Distinct alkaline phosphatase in serum of patients with lymphatic leukemia and infectious mononucleosis.
A distinct alkaline phosphatase (phosphatase N) was demonstrated in the serum of patients with acute lymphatic leukemia, chronic lymphatic leukemia, and infectious mononucleosis. This enzyme closely resembles that extracted from the thymus of mice with lymphoma or lymphatic leukemia, both in its electro-phoretic mobility and its substrate specificity. The phosphatase N activity was related to the clinical state of patients with lymphatic leukemia and disappeared with recovery from infectious mnononucleosis.


Structural Basis of Catalysis in the Bacterial Monoterpene Synthases Linalool Synthase and 1,8-Cineole Synthase.
Terpenoids form the largest and stereochemically most diverse class of natural products, and there is considerable interest in producing these by biocatalysis with whole cells or purified enzymes, and by metabolic engineering. The monoterpenes are an important class of terpenes and are industrially important as flavors and fragrances. We report here structures for the recently discovered Streptomyces clavuligerus monoterpene synthases linalool synthase (bLinS) and 1,8-cineole synthase (bCinS), and we show that these are active biocatalysts for monoterpene production using biocatalysis and metabolic engineering platforms. In metabolically engineered monoterpene-producing E. coli strains, use of bLinS leads to 300-fold higher linalool production compared with the corresponding plant monoterpene synthase. With bCinS, 1,8-cineole is produced with 96% purity compared to 67% from plant species. Structures of bLinS and bCinS, and their complexes with fluorinated substrate analogues, show that these bacterial monoterpene synthases are similar to previously characterized sesquiterpene synthases. Molecular dynamics simulations suggest that these monoterpene synthases do not undergo large-scale conformational changes during the reaction cycle, making them attractive targets for structured-based protein engineering to expand the catalytic scope of these enzymes toward alternative monoterpene scaffolds. Comparison of the bLinS and bCinS structures indicates how their active sites steer reactive carbocation intermediates to the desired acyclic linalool (bLinS) or bicyclic 1,8-cineole (bCinS) products. The work reported here provides the analysis of structures for this important class of monoterpene synthase. This should now guide exploitation of the bacterial enzymes as gateway biocatalysts for the production of other monoterpenes and monoterpenoids.


[Autopsies and surgical service].
The aim of autopsy is to define the causes of a patient's death by studying the gross and microscopic visceral lesions. It is in actual decline but nevertheless the post-mortem examination remains one of the basic tools for the assessment of medical care in hospital. The aim of this paper is to finalize the present French legislation of autopsy and to show its importance in surgical practice.


[Werdnig-Hoffmann's congenital spinal atrophy in nursing infants].
The paper is concerned with an analysis of the anamnestic clinical and histological data on 8 children (6 girls and 2 boys) who died at an age of 3 to 7 months. They had been treated at the Child's Clinical Hospital of Moscow for Hoffmann-Werdnig syndrome, bilateral polysegmental pneumonia, respiratory insufficiency, stage III, hypotrophy, stages II and III, anemia of infectious alimentary etiology. Ground for hospitalization was superaddition of intercurrent infection, the growth of respiratory insufficiency. In spite of intensive care, the children died on days I-III of the stay at the hospital. The clinical analysis of the disease and the autopsy findings allowed a conclusion about the predominance of the early form of Hoffmann-Werdnig syndrome running a rapid progressive course. The immediate cause of the children's death was acute respiratory insufficiency associated with polysegmental pneumonia, spinal amyotrophy.


Effect of Proinflammatory S100A9 Protein on Migration and Proliferation of Microglial Cells.
Alzheimer's disease (AD) is a multifactorial disease affecting aging population worldwide. Neuroinflammation became a focus of research as one of the major pathologic processes relating to the disease onset and progression. Proinflammatory S100A9 is the central culprit in the amyloid-neuroinflammatory cascade implicated in AD and other neurodegenerative diseases. We studied the effect of S100A9 on microglial BV-2 cell proliferation and migration. The responses of BV-2 cells to S100A9 stimulation were monitored in real-time using live cell microscopy, transcriptome sequencing, immunofluorescence staining, western blot analysis, and ELISA. We observed that a low dose of S100A9 promotes migration and proliferation of BV-2 cells. However, acute inflammatory condition (i.e., high S100A9 doses) causes diminished cell viability; it is uncovered that S100A9 activates TLR-4 and TLR-7 signaling pathways, leading to TNF-α and IL-6 expression, which affect BV-2 cell migration and proliferation in a concentration-dependent manner. Interestingly, the effects of S100A9 are not only inhibited by TNF-α and IL-6 antibodies. The addition of amyloid-β (Aβ) 1-40 peptide resumes the capacities of BV-2 cells to the level of low S100A9 concentrations. Based on these results, we conclude that in contrast to the beneficial effects of low S100A9 dose, high S100A9 concentration leads to impaired mobility and proliferation of immune cells, reflecting neurotoxicity at acute inflammatory conditions. However, the formation of Aβ plaques may be a natural mechanism that rescues cells from the proinflammatory and cytotoxic effects of S100A9, especially considering that inflammation is one of the primary causes of AD.


Spectroscopy of single- and double-wall carbon nanotubes in different environments.
Individual single-wall carbon nanotubes (SWNTs) and double-wall carbon nanotubes (DWNTs) were suspended in water for optical studies using sodium-cholate and other surfactants. We used time-resolved photoluminescence (PL) spectroscopy to study the influence of tube chirality and diameter as well as of the environment on nonradiative decay in small diameter tubes. The studies provide evidence for PL from small diameter core tubes in DWNTs and for a correlation of nonradiative decay with tube diameter and exciton red shift as induced by interaction with the environment.


"Visceral Consciousness": The Gut-Brain Axis in Sleep and Sleeplessness in Britain and America, 1850-1914.
Sleeplessness was a quotidian yet challenging problem for medical practitioners in Britain and America in the nineteenth and early twentieth centuries. While physiologists endeavored to unravel the secrets of sleep by examining the brain, in the clinic doctors looked to the gut as a site through which sleeplessness was both caused and cured. This article explores the gut-brain axis in medical literature on sleep and sleep loss in this period. It argues that despite the lack of a coherent understanding of the gut-brain connection, the digestive system was central to how physiologists and clinicians approached sleeplessness. It employs Victorian physician Joseph Mortimer Granville's (1833-1900) concept of "visceral consciousness" to better understand the varied and often contradictory explanatory constellations that emerged to elucidate the role of digestion in sleeplessness.


Effects of PEG antibodies on in vivo performance of LNP-mRNA vaccines.
Polyethylene glycol (PEG) plays important roles in stabilizing and lengthening circulation time of lipid nanoparticle (LNP) vaccines. Nowadays various levels of PEG antibodies have been detected in human blood, but the impact and mechanism of PEG antibodies on the in vivo performance of LNP vaccines has not been clarified thoroughly. By illustrating the distribution characteristics of PEG antibodies in human, the present study focused on the influence of PEG antibodies on the safety and efficacy of LNP-mRNA vaccine against COVID-19 in animal models. It was found that PEG antibodies led to shortened blood circulation duration, elevated accumulation and mRNA expression in liver and spleen, enhanced expression in macrophage and dendritic cells, while without affecting the production of anti-Spike protein antibodies of COVID-19 LNP vaccine. Noteworthily, PEG antibodies binding on the LNP vaccine increased probability of complement activation in animal as well as in human serum and led to lethal side effect in large dosage via intravenous injection of mice. Our data suggested that PEG antibodies in human was a risky factor of LNP-based vaccines for biosafety concerns but not efficacy.


The SNAP-25 linker as an adaptation toward fast exocytosis.
The assembly of four soluble N-ethylmaleimide-sensitive factor attachment protein receptor domains into a complex is essential for membrane fusion. In most cases, the four SNARE-domains are encoded by separate membrane-targeted proteins. However, in the exocytotic pathway, two SNARE-domains are present in one protein, connected by a flexible linker. The significance of this arrangement is unknown. We characterized the role of the linker in SNAP-25, a neuronal SNARE, by using overexpression techniques in synaptosomal-associated protein of 25 kDa (SNAP-25) null mouse chromaffin cells and fast electrophysiological techniques. We confirm that the palmitoylated linker-cysteines are important for membrane association. A SNAP-25 mutant without cysteines supported exocytosis, but the fusion rate was slowed down and the fusion pore duration prolonged. Using chimeric proteins between SNAP-25 and its ubiquitous homologue SNAP-23, we show that the cysteine-containing part of the linkers is interchangeable. However, a stretch of 10 hydrophobic and charged amino acids in the C-terminal half of the SNAP-25 linker is required for fast exocytosis and in its absence the calcium dependence of exocytosis is shifted toward higher concentrations. The SNAP-25 linker therefore might have evolved as an adaptation toward calcium triggering and a high rate of execution of the fusion process, those features that distinguish exocytosis from other membrane fusion pathways.


The efficiency analysis of thrombolytic rt-PA combined with intravascular interventional therapy in patients with acute basilar artery occlusion.
In order to further optimize the treatment strategy for the patients with acute basilar artery occlusion, we were dedicated to study the therapeutic effects and influential factors in the process of treated basilar artery occlusion with thrombolytic combined vascular interventional therapy. 75 patients with acute basilar artery occlusion treated with arterial thrombolytic therapy were analyzed retrospectively. In accordance with the discharge records of patients, their short-term curative effect with 24-hour treatment and 14-days treatment were evaluated. Our data showed that the survival condition of the patients with acute acute basilar artery occlusion were visibly improved by combination thrombolytic and interventional therapy. Moreover, their BI scores were remarkably improved, while NIHSS and mRS scores were evidently reduced. These data proved that our treatment strategy was able to improve the survival condition of patients with acute basilar artery occlusion. Furthermore, our data showed that coagulation related factors remarkably improved in the patients, when they treated by combination thrombolytic therapy with interventional therapy. In addition, our results suggested that the patients' bilateral Babinski(+), revascularization and coma symptom were closely related to their prognosis after treated the patients with combination thrombolytic and vascular interventional therapy, and the difference was statistically significant (p<0.05, p<0.05, p<0.05). Besides, our data also displayed that the with stent assisted angioplasty was significantly superior to the patients with balloon angioplasty, and the difference was statistically significant (p<0.05). Anyhow, combination thrombolytic with interventional therapy can effectively promote the prognosis of the patients with acute basilar artery occlusion. The coma symptom, bilateral Babinski(+), and revascularization in the patients with acute basilar artery occlusion have an appreciable impact on the patients' prognosis.


Risk factors for psychosocial dysfunction among enrollees in the State Children's Health Insurance Program.
The authors studied enrollees in the State Children's Health Insurance Program (SCHIP) (Title XXI) to characterize risk factors for psychosocial dysfunction among children of the working poor. Medical and psychosocial variables were included in a survey completed by 393 parents of children enrolled in SCHIP. Regression analysis was used to examine the relationship between these variables and scores on the Pediatric Symptom Checklist, a measure of psychosocial dysfunction among children. Stepwise multiple regression showed that parental dysphoria, parental history of psychiatric or substance use problems, childhood chronic medical illness, and exposure to traumatic events each contributed independently to variance in psychosocial dysfunction in this population, explaining 34 percent of total variance. Despite strong progress in implementing SCHIP at the state level, the behavioral health care needs of children of the working poor have not been well defined. This study identified risk factors that can be easily found in the patient's medical record or detected during an interview by the primary care physician. Thus screening to identify children at risk of psychosocial dysfunction is warranted among SCHIP enrollees.


Role of sex in the neurochemical and neuroendocrine correlates of paw preference in the rat.
Handedness in humans and paw preference (PP) in rodents have been associated with neural and physiological correlates, which frequently appear to be sex-specific. The present study examines sex differences in the effects of differential PP on post mortem measures of regional monoamine activity in adult Long-Evans rats. The effects of PP on neuroendocrine function were also assessed by measuring plasma adrenocorticotropic hormone (ACTH) in response to a 30-min restraint stress. Most rats showed strong individual PP. Males (n=27) and females (n=26) did not differ in the direction or strength of their PP with nearly equal numbers of left and right-pawed rats. However, many Sex×PP interactions were noted in regional neurochemical measures, and most effects of PP were bilateral in nature and sex-specific. In males, Left PP (relative to Right PP) was associated with reduced striatal dopamine (DA) levels and amygdala DA metabolism bilaterally. In females, Left PP was associated with a bilateral upregulation of DAergic metabolism in both prefrontal cortex and nucleus accumbens, increased amygdala serotonin metabolism, and a (right) unilateral increase in amygdala DA. As in previous studies, some correlations were also noted between PP and asymmetrical or unilateral monoamine measures. As well, rats with strong PP, independent of direction or sex, had lower basal ACTH and more robust stress responses than rats with weak PP, suggesting a possible adaptive advantage to strong lateralization. Overall, sex differences were found only regarding effects of the direction of PP, not its magnitude. The findings suggest that the direction of PP affects distinct bilateral networks of structures sex-dependently. Such fundamental influences of PP on functional brain organization have implications for a wide array of processes under monoaminergic modulation in these brain regions, and may further our understanding of the numerous human examples of gender and handedness interactions across several modalities.


An autopsy-proven case of myeloperoxidase-antineutrophil cytoplasmic antibody-positive polyarteritis nodosa with acute renal failure and alveolar hemorrhage.
An 80-year-old woman positive for myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA) was admitted with a 3-month history of fever, general malaise, and weight loss, after unsuccessful treatment with antibiotics. Upon admission, her fever persisted, and there was concomitant deterioration of renal function without active urine sediments. Furthermore, she developed hemoptysis, and chest computed tomography (CT) scan revealed bilateral diffuse alveolar hemorrhage. Although a renal biopsy was not performed because of her dementia, we initially suspected microscopic polyangiitis (MPA) on the basis of her clinical course. Because of her poor general condition, she was administered a low dose of prednisolone. Although her fever subsided, she suffered from intractable alveolar hemorrhage and eventually died from respiratory failure. During the autopsy, fibrinoid necrosis was restricted to medium-sized arteries, including the arcuate arteries of the kidneys and the bronchial arteries, without necrotizing crescentic glomerulonephritis and alveolar capillaritis. Therefore, polyarteritis nodosa (PAN) was diagnosed. It is important to distinguish between MPA and PAN because they can lead to life-threatening complications, and their treatment strategies and prognosis are different. When a patient presents with MPO-ANCA, alveolar hemorrhage, and acute renal failure with little evidence of glomerulonephritis, a differential diagnosis of PAN should be made; however, it is difficult to do so without pathological findings. Therefore, pathological examination should be carried out whenever possible.


Expression, Purification, and Characterisation of Dehydroquinate Synthase from Pyrococcus furiosus.
Dehydroquinate synthase (DHQS) catalyses the second step of the shikimate pathway to aromatic compounds. DHQS from the archaeal hyperthermophile Pyrococcus furiosus was insoluble when expressed in Escherichia coli but was partially solubilised when KCl was included in the cell lysis buffer. A purification procedure was developed, involving lysis by sonication at 30°C followed by a heat treatment at 70°C and anion exchange chromatography. Purified recombinant P. furiosus DHQS is a dimer with a subunit Mr of 37,397 (determined by electrospray ionisation mass spectrometry) and is active over broad pH and temperature ranges. The kinetic parameters are K(M) (3-deoxy-D-arabino-heptulosonate 7-phosphate) 3.7 μM and k(cat) 3.0 sec(-1) at 60°C and pH 6.8. EDTA inactivates the enzyme, and enzyme activity is restored by several divalent metal ions including (in order of decreasing effectiveness) Cd(2+), Co(2+), Zn(2+), and Mn(2+). High activity of a DHQS in the presence of Cd(2+) has not been reported for enzymes from other sources, and may be related to the bioavailability of Cd(2+) for P. furiosus. This study is the first biochemical characterisation of a DHQS from a thermophilic source. Furthermore, the characterisation of this hyperthermophilic enzyme was carried out at elevated temperatures using an enzyme-coupled assay.


Prenatal methylazoxymethanol treatment in rats produces brain abnormalities with morphological similarities to human developmental brain dysgeneses.
A double methylazoxymethanol (MAM) intraperitoneal injection was prenatally administered to pregnant rats at gestational day 15 to induce developmental brain dysgeneses. Thirty adult rats from 8 different progenies were investigated with a combined electrophysiological and neuroanatomical analysis. The offspring of treated dams was characterized by extensive cortical layering abnormalities, subpial bands of heterotopic neurons in layer I, and subcortical nodules of heterotopic neurons extending from the periventricular region to the hippocampus and neocortex. The phenotype of cell subpopulations within the heterotopic structures was analyzed by means of antibodies raised against glial and neuronal markers, calcium binding proteins, GABA, and AMPA glutamate receptors. Neurons within the subcortical heterotopic nodules were characterized by abnormal firing properties, with sustained repetitive bursts of action potentials. The subcortical nodules were surrounded by cell clusters with ultrastructural features of young migrating neurons. The immunocytochemical data suggested, moreover, that the subcortical heterotopia were formed by neurons originally committed to the neocortex and characterized by morphological features similar to those found in human periventricular nodular heterotopia. The present study demonstrates that double MAM treatment at gestational day 15 induces in rats developmental brain abnormalities whose anatomical and physiological features bear resemblance to those observed in human brain dysgeneses associated with intractable epilepsy. Therefore, MAM treated rats could be considered as useful tools in investigating the pathogenic mechanisms involved in human developmental brain dysgeneses.


School dental screening programmes for oral health.
School dental screening refers to visual inspection of children's oral cavity in a school setting followed by making parents aware of their child's current oral health status and treatment needs. Screening at school intends to identify children at an earlier stage than symptomatic disease presentation, hence prompting preventive and therapeutic oral health care for the children. This review evaluates the effectiveness of school dental screening in improving oral health status. It is an update of the original review, which was first published in December 2017. To assess the effectiveness of school dental screening programmes on overall oral health status and use of dental services. Cochrane Oral Health's Information Specialist searched the following databases: Cochrane Oral Health's Trials Register (to 4 March 2019), the Cochrane Central Register of Controlled Trials (CENTRAL, the Cochrane Register of Studies, to 4 March 2019), MEDLINE Ovid (1946 to 4 March 2019), and Embase Ovid (15 September 2016 to 4 March 2019). The US National Institutes of Health Trials Registry (ClinicalTrials.gov) and the World Health Organization International Clinical Trials Registry Platform were searched for ongoing trials. No restrictions were placed on language or publication status when searching the electronic databases; however, the search of Embase was restricted to the last six months due to the Cochrane Centralised Search Project to identify all clinical trials and add them to CENTRAL. We included randomised controlled trials (RCTs) (cluster or parallel) that evaluated school dental screening compared with no intervention or with one type of screening compared with another. We used standard methodological procedures expected by Cochrane. We included seven trials (five were cluster-RCTs) with 20,192 children who were 4 to 15 years of age. Trials assessed follow-up periods of three to eight months. Four trials were conducted in the UK, two were based in India and one in the USA. We assessed two trials to be at low risk of bias, two trials to be at high risk of bias and three trials to be at unclear risk of bias.None of the trials had long-term follow-up to ascertain the lasting effects of school dental screening.None of the trials reported the proportion of children with untreated caries or other oral diseases, cost effectiveness or adverse events.Four trials evaluated traditional screening versus no screening. We performed a meta-analysis for the outcome 'dental attendance' and found an inconclusive result with high heterogeneity. The heterogeneity was found to be, in part, due to study design (three cluster-RCTs and one individual-level RCT). Due to the inconsistency, we downgraded the evidence to 'very low certainty' and are unable to draw conclusions about this comparison.Two cluster-RCTs (both four-arm trials) evaluated criteria-based screening versus no screening and showed a pooled effect estimate of RR 1.07 (95% CI 0.99 to 1.16), suggesting a possible benefit for screening (low-certainty evidence). There was no evidence of a difference when criteria-based screening was compared to traditional screening (RR 1.01, 95% CI 0.94 to 1.08) (very low-certainty evidence).In one trial, a specific (personalised) referral letter was compared to a non-specific one. Results favoured the specific referral letter with an effect estimate of RR 1.39 (95% CI 1.09 to 1.77) for attendance at general dentist services and effect estimate of RR 1.90 (95% CI 1.18 to 3.06) for attendance at specialist orthodontist services (low-certainty evidence).One trial compared screening supplemented with motivation to screening alone. Dental attendance was more likely after screening supplemented with motivation, with an effect estimate of RR 3.08 (95% CI 2.57 to 3.71) (low-certainty evidence).Only one trial reported the proportion of children with treated dental caries. This trial evaluated a post screening referral letter based on the common-sense model of self-regulation (a theoretical framework that explains how people understand and respond to threats to their health), with or without a dental information guide, compared to a standard referral letter. The findings were inconclusive. Due to high risk of bias, indirectness and imprecision, we assessed the evidence as very low certainty. The trials included in this review evaluated short-term effects of screening. We found very low-certainty evidence that is insufficient to allow us to draw conclusions about whether there is a role for traditional school dental screening in improving dental attendance. For criteria-based screening, we found low-certainty evidence that it may improve dental attendance when compared to no screening. However, when compared to traditional screening, there is no evidence of a difference in dental attendance (very low-certainty evidence).We found low-certainty evidence to conclude that personalised or specific referral letters may improve dental attendance when compared to non-specific counterparts. We also found low-certainty evidence that screening supplemented with motivation (oral health education and offer of free treatment) may improve dental attendance in comparison to screening alone. For children requiring treatment, we found very-low certainty evidence that was inconclusive regarding whether or not a referral letter based on the 'common-sense model of self-regulation' was better than a standard referral letter.We did not find any trials addressing possible adverse effects of school dental screening or evaluating its effectiveness for improving oral health.


Neurologists--1990.
The American Medical Association Physician Masterfile for 1990 contains 13,705 physicians who consider neurology their primary, secondary, or tertiary specialty. Of these, 7,036 are board certified and 10,065 designated neurology as their primary specialty. Although there has been a steady increase in the number of women, the vast majority of neurologists are men and less than 50 years old. Two-thirds provide direct patient care as their major activity and 15% are in training. There are almost as many neurologists employed by hospitals, medical schools, and government entities (40%) as there are working alone or in group practice (48%). The actual number of neurologists is compared with earlier projections of neurology needs for 1990. Since forecasts, in the absence of more detailed information, can become the reality of public policy decisions, neurologists will need to become increasingly informed about the many unique benefits they provide to their patients.


[Perniosis and lupus anticoagulant].
Five patients with acral chilblains and lupus anticoagulant activity revealed by coagulation analysis are reported. Three patients suffered a systemic lupus erythematosus, and two of them a completely developed antiphospholipid syndrome as well. Another case had chronic cutaneous lupus with only facial discoid lesions, and in the last one the lupus anticoagulant was likely related to a chronic liver infection of hepatitis B virus because she had not erythematous lupus. There are few dermatologic references about chilblain associated to lupus anticoagulant. Certain evidences suggest a pathogenic relation of this findings.


Comparison of patella cartilage volume and radiography in the assessment of longitudinal joint change at the patellofemoral joint.
To compare radiological assessment of the patellofemoral (PF) joint and cartilage volume as measured by magnetic resonance imaging (MRI) in a longitudinal study. One hundred and two subjects with osteoarthritis (OA) had baseline and followup skyline and lateral radiographs and MRI of the same knee. Duration of followup was 1.94 +/- 0.23 years. Mid-patella joint space and narrowest joint space were measured on lateral PF radiographs. Minimum joint space in the lateral and medial facets was measured on skyline radiographs. Rate of progression of PF joint OA was measured quantitatively at each site. Patella cartilage volume was measured from the MRI. The rate of change in each radiographic measure was compared to the rate of change in patella cartilage volume. The average loss of patella cartilage was 133 +/- 143 microm(3)/yr (4.4%). The loss of joint space over the same period on lateral radiographs was 0.7 +/- 2.6 mm at the narrowest joint space and 0.2 +/- 3.8 mm at the mid-patella joint space. On the skyline radiographs, the loss of joint space at the medial facet was 0.4 +/- 3.9 mm and 0.2 +/- 3.4 mm at the lateral facet. Only change in narrowest joint space on the lateral PF radiographs correlated with change in patella cartilage volume (R = 0.22, p = 0.03). Our data suggest that narrowest joint space measured on lateral PF radiographs correlates best with loss of patella cartilage. Further work will be needed to determine how different radiological measures of the PF joint relate to the disease process in OA.


Adaptive spatial control of fiber modes and their excitation for high-harmonic generation.
We present the control of high-harmonic generation (HHG) in hollow fibers using adaptive pulse shaping techniques. The shaping capabilities of our spatial light modulator (SLM) are demonstrated by the excitation of specific fiber modes inside a hollow fiber with a helium-neon laser. Afterwards spatially shaped ultrashort pulses are used to generate phase-matched high-harmonic radiation in a fiber. We show that by controlling the mode structure, we can manipulate the spatial and spectral properties of the generated harmonics.


[Clinical-epidemiologic indications for echocardiographic assessment in the neonatal period. Value of risk groups].
An echocardiographic study was performed in newborns from risk groups, with the aim of to determining prevalence and to evaluate the indications for this test in the neonatal period. One hundred fifty six newborns were studied. They were admitted to the Newborn ward of the Department of Obstetric of the HC-FMUSP, in the period of November 91 to April 93, from mothers with congenital heart disease or diabetes, low birth weight newborns, with extracardiac malformations, cardiac signs and/or congenital infections. The observed prevalence was 21.8%, greater than that of the general population (0.8-1.2%). The group composed by 27 children with extracardiac abnormalities presented the largest prevalence when compared with the other groups (40.7%). These data justify the use of echocardiography in high risk newborns for the detection of congenital heart disease.


Characteristics and Survival Outcomes of Children With Hodgkin Lymphoma Treated Primarily With Chemotherapy.
Hodgkin disease is a malignant tumor of the lymphatic system that comprises ∼6% of childhood cancers. In developing countries, efforts are made to ensure adherence to standard protocol/regimens, study patients' outcomes, and compare with that in developed world. We conducted a retrospective medical records' review of 212 children younger than 20 years presenting to The Indus Hospital in Pakistan with previously untreated Hodgkin lymphoma between August 2000 and December 2012. We collected demographic and other epidemiologic variables such as age, sex, stage, subtype of disease, and survival outcomes. The mean±SD age of patients at time of diagnosis was 9.0±3.8 years with a male to female ratio of ∼4.7:1. In total, 44 (20.8%) patients were 5 years of age or above at presentation. Overall, 131 (61.8%) patients presented with B-symptoms and mixed cellularity was the most frequently diagnosed subtype in 65.1% of cases. In total, 170 (80.2%) achieved full remission after completion of chemotherapy. Patients were treated with alternating cycles of ABVD (Adriamycin, Bleomycin, Vincristine, and Dacarbazine) and COPDAC (Cyclophosphamide, Vincristine, Prednisolone, and Dacarbazine). The majority (n=114, 59.1%) received 6 cycles of chemotherapy, 44 (22.8%) received ≤4 cycles followed by 24 (12.4%) receiving 8 cycles. Radiotherapy was administered only to those patients with significant residual disease at the end of chemotherapy (n=20, 10%). The 5-year overall survival and event-free survival in our cohort was 89.6% and 82.1%, respectively. Our findings suggest that treatment with 4 to 8 alternating cycles of ABVD/COPDAC has an excellent outcome in childhood Hodgkin disease.


Genetically engineered crops and pesticide use in U.S. maize and soybeans.
The widespread adoption of genetically engineered (GE) crops has clearly led to changes in pesticide use, but the nature and extent of these impacts remain open questions. We study this issue with a unique, large, and representative sample of plot-level choices made by U.S. maize and soybean farmers from 1998 to 2011. On average, adopters of GE glyphosate-tolerant (GT) soybeans used 28% (0.30 kg/ha) more herbicide than nonadopters, adopters of GT maize used 1.2% (0.03 kg/ha) less herbicide than nonadopters, and adopters of GE insect-resistant (IR) maize used 11.2% (0.013 kg/ha) less insecticide than nonadopters. When pesticides are weighted by the environmental impact quotient, however, we find that (relative to nonadopters) GE adopters used about the same amount of soybean herbicides, 9.8% less of maize herbicides, and 10.4% less of maize insecticides. In addition, the results indicate that the difference in pesticide use between GE and non-GE adopters has changed significantly over time. For both soybean and maize, GT adopters used increasingly more herbicides relative to nonadopters, whereas adopters of IR maize used increasingly less insecticides. The estimated pattern of change in herbicide use over time is consistent with the emergence of glyphosate weed resistance.


Redefining the Subsurface Biosphere: Characterization of Fungi Isolated From Energy-Limited Marine Deep Subsurface Sediment.
The characterization of metabolically active fungal isolates within the deep marine subsurface will alter current ecosystem models and living biomass estimates that are limited to bacterial and archaeal populations. Although marine fungi have been studied for over fifty years, a detailed description of fungal populations within the deep subsurface is lacking. Fungi possess metabolic pathways capable of utilizing previously considered non-bioavailable energy reserves. Therefore, metabolically active fungi would occupy a unique niche within subsurface ecosystems, with the potential to provide an organic carbon source for heterotrophic prokaryotic populations from the transformation of non-bioavailable energy into substrates, as well as from the fungal necromass itself. These organic carbon sources are not currently being considered in subsurface energy budgets. Sediments from South Pacific Gyre subsurface, one of the most energy-limited environments on Earth, were collected during the Integrated Ocean Drilling Program Expedition 329. Anoxic and oxic sediment slurry enrichments using fresh sediment were used to isolate multiple fungal strains in media types that varied in organic carbon substrates and concentration. Metabolically active and dormant fungal populations were also determined from nucleic acids extracted from in situ cryopreserved South Pacific Gyre sediments. For further characterization of physical growth parameters, two isolates were chosen based on their representation of the whole South Pacific Gyre fungal community. Results from this study show that fungi have adapted to be metabolically active and key community members in South Pacific Gyre sediments and potentially within global biogeochemical cycles.


Susceptibility characterisation of Candida spp. to four essential oils.
In the present investigation, anti-Candida activity of four essential oils i.e. Black cumin (Nigella sativa), Curry leaf (Murraya koienigii), Ajwain (Trachiyspirum ammi), and Betel leaf (Piper betel) were screened against four human pathogenic species of Candida viz. Candida albicans, Candida tropicalis, Candida glabrata, and Candida parapsilosis. The minimum inhibitory concentration (MIC) values of the oils ranged between 15.62 and 250 μl/ml while studied through tube dilution method. The oils retained their anti-Candida activities even after heat treatment (at 45ΊC, 60ΊC, 100ΊC for 1 hour) and also on autoclaving. Both Ajwain and Black Cumin leaf oils showed better anti-Candida activity against Candida albicans, resulting in an irreversible damage to the cells. The anti-Candida activity of these essential oils could be attributable to the membrane inhibition mechanism. The activity of the oils is reported to be microbicidal (Candida-cidal).


In situ STM imaging of the structures of pentacene molecules adsorbed on Au(111).
In situ scanning tunneling microscope (STM) was used to examine the spatial structures of pentacene molecules adsorbed onto a Au(111) single-crystal electrode from a benzene dosing solution containing 16-400 microM pentacene. Molecular-resolution STM imaging conducted in 0.1 M HClO(4) revealed highly ordered pentacene structures of ( radical31 x radical31)R8.9 degrees , (3 x 10), ( radical31 x 10), and ( radical7 x 2 radical7)R19.1 degrees adsorbed on the reconstructed Au(111) electrode dosed with different pentacene solutions. These pentacene structures and the reconstructed Au(111) substrate were stable between 0.2 and 0.8 V [vs reversible hydrogen electrode, RHE]. Increasing the potential to E > 0.8 V lifted the reconstructed Au(111) surface and disrupted the ordered pentacene adlattices simultaneously. Ordered pentacene structures could be restored by applying potentials negative enough to reinforce the reconstructed Au(111). At potentials negative of 0.2 V, the adsorption of protons became increasingly important to displace adsorbed pentacene admolecules. Although the reconstructed Au(111) structure was not essential to produce ordered pentacene adlayers, it seemed to help the adsorption of pentacene molecules in a long-range ordered pattern. At room temperature (25 degrees C), approximately 100 pentacene molecules seen in STM images could rotate and align themselves to a neighboring domain in 10 s, suggesting that pentacene admolecules could be mobile on Au(111) under the STM imaging conditions of -150 mV in bias voltage and 1 nA in feedback current.


Improvement of growth of Chlamydia pneumoniae on HEp-2 cells by pretreatment with polyethylene glycol in combination with additional centrifugation and extension of culture time.
The following adaptations led to improved growth of Chlamydia pneumoniae on HEp-2 cells compared to that by the standard method: monolayer preincubation with 7% polyethylene glycol (PEG), extension of incubation time to 7 days, and extension of incubation to 7 days in combination with centrifugation on days 3, 4, and 5. These adaptations resulted in approximate increases in numbers of inclusion-forming units (IFU) of 2-, 5-, and 69-fold, respectively. A combination of preincubation with PEG, prolonged incubation, and centrifugation on days 3, 4, and 5 increased the numbers of IFU >300-fold. This is therefore recommended as the optimal method for culturing C. pneumoniae.


Use of the kurtosis statistic in an evaluation of the effects of noise and solvent exposures on the hearing thresholds of workers: An exploratory study.
The aim of this exploratory study was to examine whether the kurtosis metric can contribute to investigations of the effects of combined exposure to noise and solvents on human hearing thresholds. Twenty factory workers exposed to noise and solvents along with 20 workers of similar age exposed only to noise in eastern China were investigated using pure-tone audiometry (1000-8000 Hz). Exposure histories and shift-long noise recording files were obtained for each participant. The data were used in the calculation of the cumulative noise exposure (CNE) and CNE adjusted by the kurtosis metric for each participant. Passive samplers were used to measure solvent concentrations for each worker exposed to solvents over the full work shift. Results showed an interaction between noise exposure and solvents for the hearing threshold at 6000 Hz. This effect was observed only when the CNE level was adjusted by the kurtosis metric.


Impairment of eyeblink classical conditioning in progressive supranuclear palsy.
In a previous study we showed that learning in eyeblink classical conditioning (EBCC) is normal in Parkinson's disease (PD) and that the serial reaction time task (SRTT) is only marginally impaired. Since pathological lesions are more widespread in the atypical parkinsonian disorder of progressive supranuclear palsy (PSP) than in PD, we hypothesized that PSP patients may show more profound deficits in the EBCC and SRTT learning tasks. We therefore investigated EBCC with a delay and two trace paradigms, an SRTT and the California Verbal Learning Test (CVLT) in eight patients with PSP and an age-matched control group. In all EBCC paradigms, we found a significant difference between groups with no significant learning in PSP patients. In the SRTT, implicit learning may have been impaired, but verbal and manual sequence recall were only marginally impaired. Verbal memory was significantly worse in PSP patients than in the control group. Our study shows a dissociated pattern of learning abilities in PSP, where the EBCC as a measure of implicit learning is impaired, the explicit sequence detection in the SRTT is relatively preserved, and the verbal memory impaired. We hypothesize that the PSP patients' deficits in EBCC learning may be due to lesions of deep cerebellar nuclei. There may be a clinical role for EBCC in distinguishing PD and PSP patients.


How the Aridification of Australia Structured the Biogeography and Influenced the Diversification of a Large Lineage of Australian Cicadas.
Over the last 30 million years, Australia's landscape has undergone dramatic cooling and drying due to the establishment of the Antarctic Circumpolar Current and change in global CO$_{2}$ levels. Studies have shown that many Australian organisms went extinct during these major cooling events, while others experienced adaptive radiations and increases in diversification rates as a result of exploiting new niches in the arid zone. Despite the many studies on diversification and biogeography in Australia, few have been continent-wide and none have focused on a group of organisms adapted to feeding on plants. We studied 162 species of cicadas in the Australian Pauropsalta complex, a large generic lineage within the tribe Cicadettini. We asked whether there were changes in the diversification rate of Pauropsalta over time and if so: 1) which clades were associated with the rate change? 2) did timing of rate shifts correspond to known periods of dramatic historical climate change, 3) did increases in diversification rate along select lineages correspond to adaptive radiations with movement into the arid zone? To address these questions, we estimated a molecular phylogeny of the Pauropsalta complex using ${\sim}$5300 bp of nucleotide sequence data distributed among five loci (one mtDNA locus and four nDNA loci). We found that this large group of cicadas did not diversify at a constant rate as they spread through Australia; instead the signature of decreasing diversification rate changed roughly around the time of the expansion of the east Antarctic ice sheets ${\sim}$16 Ma and the glaciation of the northern hemisphere ${\sim}$3 Ma. Unlike other Australian taxa, the Pauropsalta complex did not explosively radiate in response to an early invasion of the arid zone. Instead multiple groups invaded the arid zone and experienced rates of diversification similar to mesic-distributed taxa. We found evidence for relictual groups, located in pre-Mesozoic habitat, that have not diversified and continue to reside on mesic hosts in isolated "habitat islands". Future work should focus on groups of similar ages with similar distribution patterns to determine whether this tempo and pattern of diversification and biogeography is consistent with evidence from other phytophagous insects.


Primary care physicians' experiences of carrying out consultations on the internet.
The internet is increasingly used for health matters, including consulting a doctor. Primary care physicians (general practitioners) will probably be involved in performing text-based consultations on the internet as a complement to physical meetings. In the present study, we explored the experiences of GPs already performing consultations on the internet: the challenges, worries and educational demands of the task. A questionnaire was given to 21 GPs performing consultations on the internet for a public, non-commercial 'ask the doctor' service. The questionnaire was carried out at a meeting or sent by mail. The doctors answered a total of 28 questions, 12 of which included graded alternatives. The participating GPs were stimulated and challenged by performing consultations on the internet with previously unknown enquirers, in spite of limitations caused by the lack of personal meetings and physical examinations. The participants experienced a high educational value as a result of the problem-based learning situation induced by unfamiliar questions. The asynchronous feature was appreciated as it allowed time to reflect and perform relevant information searches before replying. Prior training and long-term experience as a family doctor were recommended before embarking on this method of consultation. We conclude that the GPs studied experienced their new role as internet doctors mainly in a positive way, with some limitations. With the increase in consultations on the internet, training in this technique should be integrated into the curricula of medical schools and of continuous professional development (CPD).


Basic Multicolor Flow Cytometry.
Multicolor flow cytometry is a rapidly evolving technology that uses multiple fluorescent markers to identify and characterize cellular subpopulations of interest, allowing rapid analysis on tens of thousands of cells per second, with the possibility of isolating pure, viable populations by cell sorting for further experimentation. This unit covers the tools needed by the beginning immunologist to plan and run multicolor experiments, with information on fluorochromes and their characteristics, spectral spillover, compensation and spread, instrument and reagent variables, and the basic elements of multicolor panel design. Protocols to quantify and maximize sensitivity by titration of reagents and optimization of instrument settings, as well as basic surface and intracellular cell staining, are included. © 2017 by John Wiley & Sons, Inc.


Comparative analysis of mammalian Y chromosomes illuminates ancestral structure and lineage-specific evolution.
Although more than thirty mammalian genomes have been sequenced to draft quality, very few of these include the Y chromosome. This has limited our understanding of the evolutionary dynamics of gene persistence and loss, our ability to identify conserved regulatory elements, as well our knowledge of the extent to which different types of selection act to maintain genes within this unique genomic environment. Here, we present the first MSY (male-specific region of the Y chromosome) sequences from two carnivores, the domestic dog and cat. By combining these with other available MSY data, our multiordinal comparison allows for the first accounting of levels of selection constraining the evolution of eutherian Y chromosomes. Despite gene gain and loss across the phylogeny, we show the eutherian ancestor retained a core set of 17 MSY genes, most being constrained by negative selection for nearly 100 million years. The X-degenerate and ampliconic gene classes are partitioned into distinct chromosomal domains in most mammals, but were radically restructured on the human lineage. We identified multiple conserved noncoding elements that potentially regulate eutherian MSY genes. The acquisition of novel ampliconic gene families was accompanied by signatures of positive selection and has differentially impacted the degeneration and expansion of MSY gene repertoires in different species.


Autoimmune etiology of generalized vitiligo.
Vitiligo is characterized by progressive skin depigmentation resulting from an autoimmune response targeting epidermal melanocytes. Melanocytes are particularly immunogenic by virtue of the contents of their melanosomes, generating the complex radical scavenging molecule melanin in a process that involves melanogenic enzymes and structural components, including tyrosinase, MART-1, gp100, TRP-2 and TRP-1. These molecules are also prime targets of the immune response in both vitiligo and melanoma. The immunogenicity of melanosomal proteins can partly be explained by the dual role of melanosomes, involved both in melanin synthesis and processing of exogenous antigens. Melanocytes are capable of presenting antigens in the context of MHC class II, providing HLA-DR+ melanocytes in perilesional vitiligo skin the option of presenting melanosomal antigens in response to trauma and local inflammation. Type I cytokine-mediated immunity to melanocytes in vitiligo involves T cells reactive with melanosomal antigens, similar to T cells observed in melanoma. In vitiligo, however, T cell tuning allows T cells with higher affinity for melanocyte differentiation antigens to enter the circulation after escaping clonal deletion in primary lymphoid organs. The resulting efficacious and progressive autoimmune response to melanocytes provides a roadmap for melanoma therapy.


Venous thromboembolism: deep venous thrombosis and pulmonary embolism in a neurosurgical population.
Venous thromboembolism (VTE), a combination of deep venous thrombosis (DVT) and pulmonary embolism (PE), is a major cause of morbidity and death in neurosurgical patients. This study evaluates 1) the risk of developing lower-extremity DVT following a neurosurgical procedure; 2) the timing of initiation of pharmacological DVT prophylaxis upon the occurrence of VTE; and 3) the relationship between DVT and PE as related to VTE prophylaxis in neurosurgical patients. The records of all neurosurgical patients between January 2006 and December 2008 (2638 total) were reviewed for clinical documentation of VTE. As part of a quality improvement initiative, a subgroup of 1638 patients was studied during the implementation of pharmacological prophylaxis. A high-risk group of 555 neurosurgical patients in the intensive care unit underwent surveillance venous lower-extremity duplex ultrasonography studies twice weekly. All patients throughout the review received mechanical DVT prophylaxis. Pharmacological DVT prophylaxis, consisting of 5000 U of subcutaneous heparin twice daily (initially started within 48 hours of a neurosurgical procedure and subsequently within 24 hours of a procedure) was implemented in combination with mechanical prophylaxis. The DVT and PE rates were calculated for each group. In the surveillance group (555 patients), 84% of the DVTs occurred within 1 week and 92% within 2 weeks of a neurosurgical procedure. There was a linear correlation between the duration of surgery and DVT development. The use of subcutaneous heparin reduced the rate of DVT from 16% to 9% when medication was given at either 24 or 48 hours postoperatively, without any increase in hemorrhagic complications. In the overall group (2638 patients), there were 94 patients who exhibited clinical signs of a possible PE and therefore underwent spiral CT; 22 of these patients (0.8%) had radiological confirmation of PE. There was no correlation between the use of pharmacological prophylaxis at either time point and the occurrence of PE, despite a 43% reduction in the lower-extremity DVT rate with pharmacological intervention. The majority of DVTs occurred within the first week after a neurosurgical procedure. There was a linear correlation between the duration of surgery and DVT occurrence. Use of early subcutaneous heparin (at either 24 or 48 hours) was associated with a 43% reduction of developing a lower-extremity DVT, without an increase in surgical site hemorrhage. There was no association of pharmacological prophylaxis with overall PE occurrence.


Factors affecting patient and clinician satisfaction with the clinical consultation: can communication skills training for clinicians improve satisfaction?
The provision of adequate information in a clear and sensitive manner can improve cancer patients' experience of care. Satisfaction with the cancer consultation may impact on satisfaction with care in general and adjustment to the disease. This study aims to identify factors that influence patient and clinician satisfaction with the cancer consultation and whether satisfaction can be improved with communication skills training. 160 doctors from 34 UK cancer centres participated. Half were randomized to attend a communication skills training course. Patient satisfaction data are presented at baseline and following a communication skills course or in the case of the control doctors, three months after baseline. Clinicians also rated their satisfaction with the consultations. Overall patient satisfaction was not related to the speciality, seniority or sex of the clinician or patient, site of primary cancer or type of treatment. Satisfaction was related to patients' age, psychological morbidity and, most significantly, satisfaction with the length of wait in clinic. Clinician satisfaction was not related to age, sex or cancer site but clinicians were less satisfied following consultations with patients being treated palliatively. Communication skills training had a non-significant positive effect on patient satisfaction. The subtle benefits of improved communication may be overshadowed by practical problems such as waiting too long to see the doctor, which have an adverse effect on satisfaction.


Formyl peptide receptor 2 expression predicts poor prognosis and promotes invasion and metastasis in epithelial ovarian cancer.
Formyl peptide receptor 2 (FPR2) has been identified as a member of the G protein-coupled chemoattractant receptor (GPCR) family and has been implicated as playing a role in both inflammation and cancer development. Epithelial ovarian cancer (EOC) has been suggested to be correlated with both infectious and non-infectious inflammation. To date, the role of FPR2 in EOC remains poorly understood and controversial. In the present study, we aimed to investigate the potential of FPR2 in regulating EOC. We performed immunohistochemistry and RT-qPCR to analyzed expression of FPR2 in EOC tissues and the correlation between FPR2 and EOC clinicopathological characteristics as well as prognosis were also analyzed. To test the role of FPR2 in EOC cell migration, we established FPR2-knockdown SKOV3 cells and performed wound-healing, Transwell and angiogenesis assays to detect the metastatic potential of these EOC cells. Our studies found that FPR2 was overexpressed in EOC tissues and was positively correlated with EOC clinicopathological characteristics including the International Federation of Gynecology and Obstetrics (FIGO) stage, histological grade and ovarian cancer type. Survival analyses suggested that FPR2 overexpression indicated the poorer prognosis of EOC patients and FPR2 may act as an independent risk factor for EOC prognosis. FPR2 knockdown decreased the migration potential of the ovarian cancer cells. Moreover, serum amyloid A (SAA) may stimulate the migration of SKOV3 cells through FPR2. The present study suggested that FPR2 promoted the invasion and metastasis of EOC and it could be a prognostic marker for EOC.


Program development: role of the clinical nurse specialist in implementing a fast-track postanesthesia care unit.
Advanced practice nurses are involved in many aspects of program development as part of their roles. This can involve such things as developing programs for staff and family education, organizing system-wide quality assurance programs, or implementing new care programs. One unique aspect of the advanced practice nurse's role is the ability to serve as a change agent and implement new models of care. Although all advanced practice nurses can be involved in program development, the role of the Clinical Nurse Specialist lends itself to devoting dedicated services for implementing programmatic change in the clinical setting. This article describes the role of the Clinical Nurse Specialist in implementing an evidence-based, fast-track postanesthesia care unit.


Arrestin/clathrin interaction. Localization of the clathrin binding domain of nonvisual arrestins to the carboxy terminus.
We have recently demonstrated that the nonvisual arrestins, beta-arrestin and arrestin3, interact with high affinity and stoichiometrically with clathrin, and we postulated that this interaction mediates internalization of G protein-coupled receptors (Goodman, O. B., Jr., Krupnick, J. G., Santini, F., Gurevich, V. V., Penn, R. B., Gagnon, A. W., Keen, J. H., and Benovic, J. L. (1996) Nature 383, 447-450). In this study, we localized the clathrin binding domain of arrestin3 using a variety of approaches. Truncation mutagenesis demonstrated that the COOH-terminal half of arrestin3 is required for clathrin interaction. Assessment of the clathrin binding properties of various glutathione S-transferase-arrestin3 fusion proteins indicated that the predominant clathrin binding domain is contained within residues 367-385. Alanine scanning mutagenesis further localized this domain to residues 371-379, and site-directed mutagenesis demonstrated the critical importance of both hydrophobic (Leu-373, Ile-374, and Phe-376) and acidic (Glu-375 and Glu-377) residues in the arrestin3/clathrin interaction. These results are complementary to the observation that hydrophobic and basic residues in clathrin are critical for its interaction with nonvisual arrestins (Goodman, O. B. , Jr., Krupnick, J. G., Gurevich, V. V., Benovic, J. L., and Keen, J. H. (1997) J. Biol. Chem. 272, 15017-15022). Lastly, an arrestin3 mutant in which Leu-373, Ile-374, and Phe-376 were mutated to Ala was significantly defective in its ability to promote beta2-adrenergic receptor internalization in COS-1 cells when compared with wild-type arrestin3. Taken together, these results implicate a discrete region of arrestin3 in high affinity binding to clathrin, an interaction critical for agonist-promoted internalization of the beta2-adrenergic receptor.


Reduced FOXP3(+) regulatory T cells in patients with primary sclerosing cholangitis are associated with IL2RA gene polymorphisms.
Recently, genome wide association studies in primary sclerosing cholangitis (PSC) revealed associations with gene polymorphisms that potentially could affect the function of regulatory T cells (Treg). The aim of this study was to investigate Treg in patients with PSC and to associate their numbers with relevant gene polymorphisms. Treg frequency in blood was assessed by staining for CD4(+)CD25(high)FOXP3(+)CD127(low) lymphocytes and determination of Treg-specific FOXP3 gene locus demethylation. Single nucleotide polymorphisms (SNP) in the interleukin-2 receptor alpha (IL2RA), the interleukin-2 (IL2) and interleukin-21 (IL21) gene locus were analysed. Liver biopsies taken at the time of diagnosis were stained for FOXP3 and CD3. Treg function was assessed in a CFSE-based suppression assay. The frequency of Treg in peripheral blood of PSC patients was significantly decreased. We confirmed this finding by demonstrating a reduction of non-methylated DNA in the Treg-specific demethylated FOXP3 gene region of peripheral blood cells in PSC patients. Reduced peripheral Treg numbers were significantly associated with homozygosity for the major allele of the SNP "rs10905718" in the IL2RA gene. Intrahepatic FOXP3(+) cell numbers at the time of initial diagnosis were decreased in PSC as compared to PBC. In addition to reduced numbers, the suppressive capacity of Treg isolated from PSC patients seemed to be impaired as compared to healthy controls. Our findings indicate that Treg impairment may play a role in the immune dysregulation observed in PSC. Reduced Treg numbers in patients with PSC are associated with polymorphisms in the IL2RA gene.


Contrast-to-noise ratio optimization for a prototype phase-contrast computed tomography scanner.
In the field of biomedical X-ray imaging, novel techniques, such as phase-contrast and dark-field imaging, have the potential to enhance the contrast and provide complementary structural information about a specimen. In this paper, a first prototype of a preclinical X-ray phase-contrast CT scanner based on a Talbot-Lau interferometer is characterized. We present a study of the contrast-to-noise ratios for attenuation and phase-contrast images acquired with the prototype scanner. The shown results are based on a series of projection images and tomographic data sets of a plastic phantom in phase and attenuation-contrast recorded with varying acquisition settings. Subsequently, the signal and noise distribution of different regions in the phantom were determined. We present a novel method for estimation of contrast-to-noise ratios for projection images based on the cylindrical geometry of the phantom. Analytical functions, representing the expected signal in phase and attenuation-contrast for a circular object, are fitted to individual line profiles of the projection data. The free parameter of the fit function is used to estimate the contrast and the goodness of the fit is determined to assess the noise in the respective signal. The results depict the dependence of the contrast-to-noise ratios on the applied source voltages, the number of steps of the phase stepping routine, and the exposure times for an individual step. Moreover, the influence of the number of projection angles on the image quality of CT slices is investigated. Finally, the implications for future imaging purposes with the scanner are discussed.


Associations of hormonal contraceptives and infertility medications on the risk of venous thromboembolism, ischemic stroke, and cardiovascular disease in women.
The purpose of this study was to examine the relations of hormonal contraceptives and infertility drugs with the risk of venous thromboembolism (VTE), deep vein thrombosis (DVT), pulmonary embolism (PE), ischemic stroke, and cardiovascular disease. The Taiwan National Health Institute Research Database was searched for women who had taken hormonal contraceptives or infertility medications from 2000 to 2010. The two groups were age and index date matched with controls (1:4 ratios). Cox regression analysis was used to examine the risks of VTE, DTE, PE, ischemic stroke, and cardiovascular disease. A total of 32,067 women were included in the hormonal contraceptives group and 4710 in the infertility medications group (matched controls: 127,872 and 18,840, respectively). After adjustment for age, comorbidities, and other confounders, the contraceptives group had a higher risk of VTE (adjusted HR 1.14, 95% CI 1.004 to 1.30) and cardiovascular disease (adjusted HR 1.30, 95% CI 1.26 to 1.34), and lower risk of ischemic stroke (adjusted HR 0.90, 95% CI 0.86 to 0.95). The infertility medications group had a higher risk of VTE (adjusted HR 1.996, 95% CI 1.41 to 2.72) and DVT (adjusted HR 1.86, 95% CI 1.31 to 2.63), and lower risk of ischemic stroke (adjusted HR 0.82, 95% CI 0.68 to 0.99) and cardiovascular disease (adjusted HR 0.83, 95% CI 0.74 to 0.94). Hormonal contraceptives and infertility medications appear to lower the risk of ischemic stroke and increase the risk of VTE; however, their effect on the risk of other types of cardiovascular events varies.


Structure-function relationships in immobilized chymotrypsin catalysis.
Specific activities and the amounts of active immobilized enzyme were determined for several different preparations of alpha-chymotrypsin immobilized on CNBr-activated Sepharose 4B. Electron paramagnetic resonance (EPR) spectroscopy of free and immobilized enzyme with a spin label coupled to the active site was used to probe the effects of different immobilization conditions on the immobilized enzyme active site configuration. Specific activity of active enzyme decreased and rotational correlation time of the spin label increased with increasing immobilized enzyme loading. Enzyme immobilized using an intermediate six-carbon spacer arm exhibited greater specific activity and spin label mobility than directly coupled enzyme. The observed activity changes due to immobilization were completely consistent with corresponding active site structure alterations revealed by EPR spectroscopy.


CCN6 (WISP3) decreases ZEB1-mediated EMT and invasion by attenuation of IGF-1 receptor signaling in breast cancer.
During progression of breast cancer, CCN6 protein exerts tumor inhibitory functions. CCN6 is a secreted protein that modulates the insulin-like growth factor-1 (IGF-1) signaling pathway. Knockdown of CCN6 in benign mammary epithelial cells triggers an epithelial to mesenchymal transition (EMT), with upregulation of the transcription factor ZEB1/δEF1. How CCN6 regulates ZEB1 expression is unknown. We hypothesized that CCN6 might regulate ZEB1, EMT and breast cancer invasion by modulating IGF-1 signaling. Exogenously added human recombinant CCN6 protein was sufficient to downregulate ZEB1 mRNA and protein levels in CCN6-deficient (CCN6 KD) HME cells and MDA-MB-231 breast cancer cells. Recombinant CCN6 protein decreased invasion of CCN6 KD cells compared with controls. We discovered that knockdown of CCN6 induced IGF-1 secretion in HME cells cultivated in serum-free medium to higher concentrations than found in MDA-MB-231 cells. Treatment with recombinant CCN6 protein was sufficient to decrease IGF-1 protein and mRNA to control levels, rescuing the effect of CCN6 knockdown. Specific inhibition of IGF-1 receptors using the pharmacological inhibitor NVP-AE541 or short hairpin shRNAs revealed that ZEB1 upregulation due to knockdown of CCN6 requires activation of IGF-1 receptor signaling. Recombinant CCN6 blunted IGF-1-induced ZEB1 upregulation in MDA-MB-231 cells. Our data define a pathway in which CCN6 attenuates IGF-1 signaling to decrease ZEB1 expression and invasion in breast cancer. These results suggest that CCN6 could be a target to prevent or halt breast cancer invasion.


[Aerococcus urinae endocarditis: first case report in Belgium and review of the literature].
Aerococcus urinae is a rarely reported pathogen, possibly due to difficulties in the identification of the organism. Only sixteen cases of A. urinae endocarditis have yet been described. Generally, patients were males with predisposing conditions and the outcomes were bad. We describe the first Belgian case of endocarditis due to A. urinae. Which occurred to a 79-year-old female patient. The diagnostic of A. urinae endocarditis was based on the cardiac ultrasonography and the isolation of the bacteria in 6/7 pairs of blood culture. The patient has been successfully treated by a long course of intravenous antibiotics (38 days) and surgery (aortic valve replacement by bioprothesis). We review the 14 cases published in the English literature and discuss the best clinical management of this type of endocarditis.


Equilibrium properties of warm dense deuterium calculated by the wave packet molecular dynamics and density functional theory method.
A joint simulation method based on the wave packet molecular dynamics and density functional theory (WPMD-DFT) is applied to study warm dense deuterium (nonideal deuterium plasmas). This method was developed recently as an extension of the wave packet molecular dynamics (WPMD) in which the equations of motion are solved simultaneously for classical ions and semiclassical electrons represented as Gaussian wave packets. Compared to the classical molecular dynamics and WPMD simulations, the method of WPMD-DFT provides a more accurate representation of quantum effects such as electron-ion coupling and electron degeneracy. It allows studying nonadiabatic dynamics of electrons and ions in equilibrium and nonequilibrium states while being more accurate and efficient at high densities than WPMD and classical molecular dynamics. In the paper, we discuss particular features of the method such as special boundary conditions and the procedure of isentrope calculation as well as the results obtained by WPMD-DFT for the shock-compressed deuterium. The compression isentrope and principal Hugoniot curves obtained by WPMD-DFT are compared with available experimental data and other simulation approaches to validate the method. It opens up a possibility of further application of the method to study nonequilibrium states and relaxation processes.


G-OnRamp: Generating genome browsers to facilitate undergraduate-driven collaborative genome annotation.
Scientists are sequencing new genomes at an increasing rate with the goal of associating genome contents with phenotypic traits. After a new genome is sequenced and assembled, structural gene annotation is often the first step in analysis. Despite advances in computational gene prediction algorithms, most eukaryotic genomes still benefit from manual gene annotation. This requires access to good genome browsers to enable annotators to visualize and evaluate multiple lines of evidence (e.g., sequence similarity, RNA sequencing [RNA-Seq] results, gene predictions, repeats) and necessitates many volunteers to participate in the work. To address the technical barriers to creating genome browsers, the Genomics Education Partnership (GEP; https://gep.wustl.edu/) has partnered with the Galaxy Project (https://galaxyproject.org) to develop G-OnRamp (http://g-onramp.org), a web-based platform for creating UCSC Genome Browser Assembly Hubs and JBrowse genome browsers. G-OnRamp also converts a JBrowse instance into an Apollo instance for collaborative genome annotations in research and educational settings. The genome browsers produced can be transferred to the CyVerse Data Store for long-term access. G-OnRamp enables researchers to easily visualize their experimental results, educators to create Course-based Undergraduate Research Experiences (CUREs) centered on genome annotation, and students to participate in genomics research. In the process, students learn about genes/genomes and about how to utilize large datasets. Development of G-OnRamp was guided by extensive user feedback. Sixty-five researchers/educators from >40 institutions participated through in-person workshops, which produced >20 genome browsers now available for research and education. Genome browsers generated for four parasitoid wasp species have been used in a CURE engaging students at 15 colleges and universities. Our assessment results in the classroom demonstrate that the genome browsers produced by G-OnRamp are effective tools for engaging undergraduates in research and in enabling their contributions to the scientific literature in genomics. Expansion of such genomics research/education partnerships will be beneficial to researchers, faculty, and students alike.


Comparison of the phytochemical profiles of five native plant species in two different forest formations.
The expression of chemical compounds by individual plants of the same species in different locations may be affected by abiotic factors resulting in differences in the production of allelopathic compounds. The objective of this study was to compare the phytochemical profiles of plant species from two different forest formations in the state of Paraná, Brazil. The forest formations were Seasonal Semideciduous Forest (SSF) and Lowland Ombrophilous Dense Forest (LODF), and the five study species were Jacaranda micrantha, Cecropia pachystachya, Mimosa bimucronata, Schinus terebinthifolius and Cedrela fissilis. Secondary metabolites were extracted by exhaustive extraction with methanol, and the crude extract was fractionated using column chromatography. The fractions were used to calculate the retention factor of the main compounds using thin layer chromatography and phytochemical tests. The classes of compounds identified were practically the same among the analyzed species, however, at different levels of concentration. The type of tannins found in S. terebinthifolius differed between the two forest formations.


Stabilization of Foxp3 by Targeting JAK2 Enhances Efficacy of CD8 Induced Regulatory T Cells in the Prevention of Graft-versus-Host Disease.
CD8+ induced regulatory T cells (iTregs) have been identified to suppress alloreactive immune responses and expressed regulatory T cell (Treg) ontological markers as similar as CD4+ iTregs. However, adoptive transfer of CD8+ iTreg-based therapy is hampered by the instability of Treg specific-transcription factor, Foxp3. As CD8+ iTregs were previously demonstrated to possess superior tumor-killing ability to CD4+ iTregs, adoptive transfer of stabilized CD8+ iTregs would be a potential therapy to prevent tumor relapse during graft-versus-leukemia disease (GVHD) treatment. In the current study, we generated alloantigen reactive CD8+ iTregs from JAK2-/- T cells and adoptively transferred them to MHC-mismatched and haploidentical murine models of allogeneic bone marrow transplantation. JAK2-/- CD8+ iTregs not only attenuated GVHD but also preserved graft-versus-leukemia effect. Mechanistic analysis revealed that JAK2-/- CD8+ iTregs upregulated natural Treg marker (neuropilin-1), and augmented DNA demethylation of CNS2 region within Foxp3 gene. These properties licensed JAK2-/- CD8+ iTregs to retain high Foxp3 expression resulting in less conversion to type 1 CTLs; as a result, JAK2-/- CD8+ iTregs were able to maintain their suppressive and cytolytic function. Thus, our findings provide a strong rationale and means to stabilize CD8+ iTregs by targeting JAK2, and the stabilized CD8+ iTregs exhibit therapeutic potential for alleviating GVHD and preserving the graft-versus-leukemia effect.


Enhanced expression of p53 and apoptosis induced by blockade of the vacuolar proton ATPase in cardiomyocytes.
Activation of the vacuolar proton ATPase (VPATPase) has been implicated in the prevention of apoptosis in neutrophils and adult cardiac myocytes. To determine the role of the VPATPase in apoptosis of cardiac myocytes, we used a potent and specific inhibitor of the VPATPase, bafilomycin A1. Bafilomycin A1 alone caused increased DNA laddering of genomic DNA and increased nuclear staining for fragmented DNA in neonatal cardiomyocyte apoptosis in a dose- and time-dependent manner. Intracellular acidification in cardiac myocytes was also observed after 18 h of bafilomycin A1 treatment. Accordingly, bafilomycin A1-treated myocytes also showed increased accumulation of p53 protein and p53-dependent transactivation of gene expression, including a persistent upregulation of p21/wild-type p53 activated fragment 1/cyclin kinase inhibitor protein-1 mRNA. The bafilomycin A1-induced increase in p53 protein levels was accompanied by a marked increase in p53 mRNA accumulation. In contrast, cardiac fibroblasts treated with bafilomycin A1 showed no change in p53 protein expression or pHi and did not undergo apoptosis even after 24 h of treatment. Our data suggest that blockade of the VPATPase induces apoptotic cell death of cardiac myocytes and that this may occur through a p53-mediated apoptotic pathway.


The economics of peritoneal dialysis: a cost-efficiency study.
To determine the maximum possible return of urea extracted per litre of peritoneal dialysis fluid used, nine test exchanges using volumes of 1, 1.5, and 2 litres of fluid for periods of 0.5, 1, and 1.5 h, were studied in 15 consecutive peritoneal dialyses. The actual quantity of urea extracted was estimated. On comparison, it was found that the best return was achieved with 1-litre volumes used for 1.5 or 1 h (80.65 +/- 5.58 and 80.18 +/- 5.13% respectively of a possible best of 100%) the difference between these being insignificant. All other exchange volumes and times used yielded significantly worse results. We recommend the use of 1-hour, 1-litre exchanges in peritoneal dialysis, except in patients who are in a hypercatabolic state, in whom we would use haemodialysis or 0.5 h 2-litre exchanges of peritoneal dialysis.


Multihit two-dimensional charged-particle imaging system with real-time image processing at 1000 frames/s.
A high-speed imaging system developed for two-dimensional counting of charged particles is presented. Microchannel plates coupled with a phosphor screen of a short emission lifetime (<1 micros) are used to visualize the two-dimensional positions of charged-particle impacts, and the image on the phosphor screen is captured with a 1 kHz complementary metal oxide semiconductor (CMOS) image sensor (512x512 pixels). A multistage image intensifier consisting of the first and second generation devices was used to compensate for the low sensitivity of CMOS. The centers of gravity (COG) of individual light spots in each image frame are calculated in real time by a field programmable gate array circuit. The performance of this system is tested by time-resolved photoelectron imaging (TR-PEI) of NO using (1+1(')) resonance enhanced multiphoton ionization via the A (2)Sigma(+) state with a femtosecond laser operated at 1 kHz. The new system enabled COG detection for more than ten particles in each frame at 1 kHz and achieved an extremely high degree of accuracy in the measurement of photoelectron angular distributions in TR-PEI.


A novel fluorescence method for activity assay and drug screening of T4 PNK by coupling rGO with ligase reaction.
T4 polynucleotide kinase (PNK) is the primary member of the 5'-kinase family that can transfer the γ-phosphate residue of ATP to the 5'-hydroxyl group of oligonucleotides. In this article, using the differential quenching ability of reduced graphene oxide (rGO) towards the fluorophore-labeled DNA probe, we propose a novel method for detecting T4 PNK activity assisted by ligase reaction. Under the optimized conditions, the detection limit of T4 PNK was estimated to be 0.0002 U μL-1 in the linear region of 0.001 U μL-1-0.1 U μL-1. Additionally, the developed method was used to screen regulators of T4 PNK from natural compounds. The compound f isolated from the root of Kadsura coccinea (Lem.) A.C. Smith was found to stimulate T4 PNK activity in a concentration-dependent manner in vitro. Finally, the method was used to monitor the relation of T4 PNK activity with pelvic inflammatory disease (PID). The results demonstrated that the development of this disease could inhibit T4 PNK activity to some extent. In summary, the above data indicate that the method not only provides a universal platform for monitoring T4 PNK activity, but also shows great potential to be used in drug screening and clinic diagnosis.


The TLR2/TLR6 ligand FSL-1 mitigates radiation-induced hematopoietic injury in mice and nonhuman primates.
Thrombocytopenia, hemorrhage, anemia, and infection are life-threatening issues following accidental or intentional radiation exposure. Since few therapeutics are available, safe and efficacious small molecules to mitigate radiation-induced injury need to be developed. Our previous study showed the synthetic TLR2/TLR6 ligand fibroblast stimulating lipopeptide (FSL-1) prolonged survival and provided MyD88-dependent mitigation of hematopoietic acute radiation syndrome (H-ARS) in mice. Although mice and humans differ in TLR number, expression, and function, nonhuman primate (NHP) TLRs are like those of humans; therefore, studying both animal models is critical for drug development. The objectives of this study were to determine the efficacy of FSL-1 on hematopoietic recovery in small and large animal models subjected to sublethal total body irradiation and investigate its mechanism of action. In mice, we demonstrate a lack of adverse effects, an easy route of delivery (subcutaneous) and efficacy in promoting hematopoietic progenitor cell proliferation by FSL-1. NHP given radiation, followed a day later with a single subcutaneous administration of FSL-1, displayed no adversity but showed elevated hematopoietic cells. Our analyses revealed that FSL-1 promoted red blood cell development and induced soluble effectors following radiation exposure. Cytologic analysis of bone marrow aspirates revealed a striking enhancement of mononuclear progenitor cells in FSL-1-treated NHP. Combining the efficacy of FSL-1 in promoting hematopoietic cell recovery with the lack of adverse effects induced by a single administration supports the application of FSL-1 as a viable countermeasure against H-ARS.


PNPLA1 defects in patients with autosomal recessive congenital ichthyosis and KO mice sustain PNPLA1 irreplaceable function in epidermal omega-O-acylceramide synthesis and skin permeability barrier.
Autosomal recessive congenital ichthyosis (ARCI) is a heterogeneous group of monogenic genodermatoses that encompasses non-syndromic disorders of keratinization. The pathophysiology of ARCI has been linked to a disturbance in epidermal lipid metabolism that impaired the stratum corneum function, leading to permeability barrier defects. Functional characterization of some genes involved in ARCI contributed to the identification of molecular actors involved in epidermal lipid synthesis, transport or processing. Recently, PNPLA1 has been identified as a gene causing ARCI. While other members of PNPLA family are key elements in lipid metabolism, the function of PNPLA1 remained unclear. We identified 5 novel PNPLA1 mutations in ARCI patients, mainly localized in the putative active enzymatic domain of PNPLA1. To investigate Pnpla1 biological role, we analysed Pnpla1-deficient mice. KO mice died soon after birth from severe epidermal permeability defects. Pnpla1-deficient skin presented an important impairment in the composition and organization of the epidermal lipids. Quantification of epidermal ceramide species highlighted a blockade in the production of ω-O-acylceramides with a concomitant accumulation of their precursors in the KO. The virtually loss of ω-O-acylceramides in the stratum corneum was linked to a defective lipid coverage of the resistant pericellular shell encapsulating corneocytes, the so-called cornified envelope, and most probably disorganized the extracellular lipid matrix. Finally, these defects in ω-O-acylceramides synthesis and cornified envelope formation were also evidenced in the stratum corneum from PNPLA1-mutated patients. Overall, our data support that PNPLA1/Pnpla1 is a key player in the formation of ω-O-acylceramide, a crucial process for the epidermal permeability barrier function.


Analysis of the cost of care for children and adolescents with medical complex chronic conditions.
This paper aimed to identify the use of technology and to analyze the cost of hospital care for children and adolescents with medical complex chronic conditions at a public federal hospital specialized in high-complexity pediatric care, and was performed concomitantly with a prospective cohort study conducted over a one-year period. It included 146 patients with complex medical chronic conditions and 37 non-chronic patients. The analysis showed that most patients had, on average, two hospitalizations a year and were diagnosed with diseases related to at least two organic systems. Catheters, drains and gastrostomy were the most common technologies used. Median direct costs of patients with medically complex chronic conditions were higher than those of non-chronic patients when comparing the use of technology. The study shows high hospitalization cost to these patients. Technology use and hospitalization care costs documentation yields more data to support decision-makers in the planning, managing, and financing of pediatric health policies.


Monoclonal antibodies reveal novel aspects of the biochemistry and organization of olfactory neurons following unilateral olfactory bulbectomy.
Following unilateral olfactory bulbectomy in rats the ipsilateral olfactory neuroepithelium undergoes degeneration. Subsequently, the receptor neuron complement of the tissue is restored by the proliferation and differentiation of immature neuroblasts. However, as noted by other workers, in the absence of a target organ the dynamics of neuron regeneration is altered such that there is an overall reduction in the number of cells positive for the olfactory marker protein when cellular equilibrium is re-established. Immunocytochemical staining of the olfactory epithelium of unilaterally bulbectomized rats with a series of anti-neuronal monoclonal antibodies reveals an attenuation of binding of some antibodies to the neurons of the ipsilateral epithelium. In contrast, other anti-neuronal monoclonal antibodies show no difference in staining intensity when ipsilateral and control contralateral epithelia are compared. These data suggest that the expression of some neuronal antigens is subject to control by the target olfactory bulb, whereas others are independent of such putative regulation. Besides altering the expression of some antigenic determinants, olfactory bulbectomy also results in certain organizational changes in epithelium. First, bulbectomy produces an increase in the incidence of a cell type that appears to span the neuroepithelium. Although the morphology of these cells is more akin to a sustentacular cell than to a receptor neuron, they are not immunoreactive with antibodies to sustentacular cells. The cells are stained, however, by an anti-neuronal antibody, NEU-9. The second aspect of altered organization is the appearance of novel olfactory marker protein-positive structures in the olfactory mucosa.(ABSTRACT TRUNCATED AT 250 WORDS)


Laser inactivation protein patterning of cell culture microenvironments.
Protein micropatterned substrates have emerged as important tools for studying how cells interact with their environment, as well as allowing useful experimental control over, for example, cell shape and cell position on a surface. Here we present a new approach for protein micropatterning in which a focused laser is used to locally inactivate proteins on a protein-coated substrate. By translating the laser relative to the substrate, protein patterns of essentially arbitrary shape can be produced. This approach has a number of useful features. To begin, it is a maskless writing approach. Thus new patterns can be designed and implemented quickly. Laser inactivation can also be performed on a number of different substrate materials, ranging from glass to polydimethylsiloxane. Further, the inactivation is dose dependent, thus complex gradients and other non-uniform distributions of proteins can be produced. Because the focus of the laser can be changed quickly, laser-based patterning can also be applied to substrates with complex topographies or enclosed surfaces--as long as an optical path is available. To demonstrate this capability, protein patterns were made on the inside of small quartz capillary tubes. Patterned substrates produced using laser inactivation constrain cell shape in predictable ways, and we show that these substrates are compatible with a number of different eukaryotic cell lines.


Dynamic nuclear polarization of (1)H, (13)C, and (59)Co in a tris(ethylenediamine)cobalt(III) crystalline lattice doped with Cr(III).
The study of inorganic crystalline materials by solid-state NMR spectroscopy is often complicated by the low sensitivity of heavy nuclei. However, these materials often contain or can be prepared with paramagnetic dopants without significantly affecting the structure of the crystalline host. Dynamic nuclear polarization (DNP) is generally capable of enhancing NMR signals by transferring the magnetization of unpaired electrons to the nuclei. Therefore, the NMR sensitivity in these paramagnetically doped crystals might be increased by DNP. In this paper we demonstrate the possibility of efficient DNP transfer in polycrystalline samples of [Co(en)3Cl3]2·NaCl·6H2O (en = ethylenediamine, C2H8N2) doped with Cr(III) in varying concentrations between 0.1 and 3 mol %. We demonstrate that (1)H, (13)C, and (59)Co can be polarized by irradiation of Cr(III) with 140 GHz microwaves at a magnetic field of 5 T. We further explain our findings on the basis of electron paramagnetic resonance spectroscopy of the Cr(III) site and analysis of its temperature-dependent zero-field splitting, as well as the dependence of the DNP enhancement factor on the external magnetic field and microwave power. This first demonstration of DNP transfer from one paramagnetic metal ion to its diamagnetic host metal ion will pave the way for future applications of DNP in paramagnetically doped materials or metalloproteins.


On the Operational Aspects of Measuring Nanoparticle Sizes.
Nanoparticles are defined as elementary particles with a size between 1 and 100 nm for at least 50% (in number). They can be made from natural materials, or manufactured. Due to their small sizes, novel toxicological issues are raised and thus determining the accurate size of these nanoparticles is a major challenge. In this study, we performed an intercomparison experiment with the goal to measure sizes of several nanoparticles, in a first step, calibrated beads and monodispersed SiO₂ Ludox®, and, in a second step, nanoparticles (NPs) of toxicological interest, such as Silver NM-300 K and PVP-coated Ag NPs, Titanium dioxide A12, P25(Degussa), and E171(A), using commonly available laboratory techniques such as transmission electron microscopy, scanning electron microscopy, small-angle X-ray scattering, dynamic light scattering, wet scanning transmission electron microscopy (and its dry state, STEM) and atomic force microscopy. With monomodal distributed NPs (polystyrene beads and SiO₂ Ludox®), all tested techniques provide a global size value amplitude within 25% from each other, whereas on multimodal distributed NPs (Ag and TiO₂) the inter-technique variation in size values reaches 300%. Our results highlight several pitfalls of NP size measurements such as operational aspects, which are unexpected consequences in the choice of experimental protocols. It reinforces the idea that averaging the NP size from different biophysical techniques (and experimental protocols) is more robust than focusing on repetitions of a single technique. Besides, when characterizing a heterogeneous NP in size, a size distribution is more informative than a simple average value. This work emphasizes the need for nanotoxicologists (and regulatory agencies) to test a large panel of different techniques before making a choice for the most appropriate technique(s)/protocol(s) to characterize a peculiar NP.


Nurse-led postdischarge telephone follow-up calls: A mixed study systematic review.
To explore the quantitative and qualitative literature on the impact of nurse-led postdischarge telephone follow-up (TFU) call interventions on patient outcomes. Adverse patient outcomes such as postdischarge problems, premature contact with health systems, inability to self-manage conditions and hospital readmissions all have an impact on the health and well-being, and satisfaction of patients as well as a financial impact on healthcare systems. A mixed-study systematic review. A systematic search of CINAHL, Ebsco, PubMed, Quest and Cinch-Health databases was undertaken using the key terms "nurs*," "nurse-led," "nurse initiated," "discharge," "hospital," "telephone," "follow-up" and "telephone follow-up" to identify relevant original peer-reviewed studies published between 2010-2016. Ten articles were selected for inclusion. The selected papers were critically appraised. A sequential explanatory approach with a convergent synthesis was used to report findings following PRISMA guidelines. The findings demonstrate that nurse-led TFU interventions have the potential to improve patient outcomes. The studies suggest patient satisfaction with TFU is one of the strongest positive outcomes from the interventions. However, the results do not support improvement in patient readmission or mortality. Of the 10 studies reviewed, only two were methodologically strong limiting the conclusions that can be drawn from the current research on this topic. Telephone follow-up interventions improve patient satisfaction and have the potential to meet patient information and communication needs, improve self-management and follow-up appointment attendance and reduce postdischarge problems. Further research is required to explore patients' perceptions of the most useful content of TFU calls, the efficacy of TFU calls and nurses' perceptions and experiences of conducting TFU interventions. When conducted by a nurse, these interventions have the potential to enhance postdischarge care to patients and meet care needs. Patients perceive TFU as acceptable and are satisfied with this form of postdischarge care.


Aspects of mycobacterial response to beryllate ions in vitro.
Comparative growth measurements of the mycobacterial species Mycobacterium phlei and Mycobacterium tuberculosis were carried out with liquid cultures supplemented with oxalatoberyllate ions in the concentration range 0-120 microM Be. For both species, the effect of beryllium on specific growth parameters can be represented by a Be concentration-dependent, exponential expression. Evidence for beryllium binding to cellular phospholipids (approximately 15% of Be uptake) is provided by 31P NMR spectroscopy, thin-layer chromatography and atomic absorption spectrometry.


Systematic review: the treatment of anal fissure.
Anal fissure is one of the most common anorectal conditions encountered in clinical practice. Most patients experience anal pain with defecation and minor bright red rectal bleeding, allowing a focused history to direct the evaluation. A systematic medical literature search of NIH, Pubmed, and MEDLINE using the search terms anal fissure, sphincterotomy, anal surgery and anal fissure medical therapy. English language was not a restriction. Cited references were used to find additional studies. No single treatment is the best choice for all patients. Because pharmacological therapy is not associated with permanent alterations in continence, a trial of either a topical sphincter relaxant or botulin toxin injection, along with adequate fluid and fibre intake, is a reasonable option. However, because pharmacological therapy has lower healing and higher relapse rates, surgery can be offered in the first instance to patients without incontinence risk factors who have severe, unrelenting pain and are willing to accept a small risk of incontinence, for the highest likelihood of prompt healing and the lowest risk of recurrence. Both non-operative and operative approaches currently exist for the management of anal fissure. Improved non-surgical therapies may continue to lessen the role of sphincter-dividing surgery in future.


Photoperiodic regulation of leptin sensitivity in the Siberian hamster, Phodopus sungorus, is reflected in arcuate nucleus SOCS-3 (suppressor of cytokine signaling) gene expression.
We present the first evidence that suppressor of cytokine signaling-3 (SOCS3), a protein inhibiting Janus kinase/signal transducer and activator of transcription (STAT) signaling distal of the leptin receptor, conveys seasonal changes in leptin sensitivity in the Siberian hamster. Food deprivation (48 h) reduced SOCS3 gene expression in hamsters acclimated to either long (LD) or short (SD) photoperiods, suggesting that leptin signals acute starvation regardless of photoperiod. However, SOCS3 mRNA levels were substantially lower in the hypothalamic arcuate nucleus of hamsters acclimated to SD than in those raised in LD. In juveniles raised in LD, a rapid increase in SOCS3 mRNA was observed within 4 d of weaning, which was completely prevented by transfer to SD on the day of weaning. The early increase in SOCS3 gene expression in juvenile hamsters in LD clearly preceded the establishment of different body weight trajectories in LD and SD. In adult LD hamsters, SOCS3 mRNA was maintained at an elevated level despite the chronic food restriction imposed to lower body weight and serum leptin to or even below SD levels. A single injection of leptin in SD hamsters elevated SOCS3 mRNA to LD levels, whereas leptin treatment had no effect on SOCS3 gene expression in LD hamsters. Our results suggest that the development of leptin resistance in LD-acclimated hamsters involves SOCS3-mediated suppression of leptin signaling in the arcuate nucleus. Increased SOCS3 expression in LD hamsters is independent of body fat and serum leptin levels, suggesting that the photoperiod is able to trigger the biannual reversible switch in leptin sensitivity.


Algorithm for the treatment of schizophrenia in Japan.
Evidence-based psychopharmacological algorithms for the treatment of patients with schizophrenia have been developed in many countries in the last decade. While it would be of interest to consider a common algorithm based on international consensus, algorithms and information on antipsychotics available in each country are limited. Inspired by the algorithm generated by the International Psychopharmacology Algorithm (IPA) Project, this algorithm for the treatment of schizophrenia has been developed by the Japan Psychophamacology Algorithm (JPA) Project. New antipsychotics, such as clozapine, olanzapine and quetiapine, are excluded from this algorithm, being currently unavailable in Japan. In the end there was no essential difference between the algorithms for the treatment of acute schizophrenic episodes. However, combined use of antipsychotics appears to be more common in Japan and the adjunctive use of L-DOPS or thyrotropin-releasing hormone is included in the JPA algorithm for the treatment of drug-refractory schizophrenia.