eyes: 18 eyes of normal subjects, and 34 eyes of consecutive patients with corneal scar owing to HSK who underwent penetrating keratoplasty (PKP, 17 eyes) or deep anterior lamellar keratoplasty (DALK, 17 eyes). HOAs of the anterior, posterior surfaces and the total cornea were analyzed by anterior segment optical coherence tomography. The correlations between corneal HOAs and visual improvement were also analyzed. Mean logarithm of the minimal angle of resolution (logMAR) visual acuity significantly improved from 1.40 ± 0.70 to 0.46 ± 0.45 after corneal transplantation (P < .0001). Mean corneal HOAs of the anterior surface significantly decreased after corneal transplantation (PKP: from 1.16 ± 0.59 μm to 0.58 ± 0.35 μm, P = .035, DALK: from 0.94 ± 0.57 μm to 0.37 ± 0.18 μm, P = .004). Visual acuity following corneal transplantation was correlated with the corneal HOAs at 12 months (r = 0.53, P = .01). Visual improvement at 3, 6, and 12 months was positively correlated with preoperative HOAs of the total cornea and posterior surface (all P < .01). Preoperative corneal HOAs were associated with the visual improvement in treating corneal scar caused by HSK. Thus, the preoperative assessment of corneal HOAs, especially of the posterior surface, is important in the decision to perform corneal transplantation in eyes with HSK.


Gene-wide association study between the methylenetetrahydrofolate reductase gene (MTHFR) and schizophrenia in the Japanese population, with an updated meta-analysis on currently available data.
Methylenetetrahydrofolate reductase (MTHFR) is a critical molecule for single-carbon transfer reactions. Recent evidence suggests that polymorphisms of MTHFR are related to neural tube deficits and the pathogenesis of schizophrenia. While several studies have demonstrated associations between the gene encoding the MTHFR (MTHFR) polymorphisms and schizophrenia, these studies lack consistency. Therefore, we conducted a gene-wide association study (patients with schizophrenia = 696, control subjects = 747) and performed imputation analysis. Additionally, we performed meta-analysis on currently available data from 18 studies for two common functional polymorphisms (rs1801131 and rs1801133). There were no significant associations with schizophrenia in the single marker analysis for the seven tagging SNPs of MTHFR. In the haplotypic analysis, a nominally significant association was observed between the haplotypes, which included four SNPs (rs1801133, rs17421511, rs17037396, and rs9651118) and the schizophrenic patients. Additionally, the imputation analysis demonstrated there were several associated markers on the MTHFR chromosomal region. However, confirmatory analyses of three tagging SNPs (rs1801133, rs17037396, and rs9651118) and the top SNP (rs17421511) for the imputation results (patients with schizophrenia = 797, control subjects = 1025) failed to replicate the haplotypic analysis and the imputation results. These findings suggest that MTHFR polymorphisms are unlikely to be related to the development of schizophrenia in the Japanese population. However, since our meta-analysis results demonstrated strong support for association of rs1801133 with schizophrenia, further replication studies based on a gene-wide approach need to be considered.


Male infertility, impaired spermatogenesis, and azoospermia in mice deficient for the pseudophosphatase Sbf1.
Pseudophosphatases display extensive sequence similarities to phosphatases but harbor amino acid alterations in their active-site consensus motifs that render them catalytically inactive. A potential role in substrate trapping or docking has been proposed, but the specific requirements for pseudophosphatases during development and differentiation are unknown. We demonstrate here that Sbf1, a pseudophosphatase of the myotubularin family, is expressed at high levels in seminiferous tubules of the testis, specifically in Sertoli's cells, spermatogonia, and pachytene spermatocytes, but not in postmeiotic round spermatids. Mice that are nullizygous for Sbf1 exhibit male infertility characterized by azoospermia. The onset of the spermatogenic defect occurs in the first wave of spermatogenesis at 17 days after birth during the synchronized progression of pachytene spermatocytes to haploid spermatids. Vacuolation of the Sertoli's cells is the earliest observed phenotype and is followed by reduced formation of spermatids and eventual depletion of the germ cell compartment in older mice. The nullizygous phenotype in conjunction with high-level expression of Sbf1 in premeiotic germ cells and Sertoli's cells is consistent with a crucial role for Sbf1 in transition from diploid to haploid spermatocytes. These studies demonstrate an essential role for a pseudophosphatase and implicate signaling pathways regulated by myotubularin family proteins in spermatogenesis and germ cell differentiation.


A statistical field theory of salt solutions of 'hairy' dielectric particles.
In this paper, we formulate a field-theoretical model of dilute salt solutions of electrically neutral spherical colloid particles. Each colloid particle consists of a 'central' charge that is situated at the center and compensating peripheral charges (grafted to it) that are fixed or fluctuating relative to the central charge. In the framework of the random phase approximation, we obtain a general expression for electrostatic free energy of solution and analyze it for different limiting cases. In the limit of infinite number of peripheral charges, when they can be modelled as a continual charged cloud, we obtain an asymptotic behavior of the electrostatic potential of a point-like test charge in a salt colloid solution at long distances, demonstrating the crossover from its monotonic decrease to damped oscillations with a certain wavelength. We show that the obtained crossover is determined by certain Fisher-Widom line. For the same limiting case, we obtain an analytical expression for the electrostatic free energy of a salt-free solution. In the case of nonzero salt concentration, we obtain analytical relations for the electrostatic free energy in two limiting regimes. Namely, when the ionic concentration is much higher than the colloid concentration and the effective size of charge cloud is much bigger than the screening lengths that are attributed to the salt ions and the central charges of colloid particles. The proposed theory could be useful for theoretical description of the phase behavior of salt solutions of metal-organic complexes and polymeric stars.


Subclinical ventricular dysfunction by Speckle Tracking in adult patients with coarctation of the aorta.
The aim of this study was to investigate the presence of early left ventricular (LV) systolic dysfunction in adult patients with aortic coarctation (AoCo) and systemic arterial hypertension (SAH) compared to systemic hypertensive patients without coarctation and healthy controls by speckle tracking. Sixty-one subjects were studied, who attended consecutively to external consultation. All were submitted to clinical history, resting electrocardiogram, conventional echocardiogram and with Speckle Tracking. 15 patients with AoCo and SAH were in functional class NYHA I, and five in functional class NYHA II. A significant inverse correlation was observed between the global longitudinal deformation and the left ventricular mass index in the AoCo and SAH group and in the hypertensive patients (r = 0.53, p = 0.02; r = 0.52, p < 0.0001, respectively). In patients with AoCo and SAH, global longitudinal deformation and left ventricular mass index are significantly related to subclinical LV dysfunction. These parameters are important and should be taken into account to regulate timely therapeutic behavior.


Memantine, an NMDA antagonist, prevents the development of hyperthermia in an animal model for serotonin syndrome.
Serotonin (5-HT) syndrome is the most serious side effect of antidepressants. Although several drugs have been used for the treatment of 5-HT syndrome, a universal pharmacotherapy has not been established. NMDA receptor antagonists have been reported to have neuroprotective effects. In the present study, the efficacy of NMDA antagonists, including memantine and MK-801, and potent 5-HT (2A) antagonists, including risperidone and ketanserin, was evaluated in a 5-HT syndrome animal model. 5-Hydroxy-l-tryptophan (100 mg/kg) and clorgyline (2 mg/kg) were administered intraperitoneally in rats to induce 5-HT syndrome. The rectal temperature of the rats was measured, and the noradrenaline (NA) and 5-HT levels in the anterior hypothalamus were measured using a microdialysis technique. In the group pretreated with saline, the rectal temperature increased to more than 40 degrees C, and all of the animals died within 90 min of the drug's administration. The NA and 5-HT levels in the anterior hypothalamus increased to about 15- and 1100-fold of the pre-administration levels, respectively. Pretreatment with risperidone (0.5 mg/kg) and ketanserin (5 mg/kg) prevented the development of hyperthermia and the increase in the NA level. Memantine (10 mg/kg) and MK-801 (0.5 mg/kg) also prevented the development of hyperthermia and the increase in the NA level. These results suggest that NMDA antagonists, as well as potent 5-HT (2A) antagonists, may be effective drugs for the treatment of 5-HT syndrome. Since memantine is clinically well tolerated, this drug is a particularly promising therapeutic drug for 5-HT syndrome treatment.


The application of high density microarray for analysis of mitogenic signaling and cell-cycle in the adrenal.
Angiotensin II (AII) binds to specific G-protein coupled receptors and is mitogenic in adrenal, liver epithelial, and vascular smooth muscle cells. The H295R human adrenocortical cell line, which expresses AII receptors predominantly of the AT1 subclass, proliferates in response to treatment with AII. The induction and maintenance of cellular proliferation involves a precisely coordinated induction of a variety of genes. As the human genome sequencing projects near completion a variety of high throughput technologies have been developed in order to create dynamic displays of genomic responses. One high throughput method, the gridded cDNA microarray has been developed in which immobilised DNA samples are hybridized on glass slides for the identification of global genomic responses. For this purpose high precision robotic microarrayers have been developed at AECOM. The cyclin D1 gene, which encodes the regulatory subunit of the cyclin D1-dependent kinase (CD1K) required for phosphorylation of the retinoblastoma protein (pRB), was induced by AII in H295R cells. Abundance of the cyclin D1 gene is rate-limiting in G1 phase progression of the cell-cycle in a variety of cell types. AII induced cyclin D1 promoter activity through a c-Fos and c-Jun binding sequence at -954 bp. Theabundance of c-Fos within this complex was increased by AII treatment. Analysis of AII signaling in adrenal cells by cDNA microarray demonstrated an induction of the human homologue of Xenopus XPMC2 (HXPMC2). The cDNA for XPMC2 was previously shown to rescue mitotic catastrophe in mutant S. Pombe defective in cdc2 kinase function. Further studies are required to determine the requirement for cyclin D1 and XPMC2H in AII-induced cell-cycle progression and cellular proliferation in the adrenal.


Nasopharyngeal colonization by Neisseria lactamica and induction of protective immunity against Neisseria meningitidis.
Natural immunity to Neisseria meningitidis may result from nasopharyngeal carriage of closely related commensals, such as Neisseria lactamica. We enrolled 61 students with no current carriage of Neisseria species and inoculated them intranasally with 10,000 colony-forming units of Neisseria lactamica or sham control. Colonization was monitored in oropharyngeal samples over 6 months. We measured specific mucosal and systemic antibody responses to N. lactamica and serum bactericidal antibody (SBA) and opsonophagocytic antibodies to a panel of N. meningitidis serogroup B strains. We also inoculated an additional cohort following vaccination with N. lactamica outer-membrane vesicles (OMV) produced from the same strain. Twenty-six (63.4%) of 41 inoculated individuals became colonized with N. lactamica; 85% remained colonized at 12 weeks. Noncarriers were resistant to rechallenge, and carriers who terminated carriage were relatively resistant to rechallenge. No carriers acquired N. meningitidis carriage over 24 weeks, compared with 3 control subjects (15%). Carriers developed serum IgG and salivary IgA antibodies to the inoculated N. lactamica strain by 4 weeks; noncarriers and control subjects did not. Cross-reactive serum bactericidal antibody responses to N.meningitidis were negligible in carriers, but they developed broad opsonophagocytic antimeningococcal antibodies. OMV vaccinees developed systemic and mucosal anti-N. lactamica antibodies and were relatively resistant to N. lactamica carriage but not to natural acquisition of N. meningitidis. Carriers of N. lactamica develop mucosal and systemic humoral immunity to N. lactamica together with cross-reacting systemic opsonophagocytic but not bactericidal antibodies to N. meningitidis. Possession of humoral immunity to N. lactamica inhibits acquisition of N. lactamica but not of N. meningitidis. Some individuals are intrinsically resistant to N. lactamica carriage, independent of humoral immunity.


Role of childhood health in the explanation of socioeconomic inequalities in early adult health.
To examine the contribution of childhood health to the explanation of socioeconomic inequalities in health in early adult life. Retrospective data were used, which were obtained from a postal survey in the baseline of a prospective cohort study (the Longitudinal Study on Socio-Economic Health Differences in the Netherlands). Adult socioeconomic status was indicated by educational level, while health was indicated by perceived general health. Childhood health was measured by self reported periods of severe disease in childhood. Relations were analysed using logistic regression models. The reduction in odds ratios of "less than good" perceived general health for different educational groups after adjustment for childhood health was used to estimate the contribution of childhood health. The population of the city of Eindhoven and surroundings in the south east of the Netherlands in 1991. 2511 respondents, aged 25-34 years, men and women, of Dutch nationality, were included in the analysis. There was a clear association between childhood health and adult health, as well as an association between childhood health and adult socioeconomic status. Approximately 5% to 10% of the increased risk of the lower socioeconomic groups of having a "less than good" perceived general health can be explained by childhood health. Childhood health contributes to the explanation of socioeconomic inequalities in early adult health. Although this contribution is not very large, it cannot be ignored and has to be interpreted largely in terms of selection on health.


Mortality outcomes between pregnant women booked for antenatal care and unbooked pregnant women.
Perinatal fetal mortality and maternal mortality remains relatively high in Saudi Arabia. Antenatal care aims to improve outcomes; however, evidence to demonstrate its impact on outcomes in Saudi Arabia is lacking. Investigate whether booking status for antenatal care impacted outcomes for pregnant women and identify outcomes that predicted booking status. Retrospective observational cohort study. Tertiary care center. All Saudi women admitted to the labor or delivery units between January 2011 and December 2019 were included. Outcomes were compared between booked and unbooked women, and logistic regression was used to identify outcomes that predicted booking status, adjusted for age. Booked/unbooked status and perinatal mortality of Saudi women. 10 781 women; 9546 (88.5%) booked, (11.1%) 1192 unbooked. Unbooked mothers had higher incidences of meconium-stained liquor (P=.040), ruptured uterus (P=.017), and blood loss >1000 mL during cesarean deliveries (P=.003), but a lower episiotomy rate (P<.001). Perinatal fetal mortality and maternal mortality were equivalent between the two groups. Episiotomy, higher birth weight, higher mother age, perinatal death, delivery mode, onset of labor, and use of analgesics were all independent predictors of the mother being booked when adjusted for all outcomes and age. The rate of perinatal and maternal mortality in this cohort was relatively low and equivalent between booked and un-booked mothers, contrary to our expectations. More analysis of the socioeconomic data may explain this striking result. Retrospective chart-review with incomplete data retrieval that affected the completeness of data retrieved. The results of the multivariate analysis cannot be used to infer causality because the study is observational.


Citrus Peel Extract Ameliorates High-Fat Diet-Induced NAFLD via Activation of AMPK Signaling.
Non-alcoholic fatty liver disease (NAFLD) is prevalent in the elderly population, and has symptoms ranging from liver steatosis to advanced fibrosis. Citrus peel extracts (CPEs) contain compounds that potentially improve dyslipidemia; however, the mechanism of action and effects on hepatic steatosis regulation remains unclear. Current study was aimed to investigate the protective effect of CPEs extracted through hot-air drying (CPEW) and freeze-drying (CPEF) and the underlying mechanism in a rat model of high-fat diet-induced NAFLD. The high-fat diet (HFD)-fed rats showed significant increase in total cholesterol, alanine aminotransferase (ALT), triglycerides, aspartate aminotransferase (AST), and lipid peroxidation compared to the normal chow-diet (NCD) group rats; but CPEW and CPEF limited this effect. CPEW and CPEF supplementation reduced both hepatocyte steatosis and fat accumulation involving the regulatory effect of mTORC1. Collectively, CPEW and CPEF protected deterioration of liver steatosis with AMPK activation and regulating ROS accumulation associated with interstitial disorders, which are also associated with endoplasmic reticulum (ER) redox. Thus, the application of CPEW and CPEF may lead to the development of novel therapeutic or preventive agents against NAFLD.


Lifetime Benefits and Harms of Prostate-Specific Antigen-Based Risk-Stratified Screening for Prostate Cancer.
Studies conducted in Swedish populations have shown that men with lowest prostate-specific antigen (PSA) levels at ages 44-50 years and 60 years have very low risk of future distant metastasis or death from prostate cancer. This study investigates benefits and harms of screening strategies stratified by PSA levels. PSA levels and diagnosis patterns from two microsimulation models of prostate cancer progression, detection, and mortality were compared against results of the Malmö Preventive Project, which stored serum and tracked subsequent prostate cancer diagnoses for 25 years. The models predicted the harms (tests and overdiagnoses) and benefits (lives saved and life-years gained) of PSA-stratified screening strategies compared with biennial screening from age 45 years to age 69 years. Compared with biennial screening for ages 45-69 years, lengthening screening intervals for men with PSA less than 1.0 ng/mL at age 45 years led to 46.8-47.0% fewer tests (range between models), 0.9-2.1% fewer overdiagnoses, and 3.1-3.8% fewer lives saved. Stopping screening when PSA was less than 1.0 ng/mL at age 60 years and older led to 12.8-16.0% fewer tests, 5.0-24.0% fewer overdiagnoses, and 5.0-13.1% fewer lives saved. Differences in model results can be partially explained by differences in assumptions about the link between PSA growth and the risk of disease progression. Relative to a biennial screening strategy, PSA-stratified screening strategies investigated in this study substantially reduced the testing burden and modestly reduced overdiagnosis while preserving most lives saved. Further research is needed to clarify the link between PSA growth and disease progression.


Exonic mutations in the SLC12A3 gene cause exon skipping and premature termination in Gitelman syndrome.
A variety of genetic backgrounds cause the loss of function of thiazide-sensitive sodium chloride cotransporter, encoded by SLC12A3, responsible for the phenotypes in Gitelman syndrome. Recently, the phenomenon of exon skipping, in which exonic mutations result in abnormal splicing, has been associated with various diseases. Specifically, mutations in exonic splicing enhancer (ESE) sequences can promote exon skipping. Here, we used a bioinformatics program to analyze 88 missense mutations in the SLC12A3 gene and identify candidate mutations that may induce exon skipping. The three candidate mutations that reduced ESE scores the most were further investigated by minigene assay, and two (p.A356V and p.M672I) caused abnormal splicing in vitro. Furthermore, we identified the p.M672I (c.2016G>A) mutation in a patient with Gitelman syndrome and found that this single nucleotide mutation causes exclusion of exon 16 in the SLC12A3 mRNA transcript. Functional analyses revealed that the protein encoded by the aberrant SLC12A3 transcript does not transport sodium. These results suggest that aberrant exon skipping is one previously unrecognized mechanism by which missense mutations in SLC12A3 can lead to Gitelman syndrome.


Ciprofloxacin in combination with bacteriophage cocktails against multi-drug resistant Pseudomonas aeruginosa in ex vivo simulated endocardial vegetation models.
Pseudomonas aeruginosa-associated infective endocarditis represents difficult-to-treat, deep-seated infections. Phage-antibiotic combinations have shown to eradicate multi-drug resistant (MDR) P. aeruginosa, limit the development of phage resistance, and restore antibiotic sensitivity. The objective of this study was to evaluate the activity of phage-ciprofloxacin (CIP) combinations in 4-day ex vivo simulated endocardial vegetation (SEV) models against drug-resistant P. aeruginosa isolates. Two P. aeruginosa isolates, extensively drug-resistant AR351 and MDR I0003-1, were selected for their drug resistance and sensitivity to phage. Three phages [LL-5504721-AH (LL), E2005-C (EC), and 109] and CIP were evaluated alone and in combination for their activity and influence on drug and phage resistance using 24-h time-kill analysis. The three-phage cocktail (q24h) in combination with CIP (400 mg q12h) was then tested in dynamic 4-day ex vivo SEV models, with reduction of log10 CFU/mL compared using ANOVA with Bonferroni analysis. Compared to other combinations, CIP-LL-EC-109 demonstrated synergistic and bactericidal activity from starting CFU/g against AR351 and I0003-1 (-Δ5.65 and 6.60 log10 CFU/g, respectively; P < 0.001). Additionally, CIP-LL-EC-109 mitigated phage resistance, while all other therapies had a high degree of resistance to >1 phages, and all phage-containing regimens prevented CIP mean inhibitory concentration increases compared to CIP alone for both AR351 and I0003-1 at 96 h.


Developing child mental health services in resource-poor countries.
Despite significant gains in tackling the major causes of child mortality and evidence of an urgent need for child mental health services, resource-poor countries continue to lag behind in child and adolescent mental health service development. This paper analyses possible barriers to the development of child mental health services in resource-poor countries and attempts to proffer solutions. Obstacles identified are the magnitude of child mental health problems that remain invisible to policy makers, an absence of child mental policies to guide the process of service development, and overburdened child mental health professionals. The belief systems about mental illness also prompt help seeking in alternative health systems, thereby reducing the evidence for the burden associated with health seeking. Solutions that may support child mental health service development are the provision of adequate advocacy tools to reveal the burden, poverty alleviation, health awareness programmes, enforcing legislation, training centred within the region, and partnerships with professionals in developed countries. These solutions require simultaneous approaches to encourage service development and utilization. Reductions in child mortality in resource-poor countries will be even more dramatic in the years to come and preparations need to be made to take care of the mental health needs of the children who will survive.


Direct and rapid genotyping of glutathione-S-transferase M1 and T1 from human blood specimens using the SmartAmp2 method.
Clinical studies have suggested that a defect in both glutathione S-transferase (GST) M1 and GSTT1 increases the risk of drug-induced hepatotoxicity. The present study developed the method that enables genotyping of GSTM1 and GSTT1 directly using a small aliquot of blood samples based on an isothermal Smart amplification process version 2 (SmartAmp-2). SmartAmp-2 reaction could complete the genotyping of GSTM1 and GSTT1 within 40 min. The frequency of wild-type, GSTM1 null, GSTT1 null, and double null was 24, 21, 35, and 19%, respectively, consistent with previous reports in the Japanese population. The genotypes of 94 human genomic DNA samples determined by SmartAmp-2 were identical to those determined by the conventional polymerase chain reaction method. SmartAmp-2 was able to determine the genotypes of GSTM1 and GSTT1 even when human blood specimens were used. The SmartAmp-2 method is a rapid and accurate means of identifying the GSTM1 and GSTT1 genotypes, making it less time and more labor efficient in clinical practice than conventional methods requiring preparation of genomic DNA and electrophoresis. This will contribute to evaluate the susceptibility of disease and adverse reactions to drugs caused by deletion of GSTM1 and GSTT1.


Efficient method for optimal placing of water quality monitoring stations for an ungauged basin.
A core problem in monitoring water quality of a river basin is identifying an optimal positioning of a limited number of water-sampling sites. Various optimality criteria have been suggested for this selection process in earlier studies. However, the search for sets of sampling sites that satisfy such criteria poses a challenging optimization problem, especially for a large basin. Here, we show that for particular types of objective functions, the optimization procedure can be dramatically simplified via an analogy with the formulation of Shannon entropy. On this basis, we propose an efficient algorithm that can easily determine the optimal location of water quality sampling sites in a river network. The proposed algorithm can be used standalone or in conjunction with a heuristic optimization algorithm such as a genetic algorithm. For the latter, the proposed algorithm filters only competitive candidates and makes a contribution to reducing the problem size significantly. The superior performance of the proposed method is demonstrated via its application to actual river networks examined in earlier studies, in which the proposed method determines more optimal solutions in a shorter computation time. The idea presented in this study can also be applied to other problems in which the objective function can be formulated in a similar functional form.


Strain variation in phagocytosis of Cryptococcus neoformans: dissociation of susceptibility to phagocytosis from activation and binding of opsonic fragments of C3.
Phagocytosis of Cryptococcus neoformans is markedly influenced by the presence of a polysaccharide capsule. We examined activation and binding of C3 fragments to eight isolates of C. neoformans. All isolates were shown to have capsules by light and electron microscopy. These strains differed in susceptibility to phagocytosis by neutrophils. Yeast cells were opsonized by incubation in normal human serum. Five strains were resistant to ingestion, two strains showed intermediate levels of resistance to ingestion, and one strain was quite sensitive to phagocytosis. Yeast cells opsonized with heat-inactivated serum (56 degrees C for 30 min) neither attached nor were ingested by neutrophils. A quantitative estimate of the amount of C3 bound to the yeast cells was determined by use of normal human serum containing 125I-labeled C3. The results showed approximately 5 X 10(6) to 10 X 10(6) C3 molecules per yeast cell regardless of whether the yeast cells were sensitive or resistant to phagocytosis. Bound C3 was eluted from the yeast cells by treatment with 0.1 M NH2OH (pH 10), and the eluted fragments were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Results of this analysis showed that little of the C3 was in the form of C3b, and there was substantial decay to iC3b, the inactive decay product of C3b. This pattern of decay was similar with all strains. Immunoelectron microscopy was used to assess the ultrastructural location of the C3 fragments bound to the yeast cells. C3 fragments were bound to the perimeter of the capsule regardless of whether the isolate was sensitive or resistant to phagocytosis. Thus, phagocytosis-sensitive and phagocytosis-resistant isolates were similar with regard to the amount, molecular form, and ultrastructural location of C3 fragments bound to the cryptococcal capsule. These results further indicate that activation of the complement cascade is necessary but not sufficient for phagocytosis of the yeast cell.


Identification of novel mutations responsible for resistance to MK-2048, a second-generation HIV-1 integrase inhibitor.
MK-2048 represents a prototype second-generation integrase strand transfer inhibitor (INSTI) developed with the goal of retaining activity against viruses containing mutations associated with resistance to first-generation INSTIs, raltegravir (RAL) and elvitegravir (EVG). Here, we report the identification of mutations (G118R and E138K) which confer resistance to MK-2048 and not to RAL or EVG. These mutations were selected in vitro and confirmed by site-specific mutagenesis. G118R, which appeared first in cell culture, conferred low levels of resistance to MK-2048. G118R also reduced viral replication capacity to approximately 1% that of the isogenic wild-type (wt) virus. The subsequent selection of E138K partially restored replication capacity to approximately 13% of wt levels and increased resistance to MK-2048 to approximately 8-fold. Viruses containing G118R and E138K remained largely susceptible to both RAL and EVG, suggesting a unique interaction between this second-generation INSTI and the enzyme may be defined by these residues as a potential basis for the increased intrinsic affinity and longer "off" rate of MK-2048. In silico structural analysis suggests that the introduction of a positively charged arginine at position 118, near the catalytic amino acid 116, might decrease Mg(2+) binding, compromising enzyme function and thus leading to the significant reduction in both integration and viral replication capacity observed with these mutations.


Evaluating the efficiency of isotope transmission for improved panel design and a comparison of the detection sensitivities of mass cytometer instruments.
The recent introduction of mass cytometry, a technique coupling a cell introduction system generating a stream of single cells with mass spectrometry, has greatly increased the number of parameters that can be measured per single cell. As with all new technology there is a need for dissemination of standardization and quality control procedures. Here, we characterize variations in sensitivity observed across the mass range of a mass cytometer, using different lanthanide tags. We observed a five-fold difference in lanthanide detection over the mass range and demonstrated that each instrument has its own sensitivity pattern. Therefore, the selection of lanthanide combinations is a key step in the establishment of a staining panel for mass cytometry-based experiments, particularly for multicenter studies. We propose the sensitivity pattern as the basis for panel design, instrument standardization and future implementation of normalization algorithms.


Parental education level and adolescent depression: A multi-country meta-analysis.
Adolescent depression, as a common problem, has always been the focus of attention and research. However, no consistent conclusion has been drawn on its relationship with parental education level. Therefore, this study used meta-analysis and dose-response analysis techniques to explore the overall relationship between the two, and further explore the moderating factors affecting the relationship between the two through subgroup analysis and meta-regression analysis. After a literature search and screening, a total of 22 literatures were included, including 634,821 adolescents aged 9-25, distributed in 10 countries. The results showed that: (1) Overall parental education level was negatively correlated with adolescent depressive symptoms (RR = 0.88.p = 0.000), and there was a dose-response relationship between the two. (2) Geographical location (p = 0.002 < 0.05) and national economic development (p = 0.03 < 0.05) significantly moderated and affected the association between parental education level and adolescent depression, thus presenting a global inconsistency. The results of this study provide a more accurate conclusion on the relationship between parental education and adolescent depression to some extent, highlight the importance and necessity of considering family, economic and cultural factors when studying and solving adolescent depression problems, and guide us to pay more attention to the cross-regional and cross-cultural differences in adolescent depression problems.