Impaired endothelium-dependent and independent dilatation of forearm resistance arteries in men with diet-treated non-insulin-dependent diabetes: role of dyslipidaemia.
1. We measured endothelium-dependent and independent dilatation of forearm resistance arteries in 29 men with diet-treated non-insulin-dependent diabetes mellitus and 18 age- and sex-matched control subjects. None of the diabetic patients had hypercholesterolaemia, overt hypertension or microproteinuria. 2. We examined endogenous and exogenous nitric oxide-mediated vasodilatation by measuring forearm blood flow with venous occlusive plethysmography after administration of acetylcholine (7.5 and 15 micrograms/min) and sodium nitroprusside (3 and 10 micrograms/min), respectively, into the brachial artery. NG-monomethyl-L-arginine was also infused to study the inhibition of basal and stimulated release of nitric oxide. 3. The vasodilatory response to acetylcholine, expressed as area under curve, was significantly decreased in the diabetic patients compared with the control subjects (P = 0.019). NG-monomethyl-L-arginine significantly reduced basal (P < 0.001) and acetylcholine-stimulated blood flow (P < 0.02) in both groups. The vasodilatory response (also expressed as area under curve) to sodium nitroprusside was significantly less (P = 0.044) in the diabetic patients than in the control subjects. 4. In the diabetic patients, impaired vasodilatory responses to acetylcholine were significantly correlated with higher serum triacylglycerols (P = 0.048) and lower high-density lipoprotein-cholesterol concentrations (P = 0.007); the association with high-density lipoprotein was independent of age, glycated haemoglobin and blood pressure. Sodium nitroprusside responses were not correlated with lipid and lipoprotein concentrations. 5. We conclude that there is impaired endothelial and smooth muscle cell function in men with diet-treated non-insulin-dependent diabetes mellitus uncomplicated by overt hypertension or microproteinuria. Endothelial dysfunction may be related to diabetic dyslipidaemia and associated metabolic disturbances.


Detection of impaired cerebral autoregulation improves by increasing arterial blood pressure variability.
Although the assessment of dynamic cerebral autoregulation (CA) based on measurements of spontaneous fluctuations in arterial blood pressure (ABP) and cerebral blood flow (CBF) is a convenient and much used method, there remains uncertainty about its reliability. We tested the effects of increasing ABP variability, provoked by a modification of the thigh cuff method, on the ability of the autoregulation index to discriminate between normal and impaired CA, using hypercapnia as a surrogate for dynamic CA impairment. In 30 healthy volunteers, ABP (Finapres) and CBF velocity (CBFV, transcranial Doppler) were recorded at rest and during 5% CO(2) breathing, with and without pseudo-random sequence inflation and deflation of bilateral thigh cuffs. The application of thigh cuffs increased ABP and CBFV variabilities and was not associated with a distortion of the CBFV step response estimates for both normocapnic and hypercapnic conditions (P=0.59 and P=0.96, respectively). Sensitivity and specificity of CA impairment detection were improved with the thigh cuff method, with the area under the receiver-operator curve increasing from 0.746 to 0.859 (P=0.031). We conclude that the new method is a safe, efficient, and appealing alternative to currently existing assessment methods for the investigation of the status of CA.


Surface parameters of ferritic iron-rich Fe-Cr alloy.
Using first-principles density functional theory in the implementation of the exact muffin-tin orbitals method and the coherent potential approximation, we studied the surface energy and the surface stress of the thermodynamically most stable surface facet (100) of the homogeneous disordered body-centred cubic iron-chromium system in the concentration interval up to 20 at.% Cr. For the low-index surface facets of Fe and Cr, the surface energy of Cr is slightly larger than that of Fe, while the surface stress of Cr is considerably smaller than that of Fe. We find that Cr addition to Fe generally increases the surface energy of the Fe-Cr alloy; however, an increase of the bulk amount of Cr also increases the surface stress. As a result of this unexpected trend, the (100) surface of Fe-Cr becomes more stable against reconstruction with increasing Cr concentration. We show that the observed trends are of magnetic origin. In addition to the homogeneous alloy case, we also investigated the impact of surface segregation on both surface parameters.


Evaluation of clinically and physiologically atypical asthma: If it doesn't wheeze it may still be asthma.
Children with asthma-like symptoms may not clinically wheeze. The objectives of this study were to evaluate if children, without physician-documented wheeze, wheeze during bronchial-challenge-testing (BCT), and if measurements of O2Sat and respiratory rate during BCT improve the BCT sensitivity? Seven hundred and twenty-four children, who were referred for suspicion of asthma, performed a BCT. Positive BCT was determined by the provocation concentration (PC) which resulted in a 20% decrease in FEV1 (PC20), (in those who were able to perform spirometry, group B), or (in those unable to perform spirometry, group A) a 50% increase in respiratory rate (PCRR), or a 5% decrease in oxygen-saturation (PCO2-Sat) or appearance of wheezing (PCwheeze). Five hundred and seven BCTs were positive: group A n = 89 age, median (IQR), 3 (2.5-3.7) years (17.6%), were unable to perform spirometry, and group B n = 418 age 10.7 (6.8-15.6) years (82.4%), were able to perform spirometry. Children, without physician-documented wheeze in the total population (groups A plus B), were more likely (65.5%) to have a positive BCT without wheeze compared with those with physician-documented wheeze (41.0%, P < 0.001). In group A, adding PCRR and PCO2-Sat increased BCT sensitivity by 23.6%. Many children in both groups did not wheeze despite reaching BCT endpoints. Children without physician-documented wheeze tended not to wheeze at BCT. This may result in clinical under-diagnosis of asthma if depending on the presence of wheeze. In young children, adding PCRR and PCO2-Sat substantially increases BCT sensitivity and may improve asthma diagnosis.


Retinal mitochondrial DNA mismatch repair in the development of diabetic retinopathy, and its continued progression after termination of hyperglycemia.
Mitochondrial DNA (mtDNA) is damaged in the retina in diabetes, and mitochondria copy numbers are decreased. The displacement-loop (D-loop) of the mtDNA, the region with transcription/replication elements, experiences more damage than other regions of mtDNA. Our aim was to examine the role of DNA mismatch repair (MMR) in mitochondria homeostasis in diabetic retinopathy, and in its continued progression after cessation of hyperglycemia. Effect of hyperglycemia on sequence variants in the D-loop region was investigated in retinal endothelial cells and in the retina from streptozotocin-induced diabetic rats using mismatch-specific surveyor nuclease. The role of MMR machinery in mtDNA damage and mitochondrial respiration was investigated in retinal endothelial cells overexpressing Mlh1, an MMR enzyme mainly associated with mtDNA polymerase gamma, or Msh2 (associated with nuclear polymerase beta). Hyperglycemia increased sequence variants in the D-loop region. While overexpression of Mlh1 in endothelial cells ameliorated glucose-induced increase in D-loop sequence variants, decrease in respiration rate and increase in apoptosis, overexpression of Msh2 did not protect the mitochondria damage. Termination of hyperglycemia failed to reverse decrease in MMR enzymes and increase in D-loop sequence variants. Due to a compromised MMR system, the sequence variants in the D-loop region were not repaired, and that resulted in impaired mtDNA transcription. Mitochondria become dysfunctional, and they continued to be dysfunctional even after hyperglycemia was terminated, contributing to the development, and progression of diabetic retinopathy. Thus, strategies targeting mitochondrial MMR machinery could help maintain mitochondria homeostasis, and inhibit the development of diabetic retinopathy and its continued progression.


A Case with Recurrent Free-Floating Ball Thrombi in Left Atrium.
BACKGROUND A free-floating ball thrombus in the left atrium is a rare clinical condition. However, the diagnosis of this condition has been facilitated by the advent and development of echocardiography and multi-detector row computed tomography (MDCT) and several cases have been reported. CASE REPORT We report a case of a 75-year-old woman who had recurrent giant spherical thrombi in the left atrium. She was diagnosed with chronic atrial fibrillation at 52 years of age. A pacemaker implantation was performed at 54 years of age because of a complete atrioventricular block; and mitral valve replacement was performed for severe mitral regurgitation at 62 years of age. She had a history of cerebral infarction and she was under treatment for chronic heart failure. Despite intensive anticoagulant therapy, she developed ball thrombi in the left atrium three times in six months. During hospitalization for acute myocardial infarction treated with percutaneous catheter intervention, transthoracic echocardiography and computed tomography (CT) revealed a free-floating giant spherical thrombus in the left atrium. She was treated with intensive anticoagulation therapy and the left atrial ball thrombus disappeared; however, two ball thrombi in the left atrium and left atrial appendage recurred after three months. Surgical removal of the thrombi and closure of the left atrial appendage were performed. Unfortunately, a ball thrombus in the left atrium recurred again after a further three months. CONCLUSIONS The present case highlights the difficulty of treating refractory thrombi in the left atrium.


HO-1 overexpression attenuates endotoxin effects on CAT-2 isozymes expression.
l-arginine transport mediated by type-2 cationic amino acid transporter (CAT-2) isozymes is one crucial mechanism that regulates nitric oxide (NO) production via inducible nitric oxide synthase (iNOS). We sought to investigate the effects of heme oxygenase-1 (HO-1) overexpression on CAT-2 isozymes, e.g., CAT-2, CAT-2A, and CAT-2B. Adult male Sprague Dawley rats were allocated to receive lipopolysaccharide (LPS), normal saline, hemin (a HO-1 inducer), tin protoporphyrin (SnPP, a HO-1 inhibitor), LPS plus hemin, or LPS plus hemin plus SnPP. After maintaining for 6 h, rats were sacrificed and the expression and activity of individual enzyme was evaluated. LPS increased HO activity, HO-1 concentration, NO production, l-arginine transport, and concentrations of iNOS, CAT-2, and CAT-2B in rat lungs and kidney. LPS also increased HO activity, HO-1 concentration, NO production, l-arginine transport, and iNOS concentration but decreased CAT-2 and CAT-2B concentrations in rat liver. LPS increased CAT-2A concentration in rat liver but did not affect CAT-2A concentration in rat lungs and kidney. Hemin further increased HO activity and induced HO-1 overexpression in the lungs, kidney, and liver from LPS-treated rats. In addition, the effects of LPS on NO production, l-arginine transport, and concentrations of iNOS and CAT-2 isozymes were significantly attenuated by hemin. SnPP, on the other hand, reversed the effects of hemin. HO-1 overexpression significantly attenuates endotoxin-induced increases in NO production and l-arginine transport. Induction of HO-1 overexpression also significantly attenuates the effects of endotoxin on the expression of iNOS and CAT-2 isozymes in septic rats.


Magnetic fields at neptune.
The National Aeronautics and Space Administration Goddard Space Flight Center-University of Delaware Bartol Research Institute magnetic field experiment on the Voyager 2 spacecraft discovered a strong and complex intrinsic magnetic field of Neptune and an associated magnetosphere and magnetic tail. The detached bow shock wave in the supersonic solar wind flow was detected upstream at 34.9 Neptune radii (R(N)), and the magnetopause boundary was tentatively identified at 26.5 R(N) near the planet-sun line (1 R(N) = 24,765 kilometers). A maximum magnetic field of nearly 10,000 nanoteslas (1 nanotesla = 10(-5) gauss) was observed near closest approach, at a distance of 1.18 R(N). The planetary magnetic field between 4 and 15 R(N) can be well represented by an offset tilted magnetic dipole (OTD), displaced from the center of Neptune by the surprisingly large amount of 0.55 R(N) and inclined by 47 degrees with respect to the rotation axis. The OTD dipole moment is 0.133 gauss-R(N)(3). Within 4 R(N), the magnetic field representation must include localized sources or higher order magnetic multipoles, or both, which are not yet well determined. The obliquity of Neptune and the phase of its rotation at encounter combined serendipitously so that the spacecraft entered the magnetosphere at a time when the polar cusp region was directed almost precisely sunward. As the spacecraft exited the magnetosphere, the magnetic tail appeared to be monopolar, and no crossings of an imbedded magnetic field reversal or plasma neutral sheet were observed. The auroral zones are most likely located far from the rotation poles and may have a complicated geometry. The rings and all the known moons of Neptune are imbedded deep inside the magnetosphere, except for Nereid, which is outside when sunward of the planet. The radiation belts will have a complex structure owing to the absorption of energetic particles by the moons and rings of Neptune and losses associated with the significant changes in the diurnally varying magnetosphere configuration. In an astrophysical context, the magnetic field of Neptune, like that of Uranus, may be described as that of an "oblique" rotator.


Promiscuity and the evolutionary transition to complex societies.
Theory predicts that the evolution of cooperative behaviour is favoured by low levels of promiscuity leading to high within-group relatedness. However, in vertebrates, cooperation often occurs between non-relatives and promiscuity rates are among the highest recorded. Here we resolve this apparent inconsistency with a phylogenetic analysis of 267 bird species, demonstrating that cooperative breeding is associated with low promiscuity; that in cooperative species, helping is more common when promiscuity is low; and that intermediate levels of promiscuity favour kin discrimination. Overall, these results suggest that promiscuity is a unifying feature across taxa in explaining transitions to and from cooperative societies.


Depicting SARS-CoV-2 faecal viral activity in association with gut microbiota composition in patients with COVID-19.
Although severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA was detected in faeces of patients with COVID-19, the activity and infectivity of the virus in the GI tract during disease course is largely unknown. We investigated temporal transcriptional activity of SARS-CoV-2 and its association with longitudinal faecal microbiome alterations in patients with COVID-19. We performed RNA shotgun metagenomics sequencing on serial faecal viral extractions from 15 hospitalised patients with COVID-19. Sequencing coverage of the SARS-CoV-2 genome was quantified. We assessed faecal microbiome composition and microbiome functionality in association with signatures of faecal SARS-CoV-2 infectivity. Seven (46.7%) of 15 patients with COVID-19 had stool positivity for SARS-CoV-2 by viral RNA metagenomic sequencing. Even in the absence of GI manifestations, all seven patients showed strikingly higher coverage (p=0.0261) and density (p=0.0094) of the 3' vs 5' end of SARS-CoV-2 genome in their faecal viral metagenome profile. Faecal viral metagenome of three patients continued to display active viral infection signature (higher 3' vs 5' end coverage) up to 6 days after clearance of SARS-CoV-2 from respiratory samples. Faecal samples with signature of high SARS-CoV-2 infectivity had higher abundances of bacterial species Collinsella aerofaciens, Collinsella tanakaei, Streptococcus infantis, Morganella morganii, and higher functional capacity for nucleotide de novo biosynthesis, amino acid biosynthesis and glycolysis, whereas faecal samples with signature of low-to-none SARS-CoV-2 infectivity had higher abundances of short-chain fatty acid producing bacteria, Parabacteroides merdae, Bacteroides stercoris, Alistipes onderdonkii and Lachnospiraceae bacterium 1_1_57FAA. This pilot study provides evidence for active and prolonged 'quiescent' GI infection even in the absence of GI manifestations and after recovery from respiratory infection of SARS-CoV-2. Gut microbiota of patients with active SARS-CoV-2 GI infection was characterised by enrichment of opportunistic pathogens, loss of salutary bacteria and increased functional capacity for nucleotide and amino acid biosynthesis and carbohydrate metabolism.


Mechanism underlying the DNA-binding preferences of the Vibrio cholerae and vibriophage VP882 VqmA quorum-sensing receptors.
Quorum sensing is a chemical communication process that bacteria use to coordinate group behaviors. In the global pathogen Vibrio cholerae, one quorum-sensing receptor and transcription factor, called VqmA (VqmAVc), activates expression of the vqmR gene encoding the small regulatory RNA VqmR, which represses genes involved in virulence and biofilm formation. Vibriophage VP882 encodes a VqmA homolog called VqmAPhage that activates transcription of the phage gene qtip, and Qtip launches the phage lytic program. Curiously, VqmAPhage can activate vqmR expression but VqmAVc cannot activate expression of qtip. Here, we investigate the mechanism underlying this asymmetry. We find that promoter selectivity is driven by each VqmA DNA-binding domain and key DNA sequences in the vqmR and qtip promoters are required to maintain specificity. A protein sequence-guided mutagenesis approach revealed that the residue E194 of VqmAPhage and A192, the equivalent residue in VqmAVc, in the helix-turn-helix motifs contribute to promoter-binding specificity. A genetic screen to identify VqmAPhage mutants that are incapable of binding the qtip promoter but maintain binding to the vqmR promoter delivered additional VqmAPhage residues located immediately C-terminal to the helix-turn-helix motif as required for binding the qtip promoter. Surprisingly, these residues are conserved between VqmAPhage and VqmAVc. A second, targeted genetic screen revealed a region located in the VqmAVc DNA-binding domain that is necessary to prevent VqmAVc from binding the qtip promoter, thus restricting DNA binding to the vqmR promoter. We propose that the VqmAVc helix-turn-helix motif and the C-terminal flanking residues function together to prohibit VqmAVc from binding the qtip promoter.


The histone chaperone Nap1 promotes nucleosome assembly by eliminating nonnucleosomal histone DNA interactions.
The organization of the eukaryotic genome into nucleosomes dramatically affects the regulation of gene expression. The delicate balance between transcription and DNA compaction relies heavily on nucleosome dynamics. Surprisingly, little is known about the free energy required to assemble these large macromolecular complexes and maintain them under physiological conditions. Here, we describe the thermodynamic parameters that drive nucleosome formation in vitro. To demonstrate the versatility of our approach, we test the effect of DNA sequence and H3K56 acetylation on nucleosome thermodynamics. Furthermore, our studies reveal the mechanism of action of the histone chaperone nucleosome assembly protein 1 (Nap1). We present evidence for a paradigm in which nucleosome assembly requires the elimination of competing, nonnucleosomal histone-DNA interactions by Nap1. This observation is confirmed in vivo, wherein deletion of the NAP1 gene in yeast results in a significant increase in atypical histone-DNA complexes, as well as in deregulated transcription activation and repression.


Core/Whole Genome Multilocus Sequence Typing and Core Genome SNP-Based Typing of OXA-48-Producing Klebsiella pneumoniae Clinical Isolates From Spain.
Whole-genome sequencing (WGS)-based typing methods have emerged as promising and highly discriminative epidemiological tools. In this study, we combined gene-by-gene allele calling and core genome single nucleotide polymorphism (cgSNP) approaches to investigate the genetic relatedness of a well-characterized collection of OXA-48-producing Klebsiella pneumoniae isolates. We included isolates from the predominant sequence type ST405 (n = 31) OXA-48-producing K. pneumoniae clone and isolates from ST101 (n = 3), ST14 (n = 1), ST17 (n = 1), and ST1233 (n = 1), obtained from eight Catalan hospitals. Core-genome multilocus sequence typing (cgMLST) schemes from Institut Pasteur's BIGSdb-Kp (634 genes) and SeqSphere+ (2,365 genes), and a SeqSphere+ whole-genome MLST (wgMLST) scheme (4,891 genes) were used. Allele differences or allelic mismatches and the genetic distance, as the proportion of allele differences, were used to interpret the results from a gene-by-gene approach, whereas the number of SNPs was used for the cgSNP analysis. We observed between 0-10 and 0-14 allele differences among the predominant ST405 using cgMLST and wgMLST from SeqSphere+, respectively, and <2 allelic mismatches when using Institut Pasteur's BIGSdb-Kp cgMLST scheme. For ST101, we observed 14 and 54 allele differences when using cgMLST and wgMLST SeqSphere+, respectively, and 2-5 allelic mismatches for BIGSdb-Kp cgMLST. A low genetic distance (<0.0035, a previously established threshold for epidemiological link) was generally in concordance with a low number of allele differences (<8) when using the SeqSphere+ cgMLST scheme. The cgSNP analysis showed 6-29 SNPs in isolates with identical allelic SeqSphere+ cgMLST profiles and 16-61 cgSNPs among ST405 isolates. Furthermore, comparison of WGS-based typing results with previously obtained MLST and pulsed-field gel electrophoresis (PFGE) data showed some differences, demonstrating the different molecular principles underlying these techniques. In conclusion, the use of the different WGS-based typing methods that were used to elucidate the genetic relatedness of clonal OXA-48-producing K. pneumoniae all led to the same conclusions. Furthermore, threshold parameters in WGS-based typing methods should be applied with caution and should be used in combination with clinical epidemiological data and population and species characteristics.


Alcohol's contribution to compromised immunity.
Alcoholics frequently suffer from infectious diseases and have increased rates of some cancers, indicating that alcohol impairs the immune system, which protects the body against this type of damage. Alcohol interferes with the functions of many of the cells and molecules that are part of the immune system. For example, alcohol inhibits the functions of the cells that ingest and destroy invading microorganisms (i.e., neutrophils, monocytes, and macrophages). Both acute and chronic alcohol exposure also alter the production of signaling molecules that help coordinate the immune response (i.e., cytokines). Finally, alcohol adversely affects the functions of the cells that mediate the immune response against specific microorganisms and long-term immunity (i.e., T cells and B cells). As a result, alcoholics have an increased susceptibility to diseases caused by bacterial infections, such as tuberculosis and pneumonia. Alcoholics also may be more susceptible to infections from the virus that causes AIDS. In addition, alcohol intoxication can exacerbate the immune suppression that occurs after traumatic injuries.


Studies on the cornea. II. The fine structure of Descement's membrane.
Descemet's membrane, previously thought to be "structureless," has been found to be characterized by a fine structure of great regularity. Sections perpendicular to the plane of the membrane surface appeared to be cross-striated, with narrow dark bands separated by wide light bands traversed by fine filaments. Tangential sections showed a two-dimensional array of dark nodes and thin internodal filaments which connected each node with the six others around it to form a hexagonal figure. The average distance between nodes, and between the dark bands in transverse sections, was about 1070 A; the width of the nodes was about 270 A; and the width of the connecting filaments was less than 100 A. This pattern has been found in all species of Descemet's membrane so far examined, although it appeared to be better developed in some forms than in others. So far as is known, it has not been observed in any other type of tissue. The differentiation of Descemet's membrane has been followed in the chick embryo. It appeared late and developed slowly but showed the characteristic fine structure from the time it could be clearly identified. The evidence for placing Descemet's membrane in the collagen class of proteins is strongly supplemented by the results obtained from x-ray diffraction examination by Rougvie and Bear.


A grounded theory approach to understand the process of decision making on fertility control methods in urban society of Mashhad, Iran.
More than 30% of pregnancies in Iran are unintended and most of them happen among the women who use various contraceptive methods. Results of Integrated Monitoring and Evaluation System (IMES) showed that the rate of innovative contraceptive use in Mashhad has been 41.5%-57% in different urban areas. This study was conducted to explore the process of making decision toward using family planning methods in women of reproductive age in urban society of Mashhad, Iran. In this grounded theory study, semi-structured interviews were conducted with 45 purposefully selected participants including 28 women and 17 key informants including family health providers and managers, and participants' mothers and husbands, who lived in urban society of Mashhad, Iran, in 2011-2012. Participants' recruitment continued until data saturation occurred. Data were analyzed using Strauss and Corbin's mode of analysis through constant comparative method, applying levels of open, axial, and selective coding with MAXqda software. Study rigor was confirmed through prolonged engagement, member check, expert debriefing, and thick description of the data. The core category of "caring the comprehensive health of my family," which emerged from the data, described the process of couples' decision making toward using family planning methods in this study. Other developed categories which were presented into a theoretical scheme consisted of 1) shaping the ideas of fertility control, 2) developing cognition about the fertility control methods, 3) appraising available choices and choosing the most appropriate one, 4) managing the course of using methods, and 5) realizing the fertility intentions. It is important that family planning providers understand the motivations, perceptions, and knowledge of women about contraceptive methods in their contextual situation, which illustrates their mode of interaction in the arenas of family planning decision making.


Evaluation of the presentation of systemic onset juvenile rheumatoid arthritis: data from the Pennsylvania Systemic Onset Juvenile Arthritis Registry (PASOJAR).
To characterize the initial clinical and laboratory features of patients with systemic onset juvenile rheumatoid arthritis (soJRA) through a Web-based registry. Patients diagnosed with soJRA in the last 15 years at 3 medical centers in Pennsylvania were identified. Data were collected retrospectively using a Web-based interface in compliance with patient privacy standards. Inferential statistics were used to compare features of patients with and without macrophage activation syndrome. We identified 136 patients; 88% of patients presented with arthritis (8% mono-, 45% oligo-, 47% polyarticular). The most common joints involved were the knee (68% of patients with arthritis), wrist (68%), and ankle (57%). The International League of Associations for Rheumatology criteria for systemic juvenile idiopathic arthritis (SJIA) identified only 30% of patients at presentation. We successfully characterized the presenting features of a relatively rare disease, soJRA, through the use of a Web-based registry. Current classification criteria for SJIA may not be particularly sensitive for diagnosis at presentation.


HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENES1 is required for circadian periodicity through the promotion of nucleo-cytoplasmic mRNA export in Arabidopsis.
Cold acclimation has been shown to be attenuated by the degradation of the INDUCER OF CBF EXPRESSION1 protein by the E3 ubiquitin ligase HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENES1 (HOS1). However, recent work has suggested that HOS1 may have a wider range of roles in plants than previously appreciated. Here, we show that hos1 mutants are affected in circadian clock function, exhibiting a long-period phenotype in a wide range of temperature and light environments. We demonstrate that hos1 mutants accumulate polyadenylated mRNA in the nucleus and that the circadian defect in hos1 is shared by multiple mutants with aberrant mRNA export, but not in a mutant attenuated in nucleo-cytoplasmic transport of microRNAs. As revealed by RNA sequencing, hos1 exhibits gross changes to the transcriptome with genes in multiple functional categories being affected. In addition, we show that hos1 and other previously described mutants with altered mRNA export affect cold signaling in a similar manner. Our data support a model in which altered mRNA export is important for the manifestation of hos1 circadian clock defects and suggest that HOS1 may indirectly affect cold signaling through disruption of the circadian clock.


Transposable elements: from DNA parasites to architects of metazoan evolution.
One of the most unexpected insights that followed from the completion of the human genome a decade ago was that more than half of our DNA is derived from transposable elements (TEs). Due to advances in high throughput sequencing technologies it is now clear that TEs comprise the largest molecular class within most metazoan genomes. TEs, once categorised as "junk DNA", are now known to influence genomic structure and function by increasing the coding and non-coding genetic repertoire of the host. In this way TEs are key elements that stimulate the evolution of metazoan genomes. This review highlights several lines of TE research including the horizontal transfer of TEs through host-parasite interactions, the vertical maintenance of TEs over long periods of evolutionary time, and the direct role that TEs have played in generating morphological novelty.


The CAN1 locus of Saccharomyces cerevisiae: fine-structure analysis and forward mutation rates.
A system of strains and growth media was developed to allow efficient detection of forward mutation, reversion, complementation, and suppression at the canavanine-resistance (CAN1) locus of Saccharomyces cerevisiae. Genetic fine-structure analysis revealed that the map length is at least 40, and possibly as much as 60 X-ray map units; this is the longest gene map yet reported in S. cerevisiae. Allelic complementation was not observed, despite testing of a large number of allele pairs, and alleles suppressible by the ochre suppressor SUP11 were absent from a sample of 48 spontaneous mutants and occurred infrequently (7%) among a sample of ultraviolet-induced mutants. Infrequent mutant types included canavanine-resistant mutants capable of arginine uptake and alleles thought to represent deletions or inversions. In contrast to previous reports in the literature, the spontaneous forward mutation rate at CAN1 did not increase during meiosis.


Home improvement and system-based health promotion for sustainable prevention of Chagas disease: A qualitative study.
Human transmission of Chagas disease (CD) most commonly occurs in domiciliary spaces where triatomines remain hidden to feed on blood sources during inhabitants' sleep. Similar to other neglected tropical diseases (NTDs), sustainable control of CD requires attention to the structural conditions of life of populations at risk, in this case, the conditions of their living environments. Considering socio-cultural and political dynamics involved in dwellings' construction, this study aimed to explore social factors that contribute or limit sustainability of CD's prevention models focused on home improvement. Using Healthy Homes for Healthy Living (HHHL)-a health promotion strategy focused on improvement of living environments and system-based health promotion-as a reference, a qualitative study was conducted. Research participants were selected from three rural communities of a CD endemic region in southern Ecuador involved in HHHL's refurbishment and reconstruction interventions between 2013 and 2016. Folowing an ethnographic approach, data were collected through interviews, participant observation, informal conversations and document analysis. Our results indicate that the HHHL model addressed risk factors for CD at the household level, while simultaneously promoting wellbeing at emotional, economic and social levels in local communities. We argue that sustainability of the CD prevention model proposed by HHHL is enhanced by the confluence of three factors: systemic improvement of families' quality of life, perceived usefulness of control measures, and flexibility to adapt to emerging dynamics of the context. HHHL's proposed home improvement, facilitated through system-based rather than disease specific health promotion processes, enhances agency in populations at risk and facilitates community partnerships forged around CD prevention. Although an independent analysis of cost-effectiveness is recommended, structural poverty experienced by local families is still the most important factor to consider when evaluating the sustainability and scalability of this model.


Epidemiology of inflammatory bowel diseases in childhood.
Inflammatory Bowel Disease (IBD) is common in most industrialised countries and childhood IBD accounts for nearly 30% of total cases. Various studies, mostly from Europe and USA have reported epidemiological characteristics of childhood IBD. The incidence figures vary greatly from region to region and within a region over time. Almost all reported studies have documented an increase in the incidence, mainly of Crohn's disease over the last few decades. The reasons for the increase are not clear but epidemiological observations have led to many postulates. Incidence in developing countries is perceived to be low, but limited data suggest that it may not be as uncommon as previously thought. IBD can occur at any age but is rare in infancy. Among childhood IBD, early onset IBD appears to be different epidemiologically and is characterised by predominance of colonic involvement and high positive family history. It has become apparent that only about 25% of childhood Crohn's disease presents with classical triad of abdominal pain, diarrhoea and weight loss. Pediatricians should be aware of atypical manifestations and should maintain high index of suspicion. Though epidemiological studies of childhood IBD done so far have contributed towards understanding of IBD, they have differed in study design, population, time period, age group and case definitions. Unfortunately there are no uniform, clear diagnostic criteria which are evidence based. To address this problem, recently the IBD working group of European Society of Pediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN) has published "The Porto Criteria" which details a consensus based diagnostic criteria for the diagnosis of childhood IBD. This should bring uniformity in ascertainment of newly diagnosed IBD cases. An European multicentre prospective database has also been established to facilitate future epidemiological studies.


Langerhans cell homeostasis and turnover after nonmyeloablative and myeloablative allogeneic hematopoietic cell transplantation.
Langerhans cells (LCs) are self-renewing epidermal myeloid cells that can migrate and mature into dendritic cells. Recipient LCs that survive cytotoxic therapy given in preparation for allogeneic hematopoietic cell transplantation may prime donor T cells to mediate cutaneous graft-versus-host disease (GVHD). This possible association, however, has not been investigated in the setting of nonmyeloablative allografting. We prospectively studied the kinetics of LC-chimerism after sex-mismatched allogeneic hematopoietic cell transplantation with nonmyeloablative (n=23) or myeloablative (n=25) conditioning. Combined XY-FISH and Langerin-staining was used to assess donor LC-chimerism in skin biopsies obtained on days 28, 56, and 84 after transplant. The degree of donor LC-chimerism was correlated with the development of skin GVHD. We observed significantly delayed donor LC-engraftment after nonmyeloablative transplantation compared with other hematopoietic compartments and compared with LC-engraftment after myeloablative conditioning. In most recipients of nonmyeloablative transplants, recipient LCs proliferated in situ, recruitment of donor-LCs was delayed by two months, and full donor LC-chimerism was only reached by day 84 after transplant. Although persistence of host LCs on day-28 after transplant was not predictive for acute or chronic skin GVHD, the recruitment of donor-derived LCs was associated with nonspecific inflammatory infiltrates (P=0.009). These results show that LCs can self-renew locally but are replaced by circulating precursors even after minimally toxic nonmyeloablative transplant conditioning. Cutaneous inflammation accompanies donor LC-engraftment, but differences in LC conversion-kinetics do not predict clinical or histopathological GVHD.


Acute cauda equina syndrome secondary to a lumbar synovial cyst.
Cauda equina syndrome is a well described state of neurologic compromise due to lumbosacral root compression. In most cases, it is due to a herniated disc, tumor, infection, or hematoma. We report a case of rapid lumbar synovial cyst expansion leading to acute cauda equina syndrome and compare it to similar cases in the literature. The patient is a 49-year-old woman with a history of chronic low back pain who developed cauda equina syndrome. Serial lumbar magnetic resonance imaging studies demonstrated a significant increase in the size of a lumbar synovial cyst over a 2 week interval. After an unsuccessful attempt to relieve her acute symptoms with computed tomography-guided cyst aspiration, an L4-5 posterior spinal decompression with excision of the synovial cyst was performed. Postoperatively the patient's perineal numbness, bladder incontinence, and associated pain complaints resolved. The only residual symptom at one month follow-up was continued numbness in the right lower limb in an L5 distribution. This report adds to 6 other well described similar cases found in the literature by illustrating several important points. First, a lumbar synovial cyst is a rare but possible cause of acute cauda equina syndrome. Second, magnetic resonance imaging is the test of choice to diagnose and characterize lumbar synovial cysts; serial imaging can detect fluctuations in cyst size. Third, percutaneous treatment of lumbar synovial cysts is variable in efficacy and proved to be unsuccessful in our patient. Finally, surgical management has shown high success rates for symptomatic cysts. Specifically, in the setting of acute cauda equina syndrome secondary to a lumbar synovial cyst, urgent surgical decompression has led to resolution of neurologic symptoms in most reported cases. A lumbar synovial cyst is an uncommon cause of acute cauda equina syndrome. Prompt diagnosis and treatment may lead to reduced morbidity associated with this condition.


Molecular phylogenetics and historical biogeography of Rhinolophus bats.
The phylogenetic relationships within the horseshoe bats (genus Rhinolophus) are poorly resolved, particularly at deeper levels within the tree. We present a better-resolved phylogenetic hypothesis for 30 rhinolophid species based on parsimony and Bayesian analyses of the mitochondrial cytochrome b gene and three nuclear introns (TG, THY and PRKC1). Strong support was found for the existence of two geographic clades within the monophyletic Rhinolophidae: an African group and an Oriental assemblage. The relaxed Bayesian clock method indicated that the two rhinolophid clades diverged approximately 35 million years ago and results from Dispersal Vicariance (DIVA) analysis suggest that the horseshoe bats arose in Asia and subsequently dispersed into Europe and Africa.


Effect of compensation method on the behavior of primary care physicians in managed care organizations: evidence from interviews with physicians and medical leaders in Washington State.
The perceived relationship between primary care physician compensation and utilization of medical services in medical groups affiliated with one or more among six managed care organizations in the state of Washington was examined. Representatives from 67 medical group practices completed a survey designed to determine the organizational arrangements and norms that influence primary care practice and to provide information on how groups translate the payments they receive from health plans into individual physician compensation. Semistructured interviews with 72 individual key informants from 31 of the 67 groups were conducted to ascertain how compensation method affects physician practice. A team of raters read the transcripts and identified key themes that emerged from the interviews. The themes generated from the key informant interviews fell into three broad categories. The first was self-selection and satisfaction. Compensation method was a key factor for physicians in deciding where to practice. Physicians' satisfaction with compensation method was high in part because they chose compensation methods that fit with their practice styles and lifestyles. Second, compensation drives production. Physician production, particularly the number of patients seen, was believed to be strongly influenced by compensation method, whereas utilization of ancillary services, patient outcomes, and satisfaction are seen as much less likely to be influenced. The third theme involved future changes in compensation methods. Medical leaders, administrators, and primary care physicians in several groups indicated that they expected changes in the current compensation methods in the near future in the direction of incentive-based methods. The responses revealed in interviews with physicians and administrative leaders underscored the critical role compensation arrangements play in driving physician satisfaction and behavior.


Robust detection and verification of linear relationships to generate metabolic networks using estimates of technical errors.
The size and magnitude of the metabolome, the ratio between individual metabolites and the response of metabolic networks is controlled by multiple cellular factors. A tight control over metabolite ratios will be reflected by a linear relationship of pairs of metabolite due to the flexibility of metabolic pathways. Hence, unbiased detection and validation of linear metabolic variance can be interpreted in terms of biological control. For robust analyses, criteria for rejecting or accepting linearities need to be developed despite technical measurement errors. The entirety of all pair wise linear metabolic relationships then yields insights into the network of cellular regulation. The Bayesian law was applied for detecting linearities that are validated by explaining the residues by the degree of technical measurement errors. Test statistics were developed and the algorithm was tested on simulated data using 3-150 samples and 0-100% technical error. Under the null hypothesis of the existence of a linear relationship, type I errors remained below 5% for data sets consisting of more than four samples, whereas the type II error rate quickly raised with increasing technical errors. Conversely, a filter was developed to balance the error rates in the opposite direction. A minimum of 20 biological replicates is recommended if technical errors remain below 20% relative standard deviation and if thresholds for false error rates are acceptable at less than 5%. The algorithm was proven to be robust against outliers, unlike Pearson's correlations. The algorithm facilitates finding linear relationships in complex datasets, which is radically different from estimating linearity parameters from given linear relationships. Without filter, it provides high sensitivity and fair specificity. If the filter is activated, high specificity but only fair sensitivity is yielded. Total error rates are more favorable with deactivated filters, and hence, metabolomic networks should be generated without the filter. In addition, Bayesian likelihoods facilitate the detection of multiple linear dependencies between two variables. This property of the algorithm enables its use as a discovery tool and to generate novel hypotheses of the existence of otherwise hidden biological factors.


Spatial instabilities, all-optical limiting, and thresholding in nonlinear distributed-feedback devices.
Solutions of coupled-mode equations describing intensity-induced Bragg coupling of counterpropagating modes in a nonlinear distributed-feedback structure may become spatially unstable. Instability leads to nonpassing plateaus in the intensity transmission characteristics. This induces strong optical limiting action in the transmission mode and lower thresholding in the reflection mode.


Extreme attributions predict the course of bipolar depression: results from the STEP-BD randomized controlled trial of psychosocial treatment.
Little is known about predictors of recovery from bipolar depression or moderators of treatment response. In the present study, we investigated attributional style (a cognitive pattern of explaining the causes of life events) as a predictor of recovery from episodes of bipolar depression and as a moderator of response to psychotherapy for bipolar depression. 106 depressed outpatients with DSM-IV bipolar I or II disorder who were enrolled in the Systematic Treatment Enhancement Program for Bipolar Disorder were randomly assigned to intensive psychotherapy for depression (n = 62) or to collaborative care (n = 44), a minimal psychoeducational intervention. The primary outcome was recovery status at each study visit as measured by the Clinical Monitoring Form. Attributional style was measured at baseline using the Attributional Style Questionnaire. Data were collected between 1998 and 2005. All analyses were by intention to treat. Extreme attributions predicted a lower likelihood of recovery (P < .01; OR = 0.93; 95% CI, 0.88-0.98) and longer time until recovery (P < .01; OR = 0.96; 95% CI, 0.93-0.99), independent of the effects of initial depression severity. Among individuals with more pessimistic attributional styles, higher initial depression severity predicted a lower likelihood of recovery (P = .01; OR = 0.64; 95% CI, 0.45-0.91) and longer time until recovery (P < .001; OR = 0.76; 95% CI, 0.66-0.88). There was no difference in recovery rates between intensive psychotherapy and collaborative care (OR = 0.90; 95% CI, 0.40-2.01) in the full sample. These results suggest that extreme, rigid attributions may be associated with a more severe course of depression and that evaluating attributional style may help clinicians to identify patients who are at risk for experiencing a more severe course of depression. ClinicalTrials.gov identifier: NCT00012558.


Comparing the effects of Calendula officinalis and clotrimazole on vaginal Candidiasis: A randomized controlled trial.
This triple-blind trial examined the effects of Calendula officinalis vaginal cream on the treatment of vaginal Candidiasis (primary outcome) and sexual function (secondary outcome). Married women aged 18-45 years with vaginal Candidiasis (n = 150) were recruited from April to October 2014 and randomized into Calendula and clotrimazole groups, using 5-g vaginal cream every night for seven nights. Clinical and laboratory assessments were conducted at 10-15 and 30-35 days after intervention and the female sexual function index was assessed at 30-35 days. Six women were lost to follow-up. The frequency of testing negative for Candidiasis in the Calendula group was significantly lower at the first (49% vs. 74%; odds ratio (OR) 0.32; 95% confidence interval (CI) 0.16-0.67) but higher at the second (77% vs. 34%; OR 3.1; 95% CI 1.5-6.2) follow-up compared to the clotrimazole group. The frequency of most signs and symptoms were almost equal in the two groups at the first follow-up, but were significantly lower in the Calendula group at the second follow-up. Sexual function had almost equal significant improvement in both groups. Calendula vaginal cream appears to have been effective in the treatment of vaginal Candidiasis and to have a delayed but greater long-term effect compared to clotrimazole.


prlA suppressors in Escherichia coli relieve the proton electrochemical gradient dependency of translocation of wild-type precursors.
The SecY protein of Escherichia coli is an integral membrane component of the protein export apparatus. Suppressor mutations in the secY gene (prlA alleles) have been isolated that restore the secretion of precursor proteins with defective signal sequences. These mutations have never been shown to affect the translocation of wild-type precursor proteins. Here, we report that prlA suppressor mutations relieve the proton-motive force (pmf) dependency of the translocation of wild-type precursors, both in vivo and in vitro. Furthermore, the proton-motive force dependency of the translocation of a precursor with a stably folded domain in the mature region was suppressed by prlA mutations in vitro. These data show that prlA mutations cause a general relaxation of the export apparatus rather than a specific change that results in bypassing of the recognition of the signal sequence. In addition, these results are indicative for a mechanism in which the proton-motive force stimulates translocation by altering the conformation of the translocon.


Fabrication of ordered bulk heterojunction organic photovoltaic cells using nanopatterning and electrohydrodynamic spray deposition methods.
Organic photovoltaic cells with an ordered heterojunction (OHJ) active layer are expected to show increased performance. In the study described here, OHJ cells were fabricated using a combination of nanoimprinting and electrohydrodynamic (EHD) spray deposition methods. After an electron donor material was nanoimprinted with a PDMS stamp (valley width: 230 nm, period: 590 nm) duplicated from a Si nanomold, an electron acceptor material was deposited onto the nanoimprinted donor layer using an EHD spray deposition method. The donor-acceptor interface layer was observed by obtaining cross-sectional images with a focused ion beam (FIB) microscope. The photocurrent generation performance of the OHJ cells was evaluated with the current density-voltage curve under air mass (AM) 1.5 conditions. It was found that the surface morphology of the electron acceptor layer affected the current and voltage outputs of the photovoltaic cells. When an electron acceptor layer with a smooth thin (250 nm above the valley of the electron donor layer) surface morphology was obtained, power conversion efficiency was as high as 0.55%. The electrohydrodynamic spray deposition method used to produce OHJ photovoltaic cells provides a means for the adoption of large area, high throughput processes.


Mindfulness-based stress reduction for solid organ transplant recipients: a randomized controlled trial.
Patients who have received solid organ transplants continue to experience a myriad of complex symptoms related to their underlying disease and to chronic immunosuppression that reduce the quality of life. Beneficial nonpharmacologic therapies to address these symptoms have not been established in the transplant population. Assess the efficacy of mindfulness-based stress reduction (MBSR) in reducing symptoms of anxiety, depression, and poor sleep in transplant patients. Controlled trial with a two-staged randomization. Recipients of kidney, kidney/pancreas, liver, heart, or lung transplants were randomized to MBSR (n=72) or health education (n=66) initially or after serving in a waitlist. Mean age was 54 years (range 21-75); 55% were men, and 91% were white. MBSR, a mindfulness meditation training program consisting of eight weekly 2.5-hour classes; health education, a peer-led active control. Anxiety (State-Trait Anxiety Inventory), depression (Center for Epidemiologic Studies Depression Scale), and sleep quality (Pittsburgh Sleep Quality Index) scales assessed by self-report at baseline, 8 weeks, 6 months, and 1 year. Benefits of MBSR were above and beyond those afforded by the active control. MBSR reduced anxiety and sleep symptoms (P < .02), with medium treatment effects (.51 and .56) at 1 year compared to health education in intention-to-treat analyses. Within the MBSR group, anxiety, depression, and sleep symptoms decreased and quality-of-life measures improved by 8 weeks (P < .01, all), and benefits were retained at 1 year (P < .05, all). Initial symptom reductions in the health education group were smaller and not sustained. Comparisons to the waitlist confirmed the impact of MBSR on both symptoms and quality of life, whereas health education improvements were limited to quality-of-life ratings. MBSR reduced distressing symptoms of anxiety, depression, and poor sleep and improved quality of life. Benefits were sustained over 1 year. A health education program provided fewer benefits, and effects were not as durable. MBSR is a relatively inexpensive, safe, and effective community-based intervention.


Ex vivo hepatic gene transfer in mouse using a defective herpes simplex virus-1 vector.
A defective amplicon herpes simplex virus-1 (HSV-1) vector, HSVlac, was used to transfer an E. coli lacZ reporter gene into primary hepatocytes. The lacZ gene was driven by the HSV immediate early (IE) 4/5 promoter. Use of the HSVlac vector resulted in highly efficient gene transfer. Because difficulties in culturing primary hepatocytes impose limitations in ex vivo gene therapy, we sought to determine whether use of the HSVlac vector could simplify gene transfer. Therefore, we incubated HSVlac with primary hepatocytes in suspension and found that the lacZ gene was still transferred with great rapidity and efficiency. To examine lacZ expression in transduced hepatocytes in vivo, we used a mouse hepatocyte transplantation system. In congeneic recipients of primary hepatocytes transduced with HSVlac in suspension, the lacZ gene was expressed in liver and spleen up to 2 weeks. However, survival of transplanted hepatocytes, as well as persistence of HSVlac genome in recipient organs, was demonstrated for up to an 11-week duration of the experiment. These findings suggest that in vivo regulation of the HSV IE4/5 promoter was responsible for the short-term expression of lacZ, which should be overcome by the use of liver-specific promoters. Therefore, our results indicate the feasibility of hepatic gene transfer with a defective HSV-1 vector.


Carbon benefits of wolfberry plantation on secondary saline land in Jingtai oasis, Gansu--A case study on application of the CBP model.
The largest global source of anthropogenic CO2 emissions comes from the burning of fossil fuel and approximately 30% of total net emissions come from land use and land use change. Forestation and reforestation are regarded worldwide as effective options of sequestering carbon to mitigate climate change with relatively low costs compared with industrial greenhouse gas (GHG) emission reduction efforts. Cash trees with a steady augmentation in size are recognized as a multiple-beneficial solution to climate change in China. The reporting of C changes and GHG emissions for sustainable land management (SLM) practices such as afforestation is required for a variety of reasons, such as devising land management options and making policy. The Carbon Benefit Project (CBP) Simple Assessment Tool was employed to estimate changes in soil organic carbon (SOC) stocks and GHG emissions for wolfberry (Lycium barbarum L.) planting on secondary salinized land over a 10 year period (2004-2014) in the Jingtai oasis in Gansu with salinized barren land as baseline scenario. Results show that wolfberry plantation, an intensively managed ecosystem, served as a carbon sink with a large potential for climate change mitigation, a restorative practice for saline land and income stream generator for farmers in soil salinized regions in Gansu province. However, an increase in wolfberry production, driven by economic demands, would bring environmental pressures associated with the use of N fertilizer and irrigation. With an understanding of all of the components of an ecosystem and their interconnections using the Drivers-Pressures-State-Impact-Response (DPSIR) framework there comes a need for strategies to respond to them such as capacity building, judicious irrigation and institutional strengthening. Cost benefit analysis (CBA) suggests that wolfberry cultivation was economically profitable and socially beneficial and thus well-accepted locally in the context of carbon sequestration. This study has important implications for Gansu as it helps to understand the role cash trees can play in carbon emission reductions. Such information is necessary in devising management options for sustainable land management (SLM).


WE-G-BRB-01: Per-Frame Analysis of MatriXX Data.
One of the most widely used IMRT QA devices (MatriXX) is an array of ionization chambers which are periodically read during plan delivery. Although the ionization chambers are not expected to exhibit strong angular dependence, the measured dose distribution is often found to significantly differ from the planned dose distribution. We identify the origin of all factors that affect the measurement accuracy of the MatriXX and develop a per-frame post-processing strategy that reduces their impact on the passing rate of IMRT and VMAT plans. We developed software that reads the dose frame sequence recorded by the MatriXX and applies a number of correction factors to each frame. Angular correction factors are computed as ratios of measured dose at the isocenter of the phantom and planned dose at the same location for all clinically used photon energies. For every clinical case, the recorded movie file is read and the dose for every frame is corrected according to the angle of the beam. In addition, the background evolution is tracked in the 'beam-off' frames which are subsequently subtracted from the 'beam-on' frames according to a predictive model. Machine output correction is also implemented, which significantly improves the absolute dose measurements. The IMRT effective plane of measurement of the MatriXX was identified and found not to coincide with the isocentric plane. The clinical passing rates are significantly improved when the per-frame analysis software was introduced in our IMRT QA procedure. For a group of 800 patients with no corrections the average passing rate was 93.6%, while for the first 300 cases with per frame corrections the average passing rate was 97.3%. We identify all factors that impact the measurement accuracy of the MatriXX (angular effects, background evolution, machine output, plane of measurement) and propose a strategy for their elimination.


Sustained prognostic implications of newly detected glucose abnormalities in patients with acute myocardial infarction: long-term follow-up of the Glucose Tolerance in Patients with Acute Myocardial Infarction cohort.
To investigate long-term prognostic importance of newly discovered glucose disturbances in patients with acute myocardial infarction (AMI). During 1998-2001, consecutive patients with AMI (n = 167) and healthy controls (n = 184) with no previously known diabetes were investigated with an oral glucose tolerance test (OGTT). Patients and controls were separately followed up for cardiovascular events (first of cardiovascular mortality/AMI/stroke/heart failure) during a decade. In all, 68% of the patients and 35% of the controls had newly detected abnormal glucose tolerance (AGT). Cardiovascular event (n = 72, p = 0.0019) and cardiovascular mortality (n = 31, p = 0.031) were more frequent in patients with newly detected AGT. Regarding patients, a Cox proportional-hazard regression analysis identified AGT (hazard ratio (HR): 2.30; 95% confidence interval (CI): 1.24-4.25; p = 0.008) and previous AMI (HR: 2.39; CI: 1.31-4.35; p = 0.004) as prognostically important. An OGTT at discharge after AMI disclosed a high proportion of patients with previously unknown AGT which had a significant and independent association with long-term prognosis.


Identification of Vimentin as a Potential Therapeutic Target against HIV Infection.
A combination of antiviral drugs known as antiretroviral therapy (ART) has shown effectiveness against the human immunodeficiency virus (HIV). ART has markedly decreased mortality and morbidity among HIV-infected patients, having even reduced HIV transmission. However, an important current disadvantage, resistance development, remains to be solved. Hope is focused on developing drugs against cellular targets. This strategy is expected to prevent the emergence of viral resistance. In this study, using a comparative proteomic approach in MT4 cells treated with an anti-HIV leukocyte extract, we identified vimentin, a molecule forming intermediate filaments in the cell, as a possible target against HIV infection. We demonstrated a strong reduction of an HIV-1 based lentivirus expressing the enhanced green fluorescent protein (eGFP) in vimentin knockdown cells, and a noteworthy decrease of HIV-1 capsid protein antigen (CAp24) in those cells using a multiround infectivity assay. Electron micrographs showed changes in the structure of intermediate filaments when MT4 cells were treated with an anti-HIV leukocyte extract. Changes in the structure of intermediate filaments were also observed in vimentin knockdown MT4 cells. A synthetic peptide derived from a cytoskeleton protein showed potent inhibitory activity on HIV-1 infection, and low cytotoxicity. Our data suggest that vimentin can be a suitable target to inhibit HIV-1.


Clinico-Pathological Spectrum of Alveolar Soft Part Sarcoma: Case Series from a Tertiary Care Cancer Referral Centre in India with a Focus on Unusual Clinical and Histological Features.
Alveolar soft part sarcoma (ASPS) is characterized by distinctive histomorphology of variably discohesive epithelioid cells arranged in nests and translocation of t(x;17) (p11.2;q25) resulting in ASPSCR1-TFE3 fusion. The aim of the present study is to review the clinical, histopathological, and immunohistochemical profile of ASPS with a focus on unusual histological features. The present study is retrospective and descriptive. All cases with a diagnosis of ASPS were retrieved with clinical and radiology details. 22 patients of ASPS were identified. The most common site was the lower extremity and the size range was 3-22 cm. 54.5% of the patients had metastasis, with the lung as the most common site. Metastasis preceded detection of primary tumour in two cases. All cases showed similar histopathology of monomorphic epithelioid cells arranged in nests encircled by sinusoidal vasculature. Architecturally, the organoid pattern (81.8%) was followed by the alveolar pattern. 68.2% of the cases showed apple bite nuclei as the predominant nuclear feature. Rare nuclear features included binucleation (n=13), multinucleation (n=8), pleomorphism (n=4), nuclear grooves in three cases and intranuclear inclusion in one case, mitosis (n=5), and focal necrosis (n=6). All cases were positive for TFE3 and negative for AE1/AE3, EMA, HMB45, PAX8, MyoD1, SMA, synaptophysin, and chromogranin. Only two cases showed focal S100 positivity while one showed focal desmin positivity. Diffuse strong nuclear TFE3 positivity is sensitive for ASPS in an appropriate clinicoradiological context. Due to the high propensity for early metastasis, complete metastatic work-up and long term follow up is recommended.


The physiological and morphological phenotype of a yeast mutant resistant to the quaternary ammonium salt N-(dodecyloxycarboxymethyl)-N,N,N-trimethyl ammonium chloride.
We investigated the action of the quaternary ammonium salt (QAS) called IM (N-(dodecyloxycarboxymethyl)-N,N,N-trimethyl ammonium chloride) on Saccharomyces cerevisiae yeast cells. Changes in the yeast cell ultrastructure were confirmed by electron microscopy. We treated resistant mutant cells with QAS, and confirmed destruction of the mutant cytoplasm, an increase in the thickness of the cell wall, separation of the cell wall from the cytoplasm, and the accumulation of numerous lipid droplets. We also observed a relatively high production of lipids in the cells of the parental wild-type strain Sigma1278b and in its IM-resistant (IM(R)) mutant in the presence of the QAS. The IM(R) mutant showed increased sensitivity to CaCl(2) and SDS, and resistance to ethidium bromide, chloramphenicol, erythromycin and osmotic shock. It also tolerated growth at low pH. We suggest that the resistance to IM could be connected with the level of permeability of the cell membrane because the IM(R) mutant was sensitive to this compound in vivo in the presence of SDS and guanidine hydrochloride, which cause increased permeability of the cell plasma membrane.


Social inequalities in adult male mortality by the National Statistics Socio-Economic Classification, England and Wales, 2001-03.
This article reports social inequalities in mortality in men aged 25-64 years in England and Wales, in the period 2001-03 using unlinked data sources and 2001-04 using linked data sources. It represents the first official analysis of premature mortality by the final version of the National Statistics Socio-economic Classification introduced in 2001, and updates the tradition of decennial reporting of mortality by socio-economic status. These results set a benchmark for inequalities in mortality in men of this age, providing insights into the impacts of different social and occupational circumstances in the early 21st Century and enabling future monitoring. The Office for National Statistics intends to extend this work to examine inequalities to mortality in females of the same age, in causes of death and by Government Office Region.


Assessment of total retinal blood flow using Doppler Fourier Domain Optical Coherence Tomography during systemic hypercapnia and hypocapnia.
The purpose of this study was to investigate changes in total retinal blood flow (RBF) using Doppler Fourier Domain Optical Coherence Tomography (Doppler FD-OCT) in response to the manipulation of systemic partial pressure of CO2 (PETCO2). Double circular Doppler blood flow scans were captured in nine healthy individuals (mean age ± standard deviation: 27.1 ± 4.1, six males) using the RTVue(™) FD-OCT (Optovue). PETCO2 was manipulated using a custom-designed computer-controlled gas blender (RespirAct(™)) connected to a sequential gas delivery rebreathing circuit. Doppler FD-OCT measurements were captured at baseline, during stages of hypercapnia (+5/+10/+15 mmHg PETCO2), return to baseline and during stages of hypocapnia (-5/-10/-15 mmHg PETCO2). Repeated measures analysis of variance (reANOVA) and Tukey's post hoc analysis were used to compare Doppler FD-OCT measurements between the various PETCO2 levels relative to baseline. The effect of PETCO2 on TRBF was also investigated using linear regression models. The average RBF significantly increased by 15% (P < 0.0001) with an increase in PETCO2 and decreased significantly by 10% with a decrease in PETCO2 (P = 0.001). Venous velocity significantly increased by 3.11% from baseline to extreme hypercapnia (P < 0.001) and reduced significantly by 2.01% at extreme hypocapnia (P = 0.012). No significant changes were found in the average venous area measurements under hypercapnia (P = 0.36) or hypocapnia (P = 0.40). Overall, increased and decreased PETCO2 values had a significant effect on RBF outcomes (P < 0.002). In healthy individuals, altered end-tidal CO2 levels significantly changed RBF as measured by Doppler FD-OCT.


Retinal neuroprotection by hypoxic preconditioning is independent of hypoxia-inducible factor-1 alpha expression in photoreceptors.
Hypoxic preconditioning stabilizes hypoxia-inducible factor (HIF) 1 alpha in the retina and protects photoreceptors against light-induced cell death. HIF-1 alpha is one of the major transcription factors responding to low oxygen tension and can differentially regulate a large number of target genes. To analyse whether photoreceptor-specific expression of HIF-1 alpha is essential to protect photoreceptors by hypoxic preconditioning, we knocked down expression of HIF-1 alpha specifically in photoreceptor cells, using the cyclization recombinase (Cre)-lox system. The Cre-mediated knockdown caused a 20-fold reduced expression of Hif-1 alpha in the photoreceptor cell layer. In the total retina, RNA expression was reduced by 65%, and hypoxic preconditioning led to only a small increase in HIF-1 alpha protein levels. Accordingly, HIF-1 target gene expression after hypoxia was significantly diminished. Retinas of Hif-1 alpha knockdown animals did not show any pathological alterations, and tolerated hypoxic exposure in a comparable way to wild-type retinas. Importantly, the strong neuroprotective effect of hypoxic preconditioning against light-induced photoreceptor degeneration persisted in knockdown mice, suggesting that hypoxia-mediated survival of light exposure does not depend on an autocrine action of HIF-1 alpha in photoreceptor cells. Hypoxia-mediated stabilization of HIF-2 alpha and phosphorylation of signal transducer and activator of transcription 3 (STAT 3) were not affected in the retinas of Hif-1 alpha knockdown mice. Thus, these factors are candidates for regulating the resistance of photoreceptors to light damage after hypoxic preconditioning, along with several potentially neuroprotective genes that were similarly induced in hypoxic knockdown and control mice.


Prolongation of overall treatment time as a cause of treatment failure in early breast cancer: An analysis of the UK START (Standardisation of Breast Radiotherapy) trials of radiotherapy fractionation.
Tests of tumour treatment time effect in patients prescribed post-operative radiotherapy for early breast cancer have focussed on time to start of radiotherapy rather than overall treatment time. The START randomised trials of radiotherapy fractionation provide an opportunity to directly estimate the effect of treatment acceleration. Between 1986 and 2002, a total of 5861 women with early breast cancer were recruited into the UK START pilot (START-P), START-A and START-B randomised trials. START-P and START-A tested 13 fractions of 3.0-3.3Gy against 25 fractions of 2.0Gy with a fixed treatment duration of 5weeks for all schedules; START-B tested 15 fractions of 2.67Gy in 3weeks against 25 fractions of 2.0Gy over 5weeks. Estimates of the effect of length of treatment for local-regional relapse and for a measure of late normal tissue effects (change in photographic breast appearance, for patients following breast conserving surgery) were obtained from Cox proportional hazards regression analyses stratified according to trial. At a median follow-up of 10years, 444/5831 (7.6%) patients with data available had a local-regional relapse, and 1135/3185 (35.6%) had mild or marked change in photographic breast appearance by 5years. Adjusting for prognostic factors, the estimate of the overall treatment time effect for local-regional relapse was 0.60Gy/day (95%CI 0.10 to 1.18Gy/day, p=0.02), and 0.14Gy/day (95%CI -0.09 to 0.34Gy/day, p=0.29) for change in photographic breast appearance. Combined analysis of the START trials generates the hypothesis that overall treatment time is a significant determinant of local cancer control after adjuvant whole breast radiotherapy, with approximately 0.6Gy per day 'wasted' in compensating for tumour cell proliferation.


In vivo gene delivery and visualization of corneal stromal cells using an adenoviral vector and keratocyte-specific promoter.
This study was conducted to determine whether intrastromal injection of adenoviral construct could be used to transfect corneal stroma cells effectively in vivo and to determine whether a tissue-specific promoter could be used to express exogenous genes in keratocytes. An adenoviral construct with a cytomegalovirus (pCMV)-driven enhanced green fluorescent protein (EGFP) reporter gene was injected into the stroma of murine corneas. In vivo expression was quantitated and samples were analyzed by in vivo stereomicroscopy, and ex vivo expression was determined by confocal three dimensional (3-D) reconstruction. The 3.2-kb keratocan promoter was used to drive tissue-specific reporter gene expression in vivo. EGFP expression was first detected in vivo 11 hours after injection of adeno-EGFP in the corneal stroma, with a duration of approximately 3 weeks. Ex vivo wholemount cornea confocal analysis with 3-D reconstruction allowed visualization of EGFP expression in corneal stroma cells, to accurately assess cellular architecture and distribution in the corneal stroma. Naked pCMV-EGFP plasmid DNA did not express the reporter gene to the levels of the adeno-EGFP. The 3.2-kb keratocan promoter was capable of driving EGFP tissue-specific expression in the cornea. Intrastromal injection of adenovirus packaged DNA constructs is a rapid and efficient way to deliver and express genes in the corneal stroma. Intrastromal injection is also capable of delivering tissue-specific promoter constructs to the corneal stroma for gene expression. Furthermore, 3-D reconstruction provides a powerful tool for enhanced visualization of the corneal stroma environment and cellular biology.


Fabrication of ultrathin membrane via layer-by-layer self-assembly driven by hydrophobic interaction towards high separation performance.
A novel and facile layer-by-layer (LbL) self-assembly process driven by hydrophobic interaction and then reinforced by hydrogen bond was developed to prepare ultrathin membranes. Gelatin (GE) and tannic acid (TA) were alternately deposited on polyacrylonitrile (PAN) ultrafiltration membranes to obtain GE/TA membranes. The required number of deposition cycles for acceptable permselectivity of membrane was greatly reduced compared with that of the traditional LbL self-assembly process and could be ascribed to the rapid growth of membrane thickness and the integrity of the innermost gelatin layer. Higher surface hydrophilicity and more appropriate free volume characteristics were obtained for GE/TA multilayer membranes compared with pristine gelatin membrane. Moreover, the GE/TA multilayer membrane exhibited improved stability even at high water content of 30 wt %. The membrane separation experiments with pervaporation dehydration of ethanol aqueous solution as a model system demonstrated the GE/TA multilayer membrane achieved higher water permselectivity than the pristine gelatin membrane. High operation stability was acquired in the long-term membrane separation test.


Transparent, well-aligned TiO(2) nanotube arrays with controllable dimensions on glass substrates for photocatalytic applications.
Transparent, well-aligned TiO(2) nanotube arrays (NTAs) with controllable dimensions are grown on glass substrates via atomic layer deposition (ALD) of TiO(2) onto free-standing porous anodic alumina (PAA) templates. Photodegradation of aqueous methylene blue (MB) solution and solid stearic acid (SA) film using TiO(2) NTAs of various wall thicknesses are investigated. The Pd functionalized TiO(2) NTAs, with a wall thickness of 15 nm and height of 200 nm, has the highest photodegradation efficiency at 76% after 4 h of UV irradiation. These functionalized NTAs are able to photodegrade MB molecules completely as no obvious demethylated byproducts are observed during the process. It also demonstrates excellent photocatalytic activity for solid contaminants such as SA film. By using the ALD technique, the nanotube wall thickness can be precisely controlled so that it is sufficiently thin to be transparent while sufficiently thick for excellent photocatalytic performances. The transparent TiO(2) NTAs on glass substrates with excellent photocatalytic properties might have potential applications in self-cleaning coating, transparent electronics, and solar cells.


The validity of DSM-IV-TR criteria B and C of hair-pulling disorder (trichotillomania): evidence from a clinical study.
In both DSM-IV-TR and the ICD-10, hair-pulling disorder (trichotillomania, or TTM) is described as hair-pulling, with a rising urge or tension prior to pulling or when attempting to resist, and pleasure, relief or gratification during or after pulling. However, it has been questioned whether all patients with hair-pulling experience these other phenomena, and whether they occur with all pulling episodes. The objective of this study was to examine the DSM-IV-TR requirement of criteria B and C for a diagnosis of TTM in a sample of people with hair-pulling. A multi-site sample of adults with hair-pulling who met both DSM-IV-TR diagnostic criteria B and C (n=82, 89.13%) were compared to those who failed to satisfy both B and C (n=10, 10.87%) on a number of clinical variables. There were no differences in hair-pulling severity, levels of comorbid depressive and anxiety symptoms, number of comorbid body-focused repetitive behaviors, or impairment between those patients who did and did not meet criteria B and C. Our study does not provide convincing support for the inclusion of the current diagnostic criteria B and C for TTM in DSM-5.


Circulating fibrocytes are increased in children and young adults with pulmonary hypertension.
Chronic inflammation is an important component of the fibroproliferative changes that characterise pulmonary hypertensive vasculopathy. Fibrocytes contribute to tissue remodelling in settings of chronic inflammation, including animal models of pulmonary hypertension (PH). We sought to determine whether circulating fibrocytes were increased in children and young adults with PH. 26 individuals with PH and 10 with normal cardiac anatomy were studied. Fresh blood was analysed by flow cytometry for fibrocytes expressing CD45 and procollagen. Fibrocyte numbers were correlated to clinical and haemodynamic parameters, and circulating CC chemokine ligand (CCL)2 and CXC chemokine ligand (CXCL)12 levels. We found an enrichment of circulating fibrocytes among those with PH. No differences in fibrocytes were observed among those with idiopathic versus secondary PH. Higher fibrocytes correlated to increasing mean pulmonary artery pressure and age, but not to length or type of treatment. Immunofluorescence analysis confirmed flow sorting specificity. Differences in plasma levels of CCL2 or CXCL12, which could mobilise fibrocytes from the bone marrow, were not found. We conclude that circulating fibrocytes are significantly increased in individuals with PH compared with controls. We speculate that these cells might play important roles in vascular remodelling in children and young adults with pulmonary hypertension.


Calcareous nannoplankton response to surface-water acidification around Oceanic Anoxic Event 1a.
Ocean acidification induced by atmospheric CO2 may be a major threat to marine ecosystems, particularly to calcareous nannoplankton. We show that, during the Aptian (approximately 120 million years ago) Oceanic Anoxic Event 1a, which resulted from a massive addition of volcanic CO2, the morphological features of calcareous nannofossils traced the biological response to acidified surface waters. We observe the demise of heavily calcified nannoconids and reduced calcite paleofluxes at the beginning of a pre-anoxia calcification crisis. Ephemeral coccolith dwarfism and malformation represent species-specific adjustments to survive lower pH, whereas later, abundance peaks indicate intermittent alkalinity recovery. Deepwater acidification occurred with a delay of 25,000 to 30,000 years. After the dissolution climax, nannoplankton and carbonate recovery developed over approximately 160,000 years under persisting global dysoxia-anoxia.


Psychological factors in the genesis and management of nonepileptic seizures: clinical observations.
Nonepileptic seizures (NES) are frequently thought to have a "psychogenic" basis. Two 6-month group psychotherapy programs were provided for patients diagnosed as having NES [eight patients were treated during the first program, seven during the second (N=15)] to explore the potential role of psychological factors in the genesis of NES and to determine if psychotherapeutic interventions reduced the frequency of NES. Of the 15 patients, 9 (60%) completed at least 58% of the treatment sessions. Of those 9 patients, 6 (66%) reported a decline in "seizure frequency." One reported an increase (11%). Self-reported frequency highly correlated with paranoid ideation. Dissociative phenomena were common as was a history of sexual abuse. Each patient reported being in an adult situation that they found unacceptable or intolerable. None perceived a solution to their situation. Reports by health care providers that their seizures were not "real" (i.e., true epilepsy) restimulated feelings associated with their not being believed when they reported being sexually abused as children. The psychological genesis of NES in this sample of patients appears rooted in the recurrent experience of being in abusive or exploited relationships for which they perceived no solution.


Localization of the initial developmental stages of Loma salmonae in rainbow trout (Oncorhynchus mykiss).
The intracellular microsporidian parasite Loma salmonae affects salmonids of the genus Oncorhynchus and is a significant cause of economic losses in pen-reared Chinook salmon (O. tshawytscha) in British Columbia. Loma salmonae infection is easily recognized by the xenomas that form in the gills, but early stages of infection are difficult to detect in histologic sections. In situ hybridization (ISH), using an L. salmonae-specific digoxigenin-labeled single-stranded DNA probe, was used to detect the parasite during the early stages of infection. Loma salmonae was detected in the gut mucosal epithelium as early as 24 hours postexposure (PE), and it localized in the lamina propria of the intestine within 24 hours of infection. After the parasite was detected in the lamina propria, dividing merogonic stages in infected cells in the heart were detected by ISH as early as 2 days PE, providing the first evidence of parasitaemia and hematogenous distribution of this parasite in infected blood cells. The parasites inside the infected cells appeared to be undergoing merogony as they passed through the heart, indicating that proliferation may start at the site of infection, before the parasite arrives to the gills for their final developmental phase. This is the first time that L. salmonae passage through the intestinal wall and migration to the heart has been visualized; however, the identity of the cells harboring the parasite has yet to be determined.


Phenotypic variation of Pseudomonas brassicacearum as a plant root-colonization strategy.
Pseudomonas brassicacearum was isolated as a major root-colonizing population from Arabidopsis thaliana. The strain NFM421 of P. brassicacearum undergoes phenotypic variation during A. thaliana and Brassica napus root colonization in vitro as well as in soil, resulting in different colony appearance on agar surfaces. Bacteria forming translucent colonies (phase II cells) essentially were localized at the surface of young roots and root tips, whereas wild-type cells (phase I cells) were localized at the basal part of roots. The ability of phase II cells to spread and colonize new sites on root surface correlates with over-production of flagellin as evidenced by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of surface proteins and microsequencing. Moreover, phase II cells showed a higher ability to swim and to swarm on semisolid agar medium. Phase I and phase II cells of P. brassicacearum NFM421 were tagged genetically with green fluorescent protein and red fluorescent protein. Confocal scanning laser microscopy was used to localize phase II cells on secondary roots and root tips of A. thaliana, whereas phase I cells essentially were localized at the basal part of roots. These experiments were conducted in vitro and in soil. Phenotypic variation on plant roots is likely to be a colonization strategy that may explain the high colonization power of P. brassicacearum.


Cost, health impacts and cost effectiveness of iceless refrigeration in India's last-mile vaccine cold chain delivery.
Compared with ice-based vaccine carriers (IBVCs), iceless vaccine carrier (ILVC) last-mile delivery could optimize vaccine effectiveness by reducing spoilage. We estimated ILVC-associated spoilage costs averted and cost effectiveness. IBVC vaccine spoilage costs were estimated for six vaccines. ILVC incremental costs were based on yearly ILVC cost over total doses. Cost effectiveness was estimated via Markov modeling of rotavirus vaccine. The spoilage cost using IBVCs was US$9 603 294. Using ILVCs, the incremental cost per vaccine dose was US$0.026, the cost-benefit ratio was 0.28, the number of averted disability-adjusted life years was 0.03 per child and there was a saving of US$0.80 per child vaccinated. ILVCs may bring cost savings and health gains compared with IBVCs.


A support programme for caregivers of children with disabilities in Ghana: Understanding the impact on the wellbeing of caregivers.
Four fifths of the estimated 150 million children with disability in the world live in resource poor settings where the role of the family is crucial in ensuring that these children survive and thrive. Despite their critical role, evidence is lacking on how to provide optimal support to these families. This study explores the impact of a participatory training programme for caregivers delivered through a local support group, with a focus on understanding caregiver wellbeing. A qualitative longitudinal study was conducted to investigate the impact of a training programme, "getting to know cerebral palsy," with caregivers on their wellbeing. Eighteen caregivers, from four districts, were interviewed up to three times over 14 months, to assess impact and the reasons for any changes. Low levels of knowledge, high levels of stigma, physical and emotional exhaustion, and often difficult family relationships with social exclusion of the child and caregiver were common themes at the outset. Caregivers struggled to combine their caring and economic activities. This was exacerbated by the common absence of the father. Two months after completion of the training, their reported wellbeing had improved. The reasons for this were an improved understanding about their child's condition, positive attitudinal change towards their child, feelings of hope, and through the group support, a profound realisation that they are "not on their own." While relationships within the family remained complex in many cases, the support group offered an important and alternative social support network. This study illustrates the many benefits of a relatively simple caregiver intervention, which has the potential to offer a mechanism to provide sustainable social support for caregivers and children with cerebral palsy. Any future programme needs to also address more structural issues, including stigma and discrimination, and strengthen approaches to family engagement.


Chemical and chromatographic analysis of lipopolysaccharide from an antibiotic-supersusceptible mutant of Pseudomonas aeruginosa.
Lipopolysaccharides extracted from Pseudomonas aeruginosa strain K799 and its antibiotic-supersusceptible derivative Z61 were analyzed chemically and chromatographically. The side-chain polysaccharides purified by gel exclusion chromatography were compositionally identical, being composed of fucosamine (2-amino-2,6-dideoxygalactose), quinovosamine (2-amino-2,6-dideoxyglucose), and an unidentified amino sugar. In addition, low amounts of the core-specific components (glucose, rhamnose, alanine, and galactosamine) were found associated with the side chains from both strains. An average molecular weight of 38,000 to 50,000 was calculated for this fraction based on the glucose and rhamnose levels. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the lipopolysaccharides from these two strains were microheterogeneous. Qualitative analysis of the lipopolysaccharide neutral sugars, using a series of single-step revertants of mutant Z61, demonstrated that full revertants showed patterns indistinguishable from those of the wild-type strain K799, whereas partial revertants had intermediate levels and mutant Z61 low levels of neutral sugars. Quantitative analysis revealed that the core oligosaccharide fraction from the wild-type strain had a glucose/rhamnose/galactosamine ratio of 4:1:1, whereas the core from Z61 exhibited major deficiencies in both glucose and rhamnose. The lipid A from both strains contained five fatty acids, namely, 3-hydroxydecanoate, dodecanoate, 2- and 3-hydroxydodecanoate, and hexadecanoate. Whereas the overall fatty acid content was equal, the mutant strain showed markedly lower levels of dodecanoate and hexadecanoate and increased levels of 2-hydroxydodecanoate. Results of whole-cell fatty acid analyses were consistent with this observation. Evidence for an additional alteration of the lipid A of strain Z61 was obtained from acid hydrolysis studies and infrared spectra of isolated lipid A, although the actual chemical basis could not be determined by a variety of techniques. It is suggested that the state of the lipopolysaccharide is able to influence the number of open functional protein F pores in the outer membrane of P. aeruginosa.


Protection from La Crosse virus encephalitis with recombinant glycoproteins: role of neutralizing anti-G1 antibodies.
La Crosse virus, a member of the California serogroup of bunyaviruses, is an important cause of pediatric encephalitis in the midwestern United States. Like all bunyaviruses, La Crosse virus contains two glycoproteins, G1 and G2, the larger of which, G1, is the target of neutralizing antibodies. To develop an understanding of the role of each of the glycoproteins in the generation of a protective immune response, we immunized 1-week-old mice with three different preparations: a vaccinia virus recombinant (VV.ORF) that expresses both G1 and G2, a vaccinia virus recombinant (VV.G1) that expresses G1 only, and a truncated soluble G1 (sG1) protein prepared in a baculovirus system. Whereas VV.ORF generated a protective response that was mostly directed against G1, VV.G1 was only partially effective at inducing a neutralizing response and at protecting mice from a potentially lethal challenge with La Crosse virus. Nevertheless, a single immunization with the sG1 preparation resulted in a robust immune response and protection against La Crosse virus. These results indicate that (i) the G1 protein by itself can induce an immune response sufficient for protection from a lethal challenge with La Crosse virus, (ii) a neutralizing humoral response correlates with protection, and (iii) the context in which G1 is presented affects its immunogenicity. The key step in the defense against central nervous system infection appeared to be interruption of a transient viremia that occurred just after La Crosse virus inoculation.


Injury prevention in child death review: child pedestrian fatalities.
This article describes the epidemiology of child pedestrian fatalities in British Columbia using data generated by the province's Child Death Review Unit, to demonstrate the unique capacity of child death review to provide an ecological understanding of child mortality and catalyse evidence based, multi-level prevention strategies. All child pedestrian fatalities in British Columbia from 1 January 1 2003 to 31 December 2008 were reviewed. Data on demographics, circumstance of injury, and risk factors related to the child, driver, vehicle, and physical environment were extracted. Frequency of sociodemographic variables and modifiable risk factors were calculated, followed by statistical comparisons against the general population for Aboriginal ancestry, gender, ethnicity, income assistance and driver violations using z and t tests. Analysis of child pedestrian fatalities (n=33) found a significant overrepresentation of Aboriginal children (p=0.06), males (p<0.01), and children within low income families (p<0.01). The majority of incidents occurred in residential areas (51.5%), with a speed limit of 50 kph or higher (85.7%). Risky pedestrian behaviour was a factor in 56.7% of cases, and 33% of children under 10 years of age were not under active supervision. Drivers had significantly more driving violations than the comparison population (p<0.01). Child pedestrian fatalities are highly preventable through the modification of behavioural, social, and environmental risk factors. This paper illustrates the ability of child death review to generate an ecological understanding of injury epidemiology not otherwise available and advance policy and programme interventions designed to reduce preventable child mortality.


Predictors of patient-initiated reconsultation for lower respiratory tract infections in general practice.
Reconsultation for lower respiratory tract infection (LRTI) is common in general practice, but those who reconsult rarely have more significant illness warranting antibiotics. Knowledge of factors that predict patient-initiated reconsultation may allow clinicians to address specific issues during the initial consultation that could reduce reconsultations. Thirty-three per cent of a cohort of 431 LRTI patients in a randomised controlled trial reconsulted. Excluding 35 patients with GP-requested reconsultation left 28% (112/396) with a patient-initiated reconsultation during 28-day follow-up. Patient-reported dyspnoea and concerns that persisted after the initial consultation independently predicted patient-initiated reconsultation.


Morphological and Molecular Identification of Cytospora germanica Causing Canker on Populus spp. in China.
Species of Cytospora Ehrenb. cause canker and dieback on many genera of hardwoods. During surveys of forest trees in 2004 and 2008, some hardwoods such as Populus spp. with symptoms of canker and dieback were found in Liaoning and Xinjiang provinces, respectively. In these trees, the canker pathogen discolored the sapwood. During wet weather, the conidia were exuded from the fruiting bodies in gelatinous matrices, usually as yellow or orange tendrils. In the spring, the affected trees were wilting and the diffuse cankers spread rapidly and extensively during the period when trees began active spring growth. For saplings, sometimes the death rate of the pathogen has exceeded 50%. Conidiomatal stromata immersed in bark, prominent, circular to ovoid, 1.10 ± 0.23 mm in diameter (n = 10). Discs were white, nearly flat, circular, 0.44 ± 0.04 mm in diameter (n = 10), with one or two ostioles per disc. Ostioles were gray. Locules were subdivided by invaginations into chambers. Conidia were hyaline and lelongate-allantoid shaped, 4 to 5.4 μm long and 1 to 1.4 μm wide. Pieces (5 × 5 mm2) of the junction of affected and healthy tissues were surface sterilized with 1% NaOCl for 30 s and then rinsed twice in sterile distilled water. The fragments were placed on potato dextrose agar (PDA) plates incubated at 25°C for 7 days. The obtained isolates were cultured on PDA at 25°C in diffuse fluorescent light for 30 days, and then were deposited in the culture collection of the Chinese Academy of Forestry. According to these morphological features we initially thought that this pathogen was Cytospora chrysosperma (Pers.) Fr. However, BLAST analysis showed 98% and 99% homology with ITS sequence of isolate CBS118560 (GenBank Accession No. DQ243793), 99% and 100% homology with LSU gene of isolate AR3427 (AF362561) when we amplified ITS-rDNA gene and LSU gene of the isolates. The sequences were submitted to GenBank with the following accession numbers: JQ086563, JQ086564, JX524617, and JX524618. Pathogenicity tests were conducted in the greenhouse by inoculating 20 disinfected (70% ethanol) Populus tomentosa cuttings. Another two cuttings were treated with water agar as controls. The cuttings were incubated at 25°C for 30 days. In 18 of the 20 cuttings, the cambium became brown and appeared water soaked 20 days later, whereas controls did not show any symptoms. C. germanica was reisolated from symptomatic tissues. Inoculations were later repeated two times with similar results. Hubbes (1) and Spielman (2) considered V. germanica (teleomorph of C. germanica) a synonym of V. sordida (teleomorph of C. chrysosperma) based on morphological studies. We almost regarded these isolates as C. chrysosperma too. Morphological identification has been a weak and often inadequate means of identifying Cytospora species (1,2); the present study based on ITS-rDNA gene and LSU gene has substantially elevated identifications. To our knowledge, this is the first report of C. germanica on Populus spp. in China. Populus species are economically important trees in forests of North China and C. germanica has the potential to cause significant economic losses. References: (1) M. Hubbes. Phytopathol. Z. 39:65, 1960. (2) L. J. Spielman. Can. J. Bot. 63:1355, 1985.