Reducing hospital readmissions through primary care practice transformation.
To assess the impact of a multicomponent intervention on 30-day hospital readmissions in a group of primary care practices that undertook practice transformation, compared with rates in usual-care practices that admitted patients to the same hospital service. Four primary care clinics enhanced patient care coordination with care managers and inpatient care teams, and developed and used hospital readmission reports to monitor readmission rates. Patient readmissions to the hospital were analyzed over a 12-month period from May 2012 through April 2013, among patients who fell into 2 groups. Group 1 patients were those cared for by the primary care clinics that implemented transformation activities and who were admitted to the hospital associated with the practices. Group 2 patients were seen at clinics in the same catchment area that did not undertake any known practice redesign activities (usual care group). A total of 961 patients were included in analyses; 685 (71.3%) were in Group 1, and 276 (28.7%) were in Group 2. Readmissions among Group 1 patients decreased from 27% to 7.1% (P=.02), and readmissions in Group 2 were variable with a nonsignificant trend (P=.53). The unadjusted regression model that compared the interaction between Group 1 and Group 2 patients found a significant difference in readmissions (P=.05). Developing a multicomponent intervention appears to have a significant impact on reducing hospital readmissions. Primary care groups seeking to reduce hospital readmissions should consider implementing similar processes.


Human cerebellum: surface-assisted cortical parcellation and volumetry with magnetic resonance imaging.
We describe a system of surface-assisted parcellation (SAP) of the human cerebellar cortex derived from neural systems functional and behavioral anatomy. This system is based on MRI and preserves the unique morphologic and topographic features of the individual cerebellum. All major fissures of the cerebellum were identified and traced in the flattened representation of the cerebellar cortex using the program "FreeSurfer." Parcellation of the cerebellar cortex followed using the fissure information in conjunction with landmarks using the program "Cardviews" to create 64 gyral-based cerebellar parcellation units. Computer-assisted algorithms enable the execution of the cerebellar parcellation procedure as well as volumetric measurements and topographic localization. The SAP technique makes it possible to represent multimodal structural and functional imaging data on the flattened surface of the cerebellar cortex as illustrated in one functional MRI experiment.


Cities and peripheries.
This article addresses a methodological problem of urban history faced with the current environmental crisis that urges us to think of humans as 'geological' agents. It suggests that the concept of the uncanny that pushes our understanding of spatio-temporality may be a useful device for approaching the methodological need to reconcile what we can and cannot experience/visualize. Viewing the mapping projects around Calcutta in the late eighteenth and early nineteenth centuries through the lens of the uncanny offers us the possibility of such a reconciliation. It enables us to see the landscape as a product of multiple spatio-temporal modes, and loosens the grip of the current urban vocabulary on our imagination of cities.


Public Health Policy in Support of Insurance Coverage for Smoking Cessation Treatments.
Insurance coverage for evidence-based smoking cessation treatments (SCTs) promotes uptake and reduces smoking rates. Published studies in this area are based in the US where employers are the primary source of health insurance. In Ontario, Canada, publicly funded healthcare does not cover SCTs, but it can be supplemented with employer-sponsored benefit plans. This study explores factors affecting the inclusion/exclusion of smoking cessation (SC) benefits. In total, 17 interviews were conducted with eight employers (auto, retail, banking, municipal and university industries), four health insurers, two government representatives and three advisors/consultants. Overall, SCT coverage varied among industries; it was inconsistently restrictive and SCT differed by coverage amount and length of use. Barriers impeding coverage included the lack of the following: Canadian-specific return on investment (ROI), SC cost information, employer demand, government regulations/incentives and employee awareness of and demand. A Canadian evidence-based calculation of ROI for SC coupled with government incentives and public education may be needed to promote uptake of SCT coverage by employers.


Nonpharmacological Management of Gout and Hyperuricemia: Hints for Better Lifestyle.
We reviewed lifestyle factors that influence serum uric acid levels and risk of gout flare, and how to improve their deleterious effects. Since obesity increases uric acid and weight gain increases gout risk, weight reduction by daily exercise and limiting intake of excess calories is recommended. However, strenuous exercise, which causes adenine nucleotide degradation; starvation, which decreases uric acid excretion; and dehydration may raise the level of uric acid in serum and trigger gout. Increased intake of purine-rich foods, such as meat and seafood, raise the level of uric acid in serum and is associated with increased risk of gout, whereas dairy products, especially low-fat types, are associated with a lower risk of gout. Also, heavy alcohol drinking raises the uric acid level and increases the risk of gout through adenine nucleotide degradation and lactate production. Sweet fruits and soft drinks containing fructose should be moderated, since fructose may raise uric acid and increase gout risk through uric acid production and/or decreased excretion. On the other hand, the Mediterranean diet is recommended for gout patients, since it may also help prevent hyperuricemia. Furthermore, coffee and vitamin C supplementation could be considered as preventive measures, as those can lower serum uric acid levels as well as the risk of gout.


Feasibility of pedigree recording and genetic selection in village sheep flocks of smallholder farmers.
Pedigree recording and genetic selection in village flocks of smallholder farmers have been deemed infeasible by researchers and development workers. This is mainly due to the difficulty of sire identification under uncontrolled village breeding practices. A cooperative village sheep-breeding scheme was designed to achieve controlled breeding and implemented for Menz sheep of Ethiopia in 2009. In this paper, we evaluated the reliability of pedigree recording in village flocks by comparing genetic parameters estimated from data sets collected in the cooperative village and in a nucleus flock maintained under controlled breeding. Effectiveness of selection in the cooperative village was evaluated based on trends in breeding values over generations. Heritability estimates for 6-month weight recorded in the village and the nucleus flock were very similar. There was an increasing trend over generations in average estimated breeding values for 6-month weight in the village flocks. These results have a number of implications: the pedigree recorded in the village flocks was reliable; genetic parameters, which have so far been estimated based on nucleus data sets, can be estimated based on village recording; and appreciable genetic improvement could be achieved in village sheep selection programs under low-input smallholder farming systems.


Sustained self-regulation of energy intake. Loss of weight in overweight subjects. Maintenance of weight in normal-weight subjects.
Dietary restraint is largely unsuccessful for controlling obesity. As an alternative, subjects can easily be trained to reliably recognize sensations of initial hunger (IH) a set of physiological sensations which emerge spontaneously, not necessarily at planned mealtimes, and may be the afferent arm of a homeostatic system of food intake regulation. Previously we have reported that IH is associated with blood glucose concentration (BG) below 81.8 mg/dL (4.55 mmol/l), (low blood glucose, LBG), and that a pattern of meals in which IH is present pre-meal (IHMP) improved insulin sensitivity, HbA1c and other cardiovascular risk factors. Here we report the effect upon weight in overweight and normal weight subjects. To investigate whether the IHMP is associated with sustained loss of weight in overweight subjects over a 5 month period. SEVENTY FOUR OVERWEIGHT SUBJECTS (OW: BMI > 25) and 107 normal weight (NW) subjects were randomly allocated to either trained (OW: N = 51; NW N = 79) or control (OW: N = 23; NW: N = 28) groups. All subjects were allocated post-randomization into either low or high mean pre-meal BG groups (LBG and HBG groups) using a demarcation point of 81.8 mg/dL. A SIGNIFICANT LONGITUDINAL DECREASE WAS FOUND IN BODY WEIGHT (TRAINED NW: -2.5 +/- 4.6 kg; OW -6.7 +/- 4.5 kg; controls: NW +3.5 +/- 4.0 kg and OW -3.4 +/- 4.0 kg; P = 0.006 and 0.029) and in energy intake, mean BG, standard deviation of diary BG (BG as recorded by subjects' 7-day diary), BMI, and arm and leg skin-fold thickness in (OW and NW) HBG subjects. OW LBG subjects significantly decreased body weight (trained: -4.0 +/- 2.4 kg; controls: -0.4 +/- 3.7 kg; P = 0.037). 26 NW LBG subjects showed no longitudinal difference after training as did 9 control subjects. Over a 5 month period the IHMP resulted in significant loss of weight in OW subjects compared to controls practicing dietary restraint. NW subjects maintained weight overall, however NW HBG subjects also lost weight compared to controls.


The expression of sarcomeric muscle-specific contractile protein genes in BC3H1 cells: BC3H1 cells resemble skeletal myoblasts that are defective for commitment to terminal differentiation.
The BC3H1 cell line has been used widely as a model for studying regulation of muscle-related proteins, such as the acetylcholine receptor, myokinase, creatine kinase, and actin. These cells, derived from a nitrosourea-induced mouse brain neoplasm, have some of the morphological characteristics of smooth muscle and have been shown to express the vascular smooth muscle isoform of alpha-actin. To provide further information about the contractile protein phenotype of BC3H1 and to gain additional insights into the possible tissue of origin of these cells, we have examined the expression of a battery of contractile protein genes. During rapid growth, subconfluent BC3H1 cells express the nonmuscle isoform of alpha-tropomyosin (alpha-Tm) and the nonsarcomeric isoforms of myosin heavy and light chains (MHCs and MLCs, respectively), but do not express troponin T(TnT). However, when BC3H1 cells differentiate in response to incubation in serum-deprived medium or upon approaching confluence, they express TnT as well as sarcomeric muscle isoforms of MHC, MLC 2 and 3, alpha-Tm, and alpha-actin. These results suggest that BC3H1 is a skeletal muscle cell line of ectodermal origin that is defective for commitment to terminal differentiation.


Urea recycling in sheep: effects of intake.
The effect of intake on urea production, entry into the digestive tract and return of N to the ornithine cycle was studied in four sheep. Each sheep received 0.6, 1.2 and 1.8 x estimated maintenance energy intake quantities of grass pellets for 9 d. After 4 d of adjustment, N balance measurements were conducted between days 5 and 8. From day 7 to day 9 animals were continuously infused, via the jugular vein, with [15N15N]urea and three urine samples were collected at approximately 2 h intervals 48-54 h after the start of infusion. Total urea and enrichments of [15N15N]- and [14N15N]urea in the urine samples were determined. Urea production was calculated from the isotopic dilution of [15N15N]urea and entry into the gastrointestinal tract (GIT) obtained from the difference between this and urinary urea elimination. Urea which enters the GIT undergoes hydrolysis to liberate NH3 which may be reabsorbed and enter the ornithine cycle in which case the product is [14N15N]urea, based on the probabilities of labelled and unlabelled N providing ureagenic precursors. The quantity of urea-N which returns to the ornithine cycle from the GIT can thus be calculated. Existing models based on this approach yield overestimates of the fate of individual urea molecules due to a failure to allow for multiple recycling of [14N15N]urea species through the GIT. Refinements introduced to cover this resulted in a 33-48% reduction in calculated return of label for the current study. The present model also predicted that 95% of the label movements across the GIT could be accommodated by three or fewer entries and returns of urea-N and 99% by recycling for a maximum of six occasions. Urea-N production increased with intake (P < 0.001) and exceeded digestible N values at all intakes. Urea which entered the digestive tract, both in absolute terms (P < 0.001) and as a proportion of production (0.62, 0.69, 0.73; P = 0.027), increased with intake. The proportion of entry into the digestive tract which was returned to the ornithine cycle remained reasonably constant (0.37-0.41) across all intakes but the absolute amount increased (5.6, 9.2 and 15.0 g N/d; P < 0.001) with intake. If allowance is included for losses of 15N in faeces then the approach offers a relatively simple means of estimating anabolic reuse of urea by digestive tract micro-organisms and can complement data obtained from the technically more demanding arterio-venous and multiple-isotope techniques used hitherto.


Common tea formulations modulate in vitro digestive recovery of green tea catechins.
Epidemiological evidence suggests a role for tea catechins in reduction of chronic disease risk. However, stability of catechins under digestive conditions is poorly understood. The objective of this study was to characterize the effect of common food additives on digestive recovery of tea catechins. Green tea water extracts were formulated in beverages providing 4.5, 18, 23, and 3.5 mg per 100 mL epicatechin (EC), epigallocatechin (EGC), epigallocatechin-gallate (EGCG), and epicatechin-gallate (ECG), respectively. Common commercial beverage additives; citric acid (CA), BHT, EDTA, ascorbic acid (AA), milk (bovine, soy, and rice), and citrus juice (orange, grapefruit, lemon, and lime) were formulated into finished tea beverages at incremental dosages. Samples were then subjected to in vitro digestion simulating gastric and small intestinal conditions with pre- and post-digestion catechin profiles assessed by HPLC. Catechin stability in green tea was poor with <20% total catechins remaining post-digestion. EGC and EGCG were most sensitive with less, not double equals 10% recovery. Teas formulated with 50% bovine, soy, and rice milk increased total catechin recovery significantly to 52, 55, and 69% respectively. Including 30 mg AA in 250 mL of tea beverage significantly (p<0.05) increased catechin recovery of EGC, EGCG, EC, and ECG to 74, 54, 82, and 45% respectively. Juice preparation resulted in the highest recovery of any formulation for EGC (81-98%), EGCG (56-76%), EC (86-95%), and ECG (30-55%). These data provide evidence that tea consumption practices and formulation factors likely impact catechin digestive recovery and may result in diverse physiological profiles.


Acquisition of Antibodies Against Endothelial Protein C Receptor-Binding Domains of Plasmodium falciparum Erythrocyte Membrane Protein 1 in Children with Severe Malaria.
Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) mediates parasite sequestration in postcapillary venules in P. falciparum malaria. PfEMP1 types can be classified based on their cysteine-rich interdomain region (CIDR) domains. Antibodies to different PfEMP1 types develop gradually after repeated infections as children age, and antibodies to specific CIDR types may confer protection. Levels of immunoglobulin G to 35 recombinant CIDR domains were measured by means of Luminex assay in acute-stage (baseline) and convalescent-stage plasma samples from Papua New Guinean children with severe or uncomplicated malaria and in healthy age-matched community controls. At baseline, antibody levels were similar across the 3 groups. After infection, children with severe malaria had higher antibody levels than those with uncomplicated malaria against the endothelial protein C receptor (EPCR) binding CIDRα1 domains, and this difference was largely confined to older children. Antibodies to EPCR-binding domains increased from presentation to follow-up in severe malaria, but not in uncomplicated malaria. The acquisition of antibodies against EPCR-binding CIDRα1 domains of PfEMP1 after a severe malaria episode suggest that EPCR-binding PfEMP1 may have a role in the pathogenesis of severe malaria in Papua New Guinea.


Evaluation of the second generation of a bioresorbable everolimus drug-eluting vascular scaffold for treatment of de novo coronary artery stenosis: six-month clinical and imaging outcomes.
The first generation of the bioresorbable everolimus drug-eluting vascular scaffold showed signs of shrinkage at 6 months, which largely contributed to late luminal loss. Nevertheless, late luminal loss was less than that observed with bare metal stents. To maintain the mechanical integrity of the device up to 6 months, the scaffold design and manufacturing process of its polymer were modified. Quantitative coronary angiography, intravascular ultrasound with analysis of radiofrequency backscattering, and as an optional assessment, optical coherence tomography (OCT) were performed at baseline and at a 6-month follow-up. Forty-five patients successfully received a single bioresorbable everolimus drug-eluting vascular scaffold. One patient had postprocedural release of myocardial enzyme without Q-wave occurrence; 1 patient with OCT-diagnosed disruption of the scaffold caused by excessive postdilatation was treated 1 month later with a metallic drug-eluting stent. At follow-up, 3 patients declined recatheterization, 42 patients had quantitative coronary angiography, 37 had quantitative intravascular ultrasound, and 25 had OCT. Quantitative coronary angiography disclosed 1 edge restenosis (1 of 42; in-segment binary restenosis, 2.4%). At variance with the ultrasonic changes seen with the first generation of bioresorbable everolimus drug-eluting vascular scaffold at 6 months, the backscattering of the polymeric struts did not decrease over time, the scaffold area was reduced by only 2.0% with intravascular ultrasound, and no change was noted with OCT. On an intention-to-treat basis, the late lumen loss amounted to 0.19±0.18 mm with a limited relative decrease in minimal luminal area of 5.4% on intravascular ultrasound. OCT showed at follow-up that 96.8% of the struts were covered and that malapposition of at least 1 strut, initially observed in 12 scaffolds, was detected at follow-up in only 3 scaffolds. Mean neointimal growth measured by OCT between and on top of the polymeric struts equaled 1.25 mm(2), or 16.6% of the scaffold area. Modified manufacturing process of the polymer and geometric changes in the polymeric platform have substantially improved the medium-term performance of this new generation of drug-eluting scaffold to become comparable to those of current drug eluting stents. URL: http://clinicaltrials.gov. Unique identifier: NCT00856856.


Sensing interstitial glucose to nudge active lifestyles (SIGNAL): feasibility of combining novel self-monitoring technologies for persuasive behaviour change.
Increasing physical activity (PA) reduces the risk of developing diabetes, highlighting the role of preventive medicine approaches. Changing lifestyle behaviours is difficult and is often predicated on the assumption that individuals are willing to change their lifestyles today to reduce the risk of developing disease years or even decades later. The self-monitoring technologies tested in this study will present PA feedback in real time, parallel with acute physiological data. Presenting the immediate health benefits of being more physically active may help enact change by observing the immediate consequences of that behaviour. The present study aims to assess user engagement with the self-monitoring technologies in individuals at moderate-to-high risk of developing type 2 diabetes. 45 individuals with a moderate-to-high risk, aged ≥40 years old and using a compatible smartphone, will be invited to take part in a 7-week protocol. Following 1 week of baseline measurements, participants will be randomised into one of three groups: group 1- glucose feedback followed by biobehavioural feedback (glucose plus PA); group 2-PA feedback followed by biobehavioural feedback; group 3-biobehavioural feedback. A PA monitor and a flash glucose monitor will be deployed during the intervention. Participants will wear both devices throughout the intervention but blinded to feedback depending on group allocation. The primary outcome is the level of participant engagement and will be assessed by device use and smartphone usage. Feasibility will be assessed by the practicality of the technology and screening for diabetes risk. Semistructured interviews will be conducted to explore participant experiences using the technologies. ISRCTN17545949. Registered on 15/05/2017.


Epidemiologic evaluation of Nhanes for environmental Factors and periodontal disease.
Periodontitis is a chronic inflammation that destroys periodontal tissues caused by the accumulation of bacterial biofilms that can be affected by environmental factors. This report describes an association study to evaluate the relationship of environmental factors to the expression of periodontitis using the National Health and Nutrition Examination Study (NHANES) from 1999-2004. A wide range of environmental variables (156) were assessed in patients categorized for periodontitis (n = 8884). Multiple statistical approaches were used to explore this dataset and identify environmental variable patterns that enhanced or lowered the prevalence of periodontitis. Our findings indicate an array of environmental variables were different in periodontitis in smokers, former smokers, or non-smokers, with a subset of specific environmental variables identified in each population subset. Discriminating environmental factors included blood levels of lead, phthalates, selected nutrients, and PCBs. Importantly, these factors were found to be coupled with more classical risk factors (i.e. age, gender, race/ethnicity) to create a model that indicated an increased disease prevalence of 2-4 fold across the sample population. Targeted environmental factors are statistically associated with the prevalence of periodontitis. Existing evidence suggests that these may contribute to altered gene expression and biologic processes that enhance inflammatory tissue destruction.


Chromatic dispersion compensation in coherent transmission system using digital filters.
We present a comparative analysis of three popular digital filters for chromatic dispersion compensation: a time-domain least mean square adaptive filter, a time-domain fiber dispersion finite impulse response filter, and a frequency-domain blind look-up filter. The filters are applied to equalize the chromatic dispersion in a 112-Gbit/s non-return-to-zero polarization division multiplexed quadrature phase shift keying transmission system. The characteristics of these filters are compared by evaluating their applicability for different fiber lengths, their usability for dispersion perturbations, and their computational complexity. In addition, the phase noise tolerance of these filters is also analyzed.


Melatonin and Ischemic Stroke: Mechanistic Roles and Action.
Stroke is one of the most devastating neurological disabilities and brain's vulnerability towards it proves to be fatal and socio-economic loss of millions of people worldwide. Ischemic stroke remains at the center stage of it, because of its prevalence amongst the several other types attacking the brain. The various cascades of events that have been associated with stroke involve oxidative stress, excitotoxicity, mitochondrial dysfunction, upregulation of Ca(2+) level, and so forth. Melatonin is a neurohormone secreted by pineal and extra pineal tissues responsible for various physiological processes like sleep and mood behaviour. Melatonin has been implicated in various neurological diseases because of its antioxidative, antiapoptotic, and anti-inflammatory properties. We have previously reviewed the neuroprotective effect of melatonin in various models of brain injury like traumatic brain injury and spinal cord injury. In this review, we have put together the various causes and consequence of stroke and protective role of melatonin in ischemic stroke.


Specialized olfactory receptor neurons mediating intra- and interspecific chemical communication in leafminer moths Eriocrania spp. (Lepidoptera: Eriocraniidae).
We performed a physiological and morphological characterization of sensilla auricillica in male Eriocrania semipurpurella moths. Each auricillic sensillum contained three olfactory receptor neurons. Responding neurons (87 of 139) could be grouped into five physiological types. Type 1 responded to (R,Z)-6-nonen-2-ol and type 2 to its enantiomer (S,Z)-6-nonen-2-ol, both of which are pheromone components of E. semipurpurella. Type 3 responded to both (R)-heptan-2-ol and (R,Z)-4-hepten-2-ol, which are pheromone components of the sympatric species E. cicatricella. Types 4 and 5 responded to the ketones (Z)-6-nonen-2-one and/or nonan-2-one, which are found in the pheromone glands of female E. semipurpurella. Field-trapping showed that type 3 receptor neurons mediate strongly antagonistic effects of (R)-heptan-2-ol and (R,Z)-4-hepten-2-ol on E. semipurpurella, while nonan-2-one should possibly be included as a synergist in the sex pheromone blend of this species. The attraction of E. cicatricella and E. sparrmannella to compounds mixed with the pheromone blend of E. semipurpurella shows that the pheromone components of E. semipurpurella have little or no antagonistic effects on these species. The morphology and physiology of eriocraniid pheromone sensilla are very similar to those found in the order Trichoptera (caddisflies), suggesting a homology between pheromone detection systems in the two sister orders Lepidoptera and Trichoptera.


Computerized tomography-based quality assurance tool for proton range compensators.
The authors have developed an automatic quality assurance method based on computerized tomography (CT) data for physically manufactured proton range compensators. Eight range compensators used for proton therapy at their institution were scanned using CT and their depth distributions were compared with data provided by a proton treatment planning system (TPS). The depth difference (DD), distance to agreement (DTA), and a composite analysis (CA) indicating regions that passed either of the tests were used for depth verification. When the tolerance limits were set at 3 mm for DD and 1 mm for DTA, the average percentages of points exceeding the acceptance criteria for DD, DTA, and CA for the eight compensators were 9.0%, 5.3% and 3.2%, respectively. In general, the percentages of points exceeding the acceptance criteria were much higher in DD than in DTA analysis and mismatch of depth was greater in regions with high depth gradients. This is considered to arise mainly because of systematic errors such as the physical size of the drill, slight bending or slipping of the drill bit near high gradient regions, the limit of CT resolution, and the uncertainty in CT number. When the tolerance limit of DTA was increased from 1 to 2 mm, the average percentage of points exceeding the acceptance criteria for DTA was reduced from 5.3% to 0.9%. Their results show that depth comparison of proton range compensators based on CT images can provide more systematic data than does the current method, in which a representative group of drill points is measured manually.


Polymer escape from a confining potential.
The rate of escape of polymers from a two-dimensionally confining potential well has been evaluated using self-avoiding as well as ideal chain representations of varying length, up to 80 beads. Long timescale Langevin trajectories were calculated using the path integral hyperdynamics method to evaluate the escape rate. A minimum is found in the rate for self-avoiding polymers of intermediate length while the escape rate decreases monotonically with polymer length for ideal polymers. The increase in the rate for long, self-avoiding polymers is ascribed to crowding in the potential well which reduces the free energy escape barrier. An effective potential curve obtained using the centroid as an independent variable was evaluated by thermodynamic averaging and Kramers rate theory then applied to estimate the escape rate. While the qualitative features are well reproduced by this approach, it significantly overestimates the rate, especially for the longer polymers. The reason for this is illustrated by constructing a two-dimensional effective energy surface using the radius of gyration as well as the centroid as controlled variables. This shows that the description of a transition state dividing surface using only the centroid fails to confine the system to the region corresponding to the free energy barrier and this problem becomes more pronounced the longer the polymer is. A proper definition of a transition state for polymer escape needs to take into account the shape as well as the location of the polymer.


St Theresa's dart and a case of religious ecstatic epilepsy.
: We report a case of ecstatic seizures with religious overtones in a patient whose semiology resembles that of Saint Theresa of Avila (1515 to 1582), and review the anatomical basis of this phenomenon. : Seizures with an emotional component are typically associated with negative sensations. Ecstatic seizures, particularly those with a religious character, are rarely reported, although they hold an important place in history, as a number of religious figures have been posited to be epileptic. Here, we present a patient with ecstatic seizures whose semiology and religious overtones resemble those of Saint Theresa of Avila (1515 to 1582), and discuss the anatomical basis of this phenomenon. : Case report. : Evaluation of the patient demonstrated right temporal encephalomalacia on magnetic resonance imaging of the brain. Electroencephalogram showed focal slowing and poorly defined sharp waves in the right frontotemporal region. This area of pathology concurs with prior studies that localize ecstatic seizures to the anterior insula or temporal lobes. : Ecstatic seizures may involve the right temporal region. Although interpretation of ecstatic seizures is subject to individual variation, a similar localization and our patient's religious background may explain the resemblance between the semiology of his seizures, and that of Saint Theresa's visions as described almost 5 centuries ago.


Movement patterns and genetic diversity of wild and reintroduced common dormice, Muscardinus avellanarius.
Movement is an important life history trait that can have an impact on local adaptation, and other evolutionary phenomena. We used a combination of nestbox survey data and genetic techniques (genotyping at 10 microsatellite loci) to quantify patterns of movement in common dormice Muscardinus avellanarius at two distinct sites in the UK: 1) Bontuchel (a natural population) and 2) Wych (captive-bred individuals that were reintroduced to this site), over three consecutive years (2006-2008). Both methods revealed a consistent pattern of sex-biased movement (movements by adult males and females) in both populations that allowed isolation-by-distance genetic structure to develop within 1 km. The similarity of data from captive-bred and natural individuals indicated that ex situ programing has not significantly altered the natural movement behavior of common dormice; consequently, the two populations could be managed with the same conservation strategies. We also found that the reintroduced dormice in Wych maintained relatively high levels of genetic diversity. This first report of movement patterns in reintroduced and natural populations of M. avellanarius combining genetic and field-survey data highlights the role of genetic studies in the investigation of ecological behaviour and for conservation management.


Molecular characterization of the staphylococcal multidrug resistance export protein QacC.
The QacC polypeptide is a member of a family of small membrane proteins which confer resistance to toxic compounds. The staphylococcal qacC gene confers resistance to toxic organic cations via proton-dependent export. The membrane topology of the QacC polypeptide was investigated by constructing and analyzing a series of qacC-phoA and qacC-lacZ fusions. From these analyses, most of the predicted features of the QacC protein were verified, although data regarding the possible orientation of the COOH region were not conclusive. The role of the sole cysteine residue, Cys-42, in QacC was studied by using the sulfhydryl reagent N-ethylmaleimide and site-directed mutagenesis. N-Ethylmaleimide was shown to inhibit qacC-mediated ethidium export. Multiple amino acid substitutions were made for Cys-42, and mutations at this location had various effects on resistance specificity. This suggests that the Cys-42 residue may be located near a region of QacC that is involved in substrate recognition. Mutagenesis of conserved residues in QacC indicated that Tyr-59 and Trp-62 also play an essential structural or functional role in QacC.


Perianal Crohn's disease.
The management of perianal Crohn's disease is difficult. A wide variety of treatment options exist although few are evidence based. A search was conducted using the National Library of Medicine for articles on perianal Crohn's disease and its incidence, classification, assessment and management. Perianal Crohn's disease can manifest as skin tags, ulcers, fissures, abscesses, fistulas or stenoses. Improved radiological imaging with endoanal anal ultrasonography and magnetic resonance imaging has improved its assessment and may be used to predict outcome after surgery. Many treatment options exist. During acute complications they are generally aimed at resolving the immediate problem and limiting damage to anal and perianal tissues; this may be a 'bridge' to definitive treatment. The likelihood of success of definitive treatment must be weighed against the risk of complications, especially faecal incontinence.


Trail Blazing or Jam Session? Towards a New Concept of Clinical Decision-making.
Clinical decision-making (CDM) is key in learning to be a doctor as the defining activity in their clinical work. CDM is often portrayed in the literature as similar to 'trail blazing'; the doctor as the core agent, clearing away obstacles on the path towards diagnosis and treatment. However, in a fieldwork of young doctors in Denmark, it was difficult connect their practice to this image. This paper presents the exploration of this discrepancy in the heart of medical practice and how an alternative image emerged; that of a 'jam session'. The exploration is represented as a case-based hypothesis-testing: first, a theoretically and empirically informed hypothesis (H0) of how doctors perform CDM is developed. In H0, CDM is a stepwise process of reasoning about clinical data, often influenced by outside contextual factors. Then, H0 is tested against a case from ethnographic fieldwork with doctors going through internship. Although the case is chosen for characteristics that make it 'most likely' to verify the hypothesis, verification proves difficult. The case challenges preconceptions in CDM literature about chronology, context, objectivity, cognition, agency, and practice. The young doctor is found not to make decisions, but rather to participate in CDM; an activity akin to the dynamics found in a jam session. Their participation circles in and through four concurrent interrelated constructions that suggest a new conceptualization of CDM; a starting point for a deeper understanding of actual practice in a changing clinical environment.


Estradiol impairs response inhibition in young and middle-aged, but not old rats.
Estrogens have been shown to have a strong influence on such cognitive domains as spatial memory, response learning, and several tasks of executive function, including both working memory and attention. However, the effects of estrogens on inhibitory control and timing behavior, both important aspects of executive function, have received relatively little attention. We examined the effects of estradiol on inhibitory control and timing behavior using a differential reinforcement of low rates of responding (DRL) task. Ovariectomized young (3 month), middle-aged (12 month), and old (18 month) Long-Evans rats were implanted with Silastic implants containing 0, 5 or 10% 17β-estradiol in cholesterol vehicle and were tested on a DRL task requiring them to wait 15s between lever presses to receive a food reinforcer. The ratio of reinforced to non-reinforced lever presses did not differ across age in the cholesterol vehicle group. Conversely, 17β-estradiol impaired learning of the DRL task in young and middle-aged rats, but the learning of old rats was not impaired relative to vehicle controls following either 5% or 10% 17β-estradiol treatment. Overall, old rats also made fewer lever presses than both the young and middle-aged rats. These results provide new evidence that estrogens impair inhibitory control, an important aspect of self regulation, and add to existing evidence that estrogens differentially affect cognition at different ages.


Local histograms and image occlusion models.
The local histogram transform of an image is a data cube that consists of the histograms of the pixel values that lie within a fixed neighborhood of any given pixel location. Such transforms are useful in image processing applications such as classification and segmentation, especially when dealing with textures that can be distinguished by the distributions of their pixel intensities and colors. We, in particular, use them to identify and delineate biological tissues found in histology images obtained via digital microscopy. In this paper, we introduce a mathematical formalism that rigorously justifies the use of local histograms for such purposes. We begin by discussing how local histograms can be computed as systems of convolutions. We then introduce probabilistic image models that can emulate textures one routinely encounters in histology images. These models are rooted in the concept of image occlusion. A simple model may, for example, generate textures by randomly speckling opaque blobs of one color on top of blobs of another. Under certain conditions, we show that, on average, the local histograms of such model-generated-textures are convex combinations of more basic distributions. We further provide several methods for creating models that meet these conditions; the textures generated by some of these models resemble those found in histology images. Taken together, these results suggest that histology textures can be analyzed by decomposing their local histograms into more basic components. We conclude with a proof-of-concept segmentation-and-classification algorithm based on these ideas, supported by numerical experimentation.


Verticillium Wilt of Spineless Safflower Caused by Verticillium dahliae in California.
Spineless selections of Carthamus tinctorius (safflower) are grown as commercial field grown cutflower crops in coastal California. In 2010, field plantings of spineless safflower in Santa Clara County developed symptoms of a wilt disease. Affected plants were stunted and slow to develop. As plants developed flower buds, lower leaves turned yellow and wilted. As disease developed, lower leaves turned tan and desiccated; in extreme cases, the entire plant wilted and died. Examination of the taproot and stem vascular tissue revealed a tan to light brown, longitudinal, vascular discoloration. Disease distribution was patchy but in some plantings up to 50% of the plants were unharvestable due to loss of quality or plant death. Isolation from symptomatic vascular tissue consistently resulted in the recovery of a fungus with white aerial mycelium, verticillate conidiophores, single-celled, ovoid to ellipsoid, hyaline conidia, and solitary black microsclerotia that were rounded to elongated or irregular in shape. Three single conidial isolates were chosen for species and race identification after DNA extraction using the FastDNASPIN Kit (MP Biomedicals, Solon, OH). Verticillium species-specific PCR amplified a 500-bp amplicon that is specific to Verticillium dahliae from all three isolates. All three isolates also amplified the race 2-specific 270-bp band in PCR. No amplification was observed in race 1-specific PCR. Based on morphological and molecular data, the fungus was identified as V. dahliae (1,3). Pathogenicity of two isolates was tested individually by soil drench inoculations using 10 ml of conidial suspensions (7 × 106 conidia/ml) for each of 10 containerized plants grown in a peat moss mix in 7.6 cm diameter pots. Five safflower selections were inoculated and maintained in a greenhouse. After 6 weeks, as plants began to form flowers, inoculated plants showed lower leaf dieback and plant wilting. Vascular discoloration was observed when plants were dissected. V. dahliae was consistently recovered from symptomatic tissue. Control plants that only received water did not develop symptoms. The experiment was repeated and the results were consistent. To our knowledge, this is the first report of Verticillium wilt of ornamental, spineless safflower. This disease has been reported previously on agronomic safflower grown as an oilseed crop (2). This finding has significance for coastal crop rotation decisions as ornamental safflower is yet another host that could augment V. dahliae soil inoculum levels for crops such as strawberry and vegetables. References: (1) P. Inderbitzin et al. PLoS One 6: e28341, 2011. (2) J. M. Klisiewicz. Plant Dis. 65:237, 1981. (3) Maruthachalam et al. Phytopathology 100:1222, 2010.


A low cost, safe, disposable, rapid and self-sustainable paper-based platform for diagnostic testing: lab-on-paper.
There is a strong interest in the use of biopolymers in the electronic and biomedical industries, mainly towards low-cost applications. The possibility of developing entirely new kinds of products based on cellulose is of current interest, in order to enhance and to add new functionalities to conventional paper-based products. We present our results towards the development of paper-based microfluidics for molecular diagnostic testing. Paper properties were evaluated and compared to nitrocellulose, the most commonly used material in lateral flow and other rapid tests. Focusing on the use of paper as a substrate for microfluidic applications, through an eco-friendly wax-printing technology, we present three main and distinct colorimetric approaches: (i) enzymatic reactions (glucose detection); (ii) immunoassays (antibodies anti-Leishmania detection); (iii) nucleic acid sequence identification (Mycobacterium tuberculosis complex detection). Colorimetric glucose quantification was achieved through enzymatic reactions performed within specific zones of the paper-based device. The colouration achieved increased with growing glucose concentration and was highly homogeneous, covering all the surface of the paper reaction zones in a 3D sensor format. These devices showed a major advantage when compared to the 2D lateral flow glucose sensors, where some carryover of the coloured products usually occurs. The detection of anti-Leishmania antibodies in canine sera was conceptually achieved using a paper-based 96-well enzyme-linked immunosorbent assay format. However, optimization is still needed for this test, regarding the efficiency of the immobilization of antigens on the cellulose fibres. The detection of Mycobacterium tuberculosis nucleic acids integrated with a non-cross-linking gold nanoprobe detection scheme was also achieved in a wax-printed 384-well paper-based microplate, by the hybridization with a species-specific probe. The obtained results with the above-mentioned proof-of-concept sensors are thus promising towards the future development of simple and cost-effective paper-based diagnostic devices.


Physiologically informed dynamic causal modeling of fMRI data.
The functional MRI (fMRI) signal is an indirect measure of neuronal activity. In order to deconvolve the neuronal activity from the experimental fMRI data, biophysical generative models have been proposed describing the link between neuronal activity and the cerebral blood flow (the neurovascular coupling), and further the hemodynamic response and the BOLD signal equation. These generative models have been employed both for single brain area deconvolution and to infer effective connectivity in networks of multiple brain areas. In the current paper, we introduce a new fMRI model inspired by experimental observations about the physiological underpinnings of the BOLD signal and compare it with the generative models currently used in dynamic causal modeling (DCM), a widely used framework to study effective connectivity in the brain. We consider three fundamental aspects of such generative models for fMRI: (i) an adaptive two-state neuronal model that accounts for a wide repertoire of neuronal responses during and after stimulation; (ii) feedforward neurovascular coupling that links neuronal activity to blood flow; and (iii) a balloon model that can account for vascular uncoupling between the blood flow and the blood volume. Finally, we adjust the parameterization of the BOLD signal equation for different magnetic field strengths. This paper focuses on the form, motivation and phenomenology of DCMs for fMRI and the characteristics of the various models are demonstrated using simulations. These simulations emphasize a more accurate modeling of the transient BOLD responses - such as adaptive decreases to sustained inputs during stimulation and the post-stimulus undershoot. In addition, we demonstrate using experimental data that it is necessary to take into account both neuronal and vascular transients to accurately model the signal dynamics of fMRI data. By refining the models of the transient responses, we provide a more informed perspective on the underlying neuronal process and offer new ways of inferring changes in local neuronal activity and effective connectivity from fMRI.


Voxel-based analysis of grey and white matter degeneration in cervical spondylotic myelopathy.
In this prospective study, we made an unbiased voxel-based analysis to investigate above-stenosis spinal degeneration and its relation to impairment in patients with cervical spondylotic myelopathy (CSM). Twenty patients and 18 controls were assessed with high-resolution MRI protocols above the level of stenosis. Cross-sectional areas of grey matter (GM), white matter (WM), and posterior columns (PC) were measured to determine atrophy. Diffusion indices assessed tract-specific integrity of PC and lateral corticospinal tracts (CST). Regression analysis was used to reveal relationships between MRI measures and clinical impairment. Patients showed mainly sensory impairment. Atrophy was prominent within the cervical WM (13.9%, p = 0.004), GM (7.2%, p = 0.043), and PC (16.1%, p = 0.005). Fractional anisotropy (FA) was reduced in the PC (-11.98%, p = 0.006) and lateral CST (-12.96%, p = 0.014). In addition, radial (+28.47%, p = 0.014), axial (+14.72%, p = 0.005), and mean (+16.50%, p = 0.001) diffusivities were increased in the PC. Light-touch score was associated with atrophy (R(2) = 0.3559, p = 0.020) and FA (z score 3.74, p = 0.003) in the PC, as was functional independence and FA in the lateral CST (z score 3.68, p = 0.020). This study demonstrates voxel-based degeneration far above the stenosis at a level not directly affected by the compression and provides unbiased readouts of tract-specific changes that relate to impairment.


Renal failure induces atrial arrhythmogenesis from discrepant electrophysiological remodeling and calcium regulation in pulmonary veins, sinoatrial node, and atria.
Renal failure (RF) increases the risk of atrial fibrillation (AF), but arrhythmogenic mechanism is unclear. The present study investigated the electrophysiological effects of RF on AF trigger (pulmonary veins, PVs) and substrate (atria) and evaluated potential underlying mechanisms. Electrocardiographic, echocardiographic, and biochemical studies were conducted in rabbits with and without antibiotic-induced mild (creatinine=1.5-6.0 mg/dl) and advanced (creatinine>6.0 mg/dl) RF. Conventional microelectrode techniques, western blotting, and histological examinations were performed using the isolated rabbit PV, left atrium (LA), right atrium (RA) and sinoatrial node (SAN). Advanced RF rabbits (n=18) had a higher incidence (33.3% vs. 11.1% and 0%, p<0.05) of atrial arrhythmia than mild RF (n=18) and control (n=18) rabbits. Advanced RF rabbits exhibited faster PV spontaneous activities, longer action potential duration (APD) in the LA, higher fibrosis in the LA, and slower SAN beating rates than control rabbits, but had a similar APD and fibrosis in the RA. Caffeine (1 mM) increased advanced RF PV arrhythmogenesis, which is blocked by flecainide (10 μM), or KB-R7943 (10 μM). Moreover, advanced RF rabbits had a higher expression of the Na+/Ca2+ exchanger, protein kinase A, phosphorylated ryanodine receptor (Serine 2808), and phosphorylated phospholamban (Serine 16) in PVs, and a higher expression of Cav 1.2 in the LA, and a lower expression of hyperpolarization-activated cyclic nucleotide-gated potassium channel 4 in the SAN. Advanced RF increases atrial arrhythmia by modulating the distinctive electrophysiological characteristics of the PV, LA, and SAN.


CVID-Associated Tumors: Czech Nationwide Study Focused on Epidemiology, Immunology, and Genetic Background in a Cohort of Patients With CVID.
Background: Common variable immunodeficiency disorder (CVID) is one of the most frequent inborn errors of immunity, increased occurrence of malignancies, particularly lymphomas, and gastric cancers, has long been noted among CVID patients. Multifactorial etiology, including immune dysregulation, infections, chronic inflammation, or genetic background, is suggested to contribute to tumor development. Here, we present the results of the first Czech nationwide study focused on epidemiology, immunology and genetic background in a cohort of CVID patients who also developed tumors Methods: The cohort consisted of 295 CVID patients followed for 3,070 patient/years. Standardized incidence ratio (SIR) was calculated to determine the risk of cancer, and Risk ratio (RR) was established to evaluate the significance of comorbidities. Moreover, immunophenotyping, including immunoglobulin levels and lymphocyte populations, was assessed. Finally, Whole exome sequencing (WES) was performed in all patients with lymphoma to investigate the genetic background. Results: Twenty-five malignancies were diagnosed in 22 patients in a cohort of 295 CVID patients. SIR was more than 6 times greater in comparison to the general population. The most common neoplasias were gastric cancers and lymphomas. History of Immune thrombocytopenic purpura (ITP) was established as a potential risk factor, with over 3 times higher risk of cancer development. The B cell count at diagnosis of lymphoma was reduced in the lymphoma group; moreover, post-treatment B and T cell lymphopenia, associated with poorer outcome, was found in a majority of the patients. Intriguingly, no NK cell depression was observed after the chemotherapy. WES revealed heterogeneous genetic background among CVID patients with tumors, identifying gene variants associated with primary immunodeficiencies (such as CTLA4, PIK3CD, PMS2) and/or increased cancer susceptibility (including BRCA1, RABEP1, EP300, KDM5A). Conclusions: The incidence of malignancy in our CVID cohort was found to be more than 6 times greater compared to the general population. Gastric cancers and lymphomas were the most frequently diagnosed tumors. ITP was identified as a risk factor for malignancy in CVID patients. WES analysis confirmed a wide genetic heterogeneity among CVID patients. The identified causative or modifying gene variants pointed to errors in mechanisms contributing to both immunodeficiency and malignancy.


The light-from-above prior is intact in autistic children.
Sensory information is inherently ambiguous. The brain disambiguates this information by anticipating or predicting the sensory environment based on prior knowledge. Pellicano and Burr (2012) proposed that this process may be atypical in autism and that internal assumptions, or "priors," may be underweighted or less used than in typical individuals. A robust internal assumption used by adults is the "light-from-above" prior, a bias to interpret ambiguous shading patterns as if formed by a light source located above (and slightly to the left) of the scene. We investigated whether autistic children (n=18) use this prior to the same degree as typical children of similar age and intellectual ability (n=18). Children were asked to judge the shape (concave or convex) of a shaded hexagon stimulus presented in 24 rotations. We estimated the relation between the proportion of convex judgments and stimulus orientation for each child and calculated the light source location most consistent with those judgments. Children behaved similarly to adults in this task, preferring to assume that the light source was from above left, when other interpretations were compatible with the shading evidence. Autistic and typical children used prior assumptions to the same extent to make sense of shading patterns. Future research should examine whether this prior is as adaptable (i.e., modifiable with training) in autistic children as it is in typical adults.


TRH (protirelin) in depressed alcoholic men. Behavioral changes and endocrine responses.
Chronic alcoholics with secondary depression were treated with protirelin in a double-blind, placebo-controlled study. Behavioral data, collected only during the acute alcohol withdrawal state, indicated a beneficial effect of protirelin three hours after injection, but not during subsequent days. Injections caused only mild and infrequent subjective side effects and no cardiovascular effects. Endocrine data were recorded in the acute withdrawal state and after clinical remission. Findings in the acute state suggested thyroid activation and increased central dopaminergic activity, as evidenced by elevated baseline levels of growth hormone, low baseline levels of prolactin, and blunted thyroid-stimulating hormone (TSH) response to protirelin. The first two abnormalities returned to normal levels in the remission state. A blunted TSH response was observed in both the acute and the remission states. Partial persistence of this finding suggests that TSH blunting may not be solely state-dependent. In the acute withdrawal state, TSH blunting was associated with favorable behavioral responses to protirelin.


Ruthenium water oxidation catalysts containing the non-planar tetradentate ligand, biisoquinoline dicarboxylic acid (biqaH2).
Two ruthenium complexes containing the tetradentate ligand [1,1'-biisoquinoline]-3,3'-dicarboxylic acid, and 4-picoline or 6-bromoisoquinoline as axial ligands have been prepared. The complexes have been fully characterised and initial studies on their potential to function as molecular water oxidation catalysts have been performed. Both complexes catalyse the oxidation of water in acidic media with CeIV as a stoichiometric chemical oxidant, although turnover numbers and turnover frequencies are modest when compared with the closely related Ru-bda and Ru-pda analogues. Barriers for the water nucleophilic attack and intermolecular coupling pathways were obtained from density functional theory calculations and the crucial influence of the ligand framework in determining the most favourable reaction pathway was elucidated from a combined analysis of the theoretical and experimental results.


Feasibility of treating prehypertension with an angiotensin-receptor blocker.
Prehypertension is considered a precursor of stage 1 hypertension and a predictor of excessive cardiovascular risk. We investigated whether pharmacologic treatment of prehypertension prevents or postpones stage 1 hypertension. Participants with repeated measurements of systolic pressure of 130 to 139 mm Hg and diastolic pressure of 89 mm Hg or lower, or systolic pressure of 139 mm Hg or lower and diastolic pressure of 85 to 89 mm Hg, were randomly assigned to receive two years of candesartan (Atacand, AstraZeneca) or placebo, followed by two years of placebo for all. When a participant reached the study end point of stage 1 hypertension, treatment with antihypertensive agents was initiated. Both the candesartan group and the placebo group were instructed to make changes in lifestyle to reduce blood pressure throughout the trial. A total of 409 participants were randomly assigned to candesartan, and 400 to placebo. Data on 772 participants (391 in the candesartan group and 381 in the placebo group; mean age, 48.5 years; 59.6 percent men) were available for analysis. During the first two years, hypertension developed in 154 participants in the placebo group and 53 of those in the candesartan group (relative risk reduction, 66.3 percent; P<0.001). After four years, hypertension had developed in 240 participants in the placebo group and 208 of those in the candesartan group (relative risk reduction, 15.6 percent; P<0.007). Serious adverse events occurred in 3.5 percent of the participants assigned to candesartan and 5.9 percent of those receiving placebo. Over a period of four years, stage 1 hypertension developed in nearly two thirds of patients with untreated prehypertension (the placebo group). Treatment of prehypertension with candesartan appeared to be well tolerated and reduced the risk of incident hypertension during the study period. Thus, treatment of prehypertension appears to be feasible. (ClinicalTrials.gov number, NCT00227318.).


Retention loss of resin based fissure sealants - a valid predictor for clinical outcome?
The null-hypothesis that retention loss of resin fissure sealants predicts caries manifestation no more accurately than random values was tested. Systematic reviews were checked and electronic databases searched for clinical trials. Trials reporting on the retention of resin sealants and caries occurrence in permanent molar teeth, with minimum 24-month follow-up period, were included. Extracted data: number of sealed teeth, number of teeth without completely retained sealants, number of sealed teeth with caries. The number of teeth with complete sealant retention and absence of carious lesions/cavities was calculated; the predictive outcomes: true/false positive; false/true negative were established. Random values were generated as control-data. Diagnostic Odds ratios (DOR) were computed and tested for statistical difference. Summary Receiver Operating Characteristic curves were plotted. 95 trials were found. Median DOR values were 1.21 and 0.28 for test- and control data, respectively. Wilcoxon test (z = 0.56; p = 0.58) and Sign test (z = 1.38; p = 0.17) results were statistically non-significant. The null-hypothesis was not rejected. Predictions based on the retention loss of resin sealants, regarding caries manifestation, was no more accurate than random guesses. Sealant retention loss appears not to be a valid predictor for clinical outcome.


Reduced mRNA abundance of the main enzymes involved in methionine metabolism in human liver cirrhosis and hepatocellular carcinoma.
It has been known for at least 50 years that alterations in methionine metabolism occur in human liver cirrhosis. However, the molecular basis of this alteration is not completely understood. In order to gain more insight into the mechanisms behind this condition, mRNA levels of methionine adenosyltransferase (MAT1A), glycine methyltransferase (GNMT), methionine synthase (MS), betaine homocysteine methyltransferase (BHMT) and cystathionine beta-synthase (CBS) were examined in 26 cirrhotic livers, five hepatocellular carcinoma (HCC) tissues and ten control livers. The expression of the above-mentioned genes was determined by quantitative RT-PCR analysis. Methylation of MAT1A promoter was assessed by methylation-sensitive restriction enzyme digestion of genomic DNA. When compared to normal livers MAT1A, GNMT, BHMT, CBS and MS mRNA contents were significantly reduced in liver cirrhosis. Interestingly, MAT1A promoter was hypermethylated in the cirrhotic liver. HCC tissues also showed decreased mRNA levels of these enzymes. These findings establish that the abundance of the mRNA of the main genes involved in methionine metabolism is markedly reduced in human cirrhosis and HCC. Hypermethylation of MAT1A promoter could participate in its reduced expression in cirrhosis. These observations help to explain the hypermethioninemia, hyperhomocysteinemia and reduced hepatic glutathione content observed in cirrhosis.


Correlation between CYP1A1 polymorphisms and susceptibility to glyphosate-induced reduction of serum cholinesterase: A case-control study of a Chinese population.
Glyphosate (GLP) is one of the most common herbicides worldwide. The serum cholinesterase (ChE) may be affected when exposed to glyphosate. Reduction of serum ChE by herbicides is probably related to cytochrome P450 (CYP450) family polymorphisms. We suspect that the abnormal ChE caused by GLP could be correlated with the CYP family members. To determine whether CYP1B1 (rs1056827 and rs1056836) and CYP1A1 (rs1048943) gene polymorphisms and individual susceptibility to GLP-induced ChE abnormalities were interrelated in the Chinese Han population, we performed this genetic association study on a total of 230 workers previously exposed to GLP, including 115 cases with reduced serum ChE and 115 controls with normal serum ChE. Two even groups of cases and controls were enrolled. The CYP1A1 and CYP1B1 polymorphisms in both groups were genotyped using TaqMan. Subjects with the CYP1A1 rs619586 genotypes showed an increased risk of GLP-induced reduction of serum ChE, which was more evident in the following subgroups: female, > 35 years old, history of GLP exposure time <10 years and >10 years, nonsmoker and nondrinker. The results show that CYP1A1 rs619586 was significantly associated with the GLP-induced reduction in serum ChE and could be a biomarker of susceptibility for Chinese GLP exposed workers. Because of a large number of people exposed to glyphosate, this study has a significance in protecting their health.


Autologous serum for ocular surface diseases.
Autologous serum has been used to treat dry eye syndrome for many years. It contains several growth factors, vitamins, fibronectin and other components that have been considered important for corneal and conjunctival integrity. Serum eye drops are usually prepared as an unpreserved blood solution. The serum is by nature well tolerated and its biochemical properties are somewhat similar to natural tears. Autologous serum eye drops have been reported to be effective for the treatment of severe dry eye-related ocular surface disorders (Sjögren's syndrome), and also other entities such as superior limbic keratoconjunctivitis, graft-versus-host disease, Stevens-Johnson syndrome, ocular cicatricial pemphigoid, recurrent or persistent corneal erosions, neurotrophic keratopathy, Mooren's ulcer, aniridic keratopathy, filtering blebs after trabeculectomy, and post-keratorefractive surgery. The purpose of this study is to review the recently published literature on ocular surface diseases treated with human autologous serum eye drops.


Preparation and characterization of directly compactible layer-by-layer nanocoated cellulose.
Microcrystalline cellulose is a commonly used direct compression tablet diluent and binder. It is derived from purified α-cellulose in an environmentally unfriendly process that involves mineral acid catalysed hydrolysis. In this study Kraft softwood fibers was nanocoated using a layer-by-layer self-assembling process. Powder flow and compactibility results showed that the application of nano-thin polymer layers on the fibers turned non-flowing, non-compacting cellulose into powders that can be used in the direct compression of tablets. The powder flow properties and tableting indices of compacts compressed from these nanocoated microfibers were similar or better than that of directly compactible microcrystalline cellulose powders. Cellulose microfibers coated with four PSS/PVP bilayers had the best compaction properties while still producing tablets that were able to absorb water and disintegrate and did not retard the dissolution of a model drug acetaminophen. The advantages of nanocoating rather than traditional pharmaceutical coating are that it add less than 1% to the weight of the fibers and allows control of the molecular properties of the surface and the thickness of the coat to within a few nanometers. This process is potentially friendlier to the environment because of the type and quantity of materials used. Also, it does not involve acid-catalyzed hydrolysis and neutralization of depolymerized cellulose.


IL-8 -251A/T polymorphism contributes to coronary artery disease susceptibility in a Chinese population.
Interleukin-8 (IL-8) is a mediator of inflammation and plays an important role in regulating immune responses. To date, several studies have tested the association between IL-8 gene polymorphisms and development of coronary artery disease (CAD), but their results have proved to be inconsistent. We conducted an investigation to assess the relationship between the IL-8 -251A/T (rs4073) sequence variant and CAD in a Chinese population. Between April 2013 and January 2015, 217 patients with coronary angiography-confirmed CAD were enrolled in our study, along with 245 control subjects. IL-8 -251A/T genotyping was performed using a polymerase chain reaction-restriction fragment length polymorphism assay. A chi-square test revealed that IL-8 -251A/T genotype distributions significantly differed between CAD patients and control subjects (chi-square = 8.29, P < 0.02). Moreover, multiple-logistic regression analysis showed that individuals carrying TA [odds ratio (OR) = 1.59, 95% confidence interval (CI) = 1.01-2.57] and AA (OR = 2.06, 95%CI = 1.21-3.52) genotypes were at increased risk of CAD compared to those with the TT genotype. Under dominant (OR = 1.75, 95%CI = 1.13-2.73) and recessive (OR = 1.54, 95%CI = 1.02-2.37) genetic models, the IL-8 -251A/T polymorphism also significantly correlated with CAD. In conclusion, our results suggest that this variant is an independent risk factor for CAD development under codominant, dominant, and recessive models.


The effects of topical antibiotics on eradication and acquisition of third-generation cephalosporin and carbapenem-resistant Gram-negative bacteria in ICU patients; a post hoc analysis from a multicentre cluster-randomized trial.
The aim was to quantify the effects of selective digestive tract decontamination (SDD) consisting of a mouth paste and gastro-enteral suspension, selective oropharyngeal decontamination with a mouth paste (SOD) and 1-2% chlorhexidine (CHX) mouthwash on eradication and acquisition of carriage of third-generation cephalosporin-resistant Enterobacterales (3GCR-E) and carbapenem-resistant Gram-negative bacteria (CR-GNB) in Intensive Care Unit (ICU) patients. This was a nested cohort study within a cluster-randomized cross-over trial in six European countries and 13 ICUs with 8665 patients. Eradication and acquisition during ICU stay of 3GCR-E and CR-GNB were investigated separately in the rectum and respiratory tract for the three interventions and compared with standard care (SC) using Cox-regression competing events analyses. Adjusted cause specific hazard ratios (CSHR) for eradication of rectal carriage for SDD were 1.76 (95% CI 1.31-2.36) for 3GCR-E and 3.17 (95% CI 1.60-6.29) for CR-GNB compared with SC. For the respiratory tract, adjusted CSHR for eradication of 3GCR-E were 1.47 (0.98-2.20) for SDD and 1.38 (0.92-2.06) for SOD compared with SC, and for eradication of CR-GNB these were 0.77 (0.41- 1.45) for SDD and 0.81 (0.44-1.51) for SOD, compared with SC. Adjusted CSHRs for acquisition of rectal carriage during SDD (compared with SC) were 0.51 (0.40-0.64) for 3GCR-E and of 0.56 (0.40-0.78) for CR-GNB. Adjusted CSHRs for acquiring respiratory tract carriage with 3GCR-E compared with SC were 0.38 (0.28-0.50) for SDD and 0.55 (0.42-0.71) for SOD, and for CR-GNB 0.46 (0.33-0.64) during SDD and 0.60 (0.44-0.81) during SOD, respectively. SOD was not associated with eradication or acquisition of 3GCR-E and CR-GNB in the rectum. Among mechanically ventilated ICU patients, SDD was associated with more eradication and less acquisition of 3GCR-E and CR-GNB in the rectum than SC. SDD and SOD were associated with less acquisition of both 3GCR-E and CR-GNB than SC in the respiratory tract.


Sensing of human plasma fibrinogen on polished, chemically etched and carbon treated titanium surfaces by diffractive optical element based sensor.
Adsorption of human plasma fibrinogen (HPF) on 6 differently treated titanium samples (polished, polished and etched, and 4 titanium carbide coatings samples produced by using plasma-enhanced chemical vapour deposition (PECVD) method) is investigated by using diffractive optical element (DOE) sensor. Permittivity (susceptibility) change and fluctuation in optical roughness (R(opt)) of treated titanium surface in the presence of background electrolyte without and with HPF molecules are sensed by using DOE sensor and optical ellipsometry. Correlation between transmitted light and thickness of molecule layer was found. The findings allow to sense temporal organization and severity of adsorption of nano-scale HPF molecules on polished, on polished and etched, and on titanium carbide surface.


Porphyromonas gingivalis stimulates TACE production by T cells.
Tumour necrosis factor-alpha converting enzyme (TACE), also known as ADAM17, is a membrane-bound metalloprotease and disintegrin. It is produced by a number of host cells and is known to shed and release cell-bound cytokines, particularly members of the tumour necrosis factor family. The aim of this study was to investigate the effect of Porphyromonas gingivalis on TACE production by a human T-cell line, to identify putative virulence factors involved in this process, and to investigate the effect of doxycycline. P. gingivalis 6-day culture supernatants were used to challenge Jurkat T cells for 6 h. Secreted and cell-associated TACE levels were measured by enzyme-linked immunosorbent assay, whereas messenger RNA expression was investigated by quantitative real-time polymerase chain reaction. To investigate the involvement of cysteine proteases or proteinaceous components in general, P. gingivalis culture supernatants were treated with the specific chemical inhibitor TLCK or heat-inactivated, respectively. The effect of doxycycline on the regulation of TACE secretion by P. gingivalis was also investigated. P. gingivalis challenge resulted in a concentration-dependent enhancement of TACE messenger RNA expression and protein release by Jurkat cells. TLCK treatment or heat treatment of P. gingivalis culture supernatants decreased TACE release to control levels. Doxycycline inhibited TACE secretion dose dependently. The induction of TACE by T cells in response to P. gingivalis may in turn favour the shedding of host cell-bound cytokines into the local microenvironment, potentially amplifying the inflammatory response. In the present experimental system, P. gingivalis cysteine proteases are involved in TACE release by T cells.


Database for CO2 Separation Performances of MOFs Based on Computational Materials Screening.
Metal-organic frameworks (MOFs) are potential adsorbents for CO2 capture. Because thousands of MOFs exist, computational studies become very useful in identifying the top performing materials for target applications in a time-effective manner. In this study, molecular simulations were performed to screen the MOF database to identify the best materials for CO2 separation from flue gas (CO2/N2) and landfill gas (CO2/CH4) under realistic operating conditions. We validated the accuracy of our computational approach by comparing the simulation results for the CO2 uptakes, CO2/N2 and CO2/CH4 selectivities of various types of MOFs with the available experimental data. Binary CO2/N2 and CO2/CH4 mixture adsorption data were then calculated for the entire MOF database. These data were then used to predict selectivity, working capacity, regenerability, and separation potential of MOFs. The top performing MOF adsorbents that can separate CO2/N2 and CO2/CH4 with high performance were identified. Molecular simulations for the adsorption of a ternary CO2/N2/CH4 mixture were performed for these top materials to provide a more realistic performance assessment of MOF adsorbents. The structure-performance analysis showed that MOFs with Δ Qst0 > 30 kJ/mol, 3.8 Å < pore-limiting diameter < 5 Å, 5 Å < largest cavity diameter < 7.5 Å, 0.5 < ϕ < 0.75, surface area < 1000 m2/g, and ρ > 1 g/cm3 are the best candidates for selective separation of CO2 from flue gas and landfill gas. This information will be very useful to design novel MOFs exhibiting high CO2 separation potentials. Finally, an online, freely accessible database https://cosmoserc.ku.edu.tr was established, for the first time in the literature, which reports all of the computed adsorbent metrics of 3816 MOFs for CO2/N2, CO2/CH4, and CO2/N2/CH4 separations in addition to various structural properties of MOFs.


The human microbiota: the rise of an "empire".
The human body hosts rich and diverse microbial communities. Our microbiota affects the normal human physiology, and compositional changes might alter host homeostasis and, therefore, disease risk. The microbial community structure may sometimes occupy discrete configurations and under certain circumstances vary continuously. The ability to characterize accurately the ecology of human-associated microbial communities became possible by advances in deep sequencing and bioinformatics analyses.


Regional cerebellar volumetric correlates of manual motor and cognitive function.
Cerebellar volume declines with aging. Few studies have investigated age differences in regional cerebellar volume (RCV) and their association with motor and cognitive function. In 213 healthy older adults, we investigated the association of age with motor skills, cognition and RCV. Subsequently, we studied the association of RCV with motor skills and cognition. RCVs were derived from T1-weighted MRI scans using the automated SUIT segmentation method and clustered using principal component analysis (PCA). Motor skill (manual dexterity, tapping speed, bimanual visuomotor coordination, grip force) and cognition (mental rotation, verbal memory, inhibition, mental flexibility) were assessed. Behavioral measures were clustered into compounds using PCA: left hand motor skill, right hand motor skill, verbal memory and mental flexibility, and mental rotation & inhibition. Volume of the rostral middle frontal gyri (rMFG) and premotor areas (PMA) were related to performance for reference. Analyses were adjusted for age, sex, and education. Volume of the cerebellar anterior lobe and top of the superior posterior lobe were positively associated with motor skill. Volume of the bottom part of the posterior superior lobe and the inferior posterior lobe was positively associated with cognition. PMA volume was associated with cognition and motor skill and rMFG volume with motor skill. Although these results did not survive FDR correction, their effect sizes suggest that regional cerebellar volume selectively contributes to cognitive and motor skill. Effect sizes of cerebellar associations with performance were similar to those of rMFG/PMA and performance suggesting parallel contributions to performance.


Drop weld thermal injuries to the middle ear.
Drop weld injuries to the tympanic membrane and middle ear caused by hot sparks or molten slag are a rare but significant injury. Steel workers and welders who are regularly exposed to flying sparks and molten metal slag are predisposed. This type of transtympanic thermal injury occurs when the slag literally drops into the external auditory canal and burns through the tympanic membrane. A spectrum of severity of injury occurs which includes chronic tympanic membrane perforation, chronic otorrhoea, facial nerve injury and deafness. Chronic tympanic membrane perforation is the most common sequelae and is perhaps one of the most challenging of all perforations to repair The combination of direct thermal injury and foreign body reaction results in continuing or recurrent suppuration. The foreign body reaction is due to the embedding of metal slag in the promontorial mucosa. We present a case of drop weld injury to the left tympanic membrane, resulting in chronic middle ear inflammation, otorrhoea and tympanic perforation. CAT scan clearly demonstrated a metallic promontorial foreign body with localised bone erosion. We emphasise the importance of removing these foreign bodies and recommend a cartilage reinforced underlay tympanoplasty technique to repair these perforations. Transtympanic thermal trauma is a preventable occupational injury, which is best, avoided by earplugs and increased awareness.


XMAP215 is a long thin molecule that does not increase microtubule stiffness.
XMAP215 is a microtubule associated protein that speeds microtubule plus end growth by seven- to tenfold and protects these ends from destabilization by the Kin I kinesin, XKCM1. To understand the mechanisms responsible for these activities, it is necessary to know the structure of XMAP215. By unidirectional shadowing and electron microscopy, XMAP215 appeared as an elongate molecule of 60+/-18 nm, suggesting that XMAP215 could span up to seven to eight tubulin dimers along a protofilament. Most XMAP215 molecules were straight but a subset were bent suggesting that XMAP215 is flexible. Antibodies to the C terminus labeled one end of XMAP215 with no evidence for XMAP215 dimerization. Incubation of XMAP215 and tubulin at 4 degrees C resulted in assembly of curved protofilaments, which appeared to be incomplete tubulin rings. Measurements from rotary shadowed samples showed that tubulin/XMAP215 partial rings had an average width of 8.8+/-1.8 nm compared with 5.6+/-1.1 nm for rings assembled from tubulin dimers alone, suggesting that XMAP215 adds a width of approximately 3.2 nm to the curved tubulin protofilament. XMAP215 did not change the radius of curvature of these partial tubulin rings. Measurements of microtubule flexural rigidity by thermal fluctuations showed that XMAP215 did not change microtubule rigidity. Finally, sequence analysis shows that the N-terminal half of XMAP215 contains four repeats, each composed of multiple HEAT repeats.


Glycation of low-density lipoproteins by methylglyoxal and glycolaldehyde gives rise to the in vitro formation of lipid-laden cells.
Previous studies have implicated the glycoxidative modification of low-density lipoprotein (LDL) by glucose and aldehydes (apparently comprising both glycation and oxidation), as a causative factor in the elevated levels of atherosclerosis observed in diabetic patients. Such LDL modification can result in unregulated cellular accumulation of lipids. In previous studies we have characterized the formation of glycated, but nonoxidized, LDL by glucose and aldehydes; in this study we examine whether glycation of LDL, in the absence of oxidation, gives rise to lipid accumulation in arterial wall cell types. Glycated LDLs were incubated with macrophage, smooth muscle, or endothelial cells. Lipid loading was assessed by HPLC analysis of cholesterol and individual esters. Oxidation was assessed by cholesterol ester loss and 7-ketocholesterol formation. Cell viability was assessed by lactate dehydrogenase release and cell protein levels. Glycation of LDL by glycolaldehyde and methylglyoxal, but not glucose (in either the presence or absence of copper ions), resulted in cholesterol and cholesterol ester accumulation in macrophage cells, but not smooth muscle or endothelial cells. The extent of lipid accumulation depends on the degree of glycation, with increasing aldehyde concentration or incubation time, giving rise to greater extents of particle modification and lipid accumulation. Modification of lysine residues appears to be a key determinant of cellular uptake. These results are consistent with LDL glycation, in the absence of oxidation, being sufficient for rapid lipid accumulation by macrophage cells. Aldehyde-mediated "carbonyl-stress" may therefore facilitate the formation of lipid-laden (foam) cells in the artery wall.


Neural excitation of the larval firefly photocyte: slow depolarization possibly mediated by a cyclic nucleotide.
1. In firefly larvae, extracellular recordings from the light organ nerve show that a volley of action potentials elicits a glow of an intact animal. 2. Intracellular recordings from the photocytes show that they respond to nerve stimulation with a slow, graded depolarization which precedes light emission. The depolarization begins about 0-5 s after the nerve is stimulated; it peaks about 1 s after stimulation; and subsides about 2-5 s after the stimulus. The glow increases fastest when the photocyte depolarization is at its peak and lasts 5-15 s. 3. Photocyte depolarization is associated with a decrease in the input resistance of the cell. 4. Adrenergic receptors in the light organ are pharmacologically similar to vertebrate alpha-receptors. 5. Phophodiesterase inhbitors, aminophylline and theophylline, cause the light organ to glow, suggesting that cyclic nucleotides may mediate the effect of the adrenergic nerve transmitter.


Formation of virus-like particles when the polyprotein gene (segment A) of infectious bursal disease virus is expressed in insect cells.
The baculovirus expression vector system was used to examine the expression of the full-length infectious bursal disease virus (IBDV) segment A cDNA, which encodes the structural proteins in a polyprotein precursor that is autocatalytically cleaved to VPX, VP3, and VP4. No VP2 was observed in lysates of recombinant baculovirus infected cells indicating the lack of processing of VPX to VP2 in this system. Virus-like particles (VLP) were purified from the infected insect cells, and on negative staining electron microscopy, looked very similar to authentic IBDV particles in shape and size, suggesting that processing of VPX to VP2 is not necessary for capsid assembly.


Suppression of Pythium and Phytophthora Damping-Off of Rooibos by Compost and a Combination of Compost and Nonpathogenic Pythium Taxa.
Pathogenic oomycetes, including Phytophthora cinnamomi and several Pythium spp. (Pythium irregulare, P. mamillatum, P. myriotylum, and P. pyrilobum), cause serious damping-off problems in rooibos (Aspalathus linearis) nurseries. The management of these pathogens in organic nurseries is problematic, because phenylamide fungicides may not be used. Compost, or compost in combination with Pythium taxa that are nonpathogenic to rooibos (P. acanthicum, P. cederbergense, and Pythium RB II), were investigated as alternative management options. Compost was able to suppress damping-off caused by several oomycete isolates but there was within- and between-species variation among the 30 evaluated isolates. This phenomenon was observed using two compost batches (A and B) sourced from independent suppliers. Compost B significantly reduced damping-off caused by 60% of the isolates, whereas compost A controlled only 37% of the isolates. The pathogens that were more readily controlled by both composts included P. mamillatum and P. pyrilobum, whereas the composts were ineffective at suppressing damping-off caused by >62% of P. irregulare and >50% of P. myriotylum isolates. Based on the evaluation of one Phytophthora cinnamomi isolate, this pathogen may also be controlled by compost. Neither of the composts as a stand-alone treatment could suppress damping-off caused by a combination of pathogenic species (P. cinnamomi, Pythium irregulare, P. mamillatum, P. myriotylum, and P. pyrilobum). However, damping-off was significantly reduced when nonpathogenic Pythium taxa (P. acanthicum, P. cederbergense, and Pythium RB II) were combined with the composts. Similarly, damping-off caused by a P. irregulare isolate that was not suppressed by either of the composts alone was significantly suppressed when the two composts were inoculated with the nonpathogenic Pythium taxa.


Disulfide bond engineered into T4 lysozyme: stabilization of the protein toward thermal inactivation.
By recombinant DNA techniques, a disulfide bond was introduced at a specific site in T4 lysozyme, a disulfide-free enzyme. This derivative retained full enzymatic activity and was more stable toward thermal inactivation than the wild-type protein. The derivative, T4 lysozyme (Ile3----Cys), was prepared by substituting a Cys codon for an Ile codon at position 3 in the cloned lysozyme gene by means of oligonucleotide-dependent, site-directed mutagenesis. The new gene was expressed in Escherichia coli under control of the (trp-lac) hybrid tac promoter, and the protein was purified. Mild oxidation generated a disulfide bond between the new Cys3 and Cys97, one of the two unpaired cysteines of the native molecule. Oxidized T4 lysozyme (Ile3----Cys) exhibited specific activity identical to that of the wild-type enzyme when measured at 20 degrees C in a cell-clearing assay. The cross-linked protein was more stable than the wild type during incubation at elevated temperatures as determined by recovered enzymatic activity at 20 degrees C.


Properties and distribution of a lectin-like hemagglutinin differentially expressed by murine stromal tissue macrophages.
We describe a novel hemagglutinin which is differentially expressed on murine stromal tissue macrophages. Resident bone marrow macrophages (RBMM), which are physically associated with immature, proliferating hematopoietic cells in vivo, formed striking rosettes with unopsonized sheep erythrocytes (E) in vitro, unlike resident peritoneal macrophages (RPM). Binding of E was macrophage (M phi) specific, not accompanied by ingestion and independent of temperature (0-37 degrees C), divalent cations, and the metabolic inhibitors azide and iodoacetate. Pretreatment of RBMM with trypsin prevented rosette formation, but neuraminidase enhanced it. Conversely, binding was virtually abrogated if E were pretreated with neuraminidase, whereas trypsin pretreatment of the ligand resulted in a slight enhancement. The lectin-like nature of the E receptor (SER), with specificity for sialylated glycoconjugates, was consistent with the inhibition of binding we saw with neuraminyllactose or the ganglioside GD1a (50% inhibition at 5-10 mM and 11 microM, respectively). Expression of SER on freshly isolated RBMM was heterogeneous and exhibited a striking inverse correlation with expression of Ia antigens. During cultivation in 10% FCS, levels of SER on RBMM declined with a half-life of approximately 24 h. Other cell surface changes induced by cultivation included a transient increase in expression of Ia antigen and acquisition of Mac-1. To determine whether SER was expressed on other stromal M phi populations, adherent cells were isolated from various tissues by collagenase digestion or lavage. Binding of E was highest on RBMM and lymph node stromal M phi, at intermediate levels on Kupffer cells and splenic stromal M phi, but was low or undetectable on blood monocytes and thymic, peritoneal, pleural, and bronchoalveolar M phi. SER therefore appeared to be expressed on certain M phi populations embedded in solid tissues but was largely absent from M phi recoverable by lavage. Its absence from monocytes implies that SER is acquired by M phi after entering tissues where it may perform adhesive functions. In bone marrow, SER on RBMM could interact with an appropriate sialylated ligand on murine hematopoietic cells, and could influence their rate of growth and differentiation.


Transfusion requirements in septic shock (TRISS) trial - comparing the effects and safety of liberal versus restrictive red blood cell transfusion in septic shock patients in the ICU: protocol for a randomised controlled trial.
Transfusion of red blood cells (RBC) is recommended in septic shock and the majority of these patients receive RBC transfusion in the intensive care unit (ICU). However, benefit and harm of RBCs have not been established in this group of high-risk patients. The Transfusion Requirements in Septic Shock (TRISS) trial is a multicenter trial with assessor-blinded outcome assessment, randomising 1,000 patients with septic shock in 30 Scandinavian ICUs to receive transfusion with pre-storage leuko-depleted RBC suspended in saline-adenine-glucose and mannitol (SAGM) at haemoglobin level (Hb) of 7 g/dl or 9 g/dl, stratified by the presence of haematological malignancy and centre. The primary outcome measure is 90-day mortality. Secondary outcome measures are organ failure, ischaemic events, severe adverse reactions (SARs: anaphylactic reaction, acute haemolytic reaction and transfusion-related circulatory overload, and acute lung injury) and mortality at 28 days, 6 months and 1 year.The sample size will enable us to detect a 9% absolute difference in 90-day mortality assuming a 45% event rate with a type 1 error rate of 5% and power of 80%. An interim analysis will be performed after 500 patients, and the Data Monitoring and Safety Committee will recommend the trial be stopped if a group difference in 90-day mortality with P ≤0.001 is present at this point. The TRISS trial may bridge the gap between clinical practice and the lack of efficacy and safety data on RBC transfusion in septic shock patients. The effect of restrictive versus liberal RBC transfusion strategy on mortality, organ failure, ischaemic events and SARs will be evaluated.


Virtual reality and consciousness inference in dreaming.
This article explores the notion that the brain is genetically endowed with an innate virtual reality generator that - through experience-dependent plasticity - becomes a generative or predictive model of the world. This model, which is most clearly revealed in rapid eye movement (REM) sleep dreaming, may provide the theater for conscious experience. Functional neuroimaging evidence for brain activations that are time-locked to rapid eye movements (REMs) endorses the view that waking consciousness emerges from REM sleep - and dreaming lays the foundations for waking perception. In this view, the brain is equipped with a virtual model of the world that generates predictions of its sensations. This model is continually updated and entrained by sensory prediction errors in wakefulness to ensure veridical perception, but not in dreaming. In contrast, dreaming plays an essential role in maintaining and enhancing the capacity to model the world by minimizing model complexity and thereby maximizing both statistical and thermodynamic efficiency. This perspective suggests that consciousness corresponds to the embodied process of inference, realized through the generation of virtual realities (in both sleep and wakefulness). In short, our premise or hypothesis is that the waking brain engages with the world to predict the causes of sensations, while in sleep the brain's generative model is actively refined so that it generates more efficient predictions during waking. We review the evidence in support of this hypothesis - evidence that grounds consciousness in biophysical computations whose neuronal and neurochemical infrastructure has been disclosed by sleep research.


A single methyltransferase YefA (RlmCD) catalyses both m5U747 and m5U1939 modifications in Bacillus subtilis 23S rRNA.
Methyltransferases that use S-adenosylmethionine (AdoMet) as a cofactor to catalyse 5-methyl uridine (m(5)U) formation in tRNAs and rRNAs are widespread in Bacteria and Eukaryota, and are also found in certain Archaea. These enzymes belong to the COG2265 cluster, and the Gram-negative bacterium Escherichia coli possesses three paralogues. These comprise the methyltransferases TrmA that targets U54 in tRNAs, RlmC that modifies U747 in 23S rRNA and RlmD that is specific for U1939 in 23S rRNA. The tRNAs and rRNAs of the Gram-positive bacterium Bacillus subtilis have the same three m(5)U modifications. However, as previously shown, the m(5)U54 modification in B. subtilis tRNAs is catalysed in a fundamentally different manner by the folate-dependent enzyme TrmFO, which is unrelated to the E. coli TrmA. Here, we show that methylation of U747 and U1939 in B. subtilis rRNA is catalysed by a single enzyme, YefA that is a COG2265 member. A recombinant version of YefA functions in an E. coli m(5)U-null mutant adding the same two rRNA methylations. The findings suggest that during evolution, COG2265 enzymes have undergone a series of changes in target specificity and that YefA is closer to an archetypical m(5)U methyltransferase. To reflect its dual specificity, YefA is renamed RlmCD.


A lysosome independent role for TFEB in activating DNA repair and inhibiting apoptosis in breast cancer cells.
Transcription factor EB (TFEB) is a master regulator of lysosomal biogenesis and autophagy with critical roles in several cancers. Lysosomal autophagy promotes cancer survival through the degradation of toxic molecules and the maintenance of adequate nutrient supply. Doxorubicin (DOX) is the standard of care treatment for triple-negative breast cancer (TNBC); however, chemoresistance at lower doses and toxicity at higher doses limit its usefulness. By targeting pathways of survival, DOX can become an effective antitumor agent. In this study, we examined the role of TFEB in TNBC and its relationship with autophagy and DNA damage induced by DOX. In TNBC cells, TFEB was hypo-phosphorylated and localized to the nucleus upon DOX treatment. TFEB knockdown decreased the viability of TNBC cells while increasing caspase-3 dependent apoptosis. Additionally, inhibition of the TFEB-phosphatase calcineurin sensitized cells to DOX-induced apoptosis in a TFEB dependent fashion. Regulation of apoptosis by TFEB was not a consequence of altered lysosomal function, as TFEB continued to protect against apoptosis in the presence of lysosomal inhibitors. RNA-Seq analysis of MDA-MB-231 cells with TFEB silencing identified a down-regulation in cell cycle and homologous recombination genes while interferon-γ and death receptor signaling genes were up-regulated. In consequence, TFEB knockdown disrupted DNA repair following DOX, as evidenced by persistent γH2A.X detection. Together, these findings describe in TNBC a novel lysosomal independent function for TFEB in responding to DNA damage.


Sequential glycan profiling at single cell level with the microfluidic lab-in-a-trench platform: a new era in experimental cell biology.
It is now widely recognised that the earliest changes that occur on a cell when it is stressed or becoming diseased are alterations in its surface glycosylation. Current state-of-the-art technologies in glycoanalysis include mass spectrometry, protein microarray formats, techniques in cytometry and more recently, glyco-quantitative polymerase chain reaction (Glyco-qPCR). Techniques for the glycoprofiling of the surfaces of single cells are either limited to the analysis of large cell populations or are unable to handle multiple and/or sequential probing. Here, we report a novel approach of single live cell glycoprofiling enabled by the microfluidic "Lab-in-a-Trench" (LiaT) platform for performing capture and retention of cells, along with shear-free reagent loading and washing. The significant technical improvement on state-of-the-art is the demonstration of consecutive, spatio-temporally profiling of glycans on a single cell by sequential elution of the previous lectin probe using their corresponding free sugar. We have qualitatively analysed glycan density on the surface of individual cells. This has allowed us to qualitatively co-localise the observed glycans. This approach enables exhaustive glycoprofiling and glycan mapping on the surface of individual live cells with multiple lectins. The possibility of sequentially profiling glycans on cells will be a powerful new tool to add to current glycoanalytical techniques. The LiaT platform will enable cell biologists to perform many high sensitivity assays and also will also make a significant impact on biomarker research.


Mechanisms for segregating T cell receptor and adhesion molecules during immunological synapse formation in Jurkat T cells.
T cells interacting with antigen-presenting cells (APCs) form an "immunological synapse" (IS), a bull's-eye pattern composed of a central supramolecular activation cluster enriched with T cell receptors (TCRs) surrounded by a ring of adhesion molecules (a peripheral supramolecular activation cluster). The mechanism responsible for segregating TCR and adhesion molecules remains poorly understood. Here, we show that immortalized Jurkat T cells interacting with a planar lipid bilayer (mimicking an APC) will form an IS, thereby providing an accessible model system for studying the cell biological processes underlying IS formation. We found that an actin-dependent process caused TCR and adhesion proteins to cluster at the cell periphery, but these molecules appeared to segregate from one another at the earliest stages of microdomain formation. The TCR and adhesion microdomains attached to actin and were carried centripetally by retrograde flow. However, only the TCR microdomains penetrated into the actin-depleted cell center, whereas the adhesion microdomains appeared to be unstable without an underlying actin cytoskeleton. Our results reveal that TCR and adhesion molecules spatially partition from one another well before the formation of a mature IS and that differential actin interactions help to shape and maintain the final bull's-eye pattern of the IS.