Anti-HIV I/II activity and molecular cloning of a novel mannose/sialic acid-binding lectin from rhizome of Polygonatum cyrtonema Hua.
The anti-human immunodeficiency virus (HIV) I/II activity of a mannose and sialic acid binding lectin isolated from rhizomes of Polygonatum cyrtonema Hua was elucidated by comparing its HIV infection inhibitory activity in MT-4 and CEM cells with that of other mannose-binding lectins (MBLs). The anti-HIV activity of Polygonatum cyrtonema Hua lectin (PCL) was 10- to 100-fold more potent than other tested MBLs, but without significant cytotoxicity towards MT-4 or CEM cells. To amplify cDNA of PCL by 3'/5'-rapid amplification of cDNA ends (RACE), the 30 amino acids of N-terminal were determined by sequencing and the degenerate oligonucleotide primers were designed. The full-length cDNA of PCL contained 693 bp with an open reading frame encoding a precursor protein of 160 amino acid residues, consisting of a 28-residue signal peptide, a 22-residue C-terminal cleavage peptide and a 110-residue mature polypeptide which contained three tandemly arranged subdomains with an obvious sequence homology to the monocot MBL. However, only one active mannose-binding site (QDNVY) was found in subdomain I of PCL, that of subdomain II and III changed to HNNVY and PDNVY, respectively. There was no intron in PCL, which was in good agreement with other monocot MBLs. Molecular modeling of PCL indicated that its three-dimensional structure resembles that of the snowdrop agglutinin. By docking, an active sialic acid-binding site was found in PCL. The instabilization of translation initiation region (TIR) in mRNA of PCL benefits its high expression in rhizomes.


Computational neuroimaging strategies for single patient predictions.
Neuroimaging increasingly exploits machine learning techniques in an attempt to achieve clinically relevant single-subject predictions. An alternative to machine learning, which tries to establish predictive links between features of the observed data and clinical variables, is the deployment of computational models for inferring on the (patho)physiological and cognitive mechanisms that generate behavioural and neuroimaging responses. This paper discusses the rationale behind a computational approach to neuroimaging-based single-subject inference, focusing on its potential for characterising disease mechanisms in individual subjects and mapping these characterisations to clinical predictions. Following an overview of two main approaches - Bayesian model selection and generative embedding - which can link computational models to individual predictions, we review how these methods accommodate heterogeneity in psychiatric and neurological spectrum disorders, help avoid erroneous interpretations of neuroimaging data, and establish a link between a mechanistic, model-based approach and the statistical perspectives afforded by machine learning.


A systematic review of the efficacy and safety of vaginal estrogen products for the treatment of genitourinary syndrome of menopause.
We updated a systematic review to evaluate the totality of evidence available for the efficacy and safety of vaginal estrogen products for the treatment of genitourinary syndrome of menopause (GSM) based on published randomized controlled trials. We searched the Cochrane Library, Ovid, PubMed, Medline, Embase, and Clinicaltrials.gov for English-language articles from database inception to June 2018. Our search consolidated 2,086 potential sources to 53 full-text articles that were reviewed and found relevant to our systematic review. We identified 53 studies that met the inclusion criteria that evaluated the efficacy and safety of vaginal estrogen versus placebo or other hormone and nonhormone controls. Compared with placebo, all vaginal estrogens demonstrated superiority in objective endpoints and subjective endpoints of GSM, whereas some trials demonstrated superiority versus placebo in urogenital symptoms. No significant difference was observed between various dosages and dosage forms of vaginal estrogen products. Vaginal estrogen showed superiority over vaginal lubricants and moisturizers for the improvement of objective clinical endpoints of vulvovaginal atrophy but not for subjective endpoints. Unopposed vaginal estrogens seemed safe, although studies were not powered to detect a long-term estrogenic side effect. Estrogen products were found to be clinically effective for the treatment of GSM with doses as low as 4 μg. Vaginal estrogen products seem to be safe with few adverse effects, although there is a lack of long-term controlled clinical trial safety data. This review supports the use of commercially available vaginal estrogen therapies as an effective and safe first-line therapy for the treatment of moderate-to-severe GSM.


In vivo electroporation and non-protein based screening assays to identify antibodies against native protein conformations.
In vivo electroporation has become a gold standard method for DNA immunization. The method assists the DNA entry into cells, results in expression and the display of the native form of antigens to professional cells of the immune system, uses both arms of immune system, has a built-in adjuvant system, is relatively safe, and is cost-effective. However, there are challenges for achieving an optimized reproducible process for eliciting strong humoral responses and for the screening of specific immune responses, in particular, when the aim is to mount humoral responses or to generate monoclonal antibodies via hybridoma technology. Production of monoclonal antibodies demands generation of high numbers of primed B and CD4 T helper cells in lymphoid organs needed for the fusion that traditionally is achieved by a final intravenous antigen injection. The purified antigen is also needed for screening of hundreds of clones obtained upon fusion of splenocytes. Such challenges make DNA vaccination dependent on purified proteins. Here, we have optimized methods for in vivo electroporation, production, and use of cells expressing the antigen and an in-cell Western screening method. These methods resulted in (1) reproducibly mounting robust humoral responses against antigens with different cell localizations, and (2) the ability to screen for antigen eliminating a need for protein/antigen purification. This process includes optimized parameters for in vivo electroporation, the use of transfected cells for final boost, and mild fixation/permeabilization of cells for screening. Using this process, upon two vaccinations via in vivo electroporation (and final boost), monoclonal antibodies against nucleus and cytoplasmic and transmembrane proteins were achieved.


A novel miR-1291-ERRα-CPT1C axis modulates tumor cell proliferation, metabolism and tumorigenesis.
Rationale: MicroRNAs are known to influence the development of a variety of cancers. Previous studies revealed that miR-1291 has antiproliferative functions in cancer cells. Carnitine palmitoyltransferase 1C (CPT1C) has a vital role in mitochondrial energy metabolism and modulation of cancer cell proliferation. Since both miR-1291 and CPT1C regulate tumor cell metabolism and cancer progression, we hypothesized that they might be regulated synergistically. Methods: A series of cell phenotype indicators, such as BrdU, colony formation, cell cycle, ATP production, ROS accumulation and cell ability to resist metabolic stress, were performed to clarify the effects of miR-1291 and ERRα expression on tumor cell proliferation and metabolism. A xenograft tumor model was used to evaluate cell tumorigenesis. Meta-analysis and bioinformatic prediction were applied in the search for the bridge-link between miR-1291 and CPT1C. RT-qPCR, western-blot and IHC analysis were used for the detection of mRNA and protein expression. Luciferase assays and ChIP assays were conducted for in-depth mechanism studies. Results: The expression of miR-1291 inhibited growth and tumorigenesis as a result of modulation of metabolism. CPT1C expression was indirectly and negatively correlated with miR-1291 levels. ESRRA was identified as a prominent differentially expressed gene in both breast and pancreatic cancer samples, and estrogen-related receptor α (ERRα) was found to link miR-1291 and CPT1C. MiR-1291 targeted ERRα and CPT1C was identified as a newly described ERRα target gene. Moreover, ERRα was found to influence cancer cell metabolism and proliferation, consistent with the cellular changes caused by miR-1291. Conclusion: This study demonstrated the existence and mechanism of action of a novel miR-1291-ERRα-CPT1C cancer metabolism axis that may provide new insights and strategies for the development of miRNA-based therapies for malignant cancers.


Diversity of oligosaccharide structures on the envelope glycoprotein gp 120 of human immunodeficiency virus 1 from the lymphoblastoid cell line H9. Presence of complex-type oligosaccharides with bisecting N-acetylglucosamine residues.
The N-linked oligosaccharide structures on the envelope glycoprotein gp120 of human immunodeficiency virus 1 derived from chronically infected lymphoblastoid (H9) cells have been investigated by enzymatic microsequencing after release from protein by hydrazinolysis, labeling with NaB3H4, and chromatography on adsorbent columns of Phaseolus vulgaris erythrophytohemagglutinin and Ricinus communis agglutinin (Mr 120,000) and on Bio-Gel P-4. A substantially greater diversity of oligosaccharide structures was detected than among those released by hydrazinolysis from recombinant gp120 produced in Chinese hamster ovary cells and investigated by similar procedures (Mizuochi, T., Spellman, M.W., Larkin, M., Solomon, J., Basa, L.J., and Feizi, T. (1988) Biochem J. 254, 599-603) and among those released by endoglycosidases from virus-derived gp120 isolated from infected H9 cells after metabolic labeling with D-[2-3H]mannose or D-[6-3H]glucosamine (Geyer, H., Holschbach, L., Hunsmann, G., and Schneider, J. (1988) J. Biol. Chem. 263, 11760-11767). In this study, 16% of the oligosaccharides were identified as complex-type bi-, tri-, and tetraantennary sialo-oligosaccharides with bisecting N-acetylglucosamine residues. Such structures were lacking on recombinant gp120 and could not be detected on the metabolically labeled, virus-derived glycoprotein. As in the earlier investigations, complex-type chains lacking bisecting N-acetylglucosamine residues, hybrid-type chains, and a series of high mannose-type structures with 5-9 mannose residues were identified. In addition, an array of complex-type chains having one or more outer chains with beta-galactosyl residues were detected in this study, but with additional substitutions that require further investigation. The number of potential N-glycosylation sites on gp120 is on the order of 20, but the oligosaccharide structures are far more numerous. Thus, the salient conclusion from this and earlier investigations is that alternative structures occur on at least some of the glycosylation sites and that numerous glycosylation variants of this glycoprotein are produced even within a single cell line. Since the glycosylation is the product of host cell glycosyltransferases, an even greater number of glycosylation variants of gp120 are predicted to arise from the heterogeneous cell populations harboring the virus in in vivo infection.


DH166, a beta-carboline derivative, inhibits the kinase activity of PLK1.
A better way to treat complex diseases such as cancer is to aim for several targets at once. Beta-carboline derivatives have been shown to have anticancer activity, but these compounds may target several enzymes required for cell division. Polo-like kinases (PLKs) are well conserved serine/threonine kinases and PLK1 plays multiple roles in cell proliferation. Thus, PLK1 is one of the attractive mitotic targets for anticancer drugs. We found that DH166, a beta-carboline derivative, inhibits the growth of cdc5-2 temperature-sensitive mutant more profoundly than wild-type yeast cells. Because Cdc5 is the human PLK1 homologue in budding yeast, this observation indicates that DH166 might be a PLK1 inhibitor. Indeed, DH166 inhibits the kinase activity of purified PLK1 at low micromolar concentration in an ATP-competitive manner, which is consistent with the docking result based on the crystal structure of PLK1. In addition, DH166 blocks cancer cell proliferation, causes a mitotic arrest, increases cyclin B1 accumulation, induces aberrant mitotic spindles and apoptosis, presumably due to the downregulation of PLK1. Although beta-carboline derivatives have been demonstrated to show antitumor activities through multiple mechanisms, our data indicate for the first time that their cytotoxicity to tumor cells might be attributable to the inhibition of PLK1 as well.


Depletion of potassium and sodium in mantles of Mars, Moon and Vesta by core formation.
The depletions of potassium (K) and sodium (Na) in samples from planetary interiors have long been considered as primary evidence for their volatile behavior during planetary formation processes. Here, we use high-pressure experiments combined with laser ablation analyses to measure the sulfide-silicate and metal-silicate partitioning of K and Na at high pressure (P) - temperature (T) and find that their partitioning into metal strongly increases with temperature. Results indicate that the observed Vestan and Martian mantle K and Na depletions can reflect sequestration into their sulfur-rich cores in addition to their volatility during formation of Mars and Vesta. This suggests that alkali depletions are not affected solely by incomplete condensation or partial volatilization during planetary formation and differentiation, but additionally or even primarily reflect the thermal and chemical conditions during core formation. Core sequestration is also significant for the Moon, but lunar mantle depletions of K and Na cannot be reconciled by core formation only. This supports the hypothesis that measured isotopic fractionations of K in lunar samples represent incomplete condensation or extensive volatile loss during the Moon-forming giant impact.


Messages to Motivate Human Papillomavirus Vaccination: National Studies of Parents and Physicians.
Physician communication about human papillomavirus (HPV) vaccine is a key determinant of uptake. To support physician communication, we sought to identify messages that would motivate HPV vaccination. From 2014 to 2015, we surveyed national samples of parents of adolescents ages 11 to 17 (n = 1,504) and primary care physicians (n = 776). Parents read motivational messages, selected from nine longer messages developed by the Centers for Disease Control and Prevention and six brief messages developed by the study team. Parents indicated whether each message would persuade them to get HPV vaccine for their adolescents. Physicians read the brief messages and indicated whether they would use them to persuade parents to get HPV vaccine for 11- to 12-year-old children. The highest proportion of parents (65%) and physicians (69%) found this brief message to be persuasive: "I strongly believe in the importance of this cancer-preventing vaccine for [child's name]." Parents disinclined to vaccinate were most receptive to messages with information about HPV infection being common, cancers caused by HPV, and HPV vaccine effectiveness. Parents' endorsement did not vary by race/ethnicity, education, child age, or child sex (all P > 0.05). Our national surveys of parents and physicians identified messages that could motivate HPV vaccination, even among parents disinclined to vaccinate their children. The lack of difference across demographic subgroups in parental endorsement may suggest that these messages can be used across these subgroups. Our findings support physicians' use of these messages with parents to help motivate uptake of this important cancer-preventing vaccine. Cancer Epidemiol Biomarkers Prev; 25(10); 1383-91. ©2016 AACR.


SNP haplotypes and allele frequencies show evidence for disruptive and balancing selection in the human leukocyte receptor complex.
The human leukocyte receptor complex (LRC) of Chromosome 19q13.4 encodes polymorphic and highly homologous genes that are expressed by cells of the immune system and regulate their function. There is an enormous diversity at the LRC, most particularly the variable number of killer cell immunoglobulin-like receptor (KIR) genes. KIR have been associated with several disease processes due to their interaction with polymorphic human leukocyte antigen class I molecules. We have assessed haplotype compositions, linkage disequilibrium patterns and allele frequencies in two Caucasoid population samples (n=54, n=100), using a composite of single-nucleotide polymorphism (SNP) markers and high-resolution, allele-specific molecular genotyping. Particular KIR loci segregated with SNP and other markers, forming two blocks that were separated by a region with a greater history of recombination. The KIR haplotype composition and allele frequency distributions were consistent with KIR having been subject to balancing selection (Watterson's F: P=0.001). In contrast, there was a high inter-population heterogeneity measure for the LRC-encoded leukocyte immunoglobulin-like receptor A3 (LILRA3), indicating pathogen-driven disruptive selection (Wright's FST=0.32). An assessment of seven populations representative of African, Asian and Caucasoid ethnic groups (total n=593) provided little evidence for long-range LRC haplotypes. The different natural selection pressures acting on each locus may have contributed to a lack of linkage disequilibrium between them.


Factors Associated with Midterm Visual Field Variability in Patients with Stable Glaucoma.
To evaluate factors associated with midterm visual field (VF) variability in stable glaucoma patients in Brazil. This retrospective observational study included 59 eyes of 39 stable glaucoma patients. Baseline data assessed were age, gender, educational level, intraocular pressure (IOP), central corneal thickness, best-corrected visual acuity, spherical equivalent, number of hypotensive eye drops, type of glaucoma, number of VFs performed, follow-up in years, lens status, visual field index (VFI) values from the last 5 VF (standard automated perimetry (SAP)) tests, the presence or absence of central scotoma in the VF test, and the level of glaucomatous damage according to the VF mean deviation (MD) index of the last VFs. The 5 latest VFI scores were used to calculate the mean, the standard deviation (SD), and the coefficient of variation (CV). We divided the eyes into 2 groups, being group 1 comprised by the 29 eyes presenting the lowest CV values and group 2 comprised by the 30 eyes presenting the highest CV values. GEE models were used to compare the CV and demographic and clinical parameters of all participants. Mean age of all subjects was 65.8 ± 10.1 years. 54.0% were women. Average SAP MD values for groups 1 and 2 were -2.8 ± 3.1 dB and -6.2 ± 4.1 dB, respectively (P=0.006). Average SAP VFI values for groups 1 and 2 were 95.6 ± 5.9% and 85.9 ± 11.3%, respectively (P=0.002). There was a statistically significant association between CV and SAP MD values (P=0.006). A worse SAP MD and VFI were associated with a higher CV. In addition, even adjusting for potential confounding factors (age and level of education), the association between CV and the SAP MD and between CV and VFI remained significant (P ≤ 0.010). Glaucomatous patients with worse VF sensitivity scores (both MD and VFI indices) present higher VF test variability.


The variation of PbTiO3 bandgap at ferroelectric phase transition.
Optical properties of the PbTiO3 thin films fabricated by chemical solution deposition have been measured with variable angle spectroscopic ellipsometry in the spectral range of 1-6 eV and in the temperature interval from room temperature to 950 K. The optical response functions and band gap energy were determined in the whole temperature range. The direct band gap varies from the value 3.88 eV at room temperature to the value 3.67 eV just above the phase transition. The temperature dependence of the film lattice parameters was also measured by x-ray and it shows a strong correlation with the band gap. The comparison of experimental data with ab initio electronic structure calculations simulating the temperature development of dielectric function and band gap is also presented.


In vitro glucuronidation of 3'-azido-3'-deoxythymidine by human liver. Role of UDP-glucuronosyltransferase 2 form.
The glucuronidation of 3'-azido-3'-deoxythymidine (AZT) by human liver microsomes and human hepatocytes in culture has been studied in vitro to determine the UDP-glucuronosyltransferase (UDPGT) form conceivably involved in the AZT biotransformation process. The glucuronide of AZT was preliminarily identified through hydrolysis by beta-D-glucuronidase. Brij 58 was shown to be the best activator of AZT glucuronidation by human liver microsomes, as it increased the rate of glucuronide formation 3-fold. The UDPGT activities toward AZT measured in 29 different microsomal fractions was slightly variable among samples (79 to 268 nmol/hr/mg protein). The apparent KM value for AZT glucuronidation was about 5 mM. We sought to determine if various known UDPGT activities (i.e. p-nitrophenol UDPGT, 4-hydroxybiphenyl UDPGT, and DT1-UDPGT) in 18 microsomal samples were correlated with AZT-UDPGT activity. Experiments revealed that only 4-hydroxybiphenyl UDPGT activity was strongly correlated (r = 0.815, p less than 0.001) with AZT-UDPGT activity, whereas no correlation was found for the other UDPGT activities. To determine the isozyme conceivably involved in AZT glucuronidation, we studied the effect of various compounds on AZT glucuronidation. AZT glucuronidation was inhibited by numerous substrates of the UDPGT2, form: morphine (Ki = 1.8 mM), 4-hydroxybiphenyl (Ki = 0.92 mM), and ketoprofen (Ki = 0.75 mM), but also oxazepam, codeine, and chloramphenicol. p-Nitrophenol appeared to be an inhibitor, whereas acetaminophen had no effect. Bilirubin, aspirin, cimetidine, and acyclovir did not inhibit AZT glucuronidation. Since all the inhibitors tested except p-nitrophenol are known to be glucuronidated by the UDPGT2 form, our results strongly suggest the involvement of this isozyme in AZT glucuronidation.(ABSTRACT TRUNCATED AT 250 WORDS)


Cell therapy using encapsulated cells producing endostatin.
Despite aggressive surgery and post-operative radiation and chemotherapy, the prognosis is poor for glioblastoma patients. Anti-angiogenic therapy with compounds such as endostatin could delay the onset of relapse. However, the short systemic half-life of this proteins as well as the blood-brain barrier makes the use of this therapy difficult for brain cancer patients. The aim of this project is to develop and implant genetically engineered producer cells secreting endostatin that are encapsulated in calcium cross-linked alginate gel beads. Encapsulation of cells within alginate gels has a potential as a sustained release system in addition to the fact that the encapsulation technology protects the cells from rejection by the immune system. Human embryonal kidney 293 cells have been transfected with the gene for endostatin. These cells have been encapsulated in calcium cross-linked alginate gels and optimized for the secretion of endostatin. Alginate gel beads implanted into rat brain have shown only a moderate loss in cell viability but extended endostatin release for periods of up to 12 months. Visualization of the anti-angiogenic effect on C6 rat glioma growth, tumor vasculature and microhemodynamics has been demonstrated by using intravital video microscopy. The data indicates that endostatin greatly affects tumor-associated microcirculation but does not appear to affect normal microcirculation. The local delivery of endostatin seems to specifically affect tumor-associated microvessels by reduction of the vessel density, diameter and functionality. Tumor cell migration and invasion was greatly reduced in the endostatin treated animals.


A possible decline in the incidence and severity of Behçet's disease: implications for an infectious etiology and oral health.
Behçet's disease (BD) is a systemic and inflammatory disorder mainly present along the ancient Silk Road, from the Mediterranean to East Asia. A wide range of prevalence figures (0.1-420/100.000) are reported for BD, also among populations of similar ethnicity living in different countries. Recently, a decline of the incidence of BD and a change of the disease spectrum to less severe mucocutaneous manifestations is reported from Japan, a genetically homogenous, affluent population with limited immigration. Among environmental factors, a change in atopy/allergic disorders and a decline in infections are two possible mechanisms for this epidemiological change. A shift in Th1/Th2 immune balance towards Th2-associated immune responses are possible, however 'hygiene hypothesis' associated with this approach does not explain the recent trend of the increase in Th1-associated disorders such as inflammatory bowel disease and multiple sclerosis in Western Countries. We hypothesise that a decline in oral infections, associated with the improvement in oral health in Japan, could be behind this decline. Better epidemiological studies in other populations will show whether this decline is a worldwide trend and may provide a better understanding of the environmental factors associated with the onset or relapses of BD, leading a way to new therapeutic approaches.


Rate constant for the reaction of OH with H2 between 200 and 480 K.
The rate constant for the reaction of OH radicals with molecular hydrogen was measured using the flash photolysis resonance-fluorescence technique over the temperature range of 200-479 K. The Arrhenius plot was found to exhibit a noticeable curvature. Careful examination of all possible systematic uncertainties indicates that this curvature is not due to experimental artifacts. The rate constant can be represented by the following expressions over the indicated temperature intervals: k(H2)(250-479 K) = 4.27 x 10(-13) x (T/298)2.406 x exp[-1240/T] cm3 molecule(-1) (s-1) above T = 250 K and k(H2)(200-250 K) = 9.01 x 10(-13) x exp[-(1526 +/- 70)/T] cm3 molecule(-1) s(-1) below T = 250 K. No single Arrhenius expression can adequately represent the rate constant over the entire temperature range within the experimental uncertainties of the measurements. The overall uncertainty factor was estimated to be f(H2)(T) = 1.04 x exp[50 x /(1/T) - (1/298)/]. These measurements indicate an underestimation of the rate constant at lower atmospheric temperatures by the present recommendations. The global atmospheric lifetime of H2 due to its reaction with OH was estimated to be 10 years.


G-fibre cell wall development in willow stems during tension wood induction.
Willows (Salix spp.) are important as a potential feedstock for bioenergy and biofuels. Previous work suggested that reaction wood (RW) formation could be a desirable trait for biofuel production in willows as it is associated with increased glucose yields, but willow RW has not been characterized for cell wall components. Fasciclin-like arabinogalactan (FLA) proteins are highly up-regulated in RW of poplars and are considered to be involved in cell adhesion and cellulose biosynthesis. COBRA genes are involved in anisotropic cell expansion by modulating the orientation of cellulose microfibril deposition. This study determined the temporal and spatial deposition of non-cellulosic polysaccharides in cell walls of the tension wood (TW) component of willow RW and compared it with opposite wood (OW) and normal wood (NW) using specific antibodies and confocal laser scanning microscopy and transmission electron microscopy. In addition, the expression patterns of an FLA gene (SxFLA12) and a COBRA-like gene (SxCOBL4) were compared using RNA in situ hybridization. Deposition of the non-cellulosic polysaccharides (1-4)-β-D-galactan, mannan and de-esterified homogalacturonan was found to be highly associated with TW, often with the G-layer itself. Of particular interest was that the G-layer itself can be highly enriched in (1-4)-β-D-galactan, especially in G-fibres where the G-layer is still thickening, which contrasts with previous studies in poplar. Only xylan showed a similar distribution in TW, OW, and NW, being restricted to the secondary cell wall layers. SxFLA12 and SxCOBL4 transcripts were specifically expressed in developing TW, confirming their importance. A model of polysaccharides distribution in developing willow G-fibre cells is presented.


Design and analysis of association studies using pooled DNA from large twin samples.
Evidence is mounting that multiple genes are involved in complex traits and that these each account for very small proportions of the overall phenotypic variance. Association studies of many markers in 1000s of individuals will be required to identify such genes. A number of large twin cohorts have already been collected and provide a valuable resource for carrying out studies that are robust to the effect of population stratification. Technologies based on microarrays will soon allow 1.000,000 SNPs to be typed at one time, however financial considerations prevent most researchers from using these approaches to genotype all individuals. Recently, microarrays have been shown to give accurate allele frequency measurements in pooled DNA samples and provide a simple way to select the best markers for individual genotyping. This drastically reduces the cost and workload of large scale association studies. One limitation of this methodology relates to the analytical procedures which have only been developed to allow comparison of two pools e.g. case/control pools. In this paper we use meta-regression to analyze pooled DNA data allowing the allele frequency in each pool to be related to the average quantitative phenotypic measure of the individuals whose DNA were used to construct the pools. Alongside this we describe a technique that can be used to determine the power for such studies. We present results from some preliminary investigations of different pooling strategies that can be applied to large twin samples and demonstrate that the method retains a large proportion of the power available from individual genotyping.


Vibrational properties of organic donor-acceptor molecular crystals: Anthracene-pyromellitic-dianhydride (PMDA) as a case study.
We establish a reliable quantum-mechanical approach to evaluate the vibrational properties of donor-acceptor molecular crystals. The anthracene-PMDA (PMDA = pyromellitic dianhydride) crystal, where anthracene acts as the electron donor and PMDA as the electron acceptor, is taken as a representative system for which experimental non-resonance Raman spectra are also reported. We first investigate the impact that the amount of nonlocal Hartree-Fock exchange (HFE) included in a hybrid density functional has on the geometry, normal vibrational modes, electronic coupling, and electron-vibrational (phonon) couplings. The comparison between experimental and theoretical Raman spectra indicates that the results based on the αPBE functional with 25%-35% HFE are in better agreement with the experimental results compared to those obtained with the pure PBE functional. Then, taking αPBE with 25% HFE, we assign the vibrational modes and examine their contributions to the relaxation energy related to the nonlocal electron-vibration interactions. The results show that the largest contribution (about 90%) is due to electron interactions with low-frequency vibrational modes. The relaxation energy in anthracene-PMDA is found to be about five times smaller than the electronic coupling.


Comparison of the knowledge, attitude, and practices of doctors, nurses, and pharmacists regarding the use of expired and disposal of unused medicines in Delhi.
The use of out-of-date medicines and improper disposal of pharmaceutical products harm consumers and environment. To limit pharmaceutical load on the environment, it is important that healthcare professionals (HCPs) are aware of and have good practices toward their disposal. A pretested instructor administered questionnaire containing 21 questions was used to assess knowledge, attitude, and practices (KAP) of 300 HCPs toward use and disposal of unwanted/expired medicines. Nurses (76%) had significantly (P < 0.01) better knowledge over doctors regarding the methods of drug disposal, whereas both doctors (63%) and nurses (64%) knew significantly (P < 0.05) more than pharmacists about type of formulations not to be incinerated. The majority (88%) of overall respondents were unaware of validity of dispensing medicines before 15 days of mentioned expiry date. A large proportion of HCPs had a positive attitude toward checking the expiry date of medicines (96%), and a significant proportion of doctors and nurses believed that drug disposal is everyone's responsibility. Only 8% of the HCPs received unused/expired drugs back from the consumers. Nurses (59%) had better (P < 0.01) practice than pharmacists regarding the disposal of expired medicines. Nearly half (44%) of all HCPs agreed that there existed a system for removing expiry and unused medicines from the inventory. The KAP of HCPs toward use and disposal of out-of-date and unused medicines need upgrading. The existing guidelines should be implemented to ensure patients' safety and to preserve the ecosystem.


One Egg per Day Improves Inflammation when Compared to an Oatmeal-Based Breakfast without Increasing Other Cardiometabolic Risk Factors in Diabetic Patients.
There is concern that egg intake may increase blood glucose in patients with type 2 diabetes mellitus (T2DM). However, we have previously shown that eggs reduce inflammation in patients at risk for T2DM, including obese subjects and those with metabolic syndrome. Thus, we hypothesized that egg intake would not alter plasma glucose in T2DM patients when compared to oatmeal intake. Our primary endpoints for this clinical intervention were plasma glucose and the inflammatory markers tumor necrosis factor (TNF)-α and interleukin 6 (IL-6). As secondary endpoints, we evaluated additional parameters of glucose metabolism, dyslipidemias, oxidative stress and inflammation. Twenty-nine subjects, 35-65 years with glycosylated hemoglobin (HbA1c) values <9% were recruited and randomly allocated to consume isocaloric breakfasts containing either one egg/day or 40 g of oatmeal with 472 mL of lactose-free milk/day for five weeks. Following a three-week washout period, subjects were assigned to the alternate breakfast. At the end of each period, we measured all primary and secondary endpoints. Subjects completed four-day dietary recalls and one exercise questionnaire for each breakfast period. There were no significant differences in plasma glucose, our primary endpoint, plasma lipids, lipoprotein size or subfraction concentrations, insulin, HbA1c, apolipoprotein B, oxidized LDL or C-reactive protein. However, after adjusting for gender, age and body mass index, aspartate amino-transferase (AST) (p < 0.05) and tumor necrosis factor (TNF)-α (p < 0.01), one of our primary endpoints were significantly reduced during the egg period. These results suggest that compared to an oatmeal-based breakfast, eggs do not have any detrimental effects on lipoprotein or glucose metabolism in T2DM. In contrast, eggs reduce AST and TNF-α in this population characterized by chronic low-grade inflammation.


Brazilian adolescents infected by HIV: epidemiologic characteristics and adherence to treatment.
Over the last 3 decades since the first AIDS cases appeared, we have witnessed great progress in therapeutic methodologies that have transformed the evolution of the disease from debilitating and fatal, into chronic and controllable. HIV-infected children are arriving at adolescence and bringing specific challenges, not only to themselves, but also to their families and caregivers. This retrospective study sets forth epidemiological and treatment characteristics of 46 HIV-infected adolescents followed in a specialized university service relating said characteristics to therapy adherence assessed through a combination of three indirect methods. Therapy adherence did not reveal any association with either epidemiologic characteristics regarding age, sex, school level, household composition, age at diagnosis, mode of infection, knowledge of diagnosis, treatment time, or initial antiretroviral scheme. Patients with good therapy adherence presented lower viral load and used a smaller number of antiretroviral schemes.


Species-wide distribution of highly polymorphic minisatellite markers suggests past and present genetic exchanges among house mouse subspecies.
Four hypervariable minisatellite loci were scored on a panel of 116 individuals of various geographical origins representing a large part of the diversity present in house mouse subspecies. Internal structures of alleles were determined by minisatellite variant repeat mapping PCR to produce maps of intermingled patterns of variant repeats along the repeat array. To reconstruct the genealogy of these arrays of variable length, the specifically designed software MS_Align was used to estimate molecular divergences, graphically represented as neighbor-joining trees. Given the high haplotypic diversity detected (mean He = 0.962), these minisatellite trees proved to be highly informative for tracing past and present genetic exchanges. Examples of identical or nearly identical alleles were found across subspecies and in geographically very distant locations, together with poor lineage sorting among subspecies except for the X-chromosome locus MMS30 in Mus mus musculus. Given the high mutation rate of mouse minisatellite loci, this picture cannot be interpreted only with simple splitting events followed by retention of polymorphism, but implies recurrent gene flow between already differentiated entities. This strongly suggests that, at least for the chromosomal regions under scrutiny, wild house mouse subspecies constitute a set of interrelated gene pools still connected through long range gene flow or genetic exchanges occurring in the various contact zones existing nowadays or that have existed in the past. Identifying genomic regions that do not follow this pattern will be a challenging task for pinpointing genes important for speciation.


Evolutionary strata in a small mating-type-specific region of the smut fungus Microbotryum violaceum.
DNA sequence analysis and genetic mapping of loci from mating-type-specific chromosomes of the smut fungus Microbotryum violaceum demonstrated that the nonrecombining mating-type-specific region in this species comprises approximately 25% ( approximately 1 Mb) of the chromosome length. Divergence between homologous mating-type-linked genes in this region varies between 0 and 8.6%, resembling the evolutionary strata of vertebrate and plant sex chromosomes.


Adolescent caffeine consumption and self-reported violence and conduct disorder.
Caffeine is the most widely used psychoactive substance in the world and currently the only one legally available to children and adolescents. The sale and use of caffeinated beverages has increased markedly among adolescents during the last decade. However, research on caffeine use and behaviors among adolescents is scarce. We investigate the relationship between adolescent caffeine use and self-reported violent behaviors and conduct disorders in a population-based cross-sectional sample of 3,747 10th grade students (15-16 years of age, 50.2 % girls) who were enrolled in the Icelandic national education system during February 2012. Through a series of multiple regression models, while controlling for background factors, Attention Deficit Hyperactivity Disorder symptoms and current medication and peer delinquency, and including measures on substance use, our findings show robust additive explanatory power of caffeine for both violent behaviors and conduct disorders. In addition, the association of caffeine to the outcomes is significantly stronger for girls than boys for both violent behaviors and conduct disorders. Future studies are needed to examine to what extent, if at all, these relationships are causal. Indication of causal connections between caffeine consumption and negative outcomes such as those reported here would call into question the acceptability of current policies concerning the availability of caffeine to adolescents and the targeting of adolescence in the marketing of caffeine products.


Real life experience confirms sustained response to long-term biologics and switching in ankylosing spondylitis.
To investigate the long-term response to biological therapies in AS in a real life clinical setting and to quantify non-response and response to 'switching' therapies in these cases. All patients prescribed TNF-blocking therapies for AS between 1999 and 2006 were studied. Response was evaluated using Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing Spondylitis Functional Index (BASFI) and CRP results. A total of 113 patients (84 males: 29 females, mean age 45 yrs, median disease duration 16 yrs, 87% HLA-B27 positive) were identified. At baseline they had a mean BASDAI of 6.57, BASFI 6.57 and CRP of 31 g/dl. At the end of follow-up, these values had reduced to mean BASDAI of 3.12, BASFI 4.16 and CRP of 7 g/dl. Improvements were sustained for 24 months and beyond with no loss of effect. Only nine patients (8%) suffered side-effects leading to cessation or switching of first-line therapy and non-response occurred in 15 patients (13%) in the long term. Fifteen patients (13%) switched to a second drug and 14 of these (93%) had a significant and sustained response. Outcomes were similar regardless of drug used, duration of disease and HLA-B27 status. Treatment of active AS with TNF blockers according to the British Society of Rheumatology guidelines leads to a sustained response for over 2 yrs with most patients tolerating the drugs well. The rate of non-response is significantly lower than that seen in RA and nearly all of these patients respond well to a second-line agent.


Neural Categorization of Vibrotactile Frequency in Flutter and Vibration Stimulations: An fMRI Study.
As the use of wearable haptic devices with vibrating alert features is commonplace, an understanding of the perceptual categorization of vibrotactile frequencies has become important. This understanding can be substantially enhanced by unveiling how neural activity represents vibrotactile frequency information. Using functional magnetic resonance imaging (fMRI), this study investigated categorical clustering patterns of the frequency-dependent neural activity evoked by vibrotactile stimuli with gradually changing frequencies from 20 to 200 Hz. First, a searchlight multi-voxel pattern analysis (MVPA) was used to find brain regions exhibiting neural activities associated with frequency information. We found that the contralateral postcentral gyrus (S1) and the supramarginal gyrus (SMG) carried frequency-dependent information. Next, we applied multidimensional scaling (MDS) to find low-dimensional neural representations of different frequencies obtained from the multi-voxel activity patterns within these regions. The clustering analysis on the MDS results showed that neural activity patterns of 20-100 Hz and 120-200 Hz were divided into two distinct groups. Interestingly, this neural grouping conformed to the perceptual frequency categories found in the previous behavioral studies. Our findings therefore suggest that neural activity patterns in the somatosensory cortical regions may provide a neural basis for the perceptual categorization of vibrotactile frequency.


Identification of bovine CD14.
Six monoclonal antibodies (mAbs) that clustered together in a statistical analysis of data submitted to the Third Workshop on Ruminant Leucocyte Antigens and mAb CC-G33 were tested for ability to stain COS-7 cells transfected with cDNA encoding human CD14. Only mAb CC-G33 recognised the human molecule. The six mAbs were compared with mAb CC-G33 by flow cytometry and three were shown to be directed against bovine CD14.


Up-regulation of sucrose synthase and UDP-glucose pyrophosphorylase impacts plant growth and metabolism.
The effects of the overexpression of sucrose synthase (SuSy) and UDP-glucose pyrophosphorylase (UGPase) on plant growth and metabolism were evaluated in tobacco (Nicotiana tabacum cv. Xanthi). T(1) transgenic plants expressing either gene under the control of a tandem repeat cauliflower mosaic virus 35S promoter (2x35S) or a xylem-localized 4CL promoter (4-coumarate:CoA ligase; 4CL) were generated, and reciprocally crossed to generate plants expressing both genes. Transcript levels, enzyme activity, growth parameters, fibre properties and carbohydrate content of stem tissue were quantified. The expression profiles of both genes confirmed the expression pattern of the promoters: 2x35S expressed more strongly in leaves, while 4CL expression was highest in stem tissue. In-depth plant characterization revealed that the single-transgene lines showed significant increases in the height growth compared with corresponding control lines. The double-transgene plants demonstrated an additive effect, proving to be even taller than the single-transgene parents. Several of these lines had associated increases in soluble sugar content. Although partitioning of storage carbohydrates into starch or cellulose was not observed, the increased height growth and increases in soluble carbohydrates suggest a role for SuSy as a marker in sink strength and lend credit to the function of UGPase in a similar role. The up-regulation of these two genes, although not increasing the percentage cellulose content, was effective in increasing the total biomass, and thus the overall cellulose yield, from a given plant.


Locomotor mechanics during early life history: effects of size and ontogeny on fast-start performance of salmonid fishes
Fast-start locomotor behavior is important for escaping from predators and for capturing prey. To examine the effects of size and other aspects of developmental morphology on fast-start performance, the kinematics of the fast-start escape behavior were studied through early post-hatching development in three salmonid species: chinook salmon (Oncorhynchus tshawytscha), coho salmon (Oncorhynchus kisutch) and brown trout (Salmo trutta). These three species, while morphologically and developmentally similar, hatch and mature at different sizes (total length). Comparison of these species shows that some fast-start performance variables, including stage duration, maximum velocity and maximum acceleration, are highly dependent on ontogenetic state, while another, the overall distance traveled during stage 2, scales with total body length. Brown trout were studied from hatching into the juvenile development period. Aspects of fast-start performance peak at the end of yolk-sac absorption (the end of the eleutheroembryo phase) when the fish reaches the juvenile period. At this time, the durations of the fast-start stages are at their minima, and maximum velocity and maximum acceleration are at their highest levels relative to body length. Thus, escape behavior reaches its maximum size-specific performance at a relatively small size, just as the fish absorbs its yolk sac and begins to search for food. This peak in fast-start performance occurs during a life history period in which fast-start ability is likely to be particularly important for survival.


Vertebral Malalignment among Male Dentists with Work-related Musculoskeletal Pain in the United Arab Emirates.
Musculoskeletal pain (MSP) is a highly prevalent and debilitating disorder among dentists, often leading to early retirement. Poor ergonomics in the dental practice not only causes MSP, but can also lead to postural deviations as a result of an imbalance in muscle function. The aim of this study was to observe the relationship between MSP and vertebral deviations among male dentists in the United Arab Emirates (UAE). A total of 60 male dentists participated in this cross-sectional study. Standardized Nordic questionnaire (SNQ) was used to record MSP and PostureScreen Mobile® (PSM) application was used to observe postural deviations in the vertebral regions, including the neck, shoulders, upper back, lower back, and hips. About 83% of participants were suffering from MSP, with the lower back as the most commonly reported region (29%). The pain characteristics reported were chronicity (63%) and dullness (72%). As for postural deviations, the most common site recorded was the neck region (72%). The correlation between the regions of deviation and the regions of pain showed that the shoulder region was more significant than other regions. Musculoskeletal pain and vertebral malalignment are common among practicing dentists, probably because of bad postural habits and lack of awareness. Preventive strategies to minimize the risk of developing these health problems are highly recommended. Work-related MSP (WRMSP) is a significant health problem among dentists and may be linked to the later development of vertebral deviations. Posture screen analysis is a quick, yet objective postural and movement screening tool in which findings can quickly be rendered to prospective dentists, so that custom postural and functional exercises may be correctly practiced.


Actigraphy for measurements of sleep in relation to oncological treatment of patients with cancer: a systematic review.
Sleep disturbances are a prevalent and disabling problem for patients with cancer. Sleep disturbances are present throughout the cancer trajectory, especially during oncological treatment. Previously sleep disturbances have primarily been quantified with subjective rating scales. Actigraphy is an easy to use, non-invasive method for objective measurement of sleep. We systematically reviewed the literature for studies using actigraphy to measure sleeping habits of patients with cancer, undergoing oncological treatment. Our study furthermore reviewed studies with interventions designed to reduce sleep disturbances in the patient population. 19 studies were included in the final review of which 13 had a descriptive study design and six included some kind of intervention. The studies were primarily performed in patients with breast cancer undergoing chemotherapy. We found that sleep disturbances are prevalent, and persistent in patients with cancer. The sleep disturbances seem to be aggravated by chemotherapy treatment and accumulate as the treatment continues. Sleep disturbances need further attention among clinicians working with patients with cancer.


Antigen presentation by murine dendritic cells is nuclear factor-kappa B dependent both in vitro and in vivo.
Antigen presentation is a key rate-limiting step in the immune response. Dendritic cells (DCs) have been reported to be the most potent antigen-presenting cells for naïve T cells, but little is known about the biochemical pathways that regulate this function. We here demonstrate that mature murine DC can be infected with adenovirus at high efficiency (>95%) and that an adenovirus transferring the endogenous inhibitor IkappaBalpha blocks nuclear factor-kappa B (NF-kappaB) function in murine DC. This result indicates that antigen presentation in the mixed leucocyte reaction is NF-kappaB dependent, confirming data with human DC in vitro. However, the importance of this finding depends on verifying that this is true also in vivo. Using delayed type hypersensitivity with allogeneic cells, we show that NF-kappaB inhibition had a marked immunosuppressive effect in vivo. These results thus establish NF-kappaB as an effective target for blocking DC antigen presentation and hence inhibiting T-cell-dependent immune responses. This finding has potential implications for the development of therapeutic agents for use in various pathological conditions of the immune system, including allergy and autoimmunity, and also in transplantation.


Implication of Nrf2/HO-1 pathway in the coloprotective effect of coenzyme Q10 against experimentally induced ulcerative colitis.
Given the increased incidence of ulcerative colitis worldwide, the current study was designed to investigate the coloprotective potential of CoQ10 against experimentally induced ulcerative colitis (UC) and specify the implicated mechanisms. Ulcerative colitis was induced by intracolonic instillation of [2 ml, 3% v/v acetic acid (AA)]. Rats in the different experimental groups received CoQ10 (10 or 30 and 100 mg/kg, orally) for eight consecutive days, either in a protective or curative regimen. Intracolonic AA instillation significantly increased colon/body weight index, colon weight/colon length ratio, clinical evaluation and macroscopic scoring of UC, serum LDH, C-reactive protein and decreased the serum total antioxidant capacity. Colon MDA, TNF-α and calcium content significantly increased as well, with concomitant reduction in colon GSH, SOD, CAT, Nrf2 and HO-1 contents. Moreover, immunohistochemical staining of colon specimen revealed increased expression of caspase-3 with significant histopathological changes. Coenzyme Q10 suppressed the release of inflammatory biomarkers and restored oxidants/antioxidants hemostasis. In a dose-dependent manner, CoQ10 significantly decreased colon/body weight index, colon weight/colon length ratio, clinical evaluation and macroscopic scoring of UC, serum LDH, C-reactive protein, colon MDA, TNF-α, caspase-3 expression and increased the serum total antioxidant capacity. Colon GSH, SOD, CAT, Nrf2 and HO-1 contents significantly increased. Moreover, coenzyme Q10 significantly preserved tissue histopathological architecture. It appears that the coloprotective effect of CoQ10 was calcium-independent. Coenzyme Q10 dose-dependently protects against AA-induced UC mainly via modulation of Nrf2/HO-1 and caspase-3 pathways. Antioxidant, anti-inflammatory and anti-apoptotic properties of CoQ10 are implicated in its observed therapeutic benefit.


Strongyloides papillosus causes sudden death in weaned calves on New York dairies.
Outbreaks of sudden death in apparently healthy weaned dairy calves due to Strongyloides papillosus parasitism were diagnosed on 2 separate and independent New York (NY) dairies. Most calves were found dead; however, 1 calf observed while dying showed signs of tachycardia, tachypnea, vocalization, and convulsions shortly before death. In 6 affected heifers that underwent post-mortem examination, precocious bilaterally symmetric mammary gland enlargement was seen. A portion of their parasitized living cohorts also demonstrated similar mammary gland enlargement. A diagnosis of S papillosus hyperinfection was made based upon the presence of high numbers of S papillosus ova in feces, and confirmation by S papillosus-specific PCR assays. Consistent histopathological findings in affected calves included generalized mammary gland vascular congestion, interstitial edema and hemorrhage with ductal hyperplasia. Mild multifocal cardiomyocyte degeneration was found in 5 of 14 calves examined. Factors believed to contribute to the parasite's environmental amplification and host hyperinfection included group housing on wood shavings and high environmental temperatures and humidity. Treatment of calves with doramectin pour-on stopped mortality and resolved the udder enlargement. Similar outbreaks have previously been described in Japan and South Bohemia (Czech Republic), where researchers hypothesized that sudden death may be due to fatal arrhythmia caused by a parasite-associated cardiotoxin. This report highlights the importance of including S papillosus among the differential diagnoses for sudden death alone or together with precocious udder enlargement in calves kept in confinement housing.


The risk of congestive heart failure in rheumatoid arthritis: a population-based study over 46 years.
It is hypothesized that the systemic inflammation associated with rheumatoid arthritis (RA) promotes an increased risk of cardiovascular (CV) morbidity and mortality. We examined the risk and determinants of congestive heart failure (CHF) in patients with RA. We assembled a population-based, retrospective incidence cohort from among all individuals living in Rochester, Minnesota, in whom RA (defined according to the American College of Rheumatology 1987 criteria) was first diagnosed between 1955 and 1995, and an age- and sex-matched non-RA cohort. After excluding patients in whom CHF occurred before the RA index date, all subjects were followed up until either death, incident CHF (defined according to the Framingham Heart Study criteria), migration from the county, or until January 1, 2001. Detailed information from the complete medical records (including all inpatient and outpatient care provided by all local providers) regarding RA, ischemic heart disease, and traditional CV risk factors was collected. Cox models were used to estimate the effect of RA on the development of CHF, adjusting for CV risk factors and/or ischemic heart disease. The study population included 575 patients with RA and 583 subjects without RA. The CHF incidence rates were 1.99 and 1.16 cases per 100 person-years in patients with RA and in non-RA subjects, respectively (rate ratio 1.7, 95% confidence interval [95% CI] 1.3-2.1). After 30 years of followup, the cumulative incidence of CHF was 34.0% in patients with RA and 25.2% in non-RA subjects (P< 0.001). RA conferred a significant excess risk of CHF (hazard ratio [HR] 1.87, 95% CI 1.47-2.39) after adjusting for demographics, ischemic heart disease, and CV risk factors. The risk was higher among patients with RA who were rheumatoid factor (RF) positive (HR 2.59, 95% CI 1.95-3.43) than among those who were RF negative (HR 1.28, 95% CI 0.93-1.78). Compared with persons without RA, patients with RA have twice the risk of developing CHF. This excess risk is not explained by traditional CV risk factors and/or clinical ischemic heart disease.


Cancer fatalism and poor self-rated health mediate the association between socioeconomic status and uptake of colorectal cancer screening in England.
Little is known about the psychological predictors of colorectal screening uptake in England and mediators of associations between uptake and socioeconomic status (SES). This study tested the hypotheses that although higher threat and efficacy beliefs, lower cancer fatalism, lower depression, and better self-rated health would predict higher screening uptake, only efficacy beliefs, fatalism, depression, and self-rated health would mediate associations between uptake and SES. Data from 529 adults aged 60 to 69 who had completed a postal survey in 2005-2006 were linked with data on fecal occult blood test (FOBt) uptake recorded at the screening "hub" following its introduction in 2007, resulting in a prospective study. Screening uptake was 56% and was higher among people with higher SES, better self-rated health, higher self-efficacy beliefs, and lower cancer fatalism in univariate analyses. Path analysis on participants with complete data (n = 515) showed that both better self-rated health and lower cancer fatalism were directly associated with higher uptake of FOBt screening and significantly mediated pathways from SES to uptake. Lower depression only had an indirect effect on uptake through better self-rated health. Efficacy beliefs did not mediate the relationship between SES and uptake. SES differences in uptake of FOBt in England are partially explained by differences in cancer fatalism, self-rated health, and depression. This is one of only a few studies to examine mediators of the relationship between SES and screening uptake, and future research could test the effectiveness of interventions to reduce fatalistic beliefs to increase equality of uptake.


A time-series prediction approach for feature extraction in a brain-computer interface.
This paper presents a feature extraction procedure (FEP) for a brain-computer interface (BCI) application where features are extracted from the electroencephalogram (EEG) recorded from subjects performing right and left motor imagery. Two neural networks (NNs) are trained to perform one-step-ahead predictions for the EEG time-series data, where one NN is trained on right motor imagery and the other on left motor imagery. Features are derived from the power (mean squared) of the prediction error or the power of the predicted signals. All features are calculated from a window through which all predicted signals pass. Separability of features is achieved due to the morphological differences of the EEG signals and each NNs specialization to the type of data on which it is trained. Linear discriminant analysis (LDA) is used for classification. This FEP is tested on three subjects off-line and classification accuracy (CA) rates range between 88% and 98%. The approach compares favorably to a well-known adaptive autoregressive (AAR) FEP and also a linear AAR model based prediction approach.


Social Mishap Exposures for Social Anxiety Disorder: An Important Treatment Ingredient.
Conventional cognitive-behavioral therapy for social anxiety disorder, which is closely based on the treatment for depression, has been shown to be effective in numerous randomized placebo-controlled trials. Although this intervention is more effective than waitlist control group and placebo conditions, a considerable number of clients do not respond to this approach. Newer approaches include techniques specifically tailored to this particular population. One of these techniques, social mishap exposure practice, is associated with significant improvement in treatment gains. We will describe here the theoretical framework for social mishap exposures that addresses the client's exaggerated estimation of social cost. We will then present clinical observations and outcome data of a client who underwent treatment that included such social mishap exposures. Findings are discussed in the context of treatment implications and directions for future research.


Content and bibliometric analyses of the Journal of Manual & Manipulative Therapy.
Article characteristics and trends have been elucidated for other physical therapy-focused journals using content and bibliometric analysis. These findings are important for assessing the current state of a journal and for guiding future publication of research. To date, these analyses have not been performed for the Journal of Manual & Manipulative Therapy (JMMT). To describe content and trends for articles published in JMMT over a 20-year period (1993-2012). Journal articles were coded using previously-established domains (article type, participant type, research design, study purpose, and clinical condition). Total publications and proportion of publications based on domain were described. Articles specific to manual therapy intervention were examined and compared to data from other physical therapy-focused journals. Impact by citation and author was examined using bibliometric software. Journal of Manual & Manipulative Therapy was found to have a recent acceleration in the number of articles published annually. Over time, topical reviews have decreased in favor of research reports. However, rigorous study designs have represented only a small portion of total journal content, and case reports have maintained a consistent publication presence. Manual therapy intervention articles in JMMT are predominantly case designs, however are similar in characteristics to manual therapy intervention articles published in other physical therapy-focused journals. For JMMT articles overall and manual therapy intervention articles across journals, young to middle-aged symptomatic adults with low back and/or neck pain were the most common study participants. Increases in the number of papers and a move toward research reports were observed in JMMT over the 20-year period. Considerations for the future were outlined, including the publication of articles with more rigorous research designs. Manual therapy research for adolescents and older adults and for upper and lower extremity conditions should also be considered as priorities for the future.


Intrapatient escape in the A*0201-restricted epitope SLYNTVATL drives evolution of human immunodeficiency virus type 1 at the population level.
The hypothesis that the intrapatient emergence of cytotoxic T-lymphocyte escape variants contributes to the evolution of human immunodeficiency virus type 1 at the population (interpatient) level was tested using the HLA-A*0201-restricted gag p17 epitope SLYNTVATL. Using a simple experimental design, we investigated the evolutionary processes operating within this epitope among patients while compensating for the confounding influence of intrapatient natural selection. Using this approach, we revealed a pattern of A*0201-driven escape within patients, followed by the sustained transmission of these escape variants among patients irrespective of their HLA type.


A host-selective toxin of Pyrenophora tritici-repentis, Ptr ToxA, induces photosystem changes and reactive oxygen species accumulation in sensitive wheat.
Ptr ToxA (ToxA) is a proteinaceous necrotizing host-selective toxin produced by Pyrenophora tritici-repentis, a fungal pathogen of wheat (Triticum aestivum). In this study, we have found that treatment of ToxA-sensitive wheat leaves with ToxA leads to a light-dependent accumulation of reactive oxygen species (ROS) that correlates with the onset of necrosis. Furthermore, the accumulation of ROS and necrosis could be inhibited by the antioxidant N-acetyl cysteine, providing further evidence that ROS production is required for necrosis. Microscopic evaluation of ToxA-treated whole-leaf tissue indicated that ROS accumulation occurs in the chloroplasts. Analysis of total protein extracts from ToxA-treated leaves showed a light-dependent reduction of the chloroplast protein RuBisCo. In addition, Blue native-gel electrophoresis followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis revealed that ToxA induces changes in photosystem I (PSI) and photosystem II (PSII) in the absence of light, and therefore, the absence of ROS. When ToxA-treated leaves were exposed to light, all proteins in both PSI and PSII were extremely reduced. We propose that ToxA induces alterations in PSI and PSII affecting photosynthetic electron transport, which subsequently leads to ROS accumulation and cell death when plants are exposed to light.


Effects of carbamazepine and cetirizine under an ocean acidification scenario on the biochemical and transcriptome responses of the clam Ruditapes philippinarum.
Several works evaluated the toxicity of pharmaceutical drugs and climate related changes in invertebrates but few explored the combined effects of both stressors, namely considering their mode of action (MoA). Carbamazepine (CBZ) and cetirizine (CTZ) are pharmaceutical drugs detected in the environment and the toxicity derived from the combined effects of these drugs with ocean acidification (OA) is poorly explored. Thus, the present study investigated the biochemical parameters related to an oxidative stress response and the transcription of genes related to the MoA of CBZ (1.0 μg/L) and CTZ (0.6 μg/L) in the clam Ruditapes philippinarum chronically exposed (28 days) to control (7.8) and low (7.5) pH conditions. The results obtained showed that despite the clams accumulated both drugs, at low pH the clams exposed to CTZ decreased drug concentration and BCF values (CTZ uptake: 2.0 ± 0.5 ng/g fresh weight; BCF: 3.8 ± 0.9) in comparison with clams exposed to control pH (CTZ uptake: 2.9 ± 0.3 ng/g fresh weight; BCF: 5.5 ± 0.6). No oxidative stress was induced by the exposure to CBZ or CTZ at each pH level, but the transcription of several genes related with the MoA (neurotransmission, immunity and biomineralization) was altered by low pH, drug exposure and the combination of both stressors. At both pH conditions, CBZ increased the transcription of GABA receptor gene (neurotransmission) and CTZ led to a decrease of Perlucin gene (biomineralization) transcription. The transcription of MyD88 gene (immunity) decreased at low pH (7.5) combined with drug exposure (CBZ or CTZ). Thus, it was highlighted that the interaction of drug exposure and low pH conditions can change bivalves' sensitivity to drugs or alter drugs toxicity.


[The role of some cellular components of bacterial parasites in determining the incidence of tuberculosis: studies on mycobacterial antigens, with special reference to mycobacterial immunoreactive ribosomal and secreted proteins].
Tuberculosis remains as major disease, affecting more than 20 million people. The elimination of the disease with vaccination, rapid diagnosis, and and efficient therapy is an important objective of our study. To realize the objective, the characterization of antigens is essential. We have chosen two kinds of antigens for our study, the ribosomal antigens and and an antigenic proteins secreted by mycobacteria. The biochemical and immunological characterization of ribosomal fraction was carried out. Ribosomal proteins were purified and assessed for DTH reaction. The N-terminal amino acids sequences were determined. Total structures of S19, S7 and S12 in 30S and L7/L12 in 50S subunits were elucidated. L7/L12 had 66% homology with analogue from S. griseus which showed GTPase activity in protein synthesis. This protein was secreted in culture medium and induced strong DTH. Secreted antigenic proteins are of great interest for us. Secreted antigens may be recognized rapidly by immune system and therefore may induce rapid and high level immune response. It is also expected that it may contain protective antigens, since live BCG protect disease more efficiently than heat killed BCG. We have determined and published the total structure of four proteins (MPB64, MPB70, MPB57 and alpha antigen). We attempted to utilize this antigen for the diagnosis and the design of vaccine. The structures of alpha antigens from M. avium, M. intracellulare, M. scrofulaceum, M. kansasii and BCG were determined and its potential for application to diagnosis was presented. Using the operon of M. kansasii, alpha antigen and V3 region of HIV-1 were expressed by recombinant BCG which induced CTL in mice.


A role for Saccharomyces cerevisiae Chk1p in the response to replication blocks.
Replication blocks and DNA damage incurred during S phase activate the S-phase and intra-S-phase checkpoint responses, respectively, regulated by the Atrp and Chk1p checkpoint kinases in metazoans. In Saccharomyces cerevisiae, these checkpoints are regulated by the Atrp homologue Mec1p and the kinase Rad53p. A conserved role of these checkpoints is to block mitotic progression until DNA replication and repair are completed. In S. cerevisiae, these checkpoints include a transcriptional response regulated by the kinase Dun1p; however, dun1Delta cells are proficient for the S-phase-checkpoint-induced anaphase block. Yeast Chk1p kinase regulates the metaphase-to-anaphase transition in the DNA-damage checkpoint pathway via securin (Pds1p) phosphorylation. However, like Dun1p, yeast Chk1p is not required for the S-phase-checkpoint-induced anaphase block. Here we report that Chk1p has a role in the intra-S-phase checkpoint activated when yeast cells replicate their DNA in the presence of low concentrations of hydroxyurea (HU). Chk1p was modified and Pds1p was transiently phosphorylated in this response. Cells lacking Dun1p were dependent on Chk1p for survival in HU, and chk1Delta dun1Delta cells were defective in the recovery from replication interference caused by transient HU exposure. These studies establish a relationship between the S-phase and DNA-damage checkpoint pathways in S. cerevisiae and suggest that at least in some genetic backgrounds, the Chk1p/securin pathway is required for the recovery from stalled or collapsed replication forks.


High Sporozoite Antibody Titers in Conjunction with Microscopically Detectable Blood Infection Display Signatures of Protection from Clinical Malaria.
Immunoepidemiological studies typically reveal slow, age-dependent acquisition of immune responses against Plasmodium falciparum sporozoites. Naturally acquired immunity against preerythrocytic stages is considered inadequate to confer protection against clinical malaria. To explore previously unrecognized antisporozoite responses, we measured serum levels of naturally acquired antibodies to whole Plasmodium falciparum sporozoites (Pfspz) and the immunodominant (NANP)5 repeats of the major sporozoite surface protein, circumsporozoite protein, in a well-characterized Kenyan cohort. Sera were sampled at the start of the malaria transmission season, and all subjects were prospectively monitored for uncomplicated clinical malaria in the ensuing 6 months. We used Kaplan-Meier analysis and multivariable regression to investigate the association of antisporozoite immunity with incidence of clinical malaria. Although naturally acquired humoral responses against Pfspz and (NANP)5 were strongly correlated (p < 0.0001), 37% of Pfspz responders did not recognize (NANP)5. The prevalence and magnitude of antisporozoite responses increased with age, although some high Pfspz responders were identified among children. Survival analysis revealed a reduced risk of and increased time to first or only episode of clinical malaria among Pfspz or (NANP)5 responders carrying microscopically detectable Plasmodium falciparum (Pf) parasitemia at the start of the transmission season (p < 0.03). Our Cox regression interaction models indicated a potentially protective interaction between high anti-Pfspz (p = 0.002) or anti-(NANP)5 (p = 0.001) antibody levels and microscopically detectable Pf parasitemia on the risk of subsequent clinical malaria. Our findings indicate that robust antisporozoite immune responses can be naturally acquired already at an early age. A potentially protective role of high levels of anti-Pfspz antibodies against clinical episodes of uncomplicated malaria was detected, suggesting that antibody-mediated preerythrocytic immunity might indeed contribute to protection in nature.


Comparative genomic and metabolic analysis of three Lactobacillus paracasei cheese isolates reveals considerable genomic differences in strains from the same niche.
Strains of Lactobacillus paracasei are present in many diverse environments, including dairy and plant materials and the intestinal tracts of humans and animals. Their adaptation to various niches is correlated to intra-species diversity at the genomic and metabolic level. In this study, we compared the genome sequences of three L. paracasei strains isolated from mature Cheddar cheeses, two of which (DPC4206 and DPC4536) shared the same genomic fingerprint by PFGE, but demonstrated varying metabolic capabilities. Genome sizes varied from 2.9 Mbp for DPC2071, to 3.09 Mbp for DPC4206 and 3.08 Mpb for DPC4536. The presence of plasmids was a distinguishing feature between the strains with strain DPC2071 possessing an unusually high number of plasmids (up to 11), while DPC4206 had one plasmid and DPC4536 harboured no plasmids. Each of the strains possessed specific genes not present in the other two analysed strains. The three strains differed in their abundance of sugar-converting genes, and in the types of sugars that could be used as energy sources. Genes involved in the metabolism of sugars not usually connected with the dairy niche, such as myo-inositol and pullulan were also detected, but strains did not utilise these sugars. The genetic content of the three strains differed in regard to specific genes for arginine and sulfur-containing amino acid metabolism and genes contributing to resistance to heavy metal ions. In addition, variability in the presence of phage remnants and phage protection systems was evident. The findings presented in this study confirm a considerable level of heterogeneity of Lactobacillus paracasei strains, even between strains isolated from the same niche.


[DSM-5: from 'somatoform disorders' to 'somatic symptom and related disorders'].
The DSM-IV somatoform disorder category was controversial and has undergone major changes in DSM-5. To provide a critical description of DSM-5 somatic symptoms and related disorders (SSD). To review the new classification system for somatic symptom and related disorders (SSD) as set out in DSM-5 RESULTS: Whereas the symptoms of somatoform disorder were always considered to be medically unexplained, in DSM-5 the symptoms of somatic symptom disorder can now sometimes be explained by a medical condition. The criterion is that the individual has a maladaptive reaction to a somatic symptom. In DSM-5 the terms somatisation disorder, pain disorder and undifferentiated somatoform disorder have been discarded, whereas factitious disorder as well as psychological factors affecting other medical conditions have been added to somatic symptom disorder and other disorders. Conversion disorder remains as it was in DSM-IV. Hypochondriasis has been renamed 'illness anxiety disorder'. The new description of somatic symptom disorder in DSM-5 represents a big step forwards, because the decision has been made to use, for classification, a positive criterion, namely maladaptive reaction to a somatic symptom, instead of the earlier negative criterion, namely that the symptoms should be medically unexplained. Before attaching the ssd label to an illness, the clinician will have to weigh up the clinical consequences of this psychiatric diagnosis.


Recurrent acute transverse myelopathy associated with anticardiolipin antibodies.
In three patients with recurrent episodes of acute transverse myelopathy, spinal MR imaging during each episode showed areas of hyperintensity on proton density- and T2-weighted images with inconsistent contrast enhancement. Cranial MR imaging, laboratory screenings, and CSF analysis showed only increased titers of anticardiolipin antibodies. Although a causative role in neurologic conditions has not been established conclusively, an association between these antibodies and acute transverse myelopathy and its recurrences cannot be ignored.


Audit of antenatal screening for syphilis and HIV in migrant and refugee women on the Thai-Myanmar border: a descriptive study.
The antenatal prevalence of syphilis and HIV/AIDS in migrants and refugees is poorly documented. The aim of this study was to audit the first year of routine syphilis screening in the same population and reassess the trends in HIV rates. From August 2012 to July 2013, 3600 pregnant women were screened for HIV (ELISA) and syphilis (VDRL with TPHA confirmation) at clinics along the Thai-Myanmar border. Seroprevalence for HIV 0.47% (95% CI 0.30-0.76) (17/3,599), and syphilis 0.39% (95% CI 0.23-0.65) (14/3,592), were low. Syphilis was significantly lower in refugees (0.07% 95% CI 0.01-0.38) (1/1,469), than in migrants (0.61% 95% CI 0.36-1.04) (13/2,123). The three active (VDRL≥1:8 and TPHA reactive) syphilis cases with VDRL titres of 1:32 were easy to counsel and treat. Women with low VDRL titres (>75% were < 1:8) and TPHA reactive results, in the absence of symptoms and both the woman and her husband having only one sexual partner in their lifetime, and the inability to determine the true cause of the positive results presented ethical difficulties for counsellors. As HIV and syphilis testing becomes available in more and more settings, the potential impact of false positive results should be considered, especially in populations with low prevalence for these diseases. This uncertainty must be considered in order to counsel patients and partners accurately and safely about the results of these tests, without exposing women to increased risk for abuse or abandonment. Our findings highlight the complexities of counselling patients about these tests and the global need for more conclusive syphilis testing strategies.


From patient to model: stereolithographic modeling of the cerebral vasculature based on rotational angiography.
We developed a method to produce tubular in vitro models of the cerebral vessels from real patient data. Three-dimensional data sets obtained from patients undergoing rotational angiography were used for stereolithographic biomodeling by using rapid prototyping technology. In a second step, tubular reproductions of the cerebral vessels were obtained by using the lost-wax technique. These reproductions can be useful for hemodynamic research and for the development and preclinical evaluation of new endovascular treatment methods.


Near-fatal asthma in South Australia: descriptive features and medication use.
Self-reported prior morbidity levels and medication use among survivors of a near-fatal asthma attack (NFA) were studied. To identify deficiencies in asthma management and opportunities for intervention. A hundred and twenty-seven consecutive patients aged 15 years or more presenting with a NFA to accident and emergency departments of teaching hospitals were interviewed. High levels of morbidity due to asthma were reported. Most cases (79%) reported symptoms occurring at least weekly in the three months before their NFA. A mean of 20.8 days was reportedly lost from work, school or other usual daily activity in the 12 months before these events. Regular use of beta agonist as nebuliser solution was reported by 27% of cases, increasing to 34.5% in response to increased symptoms, while 41% reported use of nebulised beta agonist in response to the NFA event. Less than half of all cases (46%) reported using an inhaled corticosteroid on a regular basis. Oral corticosteroids were used by 33% of cases at times of increased symptoms in the preceding 12 months. However, only 7% of cases reported initiating or increasing oral corticosteroids at the time of the NFA. Despite high levels of prior asthma morbidity, regular preventive inhaled corticosteroid use was not widespread in this series of NFA asthmatics. By comparison, over-reliance on regular beta agonist medication was common. Oral corticosteroids were rarely commenced in response to the NFA.


Regulation of conductive chloride transport in human placental brush border membrane vesicles.
Conductive chloride transport in the brush border of the human placental epithelium has been determined by measurement of radioactive chloride flux (influx and efflux) and anion diffusion potentials. This conductance is inhibited specifically by low micromolar concentrations of arachidonic acid whereas the K+ permeability of this membrane is increased by arachidonic acid. The conductive pathway for anions is inhibited by 9-AC and has the permeability sequence SCN > I > NO3 > Br > Cl > F > gluconate; it is also permeant to HCO3-. Functionally the control of these anion and cationic conductances by arachidonic acid will be important in regulating epithelial transport and nutrient delivery to the fetus since they will set the value of the resting potential across the apical surface of the syncytiotrophoblast.


The systemic autoinflammatory disorders for dermatologists. Part 2: disease examples.
The systemic autoinflammatory disorders (SAIDS) or periodic fever syndromes are disorders of innate immunity, which can be inherited or acquired. They are almost all very rare and easily overlooked; typically, patients will have seen multiple specialities prior to diagnosis, so a high level of clinical suspicion is key. It is important to note that these are 'high-value' diagnoses as the majority of these syndromes can be very effectively controlled, dramatically improving quality of life and providing protection against the development of irreversible complications such as AA amyloidosis. In Part 1 of this review, we took an overview of SAIDS and described the common features; in this article, we take a more in-depth look at the better recognized or more dermatologically relevant conditions.


What will Become of the Taxpayer Investment in Public Cord Blood Stem Cell Banking?
Cord Blood (CB) is a unique and readily available source of hematopoietic stem cells for transplantation. CB also contains other types of stem cells, including endothelial stem cells and mesenchymal stem cells, that may prove useful in non-traditional clinical uses. Genetic and molecular analyses have demonstrated that CB stem cells lie somewhere between mature stem cells like those found in Bone Marrow (BM), and fetal stem cells. After 25 years of clinical experience, CB is now used in the same fashion as BM for all typical malignant and genetic diseases treated by bone marrow transplant. Due to the establishment of CB banks in the US and abroad, more than 35,000 CB transplants have been performed over the past 25 years. An average of 700-800 CB transplants are performed annually. In addition, CB is now used more frequently for regenerative medicine and tissue engineering applications. At first glance, it seems that everything could not be better with the public cord blood banks and the use of their samples in the clinic. However, a recent report by the Rand Corp. reviewed the US national cord blood stem cell banking program and detailed many ongoing problems. However, some details were omitted from the report that would shed some light on the causes of many of the problems. This paper will summarize the status of the public cord blood stem cell banking program in the US, detail the problems associated with the program that could jeopardize its existence and suggest possible solutions to resolve these issues.


Expression of a single-chain variable-fragment antibody against a Fusarium virguliforme toxin peptide enhances tolerance to sudden death syndrome in transgenic soybean plants.
Plants do not produce antibodies. However, plants can correctly assemble functional antibody molecules encoded by mammalian antibody genes. Many plant diseases are caused by pathogen toxins. One such disease is the soybean sudden death syndrome (SDS). SDS is a serious disease caused by the fungal pathogen Fusarium virguliforme. The pathogen, however, has never been isolated from diseased foliar tissues. Thus, one or more toxins produced by the pathogen have been considered to cause foliar SDS. One of these possible toxins, FvTox1, was recently identified. We investigated whether expression of anti-FvTox1 single-chain variable-fragment (scFv) antibody in transgenic soybean can confer resistance to foliar SDS. We have created two scFv antibody genes, Anti-FvTox1-1 and Anti-FvTox1-2, encoding anti-FvTox1 scFv antibodies from RNAs of a hybridoma cell line that expresses mouse monoclonal anti-FvTox1 7E8 antibody. Both anti-FvTox1 scFv antibodies interacted with an antigenic site of FvTox1 that binds to mouse monoclonal anti-FvTox1 7E8 antibody. Binding of FvTox1 by the anti-FvTox1 scFv antibodies, expressed in either Escherichia coli or transgenic soybean roots, was initially verified on nitrocellulose membranes. Expression of anti-FvTox1-1 in stable transgenic soybean plants resulted in enhanced foliar SDS resistance compared with that in nontransgenic control plants. Our results suggest that i) FvTox1 is an important pathogenicity factor for foliar SDS development and ii) expression of scFv antibodies against pathogen toxins could be a suitable biotechnology approach for protecting crop plants from toxin-induced diseases.


Recruitment of a ROC1-CUL1 ubiquitin ligase by Skp1 and HOS to catalyze the ubiquitination of I kappa B alpha.
Activation of the transcription factor NF-kappa B in response to proinflammatory stimuli requires the phosphorylation-triggered and ubiquitin-dependent degradation of the NF-kappa B inhibitor, I kappa B alpha. Here, we show the in vitro reconstitution of the phosphorylation-dependent ubiquitination of I kappa B alpha with purified components. ROC1, a novel SCF-associated protein, is recruited by cullin 1 to form a quatemary SCFHOS-ROC1 holenzyme (with Skp1 and the beta-TRCP homolog HOS). SCFHOS-ROC1 binds IKK beta-phosphorylated I kappa B alpha and catalyzes its ubiquitination in the presence of ubiquitin, E1, and Cdc34. ROC1 plays a unique role in the ubiquitination reaction by heterodimerizing with cullin 1 to catalyze ubiquitin polymerization.


Compensatory UTE/T2W Imaging of Inflammatory Vascular Wall in Hyperlipidemic Rabbits.
To obtain compensatory ultra-short echo time (UTE) imaging and T2-weighted (T2W) imaging of Watanabe heritable hyperlipidemic (WHHL) rabbits following dextran-coated magnetic nanocluster (DMNC) injection for the effective in vivo detection of inflammatory vascular wall. Magnetic nanoparticle was synthesized by thermal decomposition and encapsulated with dextran to prepare DMNC. The contrast enhancement efficiency of DMNC was investigated using UTE (repetition time [TR] = 5.58 and TE = 0.07 ms) and T2W (TR = 4000 and TE = 60 ms) imaging sequences. To confirm the internalization of DMNC into macrophages, DMNC-treated macrophages were visualized by cellular transmission electron microscope (TEM) and magnetic resonance (MR) imaging. WHHL rabbits expressing macrophage-rich plaques were subjected to UTE and T2W imaging before and after intravenous DMNC (120 μmol Fe/kg) treatment. Ex vivo MR imaging of plaques and immunostaining studies were also performed. Positive and negative contrast enhancement of DMNC solutions with increasing Fe concentrations were observed in UTE and T2W imaging, respectively. The relative signal intensities of the DMNC solution containing 2.9 mM Fe were calculated as 3.53 and 0.99 in UTE and T2W imaging, respectively. DMNC uptake into the macrophage cytoplasm was visualized by electron microscopy. Cellular MR imaging of DMNC-treated macrophages revealed relative signals of 3.00 in UTE imaging and 0.98 in T2W imaging. In vivo MR images revealed significant brightening and darkening of plaque areas in the WHHL rabbit 24 h after DMNC injection in UTE and T2W imaging, respectively. Ex vivo MR imaging results agreed with these in vivo MR imaging results. Histological analysis showed that DMNCs were localized to areas of inflammatory vascular wall. Using compensatory UTE and T2W imaging in conjunction with DMNC is an effective approach for the noninvasive in vivo imaging of atherosclerotic plaque.


Renal innervation plays no role in oxygen-dependent control of erythropoietin mRNA levels.
To assess the role of renal innervation in O2-dependent control of erythropoietin (EPO) formation, we have determined EPO mRNA levels in both kidneys of unilaterally denervated rats and sham-operated controls using RNase protection. To investigate whether possible effects of renal nerve input are related to the type of hypoxic stimulus and the degree of stimulation, animals were studied under basal conditions, after exposure to normobaric hypoxia (8% O2, 4 h) or CO (0.1%, 4 h), and after acute hemorrhage (decrease in hematocrit from 40.8 +/- 0.5 to 12.7 +/- 0.5% within 7 h; mean +/- SE, n = 6). Serum EPO levels rose on average 22-, 49-, and 48-fold under the three stimuli and were unaffected by unilateral denervation. Renal EPO mRNA levels in unilaterally denervated animals, when expressed in arbitrary units revealed by comparison with an external standard, were 7.0 +/- 1.5 vs. 6.3 +/- 2.0 (normoxia), 432 +/- 136 vs. 451 +/- 156 (normobaric hypoxia), 971 +/- 93 vs. 930 +/- 120 (CO), and 604 +/- 170 vs. 689 +/- 203 (hemorrhagic anemia) in the intact vs. the denervated kidney (mean +/- SE, n = 3). Furthermore, there was no difference between EPO mRNA levels of either kidney of unilaterally denervated animals and levels in sham-operated controls. We conclude that renal nerve input plays no significant role in the control of the EPO gene under both basal and stimulated conditions.


Novel "Superspreader" Bacteriophages Promote Horizontal Gene Transfer by Transformation.
Bacteriophages infect an estimated 1023 to 1025 bacterial cells each second, many of which carry physiologically relevant plasmids (e.g., those encoding antibiotic resistance). However, even though phage-plasmid interactions occur on a massive scale and have potentially significant evolutionary, ecological, and biomedical implications, plasmid fate upon phage infection and lysis has not been investigated to date. Here we show that a subset of the natural lytic phage population, which we dub "superspreaders," releases substantial amounts of intact, transformable plasmid DNA upon lysis, thereby promoting horizontal gene transfer by transformation. Two novel Escherichia coli phage superspreaders, SUSP1 and SUSP2, liberated four evolutionarily distinct plasmids with equal efficiency, including two close relatives of prominent antibiotic resistance vectors in natural environments. SUSP2 also mediated the extensive lateral transfer of antibiotic resistance in unbiased communities of soil bacteria from Maryland and Wyoming. Furthermore, the addition of SUSP2 to cocultures of kanamycin-resistant E. coli and kanamycin-sensitive Bacillus sp. bacteria resulted in roughly 1,000-fold more kanamycin-resistant Bacillus sp. bacteria than arose in phage-free controls. Unlike many other lytic phages, neither SUSP1 nor SUSP2 encodes homologs to known hydrolytic endonucleases, suggesting a simple potential mechanism underlying the superspreading phenotype. Consistent with this model, the deletion of endonuclease IV and the nucleoid-disrupting protein ndd from coliphage T4, a phage known to extensively degrade chromosomal DNA, significantly increased its ability to promote plasmid transformation. Taken together, our results suggest that phage superspreaders may play key roles in microbial evolution and ecology but should be avoided in phage therapy and other medical applications. Bacteriophages (phages), viruses that infect bacteria, are the planet's most numerous biological entities and kill vast numbers of bacteria in natural environments. Many of these bacteria carry plasmids, extrachromosomal DNA elements that frequently encode antibiotic resistance. However, it is largely unknown whether plasmids are destroyed during phage infection or released intact upon phage lysis, whereupon their encoded resistance could be acquired and manifested by other bacteria (transformation). Because phages are being developed to combat antibiotic-resistant bacteria and because transformation is a principal form of horizontal gene transfer, this question has important implications for biomedicine and microbial evolution alike. Here we report the isolation and characterization of two novel Escherichia coli phages, dubbed "superspreaders," that promote extensive plasmid transformation and efficiently disperse antibiotic resistance genes. Our work suggests that phage superspreaders are not suitable for use in medicine but may help drive bacterial evolution in natural environments.


Interstitial lung disease associated with anti-citrullinated peptide/protein antibody-positive anti-synthetase syndrome.
Little has been reported on the characteristics of interstitial lung disease (ILD) associated with anti-citrullinated peptide/protein antibody (ACPA)-positive anti-synthetase syndrome (ASS). We sought to investigate the clinical, radiologic, and pathologic features of patients with ILD associated with ACPA-positive ASS. Medical records of seven ILD patients with positive results of both ACPA and anti-aminoacyl-tRNA synthetase antibody were retrospectively reviewed. Five patients had clinical symptoms associated with ASS other than ILD. On high-resolution computed tomography (HRCT) analysis, a nonspecific interstitial pneumonia (NSIP) pattern was shown in 3 patients and NSIP with organizing pneumonia (OP) overlap in 2 patients. Coronal slices of these 5 patients showed lower lung disease predominance with traction caudally on major fissures due to lower lobe volume loss. These were features that could commonly be observed in ASS-associated ILD. Pathological findings available for 3 patients showed NSIP. The characteristics of ILD associated with ACPA-positive ASS appear to be similar to those of ILD associated with ASS, but not to rheumatoid arthritis (RA) or ACPA, especially in terms of the radiological findings.


Classical field approach to quantum weak measurements.
By generalizing the quantum weak measurement protocol to the case of quantum fields, we show that weak measurements probe an effective classical background field that describes the average field configuration in the spacetime region between pre- and postselection boundary conditions. The classical field is itself a weak value of the corresponding quantum field operator and satisfies equations of motion that extremize an effective action. Weak measurements perturb this effective action, producing measurable changes to the classical field dynamics. As such, weakly measured effects always correspond to an effective classical field. This general result explains why these effects appear to be robust for pre- and postselected ensembles, and why they can also be measured using classical field techniques that are not weak for individual excitations of the field.


The new chiropractic.
Physical manipulation and manual therapies are thousands of years old. The most popular western world iteration of these therapies is delivered by chiropractors. It can be argued that the collective public health benefit from chiropractic for spinal pain has been very substantial, however as chiropractic has transitioned from craft to profession it has encountered many internally and externally driven machinations that have retarded its progress to a fully accepted allied health profession. This article sets out a ten point plan for a new chiropractic that will achieve full acceptance for this troubled profession. This article is based on a keynote speech known as the FG Roberts Memorial Address delivered on October 10, 2015, in Melbourne, Australia at the Chiropractic & Osteopathic College of Australasia and Chiropractic Australia national conference. The ten point plan consists of the following: improving the pre-professional education of chiropractors, establishing a progressive identity, developing a special interest for the profession, marginalising the nonsensical elements of the profession, being pro-public health, supporting the legitimate organised elements of the profession, improving clinical practice, embracing evidence based practice, supporting research and showing personal leadership. Adherence to this fresh ten point plan will, over time, see the chiropractic profession gain full legitimacy in the allied health field and acceptance by other health providers, policy makers and the public at large.