Identification of surface-exposed outer membrane antigens of Helicobacter pylori.
Despite the potential significance of surface-localized antigens in the colonization by and disease processes of Helicobacter pylori, few such components have been unequivocally identified and/or characterized. To further investigate the surface of this bacterium, monoclonal antibodies (MAbs) to a sarcosine-insoluble outer membrane fraction prepared from H. pylori NCTC 11637 were raised. MAbs were selected on the basis of their surface reactivity to whole cells by enzyme-linked immunosorbent assay, immunofluorescence, and immunoelectron microscopy. By use of this selection protocol, 14 surface-reactive MAbs were chosen. These MAbs were used to identify six protein antigens (molecular masses, 80, 60, 51, 50, 48, and 31 kDa), all of which were localized within or associated with the outer membrane. Two of the MAbs recognized the core region of lipopolysaccharide (LPS). Only these two anti-LPS MAbs also recognized the flagellar sheath, indicating a structural difference between the sheath and outer membrane. Three of the protein antigens (80, 60, and 51 kDa) were strain specific, while the other three antigens were present in other strains of H. pylori. Both the 51- and 48-kDa antigens were heat modifiable and likely are porins. A conserved 31-kDa protein may represent another species of porin. A method involving sucrose density ultracentrifugation and Triton extraction that allows the preparation of H. pylori outer membranes with minimal inner membrane contamination is described. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed that the protein content of the H. pylori outer membrane is similar structurally to those of other species of Helicobacter but markedly different from those of taxonomically related Campylobacter spp. and Escherichia coli. H. pylori also appeared to lack peptidoglycan-associated proteins.


Differential Effects of Cold Atmospheric Plasma in the Treatment of Malignant Glioma.
Cold atmospheric plasma (CAP) has recently been shown to selectively target cancer cells with minimal effects on normal cells. We systematically assessed the effects of CAP in the treatment of glioblastoma. Three glioma cell lines, normal astrocytes, and endothelial cell lines were treated with CAP. The effects of CAP were then characterized for viability, cytotoxicity/apoptosis, and cell cycle effects. Statistical significance was determined with student's t-test. CAP treatment decreases viability of glioma cells in a dose dependent manner, with the ID50 between 90-120 seconds for all glioma cell lines. Treatment with CAP for more than 120 seconds resulted in viability less than 35% at 24-hours posttreatment, with a steady decline to less than 20% at 72-hours. In contrast, the effect of CAP on the viability of NHA and HUVEC was minimal, and importantly not significant at 90 to 120 seconds, with up to 85% of the cells remained viable at 72-hours post-treatment. CAP treatment produces both cytotoxic and apoptotic effects with some variability between cell lines. CAP treatment resulted in a G2/M-phase cell cycle pause in all three cell lines. This preliminary study determined a multi-focal effect of CAP on glioma cells in vitro, which was not observed in the non-tumor cell lines. The decreased viability depended on the treatment duration and cell line, but overall was explained by the induction of cytotoxicity, apoptosis, and G2/M pause. Future studies will aim at further characterization with more complex pre-clinical models.


Move it or Lose it. Is it Reasonable for Older Adults with Osteoarthritis to Continue to Use Paracetamol in Order to Maintain Physical Activity?
Osteoarthritis is a common progressive disease in older adults, and those affected often have impaired physical function, co-existing disease states, and reduced quality of life. In patients with osteoarthritis, pain is reported as a primary cause of mobility limitation, and guidelines recommend a mix of pharmacologic and non-pharmacologic strategies for pain management. The benefits of exercise in the management of osteoarthritis are well established; however, pain appears to be the biggest barrier to patients engaging in, and adhering to, physical activity programs. Attitudes towards the use of pain medications differ widely, and lack of efficacy or fear of side effects may lead to sub-therapeutic dosing. Furthermore, a recent review suggesting that short-term paracetamol use is ineffective for osteoarthritis has added to the confusion. This narrative review investigates limitations of current medications, summarizes patient attitudes toward the use of analgesics for osteoarthritis pain (with a focus on paracetamol), and explores the uptake of physical activity for osteoarthritis management. Evidence suggests that, despite clear guidelines, symptoms of osteoarthritis generally remain poorly managed. More research is required to investigate clinical outcomes in patients with osteoarthritis through optimized medication plans to better understand whether longer-term analgesic use in conjunction with physical activity can assist patients to overcome mobility limitations.


Reduced glutamate immunolabeling in the nucleus accumbens following extended withdrawal from self-administered cocaine.
Alterations in the density of GABA and glutamate immunolabeling within nerve terminals in the shell region of the nucleus accumbens were assessed in rats withdrawn from intravenous cocaine exposure. Four groups of rats were used: one group self-administered cocaine (0.42 mg/kg/infusion) in daily 3-h sessions for approximately 2 weeks, two additional groups received either saline or cocaine in a noncontingent fashion, and a fourth comprised a drug-naive, age-matched control group. Immunogold electron microscopy was used to quantify presynaptic terminal GABA and glutamate density within the vesicular and mitochondrial pools approximately 18 days following the last drug or saline exposure in the treatment groups. A significant 27.7% decrease in vesicular glutamate density within asymmetrical nerve terminals was observed in animals that self-administered cocaine as compared to controls. This group also showed an 18.6% decrease in vesicular nerve terminal glutamate immunolabeling as compared to animals that were administered a similar total dose of cocaine in a response-independent fashion. No significant changes in the density of nerve terminal GABA vesicular immunolabeling were observed in any groups. For both transmitters, no differences were detected in the density of immunolabeling within the presynaptic mitochondrial (i.e., metabolic) pool. These results demonstrate that glutamate density is suppressed in the shell region of the nucleus accumbens following withdrawal from 2 weeks of cocaine exposure. The findings also suggest that the motivational aspects that accompany self-administration may participate in this reduction.


Identification of rabbit cytochromes P450 2C1 and 2C2 as arachidonic acid epoxygenases.
Microsomes prepared from COS-1 cells transiently expressing rabbit cytochromes P450 2C1 and 2C2 catalyzed the metabolism of arachidonic acid to predominantly 11,12- and 14,15-epoxyeicosatrienoic acids (EETs) when microsomal epoxide hydrolase activity was inhibited by 0.2 mM 1,2-epoxy-3,3,3-trichloropropane. P450 2C2 catalyzed the formation of 11,12-EET and 14,15-EET at a ratio of 3.0 and also produced 19-hydroxyeicosatetraenoic acid (19-HETE). The 11,12-EET, 14,15-EET, and 19-HETE represented 48.3, 15.9, and 12.8%, respectively, of the total metabolites formed. P450 2C1 produced a similar but distinct ratio of 11,12-EET to 14,15-EET (2.0) and did not produce any detectable 19-HETE. The 11,12-EET and 14,15-EET represented 63.0 and 31.1%, respectively, of the total metabolites formed. The 8,9- and 5,6-EETs were not detected with either enzyme. The ratio of the 11,12-EET to 14,15-EET was 1.5 with P450 2CAA, a P450 arachidonic acid epoxygenase (P450 2CAA) that had an amino-terminal sequence identical to that of P450 2C2 [J. Biol. Chem. 267:5552-5559 (1992)]. P450 2C1, 2C2, and 2CAA metabolized lauric acid. The ratio of omega-1- to omega-hydroxylated laurate was 3.6, 3.4, and 2.4 for P450 2CAA, P450 2C2, and P450 2C1, respectively. Purified P450 2CAA had a slightly greater apparent molecular weight than expressed P450 2C2 on sodium dodecyl sulfate-polyacrylamide gels. The results clearly establish that rabbit P450 2C1 and 2C2 are arachidonic acid epoxygenases, and they suggest that P450 2CAA and 2C2 are very similar but may not be identical isoforms.


COVID-19: first long-term care facility outbreak in the Netherlands following cross-border introduction from Germany, March 2020.
The Dutch province of Limburg borders the German district of Heinsberg, which had a large cluster of COVID-19 cases linked to local carnival activities before any cases were reported in the Netherlands. However, Heinsberg was not included as an area reporting local or community transmission per the national case definition at the time. In early March, two residents from a long-term care facility (LTCF) in Sittard, a Dutch town located in close vicinity to the district of Heinsberg, tested positive for COVID-19. In this study we aimed to determine whether cross-border introduction of the virus took place by analysing the LTCF outbreak in Sittard, both epidemiologically and microbiologically. Surveys and semi-structured oral interviews were conducted with all present LTCF residents by health care workers during regular points of care for information on new or unusual signs and symptoms of disease. Both throat and nasopharyngeal swabs were taken from residents suspect of COVID-19, based on regional criteria, for the detection of SARS-CoV-2 by Real-time Polymerase Chain Reaction. Additionally, whole genome sequencing was performed using a SARS-CoV-2 specific amplicon-based Nanopore sequencing approach. Moreover, twelve random residents were sampled for possible asymptomatic infections. Out of 99 residents, 46 got tested for COVID-19. Out of the 46 tested residents, nineteen (41%) tested positive for COVID-19, including 3 asymptomatic residents. CT-values for asymptomatic residents seemed higher compared to symptomatic residents. Eleven samples were sequenced, along with three random samples from COVID-19 patients hospitalized in the regional hospital at the time of the LTCF outbreak. All samples were linked to COVID-19 cases from the cross-border region of Heinsberg, Germany. Sequencing combined with epidemiological data was able to virtually prove cross-border transmission at the start of the Dutch COVID-19 epidemic. Our results highlight the need for cross-border collaboration and adjustment of national policy to emerging region-specific needs along borders in order to establish coordinated implementation of infection control measures to limit the spread of COVID-19.


Comparison of changes in extracellular dopamine concentrations in the nucleus accumbens during intravenous self-administration of cocaine or d-amphetamine.
Changes in extracellular concentrations of dopamine (DA) were measured in vivo in the nucleus accumbens of the rat during intravenous self-administration of either cocaine (0.25, 0.5, 1.0mg/infusion) or d-amphetamine (0.05, 0.1, 0.2mg/infusion). Drug intake was limited to 12 self-administered infusions per session for each drug/dose combination. Changes in extra-cellular DA concentrations were measured by two different techniques: chronoamperometry in conjunction with chronically-implanted stearate-modified carbon paste electrodes, or intracerebral microdialysis with off-line analyses using high performance liquid chromatography with electrochemical detection (HPLC-ED). Significant increases in extracellular DA concentrations were observed with both in vivo techniques during self-administration of each dose of cocaine or d-amphetamine. For each drug, the magnitude of change during the first hour of the test session was comparable across doses. However, the change observed over the first 2h period, as measured by microdialysis and HPLC-ED, revealed a dose effect for cocaine, but no dose-response effect for d-amphetamine. The duration of the drug-induced elevation was increased significantly as a function of dose with both cocaine and d-amphetamine. Data from the microdialysis experiments indicated that the high dose of d-amphetamine (0.2mg/infusion) produced a significantly greater increase in extracellular DA concentrations in the nucleus accumbens than did the high dose of cocaine (1.0mg/infusion), but that comparable changes were induced by doses of 0.1mg/infusion of d-amphetamine and 1.0mg/infusion of cocaine, respectively. Each dose of both psychostimulant drugs also produced a significant decrease in dihydroxyphenylacetic acid (DOPAC) levels. The latter finding indicated that the electrochemical signal measured in these studies was not due to the oxidation of DOPAC. These results confirm that self-administration of cocaine or d-amphetamine by the rat is accompanied by a significant increase in extracellular DA concentrations in the nucleus accumbens. The fact that two different psychomotor stimulant drugs of abuse have qualitatively similar neurochemical correlates when self-administered, adds credence to the hypothesis that their reinforcing properties are related to dynamic changes in DA concentrations in the ventral striatal region of the brain.


Sustained conscious access to incidental memories in RSVP.
In visual search of natural scenes, differentiation of briefly fixated but task-irrelevant distractor items from incidental memory is often comparable to explicit memorization. However, many characteristics of incidental memory remain unclear, including the capacity for its conscious retrieval. Here, we examined incidental memory for faces in either upright or inverted orientation using Rapid Serial Visual Presentation (RSVP). Subjects were instructed to detect a target face in a sequence of 8-15 faces cropped from natural scene photographs (Experiment 1). If the target face was identified within a brief time window, the subject proceeded to an incidental memory task. Here, subjects used incidental memory to discriminate between a probe face (a distractor in the RSVP stream) and a novel, foil face. In Experiment 2 we reduced scene-related semantic coherency by intermixing faces from multiple scenes and contrasted incidental memory with explicit memory, a condition where subjects actively memorized each face from the sequence without searching for a target. In both experiments, we measured objective performance (Type 1 AUC) and metacognitive accuracy (Type 2 AUC), revealing sustained and consciously accessible incidental memory for upright and inverted faces. In novel analyses of face categories, we examined whether accuracy or metacognitive judgments are affected by shared semantic features (i.e., similarity in gender, race, age). Similarity enhanced the accuracy of incidental memory discriminations but did not influence metacognition. We conclude that incidental memory is sustained and consciously accessible, is not reliant on scene contexts, and is not enhanced by explicit memorization.


Optimization of adeno-associated viral vector-mediated transduction of the corticospinal tract: comparison of four promoters.
Adeno-associated viral vectors are widely used as vehicles for gene transfer to the nervous system. The promoter and viral vector serotype are two key factors that determine the expression dynamics of the transgene. A previous comparative study has demonstrated that AAV1 displays efficient transduction of layer V corticospinal neurons, but the optimal promoter for transgene expression in corticospinal neurons has not been determined yet. In this paper, we report a side-by-side comparison between four commonly used promoters: the short CMV early enhancer/chicken β actin (sCAG), human cytomegalovirus (hCMV), mouse phosphoglycerate kinase (mPGK) and human synapsin (hSYN) promoter. Reporter constructs with each of these promoters were packaged in AAV1, and were injected in the sensorimotor cortex of rats and mice in order to transduce the corticospinal tract. Transgene expression levels and the cellular transduction profile were examined after 6 weeks. The AAV1 vectors harbouring the hCMV and sCAG promoters resulted in transgene expression in neurons, astrocytes and oligodendrocytes. The mPGK and hSYN promoters directed the strongest transgene expression. The mPGK promoter did drive expression in cortical neurons and oligodendrocytes, while transduction with AAV harbouring the hSYN promoter resulted in neuron-specific expression, including perineuronal net expressing interneurons and layer V corticospinal neurons. This promoter comparison study contributes to improve transgene delivery into the brain and spinal cord. The optimized transduction of the corticospinal tract will be beneficial for spinal cord injury research.


The role of coacervation and phase transitions in the sandcastle worm adhesive system.
Sandcastle worms, Phragmatopoma californica (Fewkes), live along the western coast of North America. Individual worms build tubular shells under seawater by gluing together sandgrains and biomineral particles with a multipart, rapid-set, self-initiating adhesive. The glue comprises distinct sets of condensed, oppositely charged polyelectrolytic components-polyphosphates, polysulfates, and polyamines-that are separately granulated and stored at high concentration in distinct cell types. The pre-organized adhesive modules are secreted separately and intact, but rapidly fuse with minimal mixing and expand into a crack-penetrating complex fluid. Within 30s of secretion into seawater, the fluid adhesive transitions (sets) into a porous solid adhesive joint. The nano- and microporous structure of the foamy solid adhesive contributes to the strength and toughness of the adhesive joint through several mechanisms. A curing agent (catechol oxidase), co-packaged into both types of adhesive granules, covalently cross-links the adhesive and becomes a structural component of the final adhesive joint. The overall effectiveness of the granulated sandcastle glue is more a product of the cellular sorting and packaging mechanisms, the transition from fluid to solid following secretion, and its final biphasic porous structure as it is of its composition or any particular amino acid modification.


Associations of physical activity with childhood asthma, a population study based on the WHO - health behaviour in school-aged children survey.
Asthma in paediatric populations is one of the highest public health concerns. In this study of children and adolescents, we hypothesized that low levels of physical activity (PA) would show associations with asthma that vary by asthma outcome. The objective was to assess whether PA was associated with ever asthma and/or current asthma. Analyses were based on 4824 Danish schoolchildren aged 11-15 years old (48.7% boys) participating in the HBSC survey. The study variables were (1) physician-diagnosed asthma (ever asthma) and (2) physician-diagnosed asthma plus wheezing and/or physician or hospital consultation for wheezing (current asthma). Associations with PA by gender were analysed with multivariate logistic regression using the "variance covariance (vce) cluster" method. The prevalence of ever asthma was 14.3% (boys) and 12.8% (girls), and that of current asthma was 6.8% (boys) and 7.0% (girls). Boys with current asthma showed important differences in low and high PA. We found inverse associations between low PA and ever asthma, odds ratio [95% confidence interval] male: .55 [.30; .99] and female: .47 [.24; .93], and current asthma, male: .27 [.12; .60] (P linear trend = .007) and female: .32 [.11; .94]. The lowest activity levels showed significant inverse associations with asthma, regardless of the definition. For boys, the more stringent (current asthma) of the two paediatric asthma definitions revealed a significant trend with PA, and the direction of associations shifted to positive as weekly PA increased.


Health library for patients in general practice.
We describe the establishment of a health lending library for patients in a suburban general practice. The library is housed in the surgery waiting room and comprises 413 books and 35 audio tapes on topics related to health. Analysis of the first year's use showed 703 borrowings (677 books and 26 audio cassettes). Borrowers ranged the spectrum of social class and educational status but women predominated, and borrowers included many who admitted that they seldom read books. Most borrowings were opportunistic, though some were planned by the patient and a few recommended by a health care professional. Most readers claimed to have learned from their reading and to have found their reading useful. We think that a health library is a useful tool of primary care, particularly for prevention.


Beyond chemotherapy for advanced disease-the role of EGFR and PD-1 inhibitors.
Penile squamous cell carcinoma (SCC) is a rare malignancy with limited treatment options when the tumor is unresectable and/or chemorefractory. Triplet systemic chemotherapy regimens including taxane and cisplatin are recommended, but the response duration can be short and the treatment-related toxicity high. Only a small proportion of patients survive 1 year or longer with the current standard treatment paradigm. Beyond chemotherapy, the use of novel targeted agents, either alone or in combination with traditional chemotherapeutic agents, has appeared to have promising efficacy in patients with platinum-refractory penile cancer. The frequent overexpression of PD-L1 in advanced penile SCC indicates the potential efficacy of PD-1 inhibitors. Upcoming clinical trials using the immune check-point inhibitors may provide exciting landscape and change the paradigm for patients in the future.


The effect of social class on mid-age women's weight control practices and weight gain.
The prevalence of obesity and weight gain is higher in the working class, but we know little about class differences in practices used to control weight. This study examined associations between self-reported measures of social class (upper, middle, or working class), weight control practices, and weight among a cohort of 11,589 mid-aged women (aged 47-52) participating in the Australian Longitudinal Study on Women's Health (ALSWH). A multivariate model tested whether mean weight change over a two-year period differed by social class after adjusting for area of residence, age, education, baseline BMI, and smoking. Working-class women gained significantly more weight at 1.27 (0.07) kg (95% CI: 1.12-1.42) over 2 years, compared with middle/upper-class women at 1.01 (0.07) kg (95% CI: 0.88-1.15). They were significantly more likely to use potentially harmful weight control practices than middle/upper-class women (8.9%) (Chi-squared test=30.65, p<0.0001), and less likely to meet physical activity recommendations. The study provides longitudinal evidence from a nationally representative sample of women that social class is related to weight gain, and to certain weight control practices. The findings have implications for the development of weight gain prevention programs for socially disadvantaged groups.


Antipsoriatic activity of 10-acyl analogues of dithranol (anthralin). I. Phase I clinical trial of 10-propionyl dithranol and 10-butyryl dithranol (butantrone).
Dithranol, 10-propionyl dithranol and 10-butyryl dithranol were compared as regards their antipsoriatic, staining and irritative properties. They were applied under 12-mm Finnchambers on the lesional skin of 17 psoriasis patients every second day for 20 days. In 9 patients their concentration was equimolar (2 mmol/kg of white petrolatum) corresponding to 0.05% dithranol, and in 8 patients the concentrations were 2, 4, and 8 mmol for dithranol, 10-propionyl dithranol and 10-butyryl dithranol, respectively. Without regard to the concentration, they all produced under occlusion almost equal thinning of the lesions, but when a 2 mmol concentration was used, 10-butyryl dithranol (butantrone) showed least staining and irritation on the treated sites.


Os odontoideum: presentation, diagnosis, and treatment in a series of 78 patients.
The most contentious issue in the management of os odontoideum surrounds the decision to attempt atlantoaxial fusion in patients with asymptomatic lesions. The authors examined the clinical presentation and outcome in patients with os odontoideum who underwent surgical stabilization, with an emphasis on 3 patients who initially received conservative treatment and suffered delayed neurological injury. Seventy-eight patients (mean age 20.5 years; median 15 years) were identified in a 17-year retrospective review. The median follow-up period was 14 months (range 1-115 months). Neck pain was the most common symptom (64%), and 56% of patients presented after traumatic injury. Eighteen patients had neurological signs or symptoms at presentation, and an additional 15 had a history of intermittent or prior neurological symptoms. Fifteen patients had undergone > or = 1 attempt at atlantoaxial fusion elsewhere. Seventy-seven patients underwent posterior fusion and rigid screw fixation combined with a graft/wire construct: 75 had C1-2 fusion and 2 had occipitocervical fusion. One patient had an odontoid screw placed. Fusion was achieved in all patients at a median of 4.8 months (range 2-17 months). Approximately 90% of patients had resolution or improvement of their neck pain or neurological symptoms. The authors believe that patients with os odontoideum are at risk for future spinal cord compromise. Forty-four percent of our patients had myelopathic symptoms at referral, and 3 had significant neurological deterioration when a known os odontoideum was left untreated. This risk of late neurological deterioration should be considered when counseling patients. Stabilization using internal screw fixation techniques resulted in 100% fusion, whereas 15% of patients had previously undergone unsuccessful wire and external bracing attempts.


Observational Constraints on the Oxidation of NOx in the Upper Troposphere.
NOx (NOx ≡ NO + NO2) regulates O3 and HOx (HOx ≡ OH + HO2) concentrations in the upper troposphere. In the laboratory, it is difficult to measure rates and branching ratios of the chemical reactions affecting NOx at the low temperatures and pressures characteristic of the upper troposphere, making direct measurements in the atmosphere especially useful. We report quasi-Lagrangian observations of the chemical evolution of an air parcel following a lightning event that results in high NOx concentrations. These quasi-Lagrangian measurements obtained during the Deep Convective Clouds and Chemistry experiment are used to characterize the daytime rates for conversion of NOx to different peroxy nitrates, the sum of alkyl and multifunctional nitrates, and HNO3. We infer the following production rate constants [in (cm(3)/molecule)/s] at 225 K and 230 hPa: 7.2(±5.7) × 10(-12) (CH3O2NO2), 5.1(±3.1) × 10(-13) (HO2NO2), 1.3(±0.8) × 10(-11) (PAN), 7.3(±3.4) × 10(-12) (PPN), and 6.2(±2.9) × 10(-12) (HNO3). The HNO3 and HO2NO2 rates are ∼ 30-50% lower than currently recommended whereas the other rates are consistent with current recommendations to within ±30%. The analysis indicates that HNO3 production from the HO2 and NO reaction (if any) must be accompanied by a slower rate for the reaction of OH with NO2, keeping the total combined rate for the two processes at the rate reported for HNO3 production above.


Cigarette smoke induction of osteopontin (SPP1) mediates T(H)17 inflammation in human and experimental emphysema.
Smoking-related lung diseases are among the leading causes of death worldwide, underscoring the need to understand their pathogenesis and develop new effective therapies. We have shown that CD1a+ antigen-presenting cells (APCs) from lungs of patients with emphysema can induce autoreactive T helper 1 (T(H)1) and T(H)17 cells. Similarly, the canonical cytokines interferon-γ (IFN-γ) and interleukin-17A (IL-17A) are specifically linked to lung destruction in smokers, but how smoke activates APCs to mediate emphysema remains unknown. Here, we show that, in addition to increasing IFN-γ expression, cigarette smoke increased the expression of IL-17A in both CD4+ and γδ T cells from mouse lung. IL-17A deficiency resulted in attenuation of, whereas lack of γδ T cells exacerbated, smoke-induced emphysema in mice. Adoptive transfer of lung APCs isolated from mice with emphysema revealed that this cell population was capable of transferring disease even in the absence of active smoke exposure, a process that was dependent on IL-17A expression. Spp1 (the gene for osteopontin) was highly expressed in the pathogenic lung APCs of smoke-exposed mice and was required for the T(H)17 responses and emphysema in vivo, in part through its inhibition of the expression of the transcription factor Irf7. Thus, the Spp1-Irf7 axis is critical for induction of pathological T(H)17 responses, revealing a major mechanism by which smoke activates lung APCs to induce emphysema and identifying a pathway that could be targeted for therapeutic purposes.


Time-varying cortical activations related to visual-tactile cross-modal links in spatial selective attention.
The neural mechanisms underlying unimodal spatial attention have long been studied, but the cortical processes underlying cross-modal links remain a matter of debate. To reveal the cortical processes underlying the cross-modal links between vision and touch in spatial attention, we recorded magnetoencephalographic (MEG) responses to electrocutaneous stimuli when subjects directed attention to an electrocutaneous or visual stimulus presented randomly in the left or right space. Neural responses recorded around the bilateral sylvian fissures at 85 and 100 ms after the electrocutaneous stimulus were significantly enhanced by spatial attention in both the touch-irrelevant and -relevant modalities. Source analysis revealed that the sylvian responses were generated in the secondary somatosensory cortex (SII). An early response, M50c, generated in the contralateral primary somatosensory cortex (SI), was not modulated by attention. There were no significant attentional changes in the source location or magnetic field distribution, suggesting attentional facilitation of the neural activity in SII itself, rather than a tonic bias effect or overlapping of separate neuronal populations. The results show that spatial attention enhances responses to tactile inputs in SII, independent of sensory modality attended. The underlying mechanism remains to be determined, but may be an increase in gain.


NADPH oxidase deficient mice develop colitis and bacteremia upon infection with normally avirulent, TTSS-1- and TTSS-2-deficient Salmonella Typhimurium.
Infections, microbe sampling and occasional leakage of commensal microbiota and their products across the intestinal epithelial cell layer represent a permanent challenge to the intestinal immune system. The production of reactive oxygen species by NADPH oxidase is thought to be a key element of defense. Patients suffering from chronic granulomatous disease are deficient in one of the subunits of NADPH oxidase. They display a high incidence of Crohn's disease-like intestinal inflammation and are hyper-susceptible to infection with fungi and bacteria, including a 10-fold increased risk of Salmonellosis. It is not completely understood which steps of the infection process are affected by the NADPH oxidase deficiency. We employed a mouse model for Salmonella diarrhea to study how NADPH oxidase deficiency (Cybb (-/-)) affects microbe handling by the large intestinal mucosa. In this animal model, wild type S. Typhimurium causes pronounced enteropathy in wild type mice. In contrast, an avirulent S. Typhimurium mutant (S.Tm(avir); invGsseD), which lacks virulence factors boosting trans-epithelial penetration and growth in the lamina propria, cannot cause enteropathy in wild type mice. We found that Cybb (-/-) mice are efficiently infected by S.Tm(avir) and develop enteropathy by day 4 post infection. Cell depletion experiments and infections in Cybb (-/-) Myd88 (-/-) mice indicated that the S.Tm(avir)-inflicted disease in Cybb (-/-) mice hinges on CD11c(+)CX3CR1(+) monocytic phagocytes mediating colonization of the cecal lamina propria and on Myd88-dependent proinflammatory immune responses. Interestingly, in mixed bone marrow chimeras a partial reconstitution of Cybb-proficiency in the bone marrow derived compartment was sufficient to ameliorate disease severity. Our data indicate that NADPH oxidase expression is of key importance for restricting the growth of S.Tm(avir) in the mucosal lamina propria. This provides important insights into microbe handling by the large intestinal mucosa and the role of NADPH oxidase in maintaining microbe-host mutualism at this exposed body surface.


In vivo carbon-13 nuclear magnetic resonance studies of mammals.
Natural abundance carbon-13 nuclear magnetic resonances (NMR) from human arm and rat tissues have been observed in vivo. These signals arise primarily from triglycerides in fatty tissue. Carbon-13 NMR was also used to follow, in a living rat, the conversion of C-1-labeled glucose, which was introduced into the stomach, to C-1-labeled liver glycogen. The carbon-13 sensitivity and resolution obtained shows that natural abundance carbon-13 NMR will be valuable in the study of disorders in fat metabolism, and that experiments with substrates labeled with carbon-13 can be used to study carbohydrate metabolism in vivo.


Genetic determinants of circulating sphingolipid concentrations in European populations.
Sphingolipids have essential roles as structural components of cell membranes and in cell signalling, and disruption of their metabolism causes several diseases, with diverse neurological, psychiatric, and metabolic consequences. Increasingly, variants within a few of the genes that encode enzymes involved in sphingolipid metabolism are being associated with complex disease phenotypes. Direct experimental evidence supports a role of specific sphingolipid species in several common complex chronic disease processes including atherosclerotic plaque formation, myocardial infarction (MI), cardiomyopathy, pancreatic beta-cell failure, insulin resistance, and type 2 diabetes mellitus. Therefore, sphingolipids represent novel and important intermediate phenotypes for genetic analysis, yet little is known about the major genetic variants that influence their circulating levels in the general population. We performed a genome-wide association study (GWAS) between 318,237 single-nucleotide polymorphisms (SNPs) and levels of circulating sphingomyelin (SM), dihydrosphingomyelin (Dih-SM), ceramide (Cer), and glucosylceramide (GluCer) single lipid species (33 traits); and 43 matched metabolite ratios measured in 4,400 subjects from five diverse European populations. Associated variants (32) in five genomic regions were identified with genome-wide significant corrected p-values ranging down to 9.08x10(-66). The strongest associations were observed in or near 7 genes functionally involved in ceramide biosynthesis and trafficking: SPTLC3, LASS4, SGPP1, ATP10D, and FADS1-3. Variants in 3 loci (ATP10D, FADS3, and SPTLC3) associate with MI in a series of three German MI studies. An additional 70 variants across 23 candidate genes involved in sphingolipid-metabolizing pathways also demonstrate association (p = 10(-4) or less). Circulating concentrations of several key components in sphingolipid metabolism are thus under strong genetic control, and variants in these loci can be tested for a role in the development of common cardiovascular, metabolic, neurological, and psychiatric diseases.


Considerations in the treatment of the node-negative (N0) neck in glottic carcinomas.
Treatment of lymph node negative (N0) glottic carcinoma has raised numerous controversy for decades. Prevention is one of the oldest axioms in medicine. On the other hand, overtreatment can cause unnecessary harm to patients. This retrospective study was performed in 206 patients having glottic cancers with clinically node-negative (N0) necks. The aim of this assessment is to deal with the diagnosis, predictive factors and surgical therapy of occult metastases of squamous cell cancers originating from the glottic region. The examinations were performed in three phases. Preoperative clinical, histological - and in selective cases - imaging were carried out to separate high-risk patients. Intraoperative cases of open surgery after U-shaped skin preparation up to the hyoid bone with direct inspection of jugular lymph node chain (JLNCh) where the neck was staged. The enlarged suspicious nodes were submitted for immediate frozen section. The types of neck dissection were based on the size, shape, number and histological diagnosis of regional nodes. The postoperative additional management was decided according to the results of definitive pathological findings from serial sections of the dissected specimen. Endolaryngeal LASER surgery was carried out in 87 patients based on clinical, histological and imaging criteria. In the course of two years follow-up 2 occult metastases became clinically apparent. At 119 cases open surgery were performed. In 51 patients we could not see enlarged lymph nodes (N< 2 mm) with direct examination, and thus the JLNCh remained intact. In 68 patients elective neck dissections (END) were carried out. In cases of extracapsular spread (ECS) and/or multiple nodal involvements additional radiotherapy was given.


Psychological components of effort sense.
The perception of effort is multidimensional and it is governed by many physiological, psychological, and experiential factors. This paper deals with a discussion of selected psychological states and traits that are known to be correlated with the expression of effort sense. It has been shown that anxiety, somatic perception, depression, and neuroticism are associated with perceived exertion. Extroversion has been found to be inversely correlated with perceived exertion, and positively correlated with preferred exercise intensity. These empirical findings are congruent with theoretical expectations in each case. It has also been found that perception of effort can be increased and decreased in a systematic manner with various psychological interventions such as hypnotic suggestion, dissociative cognitive strategies, and imagery. Changes in effort sense can also be systematically modified by titrating exercise volume (e.g., overtraining, tapering), and this exercise-induced alteration in perception covaries with affective changes. The research reviewed in this paper supports the conclusion that effort sense is best conceptualized as a complex psychobiological construct as originally proposed by Borg three decades ago.


Are lifestyle measures effective in patients with gastroesophageal reflux disease? An evidence-based approach.
Lifestyle modifications are first-line therapy for patients with gastroesophageal reflux disease (GERD). We applied an evidence-based approach to determine the efficacy of lifestyle measures for GERD management. We used PubMed and Ovid to perform a search of the literature published between 1975 and 2004 using the key words heartburn, GERD, smoking, alcohol, obesity, weight loss, caffeine or coffee, citrus, chocolate, spicy food, head of bed elevation, and late-evening meal. Each study was reviewed by 2 reviewers who assigned one of the following ratings: evidence A, randomized clinical trials; evidence B, cohort or case-control studies; evidence C, case reports or flawed clinical trials; evidence D, investigator experience; or evidence E, insufficient information. We screened 2039 studies and identified 100 that were relevant. Only 16 clinical trials examined the impact on GERD (by change in symptoms, esophageal pH variables, or lower esophageal sphincter pressure) of the lifestyle measure. Although there was physiologic evidence that exposure to tobacco, alcohol, chocolate, and high-fat meals decreases lower esophageal sphincter pressure, there was no published evidence of the efficacy of dietary measures. Neither tobacco nor alcohol cessation was associated with improvement in esophageal pH profiles or symptoms (evidence B). Head of bed elevation and left lateral decubitus position improved the overall time that the esophageal pH was less than 4.0 (evidence B). Weight loss improved pH profiles and symptoms (evidence B). Weight loss and head of bed elevation are effective lifestyle interventions for GERD. There is no evidence supporting an improvement in GERD measures after cessation of tobacco, alcohol, or other dietary interventions.


CCAAT/enhancer binding protein is involved in the expression of the tumour necrosis factor gene in human monocytes.
Within the human TNF promoter we have identified two sites at positions -189 and -101 that show C/EBP specific binding of nuclear proteins from cells of the human monocytic line Mono Mac 6. Supershift analysis with anti C/EBP antibodies revealed that the complexes formed consist of both C/EBP alpha and C/EBP beta. When studying reporter constructs with a 5'-deletion series of the TNF promoter in cotransfection experiments with a C/EBP beta expression plasmid, a construct with the -1064 TNF fragment gave 26-fold transactivation, the -630 fragment showed 23-fold transactivation and the -107 fragment (containing the -101 C/EBP binding motif) still gave 16-fold transactivation. Mutagenesis of the -101 site in the -630 construct resulted in a reduction of C/EBP driven transactivation from 26-fold to 7-fold. Finally, when Mono Mac 6 cells were transfected with these constructs, stimulation by LPS induced a 19-fold transactivation in the -630 wild type construct, while the -630 construct carrying the -101 mutation was transactivated only 4-fold. Hence, the data indicate that the -101 C/EBP motif is crucial for TNF gene expression in human monocytes.


Immune responses to lipopolysaccharide challenge in a tropical rodent (Funambulus pennanti): photoperiod entrainment and sex differences.
Annual variation in day length (photoperiod) triggers changes in the immune system of seasonal breeders. The rationale behind this study was to delineate any sex differences in immune responses of photoperiodically entrained animals challenged against lipopolysaccharide (LPS)-induced inflammatory stress. We observed that photoperiodically entrained [short day, SD, 10 h light (L):14 h dark (D); long day, LD, 16 h L:8 h D; and natural day length, NDL, 12 h L:12 h D] male and female Indian palm squirrels, Funambulus pennanti, presented sexual dimorphism in immune status after LPS-induced stress. Females presented high humoral (anti-keyhole limpet hemocyanin immunoglobulin) and cellular immunity (lymphocyte proliferation) compared with the males of all photoperiodic conditions. Female squirrels showed reduced pro-inflammatory cytokine levels (interleukin-1β, interleukin-6, and tumor necrosis factor-α) than the males suggesting their high efficiency to recover from LPS-induced inflammatory stress. Increased duration of melatonin secretion and corticosterone concentration in squirrels experiencing SD evidently supported survival of squirrels as compared with control (NDL) and LD squirrels of both sexes. Decreased immune status in both sexes under LD condition might be due to a short melatonin signal mimicking the LDs of summer. Thus, we infer that photoperiodic entrainment via the levels of melatonin and corticosterone synergistically supported more the survival of female squirrels under LPS-induced stress.


Functional Improvement of Upper and Lower Extremity After Decompression and Neurolysis and Nerve Transfer in a Pediatric Patient with Acute Flaccid Myelitis.
BACKGROUND Acute flaccid myelitis is an emerging polio-like illness mostly affecting young children, characterized by rapid onset of extremity weakness and paralysis in 1 or more limbs. Certain viruses, including enteroviruses such as EV-68, EV-71, poliovirus, and West Nile virus, can cause this disorder. The largest known outbreak of EVD68 in the United States was in the summer of 2014, causing severe respiratory illness and acute flaccid myelitis, mainly in young children. Furthermore, the US Centers for Disease Control and Prevention noted an increase in the number of patients with clinical symptoms of acute flaccid myelitis in 2018, and 134 confirmed cases by December 2018 were reported in the USA. CASE REPORT The patient in our present study was a 5-year-old female who had significant weakness and paralysis in all 4 extremities due to acute flaccid myelitis. EV-D68 had caused this disorder in this patient in August 2014. Conservative management had not helped her condition. Specific areas of concern were both shoulders and biceps, and the femoral and peroneal nerves in both sides. Of these, the right shoulder function was the worst, at less than grade 3. The patient also had marked atrophy and weakness of the right quadricep muscles. The patient underwent surgical treatment and had steady improvements in all 4 extremity functional movements. CONCLUSIONS We demonstrated that decompression, neurolysis, and nerve transfer surgical procedures can be used successfully to correct the paralyzed upper and lower extremity movements in acute flaccid myelitis patients.


Gardnerella vaginalis Subgroups Defined by cpn60 Sequencing and Sialidase Activity in Isolates from Canada, Belgium and Kenya.
Increased abundance of Gardnerella vaginalis and sialidase activity in vaginal fluid is associated with bacterial vaginosis (BV), a common but poorly understood clinical entity associated with poor reproductive health outcomes. Since most women are colonized with G. vaginalis, its status as a normal member of the vaginal microbiota or pathogen causing BV remains controversial, and numerous classification schemes have been described. Since 2005, sequencing of the chaperonin-60 universal target (cpn60 UT) has distinguished four subgroups in isolate collections, clone libraries and deep sequencing datasets. To clarify potential clinical and diagnostic significance of cpn60 subgroups, we undertook phenotypic and molecular characterization of 112 G. vaginalis isolates from three continents. A total of 36 subgroup A, 33 B, 35 C and 8 D isolates were identified through phylogenetic analysis of cpn60 sequences as corresponding to four "clades" identified in a recently published study, based on sequencing 473 genes across 17 isolates. cpn60 subgroups were compared with other previously described molecular methods for classification of Gardnerella subgroups, including amplified ribosomal DNA restriction analysis (ARDRA) and real-time PCR assays designed to quantify subgroups in vaginal samples. Although two ARDRA patterns were observed in isolates, each was observed in three cpn60 subgroups (A/B/D and B/C/D). Real-time PCR assays corroborated cpn60 subgroups overall, but 13 isolates from subgroups A, B and D were negative in all assays. A putative sialidase gene was detected in all subgroup B, C and D isolates, but only in a single subgroup A isolate. In contrast, sialidase activity was observed in all subgroup B isolates, 3 (9%) subgroup C isolates and no subgroup A or D isolates. These observations suggest distinct roles for G. vaginalis subgroups in BV pathogenesis. We conclude that cpn60 UT sequencing is a robust approach for defining G. vaginalis subgroups within the vaginal microbiome.


High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells.
To develop a highly efficacious method for preparation of soluble SARS S-protein using adenovirus vector to meet the requirement for S-protein investigation. The human adenovirus vector was used to express the soluble S-protein (corresponding to 1 approximately 1190 amino acids) fused with Myc/His tag using codon-optimized gene construct in HEK239 cells. The recombinant adenovirus bearing S-protein gene was generated by ligation method. The expressed S-protein with Myc/His tag was purified from culture medium with Ni-NTA agarose beads followed by dialysis. The S-protein was detected by Western blot and its biologic activity was analyzed by binding to Vero cells. Under the conditions of infection dose (MOI of 50) and expression time (48 h), the high-level expression of S-protein was obtained. The expression level was determined to be approximately 75 microg/10(6) cells after purification. Purified soluble S-protein was readily detected by Western blot with anti-Myc antibody and showed the ability to bind to surface of Vero cells,demonstrating that the soluble S-protein could remain the biologic activity in the native molecule. The high-level expression of S-protein in HEK293 cells mediated by adenovirus can be achieved under the optimized expression conditions. The proteins possess the biologic activity, which lays a foundation for further investigation of S-protein biological function.


Investigation into the mechanism of gamma interferon-mediated inhibition of Toxoplasma gondii in murine astrocytes.
Toxoplasma gondii is an obligate intracellular parasite that is a common opportunistic pathogen of the central nervous system in AIDS patients. Gamma interferon (IFN-gamma) alone or in combination with interleukin-1 (IL-1), IL-6, or tumor necrosis factor alpha significantly inhibits the growth of T. gondii in murine astrocytes, suggesting these are important nonimmune effector cells in the brain. Inhibition was found to be independent of a nitric oxide-mediated or tryptophan starvation mechanism. Both reactive oxygen intermediates and iron deprivation are IFN-gamma-mediated mechanisms known to operate against intracellular parasites in other cell types. Astrocytes generated from mice genetically deficient in the production of reactive oxygen intermediates (phox(-/-) mice) were found to inhibit growth of T. gondii when stimulated with IFN-gamma alone or in combination with other cytokines. The reactive oxygen inhibitor catalase and the reactive oxygen scavengers mannitol and thiourea failed to reverse the IFN-gamma-induced inhibition of T. gondii in astrocytes. These data indicate that IFN-gamma-induced inhibition in astrocytes is independent of reactive oxygen intermediates. IFN-gamma-induced inhibition could not be reversed by the addition of iron salts, ferric citrate, ferric nitrate, or ferric transferrin. Pretreatment of astrocytes with desferrioxamine also did not induce the inhibition of T. gondii. These data indicate that the mechanism of IFN-gamma inhibition was not due to iron deprivation. IFN-gamma had no effect on T. gondii invasion of astrocytes, but inhibition of growth and loss of tachyzoite vacuoles were evident in IFN-gamma-treated astrocytes by 24 h after invasion. Overall, these data suggest that IFN-gamma-activated astrocytes inhibit T. gondii by an as-yet-unknown mechanism.


Antibodies to RNA polymerase III in systemic sclerosis detected by ELISA.
To determine serological and clinical variables associated with anti-RNA polymerase III (RNAP-III) antibodies in patients with systemic sclerosis (SSc) using a new ELISA method. Sera from 242 patients with SSc were collected from 14 Canadian clinics. Control sera were from 287 blood donors, and 42 patients with infectious disease, 30 with rheumatoid arthritis (RA), and 30 with systemic lupus erythematosus (SLE). Antibodies to RNAP-III were detected by an ELISA kit and antibodies to other cellular antigens were identified by indirect immunofluorescence (IIF) on HEp-2 cell substrate, line immunoassay, immunoprecipitation of recombinant protein, and addressable laser bead immunoassay (ALBIA). Anti-RNAP-III antibodies were detected in 47/242 (19.4%) SSc sera, 0% RA and SLE sera, 1/287 blood donor sera, and 2/42 infectious disease sera. Diffuse disease (59.5%) was more common than limited disease (36.1%) in the anti-RNAP-III-positive patients (p = 0.006) and there was an association between the presence of anti-RNAP-III and kidney and joint/tendon involvement, but there was no association with a nucleolar IIF pattern, lung involvement, or other clinical indicators. There was a negative association between the presence of anti-RNAP-III antibodies and anticentromere by IIF (p = 0.00004) and anti-Scl-70 by ALBIA (p = 0.0005) and line immunoassay (p = 0.003), suggesting a virtually exclusive presence of these antibodies in SSc. Anti-RNAP-III autoantibodies were found in nearly 20% of SSc patients but in less than 1% of controls, thus detection of this antibody is a useful marker to help diagnose SSc. As well, this antibody has prognostic utility, since it is associated with scleroderma renal crisis and the diffuse cutaneous form of SSc.


Affordable in-house antiretroviral drug resistance assay with good performance in non-subtype B HIV-1.
The introduction of antiretroviral (ARV) therapy in resource-poor settings is effective in suppressing HIV-1 replication and prolonging life of infected individuals. This has led to a demand for affordable HIV-1 drug resistance assays, since treatment failure due to development of drug resistance is common. This study developed and evaluated an affordable "in-house" genotyping assay to monitor HIV-1 drug resistance in Africa, particularly South Africa. An "in-house" assay using automated RNA extraction, and subtype C specific PCR and sequencing primers was developed and successfully evaluated 396 patient samples (viral load ranges 1000-1.6 million RNA copies/ml). The "in-house" assay was validated by comparing sequence data and drug resistance profiles from 90 patient and 10 external quality control samples to data from the ViroSeq HIV-1 Genotyping kit. The "in-house" assay was more efficient, amplifying all 100 samples, compared to 91 samples using Viroseq. The "in house" sequences were 99.2% homologous to the ViroSeq sequences, and identical drug resistance mutation profiles were observed in 96 samples. Furthermore, the "in-house" assay genotyped 260 of 295 samples from seven African sites, where 47% were non-subtype C. Overall, the newly validated "in-house" drug resistance assay is suited for use in Africa as it overcomes the obstacle of subtype diversity.


The etiology of potential small-bowel bleeding depending on patient's age and gender.
Whether the etiology of potential small-bowel bleeding depends on the age and gender of the patient is not yet fully understood. A total of 1953 patients who underwent video capsule endoscopy (VCE) to evaluate potential small-bowel bleeding and were registered in the Capsule Endoscopy Nationwide Database Registry from 2003 to 2014 were eligible for this study. VCE findings and the etiology of small-bowel bleeding were analyzed by age and gender. The diagnostic yield of VCE was 48.4% (95% CI: 46.2%-50.6%) and the diagnosis rate of etiology of potential small-bowel bleeding was 61.4% (95% CI: 59.2%-63.6%). The etiology of potential small-bowel bleeding depends on the age and gender of the patient. Crohn's disease and small-bowel diverticular diseases were more prevalent etiology of potential small-bowel bleeding in the young adults group (< 40 years) whereas angiodysplasia was revealed to be a most common etiology in elderly group (≥ 60 years), reaching statistical significance (p<0.00152) by Bonferroni correction. The etiology of potential small-bowel bleeding depends on the age of the patient. Thus, an individualized lesion-specific diagnostic approach based on age might be needed for patients with potential small-bowel bleeding.


Modulation of human mesenchymal and pluripotent stem cell behavior using biophysical and biochemical cues: A review.
In vitro manipulation of human stem cells is a critical process in regenerative medicine and cellular therapies. Strategies and methods to maintain stem cells and direct them into specific lineages are ongoing challenges in these fields. To date, a number of studies have reported that besides biochemical stimulation, biophysical cues in the form of surface patterning and external stimulation also influence stem cell attachment, proliferation, and differentiation, and can be used in cell reprogramming and the maintenance of pluripotency. While biochemical cues are generally effective and easy to deliver, biophysical cues have many other advantages for scalability as they are cost efficient, have a longer lifetime, and can be easily defined. However, different protocols and cell sources utilized in a variety of studies have led to difficulties in obtaining clear conclusions about the effects of the biophysical environment on stem cells. In addition, the examination of different types of external stimulation is time consuming and limited by available fabrication techniques, resulting in a delay in commercialization and clinical applications. In this review, we aim to summarize the most important biophysical cues and methods for the culture of human stem cells, including mesenchymal and pluripotent stem cells, to facilitate their adoption in stem cell biology. The standard classical protocols of using biochemical cues will also be discussed for comparison. We believe that combining biochemical and biophysical stimulation has the greatest potential to generate functionally mature cells at a scalable and inexpensive rate for diverse applications in regenerative medicine and cell therapy. Biotechnol. Bioeng. 2017;114: 260-280. © 2016 Wiley Periodicals, Inc.


The chromatin structure of the GAL1 promoter forms independently of Reb1p in Saccharomyces cerevisiae.
Positive and negative regulation of the GAL1 promoter of the yeast Saccharomyces cerevisiae results from a network of interactions between transcription factors and chromatin. In this study we used footprinting procedures to characterize these interactions in vivo. DNase I analysis of the GAL1 upstream activating sequence (UAS(GAL1/10)) showed expected Gal4 activator protein binding during growth in galactose, and also revealed binding of the Reb1 protein (Reb1p) during growth in glucose. In addition, we mapped to nucleotide resolution a positioned nucleosome that, in the inactive promoter, packages DNA between the UAS(GAL1/10) and the GAL1 TATA sequence, leaving both of these elements nucleosome free. The nucleosome footprint was lost when the promoter was activated. Surprisingly, mutation of the Reb1p binding site had no effect on nucleosome positioning or on the kinetics or extent of activation or repression of either the GAL1 or GAL10 promoters under any of the conditions assayed.


Physiologic effects of steep positioning in the surgical intensive care unit.
Ten hemodynamically stable patients requiring mechanical ventilation for radiographically symmetric acute lung disease were studied during steep lateral positioning and continuous rotation in a Roto Rest kinetic treatment bed. There were no significant hemodynamic or ventilatory differences among the four positions (supine, right side down, left side down, and rotating). In four patients, arterial oxygen pressure (PaO2) decreased 16% to 49% in the lateral position when compared with the supine position. Continuous rotation restored the PaO2 toward the supine value in each patient. In six patients, PaO2 increased 11% to 35% during lateral positioning. In five of the six patients, the increase in PaO2 differed between sides, suggesting asymmetric lung disease. Continuous rotation did not significantly alter the PaO2 from the supine values in these patients. Adverse effects on oxygenation caused by positional changes may be reversed by continuous rotation using the Roto Rest kinetic bed.


Fine structure recombinational analysis of cloned genes using yeast transformation.
We describe a general method for analyzing the genetic fine structure of plasmid-borne genes in yeast. Previously we had reported that a linearized plasmid is efficiently rescued by recombination with a homologous restriction fragment when these are co-introduced by DNA-mediated transformation of yeast. Here, we show that a mutation can be localized to a small DNA interval when members of a deletion series of wild-type restriction fragments are used in the rescue of a linearized mutant plasmid. The resolution of this method is to at least 30 base pairs and is limited by the loss of a wild-type marker with proximity to a free DNA end. As a means for establishing the nonidentity of two mutations, we determined the resolution of two-point crosses with a mutant linearized plasmid and a mutant homologous restriction fragment. Recombination between mutations separated by as little as 100 base pairs was detected. Moreover, the results indicate that exchange within a marked interval results primarily from one of two single crossovers that repair the linearized plasmid. These approaches to mapping the genetic fine structure of plasmids should join existing methods in a robust approach to the mutational analysis of gene structure in yeast.


Equal performance of aspiration and stent retriever thrombectomy in daily stroke treatment.
Mechanical thrombectomy with stent retrievers has proved to be safe and effective in endovascular treatment of acute ischemic stroke. Direct aspiration has shown revascularization rates comparable to those of stent retrievers in the recent ASTER and COMPASS trials. However, the efficacy of aspiration in routine clinical practice has not yet been shown. To show that aspiration has clinical and technical outcomes equal to those of stent retriever thrombectomy in daily clinical practice. We analysed data of patients with a large vessel occlusion of the anterior circulation registered in the Dutch MR CLEAN Registry between March 2014 and June 2016. Primary outcome was functional outcome measured with the modified Rankin Scale (mRS) score. Secondary outcomes were reperfusion grade, periprocedural complication rate, and procedure duration. Association of treatment technique with functional outcome was estimated with univariable and multivariable ordinal logistic regression analysis and expressed as a common OR (cOR) for a shift towards better outcome on the mRS. As first-line treatment, 207 of 1175 patients (17.6%) were treated with direct aspiration, and 968 (82.4%) by a stent retriever. We observed no differences in functional outcome (adjusted cOR=1.020 (95% CI 0.68 to 1.52)) and periprocedural complications. Successful reperfusion (extended Thrombolysis in Cerebral Infarction ≥2b) was similar. Duration of the procedure was shorter with aspiration (57 min (IQR 35-73) vs 70 min (IQR 47-95), p<0.0001). Direct aspiration shows clinical outcomes equal to those of stent retriever thrombectomy in our large multicenter real-life cohort. We found no difference in complication rates and shorter procedure times for aspiration.


Effects of haemodialysis and continuous ambulatory peritoneal dialysis on abnormalities of ion transport in vivo in patients with chronic renal failure.
1. We have studied Na+/K+ pump activity in vivo in three groups of subjects: patients with chronic renal failure not receiving maintenance dialysis, patients being treated by either haemodialysis or continuous ambulatory peritoneal dialysis, and matched control subjects. 2. To do this we have measured the changes in plasma and intraerythrocytic rubidium concentrations after an oral load of rubidium chloride, having previously shown that changes in the disposition of rubidium measured in this way reflect changes in the activity of the Na+/K+ pump in vivo. 3. Erythrocyte rubidium uptake was significantly reduced both in ten patients with chronic renal failure not receiving maintenance dialysis and in 12 patients being treated by haemodialysis, when compared with 31 healthy control subjects. In contrast, erythrocyte rubidium uptake was not altered in 13 patients treated by continuous ambulatory peritoneal dialysis. There was also a significantly reduced rate constant for erythrocyte rubidium uptake in patients with undialysed chronic renal failure (0.66 h-1) and in those treated by haemodialysis (0.78 h-1), whereas in patients treated by continuous ambulatory peritoneal dialysis the rate constant for erythrocyte rubidium uptake was not significantly different from control values (1.36 h-1 and 1.41 h-1, respectively). 4. These findings are consistent with a reversal of the inhibition of erythrocyte Na+/K+ pump activity in vivo found in chronic renal failure by continuous ambulatory peritoneal dialysis, but not by haemodialysis. This difference may be due to the failure of haemodialysis to clear a circulating inhibitor of Na+, K(+)-ATPase or to the rapid re-accumulation of such an inhibitor after haemodialysis.(ABSTRACT TRUNCATED AT 250 WORDS)


Optimizing millisecond time scale near-infrared emission in polynuclear chrome(III)-lanthanide(III) complexes.
This work illustrates a simple approach for optimizing long-lived near-infrared lanthanide-centered luminescence using trivalent chromium chromophores as sensitizers. Reactions of the segmental ligand L2 with stoichiometric amounts of M(CF(3)SO(3))(2) (M = Cr, Zn) and Ln(CF(3)SO(3))(3) (Ln = Nd, Er, Yb) under aerobic conditions quantitatively yield the D(3)-symmetrical trinuclear [MLnM(L2)(3)](CF(3)SO(3))(n) complexes (M = Zn, n = 7; M = Cr, n = 9), in which the central lanthanide activator is sandwiched between the two transition metal cations. Visible or NIR irradiation of the peripheral Cr(III) chromophores in [CrLnCr(L2)(3)](9+) induces rate-limiting intramolecular intermetallic Cr→Ln energy transfer processes (Ln = Nd, Er, Yb), which eventually produces lanthanide-centered near-infrared (NIR) or IR emission with apparent lifetimes within the millisecond range. As compared to the parent dinuclear complexes [CrLn(L1)(3)](6+), the connection of a second strong-field [CrN(6)] sensitizer in [CrLnCr(L2)(3)](9+) significantly enhances the emission intensity without perturbing the kinetic regime. This work opens novel exciting photophysical perspectives via the buildup of non-negligible population densities for the long-lived doubly excited state [Cr*LnCr*(L2)(3)](9+) under reasonable pumping powers.


Effect of Preoperative Opioid Exposure on Healthcare Utilization and Expenditures Following Elective Abdominal Surgery.
To examine the extent to which preoperative opioid use is correlated with healthcare utilization and costs following elective surgical procedures. Morbidity and mortality associated with prescription opioid use is escalating in the United States. The extent to which chronic opioid use influences postoperative outcomes following elective surgery is not well understood. Truven Health Marketscan Databases were used to identify adult patients who underwent elective abdominal surgery between June 2009 and December 2012 (n = 200,005). Generalized linear regression was used to determine the effect of preoperative opioid use on postoperative healthcare utilization (length of stay, 30-d readmission, and discharge destination) and cost (hospital stay, 90-, 180-, and 365-d) after adjusting for number of comorbidities, psychological conditions, and demographic characteristics. In this cohort, 8.8% of patients used opioids preoperatively. Compared with non-users, patients using opioids preoperatively were more likely to have a longer hospital stay (2.9 d vs. 2.5 d, P <0.001) and were more likely to be discharged to a rehabilitation facility (3.6% vs. 2.5%, P <0.001), adjusting for covariates. Preoperative opioid use was also correlated with a greater rate of 30-day readmission (4.5% vs. 3.6%, P <0.001) and overall greater expenditures at 90- ($12036.60 vs. $3863.40, P <0.001), 180- ($16973.70 vs. $6790.60, P <0.001), and 365- ($25495.70 vs. $12113.80, P <0.001) days following surgery, adjusted for covariates. Additionally, dose-effects were observed regarding readmission, discharge destination, and late healthcare expenditures. Preoperative interventions focused on opioid cessation and alternative analgesics may improve the safety and efficiency of elective surgery among chronic opioid users.


Language comprehension in boys with fragile X syndrome and boys with Down syndrome.
Fragile X syndrome (FXS) is the most common known inherited cause of intellectual disability, yet very few studies have explored the language comprehension skills of children with FXS. We examined the receptive vocabulary, grammatical morphology and syntax skills of boys with FXS (who were additionally classified as having autism, autism spectrum, or no autism) and compared them to boys with Down syndrome (DS) and typically developing (TD) boys at similar non-verbal developmental levels. The Vocabulary, Grammatical Morphology, and Elaborated Phrases and Sentences subtests of the Test for Auditory Comprehension of Language - 3rd Edition (TACL-3) were administered annually up to three times to assess the language comprehension skills of 35 boys with FXS without autism, 24 boys with FXS with autism spectrum, 19 boys with FXS with autism, 45 boys with DS and 40 TD boys at similar non-verbal cognitive levels. After controlling for non-verbal cognition and maternal education levels, we found that the three groups of boys with FXS did not differ from each other but scored lower than the TD boys in language comprehension. The boys with DS scored lower in language comprehension than boys with FXS without autism and TD boys. For all of the groups, scores for receptive vocabulary, grammatical morphology and syntax did not differ. Boys with FXS and boys with DS differed in receptive language levels, demonstrating unique language profiles for each syndrome. Language comprehension appears to be an important area to target in assessment and intervention for both populations.


Cost-effectiveness of anticoagulants for suspected heparin-induced thrombocytopenia in the United States.
Despite the availability of multiple nonheparin anticoagulants for the treatment of heparin-induced thrombocytopenia (HIT), few data are available comparing the cost-effectiveness of these agents. This analysis is particularly important when considering differences in the risk of adverse effects, routes of administration, requirements for phlebotomy and laboratory monitoring, and overall drug costs. We conducted a cost-effectiveness analysis of argatroban, bivalirudin, and fondaparinux for the treatment of suspected HIT from the institutional perspective. A 3-arm decision-tree model was developed that employs standard practices for anticoagulation monitoring. We incorporated published data on drug efficacy and probability of HIT-related thromboembolism and major bleeding. We considered both institutional costs and average wholesale price (AWP) and performed probabilistic sensitivity analyses (PSA) to address any uncertainty in model parameters. Using institutional costs, fondaparinux prevailed over both argatroban and bivalirudin in terms of cost ($151 vs $1250 and $1466, respectively) and adverse events averted (0.9989 vs 0.9957 and 0.9947, respectively). Results were consistent when AWP was used, with fondaparinux being less expensive ($555 vs $3081 and $2187, respectively) and more effective in terms of adverse events averted (0.9989 vs 0.9957 and 0.9947, respectively). The PSA confirmed our findings using both institutional costs and AWP. In conclusion, fondaparinux subcutaneous injection afforded significant advantages in terms of cost savings and adverse events averted compared with IV argatroban or bivalirudin infusions. Our data strongly suggest potential cost savings with fondaparinux and underscore the critical need for larger clinical studies of fondaparinux in the treatment of suspected HIT.


Orthogonal switching of self-sorting processes in a stimuli-responsive library of cucurbit[8]uril complexes.
The self-sorting processes in dynamic libraries of cucurbit[8]uril complexes can be switched in an orthogonal way by external stimuli. Protonated phenylpyridine guests form a 2 : 1 homoternary π-donor-π-acceptor complex, while deprotonation makes it a partner for ethylviologen in a 1 : 1 : 1 heteroternary complex. Reduction of the viologen instead generates a 2 : 1 homoternary complex of viologen radical-cations.


Analysis of the resolution limitations of peptide identification algorithms.
Proteome identification using peptide-centric proteomics techniques is a routinely used analysis technique. One of the most powerful and popular methods for the identification of peptides from MS/MS spectra is protein database matching using search engines. Significance thresholding through false discovery rate (FDR) estimation by target/decoy searches is used to ensure the retention of predominantly confident assignments of MS/MS spectra to peptides. However, shortcomings have become apparent when such decoy searches are used to estimate the FDR. To study these shortcomings, we here introduce a novel kind of decoy database that contains isobaric mutated versions of the peptides that were identified in the original search. Because of the supervised way in which the entrapment sequences are generated, we call this a directed decoy database. Since the peptides found in our directed decoy database are thus specifically designed to look quite similar to the forward identifications, the limitations of the existing search algorithms in making correct calls in such strongly confusing situations can be analyzed. Interestingly, for the vast majority of confidently identified peptide identifications, a directed decoy peptide-to-spectrum match can be found that has a better or equal match score than the forward match score, highlighting an important issue in the interpretation of peptide identifications in present-day high-throughput proteomics.


Targeting Serotonin1A Receptors for Treating Chronic Pain and Depression.
The association of chronic pain with depression is becoming increasingly recognized. Treating both the conditions together is essential for an effective treatment outcome. In this regard, it is important to identify a shared mechanism involved in the association of chronic pain with depression. Central serotonin (5-hydroxytryptamine; 5-HT) neurotransmission has long been known to participate in the processing of signals related to pain. It also plays a key role in the pathogenesis and treatment of depression. Although functional responses to serotonin are mediated via the activation of multiple receptor types and subtypes, the 5-HT1A subtype is involved in the processing of nociception as well as the pathogenesis and treatment of depression. This receptor is located presynaptically, as an autoreceptor, on the perikaryon and dendritic spines of serotonin-containing neurons. It is also expressed as a heteroreceptor on neurons receiving input from serotonergic neurons. This article targets the 5-HT1A receptors to show that indiscriminate activation of pre and postsynaptic 5-HT1A receptors is likely to produce no therapeutic benefits; biased activation of the 5-HT heteroreceptors may be a useful strategy for treating chronic pain and depression individually as well as in a comorbid condition.


Staphylococcus epidermidis polysaccharide intercellular adhesin induces IL-8 expression in human astrocytes via a mechanism involving TLR2.
Staphylococcus epidermidis is an opportunistic biofilm-forming pathogen associated with neurosurgical device-related meningitis. Expression of the polysaccharide intercellular adhesin (PIA) on its surface promotes S. epidermidis biofilm formation. Here we investigated the pro-inflammatory properties of PIA against primary and transformed human astrocytes. PIA induced IL-8 expression in a dose- and/or time-dependent manner from U373 MG cells and primary normal human astrocytes. This effect was inhibited by depletion of N-acetyl-beta-d-glucosamine polymer from the PIA preparation with Lycopersicon esculentum lectin or sodium meta-periodate. Expression of dominant-negative versions of the TLR2 and TLR4 adaptor proteins MyD88 and Mal in U373 MG cells inhibited PIA-induced IL-8 production. Blocking IL-1 had no effect. PIA failed to induce IL-8 production from HEK293 cells stably expressing TLR4. However, in U373 MG cells which express TLR2, neutralization of TLR2 impaired PIA-induced IL-8 production. In addition to IL-8, PIA also induced expression of other cytokines from U373 MG cells including IL-6 and MCP-1. These data implicate PIA as an important immunogenic component of the S. epidermidis biofilm that can regulate pro-inflammatory cytokine production from human astrocytes, in part, via TLR2.


Effect of wavelength on in vivo images of the human cone mosaic.
In images of the human fundus, the fraction of the total returning light that comes from the choroidal layers behind the retina increases with wavelength [Appl. Opt. 28, 1061 (1989); Vision Res. 36, 2229 (1996)]. There is also evidence that light originating behind the receptors is not coupled into the receptor waveguides en route to the pupil [S. A. Burns et al., Noninvasive Assessment of the Visual System, Vol. 11 of 1997 Trends in Optics and Photonics Series, D. Yager, ed. (Optical Society of America, 1997), p. al; Invest. Ophthalmol. Visual Sci. 38, 1657 (1997)]. These observations imply that the contrast of images of the cone mosaic should be greatly reduced with increasing wavelength. This hypothesis was tested by imaging the light distributions in both the planes of the photoreceptors and the pupil at three wavelengths, 550, 650, and 750 nm, with the Rochester adaptive optics ophthalmoscope. Surprisingly, the contrast of the retinal images varied only slightly with wavelength. Furthermore, the ratio of the receptorally guided component to the total reflected light measured in the pupil plane was found to be similar at each wavelength, suggesting that, throughout this wavelength range, the scattered light from the deeper layers in the retina is guided through the receptors on its return path to the pupil.


Epidemiology of urethral strictures.
Urethral stricture disease is relatively common and is associated with a significant financial cost and potentially debilitating outcomes. Understanding urethral stricture epidemiology is important to identify risk factors associated with the etiology or progression of the disease. This understanding may lead to better treatments and preventative measures that could ameliorate disease severity, produce better health outcomes, and reduce expenditures. We performed a comprehensive review of urethral stricture disease based on available published case series, identified gaps in knowledge of this disease, and recommend future directions for research.


The effectiveness of sealants in managing caries lesions.
A barrier to providing sealants is concern about inadvertently sealing over caries. This meta-analysis examined the effectiveness of sealants in preventing caries progression. We searched electronic databases for comparative studies examining caries progression in sealed permanent teeth. We used a random-effects model to estimate percentage reduction in the probability of caries progression in sealed vs. unsealed carious teeth. Six studies, including 4 randomized-controlled trials (RCT) judged to be of fair quality, were included in the analysis (384 persons, 840 teeth, and 1090 surfaces). The median annual percentage of non-cavitated lesions progressing was 2.6% for sealed and 12.6% for unsealed carious teeth. The summary prevented fraction for RCT was 71.3% (95%CI: 52.8%-82.5, no heterogeneity) up to 5 years after placement. Despite variation among studies in design and conduct, sensitivity analysis found the effect to be consistent in size and direction. Sealing non-cavitated caries in permanent teeth is effective in reducing caries progression.


Cost-effectiveness of endovascular thrombectomy with alteplase versus endovascular thrombectomy alone for acute ischemic stroke secondary to large vessel occlusion.
Recent randomized trials have suggested that endovascular thrombectomy (EVT) alone may provide similar functional outcomes as the current standard of care, EVT combined with intravenous alteplase treatment, for acute ischemic stroke secondary to large vessel occlusion. We conducted an economic evaluation of these 2 therapeutic options. We constructed a decision analytic model with a hypothetical cohort of 1000 patients to assess the cost-effectiveness of EVT with intravenous alteplase treatment versus EVT alone for acute ischemic stroke secondary to large vessel occlusion from both the societal and public health care payer perspectives. We used studies and data published in 2009-2021 for model inputs, and acquired cost data for Canada and China, representing high- and middle-income countries, respectively. We calculated incremental cost-effectiveness ratios (ICERs) using a lifetime horizon and accounted for uncertainty using 1-way and probabilistic sensitivity analyses. All costs are reported in 2021 Canadian dollars. In Canada, the difference in quality-adjusted life-years (QALYs) gained between EVT with alteplase and EVT alone was 0.10 from both the societal and health care payer perspectives. The difference in cost was $2847 from a societal perspective and $2767 from the payer perspective. In China, the difference in QALYs gained was 0.07 from both perspectives, and the difference in cost was $1550 from the societal perspective and $1607 from the payer perspective. One-way sensitivity analyses showed that the distributions of modified Rankin Scale scores at 90 days after stroke were the most influential factor on ICERs. For Canada, compared to EVT alone, the probability that EVT with alteplase would be cost-effective at a willingness-to-pay threshold of $50 000 per QALY gained was 58.7% from a societal perspective and 58.4% from a payer perspective. The corresponding values for at a willingness-to-pay threshold of $47 185 (3 times the Chinese gross domestic product per capita in 2021) were 65.2% and 67.4%. For patients with acute ischemic stroke due to large vessel occlusion eligible for immediate treatment with both EVT alone and EVT with intravenous alteplase treatment, it is uncertain whether EVT with alteplase is cost-effective compared to EVT alone in Canada and China.


Ex Situ Perfusion of Hearts Donated After Euthanasia: A Promising Contribution to Heart Transplantation.
Organ donation after euthanasia is performed in an increasing number of countries. In this donation after circulatory death procedure, it has not been possible to donate the heart. Recent literature, however, reports positive results of heart donation after circulatory death. Therefore, patients who donate organs following euthanasia might be suitable candidates for heart donation. We want to confirm this assumption by sharing the results of 2 cases of heart donation following euthanasia with ex situ subnormothermic heart preservation. Our aim is to raise awareness of the potential of heart donation following euthanasia for both clinical transplantation and research. The data of 2 consecutive heart donations following euthanasia were collected prospectively. Informed consent was obtained from the patients themselves for heart donation for research purposes. An acellular oxygenated subnormothermic machine perfusion strategy was used to preserve both donor hearts. Subsequently, the hearts were evaluated on a normothermic perfusion machine using a balloon in the left ventricle. Heart donation following euthanasia was feasible without significant changes in existing retrieval protocols. Duration of machine perfusion preservation was 408 and 432 minutes, for heart 1 and 2, respectively. For heart 1, developed pressure (Pdev) was 119 mm Hg, maximal rate of pressure rise (dP/dtmax), and fall (dP/dtmin) were 1524 mm Hg/s and -1057 mm Hg/s, respectively. For heart 2, Pdev was 142 mm Hg, dP/dtmax was 1098 mm Hg/s, and dP/dtmin was -802 mm Hg/s. Hearts donated following euthanasia are highly valuable for research purposes and can have sufficient quality to be transplanted. With the implementation of ex situ heart perfusion, patients who are to donate their organs following euthanasia should also be able to donate their hearts. The complex combination of euthanasia and heart donation is ethically sound and surgically feasible and can contribute to shortening the heart transplant waiting list.


CALBC silver standard corpus.
The CALBC initiative aims to provide a large-scale biomedical text corpus that contains semantic annotations for named entities of different kinds. The generation of this corpus requires that the annotations from different automatic annotation systems be harmonized. In the first phase, the annotation systems from five participants (EMBL-EBI, EMC Rotterdam, NLM, JULIE Lab Jena, and Linguamatics) were gathered. All annotations were delivered in a common annotation format that included concept identifiers in the boundary assignments and that enabled comparison and alignment of the results. During the harmonization phase, the results produced from those different systems were integrated in a single harmonized corpus ("silver standard" corpus) by applying a voting scheme. We give an overview of the processed data and the principles of harmonization--formal boundary reconciliation and semantic matching of named entities. Finally, all submissions of the participants were evaluated against that silver standard corpus. We found that species and disease annotations are better standardized amongst the partners than the annotations of genes and proteins. The raw corpus is now available for additional named entity annotations. Parts of it will be made available later on for a public challenge. We expect that we can improve corpus building activities both in terms of the numbers of named entity classes being covered, as well as the size of the corpus in terms of annotated documents.


The effect of magnesium supplementation on blood pressure in individuals with insulin resistance, prediabetes, or noncommunicable chronic diseases: a meta-analysis of randomized controlled trials.
Background: To our knowledge, the effect of magnesium supplementation on blood pressure (BP) in individuals with preclinical or noncommunicable diseases has not been previously investigated in a meta-analysis, and the findings from randomized controlled trials (RCTs) have been inconsistent.Objective: We sought to determine the pooled effect of magnesium supplementation on BP in participants with preclinical or noncommunicable diseases.Design: We identified RCTs that were published in English before May 2017 that examined the effect of magnesium supplementation on BP in individuals with preclinical or noncommunicable diseases through PubMed, ScienceDirect, Cochrane, clinicaltrials.gov, SpringerLink, and Google Scholar databases as well as the reference lists from identified relevant articles. Random- and fixed-effects models were used to estimate the pooled standardized mean differences (SMDs) with 95% CIs in changes in BP from baseline to the end of the trial in both systolic blood pressure (SBP) and diastolic blood pressure (DBP) between the magnesium-supplementation group and the control group.Results: Eleven RCTs that included 543 participants with follow-up periods that ranged from 1 to 6 mo (mean: 3.6 mo) were included in this meta-analysis. The dose of elemental magnesium that was used in the trials ranged from 365 to 450 mg/d. All studies reported BP at baseline and the end of the trial. The weighted overall effects indicated that the magnesium-supplementation group had a significantly greater reduction in both SBP (SMD: -0.20; 95% CI: -0.37, -0.03) and DBP (SMD: -0.27; 95% CI: -0.52, -0.03) than did the control group. Magnesium supplementation resulted in a mean reduction of 4.18 mm Hg in SBP and 2.27 mm Hg in DBP.Conclusion: The pooled results suggest that magnesium supplementation significantly lowers BP in individuals with insulin resistance, prediabetes, or other noncommunicable chronic diseases.


Rapid bacterial genome sequencing: methods and applications in clinical microbiology.
The recent advances in sequencing technologies have given all microbiology laboratories access to whole genome sequencing. Providing that tools for the automated analysis of sequence data and databases for associated meta-data are developed, whole genome sequencing will become a routine tool for large clinical microbiology laboratories. Indeed, the continuing reduction in sequencing costs and the shortening of the 'time to result' makes it an attractive strategy in both research and diagnostics. Here, we review how high-throughput sequencing is revolutionizing clinical microbiology and the promise that it still holds. We discuss major applications, which include: (i) identification of target DNA sequences and antigens to rapidly develop diagnostic tools; (ii) precise strain identification for epidemiological typing and pathogen monitoring during outbreaks; and (iii) investigation of strain properties, such as the presence of antibiotic resistance or virulence factors. In addition, recent developments in comparative metagenomics and single-cell sequencing offer the prospect of a better understanding of complex microbial communities at the global and individual levels, providing a new perspective for understanding host-pathogen interactions. Being a high-resolution tool, high-throughput sequencing will increasingly influence diagnostics, epidemiology, risk management, and patient care.


Characterization of genome-wide association-identified variants for atrial fibrillation in African Americans.
Despite a greater burden of risk factors, atrial fibrillation (AF) is less common among African Americans than European-descent populations. Genome-wide association studies (GWAS) for AF in European-descent populations have identified three predominant genomic regions associated with increased risk (1q21, 4q25, and 16q22). The contribution of these loci to AF risk in African American is unknown. We studied 73 African Americans with AF from the Vanderbilt-Meharry AF registry and 71 African American controls, with no history of AF including after cardiac surgery. Tests of association were performed for 148 SNPs across the three regions associated with AF, and 22 SNPs were significantly associated with AF (P<0.05). The SNPs with the strongest associations in African Americans were both different from the index SNPs identified in European-descent populations and independent from the index European-descent population SNPs (r(2)<0.40 in HapMap CEU): 1q21 rs4845396 (odds ratio [OR] 0.30, 95% confidence interval [CI] 0.13-0.67, P = 0.003), 4q25 rs4631108 (OR 3.43, 95% CI 1.59-7.42, P = 0.002), and 16q22 rs16971547 (OR 8.1, 95% CI 1.46-45.4, P = 0.016). Estimates of European ancestry were similar among cases (23.6%) and controls (23.8%). Accordingly, the probability of having two copies of the European derived chromosomes at each region did not differ between cases and controls. Variable European admixture at known AF loci does not explain decreased AF susceptibility in African Americans. These data support the role of 1q21, 4q25, and 16q22 variants in AF risk for African Americans, although the index SNPs differ from those identified in European-descent populations.


Pharmacological Modulation of Serotonin Levels in Zebrafish Larvae: Lessons for Identifying Environmental Neurotoxicants Targeting the Serotonergic System.
This study examines the effects of acute pharmacological modulation of the serotonergic system over zebrafish larvae's cognitive, basic, and defense locomotor behaviors, using a medium to high throughput screening assay. Furthermore, the relationship between behavior, enzyme activity related to neurotransmitter metabolism, neurotransmitter levels, and gene expression was also determined. Modulation of larvae serotonergic system was accomplished by 24 h exposure to single and opposite pharmacodynamics co-exposure to three model psychopharmaceuticals with antagonistic and agonistic serotonin signaling properties: 2.5 mM 4-Chloro-DL-phenylalanine (PCPA) and 5 µM deprenyl and 0.5 µM fluoxetine, respectively. Similar behavioral outcome was observed for deprenyl and fluoxetine, which was reflected as hypolocomotion, decrease in larvae defensive responses, and cognitive impairment. Contrarily, PCPA induced hyperlocomotion and increase in larvae escape response. Deprenyl exposure effects were more pronounced at a lower level of organization than fluoxetine, with complete inhibition of monoamine oxidase (MAO) activity, dramatic increase of 5-HT and dopamine (DA) levels, and downregulation of serotonin synthesis and transporter genes. PCPA showed mainly effects over serotonin and dopamine's main degradation metabolites. Finally, co-exposure between agonistic and antagonist serotonin signaling drugs reviled full recovery of zebrafish impaired locomotor and defense responses, 5-HT synthesis gene expression, and partial recovery of 5-HT levels. The findings of this study suggest that zebrafish larvae can be highly sensitive and a useful vertebrate model for short-term exposure to serotonin signaling changes.


Joint effects of nutrient addition and enemy exclusion on exotic plant success.
Worldwide, ecosystems are increasingly dominated by exotic plant species, a shift hypothesized to result from numerous ecological factors. Two of these, increased resource availability and enemy release, may act in concert to increase exotic success in plant communities (Resource-Enemy Release Hypothesis, R-ERH). To test this, we manipulated the availability of soil nutrients and access of vertebrate herbivores, insect herbivores, and fungal pathogens to intact grassland communities containing both native and exotic species. Our results supported both conditions necessary for R-ERH. First, exotics were less damaged than natives, experiencing less foliar damage (insect herbivory and fungal disease) than native species, particularly in communities where soil nutrients were added. Second, fertilization increased foliar damage on native species, but not exotic species. As well as fulfilling both conditions for R-ERH, these results demonstrate the importance of considering the effects of resource availability when testing for enemy release. When both conditions are fulfilled, R-ERH predicts that increasing resource availability will increase exotic abundance only in the presence of enemies. Our results fully supported this prediction for vertebrate herbivores: fertilization increased exotic cover only in communities exposed to vertebrate herbivores. Additionally, the prediction was partially supported for insect herbivores and fungal pathogens, excluding these enemies reduced exotic cover as predicted, but inconsistent with R-ERH, this effect occurred only in unfertilized communities. These results highlight the need to consider the influence of multiple enemy guilds on community processes like exotic plant invasions. Moreover, this study experimentally demonstrates that resource availability and natural enemies can jointly influence exotic success in plant communities.


Socio-cultural influences on the behaviour of South Asian women with diabetes in pregnancy: qualitative study using a multi-level theoretical approach.
Diabetes in pregnancy is common in South Asians, especially those from low-income backgrounds, and leads to short-term morbidity and longer-term metabolic programming in mother and offspring. We sought to understand the multiple influences on behaviour (hence risks to metabolic health) of South Asian mothers and their unborn child, theorise how these influences interact and build over time, and inform the design of culturally congruent, multi-level interventions. Our sample for this qualitative study was 45 women of Bangladeshi, Indian, Sri Lankan, or Pakistani origin aged 21-45 years with a history of diabetes in pregnancy, recruited from diabetes and antenatal services in two deprived London boroughs. Overall, 17 women shared their experiences of diabetes, pregnancy, and health services in group discussions and 28 women gave individual narrative interviews, facilitated by multilingual researchers, audiotaped, translated, and transcribed. Data were analysed using the constant comparative method, drawing on sociological and narrative theories. Key storylines (over-arching narratives) recurred across all ethnic groups studied. Short-term storylines depicted the experience of diabetic pregnancy as stressful, difficult to control, and associated with negative symptoms, especially tiredness. Taking exercise and restricting diet often worsened these symptoms and conflicted with advice from relatives and peers. Many women believed that exercise in pregnancy would damage the fetus and drain the mother's strength, and that eating would be strength-giving for mother and fetus. These short-term storylines were nested within medium-term storylines about family life, especially the cultural, practical, and material constraints of the traditional South Asian wife and mother role and past experiences of illness and healthcare, and within longer-term storylines about genetic, cultural, and material heritage - including migration, acculturation, and family memories of food insecurity. While peer advice was familiar, meaningful, and morally resonant, health education advice from clinicians was usually unfamiliar and devoid of cultural meaning. 'Behaviour change' interventions aimed at preventing and managing diabetes in South Asian women before and during pregnancy are likely to be ineffective if delivered in a socio-cultural vacuum. Individual education should be supplemented with community-level interventions to address the socio-material constraints and cultural frames within which behavioural 'choices' are made.


An amperometric needle-type glucose sensor tested in rats and man.
An amperometric glucose-measuring 25 gauge (0.5 mm diameter) needle-type sensor has been developed using a glucose oxidase and dimethyl ferrocene paste behind a semi-permeable membrane situated over a window in the needle. Electron transfer results in direct current generation. Sensors have been tested subcutaneously in the abdomen both in anaesthetized rats (40 sensors, 11 rats) and in normal, conscious man (20 sensors, 10 subjects). In rats the blood glucose was modulated by glucose and by insulin infusion. In man the glucose concentrations were rapidly changed by use of a glucose clamp at 12 mmol/l plasma concentration for 2 h, after which the glucose returned to normal. In rats the median correlation between glucose change was 0.83 with an interquartile range from 0.70 to 0.92, and in man the median correlation was 0.80 with an interquartile range 0.67 to 0.86. Hysteresis, a measure of the accuracy on the upswing and downswing, was not a problem and cross-correlation showed no phase-lag. There were quantitative differences between in vitro calibration and the performance in vivo, reflecting the different conditions of use. The current in response to a glucose concentration was stable over 6.0 h in rats and 4.5 h in man.