Catheter-based renal denervation for resistant hypertension: 12-month results of the EnligHTN I first-in-human study using a multielectrode ablation system.
Renal denervation has emerged as a novel approach for the treatment of patients with drug-resistant hypertension. To date, only limited data have been published using multielectrode radiofrequency ablation systems. In this article, we present the 12-month data of EnligHTN I, a first-in-human study using a multielectrode ablation catheter. EnligHTN I enrolled 46 patients (average age, 60±10 years; on average 4.7±1.0 medications) with drug-resistant hypertension. Eligible patients were on ≥3 antihypertensive medications and had a systolic blood pressure (BP) ≥160 mm Hg (≥150 mm Hg for diabetics). Bilateral renal artery ablation was performed using a percutaneous femoral approach and standardized techniques. The average baseline office BP was 176/96 mm Hg, average 24-hour ambulatory BP was 150/83 mm Hg, and average home BP was 158/90 mm Hg. The average reductions (mm Hg) at 1, 3, 6, and 12 months were as follows: office: -28/-10, -27/-10, -26/-10, and -27/-11 mm Hg (P<0.001 for all); 24-hour ambulatory: -10/-5, -10/-5, -10/-6 (P<0.001 for all), and -7/-4 for 12 months (P<0.0094). Reductions in home measurements (based on 2-week average) were -9/-4, -8/-5,-10/-7, and -11/-6 mm Hg (P<0.001 at 12 months). At 12 months, there were no signals of worsening renal function and no new serious or life-threatening adverse events. One patient with baseline nonocclusive renal artery stenosis progressed to 75% diameter stenosis, requiring renal artery stenting. The 12-month data continue to demonstrate safety and efficacy of the EnligHTN ablation system in patients with drug-resistant hypertension. Home BP measurements parallel measurements obtained with 24-hour ambulatory monitoring.


Carbon monoxide from heme oxygenase-2 Is a tonic regulator against NO-dependent vasodilatation in the adult rat cerebral microcirculation.
Although the brain generates NO and carbon monoxide (CO), it is unknown how these gases and their enzyme systems interact with each other to regulate cerebrovascular function. We examined whether CO produced by heme oxygenase (HO) modulates generation and action of constitutive NO in the rat pial microcirculation. Immunohistochemical analyses indicated that HO-2 occurred in neurons and arachnoid trabecular cells, where NO synthase 1 (NOS1) was detectable, and also in vascular endothelium-expressing NOS3, suggesting colocalization of CO- and NO-generating sites. Intravital microscopy using a closed cranial window preparation revealed that blockade of the HO activity by zinc protoporphyrin IX significantly dilates arterioles. This vasodilatation depended on local NOS activities and was abolished by CO supplementation, suggesting that the gas derived from HO-2 tonically regulates NO-mediated vasodilatory response. Bioimaging of NO by laser-confocal microfluorography of diaminofluorescein indicated detectable amounts of NO at the microvascular wall, the subdural mesothelial cells, and arachnoid trabecular cells, which express NOS in and around the pial microvasculature. On CO inhibition by the HO inhibitor, regional NO formation was augmented in these cells. Such a pattern of accelerated NO formation depended on NOS activities and was again attenuated by the local CO supplementation. Studies using cultured porcine aortic endothelial cells suggested that the inhibitory action of CO on NOS could result from the photo-reversible gas binding to the prosthetic heme. Collectively, CO derived from HO-2 appears to serve as a tonic vasoregulator antagonizing NO-mediated vasodilatation in the rat cerebral microcirculation.


The relative efficacy of two interventions in altering maltreated preschool children's representational models: implications for attachment theory.
A narrative story-stem task was used to evaluate the efficacy of two competing, developmentally informed preventive interventions for maltreated preschoolers and their mothers designed to modify children's internal representations of self and of self in relation to other. One hundred and twenty-two mothers and their preschoolers (87 maltreated and 35 nonmaltreated) served as participants. Maltreating families were randomly assigned to either the preschooler-parent psychotherapy (PPP, n = 23), psychoeducational home visitation (PHV, n = 34), or community standard (CS, n = 30) intervention group at baseline. Thirty-five nonmaltreating (NC) families served as comparisons. Narratives were administered to children at baseline and at the postintervention evaluation. Children in the PPP intervention evidenced more of a decline in maladaptive maternal representations over time than PHV and CS children and displayed a greater decrease in negative self-representations than CS, PHV, and NC children. Also, the mother-child relationship expectations of PPP children became more positive over the course of the intervention, as compared to NC and PHV participants. These results suggest that an attachment-theory informed model of intervention (PPP) is more effective at improving representations of self and of caregivers than is a didactic model of intervention directed at parenting skills. Findings are discussed with respect to their implications for developmental theory, with a specific focus on attachment theory and internal working models of relationships.


Synthesis of oligonucleotides containing the N2-deoxyguanosine adduct of the dietary carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline.
2-amino-3-methylimidazo[4,5-f]quinoline (IQ) is a highly mutagenic heterocyclic amine formed in all cooked meats. IQ has been found to be a potent inducer of frameshift mutations in bacteria and carcinogenic in laboratory animals. Upon metabolic activation, IQ forms covalent adducts at the C8- and N2-positions of deoxyguanosine with a relative ratio of up to approximately 4:1. We have previously incorporated the major dGuo-C8-IQ adduct into oligonucleotides through the corresponding phosphoramidite reagent. We report here the sequence-specific synthesis of oligonucleotides containing the minor dGuo-N2-IQ adduct. Thermal melting analysis revealed that the dGuo-N2-IQ adduct significantly destabilizes duplex DNA.


Relaxation along a fictitious field (RAFF) and Z-spectroscopy using alternating-phase irradiation (ZAPI) in permanent focal cerebral ischemia in rat.
Cerebral ischemia alters the molecular dynamics and content of water in brain tissue, which is reflected in NMR relaxation, diffusion and magnetization transfer (MT) parameters. In this study, the behavior of two new MRI contrasts, Relaxation Along a Fictitious Field (RAFF) and Z-spectroscopy using Alternating-Phase Irradiation (ZAPI), were quantified together with conventional relaxation parameters (T1, T2 and T1ρ) and MT ratios in acute cerebral ischemia in rat. The right middle cerebral artery was permanently occluded and quantitative MRI data was acquired sequentially for the above parameters for up to 6 hours. The following conclusions were drawn: 1) Time-dependent changes in RAFF and T1ρ relaxation are not coupled to those in MT. 2) RAFF relaxation evolves more like transverse, rather than longitudinal relaxation. 3) MT measured with ZAPI is less sensitive to ischemia than conventional MT. 4) ZAPI data suggest alterations in the T2 distribution of macromolecules in acute cerebral ischemia. It was shown that both RAFF and ZAPI provide complementary MRI information from acute ischemic brain tissue. The presented multiparametric MRI data may aid in the assessment of brain tissue status early in ischemic stroke.


Investigation of multidrug-resistant fatal colisepticaemia in weanling pigs.
Escherichia coli is usually a benign commensal of the gut microflora. However, when E. coli acquires virulence genes it can multiply rapidly and cause disease through colonisation of the intestinal mucosa. Escherichia coli can become a significant pathogen in young pigs. We report an investigation of fatal colisepticaemia in weanling pigs from emerging farms where piglets and weaners were diarrhoeic and the mortality rate ranged between 15% and 70% in each litter. Faecal and tissue samples were processed for histopathology, bacteriology and molecular biology (multiplex and monoplex polymerase chain reaction) and we recovered enteroaggregative multidrug-resistant E. coli producing EAST-1 enterotoxin. An association between poor housing conditions and the observed cases was established and future management programmes were recommended to reduce the impact of such pathogens. Enteroaggregative E. coli is becoming a major problem in the pig industry. It therefore becomes necessary to establish the full impact of E. coli on the South African pig industry and to determine the geographic extent of the problem.


Contrasting roles of interallelic recombination at the HLA-A and HLA-B loci.
A statistical study of DNA sequences of alleles at the highly polymorphic class I MHC loci of humans, HLA-A and HLA-B, showed evidence of both large-scale recombination events (involving recombination of exons 1-2 of one allele with exons 3-8 of another) and small-scale recombination events (involving apparent exchange of short DNA segments). The latter events occurred disproportionately in the region of the gene encoding the antigen recognition site (ARS) of the class I molecule. Furthermore, they involved the ARS codons which are under the strongest selection favoring allelic diversity at the amino acid level. Thus, the frequency of recombinant alleles appears to have been increased by some form of balancing selection (such as overdominant selection) favoring heterozygosity in the ARS. These analyses also revealed a striking difference between the A and B loci. Recombination events appear to have occurred about twice as frequently at the B locus, and recombinants at the B locus were significantly more likely to affect polymorphic sites in the ARS. At the A locus, there are well-defined allelic lineages that have persisted since prior to the human-chimpanzee divergence; but at the B locus, there is no evidence for such long-lasting allelic lineages. Thus, relatively frequent interallelic recombination has apparently been a feature of the long-term evolution of the B locus but not of the A locus.


Sleep disturbances in preschool age children with cerebral palsy: a questionnaire study.
The study aimed to analyze (i) the prevalence of sleep disorders in pre-school children with cerebral palsy (CP) using the Sleep Disturbance Scale for Children (SDSC), (ii) the possible association with motor, cognitive and behavioral problems, and (iii) the possible differences with typically developing children matched for age and gender. One-hundred children with CP (age range: 3-5 years, mean: 3.8 years) were assessed using the SDSC, the Gross Motor Function Classification System (GMFCS), the Wechsler Preschool and Primary Scale of Intelligence, and the Child Behaviour Check List (CBCL) to assess sleep, motor, cognitive, and behavioral problems, respectively. Further 100 healthy children matched for age and sex were assessed using the SDSC. An abnormal total sleep score was found in 13% of children with CP while 35% had an abnormal score on at least one SDSC factor. SDSC total score was significantly associated with pathological internalizing scores on CBCL and active epilepsy on multivariate analysis. CP group reported higher significant median scores on SDSC total, parasomnias, and difficulty in initiating and maintaining sleep factors. In pre-school children sleep disorders are more common in children with CP than in healthy control group and are often associated with epilepsy and behavioral problems.


Tree species effects on ecosystem water-use efficiency in a high-elevation, subalpine forest.
Ecosystem water-use efficiency (eWUE; the ratio of net ecosystem productivity to evapotranspiration rate) is a complex landscape-scale parameter controlled by both physical and biological processes occurring in soil and plants. Leaf WUE (lWUE; the ratio of leaf CO(2) assimilation rate to transpiration rate) is controlled at short time scales principally by leaf stomatal dynamics and this control varies among plant species. Little is known about how leaf-scale variation in lWUE influences landscape-scale variation in eWUE. We analyzed approximately seven thousand 30-min averaged eddy covariance observations distributed across 9 years in order to assess eWUE in two neighboring forest communities. Mean eWUE was 19% lower for the community in which Engelmann spruce and subalpine fir were dominant, compared to the community in which lodgepole pine was dominant. Of that 19% difference, 8% was attributed to residual bias in the analysis that favored periods with slightly drier winds for the spruce-fir community. In an effort to explain the remaining 11% difference, we assessed patterns in lWUE using C isotope ratios. When we focused on bulk tissue from older needles we detected significant differences in lWUE among tree species and between upper and lower canopy needles. However, when these differences were scaled to reflect vertical and horizontal leaf area distributions within the two communities, they provided no power to explain differences in eWUE that we observed in the eddy covariance data. When we focused only on bulk needle tissue of current-year needles for 3 of the 9 years, we also observed differences in lWUE among species and in needles from upper and lower parts of the canopy. When these differences in lWUE were scaled to reflect leaf area distributions within the two communities, we were able to explain 6.3% of the differences in eWUE in 1 year (2006), but there was no power to explain differences in the other 2 years (2003 and 2007). When we examined sugars extracted from needles at 3 different times during the growing season of 2007, we could explain 3.8-6.0% of the differences in eWUE between the two communities, but the difference in eWUE obtained from the eddy covariance record, and averaged over the growing season for this single year, was 32%. Thus, overall, after accounting for species effects on lWUE, we could explain little of the difference in eWUE between the two forest communities observed in the eddy covariance record. It is likely that water and C fluxes from soil, understory plants, and non-needle tissues, account for most of the differences observed in the eddy covariance data. For those cases where we could explain some of the difference in eWUE on the basis of species effects, we partitioned the scaled patterns in lWUE into two components: a component that is independent of canopy leaf area distribution, and therefore only dependent on species-specific differences in needle physiology; and a component that is independent of species differences in needle physiology, and only dependent on species-specific influences on canopy leaf area distribution. Only the component that is dependent on species influences on canopy leaf area distribution, and independent of inherent species differences in needle physiology, had potential to explain differences in eWUE between the two communities. Thus, when tree species effects are important, canopy structure, rather than species-specific needle physiology, has more potential to explain patterns in eWUE.


Chromatin signature of embryonic pluripotency is established during genome activation.
After fertilization the embryonic genome is inactive until transcription is initiated during the maternal-zygotic transition. This transition coincides with the formation of pluripotent cells, which in mammals can be used to generate embryonic stem cells. To study the changes in chromatin structure that accompany pluripotency and genome activation, we mapped the genomic locations of histone H3 molecules bearing lysine trimethylation modifications before and after the maternal-zygotic transition in zebrafish. Histone H3 lysine 27 trimethylation (H3K27me3), which is repressive, and H3K4me3, which is activating, were not detected before the transition. After genome activation, more than 80% of genes were marked by H3K4me3, including many inactive developmental regulatory genes that were also marked by H3K27me3. Sequential chromatin immunoprecipitation demonstrated that the same promoter regions had both trimethylation marks. Such bivalent chromatin domains also exist in embryonic stem cells and are thought to poise genes for activation while keeping them repressed. Furthermore, we found many inactive genes that were uniquely marked by H3K4me3. Despite this activating modification, these monovalent genes were neither expressed nor stably bound by RNA polymerase II. Inspection of published data sets revealed similar monovalent domains in embryonic stem cells. Moreover, H3K4me3 marks could form in the absence of both sequence-specific transcriptional activators and stable association of RNA polymerase II, as indicated by the analysis of an inducible transgene. These results indicate that bivalent and monovalent domains might poise embryonic genes for activation and that the chromatin profile associated with pluripotency is established during the maternal-zygotic transition.


The inner nuclear membrane protein emerin regulates beta-catenin activity by restricting its accumulation in the nucleus.
Emerin is a type II inner nuclear membrane (INM) protein of unknown function. Emerin function is likely to be important because, when it is mutated, emerin promotes both skeletal muscle and heart defects. Here we show that one function of Emerin is to regulate the flux of beta-catenin, an important transcription coactivator, into the nucleus. Emerin interacts with beta-catenin through a conserved adenomatous polyposis coli (APC)-like domain. When GFP-emerin is expressed in HEK293 cells, beta-catenin is restricted to the cytoplasm and beta-catenin activity is inhibited. In contrast, expression of an emerin mutant, lacking its APC-like domain (GFP-emerinDelta), dominantly stimulates beta-catenin activity and increases nuclear accumulation of beta-catenin. Human fibroblasts that are null for emerin have an autostimulatory growth phenotype. This unusual growth phenotype arises through enhanced nuclear accumulation and activity of beta-catenin and can be replicated in wild-type fibroblasts by transfection with constitutively active beta-catenin. Our results support recent findings that suggest that INM proteins can influence signalling pathways by restricting access of transcription coactivators to the nucleus.


Sensitivity enhanced heteronuclear correlation spectroscopy in multidimensional solid-state NMR of oriented systems via chemical shift coherences.
We present new sensitivity enhanced schemes for heteronuclear correlation spectroscopy (HETCOR) in solid-state NMR of oriented systems. These schemes will enhance the sensitivity of the HETCOR by 40% for the two-dimensional experiments (SE-HETCOR) and up to 180% for the 3D HETCOR-separated local field version (SE-PISEMAI-HETCOR). The signal enhancement is demonstrated for a single crystal of ((15)N)N-acetylleucine and the integral membrane protein sarcolipin oriented in lipid bicelles. These methods will significantly reduce the time needed to acquire multidimensional experiments for membrane proteins oriented in magnetically or mechanically aligned lipid bilayers as well as liquid crystalline materials.


Blueberry polyphenols attenuate kainic acid-induced decrements in cognition and alter inflammatory gene expression in rat hippocampus.
Cognitive impairment in age-related neurodegenerative diseases such as Alzheimer's disease may be partly due to long-term exposure and increased susceptibility to inflammatory insults. In the current study, we investigated whether polyphenols in blueberries can reduce the deleterious effects of inflammation induced by central administration of kainic acid by altering the expression of genes associated with inflammation. To this end, 4-month-old male Fischer-344 (F344) rats were fed a control, 0.015% piroxicam (an NSAID) or 2% blueberry diet for 8 weeks before either Ringer's buffer or kainic acid was bilaterally micro-infused into the hippocampus. Two weeks later, following behavioral evaluation, the rats were killed and total RNA from the hippocampus was extracted and used in real-time quantitative RT-PCR (qRT-PCR) to analyze the expression of inflammation-related genes. Kainic acid had deleterious effects on cognitive behavior as kainic acid-injected rats on the control diet exhibited increased latencies to find a hidden platform in the Morris water maze compared to Ringer's buffer-injected rats and utilized non-spatial strategies during probe trials. The blueberry diet, and to a lesser degree the piroxicam diet, was able to improve cognitive performance. Immunohistochemical analyses of OX-6 expression revealed that kainic acid produced an inflammatory response by increasing the OX-6 positive areas in the hippocampus of kainic acid-injected rats. Kainic acid up-regulated the expression of the inflammatory cytokines IL-1beta and TNF-alpha, the neurotrophic factor IGF-1, and the transcription factor NF-kappaB. Blueberry and piroxicam supplementations were found to attenuate the kainic acid-induced increase in the expression of IL-1beta, TNF-alpha, and NF-kappaB, while only blueberry was able to augment the increased IGF-1 expression. These results indicate that blueberry polyphenols attenuate learning impairments following neurotoxic insult and exert anti-inflammatory actions, perhaps via alteration of gene expression.


Differential mRNA expression profiles in proximal tibia of aged rats in response to ovariectomy and low-Ca diet.
Both ovariectomized animals and animals fed with Ca-depleted diets are commonly used in vivo models for the investigation of osteoporosis-related bone loss. The present study aimed to study the genomic responses of bone in aged female rats to ovariectomy and dietary Ca deficiency in these models. Aged (11 months old) Sprague-Dawley rats were subjected to bilateral ovariectomy or sham-operation and fed with diets containing different dietary Ca content (LCD, 0.1% Ca or HCD, 1.2% Ca) for 12 weeks. Serum and urine were collected for biochemical marker measurement, and tibias were collected for bone mineral density (BMD) analysis by pQCT as well as for gene expression analysis by real-time PCR. Ovariectomy increased serum N-telopeptides of bone type I collagen (NTx) levels in aged rats fed with HCD (P<0.05). In addition, ovariectomy reduced BMD and predicted bone strength of tibial proximal metaphysis in aged rats fed with either LCD or HCD. Dietary Ca deficiency did not alter serum bone-specific alkaline phosphatase (BAP) or NTx levels, but induced a loss of BMD at tibia proximal metaphysis in aged rats. Ovariectomy promoted the mRNA expression of alpha-1 type I collagen (COL), osteoprotegerin (OPG) and receptor activator of nuclear factor-kappa B ligand (RANKL); and inhibited the mRNA expression of cathepsin K and matrix metalloproteinase-9 (MMP-9) in the proximal tibia of aged rats. Low-Ca diet significantly up-regulated the mRNA expression of COL, core binding factor I (Cbfa1), OPG and carbonic anhydrase II (CAII) in proximal tibia of aged rats. Our study revealed that the genomic responses of bone in proximal tibia to ovariectomy and dietary Ca deficiency were different. The bone loss induced by ovariectomy appears to be mediated primarily by an increase in RANKL mRNA expression; whereas the induction by dietary Ca restriction might be mediated by the induction of carbonic anhydrase II expression.


Reconnecting with oneself while struggling between life and death: The phenomenon of recovery as experienced by persons at risk of suicide.
The body of knowledge regarding health and recovery as experienced by patients at risk of suicide is limited. More research is needed into the meaning of recovery and what strengthens the desire to live. The aim of this study was to describe the phenomenon of recovery in a context of nursing care as experienced by persons at risk of suicide. In line with a reflective lifeworld research approach, 14 patients from a psychiatric clinic in Sweden participated in phenomenon-oriented interviews. Data were analyzed to describe the essence of the phenomenon. The results reveal that the phenomenon of recovery means 'reconnecting with oneself while struggling between life and death'. Three meaning constituents emerged: being in an expressive space and giving voice to oneself, regaining dignity through nurturing connectedness, and finding a balance in the tension between life and death. In conclusion, the meaning of recovery is to experience the ability to manage one's own life. Professional caregivers need to acknowledge patients' lifeworlds, in a way that enable patients to experience themselves as capable of managing their own lives. Professional caregivers should also facilitate the involvement of supportive relatives.


Fast and low-cost structured light pattern sequence projection.
We present a high-speed and low-cost approach for structured light pattern sequence projection. Using a fast rotating binary spatial light modulator, our method is potentially capable of projection frequencies in the kHz domain, while enabling pattern rasterization as low as 2 μm pixel size and inherently linear grayscale reproduction quantized at 12 bits/pixel or better. Due to the circular arrangement of the projected fringe patterns, we extend the widely used ray-plane triangulation method to ray-cone triangulation and provide a detailed description of the optical calibration procedure. Using the proposed projection concept in conjunction with the recently published coded phase shift (CPS) pattern sequence, we demonstrate high accuracy 3-D measurement at 200 Hz projection frequency and 20 Hz 3-D reconstruction rate.


Matrin 3 in neuromuscular disease: physiology and pathophysiology.
RNA-binding proteins (RBPs) are essential factors required for the physiological function of neurons, muscle, and other tissue types. In keeping with this, a growing body of genetic, clinical, and pathological evidence indicates that RBP dysfunction and/or gene mutation leads to neurodegeneration and myopathy. Here, we summarize the current understanding of matrin 3 (MATR3), a poorly understood RBP implicated not only in ALS and frontotemporal dementia but also in distal myopathy. We begin by reviewing MATR3's functions, its regulation, and how it may be involved in both sporadic and familial neuromuscular disease. We also discuss insights gleaned from cellular and animal models of MATR3 pathogenesis, the links between MATR3 and other disease-associated RBPs, and the mechanisms underlying RBP-mediated disorders.


Malaria knowledge and long-lasting insecticidal net use in rural communities of central Côte d'Ivoire.
To improve effectiveness of malaria control interventions, it is essential to deepen the knowledge of contextual factors that govern people's practice for preventive and curative measures. The aim of this study was to determine factors that influence the use of long-lasting insecticidal nets (LLINs) in three rural communities of Côte d'Ivoire, two of which benefited from recent interventions. The study was carried out in 957 households in three villages (Bozi, N'Dakonankro and Yoho) located in central Côte d'Ivoire. Indicators of socioeconomic position (SEP), malaria knowledge and practice, placing special emphasis on LLINs, were investigated during a cross-sectional questionnaire survey. Principal component analysis was used to calculate the SEP of households by means of a list of household assets ownership. The concentration index was used to assess the direction of the association between SEP and a given variable. To compare groups or means, Fisher's exact test, χ2 and Kruskal-Wallis test were used, as appropriate. Significant differences were found between SEP and reported malaria symptoms, such as fever or hot body, convulsion, anaemia and jaundice (yellow eyes). Individuals from the least poor group cited more often the use of bed nets and insecticide-treated nets (ITNs) compared to poorer groups. The mean number of individuals reporting the use of bed nets and LLINs was different between groups with different educational level. Moreover, the mean number of LLINs in a household was influenced by the presence of children below five years of age. The study not only confirmed that education and SEP play important roles in the prevention and control of malaria and promotion of health in general, but pointed at the basic essential knowledge and the key behavioural elements that should guide education and learning processes among the poorer segments of the population. In turn, such knowledge may change behaviour and lead to an increased utilization of LLINs.


An endoscopic system adopting a liquid crystal lens with an electrically tunable depth-of-field.
Conventional endoscopic systems consisting of several solid lenses suffer from a fixed and limited depth-of-field (DOF). For practical applications, conventional endoscopes mechanically change the distance between the solid lenses of a lens module in order to change the focusing plane and DOF to see clearly in a scene. In this paper, we demonstrate an electrically tunable endoscopic system adopting a liquid crystal lens. By means of tunable focusing properties of the LC lens as a positive lens and a negative lens, the object at different objective distances can be imaged to the image sensor clearly and the corresponding depth-of-field can also help to enlarge the total spatial depth perception in a scene. The optical mechanism is discussed. In the experiments, under adjustment of three discrete lens powers of the LC lens, the viewing range or total spatial depth perception of the endoscopic system is from 76.4 mm to 12.4 mm which is 2x improved compared to the conventional one without LC lens. We believe this study can be extended to the applications of industrial and medical endoscopes.


Back extensor muscle fatigue at submaximal workloads assessed using frequency banding of the electromyographic signal.
Changes in the mean or median frequency of the electromyographic (EMG) power spectrum are often used to assess skeletal muscle fatigue. A more global analysis of the spectral changes using frequency banding may provide a more sensitive measure of fatigue than changes in mean or median frequency. So, the aim of the present study was to characterize changes in different power spectrum frequency bands and compare these with changes in median frequency. Twenty male subjects performed isometric contractions of the back muscles in an isometric dynamometer at 30%, 40%, 50% and 60% of maximum voluntary contraction. During each contraction, surface EMG signals were recorded from the right and left longissimus thoracis muscles, and endurance time was measured. The EMG power spectra were divided into four frequency bands (20-50 Hz; 50-80 Hz; 80-110 Hz; 110-140 Hz) and changes in power in each band with fatigue were compared with changes in median frequency. The percentage changes in 20-50 Hz band were greater than in all other and the rate of change in power, indicated by the slope, was also greatest in 20-50 Hz band. Also, 20-50 Hz band had a greater change in power than the median frequency. Power in the low frequency part of the EMG power spectrum increases with fatigue in a load-dependent manner. The rate of change in low frequency power may be a useful indicator of fatigue rate or "fatigability" in the back muscles. Also, changes in low frequency power are more evident than changes in the median frequency.


Integration of sequence data from a Consanguineous family with genetic data from an outbred population identifies PLB1 as a candidate rheumatoid arthritis risk gene.
Integrating genetic data from families with highly penetrant forms of disease together with genetic data from outbred populations represents a promising strategy to uncover the complete frequency spectrum of risk alleles for complex traits such as rheumatoid arthritis (RA). Here, we demonstrate that rare, low-frequency and common alleles at one gene locus, phospholipase B1 (PLB1), might contribute to risk of RA in a 4-generation consanguineous pedigree (Middle Eastern ancestry) and also in unrelated individuals from the general population (European ancestry). Through identity-by-descent (IBD) mapping and whole-exome sequencing, we identified a non-synonymous c.2263G>C (p.G755R) mutation at the PLB1 gene on 2q23, which significantly co-segregated with RA in family members with a dominant mode of inheritance (P = 0.009). We further evaluated PLB1 variants and risk of RA using a GWAS meta-analysis of 8,875 RA cases and 29,367 controls of European ancestry. We identified significant contributions of two independent non-coding variants near PLB1 with risk of RA (rs116018341 [MAF = 0.042] and rs116541814 [MAF = 0.021], combined P = 3.2 × 10(-6)). Finally, we performed deep exon sequencing of PLB1 in 1,088 RA cases and 1,088 controls (European ancestry), and identified suggestive dispersion of rare protein-coding variant frequencies between cases and controls (P = 0.049 for C-alpha test and P = 0.055 for SKAT). Together, these data suggest that PLB1 is a candidate risk gene for RA. Future studies to characterize the full spectrum of genetic risk in the PLB1 genetic locus are warranted.


Chemical and immunological testing for faecal occult blood in screening subjects at risk of familial colorectal cancer.
People with a family history of colorectal cancer have an increased risk of the disease themselves. Many centres are advocating family history screening by endoscopy. The performance of chemical and immunological faecal occult blood tests (Haemoccult and Hemeselect) in 212 subjects with a family history of colorectal cancer was assessed. Both Hemeselect and Haemoccult were positive in the only patient with colorectal cancer. Hemeselect was more sensitive than Haemoccult for adenomas (40% compared with 20%) (adenomas larger than 1 cm 75% compared with 50%). No additional abnormality was detected by the addition of Haemoccult or Hemeselect to 60 cm flexible sigmoidoscopy in screening people at lower levels of familial risk. A false positive rate of 16% for Hemeselect resulted in a high proportion of additional colonoscopies in this group. At present faecal occult blood tests are not sufficiently sensitive or specific to replace endoscopy in screening people at risk of familial colorectal cancer.


Investigation of the effects of storage with preservatives at room temperature or refrigeration without preservatives on urinalysis results for samples from healthy dogs.
To compare urinalysis results for canine urine samples stored in preservative-containing tubes at room temperature (20°C to 25°C [68°F to 77°F]) or refrigerated at 4°C (39.2°F) in plain glass tubes with results for the same samples immediately after collection. Urine samples from 20 healthy dogs. Urine samples (1/dog) were divided into 6 aliquots (3 in preservative-containing tubes and 3 in plain glass tubes). Preservative-containing tubes were stored at room temperature and plain glass tubes were refrigerated. Urinalysis was performed 0, 24, and 72 hours after collection. Results for both storage conditions were compared with results for a reference sample (the 0-hour [immediate post-collection] aliquot in a plain glass tube) by Spearman correlation analysis with pairwise tests for selected variables. Physical variables (urine color and turbidity with and without centrifugation) for both storage conditions had high (rs = 0.7 to 0.9) or very high (rs = 0.9 to 1.0) degrees of positive correlation with reference sample results at all time points, except for color at 24 hours. Similar results were found for all biochemical variables with storage up to 72 hours. For microscopic characteristics, correlation with reference sample results ranged from low or nonsignificant to very high under both storage conditions. Results suggested that if a delay in urinalysis is expected, use of the preservative-containing tubes evaluated in this study may be a viable option for sample storage. Further research is warranted to assess direct comparability of results to those of freshly collected samples and use of these tubes to store samples from dogs with conditions affecting the urinary tract.


Interaction of R17 coat protein with its RNA binding site for translational repression.
The interaction between bacteriophage R17 coat protein and its RNA binding site for translational repression was studied as an example of a sequence-specific RNA-protein interaction. A nitrocellulose filter retention assay is used to demonstrate equimolar binding between the coat protein and a synthetic 21 nucleotide RNA fragment. The Kd at 2 degrees C in a buffer containing 0.19 M salt is about 1 nM. The relatively weak ionic strength dependence of Ka and a delta H = -19 kcal/mole indicates that most of the binding free energy is due to non-electrostatic interactions. Since a variety of RNAs failed to compete with the 21 nucleotide fragment for coat protein binding, the interaction appears highly sequence specific. We have synthesized more than 30 different variants of the binding site sequence in order to identify the portions of the RNA molecule which are important for protein binding. Out of the five single stranded residues examined, four were essential for protein binding whereas the fifth could be replaced by any nucleotide. One variant was found to bind better than the wild type sequence. Substitution of nucleotides which disrupted the secondary structure of the binding fragment resulted in very poor binding to the protein. These data indicated that there are several points of contact between the RNA and the protein and the correct hairpin secondary structure of the RNA is essential for protein binding.


Design of the xylitol for adult caries trial (X-ACT).
Dental caries incidence in adults is similar to that in children and adolescents, but few caries preventive agents have been evaluated for effectiveness in adults populations. In addition, dentists direct fewer preventive services to their adult patients. Xylitol, an over-the-counter sweetener, has shown some potential as a caries preventive agent, but the evidence for its effectiveness is not yet conclusive and is based largely on studies in child populations. X-ACT is a three-year, multi-center, placebo controlled, double-blind, randomized clinical trial that tests the effects of daily use of xylitol lozenges versus placebo lozenges on the prevention of adult caries. The trial has randomized 691 participants (ages 21-80) to the two arms. The primary outcome is the increment of cavitated lesions. This trial should help resolve the overall issue of the effectiveness of xylitol in preventing caries by contributing evidence with a low risk of bias. Just as importantly, the trial will provide much-needed information about the effectiveness of a promising caries prevention agent in adults. An effective xylitol-based caries prevention intervention would represent an easily disseminated method to extend caries prevention to individuals not receiving caries preventive treatment in the dental office. ClinicalTrials.Gov NCT00393055.


An organism-specific method to rank predicted coding regions in Trypanosoma brucei.
Genome annotation in differently evolved organisms presents challenges because the lack of sequence-based homology limits the ability to determine the function of putative coding regions. To provide an alternative to annotation by sequence homology, we developed a method that takes advantage of unusual trypanosomatid biology and skews in nucleotide composition between coding regions and upstream regions to rank putative open reading frames based on the likelihood of coding. The method is 93% accurate when tested on known genes. We have applied our method to the full complement of open reading frames on Chromosome I of Trypanosoma brucei, and we can predict with high confidence that 226 putative coding regions are likely to be functional. Methods such as the one described here for discriminating true coding regions are critical for genome annotation when other sources of evidence for function are limited.


A polybasic motif in ErbB3-binding protein 1 (EBP1) has key functions in nucleolar localization and polyphosphoinositide interaction.
Polyphosphoinositides (PPIns) are present in the nucleus where they participate in crucial nuclear processes, such as chromatin remodelling, transcription and mRNA processing. In a previous interactomics study, aimed to gain further insight into nuclear PPIns functions, we identified ErbB3 binding protein 1 (EBP1) as a potential nuclear PPIn-binding protein in a lipid pull-down screen. EBP1 is a ubiquitous and conserved protein, located in both the cytoplasm and nucleolus, and associated with cell proliferation and survival. In the present study, we show that EBP1 binds directly to several PPIns via two distinct PPIn-binding sites consisting of clusters of lysine residues and positioned at the N- and C-termini of the protein. Using interaction mutants, we show that the C-terminal PPIn-binding motif contributes the most to the localization of EBP1 in the nucleolus. Importantly, a K372N point mutation, located within the C-terminal motif and found in endometrial tumours, is sufficient to alter the nucleolar targeting of EBP1. Our study reveals also the presence of the class I phosphoinositide 3-kinase (PI3K) catalytic subunit p110β and its product PtdIns(3,4,5)P3 together with EBP1 in the nucleolus. Using NMR, we further demonstrate an association between EBP1 and PtdIns(3,4,5)P3 via both electrostatic and hydrophobic interactions. Taken together, these results show that EBP1 interacts directly with PPIns and associate with PtdIns(3,4,5)P3 in the nucleolus. The presence of p110β and PtdIns(3,4,5)P3 in the nucleolus indicates their potential role in regulating nucleolar processes, at least via EBP1.


Use of experiential methods to teach research in a pre-registration nursing curriculum.
For nursing to become firmly established as a research-based profession, it is essential that all nurses are equipped with the skills to critique research literature. This demands the integration of research application into nursing curricula at all levels. In order for nursing problems to be effectively illuminated, it is important that a range of quantitative and qualitative approaches are utilised. This necessitates teaching material from across the range of relevant research paradigms. Many students commencing pre-registration courses have a limited knowledge of either nursing or research. Others have had a variety of previous experiences of research as a subject, some of which have generated negative stereotypes and disinterest. This paper will explore the challenges encountered when teaching research to students on a pre-registration advanced diploma course. The authors have developed a range of experiential exercises which attempt to explore research concepts in a meaningful way. The presentation of research principles is followed up by formative work linked to clinical placements. Discussion during feedback sessions allows for sharing of ideas and experiences within the group and facilitates critical reflection. The evaluation from this initiative has been positive and encouraging. A selection of the methods used and evaluation material will be presented.


[Possible involvement of caspase activation in nitric oxide-induced neuronal apoptosis in SH-SY5Y cells].
We here report involvement of caspases in NO-induced neuronal apoptosis. Our experiments were designed to elucidate how NO induces neuronal cell death using NOC18, a new type of NO donor that spontaneously releases NO alone, without enzymatic metabolization. NOC18 induced apoptosis in human neuroblastoma SH-SY5Y cells in a concentration- and time-dependent manner estimated with DNA fragmentation assay, FACScan analysis, and nuclear morphology. In this study, oxyhemoglobin, an NO trapper, suppressed NOC18-triggered DNA fragmentation, indicating that NO from NOC18 is an apoptosis-inducer. An increase in caspase-3-like protease activity was observed in parallel with the induction of apoptosis, but no caspase-1-like protease activity was detected. The level of pro-caspase-2 protein, a precursor of caspase-2, was decreased dramatically. In addition, NOC18 also caused the cleavage of PARP, yielding an 85 kDa protein, a typical fragment of the caspases reaction. Oxyhemoglobin blocked the decrease in pro-caspase-2 and the cleavage of PARP by NOC18. Moreover, NO elicited the release of cytochrome c into the cytosol from mitochondria during apoptosis. These results suggest that activation of caspases by cytochrome c released from mitochondria is involved in neuronal apoptosis induced by NO.


Increasing prevalence of male homosexual partnerships and practices in Britain 1990-2000: evidence from national probability surveys.
To estimate the prevalence and timing of homosexual experience among British men; to explore the patterns of sexual practices and partnerships in 2000, and behavioural and attitudinal changes between 1990 and 2000 among men who have sex with men (MSM). Two large, stratified probability sample surveys of the general population. Trained interviewers administered a combination of face-to-face and self-completion questionnaires to men aged 16 to 44 years resident in Britain (n = 6000 in 1990 and n = 4762 in 2000). In 2000, 2.8% of British men reported sex with men in the past 5 years. 46.0% of MSM reported five or more partners in the past 5 years, and 59.8% reported unprotected anal intercourse in the past year. A total of 33.0% of MSM reported one or more female partner(s) in the past year. In comparison with 1990, there was a significant increase in the proportion of MSM in the population in 2000, and among these men, in the proportion reporting receptive anal intercourse in the past year [age-adjusted odds ratio (OR), 2.08; 95% confidence interval (CI), 1.08-4.00], but no significant change in self-perceived HIV-risk (age-adjusted OR, 1.11; 95% CI, 0.49-2.51) or HIV testing in past 5 years (age-adjusted OR, 1.14; 95% CI, 0.57-2.25). Evidence of increasing prevalence of homosexual intercourse among the British male population coupled with increases in some HIV-risk behaviours among MSM suggests overall increasing numbers at risk in the population. Although these changes may partly reflect an increased willingness to report these behaviours, our results are consistent with increasing incidence of sexually transmitted infections and behavioural surveillance data.


Off-pump coronary artery bypass grafting: single center experience with 1,000 consecutive patients.
Advances in surgical techniques and retractor-stabilizer devices allowing access to all coronary segments have resulted in increased interest in off-pump coronary artery bypass. The residual motion in the anastomotic site and potential hemodynamic derangements, however, render this operation technically more demanding. To evaluate the OPCAB experience in a single Israeli center. Between 2000 and 2003 in our institution, 1,000 patients underwent off-pump operations. Patients were grouped by the type of procedure, i.e., minimally invasive direct coronary artery bypass or mid-sternotomy OPCAB. One hundred MIDCAB operations were performed. Of the 900 OPCAB, 767 patients received multiple grafts with an average of 2.6 +/- 0.6 grafts per patient (range 2-4) and the remaining patients underwent single grafting during hybrid or emergency procedures. In the multiple-graft OPCAB group, complete revascularization was achieved in 96%. Multiple arterial conduits were used in 76% of the patients, and total arterial revascularization without aortic manipulation, using T-graft (35%) or in situ configurations, was performed in 61%. The respective rates for early mortality, myocardial infarction and stroke in the MIDCAB were 1%, 0% and 2%, and 2%, 1.3% and 0.9% in the multiple-vessel OPCAB groups. Multivariate analysis identified renal dysfunction (odds ratio 11.5, confidence interval 3.02-43.8; P < 0.0001) and emergency operation (OR 8.74, CL 1.99-38.3; P = 0.004) as predictors of mortality. The proportion of off-pump procedures increased from 9% prior to the study period to 59%. The use of OPCAB does not compromise the ability to achieve complete myocardial revascularization. Our procedure of choice is OPCAB using arterial conduits, preferably the 'no-touch' aorta technique.


Maternal and child access to care and intensity of conflict in the occupied Palestinian territory: a pseudo longitudinal analysis (2000-2014).
In the occupied Palestinian territory (oPt), access to maternal and child healthcare (MCH) services are constrained due to the prolonged Israeli military occupation, the Separation Wall, army checkpoints, and restrictions on the movement of people and goods. This study assesses the relationship between conflict intensity and access to Maternal and Child Health care in occupied Palestinian territory (oPt). To the best of our knowledge, the impact of conflict on access to health care has not been measured due to the lack of data. We analyse pooled data from household surveys covering a fifteen-year period (2000-2014) of children (n = 16,793) and women (n = 8477) in five regions of the oPt. Conflict intensity was used as a continuous variable defined as the square root of non-combatant conflict mortality taken from monthly death rates of non-combatants by region. We use multilevel logistic models to explain four outputs: child vaccination schedules, antenatal care, caesarean sections, and complications during pregnancy. Locality is important with results showing the negative impact of conflict intensity on access to care, especially in the South West Bank for maternal health services and Central West Bank for vaccination (B - 0.161 p = 0.000 for DPT). Wealth is only significant for DPT vaccinations with poorest (B - 0.098 p = 0.005) and poor (B - 0.148 p = 0.002) individuals less likely to access services. Otherwise conflict does not show a differential effect across socio-economic conditions. This study shows how locality is the strongest factor when looking at the impact of conflict in the oPt. Preventative services (ANC and vaccinations) are the most affected by conflict. We recommend a greater use of community health care to improve access to maternal and child care when barriers impede access to health facilities during times of conflict.


Protocol for the comparison of triflusal and clopidogrel in secondary prevention of stroke based on cytochrome P450 2C19 genotyping (MASETRO study): A multicenter, randomized, open-label, parallel-group trial.
The antiplatelet effect of clopidogrel is reportedly influenced by cytochrome P450 2C19 (CYP2C19) polymorphisms. However, there is no data concerning the relationship between stroke recurrence and CYP2C19 polymorphisms in patients treated with clopidogrel for secondary prevention of ischemic stroke. Triflusal may be an alternative therapy for clopidogrel in patients with poor genotype. The Comparison of Triflusal and Clopidogrel Effects in Secondary Prevention of Stroke Based on Cytochrome P450 2C19 Genotyping (MAESTRO) study will investigate the effect of antiplatelet agents based on CYP2C19 polymorphisms in secondary prevention of ischemic stroke. Assuming that 55% of patients belong to the poor genotype group, the required sample size is 1080 patients with at least 24 months of follow-up. This study is designed as a prospective, multicenter, randomized, parallel-group, open-label, and blind genotype trial. Patients who experience their first non-cardiogenic ischemic stroke within 30 days prior to screening are eligible. Patients received 300 mg triflusal twice a day or 75 mg clopidogrel once daily during the trial. The study is registered with ClinicalTrials.gov (NCT01174693). The primary outcome is recurrent ischemic stroke or hemorrhagic stroke. Secondary outcomes consist of composite major vascular events including stroke, myocardial infarction, coronary revascularization, or vascular death. Personalized medicine may be essential for patients according to individual drug metabolism abilities. MAESTRO is the first prospective study designed to evaluate the effect of CYP2C19 polymorphism in secondary stroke prevention and will resolve several questions regarding preventive antiplatelet agents for recurrent stroke.


Isolation, Detection, and Quantification of Cancer Biomarkers in HPV-Associated Malignancies.
Human Papillomavirus (HPV) infection has been recognized as the main etiologic factor in the development of various cancers including penile, vulva, oropharyngeal and cervical cancers. In the development of cancer, persistent HPV infections induce E6 and E7 oncoproteins, which promote cell proliferation and carcinogenesis resulting elevated levels of host antibodies (e.g., anti-HPV16 E7 antibody). Currently, these cancers are clinically diagnosed using invasive biopsy-based tests, which are performed only in centralized labs by experienced clinical staff using time-consuming and expensive tools and technologies. Therefore, these obstacles constrain their utilization at primary care clinics and in remote settings, where resources are limited. Here, we present a rapid, inexpensive, reliable, easy-to-use, customized immunoassay platform following a microfluidic filter device to detect and quantify anti-HPV16 E7 antibodies from whole blood as a non-invasive assisting technology for diagnosis of HPV-associated malignancies, especially, at primary healthcare and remote settings. The platform can detect and quantify anti-HPV16 E7 antibody down to 2.87 ng/mL. We further validated our immunoassay in clinical patient samples and it provided significantly high responses as compared to control samples. Thus, it can be potentially implemented as a pretesting tool to identify high-risk groups for broad monitoring of HPV-associated cancers in resource-constrained settings.


Human-centric sensing.
The first decade of the century witnessed a proliferation of devices with sensing and communication capabilities in the possession of the average individual. Examples range from camera phones and wireless global positioning system units to sensor-equipped, networked fitness devices and entertainment platforms (such as Wii). Social networking platforms emerged, such as Twitter, that allow sharing information in real time. The unprecedented deployment scale of such sensors and connectivity options ushers in an era of novel data-driven applications that rely on inputs collected by networks of humans or measured by sensors acting on their behalf. These applications will impact domains as diverse as health, transportation, energy, disaster recovery, intelligence and warfare. This paper surveys the important opportunities in human-centric sensing, identifies challenges brought about by such opportunities and describes emerging solutions to these challenges.


Role of somatostatin in the prevention of pancreatic stump-related morbidity following elective pancreaticoduodenectomy in high-risk patients and elimination of surgeon-related factors: prospective, randomized, controlled trial.
A prospective, randomized, controlled trial was performed to determine the efficacy of somatostatin in the prevention of pancreatic stump-related complications with elimination of surgeon-related factors in high-risk patients undergoing pancreaticoduodenectomy. From August 1997 to December 2000, 54 patients, 28 men and 26 women, with age ranged from 32 to 89 years, were randomly assigned to somatostatin group ( n = 27) or placebo group ( n = 27). Ninety-four percent of the patients had pancreatic and periampullary lesions; 6% had secondary lesion involving the duodenum such as local recurrent colon carcinoma and renal cell carcinoma. These patients received either standard pancreaticoduodenectomy or pylorus-preserving pancreaticoduodenectomy. An experienced surgeon performed all operations in same fashion to minimize the surgical factor. A transanastomotic tube was inserted into the pancreatic duct for diversion of pancreatic juice in the pancreaticojejunostomy for a 3-weeks period postoperatively. Intravenous infusion of somatostatin was given at a dose of 250 microg/hr in the somastotatin group and normal saline was given to the control group for 7 days postoperatively. There was one perioperative death in each group, resulting in a 3.7% mortality rate. In the somastotatin group, as compared to the placebo group, the incidence of overall morbidity and pancreatic stump related complications were significantly lower with a mean decrease of 50% pancreatic juice output and a slightly shorter duration of hospital stays. In conclusion, after excluding surgeon related factor, prophylactic use of somatostatin reduces the incidence and severity of pancreatic stump related complications in high-risk patients having pancreaticoduodenectomy via decreased secretion of pancreatic exocrine.


Mode of Commuting to School and Its Association with Physical Activity and Sedentary Habits in Young Ecuadorian Students.
Active commuting to and from school (ACS) could help to increase daily physical activity levels in youth; however, this association remains unknown in Ecuadorian youth. Thus, the aims of this study were (1) to determine the patterns of commuting to and from school and (2) to analyze the associations between ACS, physical activity (PA), and sedentary habits in Ecuadorian youth. A total of 732 students (65.3% males), aged 10⁻18 years (children = 246, young adolescents = 310, older adolescents = 162) from the central region of Ecuador participated in this study. A self-report questionnaire, including the usual mode and frequency of commuting, distance from home to school (PACO-Questionnaire), and PA and sedentary habits (YAP-Questionnaire), was used. Most of the sample lived ≤2 km from school; however, they were mainly passive commuters (96%). The most common mode of commuting was by car (to school = 43.4%, from school = 31.6%; p < 0.001). Children presented significantly higher scores (0⁻4) in PA outside school and total PA compared with older adolescents (2.20 ± 0.97 vs. 1.97 ± 0.96; p = 0.013 and 2.30 ± 0.76 vs. 2.09 ± 0.74, p = 0.019, respectively), as well as the lowest scores in sedentary habits (1.51 ± 0.65, p < 0.001). PA at school and total PA were positively associated with ACS (OR 3.137; 95% CI, 1.918 to 5.131; p < 0.001, and OR 2.543; 95% CI, 1.428 to 4.527; p = 0.002, respectively). In conclusion, passive modes of transportation were the most frequently used to commute to and from school in young Ecuadorians. PA at school and total PA were positively associated with ACS. Thus, interventions at school setting could be an opportunity to improve PA levels and additionally ACS in youth from the central region of Ecuador.


Racial Disparities in End-of-Life Care Among Patients With Prostate Cancer: A Population-Based Study.
To examine racial disparities in end-of-life (EOL) care among black and white patients dying of prostate cancer (PCa). Relying on the SEER-Medicare database, 3789 patients who died of metastatic PCa between 1999 and 2009 were identified. Information was assessed regarding diagnostic care, therapeutic interventions, hospitalizations, intensive care unit (ICU) admissions, and emergency department visits in the last 12 months, 3 months, and 1 month of life. Logistic regression tested the relationship between race and the receipt of diagnostic care, therapeutic interventions, and high-intensity EOL care. Overall, 729 patients (19.24%) were black. In the 12-months preceding death, laboratory tests (odds ratio [OR], 0.51; 95% CI, 0.36-0.72), prostate-specific antigen test (OR, 0.54; 95% CI, 0.43-0.67), cystourethroscopy (OR, 0.71; 95% CI, 0.56-0.90), imaging procedure (OR, 0.58; 95% CI, 0.41-0.81), hormonal therapy (OR, 0.53; 95% CI, 0.44-0.65), chemotherapy (OR, 0.59; 95% CI, 0.48-0.72), radiotherapy (OR, 0.74; 95% CI, 0.61-0.90), and office visit (OR, 0.38; 95% CI, 0.28-0.50) were less frequent in black versus white patients. Conversely, high-intensity EOL care, such as ICU admission (OR, 1.27; 95% CI, 1.04-1.58), inpatient admission (OR, 1.49; 95% CI, 1.09-2.05), and cardiopulmonary resuscitation (OR, 1.72; 95% CI, 1.40-2.11), was more frequent in black versus white patients. Similar trends for EOL care were observed at 3-month and 1-month end points. Although diagnostic and therapeutic interventions are less frequent in black patients with end-stage PCa, the rate of high-intensity and aggressive EOL care is higher in these individuals. These disparities may indicate that race plays an important role in the quality of care for men with end-stage PCa.


PB1-F2 proteins from H5N1 and 20 century pandemic influenza viruses cause immunopathology.
With the recent emergence of a novel pandemic strain, there is presently intense interest in understanding the molecular signatures of virulence of influenza viruses. PB1-F2 proteins from epidemiologically important influenza A virus strains were studied to determine their function and contribution to virulence. Using 27-mer peptides derived from the C-terminal sequence of PB1-F2 and chimeric viruses engineered on a common background, we demonstrated that induction of cell death through PB1-F2 is dependent upon BAK/BAX mediated cytochrome c release from mitochondria. This function was specific for the PB1-F2 protein of A/Puerto Rico/8/34 and was not seen using PB1-F2 peptides derived from past pandemic strains. However, PB1-F2 proteins from the three pandemic strains of the 20(th) century and a highly pathogenic strain of the H5N1 subtype were shown to enhance the lung inflammatory response resulting in increased pathology. Recently circulating seasonal influenza A strains were not capable of this pro-inflammatory function, having lost the PB1-F2 protein's immunostimulatory activity through truncation or mutation during adaptation in humans. These data suggest that the PB1-F2 protein contributes to the virulence of pandemic strains when the PB1 gene segment is recently derived from the avian reservoir.


Alkalinity and formation of zeolites in saline alkaline lakes.
The solubility of rhyolitic glass increases with increasing alkalinity, whereas the ratio of silicon to aluminum decreases with increasing alkalinity. The strong correlation observed between alkalinity and zeolite mineralogy in saline, alkaline lakes is thought to be a function of this relationship between pH and the Si/Al ratio. It is suggested that this function is a result of the reaction between silicic glass and alkaline solution whereby (i) a gel forms, whose Si/Al ratio is controlled by the Si/Al ratio of the solution, and (ii) a zeolite forms from the gel, whose Si/Al ratio is, in turn, controlled by the composition of the gel.


An integrated approach to the biomechanics and motor control of cricket fast bowling techniques.
To date, scientific investigations into the biomechanical aspects of cricket fast bowling techniques have predominantly focused on identifying the mechanical factors that may predispose fast bowlers to lower back injury with a relative paucity of research being conducted on the technical features that underpin proficient fast bowling performance. In this review paper, we critique the scientific literature examining fast bowling performance. We argue that, although many published investigations have provided some useful insights into the biomechanical factors that contribute to a high ball release speed and, to a lesser extent, bowling accuracy, this research has not made a substantive contribution to knowledge enhancement and has only had a very minor influence on coaching practice. To significantly enhance understanding of cricket fast bowling techniques and, therefore, have greater impact on practice, we recommend that future scientific research adopts an interdisciplinary focus, integrating biomechanical measurements with the analytical tools and concepts of dynamical systems motor control theory. The use of qualitative (topological) analysis techniques, in particular, promises to increase understanding of the coordinative movement patterns that define 'technique' in cricket fast bowling and potentially help distinguish between functional and dysfunctional aspects of technique for individual fast bowlers.


Regulation of endocytosis by CUP-5, the Caenorhabditis elegans mucolipin-1 homolog.
Loss of the human mucolipin-1 gene underlies mucolipidosis type IV (MLIV), a lysosomal storage disease that results in severe developmental neuropathology. Unlike other lysosomal storage diseases, MLIV is not associated with a lack of lysosomal hydrolases; instead, MLIV cells display abnormal endocytosis of lipids and accumulate large vesicles, indicating that a defect in endocytosis may underlie the disease. Here we report the identification of a loss-of-function mutation in the Caenorhabditis elegans mucolipin-1 homolog, cup-5, and show that this mutation results in an enhanced rate of uptake of fluid-phase markers, decreased degradation of endocytosed protein and accumulation of large vacuoles. Overexpression of cup-5(+) causes the opposite phenotype, indicating that cup-5 activity controls aspects of endocytosis. Studies in model organisms such as C. elegans have helped illuminate fundamental mechanisms involved in normal cellular function and human disease; thus the C. elegans cup-5 mutant may be a useful model for studying conserved aspects of mucolipin-1 structure and function and for assessing the effects of potential therapeutic compounds.


Therapeutic interventions for spinal muscular atrophy: preclinical and early clinical development opportunities.
Spinal muscular atrophy (SMA) is an autosomal recessive neurodegenerative neuromuscular disease that presents primarily in children. Abnormalities in the SMN1 gene cause reduced levels of the survival motor neuron (SMN) protein, while a second gene, SMN2, produces low levels of functional SMN protein. Currently available drugs do not cure, so a significant unmet need remains for patients treated after symptom onset. Drugs available in the clinic, investigational agents and key questions for researchers are discussed. A pragmatic search of the literature was performed to identify therapies in late stages of preclinical, or in early stages of clinical development. This list was compared to the CureSMA pipeline for completeness. Drugs approved for indications that have potential for impact for SMA were included. These drugs target the primary deficiency in SMN protein or other pathways involved in SMA pathophysiology that are not SMN-protein dependent. Children treated after the onset of symptoms continue to have significant disability. Given the heterogeneity of the population phenotype evidenced by variable response to initial therapy, age at treatment onset and the need to demonstrate added value beyond approved therapeutics, the clinical development of new drugs will be challenging.


Label-free proteomic analysis of breast cancer molecular subtypes.
To better characterize the cellular pathways involved in breast cancer molecular subtypes, we performed a proteomic study using a label-free LC-MS strategy for determining the proteomic profile of Luminal A, Luminal-HER2, HER2-positive, and triple-negative (TN) breast tumors compared with healthy mammary tissue. This comparison aimed to identify the aberrant processes specific for each subtype and might help to refine our understanding regarding breast cancer biology. Our results address important molecular features (both specific and commonly shared) that explain the biological behavior of each subtype. Changes in proteins related to cytoskeletal organization were found in all tumor subtypes, indicating that breast tumors are under constant structural modifications to invade and metastasize. We also found changes in cell-adhesion processes in all molecular subtypes, corroborating that invasiveness is a common property of breast cancer cells. Luminal-HER2 and HER2 tumors also presented altered cell cycle regulation, as shown by the several DNA repair-related proteins. An altered immune response was also found as a common process in the Luminal A, Luminal-HER2, and TN subtypes, and complement was the most important pathway. Analysis of the TN subtype revealed blood coagulation as the most relevant biological process.


The effects of tissue factor pathway inhibitor and anti-beta-2-glycoprotein-I IgG on thrombin generation.
Recent evidence suggests that autoantibodies to tissue factor pathway inhibitor (TFPI) and/or antiphospholipid antibodies (aPL) may contribute to upregulation of the tissue factor (TF) pathway of blood coagulation and the development of thrombotic complications in the antiphospholipid syndrome (aPS). The aim of this study was to determine the influence of aPL e.g. anti-beta-2-glycoprotein-I (anti-beta2GPI) and anti-prothrombin, on in vitro TF-induced thrombin generation in the presence and absence of TFPI. IgG fractions were collected from subjects with aPL (n=21) and normal controls (n=36). Anti-TFPI activity was determined after incubation of IgG isolated from control or subject plasma with pooled normal plasma using an amidolytic assay for TFPI. The influence of IgG fractions and purified aPL (anti-beta2GPI and anti-prothrombin) on TF-induced in vitro thrombin generation was determined using a chromogenic assay of thrombin activity. Patients with aPL had significantly elevated thrombin generation (median [interquartile range]) compared to normal controls (112.0 [104.0-124.0]% vs 89.9 [85.7-100.9]%, respectively; p<0.001). Thrombin generation was significantly correlated with anti-TFPI activity in patients with aPL (r(s)=0.452; p=0.039). It was also demonstrated that anti-beta2GPI, but not anti-prothrombin IgG antibodies, significantly enhanced TF-induced thrombin generation in the presence of TFPI, using both purified and patients' samples. Our findings support the hypothesis that anti-beta2GPI IgG antibodies accelerate thrombin generation in the presence of TFPI and may contribute to hypercoagulability in patients with aPS.


Practical prevention of cardiac remodeling and atrial fibrillation with full-spectrum antioxidant therapy and ancillary strategies.
A wealth of research data points to increased oxidative stress as a key driver of the cardiac remodeling triggered by chronic pressure overload, loss of functional myocardial tissue, or atrial fibrillation. Oxidative stress is a mediator of the cardiomyocyte hypertrophy and apoptosis, the cardiac fibrosis, and the deficits in cardiac function which typify this syndrome, and may play a role in initiating and sustaining atrial fibrillation. Nox2- and Nox4-dependent NADPH oxidase activity appears to be a major source of this oxidative stress, and oxidants can induce conformational changes in xanthine dehydrogenase, nitric oxide synthase, and the mitochondrial respiratory chain which increase their capacity to generate superoxide as well. Consistent with these insights, various synthetic antioxidants have been shown to suppress cardiac remodeling in rodents subjected to myocardial infarction, aortic constriction, or rapid atrial pacing. It may prove feasible to achieve comparable benefits in humans through use of a "full-spectrum antioxidant therapy" (FSAT) that features a complementary array of natural antioxidants. Spirulina is a rich source of phycocyanobilin, a derivative and homolog of biliverdin that appears to mimic the potent inhibitory impact of biliverdin and free bilirubin on NADPH oxidase activity. Mega-doses of folate can markedly increase intracellular levels of tetrahydrofolates which have potent and versatile radical-scavenging activities - including efficient quenching of peroxynitrite-derived radicals Supplemental coenzyme Q10, already shown to improve heart function in clinical congestive failure, can provide important antioxidant protection to mitochondria. Phase 2 inducer nutraceuticals such as lipoic acid, administered in conjunction with N-acetylcysteine, have the potential to blunt the impact of oxidative stress by boosting myocardial levels of glutathione. While taurine can function as an antioxidant for myeloperoxidase-derived radicals, its positive inotropic effect on the failing heart seems more likely to reflect an effect on intracellular calcium dynamics. These measures could aid control of cardiac modeling less directly by lowering elevated blood pressure, or by aiding the perfusion of ischemic cardiac regions through an improvement in coronary endothelial function. Since nitric oxide functions physiologically to oppose cardiomyocyte hypertrophy and cardiac fibrosis, and is also a key regulator of blood pressure and endothelial function, cocoa flavanols - which provoke endothelial release of nitric oxide - might usefully complement the antioxidant measures recommended here.


Influence of phlebographic abnormalities on the natural history of venous ulceration.
Eighty-five limbs in 73 patients with a healed venous ulcer were assessed by ascending and descending phlebography, foot volume plethysmography and transcutaneous oxygen measurements. Forty-four limbs had post-thrombotic changes on ascending phlebography. In 24 (28 per cent) these extended into the femoral vein, while in 20 (24 per cent) only the calf veins were involved. In the 41 limbs (48 per cent) with normal deep veins on ascending phlebography, 11 had evidence of localized incompetence of the calf communicating veins, 14 had either long saphenous incompetence, deep vein reflux to the level of the knee or below, or both of these abnormalities, and 16 limbs had no phlebographic abnormalities. However all limbs had a decreased half volume refilling time on foot volume plethysmography. Limbs with post-thrombotic changes extending into the femoral vein were associated with a significantly longer history of ulceration and more ulcer recurrences than limbs with calf vein damage (P less than 0.05 for each) and limbs with normal deep veins (P less than 0.01 for each). However, these limbs did not have lower transcutaneous oxygen ratios or longer times to achieve ulcer healing. Ascending phlebography identified a group of limbs with extensive post-thrombotic changes in which there was a higher incidence of ulcer breakdown, but this was not associated with a delay in ulcer healing.


Shaking up glycolysis: Sustained, high lactate flux during aerobic rattling.
Substantial ATP supply by glycolysis is thought to reflect cellular anoxia in vertebrate muscle. An alternative hypothesis is that the lactate generated during contraction reflects sustained glycolytic ATP supply under well-oxygenated conditions. We distinguished these hypotheses by comparing intracellular glycolysis during anoxia to lactate efflux from muscle during sustained, aerobic contractions. We examined the tailshaker muscle of the rattlesnake because of its uniform cell properties, exclusive blood circulation, and ability to sustain rattling for prolonged periods. Here we show that glycolysis is independent of the O(2) level and supplies one-third of the high ATP demands of sustained tailshaking. Fatigue is avoided by rapid H(+) and lactate efflux resulting from blood flow rates that are among the highest reported for vertebrate muscle. These results reject the hypothesis that glycolysis necessarily reflects cellular anoxia. Instead, they demonstrate that glycolysis can provide a high and sustainable supply of ATP along with oxidative phosphorylation without muscle fatigue.


Glyphosate-rich air samples induce IL-33, TSLP and generate IL-13 dependent airway inflammation.
Several low weight molecules have often been implicated in the induction of occupational asthma. Glyphosate, a small molecule herbicide, is widely used in the world. There is a controversy regarding a role of glyphosate in developing asthma and rhinitis among farmers, the mechanism of which is unexplored. The aim of this study was to explore the mechanisms of glyphosate induced pulmonary pathology by utilizing murine models and real environmental samples. C57BL/6, TLR4-/-, and IL-13-/- mice inhaled extracts of glyphosate-rich air samples collected on farms during spraying of herbicides or inhaled different doses of glyphosate and ovalbumin. The cellular response, humoral response, and lung function of exposed mice were evaluated. Exposure to glyphosate-rich air samples as well as glyphosate alone to the lungs increased: eosinophil and neutrophil counts, mast cell degranulation, and production of IL-33, TSLP, IL-13, and IL-5. In contrast, in vivo systemic IL-4 production was not increased. Co-administration of ovalbumin with glyphosate did not substantially change the inflammatory immune response. However, IL-13-deficiency resulted in diminished inflammatory response but did not have a significant effect on airway resistance upon methacholine challenge after 7 or 21 days of glyphosate exposure. Glyphosate-rich farm air samples as well as glyphosate alone were found to induce pulmonary IL-13-dependent inflammation and promote Th2 type cytokines, but not IL-4 for glyphosate alone. This study, for the first time, provides evidence for the mechanism of glyphosate-induced occupational lung disease.


Changes in Midline Tremor and Gait Following Deep Brain Stimulation for Essential Tremor.
Essential tremor (ET) is a common movement disorder characterized by kinetic and postural tremor in the upper extremities and frequently in the midline. Persons with ET often also exhibit gait ataxia. Previous studies have observed associations between midline tremor severity and gait ataxia in persons with ET, suggesting a common pathophysiology distinct from that of upper extremity tremor. However, a causal link between midline tremor and gait impairment has not been established. We investigated tremor and gait in 24 persons with ET before and after implantation of unilateral deep brain stimulation into the ventralis intermedius nucleus of the thalamus. Stimulation significantly improved tremor in the targeted upper extremity and midline. However, gait was unaffected at the cohort level. Furthermore, improvement in midline tremor was not significantly associated with gait improvement. These findings revealed that midline tremor and gait impairment may be dissociable in persons with ET.


The earliest settlers' antiquity and evolutionary history of Indian populations: evidence from M2 mtDNA lineage.
The "out of Africa" model postulating single "southern route" dispersal posits arrival of "Anatomically Modern Human" to Indian subcontinent around 66-70 thousand years before present (kyBP). However the contributions and legacy of these earliest settlers in contemporary Indian populations, owing to the complex past population dynamics and later migrations has been an issue of controversy. The high frequency of mitochondrial lineage "M2" consistent with its greater age and distribution suggests that it may represent the phylogenetic signature of earliest settlers. Accordingly, we attempted to re-evaluate the impact and contribution of earliest settlers in shaping the genetic diversity and structure of contemporary Indian populations; using our newly sequenced 72 and 4 published complete mitochondrial genomes of this lineage. The M2 lineage, harbouring two deep rooting subclades M2a and M2b encompasses approximately one tenth of the mtDNA pool of studied tribes. The phylogeographic spread and diversity indices of M2 and its subclades among the tribes of different geographic regions and linguistic phyla were investigated in detail. Further the reconstructed demographic history of M2 lineage as a surrogate of earliest settlers' component revealed that the demographic events with pronounced regional variations had played pivotal role in shaping the complex net of populations phylogenetic relationship in Indian subcontinent. Our results suggest that tribes of southern and eastern region along with Dravidian and Austro-Asiatic speakers of central India are the modern representatives of earliest settlers of subcontinent. The Last Glacial Maximum aridity and post LGM population growth mechanised some sort of homogeneity and redistribution of earliest settlers' component in India. The demic diffusion of agriculture and associated technologies around 3 kyBP, which might have marginalized hunter-gatherer, is coincidental with the decline of earliest settlers' population during this period.


Management of intersection syndrome in professional motorcycle rider: a case report.
Intersection syndrome is an uncommon and underdiagnosed condition, especially found in sport overuse. This case report describes the treatment of a professional motor biker who presented intense wrist pain and swelling after training sessions. The athlete was treated with rest, the use of a wrist splint combined with NSAIDs, physical therapy, including pulsed Nd-YAG laser therapy and exercise, such as stretching and release, massage and eccentric training. The applied therapeutic protocol was successful and allowed the biker to completely resolve the symptoms and return to training session and professional races at full power. The results are durable at 5 months post injury follow up. This experience suggests that pulsed Nd-YAG laser therapy can be an additional useful tool in the global approach treatment for intersection syndrome, contributing with splint and manual therapy, to full recovery even in mechanically stressful conditions. (www.actabiomedica.it).


The relationship between odour annoyance scores and modelled ambient air pollution in Sarnia, "Chemical Valley", Ontario.
This study aimed at establishing the relationship between annoyance scores and modelled air pollution in "Chemical Valley", Sarnia, Ontario (Canada). Annoyance scores were taken from a community health survey (N = 774); and respondents' exposure to nitrogen dioxide (NO(2)) and sulphur dioxide (SO(2)) were estimated using land use regression (LUR) models. The associations were examined by univariate analysis while multivariate logistic regression was used to examine the determinants of odour annoyance. The results showed that odour annoyance was significantly correlated to modelled pollutants at the individual (NO(2), r = 0.15; SO(2), r = 0.13) and census tract (NO(2), r = 0.56; SO(2), r = 0.67) levels. The exposure-response relationships show that residents of Sarnia react to very low pollution concentrations levels even if they are within the Ontario ambient air quality criteria. The study found that exposure to high NO(2) and SO(2) concentrations, gender, and perception of health effects were significant determinants of individual odour annoyance reporting. The observed association between odour annoyance and modelled ambient pollution suggest that individual and census tract level annoyance scores may serve as proxies for air quality in exposed communities because they capture the within area spatial variability of pollution. However, questionnaire-based odour annoyance scores need to be validated longitudinally and across different scales if they are to be adopted for use at the national level.


How do basophils contribute to Th2 cell differentiation and allergic responses?
Basophils and mast cells share some features, including basophilic granules in the cytoplasm, cell surface expression of the high-affinity IgE receptor and release of chemical mediators such as histamine. Because of this similarity and their minority status, basophils had often been erroneously considered as minor relatives or blood-circulating precursors of tissue-resident mast cells, and therefore long been neglected or underestimated in immunological studies. Taking advantage of newly developed tools, such as basophil-depleting antibodies and engineered mice deficient for only basophils, recent studies have identified previously unappreciated roles for basophils, distinct from those played by mast cells, in allergic responses, protective immunity against parasitic infections and regulation of other immune cells. In this review, we focus on two topics that we presented and discussed in the 46th Annual Meeting of the Japanese Society for Immunology held in Sendai in December 2017. The first topic is the function of basophils as antigen-presenting cells for driving Th2 cell differentiation. We demonstrated that basophils produce few or no MHC class II (MHC-II) proteins by themselves although they can acquire peptide-MHC-II complexes from dendritic cells through trogocytosis, and present them and provide IL-4 to naive CD4 T cells, promoting Th2 cell differentiation. The second topic is the basophil-specific effector molecules involved in allergic responses. Among mouse mast cell proteases (mMCPs), mMCP-8 and mMCP-11 are expressed almost exclusively by basophils. Analyses in vitro and in vivo revealed that both proteases can induce leukocyte migration through distinct mechanisms, contributing to the development of basophil-dependent allergic inflammation.


Fluidic delivery of homogeneous solutions through carbon tube bundles.
A wide array of technological applications requires localized high-rate delivery of dissolved compounds (in particular, biological ones), which can be achieved by forcing the solutions or suspensions of such compounds through nano or microtubes and their bundled assemblies. Using a water-soluble compound, the fluorescent dye Rhodamine 610 chloride, frequently used as a model drug release compound, it is shown that deposit buildup on the inner walls of the delivery channels and its adverse consequences pose a severe challenge to implementing pressure-driven long-term fluidic delivery through nano and microcapillaries, even in the case of such homogeneous solutions. Pressure-driven delivery (3-6 bar) of homogeneous dye solutions through macroscopically-long (approximately 1 cm) carbon nano and microtubes with inner diameters in the range 100 nm-1 microm and their bundled parallel assemblies is studied experimentally and theoretically. It is shown that the flow delivery gradually shifts from fast convection-dominated (unobstructed) to slow jammed convection, and ultimately to diffusion-limited transport through a porous deposit. The jamming/clogging phenomena appear to be rather generic: they were observed in a wide concentration range for two fluorescent dyes in carbon nano and microtubes, as well as in comparable transparent glass microcapillaries. The aim of the present work is to study the physics of jamming, rather than the chemical reasons for the affinity of dye molecules to the tube walls.


The natural history of cervical disc calcification in children.
Calcification of cervical intervertebral discs is uncommon in children. The cases of approximately 200 children have been reported in the literature. The objective of the present study was to examine the natural history of this condition. Seventeen consecutive children, ten boys and seven girls, with calcified cervical intervertebral discs were included in the study. The mean age at diagnosis was 7.7 years. Three children had had a fever, and two had a history of trauma. All of the children complained of neck pain except one who was asymptomatic and was identified by chance. On radiographs, the calcification was seen within the central part of the cervical intervertebral discs. All children had treatment of the symptoms, and they were then followed periodically until the disappearance of both the symptoms and the calcific deposits. The children were followed for a mean of five years, and all had a complete disappearance of both the symptoms and the calcification. None had recurrent symptoms or calcifications. The average time to resolution of the symptoms and the calcifications was 34.3 days (range, seventeen to 173 days) and fifteen months (range, three to sixty months), respectively. With the numbers available, factors such as age, level of disc calcification, severity of symptoms, and degree of spinal canal compromise were not related to the time of resolution. Cervical disc calcification in children is a self-limiting condition with an excellent prognosis. Even if clinical symptoms due to nerve-root or spinal cord compression are present, conservative therapy produces satisfactory results. Therefore, nerve-root or spinal cord compression by a calcified cervical disc should not be considered an absolute indication for surgical intervention.


Exploiting the terminological approach from CEN/TC251 and GALEN to support semantic interoperability of healthcare record systems.
We apply the principles included in two CEN standards (ENV 12265, ENV 12264) to the analysis of the semantic structure of health record systems, to support their semantic interoperability. This result was made possible by dramatic methodological progress in the field of terminological systems--due to a worldwide evolution towards a new generation--and by the experience we acquired in the GALEN-IN-USE (formerly GALEN) project. The meaning behind names, content and context of record items and record item complexes can be considered as a 'semantic continuum'. This continuum is made explicit, by building a suitable paraphrase in a controlled language. We can then apply the principles we previously elaborated for the second generation of terminological system. Methodology and tools for generating a controlled language and a second-generation terminological system were developed and successfully used in the GALEN-IN-USE project and promising experiments were performed on elements of record structure listed in LOINC and SDM. In this way, the semantic structures of different record systems can be expressed by the resulting common formalism and thus, information units can be faithfully exchanged among different structures.


Reconstructing metabolic flux vectors from extreme pathways: defining the alpha-spectrum.
The move towards genome-scale analysis of cellular functions has necessitated the development of analytical (in silico) methods to understand such large and complex biochemical reaction networks. One such method is extreme pathway analysis that uses stoichiometry and thermodynamic irreversibly to define mathematically unique, systemic metabolic pathways. These extreme pathways form the edges of a high-dimensional convex cone in the flux space that contains all the attainable steady state solutions, or flux distributions, for the metabolic network. By definition, any steady state flux distribution can be described as a nonnegative linear combination of the extreme pathways. To date, much effort has been focused on calculating, defining, and understanding these extreme pathways. However, little work has been performed to determine how these extreme pathways contribute to a given steady state flux distribution. This study represents an initial effort aimed at defining how physiological steady state solutions can be reconstructed from a network's extreme pathways. In general, there is not a unique set of nonnegative weightings on the extreme pathways that produce a given steady state flux distribution but rather a range of possible values. This range can be determined using linear optimization to maximize and minimize the weightings of a particular extreme pathway in the reconstruction, resulting in what we have termed the alpha-spectrum. The alpha-spectrum defines which extreme pathways can and cannot be included in the reconstruction of a given steady state flux distribution and to what extent they individually contribute to the reconstruction. It is shown that accounting for transcriptional regulatory constraints can considerably shrink the alpha-spectrum. The alpha-spectrum is computed and interpreted for two cases; first, optimal states of a skeleton representation of core metabolism that include transcriptional regulation, and second for human red blood cell metabolism under various physiological, non-optimal conditions.


Relieving symptoms in cancer: innovative use of art therapy.
Art therapy has been used in a variety of clinical settings and populations, although few studies have explored its use in cancer symptom control. The specific aim of this study was to determine the effect of a 1-hour art therapy session on pain and other symptoms common to adult cancer inpatients. A quasi-experimental design was used (n=50). The Edmonton Symptom Assessment Scale (ESAS) and the Spielberger State-Trait Anxiety Index (STAI-S) were used prior to and after the art therapy to quantify symptoms, while open-ended questions evaluated the subjects' perceptions of the experience. There were statistically significant reductions in eight of nine symptoms measured by the ESAS, including the global distress score, as well as significant differences in most of the domains measured by the STAI-S. Subjects overwhelmingly expressed comfort with the process and desire to continue with therapy. This study provides beginning evidence for the efficacy of art therapy in reducing a broad spectrum of symptoms in cancer inpatients.


Molecular and immunohistochemical analysis of intraductal papillary neoplasms of the biliary tract.
Intraductal papillary neoplasms (IPNs) of the biliary tract are uncommon lesions that may be solitary or may spread extensively along the biliary tree. Some biliary IPNs are histologically and radiologically similar to intraductal papillary mucinous tumors (IPMNs) of the pancreas and present a risk for progression to invasive cholangiocarcinoma. Unlike pancreatic IPMNs, little is known about their molecular pathogenesis. We studied 14 biliary IPNs (including 5 cases with associated invasive cholangiocarcinoma) for genetic alterations in the APC/beta-catenin pathway, K-ras oncogene mutations, p53/chromosome 17p alterations, and Dpc4/18q alterations. Immunohistochemistry was performed for beta-catenin, p53, and Dpc4, and microdissected tissue was analyzed using direct DNA sequencing for exon 1 of K-ras and exon 3 of beta-catenin and allelic loss assays on chromosomes 5q, 17p, and 18q. Activating mutations in codon 12 of the K-ras oncogene were present in 4 of 14 (29%) biliary IPNs. Of these 4 cases, 2 patients had associated invasive cholangiocarcinoma, and identical K-ras mutations were present in both the intraductal and invasive components. Allelic loss on chromosome 18q was present in 4 of 13 informative cases (31%); however, no loss of normal Dpc4 expression was detected by immunohistochemistry. Nuclear accumulation of beta-catenin protein was demonstrated in 3 of 12 cases (25%); however, there were no beta-catenin gene mutations, and allelic loss on 5q was present in only 1 of 10 informative cases (10%). Both immunohistochemistry for p53 and 17p allelic loss assays were negative. Biliary IPNs therefore demonstrate a K-ras gene mutation frequency that is lower than that previously reported for pancreatic IPMNs, but similar to that reported for hepatic cholangiocarcinomas. The presence of K-ras mutations in 2 purely intraductal neoplasms, and identical K-ras mutations in 2 cases with both intraductal and invasive components, suggests that these mutations arise early in tumorigenesis. Finally, the frequency of allelic loss on 18q suggests that a locus on 18q is involved in the molecular pathogenesis of biliary IPNs, but this locus is not DPC4.


Immunomodulatory properties of maxadilan, the vasodilator peptide from sand fly salivary gland extracts.
Sand flies are the arthropod vector of leishmaniasis and salivary gland extracts from these flies exacerbate leishmaniasis in vivo. The mechanism of exacerbation appears to be due to immunomodulatory effects of the saliva on host immune function but the active component is unknown. The following studies reveal that maxadilan, the vasodilatory peptide present in sand fly salivary gland extracts, has immunomodulatory properties. To examine the effect of maxadilan on T cell proliferation, the peptide was added to murine spleen cells stimulated with either concanavalin A or plate-bound anti-T cell receptor antibody. Inhibition of proliferation was noted in a dose-dependent manner for both sets of experiments (P < 0.05). To examine the effect of maxadilan on alloantigen presentation, the peptide was added to mixed lymphocyte and mixed epidermal cell lymphocyte reactions. Inhibition of proliferation was found in these culture systems. Maxadilan also inhibited the delayed-type hypersensitivity reaction in mice (P < 0.05). These observations suggest a role for maxadilan in the pathogenesis of leishmaniasis since the peptide may inhibit the immune response at the site of parasite inoculation, allowing the infection to proceed.


LIMPRINT: The UK Experience-Subjective Control of Swelling in Patients Attending Specialist Lymphedema Services.
Background and Study Design: This study was undertaken as part of the UK LIMPRINT international study to determine the number of people with chronic edema (CO) and its impact on health services. Overall 7436 with CO were recruited in the main UK study from a range of health settings. Methods and Results: Factors relating to subjective control of arm and leg CO were defined in the UK. A total of 1565 patients were included in the study with exclusions for: no limb swelling or not recorded (1669), having concurrent arm/leg CO (272), control of assessment missing (5) and professional being unsure of control status of CO (325). Arm swelling occurred in 953 (18.5%) with leg CO in 4212 (81.5%). Poor control was found in 1430 (27.2%) and good control in 3735 (72.3%). Control of arm swelling was worse in men and control increased overall in those aged over 45 years. In contrast control of CO worsened in those with leg CO with increasing age and multiple co-morbidities. Obesity and cellulitis, particularly an episode in the last year were associated with poor control. Independent risk factors for arm CO were : obesity, neurological disease and cellulitis in the last year and for leg CO, obesity, poor mobility, heart disease, presence of a wound, cellulitis in the last year and duration of swelling. Conclusion: Control of CO within specialized centers is complex due to sociodemographic and clinical comorbidities.