Role of green technology innovation and renewable energy in carbon neutrality: A sustainable investigation from Turkey.
After the Paris Climate Conference (COP21), many countries start progressing towards carbon neutrality targets. In doing so, green technology innovations (GTIs) and clean energy are the essential factors that can help to achieve the carbon neutrality goal. Therefore, this paper examines the linkages between green technology innovation and renewable energy and carbon dioxide emissions based on the STIRPAT model in Turkey during the time of 1990-2018. The study used testing like "unit-root" to verify the variables' integrative properties containing the information for structural breaks. Also, the bootstrapping ARDL-bound testing technique is used to analyze the relationship between the variables. The causal relationship between green technology innovation, energy consumption, renewable energy, population, income per capita, and carbon dioxide emissions is tested through a Granger causality test. The empirical findings show that green technology innovation, renewable energy, energy consumption, population, income per capita, and carbon dioxide emissions are co-integrated for the long-term association. Additionally, green technology innovation and renewable energy decline carbon dioxide emissions, whereas energy consumption, population, and per capita enhance carbon emissions. This paper helps the policymakers design a comprehensive policy for strengthening environmental sustainability through green technology innovation and renewable energy, specifically in the region of Turkey.


Influence of powder composition and morphology on penetration of Gray and White ProRoot mineral trioxide aggregate and calcium hydroxide into dentin tubules.
This study examined the influence of powder composition and morphology on the penetration of Gray and White ProRoot mineral trioxide aggregate (GMTA, WMTA) and calcium hydroxide (CH) into open dentin tubules. GMTA, WMTA, and CH particle dimensions were analyzed by flow particle image analysis (FPIA). Penetration of open dentin tubules into dentin discs was studied by scanning electron microscopy. Five samples of each material were randomly selected and prepared for this study. The GMTA averages for length (μm), width (μm), perimeter (μm), and aspect ratio were 1.94 ± 1.65, 1.43 ± 1.19, 5.61 ± 4.27, and 0.76 ± 0.14, respectively. Corresponding averages for WMTA were 2.04 ± 1.87, 1.49 ± 1.33, 5.88 ± 4.81, and 0.76 ± 0.14, and for CH were 2.26 ± 1.99, 1.62 ± 1.46, 6.70 ± 5.60, and 0.74 ± 0.15, respectively. The rank order of the averages for particle length, width and perimeter from the largest to the smallest material was CH > WMTA > GMTA. The rank order of the averaged aspect ratios was GMTA > WMTA > CH. SEM showed that all three materials, when deposited and agitated on dentin discs, penetrated the open dentin tubules. Tubule occlusion occurred as particle surface concentrations increased. Significant differences in particle length, width, perimeter, and aspect ratio were observed for GMTA, WMTA, and CH (P < 0.0001 in all cases). All particle types penetrated into open tubules when agitated on dentin discs; all tubules were eventually occluded as particle concentrations grew. (J Oral Sci 56, 287-293, 2014).


A T cell-inducing influenza vaccine for the elderly: safety and immunogenicity of MVA-NP+M1 in adults aged over 50 years.
Current influenza vaccines have reduced immunogenicity and are of uncertain efficacy in older adults. We assessed the safety and immunogenicity of MVA-NP+M1, a viral-vectored influenza vaccine designed to boost memory T cell responses, in a group of older adults. Thirty volunteers (aged 50-85) received a single intramuscular injection of MVA-NP+M1 at a dose of 1·5×10(8) plaque forming units (pfu). Safety and immunogenicity were assessed over a period of one year. The frequency of T cells specific for nucleoprotein (NP) and matrix protein 1 (M1) was determined by interferon-gamma (IFN-γ) ELISpot, and their phenotypic and functional properties were characterized by polychromatic flow cytometry. In a subset of M1-specific CD8(+) T cells, T cell receptor (TCR) gene expression was evaluated using an unbiased molecular approach. Vaccination with MVA-NP+M1 was well tolerated. ELISpot responses were boosted significantly above baseline following vaccination. Increases were detected in both CD4(+) and CD8(+) T cell subsets. Clonality studies indicated that MVA-NP+M1 expanded pre-existing memory CD8(+) T cells, which displayed a predominant CD27(+)CD45RO(+)CD57(-)CCR7(-) phenotype both before and after vaccination. MVA-NP+M1 is safe and immunogenic in older adults. Unlike seasonal influenza vaccination, the immune responses generated by MVA-NP+M1 are similar between younger and older individuals. A T cell-inducing vaccine such as MVA-NP+M1 may therefore provide a way to circumvent the immunosenescence that impairs routine influenza vaccination. ClinicalTrials.gov NCT00942071.


Clinical evaluation of a single reaction, diagnostic polymerase chain reaction assay for the detection of hepatitis C virus RNA.
In the past few years the detection of HCV-RNA by polymerase chain reaction has become a well-established diagnostic tool for patients with chronic hepatitis C. However, the lack of reproducible results between laboratories and the relatively high proportion of false-positive results, has indicated the need for a standardized and reliable polymerase chain reaction assay. In the present study we have analyzed the performance of a commercial, HCV-RNA polymerase chain reaction assay based on a single, combined reverse transcription and amplification reaction and on the use of Uracil-N-glycosilase to prevent carry-over contamination (Amplicor HCV, Roche Molecular Systems). In this assay the amplification products are detected in microwell plates using biotinylated primer and the HRP avidin colorimetric system. Serum samples collected from 446 patients, including 181 with chronic active hepatitis C, 50 with autoimmune chronic hepatitis, 117 in hemodialysis, 30 asymptomatic carriers of anti-HCV and 68 with indeterminate serology (RIBA indeterminate results), as well as from 121 controls were tested with the commercial, single-step assay and with nested polymerase chain reaction. Both techniques use primers located within the 5' non-coding region of the HCV genome. In all cases a good concordance was observed between the commercial, single-step assay and nested polymerase chain reaction which, for patients with chronic active hepatitis, showed a sensitivity and specificity of 100% and 99.3% for the former and of 98.8% and 100% for the latter, when compared to clinical diagnosis taken as the gold standard. Most of the 11 discordant samples were seen in the group of RIBA-indeterminate cases and in patients with chronic active hepatitis C. Further analysis of these cases, based on repeat testing and clinical data showed that 64% and 36% of the discrepancies were due, respectively, to nested polymerase chain reaction and Amplicor inconsistent reactions. In hemodialyzed patients, patients with autoimmune hepatitis and asymptomatic carriers of anti-HCV, both assays produced results which were consistent with the clinical diagnosis. In the former group, polymerase chain reaction was able to identify the presence of active viral replication in some antibody negative samples. Taken together, these results indicate that the commercial, single-step polymerase chain reaction assay has the same clinical sensitivity and specificity as nested polymerase chain reaction and that, because of its simplified procedures and fast turn-around time, it may be a valuable test for routine diagnostic applications.


Cholinergic neurons of mouse intrinsic cardiac ganglia contain noradrenergic enzymes, norepinephrine transporters, and the neurotrophin receptors tropomyosin-related kinase A and p75.
Half of the cholinergic neurons of human and primate intrinsic cardiac ganglia (ICG) have a dual cholinergic/noradrenergic phenotype. Likewise, a large subpopulation of cholinergic neurons of the mouse heart expresses enzymes needed for synthesis of norepinephrine (NE), but they lack the vesicular monoamine transporter type 2 (VMAT2) required for catecholamine storage. In the present study, we determined the full scope of noradrenergic properties (i.e. synthetic enzymes and transporters) expressed by cholinergic neurons of mouse ICG, estimated the relative abundance of neurons expressing different elements of the noradrenergic phenotype, and evaluated the colocalization of cholinergic and noradrenergic markers in atrial nerve fibers. Stellate ganglia were used as a positive control for noradrenergic markers. Using fluorescence immunohistochemistry and confocal microscopy, we found that about 30% of cholinergic cell bodies contained tyrosine hydroxylase (TH), including the activated form that is phosphorylated at Ser-40 (pSer40 TH). Dopamine beta-hydroxylase (DBH) and norepinephrine transporter (NET) were present in all cholinergic somata, indicating a wider capability for dopamine metabolism and catecholamine uptake. Yet, cholinergic somata lacked VMAT2, precluding the potential for NE storage and vesicular release. In contrast to cholinergic somata, cardiac nerve fibers rarely showed colocalization of cholinergic and noradrenergic markers. Instead, these labels were closely apposed but clearly distinct from each other. Since cholinergic somata expressed several noradrenergic proteins, we questioned whether these neurons might also contain trophic factor receptors typical of noradrenergic neurons. Indeed, we found that all cholinergic cell bodies of mouse ICG, like noradrenergic cell bodies of the stellate ganglia, contained both tropomyosin-related kinase A (TrkA) and p75 neurotrophin receptors. Collectively, these findings demonstrate that mouse intrinsic cardiac neurons (ICNs), like those of humans, have a complex neurochemical phenotype that goes beyond the classical view of cardiac parasympathetic neurons. They also suggest that neurotrophins and local NE synthesis might have important effects on neurons of the mouse ICG.


Glycan-Gold Nanoparticles as Multifunctional Probes for Multivalent Lectin-Carbohydrate Binding: Implications for Blocking Virus Infection and Nanoparticle Assembly.
Multivalent lectin-glycan interactions are widespread in biology and are often exploited by pathogens to bind and infect host cells. Glycoconjugates can block such interactions and thereby prevent infection. The inhibition potency strongly depends on matching the spatial arrangement between the multivalent binding partners. However, the structural details of some key lectins remain unknown and different lectins may exhibit overlapping glycan specificity. This makes it difficult to design a glycoconjugate that can potently and specifically target a particular multimeric lectin for therapeutic interventions, especially under the challenging in vivo conditions. Conventional techniques such as surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC) can provide quantitative binding thermodynamics and kinetics. However, they cannot reveal key structural information, e.g., lectin's binding site orientation, binding mode, and interbinding site spacing, which are critical to design specific multivalent inhibitors. Herein we report that gold nanoparticles (GNPs) displaying a dense layer of simple glycans are powerful mechanistic probes for multivalent lectin-glycan interactions. They can not only quantify the GNP-glycan-lectin binding affinities via a new fluorescence quenching method, but also reveal drastically different affinity enhancing mechanisms between two closely related tetrameric lectins, DC-SIGN (simultaneous binding to one GNP) and DC-SIGNR (intercross-linking with multiple GNPs), via a combined hydrodynamic size and electron microscopy analysis. Moreover, a new term, potential of assembly formation (PAF), has been proposed to successfully predict the assembly outcomes based on the binding mode between GNP-glycans and lectins. Finally, the GNP-glycans can potently and completely inhibit DC-SIGN-mediated augmentation of Ebola virus glycoprotein-driven cell entry (with IC50 values down to 95 pM), but only partially block DC-SIGNR-mediated virus infection. Our results suggest that the ability of a glycoconjugate to simultaneously block all binding sites of a target lectin is key to robust inhibition of viral infection.


Efficacy of honey in comparison to topical corticosteroid for treatment of recurrent minor aphthous ulceration: a randomized, blind, controlled, parallel, double-center clinical trial.
Recurrent aphthous ulceration represents a very common mucosal disorder that general dentists may encounter on a daily basis, and for which there is no curative treatment. The best treatment that can be achieved is to avoid local traumatic precipitants, lessen the pain and duration of ulceration by suppressing the local immune response, and prevent secondary infection. The objective of this study was to clinically determine the efficacy of honey as a topical treatment of recurrent minor aphthous ulceration in a Saudi cohort. A randomized, blind, controlled, parallel, double-center clinical trial was carried out. Honey was applied by patients four times a day for 5 days. Clinical parameters (ulcer size, pain scale, and degree of erythema and healing) were recorded both at baseline and during the follow-up period. There were 94 subjects, with 180 minor recurrent aphthous ulcerations. The ulcers were distributed as 67, 57, and 56 ulcers for honey, topical corticosteroid, and Orabase treatment, respectively. There was a statistically significant difference between the honey group and the other two groups in terms of reduction of ulcer size, days of pain, and degree of erythema. No side effects were reported in any group. Honey was found to be effective and safe in reducing minor aphthous ulcer pain, size, and erythema in a Saudi cohort.


Looking Through a Social Lens: Conceptualising Social Aspects of Knowledge Management for Global Health Practitioners.
Knowledge management plays a critical role in global health. Global health practitioners require knowledge in every aspect of their jobs, and in resource-scarce contexts, practitioners must be able to rely on a knowledge management system to access the latest research and practice to ensure the highest quality of care. However, we suggest that there is a gap in the way knowledge management is primarily utilized in global health, namely, the systematic incorporation of human and social factors. In this paper, we briefly outline the evolution of knowledge management and then propose a conceptualization of knowledge management that incorporates human and social factors for use within a global health context. Our conceptualization of social knowledge management recognizes the importance of social capital, social learning, social software and platforms, and social networks, all within the context of a larger social system and driven by social benefit. We then outline the limitations and discuss future directions of our conceptualization, and suggest how this new conceptualization is essential for any global health practitioner in the business of managing knowledge.


Associations between diabetic retinopathy and plasma levels of high-sensitive C-reactive protein or von Willebrand factor in long-term type 1 diabetic patients.
To evaluate high-sensitive C-reactive protein (hs-CRP) and von Willebrand factor as possible plasma markers of diabetic retinopathy in a population-based cohort of type 1 diabetic patients. This was a cross-sectional study of 201 type 1 diabetic patients from a population-based cohort from Fyn County, Denmark. Plasma levels of hs-CRP and von Willebrand factor antigen were measured and related to the level of diabetic retinopathy (DR) as evaluated by dilated nine-field 45 degree monoscopic fundus photos captured by Topcon TRC-NWS6 and graded according to the Early Treatment Diabetic Retinopathy Study (ETDRS) adaptation of the modified Airlie House classification of DR. Median age and duration of diabetes were 58.7 and 43 years, respectively. Median levels (10th-90th percentile) of hs-CRP and von Willebrand factor antigen were 1.31 mg/l (0.37-13.3 mg/l) and 1.27 IU/ml (0.79-2.07 IU/ml), respectively. No or minimal DR (ETDRS-levels 10-20) was found in 16.4%, mild DR (ETDRS-level 35) in 19.4%, moderate DR (ETDRS-levels 43-47) in 11.0%, and 53.2% had proliferative diabetic retinopathy (PDR) corresponding to ETDRS-level 60 or more. In an age- and sex-adjusted model, patients in the highest quartile of hs-CRP were more likely to have PDR than patients in the lowest quartile (odds ratio: 2.59; 95% confidence interval: 1.09-6.12). However, this was no longer statistically significant in a multivariate model. Von Willebrand factor was not associated with PDR in any model. Even though patients with higher levels of hs-CRP were more likely to have PDR in an age- and sex-adjusted model, this was no longer statistically significant in a multivariate model. This indicates the importance of other risk factors like duration of diabetes, glycemic regulation, and smoking. We did not find any association between von Willebrand factor and diabetic retinopathy.


Asymmetric transmission of linearly polarized waves and polarization angle dependent wave rotation using a chiral metamaterial.
An electrically thin chiral metamaterial structure composed of four U-shaped split ring resonator pairs is utilized in order to realize polarization rotation that is dependent on the polarization of the incident wave at 6.2 GHz. The structure is optimized such that a plane wave that is linearly polarized at an arbitrary angle is an eigenwave of the system at this frequency. The analytical relation between the incident polarization and the polarization rotation is derived using transmission matrices. Furthermore, the proposed structure exhibits an asymmetric transmission of linearly polarized waves at 6.2 GHz. Plane waves traveling in opposite but perpendicular directions to the material plane are rotated by different angles. On the other hand, four incident polarization angles have been found for the same structure, at which the transmission is symmetric. The experiment results are in good agreement with the numerical results.


Ca2+ handling properties of microsomal subfractions of rat vas deferens smooth muscle.
The rat vas deferens smooth muscle microsomes on isopycnic centrifugation gave two fractions, namely F2 (15-30% sucrose) and F3 (30-40% sucrose), with comparable ATP-dependent azide-insensitive Ca2+-uptake capacities, although these fractions differed from each other in various enzyme marker activities. The fractions F2 and F3 also show similar pH profiles for the ATP-independent and ATP-dependent Ca2+ uptake, and similar ionized Ca2+-concentration dependence for the ATP-dependent Ca2+ uptake. However, the fractions F2 and F3 differ from each other in that: (a) F3 shows higher permeability to Ca2+, and (b) F3 shows higher stimulation of the ATP-dependent Ca2+ uptake by oxalate. The F3 fraction can also be used to obtain membrane vesicles loaded with Ca2+ oxalate in the presence of ATP. However, the yield of the Ca2+ oxalate enriched fraction is too low to permit their further characterization.


Intraspecific sequence variation and differential expression in starch synthase genes of Arabidopsis thaliana.
Natural accessions of Arabidopsis thaliana are a well-known system to measure levels of intraspecific genetic variation. Leaf starch content correlates negatively with biomass. Starch is synthesized by the coordinated action of many (iso)enzymes. Quantitatively dominant is the repetitive transfer of glucosyl residues to the non-reducing ends of α-glucans as mediated by starch synthases. In the genome of A. thaliana, there are five classes of starch synthases, designated as soluble starch synthases (SSI, SSII, SSIII, and SSIV) and granule-bound synthase (GBSS). Each class is represented by a single gene. The five genes are homologous in functional domains due to their common origin, but have evolved individual features as well. Here, we analyze the extent of genetic variation in these fundamental protein classes as well as possible functional implications on transcript and protein levels. Intraspecific sequence variation of the five starch synthases was determined by sequencing the entire loci including promoter regions from 30 worldwide distributed accessions of A. thaliana. In all genes, a considerable number of nucleotide polymorphisms was observed, both in non-coding and coding regions, and several amino acid substitutions were identified in functional domains. Furthermore, promoters possess numerous polymorphisms in potentially regulatory cis-acting regions. By realtime experiments performed with selected accessions, we demonstrate that DNA sequence divergence correlates with significant differences in transcript levels. Except for AtSSII, all starch synthase classes clustered into two or three groups of haplotypes, respectively. Significant difference in transcript levels among haplotype clusters in AtSSIV provides evidence for cis-regulation. By contrast, no such correlation was found for AtSSI, AtSSII, AtSSIII, and AtGBSS, suggesting trans-regulation. The expression data presented here point to a regulation by common trans-regulatory transcription factors which ensures a coordinated action of the products of these four genes during starch granule biosynthesis. The apparent cis-regulation of AtSSIV might be related to its role in the initiation of de novo biosynthesis of granules.


[The most frequent allergens in allergic contact dermatitis].
The prevalence of allergic contact dermatitis is high. Patch testing is the standard procedure to detect contact sensitivity. More than 3000 allergens are known, today. On the basis of the data of this study the top allergens are presented. The retrospective analysis is based on the patch test results collected by the Information Network of Departments of Dermatology in 2004. Data of 9948 patients (6175 female und 3773 male) referred for the evaluation of suspected contact allergy were analysed to determine the frequency of sensitization to the 10 most common contact allergens. The top ten allergens were nickel sulphate (frequency of sensitization standardized for sex and age 17.2 %), fragrance mix (7.2 %), balsam of Peru (6.7 %), cobalt chloride (6.5 %), potassium dichromate (5.3 %), colophony (4.6 %), lanolin alcohol (4.3 %), p-phenylenediamine (4.2 %), ammoniated mercury (3.5 %) und methyldibromoglutaronitrile/phenoxyethanol (3.4 %) in 2004. Even after healing of the skin lesions of allergic contact dermatitis, it must be considered that sensitization persists indefinitely. Therefore, patients have to be informed exactly about their relevant contact allergens to ensure complete avoidance. To do so, physicians need a detailed knowledge of the most common contact allergens which are discussed in this paper.


The discovery of a potent Nav1.3 inhibitor with good oral pharmacokinetics.
In this article, we describe the discovery of an aryl ether series of potent and selective Nav1.3 inhibitors. Based on structural analogy to a similar series of compounds we have previously shown bind to the domain IV voltage sensor region of Nav channels, we propose this series binds in the same location. We describe the development of this series from a published starting point, highlighting key selectivity and potency data, and several studies designed to validate Nav1.3 as a target for pain.


Using systematic reviews in clinical education.
Traditional educational methods change clinical practice only with considerable effort and difficulty. In particular, the teaching of critical appraisal in the setting of journal clubs does not increase the amount of medical research read by trainees. Experiential learning theory, corroborated by the success of problem-based learning, encourages us to link learning to the numerous medical questions that physicians generate while providing patient care. Systematic reviews can link these questions with the results of research that would otherwise be difficult to locate, read, and appraise. Systematic reviews are a uniquely powerful mechanism for teaching, and they offer teachers a new opportunity to model rational and effective use of information. Systematic reviews should be made available at clinical sites for use during "teachable moments." Resistance to the use of systematic reviews can be reduced by using existing journal clubs to teach about the strengths and limitations of these reviews. The point that systematic reviews are meant to assist, not replace, clinical decision making deserves emphasis in such teaching.


Gene Expression Profiling of Large Cell Lung Cancer Links Transcriptional Phenotypes to the New Histological WHO 2015 Classification.
Large cell lung cancer (LCLC) and large cell neuroendocrine carcinoma (LCNEC) constitute a small proportion of NSCLC. The WHO 2015 classification guidelines changed the definition of the debated histological subtype LCLC to be based on immunomarkers for adenocarcinoma and squamous cancer. We sought to determine whether these new guidelines also translate into the transcriptional landscape of lung cancer, and LCLC specifically. Gene expression profiling was performed by using Illumina V4 HT12 microarrays (Illumina, San Diego, CA) on samples from 159 cases (comprising all histological subtypes, including 10 classified as LCLC WHO 2015 and 14 classified as LCNEC according to the WHO 2015 guidelines), with complimentary mutational and immunohistochemical data. Derived transcriptional phenotypes were validated in 199 independent tumors, including six WHO 2015 LCLCs and five LCNECs. Unsupervised analysis of gene expression data identified a phenotype comprising 90% of WHO 2015 LCLC tumors, with characteristics of poorly differentiated proliferative cancer, a 90% tumor protein p53 gene (TP53) mutation rate, and lack of well-known NSCLC oncogene driver alterations. Validation in independent data confirmed aggregation of WHO 2015 LCLCs in the specific phenotype. For LCNEC tumors, the unsupervised gene expression analysis suggested two different transcriptional patterns corresponding to a proposed genetic division of LCNEC tumors into SCLC-like and NSCLC-like cancer on the basis of TP53 and retinoblastoma 1 gene (RB1) alteration patterns. Refined classification of LCLC has implications for diagnosis, prognostics, and therapy decisions. Our molecular analyses support the WHO 2015 classification of LCLC and LCNEC tumors, which herein follow different tumorigenic paths and can accordingly be stratified into different transcriptional subgroups, thus linking diagnostic immunohistochemical staining-driven classification with the transcriptional landscape of lung cancer.


Fullerene and corannulene derivatives acting as insulators of Cl(-) and BeH2.
The capacity of corannulene and its benzo-derivatives CxH10 (x = 20-60) as prototypes of non-planar π-aromatic systems which mimic the end of carbon nanotubes to act as insulators between BeH2 and Cl(-) chemical entities has been explored by means of M06-2x/cc-pVDZ and M06-2x/6-311+G(d) calculations. For the sake of completeness, the set investigated includes also fullerene, C60. All these aromatic derivatives lead to stable binary complexes either with BeH2 or halogen (Cl(-)) anions. For BeH2, however, only the complexes in which the interaction involves the convex face of the aromatic system are stable. No significant changes are observed when the binding energies (BEs) of the BeH2 complexes are compared to those involving planar aromatic compounds, but the ones involving Cl(-) with the concave face of the aromatic moiety can be very large, because its curvature favors many contacts of the anion with the carbon atoms of the π-aromatic system. The formation of these binary complexes changes to a large extent the electrostatic potential on the free face of the aromatic system leading to a mutual reinforcement of both interactions, the beryllium bond and the interaction with Cl(-), when the ternary complexes are formed. As a result, the BEs for the triads are larger than the sum of the BEs of the corresponding binary complexes and the distances between the aromatic subunit and BeH2 or Cl(-) become shorter in the triads than in the binary complexes. A MBIE analysis also indicates that the enhanced stability of ternary complexes arises mainly from the reinforcement of the beryllium bonds as well as from the three-body terms. An exploration of all the minima for BeH2:C60H10:Cl(-) shows that BeH2 binds preferentially to the peripheral aromatic rings than those in the more curved region.


Sports helmets now and in the future.
The paper reports on a symposium on sports helmets and presents a synthesis of information and opinion from a range of presenters and disciplines. A review of the literature shows that helmets play an important role in head injury prevention and control. Helmets have been shown to be very efficacious and effective in a range of sports and in preventing specific head injury risks, especially moderate to severe head injury. The symposium emphasised the importance of helmet standards and the need for further development. There are calls for helmets that address the needs of competitive (elite) athletes separate to helmets for recreational athletes. Deficiencies in the evidence base for head injury risks and helmet efficacy and effectiveness were identified in some sports. Issues in designing helmets that are suitable to prevent severe head injuries and concussion were discussed and explained from biomechanical and engineering perspectives. The need to evaluate helmet performance in oblique impacts and incorporate this into standards was covered in a number of presentations. There are emerging opportunities with in-helmet technology to improve impact performance or to measure impact exposure. In-helmet technology as it matures may provide critical information on the severity of the impact, the location of the injured athlete, for example, snowboarder, and assist in the retrieval and immediate, as well as the long-term medical management of the athlete. It was identified that athletes, families and sports organisations can benefit from access to information on helmet performance. The importance of selecting the appropriate-sized helmet and ensuring that the helmet and visor were adjusted and restrained optimally was emphasised. The translation pathway from the science to new and better helmets is the development of appropriate helmet standards and the requirement for only helmets to be used that are certified to those standards.


Force distribution is more important than its intensity!
A common question about root resorption is raised in orthodontic practice: What is more important, the intensity of force or its distribution along the root, periodontal and alveolar structures? Diffuse distribution of forces applied to periodontal tissues during tooth movement tends not to promote neither extensive areas of cell matrix hyalinization nor significant death of cementoblasts that lead to root resorption. However, focal distribution or concentration of forces within a restricted area--as it occurs in tipping movements, even with forces of lower intensity--tend to induce extensive areas of hyalinization and focal death of cementoblasts, which is commonly associated with root resorption. In tipping movements, the apical regions tend to concentrate more forces, in addition to wounding the cementoblasts due to the smaller dimension of their root structure as well as their cone shape. For this reason, there is an increase in root resorption. In the cervical region, on the other hand, the large area resulting from a large diameter and bone crown deflection tends to reduce the effects of forces, even when they are more concentrated, thus rarely inducing death of cementoblasts and root resorption.


Age and oestrus cycle-related changes in glucocorticoid excretion and wheel-running activity in female mice carrying mutations in the circadian clock genes Per1 and Per2.
In mammals, numerous physiological and behavioural functions are controlled by an endogenous circadian clock located in the suprachiasmatic nuclei (SCN). Within the SCN neurons, clock genes such as Per1 and Per2 interact in a molecular clockwork regulating the expression of hundreds of output genes. Through the timed release of humoral and neuronal signals, the rhythmicity of numerous biological processes, including reproductive behaviour, the oestrus cycle and endocrine parameters is controlled by the SCN. Mutations in Per genes in mice affect a wide array of physiological functions. Interestingly, most of these studies use only male animals, thus neglecting potential gender-specificities in clock function. In an attempt to broaden this perspective we have investigated the impact of Per1 and Per2 mutations on both glucocorticoid (GC) metabolite excretion and locomotor activity in relation to age and oestrus cycle stage of female mice. We show that the Per2 mutation dampens daily GC rhythms in young adult females. While locomotor activity does not vary along the different oestrus stages in Per2 mutant females, oestrus effects on GC excretion and locomotor activity are largely comparable between Per1 mutants and wild-type animals. 20 month-old, acyclic Per1 and wild-type females show reduced GC levels when compared to young adults while aged Per2 mutants retain their normal GC rhythmicity. Correlating with this, onsets of locomotor activity do not change with age in Per2 mutant females. Together, our data highlight specific roles for Per1 and Per2 in both the regulation of locomotor activity and endocrine functions in the female organism.


A combination test for detection of gene-environment interaction in cohort studies.
Identifying gene-environment (G-E) interactions can contribute to a better understanding of disease etiology, which may help researchers develop disease prevention strategies and interventions. One big criticism of studying G-E interaction is the lack of power due to sample size. Studies often restrict the interaction search to the top few hundred hits from a genome-wide association study or focus on potential candidate genes. In this paper, we test interactions between a candidate gene and an environmental factor to improve power by analyzing multiple variants within a gene. We extend recently developed score statistic based genetic association testing approaches to the G-E interaction testing problem. We also propose tests for interaction using gene-based summary measures that pool variants together. Although it has recently been shown that these summary measures can be biased and may lead to inflated type I error, we show that under several realistic scenarios, we can still provide valid tests of interaction. These tests use significantly less degrees of freedom and thus can have much higher power to detect interaction. Additionally, we demonstrate that the iSeq-aSum-min test, which combines a gene-based summary measure test, iSeq-aSum-G, and an interaction-based summary measure test, iSeq-aSum-I, provides a powerful alternative to test G-E interaction. We demonstrate the performance of these approaches using simulation studies and illustrate their performance to study interaction between the SNPs in several candidate genes and family climate environment on alcohol consumption using the Minnesota Center for Twin and Family Research dataset.


Mathematical model for low density lipoprotein (LDL) endocytosis by hepatocytes.
Individuals with elevated levels of plasma low density lipoprotein (LDL) cholesterol (LDL-C) are considered to be at risk of developing coronary heart disease. LDL particles are removed from the blood by a process known as receptor-mediated endocytosis, which occurs mainly in the liver. A series of classical experiments delineated the major steps in the endocytotic process; apolipoprotein B-100 present on LDL particles binds to a specific receptor (LDL receptor, LDL-R) in specialized areas of the cell surface called clathrin-coated pits. The pit comprising the LDL-LDL-R complex is internalized forming a cytoplasmic endosome. Fusion of the endosome with a lysosome leads to degradation of the LDL into its constituent parts (that is, cholesterol, fatty acids, and amino acids), which are released for reuse by the cell, or are excreted. In this paper, we formulate a mathematical model of LDL endocytosis, consisting of a system of ordinary differential equations. We validate our model against existing in vitro experimental data, and we use it to explore differences in system behavior when a single bolus of extracellular LDL is supplied to cells, compared to when a continuous supply of LDL particles is available. Whereas the former situation is common to in vitro experimental systems, the latter better reflects the in vivo situation. We use asymptotic analysis and numerical simulations to study the longtime behavior of model solutions. The implications of model-derived insights for experimental design are discussed.


Highly recurrent TERT promoter mutations in human melanoma.
Systematic sequencing of human cancer genomes has identified many recurrent mutations in the protein-coding regions of genes but rarely in gene regulatory regions. Here, we describe two independent mutations within the core promoter of telomerase reverse transcriptase (TERT), the gene coding for the catalytic subunit of telomerase, which collectively occur in 50 of 70 (71%) melanomas examined. These mutations generate de novo consensus binding motifs for E-twenty-six (ETS) transcription factors, and in reporter assays, the mutations increased transcriptional activity from the TERT promoter by two- to fourfold. Examination of 150 cancer cell lines derived from diverse tumor types revealed the same mutations in 24 cases (16%), with preliminary evidence of elevated frequency in bladder and hepatocellular cancer cells. Thus, somatic mutations in regulatory regions of the genome may represent an important tumorigenic mechanism.


Delayed symptom onset and increased life expectancy in Sandhoff disease mice treated with N-butyldeoxynojirimycin.
Sandhoff disease is a neurodegenerative disorder resulting from the autosomal recessive inheritance of mutations in the HEXB gene, which encodes the beta-subunit of beta-hexosaminidase. GM2 ganglioside fails to be degraded and accumulates within lysosomes in cells of the periphery and the central nervous system (CNS). There are currently no therapies for the glycosphingolipid lysosomal storage diseases that involve CNS pathology, including the GM2 gangliosidoses. One strategy for treating this and related diseases is substrate deprivation. This would utilize an inhibitor of glycosphingolipid biosynthesis to balance synthesis with the impaired rate of catabolism, thus preventing storage. One such inhibitor is N-butyldeoxynojirimycin, which currently is in clinical trials for the potential treatment of type 1 Gaucher disease, a related disease that involves glycosphingolipid storage in peripheral tissues, but not in the CNS. In this study, we have evaluated whether this drug also could be applied to the treatment of diseases with CNS storage and pathology. We therefore have treated a mouse model of Sandhoff disease with the inhibitor N-butyldeoxynojirimycin. The treated mice have delayed symptom onset, reduced storage in the brain and peripheral tissues, and increased life expectancy. Substrate deprivation therefore offers a potentially general therapy for this family of lysosomal storage diseases, including those with CNS disease.


Latitude, birth date, and allergy.
The space and time distribution of risk factors for allergic diseases may provide insights into disease mechanisms. Allergy is believed to vary by month of birth, but multinational studies taking into account latitude have not been conducted. A questionnaire was distributed in 54 centres to a representative sample of 20- to 44-y-old men and women mainly in Europe but also including regions in North Africa, India, North America, Australia, and New Zealand. Data from 200,682 participants were analyzed. The median prevalence of allergic rhinitis was 22%, with a substantial variation across centres. Overall, allergic rhinitis decreased with geographical latitude, but there were many exceptions. No increase in prevalence during certain winters could be observed. Also, no altered risk by birth month was found, except borderline reduced risks in September and October. Effect estimates obtained by a multivariate analysis of total and specific IgE values in 18,085 individuals also excluded major birth month effects and confirmed the independent effect of language grouping. Neither time point of first exposure to certain allergens nor early infections during winter months seems to be a major factor for adult allergy. Although there might be effects of climate or environmental UV exposure by latitude, influences within language groups seem to be more important, reflecting so far unknown genetic or cultural risk factors.


Analysis of high-depth sequence data for studying viral diversity: a comparison of next generation sequencing platforms using Segminator II.
Next generation sequencing provides detailed insight into the variation present within viral populations, introducing the possibility of treatment strategies that are both reactive and predictive. Current software tools, however, need to be scaled up to accommodate for high-depth viral data sets, which are often temporally or spatially linked. In addition, due to the development of novel sequencing platforms and chemistries, each with implicit strengths and weaknesses, it will be helpful for researchers to be able to routinely compare and combine data sets from different platforms/chemistries. In particular, error associated with a specific sequencing process must be quantified so that true biological variation may be identified. Segminator II was developed to allow for the efficient comparison of data sets derived from different sources. We demonstrate its usage by comparing large data sets from 12 influenza H1N1 samples sequenced on both the 454 Life Sciences and Illumina platforms, permitting quantification of platform error. For mismatches median error rates at 0.10 and 0.12%, respectively, suggested that both platforms performed similarly. For insertions and deletions median error rates within the 454 data (at 0.3 and 0.2%, respectively) were significantly higher than those within the Illumina data (0.004 and 0.006%, respectively). In agreement with previous observations these higher rates were strongly associated with homopolymeric stretches on the 454 platform. Outside of such regions both platforms had similar indel error profiles. Additionally, we apply our software to the identification of low frequency variants. We have demonstrated, using Segminator II, that it is possible to distinguish platform specific error from biological variation using data derived from two different platforms. We have used this approach to quantify the amount of error present within the 454 and Illumina platforms in relation to genomic location as well as location on the read. Given that next generation data is increasingly important in the analysis of drug-resistance and vaccine trials, this software will be useful to the pathogen research community. A zip file containing the source code and jar file is freely available for download from http://www.bioinf.manchester.ac.uk/segminator/.


Functional coherence of striatal resting-state networks is modulated by striatal iron content.
Resting-state spontaneous fluctuations have revealed individual differences in the functional architecture of brain networks. Previous research indicates that the striatal network shows alterations in neurological conditions but also in normal aging. However, the neurobiological mechanisms underlying individual differences in striatal resting-state networks (RSNs) have been less explored. One candidate that may account for individual differences in striatal spontaneous activity is the level of local iron accumulation. Excessive iron in the striatum has been linked to a loss of structural integrity and reduced brain activity during task performance in aging. Using independent component analysis in a sample of 42 younger and older adults, we examined whether higher striatal iron content, quantified using relaxometry, underlies individual differences in spontaneous fluctuations of RSNs in general, and of the striatum in particular. Higher striatal iron content was linked to lower spontaneous coherence within both caudate and putamen RSNs regardless of age. No such links were observed for other RSNs. Moreover, the number of connections between the putamen and other RSNs was negatively associated with iron content, suggesting that iron modulated the degree of cross-talk between the striatum and cerebral cortex. Importantly, these associations were primarily driven by the older group. Finally, a positive association was found between coherence in the putamen and motor performance, suggesting that this spontaneous activity is behaviorally meaningful. A follow-up mediation analysis also indicated that functional connectivity may mediate the link between striatal iron and motor performance. Our preliminary findings suggest that striatal iron potentially accounts for individual differences in spontaneous striatal fluctuations, and might be used as a locus of intervention.


Comparison of four Fitbit and Jawbone activity monitors with a research-grade ActiGraph accelerometer for estimating physical activity and energy expenditure.
Consumer-based physical activity (PA) monitors have become popular tools to track PA behaviours. Currently, little is known about the validity of the measurements provided by consumer monitors. We aimed to compare measures of steps, energy expenditure (EE) and active minutes of four consumer monitors with one research-grade accelerometer within a semistructured protocol. Thirty men and women (18-80 years old) wore Fitbit One (worn at the waist), Fitbit Zip (waist), Fitbit Flex (wrist), Jawbone UP24 (wrist) and one waist-worn research-grade accelerometer (ActiGraph) while participating in an 80 min protocol. A validated EE prediction equation and active minute cut-points were applied to ActiGraph data. Criterion measures were assessed using direct observation (step count) and portable metabolic analyser (EE, active minutes). A repeated measures analysis of variance (ANOVA) was used to compare differences between consumer monitors, ActiGraph, and criterion measures. Similarly, a repeated measures ANOVA was applied to a subgroup of subjects who didn't cycle. Participants took 3321±571 steps, had 28±6 active min and expended 294±56 kcal based on criterion measures. Comparatively, all monitors underestimated steps and EE by 13%-32% (p<0.01); additionally the Fitbit Flex, UP24, and ActiGraph underestimated active minutes by 35%-65% (p<0.05). Underestimations of PA and EE variables were found to be similar in the subgroup analysis. Consumer monitors had similar accuracy for PA assessment as the ActiGraph, which suggests that consumer monitors may serve to track personal PA behaviours and EE. However, due to discrepancies among monitors, individuals should be cautious when comparing relative and absolute differences in PA values obtained using different monitors.


Hypoxia selectively inhibits KCNA5 channels in pulmonary artery smooth muscle cells.
Acute hypoxia induces pulmonary vasoconstriction and chronic hypoxia causes pulmonary vascular remodeling characterized by significant vascular medial hypertrophy. Electromechanical and pharmacomechanical mechanisms are involved in regulating pulmonary vasomotor tone, while changes in cytosolic Ca2+ concentration ([Ca2+](cyt)) are an important signal in regulating contraction and proliferation of pulmonary artery smooth muscle cells (PASMC). Hypoxia-induced increases in [Ca2+](cyt) are, in part, mediated by selective inhibition of voltage-gated K+ (Kv) channels in PASMC. Kv1.5, encoded by the KCNA5 gene, is a Kv channel alpha subunit that forms functional homotetrameric and heterotetrameric Kv channels in PASMC. Activity of Kv channels contributes to the regulation of resting membrane potential. Overexpression of the human KCNA5 gene in rat PASMC and other cell types increases whole-cell Kv currents and causes membrane hyperpolarization. However, acute hypoxia only reduced Kv currents in KCNA5-transfected PASMC. These results provide compelling evidence that Kv1.5 is an important hypoxia-sensitive Kv channel in PASMC, contributing to regulation of membrane potential and intracellular Ca2+ homeostasis during hypoxia. This hypoxia-sensitive mechanism essential for inhibiting Kv1.5 channel activity is exclusively present in PASMC.


Pilot study using molecular profiling of patients' tumors to find potential targets and select treatments for their refractory cancers.
To compare the progression-free survival (PFS) using a treatment regimen selected by molecular profiling (MP) of a patient's tumor with the PFS for the most recent regimen on which the patient had experienced progression (ie, patient as his own control). Patients with refractory metastatic cancer had tissue samples submitted for MP in two formats including formalin-fixed tissue for immunohistochemistry and fluorescent in situ hybridization assays and immediately frozen tissue for oligonucleotide microarray (MA) gene expression assays (all performed in a Clinical Laboratory Improvement Amendments [CLIA]-certified laboratory). The MP approach was deemed of clinical benefit for the individual patient who had a PFS ratio (PFS on MP-selected therapy/PFS on prior therapy) of ≥ 1.3. In 86 patients who had MP attempted, there was a molecular target detected in 84 (98%). Sixty-six of the 84 patients were treated according to MP results. Eighteen (27%) of 66 patients had a PFS ratio of ≥ 1.3 (95% CI, 17% to 38%; one-sided, one-sample P = .007). Therefore, the null hypothesis (that ≤ 15% of this patient population would have a PFS ratio of ≥ 1.3) was rejected. It is possible to identify molecular targets in patients' tumors from nine different centers across the United States. In 27% of patients, the MP approach resulted in a longer PFS on an MP-suggested regimen than on the regimen on which the patient had just experienced progression. Issues to be considered in interpretation of this study include limited prior experience with patients as their own controls as a study end point and overall patient attrition.


Regular peanut consumption improves plasma lipid levels in healthy Ghanaians.
To determine whether the daily intake of 2,092 kJ (500 kcal) from peanuts will improve the lipid profiles and diet quality of healthy Ghanaians. A 30-week, randomized, cross-over trial study was conducted with healthy adults. There were three treatment arms: Treatment 1 (T1), subjects were provided 2,092 kJ/day (500 kcal/day) peanuts to incorporate into their daily diet for 8 weeks at any time and in any form they chose; Treatment 2 (T2), subjects were provided 2,092 kJ/day (500 kcal/day) peanuts and were instructed to consume them in addition to their customary daily diet for 3 weeks; Treatment 3 (T3), substitution of 2,092 kJ/day fat, with energy from peanuts. Total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol and triglyceride were measured at baseline, week 4 and week 8 (T1 and T3) or at baseline and week 3 (T2). Three-day dietary intake records were kept during each treatment. There was significant decrease in total cholesterol (7.2%) and triacylglycerol (20.0%) after T1. However, individually, high-density lipoprotein-cholesterol and low-density lipoprotein-cholesterol levels did not change significantly. Total fat intake increased by 9%, due to elevations of monounsaturated fatty acid of 60% and polyunsaturated fatty acid of 50%. Similar non-significant trends were observed during T2 and T3. The results suggest that regular consumption of peanuts lowers the total cholesterol and triacylglycerol concentrations among healthy Ghanaians. Regular consumption of peanuts should therefore be encouraged.


A Model for Metal Adsorption on Montmorillonite.
A consistent thermodynamic model is developed for metal sorption on expanding 2:1 layer clays such as montmorillonite. The particle of clay, including lamellae and interlayers, is represented as a porous solid bearing a permanent negative charge (resulting from isomorphic substitution) with an infinite plane interface (i.e., edges) with the solution. Cation exchange occurs inside the clay particle as the result of the negative potential of the clay. Surface complexation reactions take place at the interface whose surface charge and potential are pH dependent. The potential in the bulk of the clay and near the interface, as well as the surface potential-surface charge density relation, are calculated taking into account the effect of the permanent negative charge. The results are discussed and compared with the classic Gouy-Chapman theory. A subroutine (Clayeql) with the new potential-charge relationships is implemented in the thermodynamic equilibrium program Mineql +3.0 and is used to fit an extensive published experimental data set on adsorption of transition metals on montmorillonite. The model is shown not only to fit satisfactorily all the data, but also to explain specific features of adsorption on clays compared to oxides. In particular, the increase in the surface concentration of protons with decreasing ionic strength is successfully reproduced and the weaker dependence of metal sorption on pH compared to oxides is correctly fitted. Copyright 1999 Academic Press.


Natural history of progressive ischemic stroke in a population treated with heparin.
Data on the acute natural history of progressive stroke with or without heparin treatment are limited. To define the acute course of patients treated with heparin for progressive stroke, we examined the charts of 69 such patients identified through the Cornell Neurology Database from October 1979 to June 1985. Analysis included determining whether further clinical deterioration or hemorrhagic complications were associated with readily identifiable clinical or laboratory variables. Twenty-five patients (36%) continued to deteriorate while receiving heparin, another two (3%) worsened due to intracerebral hemorrhage, and a total of 10 patients (14%) had bleeding complications. No clinical features or heparin dosing regimens distinguished the patients likely to benefit from heparin. Clinical progression or hemorrhage did not correlate with the level of anticoagulation as measured by the average heparin dose per day or the mean partial thromboplastin time. Without unequivocal evidence demonstrating heparin's ineffectiveness for progressive stroke, many clinicians managing such patients will continue to use heparin. Our results suggest that this decision should not be governed by such clinical features as a patient's age or sex or by the vascular distribution of the stroke. Furthermore, frequent measurement of and overzealous efforts to adjust the partial thromboplastin time may be unnecessary since it does not correlate with outcome.


IL-18BP is a secreted immune checkpoint and barrier to IL-18 immunotherapy.
Cytokines were the first modern immunotherapies to produce durable responses in patients with advanced cancer, but they have only modest efficacy and limited tolerability1,2. In an effort to identify alternative cytokine pathways for immunotherapy, we found that components of the interleukin-18 (IL-18) pathway are upregulated on tumour-infiltrating lymphocytes, suggesting that IL-18 therapy could enhance anti-tumour immunity. However, recombinant IL-18 previously did not demonstrate efficacy in clinical trials3. Here we show that IL-18BP, a high-affinity IL-18 decoy receptor, is frequently upregulated in diverse human and mouse tumours and limits the anti-tumour activity of IL-18 in mice. Using directed evolution, we engineered a 'decoy-resistant' IL-18 (DR-18) that maintains signalling potential but is impervious to inhibition by IL-18BP. Unlike wild-type IL-18, DR-18 exerted potent anti-tumour effects in mouse tumour models by promoting the development of poly-functional effector CD8+ T cells, decreasing the prevalence of exhausted CD8+ T cells that express the transcriptional regulator of exhaustion TOX, and expanding the pool of stem-like TCF1+ precursor CD8+ T cells. DR-18 also enhanced the activity and maturation of natural killer cells to effectively treat anti-PD-1 resistant tumours that have lost surface expression of major histocompatibility complex class I molecules. These results highlight the potential of the IL-18 pathway for immunotherapeutic intervention and implicate IL-18BP as a major therapeutic barrier.


Donor arterial variations in liver transplantation: management and outcome of 527 consecutive grafts.
The impact of donor arterial variations and their management was investigated retrospectively in 527 consecutive allografts. Anomalous arteries were found in 161 grafts (30.6 per cent). There was no significant difference in the overall incidence of arterial complications between grafts with normal (3.6 per cent) and abnormal (5.6 per cent) anatomy. However, there was a higher incidence of arterial complications in transplants requiring multiple arterial anastomoses (P = 0.02), or anastomosis of donor vessels to recipient aorta with (P = 0.0003) or without (P = 0.04) an interposition graft for arterial reconstruction. The incidence of biliary complications was similar in grafts with a normal (18.8 per cent) or anomalous (18.0 per cent) arterial supply. Anomalies of hepatic arterial anatomy occur in one-third of all livers and do not compromise graft outcome unless multiple anastomoses or direct anastomosis to the recipient aorta are required for arterial reconstruction.


Peptidoglycan derived from Staphylococcus epidermidis induces Connexin43 hemichannel activity with consequences on the innate immune response in endothelial cells.
Gram-positive bacterial cell wall components including PGN (peptidoglycan) elicit a potent pro-inflammatory response in diverse cell types, including endothelial cells, by activating TLR2 (Toll-like receptor 2) signalling. The functional integrity of the endothelium is under the influence of a network of gap junction intercellular communication channels composed of Cxs (connexins) that also form hemichannels, signalling conduits that are implicated in ATP release and purinergic signalling. PGN modulates Cx expression in a variety of cell types, yet effects in endothelial cells remain unresolved. Using the endothelial cell line b.End5, a 6 h challenge with PGN induced IL-6 (interleukin 6), TLR2 and Cx43 mRNA expression that was associated with enhanced Cx43 protein expression and gap junction coupling. Cx43 hemichannel activity, measured by ATP release from the cells, was induced following 15 min of exposure to PGN. Inhibition of hemichannel activity with carbenoxolone or apyrase prevented induction of IL-6 and TLR2 mRNA expression by PGN, but had no effect on Cx43 mRNA expression levels. In contrast, knockdown of TLR2 expression had no effect on PGN-induced hemichannel activity, but reduced the level of TLR2 and Cx43 mRNA expression following 6 h of PGN challenge. PGN also acutely induced hemichannel activity in HeLa cells transfected to express Cx43, but had no effect in Cx43-deficient HeLa OHIO cells. All ATP responses were blocked with Cx-specific channel blockers. We conclude that acute Cx43 hemichannel signalling plays a role in the initiation of early innate immune responses in the endothelium.


Interleukin-6 modulates graft-versus-host responses after experimental allogeneic bone marrow transplantation.
The graft-versus-tumor (GVT) effect is a potent form of immunotherapy against many hematologic malignancies and some solid tumors. The beneficial GVT effect after allogeneic bone marrow transplantation (BMT) is tightly linked to its most significant complication, graft-versus-host disease (GVHD). The role of interleukin-6 (IL-6) after allogeneic BMT is not well understood. This study used a series of complementary knockout and antibody blockade strategies to analyze the impact of IL-6 in multiple clinically relevant murine models of GVHD and GVT. We examined the effect of the source of IL-6 by analyzing the role IL-6 deficiency in donor T cells, donor bone marrow or in host tissues. We confirmed and extended the relevance of IL-6 deficiency on GVHD and GVT by treating BMT recipients with anti-mouse IL-6 receptor (IL-6R), MR16-1. Deficiency of IL-6 in donor T cells led to prolongation of survival. Total inhibition of IL-6 with MR16-1 caused an even greater reduction in GVHD-induced mortality. The reduction in GVHD was independent of the direct effects on T effector cell expansion or donor regulatory T cells. GVT responses were preserved after treatment with MR16-1. MR16-1 treatment reduced GVHD and preserved sufficient GVT. Tocilizumab, a humanized anti-IL-6R monoclonal antibody (mAb), is approved in several countries including the United States and European Union for the treatment of rheumatoid arthritis and other inflammatory diseases. Blockade of IL-6 with anti-IL-6R mAb therapy may be testable in clinical trials as an adjunct to prevent GVHD in BMT patients without a significant loss of GVT.


Two-year clinical outcomes of radiofrequency focal ablation using a navigable plasma disc decompression device in patients with lumbar disc herniation: efficacy and complications.
To report on the 2-year clinical outcomes of focal ablation using a navigable plasma disc decompression device in patients with lumbar herniated nucleus pulposus (HNP). This was a prospective, single-cohort study conducted in a clinical center. A total of 170 patients with lumbar HNP were assessed for pain intensity using the visual analog scale (VAS), for disability level using the Oswestry Disability Questionnaire (ODI), for health-related quality of life using the short form-36 version 2 of the bodily pain scale (SF-36 BP), and for the angles of passive straight leg raise (SLR) test. The herniated portions of the target discs were ablated using a navigable catheter under a well-instructed protocol with informed consent. Outcome data were prospectively collected before the procedure: 1 week after the procedure: and then 1, 3, 6, 12, and 24 months postoperatively. For statistical analysis, repeated-measures analysis of variance was performed. Two years after the procedure, the mean VAS decreased from 7.1±1.7 to 2.1±1.9, the mean ODI decreased from 50.9±17.2 to 20.3% ± 14.6%, and the mean SF-36 BP increased from 38.8±8.4 to 45.8±9.4 (P<0.05). On the SLR test, the angular change after 2 years improved considerably from 51.2±17.3 to 85.0±9.3 degrees. There was 90%-100% VAS improvement in 25.9% and 50%-90% VAS improvement in 52.4% of the patients; 1.8% experienced pain aggravation compared with the initial VAS. Two subjects showed short-term foot drop, whereas one subject showed the severe complication of foot drop for more than 6 months. The recurrence rate ranged from 4.7% to 11.5%. These findings suggest that focal ablation of herniated discs using a navigable disc decompression device is worth considering for relieving pain related to lumbar HNP. However, the development of more advanced technologies and methods for safety and efficiency is necessary.


Active and passive intranasal immunizations with streptococcal surface protein C5a peptidase prevent infection of murine nasal mucosa-associated lymphoid tissue, a functional homologue of human tonsils.
C5a peptidase, also called SCPA (surface-bound C5a peptidase), is a surface-bound protein on group A streptococci (GAS), etiologic agents for a variety of human diseases including pharyngitis, impetigo, toxic shock, and necrotizing fasciitis, as well as the postinfection sequelae rheumatic fever and rheumatic heart disease. This protein is highly conserved among different serotypes and is also expressed in human isolates of group B, C, and G streptococci. Human tonsils are the primary reservoirs for GAS, maintaining endemic disease across the globe. We recently reported that GAS preferentially target nasal mucosa-associated lymphoid tissue (NALT) in mice, a tissue functionally analogous to human tonsils. Experiments using a C5a peptidase loss-of-function mutant and an intranasal infection model showed that this protease is required for efficient colonization of NALT. An effective vaccine should prevent infection of this secondary lymphoid tissue; therefore, the potential of anti-SCPA antibodies to protect against streptococcal infection of NALT was investigated. Experiments showed that GAS colonization of NALT was significantly reduced following intranasal immunization of mice with recombinant SCPA protein administered alone or with cholera toxin, whereas a high degree of GAS colonization of NALT was observed in control mice immunized with phosphate-buffered saline only. Moreover, administration of anti-SCPA serum by the intranasal route protected mice against streptococcal infection. These results suggest that intranasal immunization with SCPA would prevent colonization and infection of human tonsils, thereby eliminating potential reservoirs that maintain endemic disease.


Biogeography of the two major arbovirus mosquito vectors, Aedes aegypti and Aedes albopictus (Diptera, Culicidae), in Madagascar.
In the past ten years, the Indian Ocean region has been the theatre of severe epidemics of chikungunya and dengue. These outbreaks coincided with a high increase in populations of Aedes albopictus that outcompete its sister taxon Aedes aegypti in most islands sampled. The objective of this work was to update the entomological survey of the two Aedes species in the island of Madagascar which has to face these arboviroses. The sampling of Aedes mosquitoes was conducted during two years, from October 2007 to October 2009, in fifteen localities from eight regions of contrasting climates. Captured adults were identified immediately whereas immature stages were bred until adult stage for determination. Phylogenetic analysis was performed using two mtDNA genes, COI and ND5 and trees were constructed by the maximum likelihood (ML) method with the gene time reversible (GTR) model. Experimental infections with the chikungunya virus strain 06.21 at a titer of 107.5 pfu/mL were performed to evaluate the vector competence of field-collected mosquitoes. Disseminated infection rates were measured fourteen days after infection by immunofluorescence assay performed on head squashes. The species Aedes aegypti was detected in only six sites in native forests and natural reserves. In contrast, the species Aedes albopictus was found in 13 out of the 15 sites sampled. Breeding sites were mostly found in man-made environments such as discarded containers, used tires, abandoned buckets, coconuts, and bamboo cuts. Linear regression models showed that the abundance of Ae. albopictus was significantly influenced by the sampling region (F = 62.00, p < 2.2 × 10(-16)) and period (F = 36.22, p = 2.548 × 10(-13)), that are associated with ecological and climate variations. Phylogenetic analysis of the invasive Ae. albopictus distinguished haplotypes from South Asia and South America from those of Madagascar, but the markers used were not discriminant enough to discern Malagasy populations. The experimental oral infection method showed that six Ae. albopictus populations exhibited high dissemination infection rates for chikungunya virus ranging from 98 to 100%. In Madagascar, Ae. albopictus has extended its geographical distribution whereas, Ae. aegypti has become rare, contrasting with what was previously observed. Changes are predominantly driven by human activities and the rainfall regime that provide suitable breeding sites for the highly anthropophilic mosquito Ae. albopictus. Moreover, these populations were found to be highly susceptible to chikungunya virus. In the light of this study, Ae. albopictus may have been involved in the recent outbreaks of chikungunya and dengue epidemics in Madagascar, and consequently, control measures should be promoted to limit its current expansion.


Differential expression of HIF-1α in CD44+CD24-/low breast ductal carcinomas.
Cancer stem cell (CSC) hypothesis postulates that tumors are maintained by a self-renewing CSC population that is also capable of differentiating into non-self-renewing cell populations that constitute the bulk of tumor. Stem cells renewal and differentiation can be directly influenced by the oxygen levels of determined tissues, probably by the reduction of oxidative DNA damage in hypoxic regions, thus leading to a friendlier microenvironment, regarding to clonal expansion and for resistance to chemotherapeutic regimens. Furthermore, there have been strong data indicating a pivotal role of hypoxic niche in cancer stem cells development. There are evidence that hypoxia could drive the maintenance of CSC, via HIF-1α expression, but it still to be determined whether hypoxia markers are expressed in breast tumors presenting CD44+CD24-/low immunophenotype. Immunohistochemical analysis of CD44+CD24-/low expression and its relationship with hypoxia markers and clinical outcome were evaluated in 253 samples of breast ductal carcinomas. Double-immunolabeling was performed using EnVision Doublestain System (Dako, Carpinteria, CA, USA). Slides were then scanned into high-resolution images using Aperio ScanScope XT and then, visualized in the software Image Scope (Aperio, Vista, CA, USA). In univariate analysis, CD44+CD24-/low expression showed association with death due to breast cancer (p = 0.035). Breast tumors expressing CD44+CD24-/low immunophenotype showed relationship with HIF-1α (p = 0.039) and negativity for HER-2 (p = 0.013). Considering that there are strong evidences that the fraction of a tumour considered to be cancer stem cells is plastic depending upon microenvironmental signals, our findings provide further evidence that hypoxia might be related to the worse prognosis found in CD44+CD24-/low positive breast tumors.


Cdc42 and the actin-related protein/neural Wiskott-Aldrich syndrome protein network mediate cellular invasion by Cryptosporidium parvum.
Cryptosporidium parvum invasion of epithelial cells involves host cell membrane alterations which require a remodeling of the host cell actin cytoskeleton. In addition, an actin plaque, possibly associated with the dense-band region, forms within the host cytoplasm at the host-parasite interface. Here we show that Cdc42 and RhoA, but not Rac1, members of the Rho family of GTPases, are recruited to the host-parasite interface in an in vitro model of human biliary cryptosporidiosis. Interestingly, activation of Cdc42, but not RhoA, was detected in the infected cells. Neural Wiskott-Aldrich syndrome protein (N-WASP) and p34-Arc, actin-regulating downstream effectors of Cdc42, were also recruited to the host-parasite interface. Whereas cellular expression of a constitutively active mutant of Cdc42 promoted C. parvum invasion, overexpression of a dominant negative mutant of Cdc42, or depletion of Cdc42 mRNA by short interfering RNA-mediated gene silencing, inhibited C. parvum invasion. Expression of the WA fragment of N-WASP to block associated actin polymerization also inhibited C. parvum invasion. Moreover, inhibition of host cell Cdc42 activation by dominant negative mutation inhibited C. parvum-associated actin remodeling, membrane protrusion, and dense-band formation. In contrast, treatment of cells with a Rho inhibitor, exoenzyme C3, or cellular overexpression of dominant negative mutants of RhoA and Rac1 had no effect on C. parvum invasion. These data suggest that C. parvum invasion of target epithelia results from the organism's ability to activate a host cell Cdc42 GTPase signaling pathway to induce host cell actin remodeling at the attachment site.


The influence of cyclosporin A on alloantibody responses in inbred rats: provisional evidence for a serum factor with antiidiotypic activity.
The effect of cyclosporin A (CsA) on alloantibody synthesis has been investigated in inbred F344 (RTl1v1) rats receiving weekly transfusions of DA (RT1a) rat whole blood. Whereas repeated transfusion resulted in a persistent alloantibody response (Group I) administration of CsA (15 mg/kg/day) from either days 0-7 (Group II), days 8-49 (Group III) or days 15-49 (Group IV) resulted in the eventual suppression of alloantibody responses before the end of the experiment on day 49. Antiidiotypic activity was detected in sera obtained on day 49 from animals in Groups II, III and IV, shown to reside in the serum fraction of apparent molecular mass of between 150 and 170 kD and to be specific for alloantisera raised in F344 and the closely related LEW (RTl1) rats but not the unrelated AO (RTlu) strain. These experiments suggest that the immunosuppressive action of CsA may, in part, be due to the development of anti-idiotypic activity whose nature remains to be more fully characterized.


Honey and wound dehiscence: a study of surgical wounds in the mandibular bed.
There is a paucity of literature on the use of honey in wound healing after oral and maxillofacial surgical procedures. To evaluate the efficacy of the healing properties of Obudu honey in patients who developed wound dehiscence after segmental mandibular resections. This was a prospective study of 72 patients who had benign lesions of the mandible, and were treated by segmental mandibular resection, with the surgical wounds developing dehiscence. The subjects were randomized into two treatment groups of A (control, n = 36) and B (experimental, n = 36). Unlike the control, the wounds in the experimental group were dressed in honey after debridement. The ages of the patients ranged from 21 to 73 years with a mean age of 46.3 ± 2.1 years while the male: female ratio was 3:1. There was no significant demographic difference about age ( P = 0.44) and gender (P = 0.38) between the two groups. The smaller the initial circumference of the surgical wound, the shorter the duration of healing and this was significant (P = 0.001) in either of the treatment groups. Numerically, more healing was completed in the first 5 weeks in subjects in the experimental group (n = 19, 52.8%) than the control (n = 13, 36.1%). However, the duration of healing between the subjects in the control and experimental groups at the end of 9 weeks showed no significant difference (P = 0.23). Honey speeds up the healing of dehiscence wounds of resected mandible when used as dressing more than the control.


Automatic quantitative analysis of morphology of apoptotic HL-60 cells.
Morphological identification is a widespread procedure to assess the presence of apoptosis by visual inspection of the morphological characteristics or the fluorescence images. The procedure is lengthy and results are observer dependent. A quantitative automatic analysis is objective and would greatly help the routine work. We developed an image processing and segmentation method which combined the Otsu thresholding and morphological operators for apoptosis study. An automatic determination method of apoptotic stages of HL-60 cells with fluorescence images was developed. Comparison was made between normal cells, early apoptotic cells and late apoptotic cells about their geometric parameters which were defined to describe the features of cell morphology. The results demonstrated that the parameters we chose are very representative of the morphological characteristics of apoptotic cells. Significant differences exist between the cells in different stages, and automatic quantification of the differences can be achieved.


Activated Regulatory T-Cells, Dysfunctional and Senescent T-Cells Hinder the Immunity in Pancreatic Cancer.
Pancreatic cancer has one of the worst prognoses of any human malignancy and leukocyte infiltration is a major prognostic marker of the disease. As current immunotherapies confer negligible survival benefits, there is a need to better characterise leukocytes in pancreatic cancer to identify better therapeutic strategies. In this study, we analysed 32 human pancreatic cancer patients from two independent cohorts. A multi-parameter mass-cytometry analysis was performed on 32,000 T-cells from eight patients. Single-cell RNA sequencing dataset analysis was performed on a cohort of 24 patients. Multiplex immunohistochemistry imaging and spatial analysis were performed to map immune infiltration into the tumour microenvironment. Regulatory T-cell populations demonstrated highly immunosuppressive states with high TIGIT, ICOS and CD39 expression. CD8+ T-cells were found to be either in senescence or an exhausted state. The exhausted CD8 T-cells had low PD-1 expression but high TIGIT and CD39 expression. These findings were corroborated in an independent pancreatic cancer single-cell RNA dataset. These data suggest that T-cells are major players in the suppressive microenvironment of pancreatic cancer. Our work identifies multiple novel therapeutic targets that should form the basis for rational design of a new generation of clinical trials in pancreatic ductal adenocarcinoma.


Ionotropic glutamate receptors of amacrine cells of the mouse retina.
The mammalian retina contains approximately 30 different morphological types of amacrine cells, receiving glutamatergic input from bipolar cells. In this study, we combined electrophysiological and pharmacological techniques in order to study the glutamate receptors expressed by different types of amacrine cells. Whole-cell currents were recorded from amacrine cells in vertical slices of the mouse retina. During the recordings the cells were filled with Lucifer Yellow/Neurobiotin allowing classification as wide-field or narrow-field amacrine cells. Amacrine cell recordings were also carried out in a transgenic mouse line whose glycinergic amacrine cells express enhanced green fluorescent protein (EGFP). Agonist-induced currents were elicited by exogenous application of NMDA, AMPA, and kainate (KA) while holding cells at -75 mV. Using a variety of specific agonists and antagonists (NBQX, AP5, cyclothiazide, GYKI 52466, GYKI 53655, SYM 2081) responses mediated by AMPA, KA, and NMDA receptors could be dissected. All cells (n = 300) showed prominent responses to non-NMDA agonists. Some cells expressed AMPA receptors exclusively and some cells expressed KA receptors exclusively. In the majority of cells both receptor types could be identified. NMDA receptors were observed in about 75% of the wide-field amacrine cells and in less than half of the narrow-field amacrine cells. Our results confirm that different amacrine cell types express distinct sets of ionotropic glutamate receptors, which may be critical in conferring their unique temporal responses to this diverse neuronal class.


Variation of Cesium in the Ocean.
Samples of sea water from several oceans and several depths were analyzed for natural cesium by flame photometry with precision of about 3.5 percent. The average of 16 assays of surface water from four oceans was 0.37 microgram per liter, somewhat less than the average previously reported. The average cesium concentration in samples taken between 500 and 1500 meters was about 14 percent higher than the surface average, suggesting that cesium may be transported downward by particulate material. Natural cesiumn is now somewhat easier to study in the ocean than radiocesium from fallout is, and information on one complements the other.


12-hydroperoxyeicosatetraenoic acid inhibits main platelet functions by activation of soluble guanylate cyclase.
12-Hydroperoxy-5,8,10,14-eicosatetraenoic acid (12-HPETE) as well as several other fatty acid hydroperoxides are potent inhibitors of platelet activation. 12-HPETE but not 12-hydroxy-5,8,10,14-eicosatetraenoic acid blocks the U46619- and the thrombin-triggered aggregation of aspirin-treated platelets, dose dependently. 12-HPETE suppresses thromboxane production by inhibiting platelet cyclooxygenase and stimulates its own production by increasing lipoxygenase activity, although this effect does not explain the inhibitory activity of 12-HPETE during the initial phase of cell activation. The inhibitory effect is related to altered calcium homeostasis during platelet activation. 12-HPETE inhibits calcium release from intracellular stores and modifies the influx of extracellular calcium. The inhibitory effect on calcium mobilization is explained by activation of soluble guanylate cyclase. These inhibitory properties are shared by sodium nitroprusside, a compound known to activate soluble guanylate cyclase. Fatty acid hydroperoxides, especially 12-HPETE, produce a rapid and dose-dependent activation of soluble guanylate cyclase, using intact human platelets as a detection system. Activation of the enzyme shows a position isomer specificity, with 12-HPETE being the most potent activator. The generation of the labile lipoxygenase product 12-HPETE during platelet activation may modulate platelet reactivity by increasing cyclic GMP. This pathway may contribute to a physiological feedback mechanism to limit the size of a growing platelet plug.


The effect of weekend versus weekday admission on outcomes of esophageal variceal hemorrhage.
Hospital staffing is often lower on weekends than weekdays, and may contribute to higher mortality in patients admitted on weekends. Because esophageal variceal hemorrhage (EVH) requires complex management and urgent endoscopic intervention, limitations in physician expertise and the availability of endoscopy on weekends may be associated with increased EVH mortality. To assess the differences in mortality, hospital length of stay (LOS), and costs between patients admitted on weekends versus patients who were admitted on weekdays. The United States Nationwide Inpatient Sample database was used to identify patients hospitalized for EVH between 1998 and 2005. Differences in mortality, LOS, and costs between patients admitted on weekends and weekdays were evaluated using regression models with adjustment for patient and clinical factors, including the timing of endoscopy. Between 1998 and 2005, 36,734 EVH admissions to 2207 hospitals met the inclusion criteria. Compared with patients admitted on weekdays, individuals admitted on the weekend were slightly less likely to undergo endoscopy on the day of admission (45% versus 43%, respectively; P=0.01) and by the second day (81% versus 75%; P<0.0001). However, mortality (11.3% versus 10.8%; P=0.20) and the requirement for endoscopic therapy (70% versus 69%; P=0.08) or portosystemic shunt insertion (4.4% versus 4.7%; P=0.32) did not differ between weekend and weekday admissions. After adjusting for confounding factors, including the timing of endoscopy, the risk of mortality was similar between weekend and weekday admissions (OR 1.05; 95% CI 0.97 to 1.14). Although LOS was similar between groups, adjusted hospital charges were 4.0% greater (95% CI 2.3 to 5.8%) for patients hospitalized on the weekend. In patients with EVH, admission on the weekend is associated with a small delay in receiving endoscopic intervention, but no difference in mortality or the requirement for portosystemic shunt insertion. The weekend effect observed for some medical and surgical conditions does not apply to patients with EVH.


The antinatriuretic action of biosynthetic human growth hormone in man involves activation of the renin-angiotensin system.
Previous studies using human pituitary extracts have not resolved whether the sodium retaining effects of human growth hormone (hGH) are mediated in part by increased aldosterone secretion. We have studied the effects of an authentic biosynthetic GH (bio-hGH) preparation on sodium metabolism and on the activity of the renin-angiotensin system. Six young men were administered this preparation at 0.2 U/kg/d subcutaneously for five consecutive days. Twenty-four-hour urine collections were obtained for measurement of sodium excretion and osmolality and blood collected for quantitating changes in sodium, osmolality, plasma renin activity (PRA), aldosterone, and arginine vasopressin (AVP) concentrations. Bio-hGH administration resulted in a fall in 24-hour urinary sodium excretion (197 +/- 38 to 42 +/- 20 mmol, mean +/- SD, P less than .005), a reduction in urine volume (1,652 +/- 182 to 848 +/- 348 mL, P less than .05) but not osmolality. PRA increased significantly from 1,118 +/- 73 to 3,608 +/- 1,841 fmol angiotensin 1 L/s (P less than .005), which was associated with a sevenfold increase in plasma aldosterone concentration (52 +/- 12 to 402 +/- 99 pg/mL, P less than .001). Plasma osmolality and AVP concentrations did not change significantly. The results show that Bio-GH-induced retention of sodium involves the activation of the renin-angiotensin system. This mechanism may explain in part the occurrence of plasma volume expansion and hypertension in acromegaly and suggests a risk of fluid retention and possibly hypertension in subjects receiving supraphysiological doses of bio-hGH for treatment of short stature.


Duplicated antagonistic EPF peptides optimize grass stomatal initiation.
Peptide signaling has emerged as a key component of plant growth and development, including stomatal patterning, which is crucial for plant productivity and survival. Although exciting progress has been made in understanding EPIDERMAL PATTERNING FACTOR (EPF) signaling in Arabidopsis, the mechanisms by which EPF peptides control different stomatal patterns and morphologies in grasses are poorly understood. Here, by examining expression patterns, overexpression transgenics and cross-species complementation, the antagonistic stomatal ligands orthologous to Arabidopsis AtEPF2 and AtSTOMAGEN/AtEPFL9 peptides were identified in Triticum aestivum (wheat) and the grass model organism Brachypodium distachyon. Application of bioactive BdEPF2 peptides inhibited stomatal initiation, but not the progression or differentiation of stomatal precursors in Brachypodium. Additionally, the inhibitory roles of these EPF peptides during grass stomatal development were suppressed by the contrasting positive action of the BdSTOMAGEN peptide in a dose-dependent manner. These results not only demonstrate how conserved EPF peptides that control different stomatal patterns exist in nature, but also suggest new strategies to improve crop yield through the use of plant-derived antagonistic peptides that optimize stomatal density on the plant epidermis.


Evaluation of phenolic profile, antioxidant and anticancer potential of two main representants of Zingiberaceae family against B164A5 murine melanoma cells.
Curcuma longa Linnaeus and Zingiber officinale Roscoe are two main representatives of Zingiberaceae family studied for a wide range of therapeutic properties, including: antioxidant, anti-inflammatory, anti-angiogenic, antibacterial, analgesic, immunomodulatory, proapoptotic, anti-human immunodeficiency virus properties and anticancer effects. This study was aimed to analyse the ethanolic extracts of Curcuma rhizome (Curcuma longa Linnaeus) and Zingiber rhizome (Zingiber officinale Roscoe) in terms of polyphenols, antioxidant activity and anti-melanoma potential employing the B164A5 murine melanoma cell line. In order to evaluate the total content of polyphenols we used Folin-Ciocâlteu method. The antioxidant activity of the two ethanolic extracts was determined by DPPH assay, and for the control of antiproliferative effect it was used MTT proliferation assay, DAPI staining and Annexin-FITC-7AAD double staining test. Results showed increased polyphenols amount and antioxidant activity for Curcuma rhizome ethanolic extract. Moreover, 100 μg/ml of ethanolic plant extract from both vegetal products presented in a different manner an antiproliferative, respectively a proapoptotic effect on the selected cell line. The study concludes that Curcuma rhizome may be a promising natural source for active compounds against malignant melanoma.


Incidence of Clostridium difficile in infants in rural and urban areas of Nigeria.
Stool specimens collected from 320 infants under 5 years of age resident in both rural and urban centres of Nsukka and Enugu towns in Anambra State, Nigeria were examined for the presence of Clostridium difficile and the frequency of occurrence determined. Clostridium difficile was isolated from 156 out of 320 samples (48.8%). The frequency was higher in infants resident in rural areas (52.5-48.3%) than in their urban counterparts (47.8-42%). A high frequency rate (77%) was obtained in diarrheal cases as against 42% in non diarrheal cases. A frequency of 57.7% was obtained in infants in the age group of 1 day - 1 yr, which decline with age to the lowest 5.6% in the age group of 4.5 years. The frequency of 66.1% was obtained in infants breast-fed plus formula supplementation, 41.7% in infants exclusively breast-fed and 33% in infants fed exclusively with formula foods. The findings suggest that the type of living environments, together with age and mode of feeding may contribute to the high intestinal carriage of Clostridium difficile in infants in parts of Nigeria.


Thalamic innervation of the direct and indirect basal ganglia pathways in the rat: Ipsi- and contralateral projections.
The present study describes the thalamic innervation coming from the rat parafascicular nucleus (PF) onto striatal and subthalamic efferent neurons projecting either to the globus pallidus (GP) or to the substantia nigra pars reticulata (SNr) by using a protocol for multiple neuroanatomical tracing. Both striatofugal neurons targeting the ipsilateral SNr (direct pathway) as well as striatal efferent neurons projecting to the ipsilateral GP (indirect pathway) were located within the terminal fields of the thalamostriatal afferents. In the subthalamic nucleus (STN), both neurons projecting to ipsilateral GP as well as neurons projecting to ipsilateral SNr also appear to receive thalamic afferents. Although the projections linking the caudal intralaminar nuclei with the ipsilateral striatum and STN are far more prominent, we also noticed that thalamic axons could gain access to the contralateral STN. Furthermore, a small number of STN neurons were seen to project to both the contralateral GP and PF nuclei. These ipsi- and contralateral projections enable the caudal intralaminar nuclei to modulate the activity of both the direct and the indirect pathway.


Capturing Provenance, Evolution and Modification of Clinical Protocols via a Heterogeneous, Semantic Social Network.
Healthcare delivery is largely based on medical best practices as in clinical protocols. Research so far has addressed the computerized execution of clinical protocols by developing a number of related representation languages, execution engines and integrated platforms to support real time execution. However, much less effort has been put into organizing clinical protocols for use and reuse. In this paper we propose a heterogeneous semantic social network to describe and organize clinical protocols based on their provenance, evolution and modifications. The proposed approach allows semantic tagging and enrichment of clinical protocols so that they can be used and re-used across platforms and also be linked directly to other relevant scientific information, e.g. published works in PubMed or personal health records, and other clinical information systems.


Multiple endocrine neoplasia type 1 knockout mice develop parathyroid, pancreatic, pituitary and adrenal tumours with hypercalcaemia, hypophosphataemia and hypercorticosteronaemia.
Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant disorder characterized in man by parathyroid, pancreatic, pituitary and adrenal tumours. The MEN1 gene encodes a 610-amino acid protein (menin) which is a tumour suppressor. To investigate the in vivo role of menin, we developed a mouse model, by deleting Men1 exons 1 and 2 and investigated this for MEN1-associated tumours and serum abnormalities. Men1(+/-) mice were viable and fertile, and 220 Men1(+/-) and 94 Men1(+/+) mice were studied between the ages of 3 and 21 months. Survival in Men1(+/-) mice was significantly lower than in Men1(+/+) mice (<68% vs >85%, P<0.01). Men1(+/-) mice developed, by 9 months of age, parathyroid hyperplasia, pancreatic tumours which were mostly insulinomas, by 12 months of age, pituitary tumours which were mostly prolactinomas, and by 15 months parathyroid adenomas and adrenal cortical tumours. Loss of heterozygosity and menin expression was demonstrated in the tumours, consistent with a tumour suppressor role for the Men1 gene. Men1(+/-) mice with parathyroid neoplasms were hypercalcaemic and hypophosphataemic, with inappropriately normal serum parathyroid hormone concentrations. Pancreatic and pituitary tumours expressed chromogranin A (CgA), somatostatin receptor type 2 and vascular endothelial growth factor-A. Serum CgA concentrations in Men1(+/-) mice were not elevated. Adrenocortical tumours, which immunostained for 3-beta-hydroxysteroid dehydrogenase, developed in seven Men1(+/-) mice, but resulted in hypercorticosteronaemia in one out of the four mice that were investigated. Thus, these Men1(+/-) mice are representative of MEN1 in man, and will help in investigating molecular mechanisms and treatments for endocrine tumours.


Effects of a flotation vest and water flow rate on limb kinematics of Siberian Huskies swimming against a current.
To evaluate the effects of a flotation vest (FV) and water flow rate (WFR) on limb kinematics of dogs swimming against a current. 7 (1 male and 6 female) healthy adult Siberian Huskies. Dogs were habituated to swim with and without an FV beside an investigator in a continuous-flow pool against WFRs up to 2.9 km/h. During each of 4 experimental sessions in a repeated-measures study, markers were wrapped around the right carpus and tarsus, and a video was recorded while each dog swam with or without an FV for about 2 minutes at each of 7 WFRs between 0 and 2.9 km/h when the WFR was incrementally decreased or increased. Motion tracking software was used to measure stroke excursion and frequency. Stroke excursion varied more than frequency among all dogs and in response to changes in experimental conditions. The male dog and 1 female dog were unable to complete the study. For the remaining 5 dogs across all experimental conditions, mean tarsus excursion was 30% that of the carpus. Mean total excursion (sum of the excursion-frequency products for the carpus and tarsus) decreased when an FV was worn and increased with WFR by 69% and 19% when WFR was incrementally increased and decreased, respectively. In dogs, range of motion during swimming was greater for the carpus than tarsus, when an FV was not worn, and increased more with WFR when WFR was incrementally increased. Those factors should be considered during swimming-based rehabilitation.


Cost-effectiveness analysis of risk-factor guided and birth-cohort screening for chronic hepatitis C infection in the United States.
No consensus exists on screening to detect the estimated 2 million Americans unaware of their chronic hepatitis C infections. Advisory groups differ, recommending birth-cohort screening for baby boomers, screening only high-risk individuals, or no screening. We assessed one-time risk assessment and screening to identify previously undiagnosed 40-74 year-olds given newly available hepatitis C treatments. A Markov model evaluated alternative risk-factor guided and birth-cohort screening and treatment strategies. Risk factors included drug use history, blood transfusion before 1992, and multiple sexual partners. Analyses of the National Health and Nutrition Examination Survey provided sex-, race-, age-, and risk-factor-specific hepatitis C prevalence and mortality rates. Nine strategies combined screening (no screening, risk-factor guided screening, or birth-cohort screening) and treatment (standard therapy-peginterferon alfa and ribavirin, Interleukin-28B-guided (IL28B) triple-therapy-standard therapy plus a protease inhibitor, or universal triple therapy). Response-guided treatment depended on HCV genotype. Outcomes include discounted lifetime costs (2010 dollars) and quality adjusted life-years (QALYs). Compared to no screening, risk-factor guided and birth-cohort screening for 50 year-olds gained 0.7 to 3.5 quality adjusted life-days and cost $168 to $568 per person. Birth-cohort screening provided more benefit per dollar than risk-factor guided screening and cost $65,749 per QALY if followed by universal triple therapy compared to screening followed by IL28B-guided triple therapy. If only 10% of screen-detected, eligible patients initiate treatment at each opportunity, birth-cohort screening with universal triple therapy costs $241,100 per QALY. Assuming treatment with triple therapy, screening all individuals aged 40-64 years costs less than $100,000 per QALY. The cost-effectiveness of one-time birth-cohort hepatitis C screening for 40-64 year olds is comparable to other screening programs, provided that the healthcare system has sufficient capacity to deliver prompt treatment and appropriate follow-on care to many newly screen-detected individuals.