Luteolin triggers global changes in the microglial transcriptome leading to a unique anti-inflammatory and neuroprotective phenotype.
Luteolin, a plant derived flavonoid, exerts a variety of pharmacological activities and anti-oxidant properties associated with its capacity to scavenge oxygen and nitrogen species. Luteolin also shows potent anti-inflammatory activities by inhibiting nuclear factor kappa B (NFkB) signaling in immune cells. To better understand the immuno-modulatory effects of this important flavonoid, we performed a genome-wide expression analysis in pro-inflammatory challenged microglia treated with luteolin and conducted a phenotypic and functional characterization. Resting and LPS-activated BV-2 microglia were treated with luteolin in various concentrations and mRNA levels of pro-inflammatory markers were determined. DNA microarray experiments and bioinformatic data mining were performed to capture global transcriptomic changes following luteolin stimulation of microglia. Extensive qRT-PCR analyses were carried out for an independent confirmation of newly identified luteolin-regulated transcripts. The activation state of luteolin-treated microglia was assessed by morphological characterization. Microglia-mediated neurotoxicity was assessed by quantifying secreted nitric oxide levels and apoptosis of 661W photoreceptors cultured in microglia-conditioned medium. Luteolin dose-dependently suppressed pro-inflammatory marker expression in LPS-activated microglia and triggered global changes in the microglial transcriptome with more than 50 differentially expressed transcripts. Pro-inflammatory and pro-apoptotic gene expression was effectively blocked by luteolin. In contrast, mRNA levels of genes related to anti-oxidant metabolism, phagocytic uptake, ramification, and chemotaxis were significantly induced. Luteolin treatment had a major effect on microglial morphology leading to ramification of formerly amoeboid cells associated with the formation of long filopodia. When co-incubated with luteolin, LPS-activated microglia showed strongly reduced NO secretion and significantly decreased neurotoxicity on 661W photoreceptor cultures. Our findings confirm the inhibitory effects of luteolin on pro-inflammatory cytokine expression in microglia. Moreover, our transcriptomic data suggest that this flavonoid is a potent modulator of microglial activation and affects several signaling pathways leading to a unique phenotype with anti-inflammatory, anti-oxidative, and neuroprotective characteristics. With the identification of several novel luteolin-regulated genes, our findings provide a molecular basis to understand the versatile effects of luteolin on microglial homeostasis. The data also suggest that luteolin could be a promising candidate to develop immuno-modulatory and neuroprotective therapies for the treatment of neurodegenerative disorders.


Contact lens articles in the literature: an update.
To determine if the pattern of publication of contact lens articles in the ophthalmic literature has changed in recent years. We searched the literature of the past 5 years and reviewed the abstracts of all published papers that presented information on contact lenses. The journals reviewed included the American Journal of Ophthalmology, Archives of Ophthalmology, Ophthalmology, and The CLAO Journal. We determined whether articles were positive, negative, or neutral. We reviewed a total of 1,189,451 articles and 48,811 (4.10%) eye articles; of the latter, 987 (2.02%) discussed contact lenses. Only 86 total contact lens articles were published in the three major ophthalmology journals, of which almost 40% presented negative information (e.g., complications of contact lens usage). The CLAO Journal published 136 contact lens articles during the same time period (39.7% were positive and 30.1% were negative). In the past 5 years, fewer contact lens articles have appeared in the ophthalmic literature than had appeared in the previous five years, and, in addition, more of these articles were negative. The CLAO Journal continues to publish the largest number of contact lens articles, although the percentage of positive articles has decreased somewhat, from 50% to 39.7%.


Diagnosis of Pulmonary Hypertension with Cardiac MRI: Derivation and Validation of Regression Models.
Purpose To derive and test multiparametric cardiac MRI models for the diagnosis of pulmonary hypertension (PH). Materials and Methods Images and patient data from consecutive patients suspected of having PH who underwent cardiac MRI and right-sided heart catheterization (RHC) between 2012 and 2016 were retrospectively reviewed. Of 2437 MR images identified, 603 fit the inclusion criteria. The mean patient age was 61 years (range, 18-88 years; mean age of women, 60 years [range, 18-84 years]; mean age of men, 62 years [range, 22-88 years]). In the first 300 patients (derivation cohort), cardiac MRI metrics that showed correlation with mean pulmonary arterial pressure (mPAP) were used to create a regression algorithm. The performance of the model was assessed in the 303-patient validation cohort by using receiver operating characteristic (ROC) and χ2 analysis. Results In the derivation cohort, cardiac MRI mPAP model 1 (right ventricle and black blood) was defined as follows: -179 + loge interventricular septal angle × 42.7 + log10 ventricular mass index (right ventricular mass/left ventricular mass) × 7.57 + black blood slow flow score × 3.39. In the validation cohort, cardiac MRI mPAP model 1 had strong agreement with RHC-measured mPAP, an intraclass coefficient of 0.78, and high diagnostic accuracy (area under the ROC curve = 0.95; 95% confidence interval [CI]: 0.93, 0.98). The threshold of at least 25 mm Hg had a sensitivity of 93% (95% CI: 89%, 96%), specificity of 79% (95% CI: 65%, 89%), positive predictive value of 96% (95% CI: 93%, 98%), and negative predictive value of 67% (95% CI: 53%, 78%) in the validation cohort. A second model, cardiac MRI mPAP model 2 (right ventricle pulmonary artery), which excludes the black blood flow score, had equivalent diagnostic accuracy (ROC difference: P = .24). Conclusion Multiparametric cardiac MRI models have high diagnostic accuracy in patients suspected of having pulmonary hypertension. Published under a CC BY 4.0 license. Online supplemental material is available for this article. See also the editorial by Colletti in this issue.


Injection of the subacromial bursa in patients with rotator cuff syndrome: a prospective, randomized study comparing the effectiveness of different routes.
Rotator cuff syndrome is often treated with subacromial injection of corticosteroid and local anesthetic. It has not been established if the common injection routes of the bursa are equally accurate. We conducted a prospective clinical trial involving seventy-five shoulders in seventy-five patients who were randomly assigned to receive a subacromial injection through an anterior, lateral, or posterior route with respect to the acromion. An experienced physician performed the injections, which contained radiopaque contrast medium, corticosteroid, and local anesthetic. After the injection, a musculoskeletal radiologist, blinded to the injection route, interpreted all of the radiographs. The rate of accuracy varied with the route of injection, with a rate of 56% for the posterior route, 84% for the anterior route, and 92% for the lateral route (p = 0.006; chi-square test). The accuracy of injection through the posterior route was significantly lower than that through either the anterior or the lateral route (p < 0.05 for both comparisons; Poisson regression). In addition, the accuracy of injection was significantly lower in females than in males (p < 0.006; chi-square test). Among males, no differences between the routes were noted (with accuracy rates of 89% for the posterior route, 92% for the anterior route, and 93% for the lateral route). Among females, however, the accuracy of injection was lower for the posterior route than for either the anterior or the lateral route (with accuracy rates of 38% for the posterior route, 77% for the anterior route, and 91% for the lateral route) (p < 0.05). The anterior and lateral routes of subacromial bursal injection were more accurate than the posterior route. The accuracy of subacromial bursal injection was significantly different between males and females, mainly because of a lower accuracy of bursal injection with use of the posterior route in females. The present study suggests that the posterior route is the least accurate method for injection of the subacromial bursa in females.


Entropic depletion in colloidal suspensions and polymer liquids: role of nanoparticle surface topography.
We employ a hybrid Monte Carlo plus integral equation theory approach to study how dense fluids of small nanoparticles or polymer chains mediate entropic depletion interactions between topographically rough particles where all interaction potentials are hard core repulsion. The corrugated particle surfaces are composed of densely packed beads which present variable degrees of controlled topographic roughness and free volume associated with their geometric crevices. This pure entropy problem is characterized by competing ideal translational and (favorable and unfavorable) excess entropic contributions. Surface roughness generically reduces particle depletion aggregation relative to the smooth hard sphere case. However, the competition between ideal and excess packing entropy effects in the bulk, near the particle surface and in the crevices, results in a non-monotonic variation of the particle-monomer packing correlation function as a function of the two dimensionless length scale ratios that quantify the effective surface roughness. As a result, the inter-particle potential of mean force (PMF), second virial coefficient, and spinodal miscibility volume fraction vary non-monotonically with the surface bead to monomer diameter and particle core to surface bead diameter ratios. A miscibility window is predicted corresponding to an optimum degree of surface roughness that completely destroys depletion attraction resulting in a repulsive PMF. Variation of the (dense) matrix packing fraction can enhance or suppress particle miscibility depending upon the amount of surface roughness. Connecting the monomers into polymer chains destabilizes the system via enhanced contact depletion attraction, but the non-monotonic variations with surface roughness metrics persist.


A comparison of patients requiring orthognathic treatment who subsequently refused or accepted surgery.
Forty-four patients who required combined orthodontic-surgical procedures were psychologically assessed in relation to stress, self-esteem, introspectiveness, and body satisfaction. Thirty patients accepted the surgical procedures, and 14 refused surgery. These two groups of patients were compared with respect to their psychologic profiles; statistical analysis detected no significant differences.


Cardio-respiratory modeling in fishes and the consequences of the evolution of airbreathing.
The microcirculation of the respiratory organ of water and air breathing vertebrates is similar and can be described as sheet flow. The gross morphologies of the systems, however, are very different and reflect the properties of the medium. The fish heart has a single ventricle that forces blood first through the gills and then through the body. The pressure in the gills is higher than in the systemic circulation, the reverse of the situation seen in mammals. The gill epithelium is thicker than that in the lung and is involved in ionic and acid-base functions carried out in the kidney of mammals. Gills stick together in air. Therefore, fish breathe air using some other structure, such as the gut or mouth, the swimbladder, or the skin. The gills are retained for carbon dioxide excretion and ion and acid-base regulation. This results in a separation of oxygen uptake and carbon dioxide excretion. The gills are often modified in air-breathing fish such that venous blood flows to well developed gills for carbon dioxide and acid excretion, whereas oxygenated blood flow bypasses the gills. This is the beginning of a separation of flows in the heart which is more highly developed in amphibians and reptiles and complete in mammals. The loss of gills requires transfer of ionic and acid base regulation processes to the skin in amphibia and to the kidney in reptiles and mammals, allowing a completely terrestrial existence. The organization of the venous system is influenced by the degree of support offered by the medium.(ABSTRACT TRUNCATED AT 250 WORDS)


Neural Networks Push the Limits of Luminescence Lifetime Nanosensing.
Luminescence lifetime-based sensing is ideally suited to monitor biological systems due to its minimal invasiveness and remote working principle. Yet, its applicability is limited in conditions of low signal-to-noise ratio (SNR) induced by, e.g., short exposure times and presence of opaque tissues. Herein this limitation is overcome by applying a U-shaped convolutional neural network (U-NET) to improve luminescence lifetime estimation under conditions of extremely low SNR. Specifically, the prowess of the U-NET is showcased in the context of luminescence lifetime thermometry, achieving more precise thermal readouts using Ag2 S nanothermometers. Compared to traditional analysis methods of decay curve fitting and integration, the U-NET can extract average lifetimes more precisely and consistently regardless of the SNR value. The improvement achieved in the sensing performance using the U-NET is demonstrated with two experiments characterized by extreme measurement conditions: thermal monitoring of free-falling droplets, and monitoring of thermal transients in suspended droplets through an opaque medium. These results broaden the applicability of luminescence lifetime-based sensing in fields including in vivo experimentation and microfluidics, while, hopefully, spurring further research on the implementation of machine learning (ML) in luminescence sensing.


'But I'm not like that': young men's navigation of normative masculinities in a marginalised urban community in Paraguay.
Young men often define themselves and their masculine identity through romantic and sexual relationships, and their resulting sexual decisions can affect their successful transition into adulthood, as well as STI, HIV and pregnancy rates. This paper looks at how young Paraguayan men's peer groups, family and masculine identity formation influence their behaviours in sexual and romantic relationships. In Asunción, Paraguay, we conducted five focus-group discussions (FGDs) examining neighbourhood norms in 2010, with male peer groups ranging in age from 14 to 19 years. We then interviewed half the members from each peer group to examine their relationships with friends, family and young women and their beliefs about existing gender norms. Young men described two types of masculine norms, 'partner/provider' and macho, and two types of romantic relationships, 'casual' and 'formal'. The language used to describe each spectrum of behaviours was often concordant and highlights the connection between masculine norms and romantic relationships. The perceived norms for the neighbourhood were more macho than the young men's reported behaviours. Norms cannot change unless young men speak out about their non-normative behaviours. This provides evidence for more research on the formation, meaning and transformation of male gender norms.


Phylogeography of skinks (Chalcides) in the Canary Islands inferred from mitochondrial DNA sequences.
Mitochondrial DNA (mtDNA) evolution was investigated in skinks of the genus Chalcides found in the Canary Islands (Ch. sexlineatus, Ch. viridanus and Ch. simonyi), together with some North African congenerics (Ch. polylepis and Ch. mionecton). Several sites were included within islands to cover areas of known within-island geographical variation in morphology. Skinks from the islands of El Hierro and La Gomera appear to be sister taxa. The relationships between this clade and the Tenerife and Gran Canarian skinks were not fully resolved, although the best working hypothesis indicated monophyly with the former, with the latter forming a closely related outgroup. Ch. simonyi from Fuerteventura was more distantly related to the Western Canary Island skinks and did not show close relationships with the North African species Ch. mionecton and Ch. polylepis. Possible colonization sequences for the four most Western Canary Islands were considered. El Hierro appears to have been colonized relatively recently from La Gomera, commensurate with the recent origin of this island, while dispersal between La Gomera and Tenerife and between Gran Canaria and Tenerife or La Gomera appears to have taken place considerably earlier. Substantial within-island haplotype divergence was found in Gran Canaria and Tenerife. This may be a result of recent periods of intense volcanic activity found within these two islands. Lower levels of within-island differentiation are found in La Gomera and El Hierro and may be explained by lower levels of volcanic activity during recent geological history and a more recent colonization, respectively.


ADHD: Insurance and Mental Health Service Use.
We describe mental health service use by insurance among children aged 4 to 17 with diagnosed attention-deficit/ hyperactivity disorder (ADHD). Using parent reports from 2010-2013 National Health Interview Survey, we estimate the percentage that received services for emotional and behavioral difficulties (EBD): medication, other nonmedication services, and none (neither medication nor other nonmedication services). Among children with diagnosed ADHD, 56.0% had used medication for EBD, 39.8% had contact with a mental health professional, 32.2% had contact with a general doctor about the child's EBD, and 20.4% received special education services for EBD. Medication use was more often reported for privately or publicly insured children than uninsured children ( P < .001), and uninsured children more often received no services ( P < .001). Publicly insured children were more likely than privately insured children to receive other nonmedication services ( P < .001). Less than a third (28.9%) of all children received no services as compared to almost half (48.8%) of uninsured children.


Global and local regulation of clathrin-coated pit dynamics detected on patterned substrates.
Live-cell imaging of individual clathrin-coated pit (CCP) dynamics has revealed a broad variation in their internalization kinetics, but the functional significance and mechanistic underpinnings of this heterogeneity remain unknown. One contributing factor may be the spatial variations in the underlying actin cortex. To test this, we cultured cells on fibronectin (Fn) micropatterned substrates to vary the cortical actin mechanics in a defined manner. Under these conditions, stress fibers became organized to bridge adhesive islands, creating spatial heterogeneity in the cortical actin architecture. CCP lifetimes within the Fn-coated islands were selectively prolonged. This differential effect was not due to adherence to Fn-coated surfaces, and was not observed in cells grown on patterned surfaces that did not induce organized stress fiber assembly. Pharmacological agents that lower cortical tension selectively lowered CCP lifetimes within Fn islands, thus abolishing the spatial heterogeneity in the CCP dynamics. Although we cannot rule out the possibility that other factors might locally affect CCP dynamics at Fn islands, our data suggest that localized modulation in cortical tension may spatially regulate clathrin-mediated endocytosis.


Risk of persistent growth impairment after alternate-day prednisone treatment in children with cystic fibrosis.
It is uncertain whether the growth impairment that occurs in children during long-term treatment with glucocorticoids persists after the medication is discontinued and ultimately affects adult height. We evaluated growth six to seven years after alternate-day treatment with prednisone had been discontinued in 224 children 6 to 14 years of age with cystic fibrosis who had participated in a multicenter trial of this therapy from 1986 through 1991. Of the children, 151 had been randomly assigned to receive prednisone (either 1 or 2 mg per kilogram of body weight) and 73 to receive placebo. We obtained data on growth up to 1997 from the Cystic Fibrosis Foundation Patient Registry and standardized the data to sex- and age-specific norms from the National Center for Health Statistics. We used z scores to compare growth patterns among treatment groups. In 1997, 68 percent of the patients were 18 years of age or older. The z scores for height declined during prednisone therapy; catch-up growth began two years after treatment with prednisone was discontinued. Among the boys, the z scores for height in those treated with prednisone remained lower than the scores for those who received placebo (P=0.02). The mean heights for boys 18 years of age or older were 4 cm less in the prednisone groups than in the placebo group, an equivalent of 13 percentile points (P=0.03). Among the girls, differences in height between those who were treated with prednisone and those who received placebo were no longer present two to three years after prednisone therapy was discontinued. Among children with cystic fibrosis who have received alternate-day treatment with prednisone, boys, but not girls, have persistent growth impairment after treatment is discontinued.


Behavioral studies relevant to vaccine trial preparation: an introduction.
Preparations for large-scale trials to test the efficacy of candidate HIV vaccines can benefit in several crucial ways from a targeted program of behavioral and social research. Randomized field experiments testing alternative procedures for the recruitment and retention of subjects can help identify research procedures that will ensure adequate sample sizes while minimizing sample attrition over time. Similarly, assuring that subjects accurately comprehend the potential risks of participation will require more than simply presenting scientifically accurate information. Ensuring both the adequacy and appropriateness of risk communications as well as the accuracy of subject perception of risks (across the social and cultural milieux in which vaccine trials will be undertaken) is a critical task. Ethnographic and behavioral studies can help to ensure that our obligation to obtain truly informed consent from our research subjects is fully met and documented. Monitoring risk behaviors over the course of the vaccine trials could also benefit from strategic investments in new technologies developed by social researchers to permit the collection of sensitive personal data while affording complete privacy to subjects. These new measurement technologies include procedures that permit private data collection (without a human interviewer) in any spoken language and without requiring that subjects be literate.


Different presynaptic roles of synapsins at excitatory and inhibitory synapses.
The functions of synapsins were examined by characterizing the phenotype of mice in which all three synapsin genes were knocked out. Although these triple knock-out mice were viable and had normal brain anatomy, they exhibited a number of behavioral defects. Synaptic transmission was altered in cultured neurons from the hippocampus of knock-out mice. At excitatory synapses, loss of synapsins did not affect basal transmission evoked by single stimuli but caused a threefold increase in the rate of synaptic depression during trains of stimuli. This suggests that synapsins regulate the reserve pool of synaptic vesicles. This possibility was examined further by measuring synaptic vesicle density in living neurons transfected with green fluorescent protein-tagged synaptobrevin 2, a marker of synaptic vesicles. The relative amount of fluorescent synaptobrevin was substantially lower at synapses of knock-out neurons than of wild-type neurons. Electron microscopy also revealed a parallel reduction in the number of vesicles in the reserve pool of vesicles >150 nm away from the active zone at excitatory synapses. Thus, synapsins are required for maintaining vesicles in the reserve pool at excitatory synapses. In contrast, basal transmission at inhibitory synapses was reduced by loss of synapsins, but the kinetics of synaptic depression were unaffected. In these terminals, there was a mild reduction in the total number of synaptic vesicles, but this was not restricted to the reserve pool of vesicles. Thus, synapsins maintain the reserve pool of glutamatergic vesicles but regulate the size of the readily releasable pool of GABAergic vesicles.


Deep characterization of paired chromatin and transcriptomes in four immune cell types from multiple sclerosis patients.
Background: The putative involvement of chromatin states in multiple sclerosis (MS) is thus far unclear. Here we determined the association of chromatin-accessibility with concurrent genetic, epigenetic and transcriptional events. Material & methods: We generated paired assay for transposase-accessible chromatin sequencing and RNA-sequencing profiles from sorted blood immune CD4+ and CD8+ T cells, CD14+ monocytes and CD19+ B cells from healthy controls (HCs) and MS patients. Results: We identified differentially accessible regions between MS patients and HCs, primarily in CD4+ and CD19+. CD4+ regions were enriched for MS-associated single nucleotide polymorphisms and differentially methylated loci. In the vicinity of differentially accessible regions of CD4+ cells, 42 differentially expressed genes were identified. The top two dysregulated genes identified in this multilayer analysis were CCDC114 and SERTAD1. Conclusion: These findings provide new insight into the primary role of CD4+ and CD19+ cells in MS.


Left ventricular hypertrophy in athletes: How to differentiate between hypertensive heart disease and athlete's heart.
Athlete's heart is typically accompanied by a remodelling of the cardiac chambers induced by exercise. However, although competitive athletes are commonly considered healthy, they can be affected by cardiac disorders characterised by an increase in left ventricular mass and wall thickness, such as hypertension. Unfortunately, training-induced increase in left ventricular mass, wall thickness, and atrial and ventricular dilatation observed in competitive athletes may mimic the pathological remodelling of pathological hypertrophy. As a consequence, distinguishing between athlete's heart and hypertension can sometimes be challenging. The present review aimed to focus on the differential diagnosis between hypertensive heart disease and athlete's heart, providing clinical information useful to distinguish between physiological and pathological remodelling.


Discovering potassium channel blockers from synthetic compound database by using structure-based virtual screening in conjunction with electrophysiological assay.
Potassium ion (K+) channels are attractive targets for drug discovery because of the essential roles played in biological systems. However, high-throughput screening (HTS) cannot be used to screen K+ channel blockers. To overcome this disadvantage of HTS, we have developed a virtual screening approach for discovering novel blockers of K+ channels. On the basis of a three-dimensional model of the eukaryotic K+ channels, molecular docking-based virtual screening was employed to search the chemical database MDL Available Chemicals Directory (ACD). Compounds were ranked according to their relative binding energy, favorable shape complementarity, and potential to form hydrogen bonds with the outer mouth of the K+ channel model. Twenty candidate compounds selected from the virtual screening were examined using the whole-cell voltage-clamp recording in rat dissociated hippocampal neurons. Among them, six compounds (5, 6, 8, 18-20) potently blocked both the delayed rectifier (IK) and fast transient K+ currents (IA). When applied externally, these six compounds preferentially blocked IK with potencies 2- to 500-fold higher than that of tetraethylammonium chloride. Intracellular application of the six compounds had no effect on both K+ currents. In addition, the interaction models and binding free energy calculations demonstrated that hydrophobic interaction and solvent effects play important roles in the inhibitory activities of these compounds. The results demonstrated that structure-based computer screening strategy could be used to identify novel, structurally diverse compounds targeting the pore binding pocket of the outer mouth of voltage-gated K+ channels. This study provides an alternative way of finding new blockers of voltage-gated K+ channels, while the techniques for high-throughput screening of K+ channel drugs remain in development.


Gene expression profiling identifies molecular subgroups among nodal peripheral T-cell lymphomas.
The classification of peripheral T-cell lymphomas (PTCL) is still a matter of debate. To establish a molecular classification of PTCL, we analysed 59 primary nodal T-cell lymphomas using cDNA microarrays, including 56 PTCL and three T-lymphoblastic lymphoma (T-LBL). The expression profiles could discriminate angioimmunoblastic lymphoma, anaplastic large-cell lymphoma and T-LBL. In contrast, cases belonging to the broad category of 'PTCL, unspecified' (PTCL-U) did not share a single molecular profile. Using a multiclass predictor, we could separate PTCL-U into three molecular subgroups called U1, U2 and U3. The U1 gene expression signature included genes known to be associated with poor outcome in other tumors, such as CCND2. The U2 subgroup was associated with overexpression of genes involved in T-cell activation and apoptosis, including NFKB1 and BCL-2. The U3 subgroup was mainly defined by overexpression of genes involved in the IFN/JAK/STAT pathway. It comprised a majority of histiocyte-rich PTCL samples. Gene Ontology annotations revealed different functional profile for each subgroup. These results suggest the existence of distinct subtypes of PTCL-U with specific molecular profiles, and thus provide a basis to improve their classification and to develop new therapeutic targets.


Utility of genetic testing for therapeutic decision-making in adults with epilepsy.
Genetic testing has become a routine part of the diagnostic workup in children with early onset epilepsies. In the present study, we sought to investigate a cohort of adult patients with epilepsy, to determinate the diagnostic yield and explore the gain of personalized treatment approaches in adult patients. Two hundred patients (age span = 18-80 years) referred for diagnostic gene panel testing at the Danish Epilepsy Center were included. The vast majority (91%) suffered from comorbid intellectual disability. The medical records of genetically diagnosed patients were mined for data on epilepsy syndrome, cognition, treatment changes, and seizure outcome following the genetic diagnosis. We found a genetic diagnosis in 46 of 200 (23%) patients. SCN1A, KCNT1, and STXBP1 accounted for the greatest number of positive findings (48%). More rare genetic findings included SLC2A1, ATP6A1V, HNRNPU, MEF2C, and IRF2BPL. Gene-specific treatment changes were initiated in 11 of 46 (17%) patients (one with SLC2A1, 10 with SCN1A) following the genetic diagnosis. Ten patients improved, with seizure reduction and/or increased alertness and general well-being. With this study, we show that routine diagnostic testing is highly relevant in adults with epilepsy. The diagnostic yield is similar to previously reported pediatric cohorts, and the genetic findings can be useful for therapeutic decision-making, which may lead to better seizure control, ultimately improving quality of life.


Pretreatment of diabetic rats with lipoic acid improves healing of subsequently-induced abrasion wounds.
The etiology of delayed or impaired wound-healing in diabetic individuals is multifactoral, but peripheral vascular dysfunction is an underlying factor in the majority of cases. Recent studies have shown that lipoic acid improves vascular function in diabetic skin and reduces the symptoms associated with the diabetic peripheral neuropathy. In this study, rats were made diabetic with streptozotocin (STZ) and treated systemically on alternative days with lipoic acid (100 mg/kg given via intraperitoneal injection) for 8 weeks. Untreated STZ-diabetic rats and non-diabetic rats served as control. At the end of the 8-week period, rats from all the three groups were subjected to abrasion wound formation. Skin wounds healed more rapidly in untreated non-diabetic rats than in the untreated diabetic rats. Wounds in lipoic acid-treated diabetic rats healed more rapidly than wounds in untreated diabetic rats. Subsequent in vitro studies demonstrated that lipoic acid protected endothelial cells from oxidant injury. At the same time, lipoic acid had no apparent effect on endothelial cell proliferation and had no measurable effect on fibroblast function (proliferation, collagen synthesis and matrix metalloproteinase expression). These findings suggest that prophylactic use of lipoic acid might be useful in preventing the development of non-healing skin ulcers from minor traumas in at-risk skin.


Phenotypic and genotypic characterization of Vibrio vulnificus: proposal for the substitution of the subspecific taxon biotype for serovar.
The classification of Vibrio vulnificus strains into two biotypes has been maintained on the basis of phenotypic properties and eel virulence. Biotype 2 is virulent for eels, negative for the indole reaction, and serologically homogeneous (serogroup E), whereas strains of biotype 1 are avirulent, indole positive, and serologically heterogeneous. In the present study, we phenotypically and genotypically characterized 21 V. vulnificus isolates, recovered mainly from northern Europe, by comparing them with reference strains of both biotypes to look for new isolates of biotype 2. The results of this work revealed that the majority of isolates virulent for eels presented phenotypic traits previously considered characteristics of biotype 2 and specific ribotypes with HindIII. However, among the new isolates we found (i) a serogroup E strain virulent for eels but indole positive and (ii) one isolate not belonging to serogroup E but pathogenic for eels. Since no biochemical test for specific serogroup can with certainty be associated with eel virulence, we propose to classify V. vulnificus strains into serovars instead of biotypes. Thus, we suggest serovar E as the denomination of those strains previously classified as biotype 2. Finally, the occurrence of serogroup E in eels cultured in Norway and Sweden, as well as from human infections and shrimp, has been demonstrated.


Small dense LDL-cholesterol relative to LDL-cholesterol is a strong independent determinant of hypoadiponectinemia in metabolic syndrome.
Small dense low-density lipoprotein (sd-LDL) is an atherogenic lipoprotein closely associated with an increased risk of cardiovascular diseases. However, a precise analysis of the actual amount of sd-LDL-cholesterol (sd-LDL-C) in patients with metabolic syndrome (MS) has not been performed. Among 214 patients enrolled in the present study, 101 patients (47%) met the Japanese MS criteria. The serum levels of sd-LDL-C determined with a dual detection HPLC system were higher in MS than non-MS patients, while total cholesterol and low-density lipoprotein-cholesterol (LDL-C) were similar between MS and non-MS patients. Compared with the sd-LDL-C and LDL-C level, the ratio sd-LDL-C/LDL-C was more closely correlated with various parameters associated with MS. A multivariate regression analysis revealed that the ratio sd-LDL-C/LDL-C is the strongest independent determinant of hypoadiponectinemia. Furthermore, weight reduction therapy through diet and exercise rapidly decreased LDL-C but slowly decreased sd-LDL-C. At 12 months after the therapy, weight reduction led to a significant decrease in the ratio sd-LDL-C/LDL-C in tandem with increasing adiponectin levels. These findings demonstrate that the ratio sd-LDL-C/LDL-C is tightly connected with hypoadiponectinemia and provides a useful clinical indicator for MS. The results also suggest that the elevation of this ratio can be modulated by long-term lifestyle changes.


Bronchoscopic management of a primary endobronchial salivary epithelial-myoepithelial carcinoma: A case report.
Here, we discussed a 55 y/o African man who recently immigrated from Nigeria to the United States and who presented to Parkland Memorial Hospital with a productive, intermittent cough of one year duration. The cough was associated with shortness of breath and chest pain. Cough was not associated with voice hoarseness, hemoptysis, melanoptysis, and wheezing. He had a computed tomography (CT) scan of the chest that showed a 1.9 cm mass in the right main stem bronchus with ipsilateral right lower lobe consolidation and bronchiectasis. The patient was seen by pulmonology who recommended bronchoscopy for diagnosis and possible intervention. Bronchoscopy showed a 90% obstructing mass in the proximal right mainstem bronchus and bronchus intermedius. The mass was large and endobronchial, circumferential, exophytic, and polypoid. The decision was made to undergo bronchoscopic tumor ablation using electrocautery snare, argon plasma coagulation (APC), suction, and forceps. The tumor was successful ablated. Microscopic examination revealed eosinophilic ducts tightly coupled with a surrounding layer of clear cell myoepithelial cells and the diagnosis of epithelial-myoepithelial carcinoma (EMC) of the lung was made. The patient was discharged from the hospital with scheduled outpatient visits for monitoring of the carcinoma by pulmonology and thoracic surgery. Unfortunately, he was lost to follow up.


Endoscopically placed nasoenteral feeding tubes. Indications and techniques.
We reviewed indications, techniques, and results for endoscopic placement of nasoenteral feeding tubes by analyzing records of 29 hospitalized patients who had undergone 41 tube placements. Indications included gastric stasis after gastric surgery (11 patients), gastroparesis and reflux or both in critically ill trauma patients (10), stricture secondary to vertical banded gastroplasty (3), and partial gastric outlet obstruction (5). Two endoscopic techniques were used. The guidewire method in which a flexible, Silastic feeding tube is advanced over the guidewire to the desired site had 82 per cent success (22 of 27). Adjunct fluoroscopy achieved a 94 per cent success rate (17 of 18); success without fluoroscopy was 56 per cent (5 of 9). The pull-along method attached a suture to the feeding tube tip which was grasped with forceps and moved to the correct location (93% success; 13 of 14). Fluoroscopy was not used with this method. Feeding tubes were successfully placed in 35 of 41 attempts (85%) in 27 of 29 patients. Failures were caused by an inability to intubate the efferent limb of a gastrojejunostomy (2) and unrecognized guidewire movement (4). Tube use ranged from one day to six months. We conclude that endoscopic placement of nasoenteral feeding tubes is an effective means to establish a route for enteral nutrition in selected patients.


[Mechanism of reduced binding capacity of plasma corticosteroid-binding globulin in acute radiation sickness].
Injection of various doses of thyroxin to rats was shown to increase a binding capacity of corticosteroid-binding globulin (CSG) of blood plasma after irradiation with doses of 650 and 800 R. The effect of thyroxin was most pronounced at a mid-lethal dose of 650 R and at the early stage of radiation sickness.


Thioacetamide intoxication triggers transcriptional up-regulation but enzyme inactivation of UDP-glucuronosyltransferases.
Thioacetamide (TAA) is a potent hepatotoxicant and has been widely used to develop experimental liver fibrosis/cirrhosis models. Although the liver toxicity of TAA has been extensively studied, little is known about its potential influence on UDP-glucuronosyltransferases (UGTs) associated with the development of liver fibrosis. The study presented here aimed to uncover the regulation patterns of UGTs in TAA-induced liver fibrosis of rats. Potential counteracting effects of hepatoprotective agents were also determined. TAA treatment for 8 weeks induced a significant transcriptional up-regulation of the major UGT isoforms, including UGT1A1, UGT1A6, and UGT2B1, accompanied with the dramatic elevations of most typical serum biomarkers of liver function and fibrosis scores. Upon TAA intoxication, the mRNA and protein levels of the major UGT isoforms were increased to 1.5- to 2.5-fold and 2.5- to 3.3-fold of that of the normal control, respectively. The hepatoprotective agents Schisandra spp. lignans extract and dimethyl diphenyl bicarboxylate could largely abolish TAA-induced up-regulation of all three UGT isoforms. However, enzyme activities of UGTs remained unchanged after TAA treatment. The dissociation of protein expression and enzyme activity could possibly be attributed to the inactivating effects of TAA, upon a NADPH-dependent bioactivation, on UGTs. This study suggests that the transcriptional up-regulation of UGTs may be an alternative mechanism of their preserved activities in liver fibrosis/cirrhosis.


Transoral ultrasonographic evaluation of carotid flow in predicting cerebral hemodynamics after carotid endarterectomy.
We investigated whether measurement of blood flow in the extracranial distal internal carotid artery (ICA) by transoral carotid ultrasonography (TOCU) can predict the cerebral hemodynamics and the hemodynamic effect of carotid endarterectomy (CEA) in patients with unilateral carotid stenosis. Forty-nine patients with unilateral ICA stenosis who underwent CEA were studied. Preoperative blood flow in the poststenotic portion of the extracranial ICA was studied by using TOCU. Regional cerebral blood flow (rCBF) and vasoreactivity to acetazolamide (VR) in the territory of the middle cerebral artery were investigated by using single-photon emission CT (SPECT) before, 2 weeks after, and 3 months after CEA. Doppler flow velocities in the extracranial distal ICA measured transorally by TOCU correlated with baseline as well as postacetazolamide rCBF in the ipsilateral side (regression analysis, P < .05). Diameter and blood flow volume in the extracranial distal ICA were associated with ipsilateral postacetazolamide rCBF and VR (regression analysis, P < .05). When the patients were divided into 2 groups according to the ICA volume flow distal to a carotid stenosis, group I < 3.5 mL/s and group II > 3.5 mL/s, ipsilateral postacetazolamide rCBF in group I was significantly lower than that in group II (P = .008). Ipsilateral postacetazolamide rCBF (analysis of variance [ANOVA], P = .02) and VR (ANOVA, P = .03) significantly improved after CEA for 3 months in group I but not in group II. TOCU can detect the decrease in poststenotic flow of the distal extracranial ICA that is indicative of impaired intracranial hemodynamics and predictive for improvement of cerebral blood flow after CEA in patients with unilateral carotid stenosis.


Specification of neuronal and glial subtypes from human pluripotent stem cells.
Human pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), provide a dynamic tool for revealing early embryonic development, modeling pathological processes, and developing therapeutics through drug discovery and potential cell replacement. The first step toward the utilities of human PSCs is directed differentiation to functionally specialized cell/tissue types. Following developmental principles, human ESCs, and lately iPSCs, have been effectively differentiated to region- and/or transmitter-specific neuronal and glial types, including cerebral glutamatergic, striatal γ-aminobutyric acid (GABA)-ergic, forebrain cholinergic, midbrain dopaminergic, and spinal motor neurons, as well as astrocytes and oligodendrocytes. These studies also reveal unique aspects of human cell biology, including intrinsically programmed developmental course, differential uses of transcription factors for neuroectoderm specification, and distinct responses to extracellular signals in regulating cell fate. Such information will be instrumental in translating biological findings to therapeutic development.


Therapeutic progress--review VII. The medical treatment of Paget's disease.
The presentation of Paget's disease varies from a painful or deforming skeletal affliction to an asymptomatic disorder diagnosed on routine biochemical or radiological assessment. When involvement of the peripheral skeleton by Paget's disease is extensive, the clinical diagnosis is usually clear. Affected bones are thickened and deformed and the overlying skin is warm. Bone pain is sometimes severe and malignant change rarely occurs. The new bone formed is structurally abnormal and is consequently liable to deformity and fractures. Serum alkaline phosphatase concentrations and urinary hydroxyproline excretion are raised. Characteristic X-ray changes are seen. Paget's disease should be treated when it causes skeletal pain and tenderness, or when there are neurological symptoms, fractures, marked deformities, or other complications. New therapeutic agents offer both symptomatic relief and some control of the basic disease process. Simple analgesics should be tried before proceeding to the anti-osteoclastic agents, calcitonin, diphosphonates and mithramycin. All are effective in relieving bone pain and improving biochemical indices. The major advantage of the diphosphonates lies in their oral usage and thus, the number of patients who nowadays require calcitonin is small. The majority of patients should be commenced on a course of diphosphonate therapy (EHDP in most instances), but if clinical response is unsatisfactory calcitonin should be tried. Mithramycin should be reserved for special indications e.g. an elderly patient with severe disabling pain.


Intranuclear delivery of synthetic nuclear factor-kappa B p65 reduces inflammasomes after surgery.
Patients undergoing surgery can suffer from various complications, including post-operative bleeding, local or systematic infection, and neurologic disorders. Major surgery can initiate innate immune responses and trigger overproduction of inflammatory mediators, which can contribute to organ dysfunction. Inflammasomes are innate immune complexes, which are connected to the pathogenesis of various diseases, including atherosclerosis, hemorrhagic brain injury, and Alzheimer's disease. In the present study, we hypothesized that nucleotide-binding oligomerization domain-containing-like receptor protein (NLRP) inflammasomes may have a role in the pathological effects of surgery. Therefore, we designed a protein inhibitor of nuclear factor kappa B (NF-κB) p65 transcripts, called nt-p65-TMD (nuclear transducible (nt) transcription modulated domain (TMD) of RelA (p65)), that can penetrate the nucleus, and evaluated its therapeutic efficacy for dampening surgery-induced inflammasome activation. It was found that the nt-p65-TMD significantly reduced the NLRP1 inflammasome complex components (NLRP1, ASC, and Caspase-1) and interleukin (IL)-1β and IL-18 productions in the spleen after surgery. In the spleen, specific cell population and selective mediators were altered after surgery with/without nt-p65-TMD treatment. Also, we found that treatment of nt-p65-TMD decreased cell death in the spleen after surgery. Therefore, nt-p65-TMD is a potential novel strategy for reducing surgery-induced NLRP1 inflammasome and complications.


A comparison of the complement fixation, indirect fluorescent antibody, and microagglutination tests for the serological diagnosis of rickettsial diseases.
Three techniques for the serological diagnosis of Rocky Mountain spotted fever were compared by testing 417 sera from 178 patients who very probably did not have rickettsial infections and 88 sera from 41 patients who very probably had Rocky Mountain spotted fever (SF). The techniques were complement fixation (CF), indirect fluorescent antibody (IFA), and microagglutination (MA). To avoid possible degradation during unnecessary purification, the antigens were prepared by methods that were as simple as possible. In the CF tests of 417 sera from patients with nonrickettsial diseases there was only one titer of 8 and none at higher dilutions, whereas with the IFA and MA tests 4-8% of the sera reacted with SF antigens and 4-20% reacted with murine typhus (MT) antigens; the evidence indicated that these reactions were not caused by specific rickettsial antibody. With the SF sera, it could be seen that the IFA test was the most sensitive and the MA test was the least sensitive at each interval after infection. Moreover, the IFA results showed the least number of confusing cross-reactions with MT antigens and the MA test showed the most. The relative advantages of the three tests in serodiagnosis of rickettsial diseases are discussed.


Cytoplasmic, nuclear, membrane-bound and secreted [35S]methionine-labelled polypeptide pattern in differentiating fibroblast stem cells in vitro.
Methods for the selective enrichment of various subpopulations of the human skin fibroblast cell line HH-8 have been developed. These methods permit the selection of homogeneous populations of the three mitotic fibroblast cell types MF I, II and III, and the four postmitotic cell types PMF IV, V, VI and VII. These seven cell types exhibit differentiation-dependent and cell-type-specific patterns of [35S]methionine-labelled polypeptides in total soluble cytoplasmic and nuclear proteins, also in membrane-bound proteins, and in secreted proteins. In the differentiation sequence MF II-MF III-PMF IV - PMF V - PMF VI 14 cell-type-specific marker proteins have been found in the cytoplasmic and nuclear fraction, also 24 cell-type-specific marker proteins have been found in the membrane-bound protein fraction, and 11 cell-type-specific marker proteins in the secreted protein fraction. Markers in spontaneously arising and experimentally selected or induced populations of a single fibroblast cell type were found to be identical.


Biopolymer Patterning-Directed Secretion in Mucoid and Nonmucoid Strains of Pseudomonas aeruginosa Revealed by Multimodal Chemical Imaging.
Quinolone, pyocyanin, and rhamnolipid production were studied in Pseudomonas aeruginosa by spatially patterning mucin, a glycoprotein important to infection of lung epithelia. Mass spectrometric imaging and confocal Raman microscopy are combined to probe P. aeruginosa biofilms from mucoid and nonmucoid strains grown on lithographically defined patterns. Quinolone signatures from biofilms on patterned vs unpatterned and mucin vs mercaptoundecanoic acid (MUA) surfaces were compared. Microbial attachment is accompanied by secretion of 2-alkyl-4-quinolones as well as rhamnolipids from the mucoid and nonmucoid strains. Pyocyanin was also detected both in the biofilm and in the supernatant in the mucoid strain only. Significant differences in the spatiotemporal distributions of secreted factors are observed between strains and among different surface patterning conditions. The mucoid strain is sensitive to composition and patterning while the nonmucoid strain is not, and in promoting community development in the mucoid strain, nonpatterned surfaces are better than patterned, and mucin is better than MUA. Also, the mucoid strain secretes the virulence factor pyocyanin in a way that correlates with distress. A change in the relative abundance for two rhamnolipids is observed in the mucoid strain during exposure to mucin, whereas minimal variation is observed in the nonmucoid strain. Differences between mucoid and nonmucoid strains are consistent with their strain-specific phenology, in which the mucoid strain develops highly protected and withdrawn biofilms that achieve Pseudomonas quinolone signal production under limited conditions.


Diffusion-weighted MR imaging of solid and cystic lesions of the pancreas.
Diffusion-weighted magnetic resonance (MR) imaging is increasingly used in the detection and characterization of pancreatic lesions. Diffusion-weighted imaging may provide additional information to radiologists evaluating patients who have cystic or solid neoplasms of the pancreas. Because of greater freedom of motion of water molecules in fluid-rich environments, simple cysts in the pancreas have higher signal intensity on diffusion-weighted images with a b value of 0 sec/mm2 and lower signal intensity on high-b-value images. High apparent diffusion coefficient (ADC) values can be obtained on ADC maps because of the T2 "shine-through" effect. In contrast, solid neoplasms of the pancreas show increased signal intensity relative to the pancreas on diffusion-weighted images with a b value of 0 sec/mm2 and relatively high signal intensity on high-b-value images. Diffusion-weighted imaging can help detect solid pancreatic neoplasms with extremely dense cellularity or extracellular fibrosis by demonstrating significantly low ADC values, and these neoplasms may be better detected on diffusion-weighted MR images because of better contrast, although the resolution is generally worse. However, diffusion-weighted imaging may not be capable of helping definitively characterize solid lesions as inflammatory or neoplastic because of an overlap in ADC values between the two types. For example, it is difficult to distinguish poorly differentiated pancreatic adenocarcinoma from mass-forming pancreatitis at diffusion-weighted imaging because of similarly low ADC values attributed to dense fibrosis.


Chikungunya Virus: Pathophysiology, Mechanism, and Modeling.
Chikungunya virus (CHIKV), a mosquito-transmitted alphavirus, is recurring in epidemic waves. In the past decade and a half, the disease has resurged in several countries around the globe, with outbreaks becoming increasingly severe. Though CHIKV was first isolated in 1952, there remain significant gaps in knowledge of CHIKV biology, pathogenesis, transmission, and mechanism. Diagnosis is largely simplified and based on symptoms, while treatment is supportive rather than curative. Here we present an overview of the disease, the challenges that lie ahead for future research, and what directions current studies are headed towards, with emphasis on improvement of current animal models and potential use of 3D models.


microRNAs: short non-coding bullets of gain of function mutant p53 proteins.
TP53 gene mutations are present in more than half of all human cancers. The resulting proteins are mostly full-length with a single aminoacid change and are abundantly present in cancer cells. Some of mutant p53 proteins gain oncogenic activities through which actively contribute to the aberrant cell proliferation, increased resistance to apoptotic stimuli and ability to metastatize of cancer cells. Gain of function mutant p53 proteins can transcriptionally regulate the expression of a large plethora of target genes. This mainly occurs through the formation of oncogenic transcriptional competent complexes that include mutant p53 protein, known transcription factors, posttranslational modifiers and scaffold proteins. Mutant p53 protein can also transcriptionally regulate the expression of microRNAs, small non-coding RNAs that regulate gene expression at the posttranscriptional level. Each microRNA can putatively target the expression of hundred mRNAs and consequently impact on many cellular functions. Thus, gain of function mutant p53 proteins can exert their oncogenic activities through the modulation of both non-coding and coding regions of human genome.


Dance training is superior to repetitive physical exercise in inducing brain plasticity in the elderly.
Animal research indicates that a combination of physical activity and sensory enrichment has the largest and the only sustaining effect on adult neuroplasticity. Dancing has been suggested as a human homologue to this combined intervention as it poses demands on both physical and cognitive functions. For the present exploratory study, we designed an especially challenging dance program in which our elderly participants constantly had to learn novel and increasingly difficult choreographies. This six-month-long program was compared to conventional fitness training matched for intensity. An extensive pre/post-assessment was performed on the 38 participants (63-80 y), covering general cognition, attention, memory, postural and cardio-respiratory performance, neurotrophic factors and-most crucially-structural MRI using an exploratory analysis. For analysis of MRI data, a new method of voxel-based morphometry (VBM) designed specifically for pairwise longitudinal group comparisons was employed. Both interventions increased physical fitness to the same extent. Pronounced differences were seen in the effects on brain volumes: Dancing compared to conventional fitness activity led to larger volume increases in more brain areas, including the cingulate cortex, insula, corpus callosum and sensorimotor cortex. Only dancing was associated with an increase in plasma BDNF levels. Regarding cognition, both groups improved in attention and spatial memory, but no significant group differences emerged. The latter finding may indicate that cognitive benefits may develop later and after structural brain changes have taken place. The present results recommend our challenging dance program as an effective measure to counteract detrimental effects of aging on the brain.


Bedlam Community Health Clinic: a collaborative interdisciplinary health care service for the medically indigent.
To describe a collaborative interdisciplinary health care service delivery system for the medically indigent in the Tulsa and northeastern Oklahoma area independent of state or federal funding. Northeastern Oklahoma and Tulsa. Community and ambulatory care for indigent patients. Bedlam Community Health Clinic (BCHC), which opened in August 2003, provides services to patients through partnerships among the health care disciplines of the University of Oklahoma College of Pharmacy, Nursing, and Medicine; is staffed by community physicians and pharmacists who volunteer their time and expertise; and is funded by the local community. Experiences and patients served. BCHC is a collaborative interdisciplinary clinic that addresses the needs of the medically indigent in Tulsa and northeastern Oklahoma. Conceived, developed, and funded by the local community, it does not depend on state or federal funding. A variety of services, both general medicine and specialty, are provided through BCHC. Since its opening in August 2003, the clinic has provided hands-on training for students from a variety of health care disciplines. The pharmacist-patient encounters provide helpful, meaningful drug information. The participation of volunteer pharmacists enables medically underserved or indigent patients to access pharmaceutical care and addresses the diverse health care needs of this often overlooked population. Health professionals and students in the Tulsa area have created an innovative mechanism for serving indigent patients who otherwise would lack adequate health care services.


Method for the preparation of mucosal flaps from the jejunum of laying hens for transporter studies in Ussing chambers.
Ussing chambers are frequently used for in vitro evaluation of intestinal transport physiology. The current study describes investigating the jejunal tissue from laying hens using a specific preparation method and evaluates the effect of glutamine in the maintenance buffer. Tunica mucosa was stripped from 104 jejunal samples from 10 hens and stabilised by a net device. Fifty samples were maintained with modified Krebs-Henseleit buffer (Control), 54 samples with additional 5 mM glutamine (Group Gln). The percentage of responding samples varied between 87 and 100%. Mean short circuit current (ΔI sc,) [µA/cm(2)] of samples exposed to 10 mM glucose in the Control group and Group Gln was 17.0 and 14.6 (p = 0.836), respectively, of samples exposed to 100 µM phloridzin -13.3 and -11.8 (p = 0.712), respectively, and of samples exposed to 100 µM carbachol 4.7 and 3.7 (p = 0.450), respectively. In conclusion, the net-supported method enabled a reliable investigation of jejunum from laying hens. Glutamine in the maintenance buffer was of no significant benefit.


Integrating intimate partner violence assessment and intervention into healthcare in the United States: a systems approach.
The Institute of Medicine, United States Preventive Services Task Force (USPSTF), and national healthcare organizations recommend screening and counseling for intimate partner violence (IPV) within the US healthcare setting. The Affordable Care Act includes screening and brief counseling for IPV as part of required free preventive services for women. Thus, IPV screening and counseling must be implemented safely and effectively throughout the healthcare delivery system. Health professional education is one strategy for increasing screening and counseling in healthcare settings, but studies on improving screening and counseling for other health conditions highlight the critical role of making changes within the healthcare delivery system to drive desired improvements in clinician screening practices and health outcomes. This article outlines a systems approach to the implementation of IPV screening and counseling, with a focus on integrated health and advocacy service delivery to support identification and interventions, use of electronic health record (EHR) tools, and cross-sector partnerships. Practice and policy recommendations include (1) ensuring staff and clinician training in effective, client-centered IPV assessment that connects patients to support and services regardless of disclosure; (2) supporting enhancement of EHRs to prompt appropriate clinical care for IPV and facilitate capturing more detailed and standardized IPV data; and (3) integrating IPV care into quality and meaningful use measures. Research directions include studies across various health settings and populations, development of quality measures and patient-centered outcomes, and tests of multilevel approaches to improve the uptake and consistent implementation of evidence-informed IPV screening and counseling guidelines.


Linear constraint relations in biochemical reaction systems: I. Classification of the calculability and the balanceability of conversion rates.
Measurements provide the basis for process monitoring and control as well as for model development and validation. Systematic approaches to increase the accuracy and credibility of the empirical data set are therefore of great value. In (bio)chemical conversions, linear conservation relations such as the balance equations for charge, enthalpy, and/or chemical elements, can be employed to relate conversion rates. In a pactical situation, some of these rates will be measured (in effect, be calculated directly from primary measurements of, e.g., concentrations and flow rates), as others can or cannot be calculated from the measured ones. When certain measured rates can also be calculated from other measured rates, the set of equations, the accuracy and credibility of the measured rates can indeed be improved by, respectively, balancing and gross error diagnosis. The balanced conversion rates are more accurate, and form a consistent set of data, which is more suitable for further application (e.g., to calculate nonmeasured rates) than the raw measurements. Such an approach has drawn attention in previous studies. The current study deals mainly with the problem of mathematically classifying the conversion rates into balanceable and calculable rates, given the subset of measured rates. The significance of this problem is illustrated with some examples. It is shown that a simple matrix equation can be derived that contains the vector of measured conversion rates and the redundancy matrix R. Matrix R plays a predominant role in the classification problem. In supplementary articles, significance of the redundancy matrix R for an improved gross error diagnosis approach will be shown. In addition, efficient equations have been derived to calculate the balanceable and/or calculable rates. The method is completely based on matrix algebra (principally different from the graph-theoretical approach), and it is easily implemented into a computer program. (c) 1994 John Wiley & Sons, Inc.


Recent progress of light manipulation strategies in organic and perovskite solar cells.
Organic and perovskite solar cells are suffering from the insufficient utilization of incident light and thus low light harvesting efficiency despite their rapid progress in the past decade. In this regard, light manipulation strategies have attracted numerous attention to solve this inherent limit. Herein, the recent advances in light manipulation techniques in this area are overviewed. The light manipulation mechanisms are illustrated to classify the structures. Various light manipulation structures, fabrication techniques, and corresponding results are given and discussed, addressing the suppression of surface reflection, nano/micro-structure-induced light scattering, and the plasmonic effects with periodic metallic patterns and metallic nanoparticles. A brief perspective on future research is also proposed for pursuing broadband light harvesting.


Pancreatic carcinoma. Survival following detection by ultrasonic scanning.
The survival of 28 patients with pancreatic cancer was studied following initial detection of this malignant neoplasm by ultrasonic scanning. By ultrasound examination, 24 patients had an abnormality, and 21 had a demonstrable mass. Twenty-seven patients are now deceased, and the overall survival ranged from eight days to 24 months (mean, 6.1 months). The results of this study indicate that diagnosis by ultrasonic scanning had no effect on the survival of patients with pancreatic carcinoma.


Mucoidy, quorum sensing, mismatch repair and antibiotic resistance in Pseudomonas aeruginosa from cystic fibrosis chronic airways infections.
Survival of Pseudomonas aeruginosa in cystic fibrosis (CF) chronic infections is based on a genetic adaptation process consisting of mutations in specific genes, which can produce advantageous phenotypic switches and ensure its persistence in the lung. Among these, mutations inactivating the regulators MucA (alginate biosynthesis), LasR (quorum sensing) and MexZ (multidrug-efflux pump MexXY) are the most frequently observed, with those inactivating the DNA mismatch repair system (MRS) being also highly prevalent in P. aeruginosa CF isolates, leading to hypermutator phenotypes that could contribute to this adaptive mutagenesis by virtue of an increased mutation rate. Here, we characterized the mutations found in the mucA, lasR, mexZ and MRS genes in P. aeruginosa isolates obtained from Argentinean CF patients, and analyzed the potential association of mucA, lasR and mexZ mutagenesis with MRS-deficiency and antibiotic resistance. Thus, 38 isolates from 26 chronically infected CF patients were characterized for their phenotypic traits, PFGE genotypic patterns, mutations in the mucA, lasR, mexZ, mutS and mutL gene coding sequences and antibiotic resistance profiles. The most frequently mutated gene was mexZ (79%), followed by mucA (63%) and lasR (39%) as well as a high prevalence (42%) of hypermutators being observed due to loss-of-function mutations in mutL (60%) followed by mutS (40%). Interestingly, mutational spectra were particular to each gene, suggesting that several mechanisms are responsible for mutations during chronic infection. However, no link could be established between hypermutability and mutagenesis in mucA, lasR and mexZ, indicating that MRS-deficiency was not involved in the acquisition of these mutations. Finally, although inactivation of mucA, lasR and mexZ has been previously shown to confer resistance/tolerance to antibiotics, only mutations in MRS genes could be related to an antibiotic resistance increase. These results help to unravel the mutational dynamics that lead to the adaptation of P. aeruginosa to the CF lung.


Motoric cognitive risk syndrome trajectories and incident dementia over 10 years.
Motoric Cognitive Risk (MCR) syndrome is a high-risk state for adverse health outcomes in older adults characterised by measured slow gait speed and self-reported cognitive complaints. The recent addition to the Lothian Birth Cohort 1936 of robust dementia outcomes enabled us to assess the prognostic value of MCR for dementia and explore the various trajectories of participants diagnosed with MCR. We classified 680 community-dwelling participants free from dementia into non-MCR or MCR groups at mean [SD] age 76.3 [0.8] years. We used Cox and competing risk regression methods, adjusted for potential confounders, to evaluate the risk of developing all-cause incident dementia over 10 years of follow-up. Secondarily, we followed the trajectories for individuals with and without MCR at baseline and categorised them into subgroups based on whether MCR was still present at the next research wave, three years later. The presence of MCR increased the risk of incident dementia (adjusted HR 2.34, 95%CI 1.14-4.78, p = 0.020), as did fewer years of education and higher depression symptoms. However, MCR has a heterogenous progression trajectory. The MCR progression subgroups each have different prognostic values for incident dementia. MCR showed similar prognostic ability for dementia in a Scottish cohort as for other populations. MCR could identify a target group for early interventions of modifiable risk factors to prevent incident dementia. This study illustrates the heterogeneous nature of MCR progression. Exploring the underlying reasons will be important work in future work.


Estimation of the Prevalence of Amyotrophic Lateral Sclerosis in the United States Using National Administrative Healthcare Data from 2002 to 2004 and Capture-Recapture Methodology.
National administrative healthcare data may be used as a case-finding method for prevalence studies of chronic disease in the United States, but the completeness of ascertainment likely varies depending on the disease under study. We used 3 case-finding sources (Medicare, Medicaid, and Veterans Administration data) to estimate the prevalence of amyotrophic lateral sclerosis (ALS) in the United States for 2002-2004, and applied the capture-recapture methodology to estimate the degree of under-ascertainment when relying solely on these sources for case identification. Case-finding completeness was 76% overall and did not vary by race, but was lower for males (77%) than for females (88%), and lower for patients under age 65 (66%) than patients over age 65 (79%). The uncorrected ALS prevalence ratio was 2.8/100,000 in 2002, 3.3/100,000 in 2003, and 3.7/100,000 in 2004. After correcting for under-ascertainment, the annual prevalence increased by approximately 1 per 100,000 to 3.7/100,000 in 2002 (95% CI 3.66-3.80), 4.4/100,000 in 2003 (95% CI 4.34-4.50), and 4.8/100,000 in 2004 (95% CI 4.76-4.91). Federal healthcare claims databases ascertained are a very efficient method for identifying the majority of ALS-prevalent cases in the National ALS Registry, and may be enhanced by having patients self-register through the registry web portal.


A computational approach to mortality prediction of alcohol use disorder inpatients.
Health information technologies can assist clinicians in the Intensive Care Unit (ICU) by providing additional analysis of patient stability. However, because patient diagnoses can be confounded by chronic alcohol use, the predictive value of existing systems is suboptimal. Through the use of Electronic Health Records (EHR), we have developed computer software called AutoTriage to generate accurate predictions through multi-dimensional analysis of clinical variables. We analyze the performance of AutoTriage on the Alcohol Use Disorder (AUD) subpopulation in this study, and build on results we reported for AutoTriage performance on the general population in previous work. AUD-related ICD-9 codes were used to obtain a patient population from MIMIC III ICU dataset for a retrospective study. Patient mortality risk score is generated through analysis of eight EHR-based clinical variables. The score is determined by combining weighted subscores, each of which are obtained from singlets, doublets or triplets of one or more of the eight continuous-valued clinical variable inputs. A temporally updating risk score is computed with a continuously revised 12-hour mortality prediction. Among AUD patients, in a non-overlapping test set, AutoTriage outperforms existing systems with an Area Under Receiver Operating Characteristic (AUROC) value of 0.934 for 12-h mortality prediction. At a sensitivity of 90%, AutoTriage achieves a specificity of 80%, positive predictive value of 40%, negative predictive value of 89%, and an Odds Ratio of 36. For mortality prediction, AutoTriage demonstrates improvements in both the accuracy and the Odds Ratio over current systems among the AUD patient population.


Na+ transport by mammalian stomach.
Gastric mucosas from newborn pigs (0--20 days) and rabbits (0--20 days) were used for in vitro investigation of active Na+ transport during resting (no HCl secretion) conditions. As measured with 22Na+, these tissues actively absorb Na+ from the mucosal-to serosal (m-t-s) bathing solution during both open-circuit and short-circuit current (Is) conditions. In the nonsecreting state, net Na+ transport accounts for 40--60% of Isc. The remaining current is provided by net s-to-m flux of Cl-. Amiloride (2-5 X 10(-5) M) in the mucosal solution abolishes this active Na+ transport by inhibiting m-to-s fluxes of Na+ (JNams). In vivo-in vitro experiments showed that active Na+ transport is a normal function of the resting mammalian stomach. Decreasing pH of the mucosal solution below pH 5 reversibly causes decreased current-generating capability of the tissue. Pretreatment of the tissue with amiloride abolishes this pH effect. The implication is that the low pH affects the Na+-entry step into cells. "Titration curves" of current vs. pH had an apparent pK approximately 4.0. Ouabain and K+-free solutions both cause decreases in active Na+ and Cl- current. Calculations indicate that a shunt may account for only a small (less than 30%) percentage of total transepithelial conductance.


Balanced production of 30S and 50S ribosomal proteins after a nutritional shift-up.
The synthesis of bulk ribosomal protein (r-protein) after a nutritional shift-up in Escherichia coli B/r was examined. It was found that the molar ratio of the net synthesis rates of 30S and 50S r-protein remains constant during the transition period after the shift-up and equal to the preshift ratio. The implications for the control of ribosome synthesis are discussed.


[Role of lactose, arginine and lysine combination in fracture healing (an experimental study)].
L-arginine and L-lysine are essential amino acids which seem to possess some properties able to influence bone fractures healing. In fact, they seem to be involved not only in the increase of intestinal calcium adsorption but also in collagen synthesis, in insulin and growth hormone secretion and in osteoblastic activation. So, an experimental in vivo model was carried out by using 50 adult rabbits which, under general anaesthesia, were submitted to an osteotomy of the left fibula. Animals were divided into 5 groups and were daily treated with a mixture of lactose, L-arginine and L-lysine or with the only lactose (control group) at the same dosage as recommended for humans. They were sacrificed after 15, 30, 40, 50 and 60 days for radiological and histological studies. The results of the study showed that the pharmacological mixture containing L-arginine and L-lysine accelerates and ameliorates the healing processes and this positive effect was particularly evident from the 30th day after the osteotomy. We think that these results are linked not only to calcium metabolism but also to different biological properties which positively contribute to a good healing of bone fractures.


Electron-microscopic and electrophoretic studies of bovine femoral-head cartilage proteoglycan fractions.
Proteoglycans (A1D1) extracted from bovine femoral-head cartilage were examined by electron microscopy using benzyldimethylammonium chloride as a spreading agent. The preparation contained a mixture of particles, some with a 'beaded' structure and a contiguous filamentous 'tail' at one end and others which appeared as round 'blobs', some of which also had filamentous tails. Previous electron-microscopic studies of proteoglycan monomers have indicated that their length distributions were apparently unimodal, a finding that contrasted with agarose/polyacrylamide-gel-electrophoresis results, which generally indicated two bands. In the present study proteoglycans isolated from the slowly migrating electrophoretic band were shown to be predominantly the larger molecules of beaded appearance, whereas the rapidly migrating proteoglycans were predominantly molecules with the 'blob-like' appearance. Gel-filtration, isopycnic-density-gradient-centrifugation and rate-zonal-centrifugation techniques were evaluated as means of proteoglycan fractionation by electron microscopy and agarose-gel electrophoresis. Rate-zonal centrifugation in mixed-salt gradients of caesium chloride/4 M-guanidinium chloride yielded the most effective fractionation.


Bridging the gap: increasing collaboration between research mentors and career development educators for PhD and postdoctoral training success.
National reports and funding mandates have called for trainee-centered PhD and postdoctoral training and the need to support diverse career outcomes. As a result, career and professional development (CPD) resources have expanded at several institutions. Despite the growth of innovative and impactful CPD resources, access to and awareness of resources have been inconsistent and inequitable for graduate and postdoctoral trainees. In the current model, core education occurs in two unconnected ways: faculty research mentors provide scientific competencies training, while CPD educators provide transferable competencies training, which is separate from the curriculum and optional at most institutions. Research mentors are influential in supporting trainee engagement with CPD programs; however, most are either unaware of the rapidly growing opportunities or may not see the direct benefit to scientific development and productivity. Due to this disconnect, some trainees can be inadvertently distanced from CPD resources, leading to more inequities among groups. To bridge this gap, here we propose a realignment of the current model via a set of practical and collaborative solutions providing benefit to all stakeholders. With greater awareness and collaboration, research mentors and CPD educators can complement each other's expertise to better support trainee experiences and outcomes.


[Radiodiagnosis of the reparative osteogenesis during the treatment of comminuted fractures of the humerus].
Roentgenoradiological control of a reparative process after diaphysial comminuted shoulder fractures was exercised at all stages of the treatment after G. A. Ilizarov's method. Osteoscanning permitted the assessment of the viability of bone splinters on the first days after trauma and quantitative assessment of the activity of osteogenesis. A study of the optical density of radiograms provided an opportunity for evaluation of mineral content in regenerating bones, their structure only at late stages of treatment when it was rather high.


Reassessing the role of high dose cytarabine and mitoxantrone in relapsed/refractory acute myeloid leukemia.
A substantial segment of patients with acute myeloid leukemia (AML) will relapse following an initial response to induction therapy or will prove to be primary refractory. High-dose cytarabine and mitoxantrone (HiDAC/MITO) is an established salvage therapy for these patients. We studied all adult patients with relapsed/refractory (R/R) AML who were treated with HiDAC/MITO in our center between the years 2008-2017. To determine whether responding patients harbored a unique molecular signature, we performed targeted next-generation sequencing (NGS) on a subset of patients. The study cohort consisted of 172 patients with a median age of 54 years (range 18-77). The composite complete remission rate was 58%; 11 patients (6%) died during salvage therapy. Median survival was 11.4 months with a 1-year survival rate of 48%. In multivariate analysis favorable risk cytogenetics [Odds ratio (OR)=0.34, confidence interval (CI) 95%, 0.17-0.68; P = 0.002], and de-novo AML (OR = 0.4, CI 95%, 0.16-0.98; P = 0.047) were independently associated with a favorable response. Patients who attained a complete remission had a median survival of 43.7 months compared with 5.2 months for refractory patients (p < 0.0001). Neither the FLT3-ITD and NPM1 mutational status nor the indication for salvage therapy significantly impacted on the response to HiDAC/MITO salvage. NGS analysis identified 20 different mutations across the myeloid gene spectrum with a distinct TP53 signature detected in non-responding patients. HiDAC/MITO is an effective salvage regimen in R/R AML, however patients with adverse cytogenetics or secondary disease may not benefit as much from this approach.


Hierarchical, Self-Healing and Superhydrophobic Zirconium Phosphate Hybrid Membrane Based on the Interfacial Crystal Growth of Lyotropic Two-Dimensional Nanoplatelets.
We demonstrate a facile route to in situ growth of lyotropic zirconium phosphate (ZrP) nanoplates on textiles via an interfacial crystal growing process. The as-prepared hybrid membrane shows a hierarchical architecture of textile fibers (porous platform for fluid transport), ZrP nanoplatelets (layered scaffolds for chemical barriers), and octadecylamine (organic species for superhydrophobic functionalization). Interestingly, such a hybrid membrane is able to separate the oily wastewater with a high separation efficiency of 99.9%, even at in harsh environments. After being chemically etched, the hybrid membrane is able to restore its hydrophobicity autonomously and repeatedly, owing to the hierarchical structure that enables facile loading of healing agent. We anticipate that the concept of implanting superhydrophobic self-healing features in anisotropic structure of lyotropic nanoparticles will open up new opportunities for developing advanced multifunctional materials for wastewater treatment, fuel purification, and oil spill mitigation.


What predicts legislative success of early care and education policies?: Applications of machine learning and Natural Language Processing in a cross-state early childhood policy analysis.
Following the pioneering efforts of a federal Head Start program, U.S. state policymakers have rapidly expanded access to Early Care and Education (ECE) programs with strong bipartisan support. Within the past decade the enrollment of 4 year-olds has roughly doubled in state-funded preschool. Despite these public investments, the content and priorities of early childhood legislation-enacted and failed-have rarely been examined. This study integrates perspectives from public policy, political science, developmental science, and machine learning in examining state ECE bills in identifying key factors associated with legislative success. Drawing from the Early Care and Education Bill Tracking Database, we employed Latent Dirichlet Allocation (LDA), a statistical topic identification model, to examine 2,396 ECE bills across the 50 U.S. states during the 2015-2018. First, a six-topic solution demonstrated the strongest fit theoretically and empirically suggesting two meta policy priorities: 'ECE finance' and 'ECE services'. 'ECE finance' comprised three dimensions: (1) Revenues, (2) Expenditures, and (3) Fiscal Governance. 'ECE services' also included three dimensions: (1) PreK, (2) Child Care, and (3) Health and Human Services (HHS). Further, we found that bills covering a higher proportion of HHS, Fiscal Governance, or Expenditures were more likely to pass into law relative to bills focusing largely on PreK, Child Care, and Revenues. Additionally, legislative effectiveness of the bill's primary sponsor was a strong predictor of legislative success, and further moderated the relation between bill content and passage. Highly effective legislators who had previously passed five or more bills had an extremely high probability of introducing a legislation that successfully passed regardless of topic. Legislation with expenditures as policy priorities benefitted the most from having an effective legislator. We conclude with a discussion of the empirical findings within the broader context of early childhood policy literature and suggest implications for future research and policy.


[Long-term management of patients with chronic renal failure].
Any type of chronic renal disease is associated with functional deterioration of the kidney due to progressive glomerulosclerosis with interstitial fibrosis and tubular atrophy. This process is thought to be predominantly due either to glomerular hyperfiltration or mesangial overload with macromolecules. Antihypertensive therapy, particularly with ACE inhibitors, and protein restriction have been found to retard progressive glomerulosclerosis in animal experiments. There is no doubt that patients with renal disease benefit from antihypertensive therapy through both preservation of renal function and prevention of secondary organ damage due to hypertension. However, the value of protein restricted diets with or without supplements of essential amino acids or ketoacids is less clear. A patient treated with protein restriction is presented and the investigations necessary to monitor compliance, renal function and nutrition are discussed. Monthly to quarterly controls of renal function, blood pressure and mineral metabolism are suggested, particularly in the case of severe hypertension and of prophylactic treatment for renal osteodystrophy with phosphate binders and vitamin D metabolites. Finally, guidelines are provided for planning of renal replacement therapy by dialysis and renal transplantation in the individual patient.


Lack of focus on nutrition and documentation in nursing homes, home care- and home nursing: the self-perceived views of the primary care workforce.
Malnutrition is a comprehensive challenge for the nursing home, home care- and home nursing sector. Nutritional care and the subsequent documentation are a common and multifaceted healthcare practice that requires that the healthcare professionals possess complex combinations of competencies in order to deliver high-quality care and treatment. The purpose of this study was to investigate how a varied group of healthcare professionals' perceive their own competencies within nutrition and documentation and how organizational structures influence their daily work and the quality of care provided. Two focus groups consisting of 14 healthcare professionals were conducted. The transcribed focus group interviews was analyzed using the qualitative content analysis approach. Six categories were identified: 1) Lack of uniform and systematic communication affect nutritional care practices 2) Experience-based knowledge among the primary workforce influences daily clinical decisions, 3) Different attitudes towards nutritional care lead to differences in the quality of care 4) Differences in organizational culture affect quality of care, 5) Lack of clear nutritional care responsibilities affect how daily care is performed and 6) Lack of clinical leadership and priorities makes nutritional care invisible. The six categories revealed two explanatory themes: 1) Absent inter- and intra-professional collaboration and communication obstructs optimal clinical decision-making and 2) quality deterioration due to poorly-established nutritional care structure. Overall, the two themes explain that from the healthcare professionals' point of view, a visible organization that allocates resources as well as prioritizing and articulating the need for daily nutritional care and documentation is a prerequisite for high-quality care and treatment. Furthermore, optimal clinical decision making among the healthcare professionals are compromised by imprecise and unclear language and terminology in the patients' healthcare records and also a lack of clinical guidelines and standards for collaboration between different healthcare professionals working in nursing homes, home care or home nursing. The findings of this study are beneficial to support organizations within these settings with strategies focusing on increasing nutritional care and documentation competencies among the healthcare professionals. Furthermore, the results advocate for the daily involvement and support of leaders and managers in articulating and structuring the importance of nutritional care and treatment and the subsequent documentation.


Degeneration of nigrostriatal dopaminergic neurons increases iron within the substantia nigra: a histochemical and neurochemical study.
Parkinson's-diseased (PD) brains have increased levels of iron in the zona compacta of the substantia nigra (SNc). To determine whether these elevated nigral iron levels may be caused secondarily by degeneration of nigrostriatal dopaminergic (NS-DA) neurons, the NS-DA pathway was unilaterally lesioned in rats through 6-hydroxydopamine (6-OHDA) infusion and nigral iron levels evaluated three weeks later. A significant increase was observed in both iron concentration (+35%) and iron content (+33) within the substantia nigra (SN) ipsilateral to comprehensive 6-OHDA lesions. Moreover, ferric iron staining was dramatically increased within the SNc following 6-OHDA lesions, primarily due to the appearance of iron-positive SNc neurons and infiltrating reactive glial cells. Iron staining in the SN zona reticularis was modestly increased after 6-OHDA lesions, but staining in the neostriatum and globus pallidus was unaffected. These results indicate that loss of NS-DA neurons is associated with increased iron levels in the SN. This suggests that increased nigral iron levels in PD may be secondary to some neurodegenerative process. Nonetheless, even a secondary increase in nigral iron levels may be of pathogenic importance in PD because of iron's ability to catalyze neurotoxic free radical formation and perpetuate neurodegeneration.


Isolation of a Pseudomonas sp. Which Utilizes the Phosphonate Herbicide Glyphosate.
A strain of bacteria has been isolated which rapidly and efficiently utilizes the herbicide glyphosate (N-phosphonomethylglycine) as its sole phosphorus source in a synthetic medium. The strain (PG2982) was isolated by subculturing Pseudomonas aeruginosa ATCC 9027 in a synthetic broth medium containing glyphosate as the sole phosphorus source. Strain PG2982 differs from the culture of P. aeruginosa in that it is nonflagellated, does not produce pyocyanin, and has an absolute requirement for thiamine. Strain PG2982 has been tentatively identified as a Pseudomonas sp. strain by its biochemical activities and moles percent guanine plus cytosine. Measurements of glyphosate with an amino acid analyzer show that glyphosate rapidly disappears from the medium during exponential growth of strain PG2982. In batch culture at 30 degrees C, this isolate completely utilized 1.0 mM glyphosate in 96 h and yielded a cell density equal to that obtained with 1.0 mM phosphate as the phosphorus source. However, a longer lag phase and greater generation time were noted in the glyphosate-containing medium. Strain PG2982 can efficiently utilize glyphosate as an alternate phosphorus source.


System for High-Intensity Evaluation During Radiation Therapy (SHIELD-RT): A Prospective Randomized Study of Machine Learning-Directed Clinical Evaluations During Radiation and Chemoradiation.
Patients undergoing outpatient radiotherapy (RT) or chemoradiation (CRT) frequently require acute care (emergency department evaluation or hospitalization). Machine learning (ML) may guide interventions to reduce this risk. There are limited prospective studies investigating the clinical impact of ML in health care. The objective of this study was to determine whether ML can identify high-risk patients and direct mandatory twice-weekly clinical evaluation to reduce acute care visits during treatment. During this single-institution randomized quality improvement study (ClinicalTrials.gov identifier: NCT04277650), 963 outpatient adult courses of RT and CRT started from January 7 to June 30, 2019, were evaluated by an ML algorithm. Among these, 311 courses identified by ML as high risk (> 10% risk of acute care during treatment) were randomized to standard once-weekly clinical evaluation (n = 157) or mandatory twice-weekly evaluation (n = 154). Both arms allowed additional evaluations on the basis of clinician discretion. The primary end point was the rate of acute care visits during RT. Model performance was evaluated using receiver operating characteristic area under the curve (AUC) and decile calibration plots. Twice-weekly evaluation reduced rates of acute care during treatment from 22.3% to 12.3% (difference, -10.0%; 95% CI, -18.3 to -1.6; relative risk, 0.556; 95% CI, 0.332 to 0.924; P = .02). Low-risk patients had a 2.7% acute care rate. Model discrimination was good in high- and low-risk patients undergoing standard once-weekly evaluation (AUC, 0.851). In this prospective randomized study, ML accurately triaged patients undergoing RT and CRT, directing clinical management with reduced acute care rates versus standard of care. This prospective study demonstrates the potential benefit of ML in health care and offers opportunities to enhance care quality and reduce health care costs.


The Huntington's Disease Register of Papua New Guinea.
The discovery of Huntington's chorea in New Britain, Papua New Guinea in 1977 was followed by the establishment of a National Huntington's Disease Register. At present there are 11 choreics and 136 potential choreics registered within 5 kindreds. The disease presents early in Papua New Guinea and the average age of onset in living choreics in New Britain was 21 years. Affected persons here do not as a rule seek medical advice, and an active case detection policy was therefore adopted. The development of a genetic register in an unsophisticated and superstitious community poses special problems. Genetic counselling has so far proved to be ineffective.


Correlation of SPECT imaging, biochemical parameters and mutation with systolic dysfunction.
We investigated systolic dysfunction by the use of biochemical laboratory tests and perfusion single-photon emission computed tomography imaging in 32 Pakistani subjects exhibiting symptoms of this disorder. To investigate underlying genetic causes, such as familial hypercholesterolemia, DNA samples from these subjects were screened by PCR-SSCP and DNA sequencing to detect changes in the low density lipoprotein receptor gene (LDLR). A novel mutation (1171G>A) in exon 8 and two polymorphisms (1167G>A and 1413 A>G) in exons 8 and 10 of the LDLR gene were found. In silico tools such as SIFT, PolyPhen-2, KD4v, and Project HOPE were used to predict the effect of this mutation on protein structure and function.


Antisense Therapy Attenuates Phospholamban p.(Arg14del) Cardiomyopathy in Mice and Reverses Protein Aggregation.
Inherited cardiomyopathy caused by the p.(Arg14del) pathogenic variant of the phospholamban (PLN) gene is characterized by intracardiomyocyte PLN aggregation and can lead to severe dilated cardiomyopathy. We recently reported that pre-emptive depletion of PLN attenuated heart failure (HF) in several cardiomyopathy models. Here, we investigated if administration of a Pln-targeting antisense oligonucleotide (ASO) could halt or reverse disease progression in mice with advanced PLN-R14del cardiomyopathy. To this aim, homozygous PLN-R14del (PLN-R14 Δ/Δ) mice received PLN-ASO injections starting at 5 or 6 weeks of age, in the presence of moderate or severe HF, respectively. Mice were monitored for another 4 months with echocardiographic analyses at several timepoints, after which cardiac tissues were examined for pathological remodeling. We found that vehicle-treated PLN-R14 Δ/Δ mice continued to develop severe HF, and reached a humane endpoint at 8.1 ± 0.5 weeks of age. Both early and late PLN-ASO administration halted further cardiac remodeling and dysfunction shortly after treatment start, resulting in a life span extension to at least 22 weeks of age. Earlier treatment initiation halted disease development sooner, resulting in better heart function and less remodeling at the study endpoint. PLN-ASO treatment almost completely eliminated PLN aggregates, and normalized levels of autophagic proteins. In conclusion, these findings indicate that PLN-ASO therapy may have beneficial outcomes in PLN-R14del cardiomyopathy when administered after disease onset. Although existing tissue damage was not reversed, further cardiomyopathy progression was stopped, and PLN aggregates were resolved.


Costimulatory ligand 4-1BBL (CD137L) as an efficient adjuvant for human antiviral cytotoxic T cell responses.
Effective adjuvants capable of inducing strong cytotoxic T cell responses in humans are lacking. In this study, we tested 4-1BBL as an adjuvant for activation of human memory antiviral CD8 T cell responses ex vivo. A recombinant replication-defective 4-1BBL adenovirus was used to convert autologous monocytes into efficient antigen-presenting cells after overnight incubation, bypassing the need to generate dendritic cells. Together with viral peptides, 4-1BBL led to robust memory responses of human Epstein-Barr virus- and influenza virus-specific cytotoxic T cells, with expansion of peptide-specific CD8 effector cells; up-regulation of Bcl-x(L), granzyme A, and perforin; enhanced cytotoxic activity; and increased cytokine production. The response was significant even at a 100-fold lower peptide dose, compared with responses obtained with control adenovirus. Adenovirus-delivered B7.1 also expanded and activated virus-specific CD8 T cells, but 4-1BBL was more effective in driving the T cells toward a more fully differentiated CD27(-) effector state. Thus, 4-1BBL is a promising adjuvant for human memory CD8 T cells and will likely be most effective in the boost phase of a prime-boost strategy.