Determination of silver nanoparticle release from antibacterial fabrics into artificial sweat.
Silver nanoparticles have been used in numerous commercial products, including textiles, to prevent bacterial growth. Meanwhile, there is increasing concern that exposure to these nanoparticles may cause potential adverse effects on humans as well as the environment. This study determined the quantity of silver released from commercially claimed nanosilver and laboratory-prepared silver coated fabrics into various formulations of artificial sweat, each made according to AATCC, ISO and EN standards. For each fabric sample, the initial amount of silver and the antibacterial properties against the model Gram-positive (S. aureus) and Gram-negative (E. coli) bacteria on each fabric was investigated. The results showed that silver was not detected in some commercial fabrics. Furthermore, antibacterial properties of the fabrics varied, ranging from 0% to greater than 99%. After incubation of the fabrics in artificial sweat, silver was released from the different fabrics to varying extents, ranging from 0 mg/kg to about 322 mg/kg of fabric weight. The quantity of silver released from the different fabrics was likely to be dependent on the amount of silver coating, the fabric quality and the artificial sweat formulations including its pH. This study is the unprecedented report on the release of silver nanoparticles from antibacterial fabrics into artificial sweat. This information might be useful to evaluate the potential human risk associated with the use of textiles containing silver nanoparticles.


Haploinsufficiency of a spliceosomal GTPase encoded by EFTUD2 causes mandibulofacial dysostosis with microcephaly.
Mandibulofacial dysostosis with microcephaly (MFDM) is a rare sporadic syndrome comprising craniofacial malformations, microcephaly, developmental delay, and a recognizable dysmorphic appearance. Major sequelae, including choanal atresia, sensorineural hearing loss, and cleft palate, each occur in a significant proportion of affected individuals. We present detailed clinical findings in 12 unrelated individuals with MFDM; these 12 individuals compose the largest reported cohort to date. To define the etiology of MFDM, we employed whole-exome sequencing of four unrelated affected individuals and identified heterozygous mutations or deletions of EFTUD2 in all four. Validation studies of eight additional individuals with MFDM demonstrated causative EFTUD2 mutations in all affected individuals tested. A range of EFTUD2-mutation types, including null alleles and frameshifts, is seen in MFDM, consistent with haploinsufficiency; segregation is de novo in all cases assessed to date. U5-116kD, the protein encoded by EFTUD2, is a highly conserved spliceosomal GTPase with a central regulatory role in catalytic splicing and post-splicing-complex disassembly. MFDM is the first multiple-malformation syndrome attributed to a defect of the major spliceosome. Our findings significantly extend the range of reported spliceosomal phenotypes in humans and pave the way for further investigation in related conditions such as Treacher Collins syndrome.


Temperature profiles of 980- and 1,470-nm endovenous laser ablation, endovenous radiofrequency ablation and endovenous steam ablation.
Endovenous thermal ablation (EVTA) techniques are very effective for the treatment of varicose veins, but their exact working mechanism is still not well documented. The lack of knowledge of mechanistic properties has led to a variety of EVTA protocols and a commercially driven dissemination of new or modified techniques without robust scientific evidence. The aim of this study is to compare temperature profiles of 980-and 1,470-nm endovenous laser ablation (EVLA), segmental radiofrequency ablation (RFA), and endovenous steam ablation (EVSA). In an experimental setting, temperature measurements were performed using thermocouples; raw potato was used to mimic a vein wall. Two laser wavelengths (980 and 1,470 nm) were used with tulip-tip fibers and 1,470 nm also with a radial-emitting fiber. Different powers and pullback speeds were used to achieve fluences of 30, 60, and 90 J/cm. For segmental RFA, 1 cycle of 20 s was analyzed. EVSA was performed with two and three pulses of steam per centimeter. Maximum temperature increase, time span of relevant temperature increase, and area under the curve of the time of relevant temperature increase were measured. In all EVLA settings, temperatures increased and decreased rapidly. High fluence is associated with significantly higher temperatures and increased time span of temperature rise. Temperature profiles of 980- and 1,470-nm EVLA with tulip-tip fibers did not differ significantly. Radial EVLA showed significantly higher maximum temperatures than tulip-tip EVLA. EVSA resulted in mild peak temperatures for longer durations than EVLA. Maximum temperatures with three pulses per centimeter were significantly higher than with two pulses. RFA temperature rises were relatively mild, resulting in a plateau-shaped temperature profile, similar to EVSA. Temperature increase during EVLA is fast with a high-peak temperature for a short time, where EVSA and RFA have longer plateau phases and lower maximum temperatures.


Control of thermoregulatory sweating during exercise in the heat.
The purposes of this study were the following: 1) to determine whether erythrocyte infusion alters the control of thermoregulatory sweating and 2) to demonstrate how increases and decreases of both plasma tonicity and blood volume influence the thermoregulatory control parameters of threshold temperature and sweating sensitivity. Six non-heat-acclimated and five heat-acclimated males attempted heat stress tests (HSTs) both before and shortly after (48-96 h) autologous erythrocyte infusion. The non-heat-acclimated subjects were euhydrated for both HSTs, whereas the heat-acclimated subjects were studied in a euhydrated and a hypohydrated (-5% body wt) condition both pre- and postinfusion (500 ml of solution containing approximately 60% hematocrit of autologous erythrocytes). The HSTs consisted of treadmill exercise (335 W.m-2) in a hot (35 degrees C, 45% relative humidity) environment, and esophageal temperature and local sweating rate were continuously measured during 25 min of exercise. These experiments resulted in a matrix of conditions where both plasma tonicity and blood volume were increased or decreased relative to control conditions (euhydration, preinfusion). The findings concerning thermoregulatory sweating during exercise in the heat were summarized as follows: 1) acute polycythemia decreases the threshold temperature and increases the sweating sensitivity, 2) both threshold temperature and sweating sensitivity are increased or decreased from control levels dependent on the combined influence of plasma tonicity and blood volume, and 3) equations are presented that describe how plasma tonicity and blood volume alter threshold temperature and sweating sensitivity values.


Impact of thrombus migration on clinical outcomes in patients with internal carotid artery occlusions and patent middle cerebral artery.
Patency of the middle cerebral artery (MCA) in acute ischemic stroke with internal carotid artery (ICA) occlusions is associated with less severe stroke and favorable outcomes. However, thrombus migration to distal intracranial vessels may lead to unfavorable outcomes. We investigated the influence of thrombus migration on clinical outcomes in patients with ICA occlusions and patent MCA. We retrospectively analyzed patients with acute ischemic stroke compromising ICA occlusions and patent MCA who were consecutively admitted to our hospital between January 2006 and March 2016. Thrombus migration was assessed (1) by analyzing the discrepancies in arterial occlusion sites between initial imaging and follow-up imaging and (2) by analyzing how occlusion sites changed during endovascular therapy. Thirty-eight patients (mean age: 74.9 years; 23 men, 15 women, median National Institutes of Health Stroke Scale score = 7.5) with ICA occlusions and patent MCA were ultimately included. We identified 10 patients (26%) with thrombus migration (spontaneous: 3; during endovascular therapy: 7). Patients with thrombus migration had higher rates of unfavorable functional outcomes (modified Rankin Scale scores 3-6 at 90 days) than those without thrombus migration (90% vs. 39%, p < .01). Multivariate analysis showed that thrombus migration was independently related to unfavorable functional outcomes (odds ratio, 42.9; 95% confidence interval, 1.5-1211.0; p = .03). Thrombus migration in cases of ICA occlusion with patent MCA is associated with poor prognosis. Careful monitoring is required under these conditions even if the initial clinical presentation is mild.


Dual genetic pathways of endothelin-mediated intercellular signaling revealed by targeted disruption of endothelin converting enzyme-1 gene.
Recent gene targeting studies have revealed unexpected roles for endothelins in the development of neural crest-derived tissues. Endothelin converting enzyme-1 (ECE-1) catalyzes the proteolytic activation of big endothelin-1 to endothelin-1(ET-1) in vitro. However, the importance of ECE-1 cleavage in the multiple endothelin pathways in vivo is unknown. Here we generated a targeted null mutation in the mouse ECE-1 gene. ECE-1-/- term embryos exhibited craniofacial and cardiac abnormalities virtually identical to the defects seen in ET-1 and endothelin A receptor (ETA)-deficient embryos. Epidermal melanocytes as well as enteric neurons of the distal gut were also absent in ECE-1-/- embryos, reproducing the developmental phenotype seen in ET-3-/- and endothelin B receptor (ETB)-/- mice. Surprisingly, large amounts of mature ET-1 peptide are found in ECE-1-/- embryos, indicating that non-ECE-1 protease(s) can activate ET-1 at certain sites. However, these enzymes cannot produce sufficient mature endothelin at the locations crucial for normal embryonic development. These findings reveal that ECE-1 is a bona fide activating protease for both big ET-1 and big ET-3 in vivo, and that the cell-cell communication pathways represented by the ET-1/ECE-1/ETA axis and the ET-3/ECE-1/ETB axis are each involved in the development of distinct subsets of neural crest cell lineages. Mutations in ECE-1 may cause developmental defects in humans, such as Hirschsprung disease, velocardiofacial syndrome and related neurocristopathies.


Resident cardiac progenitor cells: at the heart of regeneration.
Stem cell therapy has recently emerged as an innovative strategy over conventional cardiovascular treatments to restore cardiac function in patients affected by ischemic heart disease. Various stem cell populations have been tested and their potential for cardiac repair has been analyzed. Embryonic stem cells retain the greatest differentiation potential, but concerns persist with regard to their immunogenic and teratogenic effects. Although adult somatic stem cells are not tumourigenic and easier to use in an autologous setting, they exist in small numbers and possess reduced differentiation potential. Traditionally the heart was considered to be a post-mitotic organ; however, this dogma has recently been challenged with the identification of a reservoir of resident stem cells, defined as cardiac progenitor cells (CPCs). These endogenous progenitors may represent the best candidates for cardiovascular cell therapy, as they are tissue-specific, often pre-committed to a cardiac fate, and display a greater propensity to differentiate towards cardiovascular lineages. This review will focus on current research into the biology of CPCs and their regenerative potential. This article is part of a special issue entitled, "Cardiovascular Stem Cells Revisited".


Epigenetic control in rheumatoid arthritis synovial fibroblasts.
Rheumatoid arthritis synovial fibroblasts (RASFs) are the effector cells of cartilage and bone destruction. These cells show an 'intrinsically' activated and aggressive phenotype that results in the increased production of matrix-degrading enzymes and adhesion molecules, and is conserved over long-term passage in vitro. The three main mechanisms of epigenetic control -- DNA methylation, histone modifications and microRNA activity -- interact in the development of the RASF phenotype. The extent of global DNA methylation is reduced in synoviocytes in situ and RASFs in vitro. In addition, histone hyperacetylation occurs and specific microRNAs are expressed in RASFs. Normal synovial fibroblasts cultured in a hypomethylating milieu acquire an activated phenotype similar to that of RASFs. These findings suggest that epigenetic control, in particular the control of DNA methylation, is deficient in RASFs. Genome-wide analyses of the epigenome will enable the detection of additional genes involved in the pathogenesis of rheumatoid arthritis, the identification of epigenetic biomarkers, and potentially the development of a therapeutic regimen that targets activated RASFs.


Epitope mapping and functional properties of anti-intercellular adhesion molecule-3 (CD50) monoclonal antibodies.
Intercellular adhesion molecule-3 (ICAM-3, CD50), a member of the immunoglobulin gene superfamily, is a major ligand for the lymphocyte function-associated antigen 1 (LFA-1, CD18/CD11a) in the resting immune system and plays a role as a signaling and costimulatory molecule on T lymphocytes. In this study we have generated a large panel of anti-ICAM-3 monoclonal antibodies (mAb) and show that the biological effects of these antibodies are critically dependent on the epitope recognized. By using an adhesion assay employing COS cells expressing LFA-1 binding to recombinant chimeric ICAM-3-Fc proteins (which overcomes the confounding effects of interleukocyte LFA-1/ICAM binding events), we have been able to examine the effects of these antibodies in blocking LFA-1/ICAM-3 adhesion. Our data suggests that only a small minority of ICAM-3 mAb, recognizing a distinct epitope, are able to mimic the effects of LFA-1 binding to ICAM-3. Moreover these antibodies are functionally distinct as defined by their costimulatory activity and ability to elicit interleukin-2 production and cell proliferation in T lymphocytes.


Osteomyelitis in the spinal cord injured: a review and a preliminary report on the use of hyperbaric oxygen therapy.
Spinal Cord Injury patients are liable to develop osteomyelitis mostly by extension from pressure ulcers. In 2055 records reviewed in the Long Beach Spinal Cord Injury Service of the Veterans Administration Medical Center, the incidence was found to be 4.3 per cent. Of these osteomyelitis developed secondary to pressure ulcers in 88 per cent, the rest developed as a result of trauma and/or surgery. Forty-four patients manifesting chronic osteomyelitis were treated in a monoplace hyperbaric oxygen (HBO) chamber, in addition to receiving antibiotic and surgical treatment. HBO was found useful as an adjunct to help to resolve the bone infection and encourage wound healing. Two-thirds of the patients were cured, and the follow-up was from 6 months to 9 years. We believe that HBO is a useful adjunctive therapeutic measure in the management of chronic osteomyelitis in the spinal cord injured and in the prevention of its complications.


Microsecond Time Scale Proton Rotating-Frame Relaxation under Magic Angle Spinning.
This paper deals with the theoretical foundation of proton magic angle spinning rotating-frame relaxation (R1ρ) and establishes the range of validity and accuracy of the presented approach to describe low-amplitude microsecond time scale motion in the solid state. Beside heteronuclear dipolar and chemical shift anisotropy interactions, a major source of relaxation for protons is the homonuclear dipolar interaction. For this latter relaxation process, no general analytical equation has been published until now, which would describe the R1ρ relaxation at any spinning speed, spin-lock field, or tilt angle. To validate the derived equations, we compared the analytical relaxation rates, obtained by solving the master equation within the framework of Redfield theory, with numerically simulated relaxation rates. We found that for small opening angles (∼10°), the relaxation rates obtained with stochastic Liouville simulations agree well with the analytical Redfield relaxation rates for a large range of motional correlation times. However, deviations around the rotary-resonance conditions highlight the fact that Redfield treatment of the solid-state relaxation rates can only provide qualitative insights into the microsecond time scale motion.


Loss of CCR5 results in glucose intolerance in diet-induced obese mice.
Macrophage and T cell infiltration into metabolic tissues contributes to obesity-associated inflammation and insulin resistance (IR). C-C chemokine receptor 5 (CCR5), expressed on macrophages and T cells, plays a critical role in the recruitment and activation of proinflammatory M1 and TH1 immune cells to tissues and is elevated in adipose tissue (AT) and liver of obese humans and mice. Thus, we hypothesized that deficiency of CCR5 would protect against diet-induced inflammation and IR. CCR5-deficient (CCR5(-/-)) mice and C57BL/6 (WT) controls were fed 10% low-fat (LF) or 60% high-fat (HF) diets for 16 wk. HF feeding increased adiposity, blood glucose, and plasma insulin levels equally in both genotypes. Opposing our hypothesis, HF-fed CCR5(-/-) mice were significantly more glucose intolerant than WT mice. In AT, there was a significant reduction in the M1-associated gene CD11c, whereas M2 associated genes were not different between genotypes. In addition, HF feeding caused a twofold increase in CD4(+) T cells in the AT of CCR5(-/-) compared with WT mice. In liver and muscle, no differences in immune cell infiltration or inflammatory cytokine expression were detected. However, in AT and muscle, there was a mild reduction in insulin-induced phosphorylation of AKT and IRβ in CCR5(-/-) compared with WT mice. These findings suggest that whereas CCR5 plays a minor role in regulating immune cell infiltration and inflammation in metabolic tissues, deficiency of CCR5 impairs systemic glucose tolerance as well as AT and muscle insulin signaling.


Efficient derivation of multipotent neural stem/progenitor cells from non-human primate embryonic stem cells.
The common marmoset (Callithrix jacchus) is a small New World primate that has been used as a non-human primate model for various biomedical studies. We previously demonstrated that transplantation of neural stem/progenitor cells (NS/PCs) derived from mouse and human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) promote functional locomotor recovery of mouse spinal cord injury models. However, for the clinical application of such a therapeutic approach, we need to evaluate the efficacy and safety of pluripotent stem cell-derived NS/PCs not only by xenotransplantation, but also allotransplantation using non-human primate models to assess immunological rejection and tumorigenicity. In the present study, we established a culture method to efficiently derive NS/PCs as neurospheres from common marmoset ESCs. Marmoset ESC-derived neurospheres could be passaged repeatedly and showed sequential generation of neurons and astrocytes, similar to that of mouse ESC-derived NS/PCs, and gave rise to functional neurons as indicated by calcium imaging. Although marmoset ESC-derived NS/PCs could not differentiate into oligodendrocytes under default culture conditions, these cells could abundantly generate oligodendrocytes by incorporating additional signals that recapitulate in vivo neural development. Moreover, principal component analysis of microarray data demonstrated that marmoset ESC-derived NS/PCs acquired similar gene expression profiles to those of fetal brain-derived NS/PCs by repeated passaging. Therefore, marmoset ESC-derived NS/PCs may be useful not only for accurate evaluation by allotransplantation of NS/PCs into non-human primate models, but are also applicable to analysis of iPSCs established from transgenic disease model marmosets.


Vocational Community College Students' Conversations about Binge Drinking.
The large amount of vocational community college students that continue showing binge-drinking behavior might be an indication that interventions targeting this behavior have not been hugely successful. It might be that these interventions targeted beliefs that are less or not related with vocational community college binge-drinking behavior. The aim of this study was to identify vocational community college students` salient beliefs about binge drinking. In the context of the integrative model of behavioral prediction, we applied deductive qualitative content analysis of conversations about binge drinking. The analysis of vocational community college students` conversations about binge drinking revealed a significant amount of salient beliefs, such as cognitive and affective outcome beliefs and efficacy beliefs. These beliefs may be important indicators of vocational community college students` binge drinking. Moreover, to our knowledge, this study was the first to reveal a new set of beliefs, namely social judgment beliefs (i.e., vocational community college students` judgments about their peers` binge-drinking behavior). We believe that our study yielded salient beliefs that may serve as input for future interventions targeting binge drinking among vocational community college students.


Constitutive expression of interferon-induced human MxA protein in transgenic tobacco plants does not confer resistance to a variety of RNA viruses.
MxA is a key component in the interferon-induced antiviral defense in humans. After viral infections, MxA is rapidly induced and accumulates in the cytoplasm. The multiplication of many RNA viruses, including all bunyaviruses tested so far, is inhibited by MxA. These findings prompted us to express MxA in plants in an attempt to create resistance to tospoviruses. Here, we report the generation of transgenic tobacco plants that constitutively express MxA under the control of the 35 S cauliflower mosaic virus promotor. Northern and western blot analysis confirmed the expression of MxA in several transgenic plant lines. MxA expression had no obvious detrimental effects on plant growth and fertility. However, challenge experiments with tomato spotted wilt virus, tomato chlorotic spot virus, and groundnut ringspot virus revealed no increased resistance of MxA-transgenic tobacco plants to tospovirus infections. Neither was the multiplication of tobacco mosaic virus, cucumber mosaic virus and potato virus Y inhibited in MxA-transgenic plants. The results indicate that the expression of human MxA alone does not enhance virus resistance in planta.


Oscillatory patterns in temporal lobe reveal context reinstatement during memory search.
Psychological theories of memory posit that when people recall a past event, they not only recover the features of the event itself, but also recover information associated with other events that occurred nearby in time. The events surrounding a target event, and the thoughts they evoke, may be considered to represent a context for the target event, helping to distinguish that event from similar events experienced at different times. The ability to reinstate this contextual information during memory search has been considered a hallmark of episodic, or event-based, memory. We sought to determine whether context reinstatement may be observed in electrical signals recorded from the human brain during episodic recall. Analyzing electrocorticographic recordings taken as 69 neurosurgical patients studied and recalled lists of words, we uncovered a neural signature of context reinstatement. Upon recalling a studied item, we found that the recorded patterns of brain activity were not only similar to the patterns observed when the item was studied, but were also similar to the patterns observed during study of neighboring list items, with similarity decreasing reliably with positional distance. The degree to which individual patients displayed this neural signature of context reinstatement was correlated with their tendency to recall neighboring list items successively. These effects were particularly strong in temporal lobe recordings. Our findings show that recalling a past event evokes a neural signature of the temporal context in which the event occurred, thus pointing to a neural basis for episodic memory.


Accessibility of microRNA binding sites in metastable RNA secondary structures in the presence of SNPs.
We study microRNA (miRNA) bindings to metastable RNA secondary structures close to minimum free energy conformations in the context of single nucleotide polymorphisms (SNPs) and messenger RNA (mRNA) concentration levels, i.e. whether features of miRNA bindings to metastable conformations could provide additional information supporting the differences in expression levels of the two sequences defined by a SNP. In our study, the instances [mRNA/3'UTR; SNP; miRNA] were selected based on strong expression level analyses, SNP locations within binding regions and the computationally feasible identification of metastable conformations. We identified 14 basic cases [mRNA; SNP; miRNA] of 3' UTR-lengths ranging from 124 up to 1078 nt reported in recent literature, and we analyzed the number, structure and miRNA binding to metastable conformations within an energy offset above mfe conformations. For each of the 14 instances, the miRNA binding characteristics are determined by the corresponding STarMir output. Among the different parameters we introduced and analyzed, we found that three of them, related to the average depth and average opening energy of metastable conformations, may provide supporting information for a stronger separation between miRNA bindings to the two alleles defined by a given SNP. At http://kks.inf.kcl.ac.uk/MSbind.html the MSbind tool is available for calculating features of metastable conformations determined by putative miRNA binding sites.


Removal of pharmaceuticals in pre-denitrifying MBBR - Influence of organic substrate availability in single- and three-stage configurations.
Due to the limited efficiency of conventional biological treatment, innovative solutions are being explored to improve the removal of trace organic chemicals in wastewater. Controlling biomass exposure to growth substrate represents an appealing option for process optimization, as substrate availability likely impacts microbial activity, hence organic trace chemical removal. This study investigated the elimination of pharmaceuticals in pre-denitrifying moving bed biofilm reactors (MBBRs), where biofilm exposure to different organic substrate loading and composition was controlled by reactor staging. A three-stage MBBR and a single-stage reference MBBR (with the same operating volume and filling ratio) were operated under continuous-flow conditions (18 months). Two sets of batch experiments (day 100 and 471) were performed to quantify and compare pharmaceutical removal and denitrification kinetics in the different MBBRs. Experimental results revealed the possible influence of retransformation (e.g., from conjugated metabolites) and enantioselectivity on the removal of selected pharmaceuticals. In the second set of experiments, specific trends in denitrification and biotransformation kinetics were observed, with highest and lowest rates/rate constants in the first (S1) and the last (S3) staged sub-reactors, respectively. These observations were confirmed by removal efficiency data obtained during continuous-flow operation, with limited removal (<10%) of recalcitrant pharmaceuticals and highest removal in S1 within the three-stage MBBR. Notably, biotransformation rate constants obtained for non-recalcitrant pharmaceuticals correlated with mean specific denitrification rates, maximum specific growth rates and observed growth yield values. Overall, these findings suggest that: (i) the long-term exposure to tiered substrate accessibility in the three-stage configuration shaped the denitrification and biotransformation capacity of biofilms, with significant reduction under substrate limitation; (ii) biotransformation of pharmaceuticals may have occurred as a result of cometabolism by heterotrophic denitrifying bacteria.


The production performance of imported Friesian cattle under heavy Trypanosoma challenge in a rain forest zone of Nigeria.
The productivity of 76 newly imported pregnant Friesian cattle and two bulls under heavy trypanosome challenge in the rain forest belt of Nigeria is reported. At the first visit in August 1989 and within 7 months of arrival of the heifers, the herd population had reduced by 26 (33.3%) as a result of deaths (six animals) and culls/salvages (20). The surviving 52 animals were generally in poor health with classical symptoms of trypanosomiasis. Thirty-one (40.8%) of the pregnancies were unsuccessful because of abortions (13 animals), premature births (seven), embryonic deaths (five) and death of heifers (six). Of the 45 successful calvings, 16 perinatal deaths occurred. All serum samples were negative for brucellosis. Only 41 (63.1%) of the 65 productive heifers lactated of which 24 (58.5%) yielded milk only for 6 months and less. The remaining 17 (41.5%) heifers were still at different stages of lactation ranging from 3 to 7 months within the period of analysis. Treatment with isometamidium (Samorin) at 0.5 mg/kg body weight cured the infection and prevented reinfections and/or relapses within 3 months of administration. A rise in the haematocrit and milk production after Samorin treatment was recorded. Careful analysis of the outbreak indicated that the reproductive wastages and poor lactational performance may have been induced by the severe trypanosomiasis diagnosed in the herd.


Cloning and nucleotide sequence of the gene encoding the positive regulator (DmpR) of the phenol catabolic pathway encoded by pVI150 and identification of DmpR as a member of the NtrC family of transcriptional activators.
The catabolic plasmid pVI150 of Pseudomonas sp. strain CF600 encodes all the genetic information required for the regulated metabolism of phenol and some of its methyl-substituted derivatives. The structural dmp genes of the pathway are clustered in a single operon that lies just downstream of a -24 TGGC, -12 TTGC nif/ntr-like promoter sequence. Promoters of this class are recognized by a minor form of RNA polymerase utilizing sigma 54 (NtrA, RpoN). Primer extension analysis demonstrated that the dmp operon transcript initiates downstream of the -24, -12 promoter. Transposon insertion mutants, specifically defective in the regulation of the dmp operon, were isolated, and complementation of a phenol-utilization regulatory mutant was used to identify the regulatory locus, dmpR. The 67-kDa dmpR gene product alone was shown to be sufficient for activation of transcription from the dmp operon promoter. Nucleotide sequence determination revealed that DmpR belongs to the NtrC family of transcriptional activators that regulate transcription from -24, -12 promoters. The deduced amino acid sequence of DmpR has high homology (40 to 67% identity) with the central and carboxy-terminal regions of these activators, which are believed to be involved in the interaction with the sigma 54 RNA polymerase and in DNA binding, respectively. The amino-terminal region of DmpR was found to share 64% identity with the amino-terminal region of XylR, which is also a member of this family of activators. This region has been implicated in effector recognition of aromatic compounds that is required for the regulatory activity of XylR.


Phylogeny of cyanobacterial nifH genes: evolutionary implications and potential applications to natural assemblages.
DNA sequences of a fragment of nifH from diverse cyanobacteria were amplified, cloned and sequenced to determine the evolutionary relationship of nitrogenase within the cyanobacteria as a group, and to provide a basis for the identification of uncultivated strains of cyanobacteria in the environment. Analysis of 30 nitrogenase DNA and deduced amino acid sequences from cyanobacteria representing five major taxonomic subdivisions showed great variation in phylogenetic distances between the sequences. Sequences from heterocystous cyanobacteria formed a coherent cluster, in which branching forms did not form a clade distinct from the non-branching forms. Nitrogenase sequences from the unicellular cyanobacteria Gloeothece and Synechococcus sp. RF-1 formed a cluster, as did sequences from the genera Xenococcus and Myxosarcina. The nifH sequences of filamentous nonheterocystous cyanobacteria were not closely related to each other, forming deep branches with respect to the heterocystous cyanobacterial nifH sequences. The phylogeny of nifH based on amino acid sequences was consistent with taxonomic relationships among the strains; for example, a sequence obtained form a natural assemblage believed to be dominated by 'Lyngbya' clustered with nifH from Lyngbya lagerheimii. Results also indicate that the phylogeny of nifH among the cyanobacteria is largely consistent with the phylogeny of 16S rRNA, and furthermore that the nifH sequence can be used to identify uncultivated strains of nitrogen-fixing cyanobacteria.


Neurostimulation for neck pain and headache.
Patients with medically refractory headache disorders are a rare and challenging-to-treat group. The introduction of peripheral neurostimulation (PNS) has offered a new avenue of treatment for patients who are appropriate surgical candidates. The utility of PNS for headache management is actively debated. Preliminary reports suggested that 60-80% of patients with chronic headache who have failed maximum medical therapy respond to PNS. However, complications rates for PNS are high. Recent publication of 2 large randomized clinical trials with conflicting results has underscored the need for further research and careful patient counseling. In this review, we summarize the current evidence for PNS in treatment of chronic migraine, trigeminal autonomic cephalagias and occipital neuralgia, and other secondary headache disorders.


Molecular and expression analysis of the Rhizobium meliloti phosphoenolpyruvate carboxykinase (pckA) gene.
The pckA gene of Rhizobium meliloti, encoding phosphoenolpyruvate carboxykinase, was isolated from a genomic cosmid library by complementation of the succinate growth phenotype of a Pck- mutant. The gene region was mapped by subcloning and Tn5 insertion mutagenesis. The DNA sequence for a 2-kb region containing the structural gene and its promoter was determined. The pckA gene encodes as 536-amino-acid protein that shows homology with other ATP-dependent Pck enzymes. The promoter was identified following primer extension analysis and is similar to sigma 70-like promoters. Expression analysis with a pckA::lacZ gene fusion indicated that the pckA gene was strongly induced at the onset of stationary phase in complex medium. When defined carbon sources were tested, the expression level of the pckA gene was found to be high when cells were grown in minimal media with succinate or arabinose as the sole carbon source but almost absent when glucose, sucrose, or glycerol was the sole carbon source. Glucose and sucrose were not found to strongly repress pckA induction by succinate.


Alcohol-preferring P rats emit spontaneous 22-28 kHz ultrasonic vocalizations that are altered by acute and chronic alcohol experience.
Emotional states are often thought to drive excessive alcohol intake and influence the development of alcohol use disorders. To gain insight into affective properties associated with excessive alcohol intake, we utilized ultrasonic vocalization (USV) detection and analyses to characterize the emotional phenotype of selectively bred alcohol-preferring (P) rats; an established animal model of excessive alcohol intake. USVs emitted by rodents have been convincingly associated with positive (50-55 kHz frequency-modulated [FM]) and negative (22-28 kHz) affective states. Therefore, we hypothesized that 50-55 and 22-28 kHz USV emission patterns in P rats would reveal a unique emotional phenotype sensitive to alcohol experience. 50-55 kHz FM and 22-28 kHz USVs elicited from male P rats were assessed during access to water, 15 and 30% EtOH (v/v). Ethanol (EtOH; n = 12) or water only (Control; n = 4) across 8 weeks of daily drinking-in-the-dark (DID) sessions. Spontaneous 22-28 kHz USVs are emitted by alcohol-naïve P rats and are enhanced by alcohol experience. During DID sessions when alcohol was not available (e.g., "EtOH OFF" intervals), significantly more 22-28 kHz than 50-55 kHz USVs were elicited, while significantly more 50-55 kHz FM than 22-28 kHz USVs were emitted when alcohol was available (e.g., "EtOH ON" intervals). In addition, USV acoustic property analyses revealed chronic effects of alcohol experience on 22-28 kHz USV mean frequency, indicative of lasting alcohol-mediated alterations to neural substrates underlying emotional response. Our findings demonstrate that acute and chronic effects of alcohol exposure are reflected in changes in 22-28 and 50-55 kHz FM USV counts and acoustic patterns. These data support the notion that initiation and maintenance of alcohol intake in P rats may be due to a unique, alcohol-responsive emotional phenotype and further suggest that spontaneous 22-28 kHz USVs serve as behavioral markers for excessive drinking vulnerability.


Structure of a human synaptic GABAA receptor.
Fast inhibitory neurotransmission in the brain is principally mediated by the neurotransmitter GABA (γ-aminobutyric acid) and its synaptic target, the type A GABA receptor (GABAA receptor). Dysfunction of this receptor results in neurological disorders and mental illnesses including epilepsy, anxiety and insomnia. The GABAA receptor is also a prolific target for therapeutic, illicit and recreational drugs, including benzodiazepines, barbiturates, anaesthetics and ethanol. Here we present high-resolution cryo-electron microscopy structures of the human α1β2γ2 GABAA receptor, the predominant isoform in the adult brain, in complex with GABA and the benzodiazepine site antagonist flumazenil, the first-line clinical treatment for benzodiazepine overdose. The receptor architecture reveals unique heteromeric interactions for this important class of inhibitory neurotransmitter receptor. This work provides a template for understanding receptor modulation by GABA and benzodiazepines, and will assist rational approaches to therapeutic targeting of this receptor for neurological disorders and mental illness.


Determinants of black-white differences in the risk of cerebral infarction. The National Health and Nutrition Examination Survey Epidemiologic Follow-up Study.
To determine whether blacks in the First National Health and Nutrition Examination Survey Epidemiologic Follow-up Study remained at increased risk for cerebral infarction after adjusting for stroke risk factors and sociodemographic factors. A cohort study involving 8203 whites and 1362 blacks who participated in the First National Health and Nutrition Examination Survey Epidemiologic Follow-up Study. During the 13-year follow-up, 538 and 122 cerebral infarctions occurred in whites and blacks, respectively. The black-white risk for cerebral infarction varied by age (P = .007 for race-age interaction). Compared with whites of the same age, blacks aged 35 to 44 years were at significantly increased risk for cerebral infarction (relative risk, 2.62; 95% confidence interval, 1.23 to 5.57), while older blacks, those older than 64 years, were not at increased risk (relative risk, 1.14; 95% confidence interval, 0.90 to 1.46). The relative risk for cerebral infarction decreased to 2.07 (95% confidence interval, 0.97 to 4.42) in younger blacks and 0.82 (95% confidence interval, 0.29 to 2.33) in older blacks after adjustment for age, sex, education, history of heart disease, diabetes, systolic blood pressure, treatment for hypertension, Quetelet index, and serum hemoglobin and magnesium levels. These results indicate that much of the increased risk for cerebral infarction experienced by blacks can be explained by their higher prevalence of stroke risk factors, especially diabetes, hypertension, and lower educational attainment. Younger blacks, however, may still be at increased risk after adjusting for stroke risk factors.


Three inositol dehydrogenases involved in utilization and interconversion of inositol stereoisomers in a thermophile, Geobacillus kaustophilus HTA426.
Geobacillus kaustophilus HTA426, a thermophilic Bacillus-related species, utilizes some inositol stereoisomers, including myo-, d-chiro- and scyllo-inositols (MI, DCI and SI), as sole carbon sources. Within its genome are three paralogous genes that possibly encode inositol dehydrogenase. These genes are located in tandem within a large gene cluster containing an almost complete set of iol genes homologous to genes involved in inositol catabolism in Bacillus subtilis. Each of the three plausible inositol dehydrogenases was purified as a His(6)-tag fusion. The enzymes exhibited thermophilic activity, each with its own characteristic specificity for the inositol stereoisomers and cofactors. Northern blot and primer extension analyses revealed that the three enzymes were encoded by the same 5 kb polycistronic transcript and were induced simultaneously in the presence of MI. HTA426 was subjected to ethyl methanesulfonate (EMS) mutagenesis to isolate a mutant strain, PS8, which was not able to utilize MI. In PS8, inositol dehydrogenase activity was abolished along with the 5 kb transcript, suggesting that any of the three enzymes supports MI-dependent growth. Analysis of metabolites in HTA426 cells grown in the presence of MI revealed that substantial amounts of DCI and SI appeared intracellularly during the stationary phase, while only MI was present in PS8 cells, suggesting that interconversion of inositol stereoisomers may involve these three enzymes.


An empirical malaria distribution map for West Africa.
The objective of this study was to produce a malaria distribution map that would constitute a useful tool for development and health planners in West Africa. The recently created continental database of malaria survey results (MARA/ARMA 1998) provides the opportunity for producing empirical models and maps of malaria distribution at a regional and eventually at a continental level. This paper reports on the mapping of malaria distribution for sub-Saharan West Africa based on these data. The strategy was to undertake a spatial statistical analysis of malaria parasite prevalence in relation to those potential bio-physical environmental factors involved in the distribution of malaria transmission intensity which are readily available at any map location. The resulting model was then used to predict parasite prevalence for the whole of West Africa. We also produced estimates of the proportion of population of each country in the region exposed to various categories of risk to show the impact that malaria is having on individual countries. The data represent a very large sample of children in West Africa. It constitutes a first attempt to produce a malaria risk map of the West African region, based entirely on malariometric data. We anticipate that it will provide useful additional guidance to control programme managers, and that it can be refined once sufficient additional data become available.


Bacterial Communities Differ among Drosophila melanogaster Populations and Affect Host Resistance against Parasitoids.
In Drosophila, diet is considered a prominent factor shaping the associated bacterial community. However, the host population background (e.g. genotype, geographical origin and founder effects) is a factor that may also exert a significant influence and is often overlooked. To test for population background effects, we characterized the bacterial communities in larvae of six genetically differentiated and geographically distant D. melanogaster lines collected from natural populations across Europe. The diet for these six lines had been identical for ca. 50 generations, thus any differences in the composition of the microbiome originates from the host populations. We also investigated whether induced shifts in the microbiome-in this case by controlled antibiotic administration-alters the hosts' resistance to parasitism. Our data revealed a clear signature of population background on the diversity and composition of D. melanogaster microbiome that differed across lines, even after hosts had been maintained at the same diet and laboratory conditions for over 4 years. In particular, the number of bacterial OTUs per line ranged from 8 to 39 OTUs. Each line harboured 2 to 28 unique OTUs, and OTUs that were highly abundant in some lines were entirely missing in others. Moreover, we found that the response to antibiotic treatment differed among the lines and significantly altered the host resistance to the parasitoid Asobara tabida in one of the six lines. Wolbachia, a widespread intracellular endosymbiont associated with parasitoid resistance, was lacking in this line, suggesting that other components of the Drosophila microbiome caused a change in host resistance. Collectively, our results revealed that lines that originate from different population backgrounds show significant differences in the established Drosophila microbiome, outpacing the long-term effect of diet. Perturbations on these naturally assembled microbiomes to some degree influenced the hosts' resistance against natural parasites.


Surface slip on rotating graphene membrane enables the temporal selectivity that breaks the permeability-selectivity trade-off.
Membrane separation technology is dictated by the permeability-selectivity trade-off rule, because selectivity relies on membrane pore size being smaller than that of hydrated ions. We discovered a previously unknown mechanism that breaks the permeability-selectivity trade-off in using a rotating nanoporous graphene membrane with pores of 2 to 4 nanometers in diameter. The results show that the rotating membrane exhibits almost 100% salt rejection even when the pore size is larger than that of hydrated ions, and the surface slip at the liquid/graphene interface of rotating membrane enables concurrent ultra-selectivity and unprecedented high permeability. A novel concept of "temporal selectivity" is proposed to attribute the unconventional selectivity to the time difference between the ion's penetration time through the pore and the bypass time required for ion's sliding across the pore. The newly discovered temporal selectivity overcomes the limitation imposed by pore size and provokes a novel theory in designing high-performance membranes.


Body weight, levodopa pharmacokinetics and dyskinesia in Parkinson's disease.
We conducted a pharmacokinetic study in 164 patients with sporadic Parkinson's disease (PD) to address the relationship between body weight and levodopa pharmacokinetics. Patients underwent an oral acute levodopa test with 250 mg levodopa and pharmacokinetic variables were further assessed. Plasmatic levodopa area under the curve (AUC-l) and body weight were significantly and inversely correlated. Women were significantly lighter and more dyskinetic than men, and had greater AUC-l values. Our data suggest that during long-term treatment, lighter PD patients, especially women, may receive a greater cumulative dosage of levodopa per kilogram of body weight. This could explain gender differences for the development of levodopa-induced peak-dose dyskinesias observed during the course of the disease.


Psychrophilic sulfate-reducing bacteria isolated from permanently cold arctic marine sediments: description of Desulfofrigus oceanense gen. nov., sp. nov., Desulfofrigus fragile sp. nov., Desulfofaba gelida gen. nov., sp. nov., Desulfotalea psychrophila gen. nov., sp. nov. and Desulfotalea arctica sp. nov.
Five psychrophilic, Gram-negative, sulfate-reducing bacteria were isolated from marine sediments off the coast of Svalbard. All isolates grew at the in situ temperature of -1.7 degrees C. In batch cultures, strain PSv29T had the highest growth rate at 7 degrees C, strains ASv26T and LSv54T had the highest growth rate at 10 degrees C, and strains LSv21T and LSv514T had the highest growth rate at 18 degrees C. The new isolates used the most common fermentation products in marine sediments, such as acetate, propionate, butyrate, lactate and hydrogen, but only strain ASv26T was able to oxidize fatty acids completely to CO2. The new strains had growth optima at neutral pH and marine salt concentration, except for LSv54T which grew fastest with 1% NaCl. Sulfite and thiosulfate were used as electron acceptors by strains ASv26T, PSv29T and LSv54T, and all strains except PSv29T grew with Fe3+ (ferric citrate) as electron acceptor. Chemotaxonomy based on cellular fatty acid patterns and menaquinones showed good agreement with the phylogeny based on 16S rRNA sequences. All strains belonged to the delta subclass of Proteobacteria but had at least 9% evolutionary distance from known sulfate reducers. Due to the phylogenetic and phenotypic differences between the new isolates and their closest relatives, establishment of the new genera Desulfotalea gen. nov., Desulfofaba gen. nov. and Desulfofrigus gen. nov. is proposed, with strain ASv26T as the type strain of the type species Desulfofrigus oceanense sp. nov., LSv21T as the type strain of Desulfofrigus fragile sp. nov., PSv29T as the type strain of the type species Desulfofaba gelida sp. nov., LSv54T as the type strain of the type species Desulfotalea psychrophila sp. nov. and LSv514T as the type strain of Desulfotalea arctica sp. nov.


Presumptive diagnosis of severe HIV infection to determine the need for antiretroviral therapy in children less than 18 months of age.
To develop a new algorithm for the presumptive diagnosis of severe disease associated with human immunodeficiency virus (HIV) infection in children less than 18 months of age for the purpose of identifying children who require antiretroviral therapy (ART). A conditional probability model was constructed and non-virologic parameters in various combinations were tested in a hypothetical cohort of 1000 children aged 6 weeks, 6 months and 12 months to assess the sensitivity, specificity, and positive and negative predictive values of these algorithms for identifying children in need of ART. The modelled parameters consisted of clinical criteria, rapid HIV antibody testing and CD4+ T-lymphocyte (CD4) count. In children younger than 18 months, the best-performing screening algorithm, consisting of clinical symptoms plus antibody testing plus CD4 count, showed a sensitivity ranging from 71% to 80% and a specificity ranging from 92% to 99%. Positive and negative predictive values were between 61% and 97% and between 95% and 96%, respectively. In the absence of virologic tests, this alternate algorithm for the presumptive diagnosis of severe HIV disease makes it possible to correctly initiate ART in 91% to 98% of HIV-positive children who are at highest risk of dying. The algorithms presented in this paper have better sensitivity and specificity than clinical parameters, with or without rapid HIV testing, for the presumptive diagnosis of severe disease in HIV-positive children less than 18 months of age. If implemented, they can increase the number of HIV-positive children successfully initiated on ART.


The Start Predicts the Finish: Factors Associated With Antidepressant Nonadherence Among Older Veterans During the Acute and Maintenance Treatment Phases.
To best prevent depression relapse and reduce recurrence, an understanding of the factors associated with continued maintenance treatment is needed. This study compared factors associated with antidepressant nonadherence during the acute (ie, during the first 4 months) and maintenance (ie, during 12 months) treatment phase among older veterans with depression. In this prospective, observational study of 278 older veterans with depression (Patient Health Questionnaire-9 score ≥ 5), patients had been given a new antidepressant prescription between 2008 and 2011. Participants completed initial and follow-up interviews at 4 and 12 months. Medication adherence was assessed by the Brief Medication Questionnaire. A generalized estimating equation was used to determine patient factors associated with nonadherence at each time point. Nearly a third of veterans were nonadherent at 4- and 12-month follow-up. In adjusted analyses, nonadherence was significantly associated with African American race (adjusted odds ratio [AOR] = 2.69, 95% CI, 1.30-5.57; P = .01), being unmarried (AOR = 1.84; 95% CI, 1.16-2.92; P = .049), greater medical comorbidity (AOR = 1.30; 95% CI, 1.13-1.49; P < .001), functional impairment (AOR = 1.34; 95% CI, 1.10-1.63; P = .01), and self-reported side effects (AOR = 2.48; 95% CI, 1.57-3.94; P < .001) at both 4 and 12 months. Depression or anxiety severity did not predict antidepressant adherence at either time point. Rates of and factors associated with antidepressant nonadherence were similar at 4 and 12 months. Further work is needed to develop tailored treatment programs to engage older veterans at higher risk of nonadherence in the early treatment period, which may ultimately help to both achieve remission and reduce relapse and recurrence.


Knowledge and perceived competence among nurses caring for the dying in long-term care homes.
The quality of care provided to dying long-term care (LTC) residents is often inadequate, which may be due to the lack of formal training that LTC staff receive in palliative care (PC). The present cross-sectional study assessed PC knowledge and self-efficacy in ability to provide PC in a sample of registered nurses working in LTC homes. A survey was conducted in four LTC homes in October 2009 to June 2010. Nursing staff knowledge of PC was evaluated using the Palliative Care Quiz for Nurses (PCQN). The Self-Efficacy in End-of-Life Care Survey (S-EOLC) was used to measure nursing staff confidence in their ability to provide PC. Close to 60% of the nursing staff participated (69 of 119). The participants did not score highly on the PCQN: the average correct score ranged from 52.50% to 63.41% across the homes. There were no significant differences between the homes for the mean number of correct responses on the PCQN (P=0.329) or mean scores for the three S-EOLC subscales. Rank ordering of the percentage of correct PCQN answers by item and LTC home demonstrated that similar misconceptions were held across homes. Despite their confidence in PC practice, the participants' PC knowledge gap reveals a need for PC training for staff working in LTC homes. The PC education and training provided should both include a gerontological perspective and address the expertise and knowledge already held by staff.


Is nonoperative management of adhesive intestinal obstruction applicable to children in a resource-poor country?
Nonoperative management of adhesive intestinal obstruction gives good results in adults but there are scant studies on its outcome in children. This study reports outcomes and experiences with nonoperative and operative management of adhesive intestinal obstruction in children in a resource-poor country. This is a retrospective analysis of records of children who were managed with adhesive intestinal obstruction at the University of Benin Teaching Hospital between January 2002 and December 2008. Adhesive intestinal obstruction accounted for 21 (8.8%) of 238 children managed with intestinal obstruction. They were aged between 7 weeks and 16 years (mean 3 +/- 6.4 years), comprising 13 males and eight females (ratio 1.6:1). Prior laparotomy for gangrenous/perforated intussusception (seven, 33.3%), perforated appendix (five, 23.8%), perforated volvulus (three, 14.3%), penetrating abdominal trauma (two, 9.5%) and perforated typhoid (two, 9.5%) were major aetiologies. Adhesive obstruction occurred between 6 weeks and 7 years after the index laparotomies. All the 21 children had initial nonoperative management without success, owing to lack of total parenteral nutrition and monitoring facilities. Outcomes of open adhesiolysis performed between 26 and 48 h in six (28.6%) children due to poor response to nonoperative management, 11-13 days in 12 (57.1%) who responded minimally and 2-5 weeks in three (14.3%) who had relapse of symptoms were encouraging. Exploration of the 21 adhesive obstructions confirmed small bowel obstruction due to solitary bands (two, 9.5%), multiple bands/adhesions (13, 61.9%) and encasement, including one bowel gangrene (six, 28.6%). Postoperatively, the only child who had recurrence during 1-6 years of follow-up did well after a repeat adhesiolysis. Nonoperative management was unsuccessful in this setting. Open adhesiolysis may be adopted in children to prevent avoidable morbidities and mortalities in settings with limited resources.


Enhanced output power of GaN-based LEDs with embedded AlGaN pyramidal shells.
In this article, the characteristics of GaN-based LEDs grown on Ar-implanted GaN templates to form inverted Al0.27Ga0.83N pyramidal shells beneath an active layer were investigated. GaN-based epitaxial layers grown on the selective Ar-implanted regions had lower growth rates compared with those grown on the implantation-free regions. This resulted in selective growth, and formation of V-shaped concaves in the epitaxial layers. Accordingly, the inverted Al0.27Ga0.83N pyramidal shells were formed after the Al0.27Ga0.83N and GaN layers were subsequently grown on the V-shaped concaves. The experimental results indicate that the light-output power of LEDs with inverted AlGaN pyramidal shells was higher than those of conventional LEDs. With a 20 mA current injection, the output power was enhanced by 10% when the LEDs were embedded with inverted Al0.27Ga0.83N pyramidal shells. The enhancement in output power was primarily due to the light scattering at the Al0.27Ga0.83N/GaN interface, which leads to a higher escape probability for the photons, that is, light-extraction efficiency. Based on the ray tracing simulation, the output power of LEDs grown on Ar-implanted GaN templates can be enhanced by over 20% compared with the LEDs without the embedded AlGaN pyramidal shells, if the AlGaN layers were replaced by Al0.5Ga0.5N layers.


Correlation between impedance cardiography and B-type natriuretic peptide levels in dyspneic patients.
Both serum measurements of B-type natriuretic peptide (BNP) and impedance cardiography (ICG) are used to evaluate patients for congestive heart failure (CHF) in the Emergency Department (ED), but the agreement between the data obtained by these two methods remains uncertain. We sought to measure the correlation between BNP levels and ICG measurements in patients presenting to the ED with dyspnea, and compare the diagnostic accuracy of each method in diagnosing CHF. We performed a prospective observational study of a convenience sample of patients presenting to the ED with dyspnea and being evaluated for CHF as a cause of their symptoms. An ICG measurement was obtained on each patient and the correlation between BNP level and the ICG parameters cardiac index (CI), systemic vascular resistance (SVR), and thoracic fluid content (TFC) was measured. To further quantify the diagnostic accuracy of ICG and BNP, we then constructed receiver operating characteristic curves based on discharge diagnosis of CHF and compared the area under the curve (AUC) of each test. Out of 54 patients enrolled, correlation was poor between BNP and CI (Spearman's ρ = -.07, p = 0.64) and between BNP and SVR (Spearman's ρ = -.10, p = 0.46), but moderate between BNP and TFC (Spearman's ρ = .32, p = 0.02); a wide degree of scatter was seen in all correlations. BNP levels showed the best diagnostic accuracy for a discharge diagnosis of CHF, with an AUC of .77 (95% confidence interval .59-.95), whereas CI had an AUC of .72 (95% confidence interval .55-.88). We found limited correlation between BNP levels and ICG parameters, suggesting that, in our population, the two tests may not consistently give similar information. BNP level and CI both provided only fair diagnostic accuracy for discharge diagnosis of CHF.


Online Pro-Tobacco Marketing Exposure Is Associated With Dual Tobacco Product Use Among Underage US Students.
To understand the effect of pro-tobacco marketing on electronic cigarette and combustible cigarette dual use among US middle and high school students under 18 years of age. Data were derived from the 2018 National Youth Tobacco Survey, an annual self-administered school-based cross-sectional survey. The survey was administered in public and private middle and high schools across the United States. The probability sample size was 15 238 middle and high school students with complete responses who were under 18 years of age during the study period. The study measured self-reported exposure to online combustible and electronic cigarette advertisements, dual use of combustible and electronic cigarettes during the past 30 days, exposure to the Real Cost antitobacco campaign advertisements, and other sociodemographic factors (eg, race/ethnicity, gender, and grade). Logistic regressions were used to measure pro-tobacco marketing exposure and dual use as a function of pro-tobacco marketing exposure. Descriptive analyses show that 59.0% of respondents were exposed to pro-tobacco online marketing, and 2.9% were dual users. Dual users (odds ratio [OR] = 1.73) and high school students (OR = 1.43) were more likely to report exposure to online pro-tobacco marketing. Findings indicate that a gap in electronic cigarette pro-tobacco marketing regulatory oversight may exist. Further policy action may be warranted to protect the public health of minors and other vulnerable populations who are most susceptible to pro-tobacco marketing.


Explaining Usutu virus dynamics in Austria: model development and calibration.
Usutu virus (USUV), a flavivirus of the Japanese encephalitis virus complex, was for the first time detected outside Africa in the region around Vienna (Austria) in 2001 by Weissenböck et al. [Weissenböck, H., Kolodziejek, J., Url, A., Lussy, H., Rebel-Bauder, B., Nowotny, N., 2002. Emergence of Usutu virus, an African mosquito-borne flavivirus of the Japanese encephalitis virus group, central Europe. Emerg. Infect. Dis. 8, 652-656]. USUV is an arthropod-borne virus (arbovirus) circulating between arthropod vectors (mainly mosquitoes of the Culex pipiens complex) and avian amplification hosts. Infections of mammalian hosts or humans, as observed for the related West Nile virus (WNV), are rare. However, USUV infection leads to a high mortality in birds, especially blackbirds (Turdus merula), and has similar dynamics with the WNV in North America, which, amongst others, caused mortality in American robins (Turdus migratorius). We hypothesized that the transmission of USUV is determined by an interaction of developing proportion of the avian hosts immune and climatic factors affecting the mosquito population. This mechanism is implemented into the present model that simulates the seasonal cycles of mosquito and bird populations as well as USUV cross-infections. Observed monthly climate data are specified for the temperature-dependent development rates of the mosquitoes as well as the temperature-dependent extrinsic-incubation period. Our model reproduced the observed number of dead birds in Austria between 2001 and 2005, including the peaks in the relevant years. The high number of USUV cases in 2003 seems to be a response to the early beginning of the extraordinary hot summer in that year. The predictions indicate that >70% of the bird population acquired immunity, but also that the percentage would drop rapidly within only a couple of years. We estimated annually averaged basic reproduction numbers between R (0)=0.54 (2004) and 1.35 (2003). Finally, extrapolation from our model suggests that only 0.2% of the blackbirds killed by USUV were detected by the Austrian USUV monitoring program [Chvala, S., Bakonyi, T., Bukovsky, C., Meister, T., Brugger, K., Rubel, F., Nowotny, N., Weissenböck, H., 2007. Monitoring of Usutu virus activity and spread by using dead bird surveillance in Austria, 2003-2005. Vet. Microbiol. 122, 237-245]. These results suggest that the model presented is able to quantitatively describe the process of USUV dynamics.


Protein microarrays based on polymer brushes prepared via surface-initiated atom transfer radical polymerization.
Polymer brushes represent an interesting platform for the development of high-capacity protein binding surfaces. Whereas the protein binding properties of polymer brushes have been investigated before, this manuscript evaluates the feasibility of poly(glycidyl methacrylate) (PGMA) and PGMA-co-poly(2-(diethylamino)ethyl methacrylate) (PGMA-co-PDEAEMA) (co)polymer brushes grown via surface-initiated atom transfer radical polymerization (SI-ATRP) as protein reactive substrates in a commercially available microarray system using tantalum-pentoxide-coated optical waveguide-based chips. The performance of the polymer-brush-based protein microarray chips is assessed using commercially available dodecylphosphate (DDP)-modified chips as the benchmark. In contrast to the 2D planar, DDP-coated chips, the polymer-brush-covered chips represent a 3D sampling volume. This was reflected in the results of protein immobilization studies, which indicated that the polymer-brush-based coatings had a higher protein binding capacity as compared to the reference substrates. The protein binding capacity of the polymer-brush-based coatings was found to increase with increasing brush thickness and could also be enhanced by copolymerization of 2-(diethylamino)ethyl methacrylate (DEAEMA), which catalyzes epoxide ring-opening of the glycidyl methacrylate (GMA) units. The performance of the polymer-brush-based microarray chips was evaluated in two proof-of-concept microarray experiments, which involved the detection of biotin-streptavidin binding as well as a model TNFα reverse assay. These experiments revealed that the use of polymer-brush-modified microarray chips resulted not only in the highest absolute fluorescence readouts, reflecting the 3D nature and enhanced sampling volume provided by the brush coating, but also in significantly enhanced signal-to-noise ratios. These characteristics make the proposed polymer brushes an attractive alternative to commercially available, 2D microarray surface coatings.


Familial and "on-the-street" risk factors associated with alcohol use among homeless and runaway adolescents.
This study investigated factors associated with alcohol use among homeless and runaway adolescents, using a risk-amplification model. Homeless and runaway adolescents (N = 536, 60% female) were recruited and interviewed by outreach workers directly on the streets, in shelters and in drop-in centers in four Midwestern states. The average age was 16 years; ages ranged from 12 to 22. Parent alcohol problems were indirectly linked to adolescent drinking through familial abuse and its relationship to deviant peers, time on own and risky subsistence behaviors. Parent alcohol problems also predicted offspring alcohol use through parental rejection and its association with deviant peers and with risky subsistence behaviors. The strongest direct effects on alcohol use were hanging out with antisocial friends and participating in deviant behaviors in order to survive on the street. This study sheds light on the nature of alcohol use in a high-risk population. Family background and "on-the-street" (time on own) factors must be taken into consideration when treating alcohol misuse in street youth. The alternative is a vicious cycle whereby homeless youth may become homeless adults.


Malpractice: provider risk or consumer protection?
The National Practitioner Data Bank (NPDB) began operation in September 1990 as a clearinghouse for adverse action, licensure, and malpractice information in an effort to protect consumers and promote quality in health care. This study analyzed 66,107 and 1291 records of payments made for 50,396 physicians and 1218 nurses, respectively, from 1994 through 1998, to describe characteristics, trends, and risk factors of malpractice payment for physicians and nurses. The median payments, more often settlements paid by insurance companies than judgments in courts of law, were higher for physicians than for nurses. Mean payments were higher for residents than for non-resident physicians; median payments for residents were slightly lower than other physicians when adjusted for number of providers included in the payment. On the state level, correlation analyses suggested a significant positive association between the nurse rate of malpractice payments that were made and median per capita income, number of physicians per 1000 residents, and number of attorneys per 1000 residents; analysis revealed a significant negative association between this rate and the percentage of residents residing in rural areas and the number of nurses per 1000 residents. Although findings suggested that payment trends remained stable, there was great regional variation in the risk of malpractice payment for both physicians and nurses. The physician risk ranged from a low of 0.73% per physician per year in Alabama to a high of 3.7% in Wyoming, and the nurse risk ranged from a low of 0% per nurse per year in Vermont to a high of 0.075% in the District of Columbia. If the quality of health care provided by physicians and nurses does not vary geographically in the United States, then such a great discrepancy seems to challenge the notion that the risk of malpractice litigation consistently promotes the quality of health care.


Prospects for malaria eradication in sub-Saharan Africa.
A characteristic of Plasmodium falciparum infections is the gradual acquisition of clinical immunity resulting from repeated exposures to the parasite. While the molecular basis of protection against clinical malaria remains unresolved, its effects on epidemiological patterns are well recognized. Accumulating epidemiological data constitute a valuable resource that must be intensively explored and interpreted as to effectively inform control planning. Here we apply a mathematical model to clinical data from eight endemic regions in sub-Saharan Africa. The model provides a quantitative framework within which differences in age distribution of clinical disease are assessed in terms of the parameters underlying transmission. The shorter infectious periods estimated for clinical infections induce a regime of bistability of endemic and malaria-free states in regions of mesoendemic transmission. The two epidemiological states are separated by a threshold that provides a convenient measure for intervention design. Scenarios of eradication and resurgence are simulated. In regions that support mesoendemic transmission, intervention success depends critically on reducing prevalence below a threshold which separates endemic and malaria-free regimes.


REV7 counteracts DNA double-strand break resection and affects PARP inhibition.
Error-free repair of DNA double-strand breaks (DSBs) is achieved by homologous recombination (HR), and BRCA1 is an important factor for this repair pathway. In the absence of BRCA1-mediated HR, the administration of PARP inhibitors induces synthetic lethality of tumour cells of patients with breast or ovarian cancers. Despite the benefit of this tailored therapy, drug resistance can occur by HR restoration. Genetic reversion of BRCA1-inactivating mutations can be the underlying mechanism of drug resistance, but this does not explain resistance in all cases. In particular, little is known about BRCA1-independent restoration of HR. Here we show that loss of REV7 (also known as MAD2L2) in mouse and human cell lines re-establishes CTIP-dependent end resection of DSBs in BRCA1-deficient cells, leading to HR restoration and PARP inhibitor resistance, which is reversed by ATM kinase inhibition. REV7 is recruited to DSBs in a manner dependent on the H2AX-MDC1-RNF8-RNF168-53BP1 chromatin pathway, and seems to block HR and promote end joining in addition to its regulatory role in DNA damage tolerance. Finally, we establish that REV7 blocks DSB resection to promote non-homologous end-joining during immunoglobulin class switch recombination. Our results reveal an unexpected crucial function of REV7 downstream of 53BP1 in coordinating pathological DSB repair pathway choices in BRCA1-deficient cells.


What "Tears" Remind Us of: An Investigation of Embodied Cognition and Schizotypal Personality Trait Using Pencil and Teardrop Glasses.
Facial expressions influence our experience and perception of emotions-they not only tell other people what we are feeling but also might tell us what to feel via sensory feedback. We conducted three experiments to investigate the interaction between facial feedback phenomena and different environmental stimuli, by asking participants to remember emotional autobiographical memories. Moreover, we examined how people with schizotypal traits would be affected by their experience of emotional facial simulations. We found that using a directed approach (gripping a pencil with teeth/lips) while remembering a specific autobiographical memory could successfully evoke participants' positive (e.g., happy and excited)/negative (e.g., angry and sad) emotions (i.e., Experiment 1). When using indirective environmental stimuli (e.g., teardrop glasses), the results of our experiments (i.e., Experiments 2 and 3) investigating facial feedback and the effect of teardrop glasses showed that participants who scored low in schizotypy reported little effect from wearing teardrop glasses, while those with high schizotypy reported a much greater effect in both between- and within-subject conditions. The results are discussed from the perspective of sense of ownership, which people with schizophrenia are believed to have deficits in.


Transcription Factors Drive Tet2-Mediated Enhancer Demethylation to Reprogram Cell Fate.
Here, we report DNA methylation and hydroxymethylation dynamics at nucleotide resolution using C/EBPα-enhanced reprogramming of B cells into induced pluripotent cells (iPSCs). We observed successive waves of hydroxymethylation at enhancers, concomitant with a decrease in DNA methylation, suggesting active demethylation. Consistent with this finding, ablation of the DNA demethylase Tet2 almost completely abolishes reprogramming. C/EBPα, Klf4, and Tfcp2l1 each interact with Tet2 and recruit the enzyme to specific DNA sites. During reprogramming, some of these sites maintain high levels of 5hmC, and enhancers and promoters of key pluripotency factors become demethylated as early as 1 day after Yamanaka factor induction. Surprisingly, methylation changes precede chromatin opening in distinct chromatin regions, including Klf4 bound sites, revealing a pioneer factor activity associated with alternation in DNA methylation. Rapid changes in hydroxymethylation similar to those in B cells were also observed during compound-accelerated reprogramming of fibroblasts into iPSCs, highlighting the generality of our observations.


Persistence of repeated sequences that evolve by replication slippage.
The evolution of short repeated sequences by replication slippage under the assumption of selective neutrality is modeled using a linear birth and death process. The equilibrium distribution, the distribution of the life expectancy of a repeated sequence when the process starts from two repeats, the age distribution of repeats, the probability of obtaining two genes with i and j copies which diverged t generations ago and the conditional variance of copy number given the repeat number is more than one are computed. The distributions of life expectancy and age are shown to have long tails. Also the statistic which estimates the conditional variance is shown to have a large coefficient of variation. Using these theoretical results, we develop an approximate test of our model and analyze persistent repeated sequences found in the primate beta-globin gene region and Oenothera chloroplast DNA which are polymorphic within species. We found one sequence in Oenothera chloroplast DNA which does not fit to our neutral model.


The protective effects of acetyl L-carnitine on testis gonadotoxicity induced by Cisplatin in rats.
Cisplatin, an effective antineoplastic agent, damages normal cells in a manner related to chemotherapy. Acetyl L-carnitine protects cells against mitochondrial and nuclear damage induced by chemotherapy. Animal experiment. The aim of this study was to examine the protective effects of acetyl L-carnitine on cisplatin-induced gonadotoxicity in testicular structures. Twenty-four male Wistar albino rats were divided into four Groups (n=6): Group 1 (control) was administered saline; Group 2 was administered acetyl L-carnitine; Group 3 was administered cisplatin; and Group 4 was pre-treated with acetyl L-carnitine before cisplatin administration. After 72hr of treatment with cisplatin, the rats were sacrificed, and the testicular tissues were removed. Morphometric, histomorphologic and immunohistochemical analyses were conducted. At the end of the experiment, Group 3 was characterised by statistically significant weight loss, a degenerative appearance of the seminiferous tubules in the peripheral region, separation of spermatogenic cell series from the tubular wall, cellular debris in the lumen and central interstitial oedema. Sperm morphology appeared to be abnormal. Tubular diameter and wall thickness decreased, and the number of TUNEL- and active caspase-positive cells increased compared with the other Groups. The histological findings in Group 4 were better than those in Group 3. It was concluded that the prophylactic use of acetyl L-carnitine protects against cisplatin-induced testicular tissue damage.


Efficacy of Biodentine and Mineral Trioxide Aggregate in Primary Molar Pulpotomies-A Systematic Review and Meta-Analysis With Trial Sequential Analysis of Randomized Clinical Trials.
Pulpotomy is the favored treatment for pulp exposure in carious primary teeth. This review aimed to compare the success rates of biodentine (BD) and mineral trioxide aggregate (MTA) pulpotomies in primary molars using meta-analysis (MA) and trial sequential analysis (TSA) and also to assess the quality of the results by Grading of Recommendations, Assessment, Development and Evaluation (GRADE). PubMed, EBSCOhost, and Scopus databases were searched. Additional searching was performed in clinical trial registry, reference lists of systematic reviews, and textbooks. Randomized clinical trials (RCTs) published in the English language through October 2017 comparing the success of pulpotomies in vital primary molars with a follow-up of at least 6 months were selected. Study selection, data extraction, and risk of bias assessment were performed. MA by random effects model, TSA, and GRADE were performed. Eight RCTs (n = 474) were included. Two RCTs had low risk of bias. No significant difference was observed between MTA and BD in clinical success at 6 months (risk ratio [RR], 1.00; 95% confidence interval [95% CI], 0.97-1.02; I2 = 0%), 12 months (RR, 1.00; 95% CI, 0.96-1.05; I2 = 0%), and 18 months (RR, 1.00; 95% CI, 0.93-1.08; I2 = 0%). No difference was observed in radiographic success at follow-up of 6 months (RR, 0.99; 95% CI, 0.96-1.02; I2 = 0%), 12 months (RR, 1.02; 95% CI, 0.47-2.21; I2 = 0%), and 18 months (RR, 1.02; 95% CI, 0.91-1.15; I2 = 0%). TSA indicated lack of firm evidence for the results of the meta-analytic outcomes on clinical and radiographic success. GRADE assessed the evidence from the MA comparing the effect of MTA and BD in pulpotomy to be of low quality. BD and MTA have similar clinical and radiographic success rates based on limited and low-quality evidence. Future high-quality RCTs between MTA and BD is required to confirm the evidence.


Metabolism of 2,3-dihydroxypropane-1-sulfonate by marine bacteria.
Both enantiomers of the sulfoquinovose breakdown product 2,3-dihydroxypropane-1-sulfonate, an important sulfur metabolite produced by marine algae, were synthesised in a 34S-labelled form and used in feeding experiments with marine bacteria. The labelling was efficiently incorporated into the sulfur-containing antibiotic tropodithietic acid and sulfur volatiles by the algal symbiont Phaeobacter inhibens, but not into sulfur volatiles released by marine bacteria associated with crustaceans. The ecological implications and the relevance of these findings for the global sulfur cycle are discussed.


Inherited thrombotic thrombocytopenic purpura in pregnancy.
The primary pathologic reason for thrombotic thrombocytopenic purpura (TTP) lies in the systemic formation of platelet aggregations in association with endothelial cells damage. Endothelial damage is a result of an abnormal synthesis and metabolism of unusually large von Willebrand Factor (ULvWF) multimers. In normal conditions vWF cleaving metalloprotease, known as ADAMTS-13 (A Disintegrin And Metalloproteinase with Thrombospondin type-1 motif, member 13) prevents the ULvWF entrance in the circulation. It already has been proven that thrombotic thrombocytopenic purpura is strongly correlated with severe congenital or acquired deficiency of ADAMTS-13. Congenital ADAMTS-13 deficiency is known as Upshaw-Schulman Syndrome and it accounts for only 2-4% of all TTP cases. It is conditioned by genetic variants of the ADAMTS-13 gene causing reduced ADAMTS-13 synthesis and shows an autosomal recessive type of inheritance. We present an interesting case of a 20 year old patient, primigravida, nulliparous, in 28th gestational week of twin pregnancy with undiagnosed Upshaw-Schulman Syndrome. The patient was transferred from the district hospital to the Tertiary Perinatal Care Center because of thrombocytopenia and suspicion of hemolytic-uremic syndrome. Genetic analysis performed 5 years after patient's death (before 2008 that kind of genetic analysis had not been available in Poland) showed a homozygotic mutation in the ADMTS13 gene (4143insA) which confirmed the diagnosis of Upshaw-Schulman Syndrome. 1. TTP, especially hereditary Upshaw-Schulman Syndrome, is extremely rare and complicates the course of pregnancy. It is usually very sudden and dramatic; 2. Differential diagnosis of this disease is difficult and treatment strategy very burdensome for the patient. For this reason, diagnosis of micro-angiopathic disorders need to be simultaneously based on both clinical symptoms and laboratory findings; 3. Genetic diagnosis that confirms exact recognition of Upshaw-Schulman Syndrome is not commonly available; 4. The described case was a huge diagnostic challenge, and actually the final diagnosis was published until five years after the patient's death. Before 2008, that type of genetic analysis had not been available in Poland; 5. Despite the enormous progress in medical knowledge and experience, the exact diagnosis of TTP, including Upshaw-Schulman Syndrome, of this condition remains very difficult.


Utility transforms of optical fields employing deformable mirror.
Diffraction optical devices of modest degrees of freedom (DOF), such as deformable mirrors, have not been exploited as general-purpose transformers of optical fields. Described in this Letter is a method that guides deformable mirrors to optimal surfaces allowed by the DOF for various desirable outcomes. The method is based on a modal optimization procedure with the help of Walsh functions in controlling the variables of the mirrors, i.e. the actuators. It is shown that a deformable mirror of modest DOF can provide field transformations for arbitrary beam-splitting, formation of ring-shaped beams, and coherent beam combining.


Differentiated melanocyte cell division occurs in vivo and is promoted by mutations in Mitf.
Coordination of cell proliferation and differentiation is crucial for tissue formation, repair and regeneration. Some tissues, such as skin and blood, depend on differentiation of a pluripotent stem cell population, whereas others depend on the division of differentiated cells. In development and in the hair follicle, pigmented melanocytes are derived from undifferentiated precursor cells or stem cells. However, differentiated melanocytes may also have proliferative capacity in animals, and the potential for differentiated melanocyte cell division in development and regeneration remains largely unexplored. Here, we use time-lapse imaging of the developing zebrafish to show that while most melanocytes arise from undifferentiated precursor cells, an unexpected subpopulation of differentiated melanocytes arises by cell division. Depletion of the overall melanocyte population triggers a regeneration phase in which differentiated melanocyte division is significantly enhanced, particularly in young differentiated melanocytes. Additionally, we find reduced levels of Mitf activity using an mitfa temperature-sensitive line results in a dramatic increase in differentiated melanocyte cell division. This supports models that in addition to promoting differentiation, Mitf also promotes withdrawal from the cell cycle. We suggest differentiated cell division is relevant to melanoma progression because the human melanoma mutation MITF(4T)(Δ)(2B) promotes increased and serial differentiated melanocyte division in zebrafish. These results reveal a novel pathway of differentiated melanocyte division in vivo, and that Mitf activity is essential for maintaining cell cycle arrest in differentiated melanocytes.


Investigation of 3D-Printed Polycaprolactone-/Polyvinylpyrrolidone-Based Constructs.
The aim of this study is to evaluate the mechanical and biological performance of cartilage-like constructs produced by 3D printing. During the investigation, poly(ε-caprolactone) (PCL) and polyvinylpyrrolidone (PVP) were used as a matrix polymer and low-molecular-weight chitosan (CS), hyaluronic acid (HA), and alginic acid sodium salt (SA) were integrated separately with the polymer matrix to fabricate the constructs. Thermal, mechanical, morphology, and chemical properties and swelling, degradation, and biocompatibility behaviors were evaluated in detail. With the addition of 3 fillers, the melting temperature of the matrix increased with the addition of fillers, and PCL/3wt.%PVP/1wt.%HA had the highest melting temperature value. Mechanical characterization results demonstrated that the printed PCL/3wt.%PVP/1wt.%CS displayed the highest compressive strength of around 9.51 MPa. The compressive strength difference between the PCL/3wt.%PVP and PCL/3wt.%PVP/1wt.%CS was 5.38 MPa. Biocompatibility properties of the constructs were tested by mitochondrial dehydrogenase activity, and in vitro studies showed that the PCL/3wt.%PVP/1wt.%HA composite construct had more cell viability than the other constructs by making use of the mesenchymal stem cell line.


Physical activity as a preventive measure against overweight, obesity, infections, allergies and cardiovascular disease risk factors in adolescents: AFINOS Study protocol.
Prior studies addressing the impacts of regular physical activity or sedentary habits on the immune system have been conducted in adults and laboratory settings. Thus, it is practically unknown how a healthy active lifestyle could affect low-grade inflammation processes, infections or allergies in young persons. The AFINOS Study was designed to determine the relationship between the regular physical activity levels of adolescents and overweight, infection, and allergies along with the presence of metabolic and immunological biomarkers of a deteriorated health status. A further objective of the AFINOS Study is to assess the health status and lifestyle habits of an adolescent population in an effort to identify any protective factors that could be used as preventive measures, since many chronic diseases and their associated co-morbidities often persist from adolescence into adulthood. This study was conducted as three separate sub-studies in three different populations as follows: (a) Study 1 was performed on a population sample of adolescents; (b) Study 2 on the adolescents' parents; and (c) Study 3 on a subset of the adolescents from Study 1. Study 1 assessed health and lifestyle indicators through a questionnaire administered to a representative sample of adolescents from the Madrid Region (n = 2400) aged 13 to 16 years. In Study 2, the parents of the teenagers participating in Study 1 were required to fill out a questionnaire. Finally in Study 3, body composition, physical activity, health-related physical fitness, and blood measurements were determined in a subset (n = 200) of the individuals included in Study 1. This paper describes the rationale, design, and methodologies used in the AFINOS Study. This multidisciplinary, multicenter study seeks to evaluate several aspects of existing relationships between routine physical activity/sedentary behaviour and several health status markers, specifically those related to the immune system. The results of this cross-sectional study will serve for comparisons with the available data obtained in laboratory settings and in adults. In addition, knowledge regarding the health status and lifestyle habits of Spanish adolescents and their parents will be useful for designing preventive measures.


The molecular pathogenesis of migraine: new developments and opportunities.
Migraine is a prevalent, debilitating and costly disorder with an ongoing unmet medical need. Human genetic studies have provided considerable insights into the molecular underpinnings of this complex brain disorder. Classical linkage studies have revealed the causes of familial hemiplegic migraine, while more recently genome-wide association studies have identified several susceptibility loci for typical migraine. New ways of accessing neurons and other cells directly from patients with migraine through the use of induced pluripotent stem cells offer exciting opportunities to understand the molecular pathogenesis. In conjunction with next generation omics, there are unprecedented opportunities to reveal key molecular players in the disease process and discover new drug targets.


Mechanisms of convergence and extension by cell intercalation.
The cells of many embryonic tissues actively narrow in one dimension (convergence) and lengthen in the perpendicular dimension (extension). Convergence and extension are ubiquitous and important tissue movements in metazoan morphogenesis. In vertebrates, the dorsal axial and paraxial mesodermal tissues, the notochordal and somitic mesoderm, converge and extend. In amphibians as well as a number of other organisms where these movements appear, they occur by mediolateral cell intercalation, the rearrangement of cells along the mediolateral axis to produce an array that is narrower in this axis and longer in the anteroposterior axis. In amphibians, mesodermal cell intercalation is driven by bipolar, mediolaterally directed protrusive activity, which appears to exert traction on adjacent cells and pulls the cells between one another. In addition, the notochordal-somitic boundary functions in convergence and extension by 'capturing' notochordal cells as they contact the boundary, thus elongating the boundary. The prospective neural tissue also actively converges and extends parallel with the mesoderm. In contrast to the mesoderm, cell intercalation in the neural plate normally occurs by monopolar protrusive activity directed medially, towards the midline notoplate-floor-plate region. In contrast, the notoplate-floor-plate region appears to converge and extend by adhering to and being towed by or perhaps migrating on the underlying notochord. Converging and extending mesoderm stiffens by a factor of three or four and exerts up to 0.6 microN force. Therefore, active, force-producing convergent extension, the mechanism of cell intercalation, requires a mechanism to actively pull cells between one another while maintaining a tissue stiffness sufficient to push with a substantial force. Based on the evidence thus far, a cell-cell traction model of intercalation is described. The essential elements of such a morphogenic machine appear to be (i) bipolar, mediolaterally orientated or monopolar, medially directed protrusive activity; (ii) this protrusive activity results in mediolaterally orientated or medially directed traction of cells on one another; (iii) tractive protrusions are confined to the ends of the cells; (iv) a mechanically stable cell cortex over the bulk of the cell body which serves as a movable substratum for the orientated or directed cell traction. The implications of this model for cell adhesion, regulation of cell motility and cell polarity, and cell and tissue biomechanics are discussed.


Constitutively active AKT depletes hematopoietic stem cells and induces leukemia in mice.
Human cancers, including acute myeloid leukemia (AML), commonly display constitutive phosphoinositide 3-kinase (PI3K) AKT signaling. However, the exact role of AKT activation in leukemia and its effects on hematopoietic stem cells (HSCs) are poorly understood. Several members of the PI3K pathway, phosphatase and tensin homolog (Pten), the forkhead box, subgroup O (FOXO) transcription factors, and TSC1, have demonstrated functions in normal and leukemic stem cells but are rarely mutated in leukemia. We developed an activated allele of AKT1 that models increased signaling in normal and leukemic stem cells. In our murine bone marrow transplantation model using a myristoylated AKT1 (myr-AKT), recipients develop myeloproliferative disease, T-cell lymphoma, or AML. Analysis of the HSCs in myr-AKT mice reveals transient expansion and increased cycling, associated with impaired engraftment. myr-AKT-expressing bone marrow cells are unable to form cobblestones in long-term cocultures. Rapamycin, an inhibitor of the mammalian target of rapamycin (mTOR) rescues cobblestone formation in myr-AKT-expressing bone marrow cells and increases the survival of myr-AKT mice. This study demonstrates that enhanced AKT activation is an important mechanism of transformation in AML and that HSCs are highly sensitive to excess AKT/mTOR signaling.


Partial adenosine A1 receptor agonism: a potential new therapeutic strategy for heart failure.
Heart failure (HF) represents a global public health and economic problem associated with unacceptable rates of death, hospitalization, and healthcare expenditure. Despite available therapy, HF carries a prognosis comparable to many forms of cancer with a 5-year survival rate of ~50%. The current treatment paradigm for HF with reduced ejection fraction (EF) centers on blocking maladaptive neurohormonal activation and decreasing cardiac workload with therapies that concurrently lower blood pressure and heart rate. Continued development of hemodynamically active medications for stepwise addition to existing therapies carries the risk of limited tolerability and safety. Moreover, this treatment paradigm has thus far failed for HF with preserved EF. Accordingly, development of hemodynamically neutral HF therapies targeting primary cardiac pathologies must be considered. In this context, a partial adenosine A1 receptor (A1R) agonist holds promise as a potentially hemodynamically neutral therapy for HF that could simultaneous improve cardiomyocyte energetics, calcium homeostasis, cardiac structure and function, and long-term clinical outcomes when added to background therapies. In this review, we describe the physiology and pathophysiology of HF as it relates to adenosine agonism, examine the existing body of evidence and biologic rationale for modulation of adenosine A1R activity, and review the current state of drug development of a partial A1R agonist for the treatment of HF.


The effect of bedding system selected by manual muscle testing on sleep-related cardiovascular functions.
Different types of mattresses affect sleep quality and waking muscle power. Whether manual muscle testing (MMT) predicts the cardiovascular effects of the bedding system was explored using ten healthy young men. For each participant, two bedding systems, one inducing the strongest limb muscle force (strong bedding system) and the other inducing the weakest limb force (weak bedding system), were identified using MMT. Each bedding system, in total five mattresses and eight pillows of different firmness, was used for two continuous weeks at the participant's home in a random and double-blind sequence. A sleep log, a questionnaire, and a polysomnography were used to differentiate the two bedding systems. Heart rate variability and arterial pressure variability analyses showed that the strong bedding system resulted in decreased cardiovascular sympathetic modulation, increased cardiac vagal activity, and increased baroreceptor reflex sensitivity during sleep as compared to the weak bedding system. Different bedding systems have distinct cardiovascular effects during sleep that can be predicted by MMT.


Tracking antigen-experienced effector T cells in vitro and in vivo.
The TCR complex is a multisubunit complex, comprising at least eight transmembrane units. The clonotypic TCR alpha and beta chains are responsible for antigen recognition, whilst the invariant chains of the CD3 complex (delta, epsilon and gamma) and two zeta (zeta) polypeptides couple antigen recognition to downstream signal transduction pathways. TCRzeta (CD247) functions as an amplification module in the TCR signalling cascade and is also essential for the assembly and surface expression of the TCR/CD3 complex. Loss of TCRzeta expression is common in chronic infectious and inflammatory diseases, as well as in cancer. Previous work has indicated that TCRzeta(low)-expressing cells phenotypically resemble antigen-experienced effector T cells. Here, we describe the derivation of a flow cytometry-based TCRzeta expression index for the purpose of more precisely defining TCRzeta expression, in addition to utilising a simple transmigration assay in the demonstration that TCRzeta(dim) T cells have intrinsic migratory properties that may explain their accumulation at sites of inflammation.