Back pain in athletes.
The athlete with back pain presents a clinical challenge. Self-limited symptoms must be distinguished from persistent or recurrent symptoms associated with identifiable pathology. Athletes involved in impact sports appear to have risk factors for specific spinal pathologies that correlate with the loading and repetition demands of specific activities. For example, elite athletes who participate in longer and more intense training have higher incidence rates of degenerative disk disease and spondylolysis than athletes who do not. However, data suggest that the recreational athlete may be protected from lumbar injury with physical conditioning. Treatment of athletes with acute or chronic back pain usually is nonsurgical, and symptoms generally are self-limited. However, a systematic approach to the athlete with back pain, involving a thorough history and physical examination, pertinent imaging, and treatment algorithms designed for specific diagnoses, can facilitate symptomatic improvement and return to play. There are no reliable studies examining the long-term consequences of athletic activity on the lumbar spine.


Long-life partners or sex friends? Impact of parental pair bond on offspring personality.
Previous investigations reported that some traits of parental relationships, including pair-bond duration or mate behavioural compatibility, influence subsequent offspring fitness by acting on their behaviour and growth and thus their early survival. We hypothesized that the development of a pair bond between sexual partners would have a prenatal influence. This study investigated the impact of two pairing managements on the egg characteristics and development of offspring of Japanese quail (Coturnix c. japonica). Thirty males and 30 females were paired either continuously (C; mates together all the time) or non-continuously (NC; pairs met only three times a week for 5 min). Separation-reunion tests evaluated parental pair bond. Egg yolk testosterone and androstenedione levels were evaluated, and the somatic and behavioural development of C and NC chicks was assessed. Our results revealed that members of C pairs were attached to their mates and, although no significant differences in androgen levels could be evidenced between egg sets, a higher proportion of C pairs' eggs were fertilized and their chicks appeared less emotive and more social. Our results revealed that the parental relationship can modulate the behavioural development of their offspring, probably via non-genetic effects, and this could play a major role in the emergence of inter-individual variability.


The occurrence of polyenoic fatty acids with greater than 22 carbon atoms in mammalian spermatozoa.
Fatty acids with carbon chain lengths greater than 22 (VLCFA) have been detected in boar, ram, bull and human spermatozoa. Saturated and mono-unsaturated fatty acids were present in all spermatozoa but, except for human spermatozoa, polyenoic fatty acids were quantitatively the most important components. Marked differences in polyenoic fatty acid composition were observed. Whereas human spermatozoa contain predominantly di-, tri- and tetraenoic fatty acids with up to 32 carbon atoms, boar, ram and bull spermatozoa also contain pentaenoic and/or hexaenoic acids with up to 34 carbon atoms. Human and boar spermatozoa differ markedly from those of the ram and bull in that only n-6 series acids are present.


A multiscale natural community and species-level vulnerability assessment of the Gulf Coast, USA.
Vulnerability assessments combine quantitative and qualitative evaluations of the exposure, sensitivity, and adaptive capacity of species or natural communities to current and future threats. When combined with the economic, ecological or evolutionary value of the species, vulnerability assessments quantify the relative risk to regional species and natural communities and can enable informed prioritization of conservation efforts. Vulnerability assessments are common practice in conservation biology, including the potential impacts of future climate scenarios. However, geographic variation in scenarios and vulnerabilities is rarely quantified. This gap is particularly limiting for informing ecosystem management given that conservation practices typically vary by sociopolitical boundaries rather than by ecological boundaries. To support prioritization of conservation actions across a range of spatial scales, we conducted the Gulf Coast Vulnerability Assessment (GCVA) for four natural communities and eleven focal species around the Gulf of Mexico based on current and future threats from climate change and land-use practices out to 2060. We used the Standardized Index of Vulnerability and Value (SIVVA) tool to assess both natural community and species vulnerabilities. We observed greater variation across ecologically delineated subregions within the Gulf Coast of the U.S. than across climate scenarios. This novel finding suggests that future vulnerability assessments incorporate regional variation and that conservation prioritization may vary across ecological subregions. Across subregions and climate scenarios the most prominent threats were legacy effects, primarily from habitat loss and degradation, that compromised the adaptive capacity of species and natural communities. The second most important threats were future threats from sea-level rise. Our results suggest that the substantial threats species and natural communities face from climate change and sea-level rise would be within their adaptive capacity were it not for historic habitat loss, fragmentation, and degradation.


Crossmodal exogenous orienting improves the accuracy of temporal order judgments.
Although many studies have demonstrated that crossmodal exogenous orienting can lead to a facilitation of reaction times, the issue of whether exogenous spatial orienting also affects the accuracy of perceptual judgments has proved to be much more controversial. Here, we examined whether or not exogenous spatial attentional orienting would affect sensitivity in a temporal discrimination task. Participants judged which of the two target letters, presented on either the same or opposite sides, had been presented first. A spatially non-predictive tone was presented 200 ms prior to the onset of the first visual stimulus. In two experiments, we observed improved performance (i.e., a decrease in the just noticeable difference) when the target letters were presented on opposite sides and the auditory cue was presented on the side of the first visual stimulus, even when central fixation was monitored ("Experiment 2"). A shift in the point of subjective simultaneity was also observed in both experiments, indicating 'prior entry' for cued as compared to uncued first target trials. No such JND or PSS effects were observed when the auditory tone was presented after the second visual stimulus ("Experiment 3"), thus confirming the attentional nature of the effects observed. These findings clearly show that the crossmodal exogenous orienting of spatial attention can affect the accuracy of temporal judgments.


Sequential recruitment in the discrimination of velocity.
Human observers can discriminate a 5% difference in velocity for a wide range of velocities. Using an apparent-motion stimulus, we demonstrated that velocity discrimination depends on the detection of small changes in asynchrony, changes of the order of 1 msec or less. The simplest component of an apparent-motion stimulus is a pair of spatially separate lines presented asynchronously. Generally the incremental asynchrony threshold for a single pair of lines is much too large to account for velocity discrimination. A sequence of five to eight asynchronously presented targets, equivalent to continuous motion viewed for a duration of 80-100 msec is required to reach asymptotic velocity discrimination. Our experiments rule out probability summation as the explanation for the enhanced temporal sensitivity observed with the sequential presentation of multiple asynchronous targets. Sequential recruitment, a descriptive term for this enhanced temporal sensitivity, depends on the summation of a velocity-specific signal within the physiological network responding to motion.


Exploring young people's expectations and experiences of discussing sexual and relationship health with professionals in a children's hospital.
The purpose of this study was to explore young people's expectations and experiences of discussing sexual and relationship health with healthcare professionals whilst attending or having been admitted to an acute children's hospital. Sexual health and teenage pregnancy are topics of debate in the context of public health. Programmes that provide sexual and relationship education to young people are often delivered in education and/or community health settings. However, a significant proportion of young people access acute health services, yet the literature on acute service providers opportunistically providing sexual and relationship advice to young people is limited. Survey. One hundred young people attending either an outpatient appointment or being discharged following an in-patient episode on a hospital ward were approached to complete a questionnaire. Discussion of the questionnaire findings was undertaken with a group of eight young people, which enriched the interpretation and provided clarity of the findings. The topic of sexual and relationship health was important to young people. Although young people infrequently sought advice when in acute settings, they wanted to know there was a choice to discuss these issues with healthcare professionals in the hospital. Young people suggested that professionals needed to have a level of sensitivity and confidence to be able to initiate sexual and relationship discussions with young people. The information from the questionnaires and the discussion group highlighted that young people feel they need to know there is an option to discuss sexual and relationship health whilst attending or being admitted to an acute children's hospital. By giving young people a choice to participate or decline in dialogues about sexual and relationship health, a degree of empowerment and level of respect can be encouraged between young people and healthcare professionals. Healthcare professionals need to be mindful of opportunities to engage young people in maximising their health and well-being, this includes providing them with a choice to discuss sexual and relationship health issues in any health setting.


Hybrid Manganese Dioxide Nanoparticles Potentiate Radiation Therapy by Modulating Tumor Hypoxia.
Hypoxia in the tumor microenvironment (TME) mediates resistance to radiotherapy and contributes to poor prognosis in patients receiving radiotherapy. Here we report the design of clinically suitable formulations of hybrid manganese dioxide (MnO2) nanoparticles (MDNP) using biocompatible materials to reoxygenate the TME by reacting with endogenous H2O2 MDNP containing hydrophilic terpolymer-protein-MnO2 or hydrophobic polymer-lipid-MnO2 provided different oxygen generation rates in the TME relevant to different clinical settings. In highly hypoxic murine or human xenograft breast tumor models, we found that administering either MDNP formulation before radiotherapy modulated tumor hypoxia and increased radiotherapy efficacy, acting to reduce tumor growth, VEGF expression, and vascular density. MDNP treatment also increased apoptosis and DNA double strand breaks, increasing median host survival 3- to 5-fold. Notably, in the murine model, approximately 40% of tumor-bearing mice were tumor-free after a single treatment with MDNPs plus radiotherapy at a 2.5-fold lower dose than required to achieve the same curative treatment without MDNPs. Overall, our findings offer a preclinical proof of concept for the use of MDNP formulations as effective radiotherapy adjuvants. Cancer Res; 76(22); 6643-56. ©2016 AACR.


Array analysis for T-cell associated cytokines in gingival crevicular fluid: Identifying altered profiles associated with periodontal disease status.
Cytokine networks regulate innate and adaptive immune responses, which in turn are recognised to direct the progression or arrest of periodontal disease. This study aimed to compare the profile of seven cytokines, implicated in regulating T-cell networks, in gingival crevicular fluid (GCF) samples with differing classification of periodontal status. GCF samples were collected from patients with strong clinical evidence for chronic periodontitis, aggressive periodontitis, gingivitis or no gingival inflammation. Cytokines IL-6, IFN-ɣ, IL-4, IL-2, IL-17 A, IL10 and TNFα were measured in each sample using a commercial cytometric bead array assay. Descriptive statistics were used to indicate central tendency, data scatter and analysis of variance for each cytokine concentrations between respective patient groups. Heat maps with dendrograms were produced to visualise hierarchical clustering and trends within the data. Median concentrations for all cytokines analysed were highest for gingivitis samples and lowest for aggressive periodontitis samples. The median concentration of IL-6 in gingivitis samples was observed to be 10.5 fold higher (˜17,300 pg/μl) than IL-6 in aggressive periodontitis samples (˜1600 pg/μl). Median concentrations of IL-10, IL-17 A and TNFα were also 2-2.2 fold higher in gingivitis samples compared to aggressive periodontitis. Descriptive statistical analysis noted raised concentrations of IL-6, IL-17 A and TNFα associated with gingivitis; pro-inflammatory cytokines usually associated with periodontal tissue destruction, including bone. Our results would suggest that these cytokines can additionally provide protective roles in preventing progression to advanced forms of periodontal disease. Potential for how these cytokines contribute to providing this role is discussed. Defining the roles for the many cytokines involved in the pathogenesis of periodontal disease is far from complete. Consequently the results of this study serve to evidence proposals that cytokines can exhibit both pro- and anti-inflammatory effects, which is dependent on the signalling environment within which they exist and the antagonizing or modifying actions of other cytokines. Whilst future research is necessary to explore mechanistic action, our study contributes new knowledge suggesting that IL-6 and IL-17 A can provide roles in stabilising the lesion to limit disease progression, which does not preclude alternative roles in promoting periodontal bone loss in advanced forms of disease progression, which is also documented in the literature.


Color-Biased Regions of the Ventral Visual Pathway Lie between Face- and Place-Selective Regions in Humans, as in Macaques.
The existence of color-processing regions in the human ventral visual pathway (VVP) has long been known from patient and imaging studies, but their location in the cortex relative to other regions, their selectivity for color compared with other properties (shape and object category), and their relationship to color-processing regions found in nonhuman primates remain unclear. We addressed these questions by scanning 13 subjects with fMRI while they viewed two versions of movie clips (colored, achromatic) of five different object classes (faces, scenes, bodies, objects, scrambled objects). We identified regions in each subject that were selective for color, faces, places, and object shape, and measured responses within these regions to the 10 conditions in independently acquired data. We report two key findings. First, the three previously reported color-biased regions (located within a band running posterior-anterior along the VVP, present in most of our subjects) were sandwiched between face-selective cortex and place-selective cortex, forming parallel bands of face, color, and place selectivity that tracked the fusiform gyrus/collateral sulcus. Second, the posterior color-biased regions showed little or no selectivity for object shape or for particular stimulus categories and showed no interaction of color preference with stimulus category, suggesting that they code color independently of shape or stimulus category; moreover, the shape-biased lateral occipital region showed no significant color bias. These observations mirror results in macaque inferior temporal cortex (Lafer-Sousa and Conway, 2013), and taken together, these results suggest a homology in which the entire tripartite face/color/place system of primates migrated onto the ventral surface in humans over the course of evolution. Here we report that color-biased cortex is sandwiched between face-selective and place-selective cortex on the bottom surface of the brain in humans. This face/color/place organization mirrors that seen on the lateral surface of the temporal lobe in macaques, suggesting that the entire tripartite system is homologous between species. This result validates the use of macaques as a model for human vision, making possible more powerful investigations into the connectivity, precise neural codes, and development of this part of the brain. In addition, we find substantial segregation of color from shape selectivity in posterior regions, as observed in macaques, indicating a considerable dissociation of the processing of shape and color in both species.


[Efficacy of therapeutic use of ultrasound and sinusoidal modulated currents combed with halotherapy in patient with occupational toxic-dust bronchitis].
Immunological and cardiorespiratory characteristics were studied in 88 alloy industry workers with occupational toxic-dust bronchitis who received the following therapy: sinusoidal modulated currents (SMC), ultrasound (US) on the chest, halotherapy (HT) (52 patients, group 1); SMC + HT (10 patients, group 2); US + HT (15 patients, group 3); HT (11 patients, group 4). The patients did also therapeutic exercise and were massaged (chest). It was found that device physiotherapy (SMC, US) in combination with HT raise the treatment efficacy to 86.5%. This combined treatment is recommended both for treatment and prevention of obstructive syndrome in toxic-dust bronchitis.


Association between diabetes and subsequent Parkinson disease: A record-linkage cohort study.
To investigate the association between type 2 diabetes mellitus (T2DM) and subsequent Parkinson disease (PD). Linked English national Hospital Episode Statistics and mortality data (1999-2011) were used to conduct a retrospective cohort study. A cohort of individuals admitted for hospital care with a coded diagnosis of T2DM was constructed, and compared to a reference cohort. Subsequent PD risk was estimated using Cox regression models. Individuals with a coded diagnosis of cerebrovascular disease, vascular parkinsonism, drug-induced parkinsonism, and normal pressure hydrocephalus were excluded from the analysis. A total of 2,017,115 individuals entered the T2DM cohort and 6,173,208 entered the reference cohort. There were significantly elevated rates of PD following T2DM (hazard ratio [HR] 1.32, 95% confidence interval [CI] 1.29-1.35; p < 0.001). The relative increase was greater in those with complicated T2DM (HR 1.49, 95% CI 1.42-1.56) and when comparing younger individuals (HR 3.81, 95% CI 2.84-5.11 in age group 25-44 years). We report an increased rate of subsequent PD following T2DM in this large cohort study. These findings may reflect shared genetic predisposition and/or disrupted shared pathogenic pathways with potential clinical and therapeutic implications.


Femtosecond laser photodisruption of vitelline vessels of avian embryos as a technique to study embryonic vascular remodeling.
During cardiogenesis, congenital heart defects (CHDs), generally start as local tissue abnormalities without underlying genetic causes, suggesting abnormal hemodynamics may be an important source. Due to the scarcity of experimental techniques that permits the formation of minimally-invasive and well-controlled cardiac perturbations, experimental investigation of embryonic development of CHD via in-vivo models is difficult. In this study, in order to investigate the relationship between abnormal mechanical signaling and embryonic CHD development, a previously developed laser-based technique was adopted to alter chicken embryonic cardiovascular development. The technique incorporates two-photon fluorescence microscopy to visualize deep tissue while femtosecond-pulsed laser photodisruption is used to ablate targeted tissue. Vitelline vessel remodeling under abnormal hemodynamics was the prime concern of the study. In order to alter the hemodynamics, blood flowing inside 50-300 µm diameter Hamburger-Hamilton 24 embryonic vessels was selectively ablated. Red blood cells in the blood and endothelial cells of the vessel walls were damaged as a result of ablation. Cellular injuries led to micro-occlusions in the vessels. Several micro-occlusions formed stable clots, resulting in a complete cessation of blood flow in the targeted vessels. By measuring blood velocities in the surrounding vessels via line scanning technique, the subsequent redistribution of blood flow in the immediate upstream and downstream vessels was revealed. The network was analyzed after 24 h, and it was found to be degraded. Degradation of the entire network can be attributed to the abnormalities in hemodynamics within the vessels. For studying embryonic development of heart defects under disturbed flow conditions, the present study can be extended to clot a blood vessel inside the embryo or a vitelline vessel in the vicinity of the heart. These results demonstrate that, laser-based noninvasive tools should be considered as powerful techniques to analyze hemodynamic signals encountered in embryonic development of CHD.


Characteristics of patients with heart failure with preserved ejection fraction in primary care: a cross-sectional analysis.
Many patients with heart failure with preserved ejection fraction (HFpEF) are undiagnosed, and UK general practice registers do not typically record heart failure (HF) subtype. Improvements in management of HFpEF is dependent on improved identification and characterisation of patients in primary care. To describe a cohort of patients recruited from primary care with suspected HFpEF and compare patients in whom HFpEF was confirmed and refuted. Baseline data from a longitudinal cohort study of patients with suspected HFpEF recruited from primary care in two areas of England. A screening algorithm and review were used to find patients on HF registers without a record of reduced ejection fraction (EF). Baseline evaluation included cardiac, mental and physical function, clinical characteristics, and patient reported outcomes. Confirmation of HFpEF was clinically adjudicated by a cardiologist. In total, 93 (61%) of 152 patients were confirmed HFpEF. The mean age of patients with HFpEF was 79 years, 46% were female, 80% had hypertension, and 37% took ≥10 medications. Patients with HFpEF were more likely to be obese, pre-frail or frail, report more dyspnoea and fatigue, were more functionally impaired, and less active than patients in whom HFpEF was refuted. Few had attended cardiac rehabilitation. Patients with confirmed HFpEF had frequent multimorbidity, functional impairment, frailty, and polypharmacy. Although comorbid conditions were similar between people with and without HFpEF, the former had more obesity, symptoms, and worse physical function. These findings highlight the potential to optimise wellbeing through comorbidity management, medication rationalisation, rehabilitation, and supported self-management.


Ferumoxytol: a new, clinically applicable label for stem-cell tracking in arthritic joints with MRI.
To develop a clinically applicable MRI technique for tracking stem cells in matrix-associated stem-cell implants, using the US FDA-approved iron supplement ferumoxytol. Ferumoxytol-labeling of adipose-derived stem cells (ADSCs) was optimized in vitro. A total of 11 rats with osteochondral defects of both femurs were implanted with ferumoxytol- or ferumoxides-labeled or unlabeled ADSCs, and underwent MRI up to 4 weeks post matrix-associated stem-cell implant. The signal-to-noise ratio of different matrix-associated stem-cell implant was compared with t-tests and correlated with histopathology. An incubation concentration of 500 µg iron/ml ferumoxytol and 10 µg/ml protamine sulfate led to significant cellular iron uptake, T2 signal effects and unimpaired ADSC viability. In vivo, ferumoxytol- and ferumoxides-labeled ADSCs demonstrated significantly lower signal-to-noise ratio values compared with unlabeled controls (p < 0.01). Histopathology confirmed engraftment of labeled ADSCs, with slow dilution of the iron label over time. Ferumoxytol can be used for in vivo tracking of stem cells with MRI.


Glucocorticoids induce rapid up-regulation of mitogen-activated protein kinase phosphatase-1 and dephosphorylation of extracellular signal-regulated kinase and impair proliferation in human and mouse osteoblast cell lines.
A central feature of glucocorticoid (GC)-induced osteoporosis is decreased bone formation, secondary to decreased numbers of functional osteoblasts. We find that ERK activity is essential for serum-induced osteoblast proliferation in vitro because inhibition of MAPK/ERK kinase activity by U0126 completely abolished both serum-induced activation of ERK and proliferation of mouse (MBA-15.4) and human (MG-63) osteoblast cell lines. Dexamethasone (Dex) rapidly (<2 h) inhibits the sustained phase of ERK activation, required for nuclear shift and mitogenesis. This inhibition is reversed by cotreatment with the protein synthesis inhibitor, cycloheximide, and by the GC receptor antagonist, RU486, suggesting a classical transcriptional mechanism. Phosphatase activity was up-regulated by Dex treatment, and inhibition of ERK activity by Dex was also reversed by the protein tyrosine phosphatase inhibitor, vanadate. Coupled with the rapidity of Dex action, this indicates immediate-early gene phosphatase involvement, and we therefore used quantitative, real-time PCR to examine expression profiles of the dual-specificity MAPK phosphatases, MKP-1 and MKP-3. MKP-1, but not MKP-3, mRNA expression was 10-fold up-regulated in both mouse and human osteoblast cell lines within 30 min of Dex treatment and remained elevated for 24 h. MKP-1 protein was also markedly up-regulated following 1-8 h of Dex treatment, and this correlated precisely with dephosphorylation of ERK. Cell proliferation was impaired by Dex treatment, and this was reversed by both RU486 and vanadate. Therefore, MKP-1 up-regulation provides a novel and rapid mechanism, whereby GCs inhibit osteoblast proliferation.


On the length, weight and GC content of the human genome.
Basic parameters commonly used to describe genomes including length, weight and relative guanine-cytosine (GC) content are widely cited in absence of a primary source. By using updated data and original software we determined these values to the best of our knowledge as standard reference for the whole human nuclear genome, for each chromosome and for mitochondrial DNA. We also devised a method to calculate the relative GC content in the whole messenger RNA sequence set and in transcriptomes by multiplying the GC content of each gene by its mean expression level. The male nuclear diploid genome extends for 6.27 Gigabase pairs (Gbp), is 205.00 cm (cm) long and weighs 6.41 picograms (pg). Female values are 6.37 Gbp, 208.23 cm, 6.51 pg. The individual variability and the implication for the DNA informational density in terms of bits/volume were discussed. The genomic GC content is 40.9%. Following analysis in different transcriptomes and species, we showed that the greatest deviation was observed in the pathological condition analysed (trisomy 21 leukaemic cells) and in Caenorhabditis elegans. Our results may represent a solid basis for further investigation on human structural and functional genomics while also providing a framework for other genome comparative analysis.


Complex mammalian-like haematopoietic system found in a colonial chordate.
Haematopoiesis is an essential process that evolved in multicellular animals. At the heart of this process are haematopoietic stem cells (HSCs), which are multipotent and self-renewing, and generate the entire repertoire of blood and immune cells throughout an animal's life1. Although there have been comprehensive studies on self-renewal, differentiation, physiological regulation and niche occupation in vertebrate HSCs, relatively little is known about the evolutionary origin and niches of these cells. Here we describe the haematopoietic system of Botryllus schlosseri, a colonial tunicate that has a vasculature and circulating blood cells, and interesting stem-cell biology and immunity characteristics2-8. Self-recognition between genetically compatible B. schlosseri colonies leads to the formation of natural parabionts with shared circulation, whereas incompatible colonies reject each other3,4,7. Using flow cytometry, whole-transcriptome sequencing of defined cell populations and diverse functional assays, we identify HSCs, progenitors, immune effector cells and an HSC niche, and demonstrate that self-recognition inhibits allospecific cytotoxic reactions. Our results show that HSC and myeloid lineage immune cells emerged in a common ancestor of tunicates and vertebrates, and also suggest that haematopoietic bone marrow and the B. schlosseri endostyle niche evolved from a common origin.


A de novo 2q35-q36.1 deletion incorporating IHH in a Chinese boy (47,XYY) with syndactyly, type III Waardenburg syndrome, and congenital heart disease.
Reports of terminal and interstitial deletions of the long arm of chromosome 2 are rare in the literature. Here, we present a case report concerning a Chinese boy with a 47,XYY karyotype and a de novo deletion comprising approximately 5 Mb between 2q35 and q36.1, along with syndactyly, type III Waardenburg syndrome, and congenital heart disease. High-resolution chromosome analysis to detect copy number variations was carried out using an Affymetrix microarray platform, and the genes affected by the patient's deletion, including IHH, were determined. However, no copy number changes were observed in his healthy parents. The present case exhibited novel syndactyly features, broadening the spectrum of clinical findings observed in individuals with 2q interstitial deletions. Our data, together with previous observations, suggest that IHH haploinsufficiency is the principal pathogenic factor in the syndactyly phenotype in this study, and that different types of variations at the IHH locus may cause divergent disease phenotypes. This is the first report of the involvement of IHH haploinsufficiency in syndactyly phenotype.


Alectinib versus crizotinib in patients with ALK-positive non-small-cell lung cancer (J-ALEX): an open-label, randomised phase 3 trial.
Alectinib, a potent, highly selective, CNS-active inhibitor of anaplastic lymphoma kinase (ALK), showed promising efficacy and tolerability in the single-arm phase 1/2 AF-001JP trial in Japanese patients with ALK-positive non-small-cell lung cancer. Given those promising results, we did a phase 3 trial to directly compare the efficacy and safety of alectinib and crizotinib. J-ALEX was a randomised, open-label, phase 3 trial that recruited ALK inhibitor-naive Japanese patients with ALK-positive non-small-cell lung cancer, who were chemotherapy-naive or had received one previous chemotherapy regimen, from 41 study sites in Japan. Patients were randomly assigned (1:1) via an interactive web response system using a permuted-block method stratified by Eastern Cooperative Oncology Group performance status, treatment line, and disease stage to receive oral alectinib 300 mg twice daily or crizotinib 250 mg twice daily until progressive disease, unacceptable toxicity, death, or withdrawal. The primary endpoint was progression-free survival assessed by an independent review facility. The efficacy analysis was done in the intention-to-treat population, and safety analyses were done in all patients who received at least one dose of the study drug. The study is ongoing and patient recruitment is closed. This study is registered with the Japan Pharmaceutical Information Center (number JapicCTI-132316). Between Nov 18, 2013, and Aug 4, 2015, 207 patients were recruited and assigned to the alectinib (n=103) or crizotinib (n=104) groups. At data cutoff for the second interim analysis, 24 patients in the alectinib group had discontinued treatment compared with 61 in the crizotinib group, mostly due to lack of efficacy or adverse events. At the second interim analysis (data cutoff date Dec 3, 2015), an independent data monitoring committee determined that the primary endpoint of the study had been met (hazard ratio 0·34 [99·7% CI 0·17-0·71], stratified log-rank p<0·0001) and recommended an immediate release of the data. Median progression-free survival had not yet been reached with alectinib (95% CI 20·3-not estimated) and was 10·2 months (8·2-12·0) with crizotinib. Grade 3 or 4 adverse events occurred at a greater frequency with crizotinib (54 [52%] of 104) than alectinib (27 [26%] of 103). Dose interruptions due to adverse events were also more prevalent with crizotinib (77 [74%] of 104) than with alectinib (30 [29%] of 103), and more patients receiving crizotinib (21 [20%]) than alectinib (nine [9%]) discontinued the study drug because of an adverse event. No adverse events with a fatal outcome occurred in either treatment group. These results provide the first head-to-head comparison of alectinib and crizotinib and have the potential to change the standard of care for the first-line treatment of ALK-positive non-small-cell lung cancer. The dose of alectinib (300 mg twice daily) used in this study is lower than the approved dose in countries other than Japan; however, this limitation is being addressed in the ongoing ALEX study. Chugai Pharmaceutical Co, Ltd.


Contact Map Fingerprints of Protein-Ligand Unbinding Trajectories Reveal Mechanisms Determining Residence Times Computed from Scaled Molecular Dynamics.
The binding kinetic properties of potential drugs may significantly influence their subsequent clinical efficacy. Predictions of these properties based on computer simulations provide a useful alternative to their expensive and time-consuming experimental counterparts, even at an early drug discovery stage. Herein, we perform scaled molecular dynamics (ScaledMD) simulations on a set of 27 ligands of HSP90 belonging to more than seven chemical series to estimate their relative residence times. We introduce two new techniques for the analysis and the classification of the simulated unbinding trajectories. The first technique, which helps in estimating the limits of the free energy well around the bound state, and the second one, based on a new contact map fingerprint, allow the description and the comparison of the paths that lead to unbinding. Using these analyses, we find that ScaledMD's relative residence time generally enables the identification of the slowest unbinders. We propose an explanation for the underestimation of the residence times of a subset of compounds, and we investigate how the biasing in ScaledMD can affect the mechanistic insights that can be gained from the simulations.


Experience of palliative home care according to caregivers' and patients' ages in Hong Kong Chinese people.
To identify the relationship between family caregivers' reported difficulty in managing caregiver tasks and ages of caregivers and patients. Cross-sectional, descriptive survey. A hospice homecare program in Hong Kong. Twenty-nine Chinese family caregivers who had experienced at least weekly caregiving responsibility for more than two months and were able to read and understand Chinese. Respondents completed a caregiver task inventory. Four homecare nurses assisted in the distribution and collection of questionnaires. Caregiver tasks and age and patient age. Caregivers' age was negatively correlated with reported difficulty in overall tasks and in interpersonal ties. The patients' age was negatively correlated with reported difficulty in direct care to patients, intrapersonal tasks, and overall tasks. The younger the caregiver, the more difficulty he or she experienced in the caregiving role, particularly in the maintenance of social and family ties. Caregivers of younger patients experienced more difficulty in most aspects of caregiving tasks. More research with a larger sample size is required to fully investigate the effect of age on the family caregiving experience and the validity of the caregiver task inventory. Nursing support and preparation to younger caregivers and caregivers of younger patients are suggested in the practice of palliative home care.


Evidence of multiple Treponema phylotypes involved in bovine digital dermatitis as shown by 16S rRNA gene analysis and fluorescence in situ hybridization.
The etiopathogenesis of the skin disease digital dermatitis (DD), an important cause of lameness in cattle, remains uncertain. Microscopically, the disease appears to be polymicrobial, with spirochetes as the predominant bacteria. The objective of this study was to identify the main part of the bacteria involved in DD lesions of cattle by using culture-independent molecular methods. Ten different phylotypes of Treponema were identified either by 16S rRNA gene sequencing of bacteria from DD lesions or by fluorescence in situ hybridization (FISH) analysis using phylotype-specific 16S rRNA-directed oligonucleotide probes. Two phylotypes, phylotype 1 (PT1) and PT2, were not closely related to any characterized treponemal species. PT7 was 99.3% identical to Treponema denticola, while PT9 resembled T. vincentii by 96%. The remaining phylotypes, PT3, PT4, PT5, PT6, and PT8, and Treponema brennaborense had previously been isolated from DD lesions. Forty DD biopsy specimens were examined for Treponema by FISH. With one exception, all of the biopsy specimens revealed epidermotropic, intermingled infection with three or more different phylotypes (mean, 4.7). The most prevalent species were PT1 (95%), PT6 (93%), and PT3 (85%). While colonization by PT3 was confined to the surface of the epidermis, both PT1 and PT6 invaded deep into the stratum spinosum and were seen in ulcerated dermal papillae. In two cases, all 10 phylotypes were demonstrated. Furthermore, FISH with a Treponema group-specific probe showed that Treponema accounted for more than 90% of the total bacterial population in the biopsy specimens. These data strongly suggest that a group of apparently symbiotic Treponema species are involved as primary bacterial pathogens in DD.


Increasing security through public health: a practical model.
As political and social changes sweep the globe, there are opportunities to increase national security through innovative approaches. While traditional security methods such as defense forces and homeland security provide both pre-emptive and defensive protection, new methods could meet emerging challenges by responding to the political, financial, and social trends. One method is the integration of defense, medicine and public health. By assisting a nation by providing basic services, such as healthcare, collaborative efforts can increase stabilization in areas of unrest. Improved health outcomes leads to increased domestic security, which can create a ripple effect across a region. Assessment, uptake and sustainability by the host nation are critical for program success. The proposed methodology focuses on the use of primarily extant resources, such as programs used by Special Operations Forces and other health and defense programs. Additional components include evaluation, set objectives and mission collaborations. As the nexus between foreign affairs, security, and public health is increasingly validated through research and practice, standardized interventions should be developed to minimize overlapping expenditures, promote security and strengthen international relations.


Sofosbuvir: A novel treatment option for chronic hepatitis C infection.
Hepatitis C currently infects more than 170 million people around the world, leading to significant morbidity and mortality. The current standard of care for HCV infection, including one of the two protease inhibitors, telaprevir or boceprevir, for 12-32 weeks, along with pegylated interferon alfa-2a (PEG-IFN-α) and ribavirin for up to 48 weeks, is unsatisfactory in many cases, either because of lack of efficacy or because of treatment-related adverse effects. There is an urgent need of new drugs with improved efficacy as well as a safety profile. Sofosbuvir, a recently approved nucleotide analog, is a highly potent inhibitor of the NS5B polymerase in the Hepatitis C virus (HCV), and has shown high efficacy in combination with several other drugs, with and without PEG-INF, against HCV. It offers many advantages due to its high potency, low side effects, oral administration, and high barrier to resistance. The efficacy and safety were demonstrated in many large and well-designed phase 2 and phase 3 clinical trials like NEUTRINO, PROTON, ELECTRON, ATOMIC, COSMOS, FUSION, FISSION, NUCLEAR, POSITRON, and the like. It is generally well-tolerated. Adverse events that occurred include: Headache, insomnia, fatigue, nausea, dizziness, pruritis, upper respiratory tract infections, rash, back pain, grade 1 anemia, and grade 4 lymphopenia; however, the exact safety profile can only be judged when this drug is actually used on a large scale.


Effect of the use of carprofen in dogs undergoing intense rehabilitation after lateral fabellar suture stabilization.
To determine whether carprofen, a commercially available NSAID, would decrease perceived exertion and signs of pain in dogs and therefore increase muscle mass and hind limb function without decreasing range of motion after lateral fabellar suture stabilization. Randomized, blinded, controlled clinical trial. 35 dogs with cranial cruciate ligament rupture and lateral fabellar suture stabilization followed by rehabilitation. All dogs underwent surgical stabilization of cranial cruciate ligament rupture by placement of a lateral fabellar suture. Dogs received carprofen (2.2 mg/kg [1 mg/lb], PO, q 12 h) for the first 7 days after surgery and underwent concentrated rehabilitation exercises during weeks 3, 5, and 7 after surgery. Eighteen dogs also received carprofen (2.2 mg/kg, PO, q 12 h) during the weeks of concentrated rehabilitation. Outcomes were measured by a single investigator, who was blinded to group assignments, using pressure platform gait analysis, goniometry, thigh circumference, and mean workout speed at a consistent level of exertion. There were no differences between the 2 groups in ground reaction forces, thigh circumference, or exertion (mean workout speed) over time or at any individual time point. However, both groups improved significantly over time for all outcome measures. Providing carprofen to dogs during concentrated rehabilitation after lateral fabellar suture stabilization did not improve hind limb function, range of motion, or thigh circumference, nor did it decrease perceived exertion, compared with control dogs. Carprofen was not a compulsory component of a physical therapy regimen after lateral fabellar suture stabilization.


Registration, documentation, and auditing of stillbirths and neonatal deaths in Jordan from healthcare professionals' perspectives: reality, challenges and suggestions.
Objectives: The current study aimed to explore healthcare professionals' (HCPs) perceptions towards the registration and reporting process of stillbirth and neonatal mortality and their causes. Another aim is to explore HCPs' perceived importance of registering stillbirths and neonatal deaths as well as the perceived challenges and suggestions of building a new surveillance and auditing system to report stillbirths and neonatal deaths in Jordan.Methods: A descriptive qualitative approach using an in-depth focus group discussion was used. A total of 16 focus groups were conducted in four major representative hospitals that cover different geographical areas in Jordan. An average of five healthcare providers (HCPs) was interviewed in each focus group with a total of 80 HCPs. All recorded focus group interviews were transcribed in a full verbatim, which was checked for accuracy by the project team. The whole content thematic analysis process was conducted in its original Arabic language to conserve credibility of the findings. Healthcare providers' perceptions were derived from the group discussions using a group-interview guide.Results: Overall, the majority of HCPs across all hospitals agreed that it is important to document neonatal deaths and stillbirths in the hospital records mainly for statistical purposes. HCPs usually document neonatal deaths but not stillbirths. The causes of stillbirths and neonatal deaths are inaccurately recorded and not usually completed by the attending physician. Surprisingly, only neonates who live more than 4 h after delivery are registered as neonatal deaths but any neonate who dies before 4 h after delivery is not registered or registered as stillbirth. The majority of HCPs said that they were not aware of having neonatal death review committee in their hospitals. Importantly, the majority of HCPs in the four hospitals were enthusiastic about the development of a new surveillance system to register neonatal deaths and stillbirths in Jordan. Several suggestions were conveyed by the HCPs to better build, develop, implement, and sustain the proposed surveillance system.Conclusions: Electronic health information system and centralized database for compiling audit, registering births and deaths, and assigning causes of deaths should be developed and implemented. Designing and implementing an electronic registry or surveillance system that adopts ICD-10 codes is expected not only improve the completeness and timeliness of registration but also results in accurate recording of the causes of deaths.


Sensing picornaviruses using molecular imprinting techniques on a quartz crystal microbalance.
Molecular imprinting techniques were adapted to design a sensor for the human rhinovirus (HRV) and the foot-and-mouth disease virus (FMDV), which are two representatives of picornaviruses. Stamp imprinting procedures lead to patterned polyurethane layers that depict the geometrical features of the template virus, as confirmed by AFM for HRV. Quartz crystal microbalance (QCM) measurements show that the resulting layers incorporate the template viruses reversibly and lead to mass effects that are almost an order of magnitude higher than those of nonspecific adsorption. Thus, for example, the sensor yields a net frequency effect of -300 Hz when applying a virus suspension with a concentration of approximately 100 microg/mL with an excellent signal-to-noise ratio. The cavities are not only selective to shape but also to surface chemistry: different HRV serotypes (HRV1A, HRV2, HRV14, and HRV16, respectively) can be distinguished with the sensor materials by a selectivity factor of 3, regardless of the group (major/minor) to which they belong. The same selectivity factor can be observed between HRV and FMDV. Hence, imprinting leads to an "artificial antibody" toward viruses, which does not only recognize their receptor binding sites, but rather detects the whole virus as an entity. Brunauer-Emmett-Teller (BET) studies allow simulation of the sensor characteristics and reveal the number of favorable binding sites in the coatings.


Ultrafine particle exposure for bicycle commutes in rush and non-rush hour traffic: A repeated measures study in Copenhagen, Denmark.
Ultrafine particles (UFP), harmful to human health, are emitted at high levels from motorized traffic. Bicycle commuting is increasingly encouraged to reduce traffic emissions and increase physical activity, but higher breathing rates increase inhaled UFP concentrations while in traffic. We assessed exposure to UFP while cycling along a fixed 8.5 km inner-city route in Copenhagen, on weekdays over six weeks (from September to October 2020), during morning and afternoon rush-hour, as well as morning non-rush-hour, traffic time periods starting from 07:45, 15:45, and 09:45 h, respectively. Continuous measurements were made (each second) of particle number concentration (PNC) and location. PNC levels were summarized and compared across time periods. We used generalized additive models to adjust for meteorological factors, weekdays and trends. A total of 61 laps were completed, during 28 days (∼20 per time period). Overall mean PNC was 18,149 pt/cm3 (range 256-999,560 pt/cm3) with no significant difference between morning rush-hour (18003 pt/cm3), afternoon rush-hour (17560 pt/cm3) and late morning commute (17560 pt/cm3) [p = 0.85]. There was substantial spatial variation of UFP exposure along the route with highest PNC levels measured at traffic intersections (∼38,000-42000 pt/cm3), multiple lane roads (∼38,000-40000 pt/cm3) and construction sites (∼44,000-51000 pt/cm3), while lowest levels were measured at smaller streets, areas with open built environment (∼12,000 pt/cm3), as well as at a bus-only zone (∼15,000 pt/cm3). UFP exposure in inner-city Copenhagen did not differ substantially when bicycling in either rush-hour or non-rush-hour, or morning or afternoon, traffic time periods. UFP exposure varied substantially spatially, with highest concentrations around intersections, multiple lane roads, and construction sites. This suggests that exposure to UFP is not necessarily reduced by avoiding rush-hours, but by avoiding sources of pollution along the bicycling route.


Changes in markers of oxidative stress and DNA damage in human visceral adipose tissue from subjects with obesity and type 2 diabetes.
In the past 30 years, prevalence of obesity has almost trebled resulting in an increased incidence of type 2 diabetes mellitus and other co-morbidities. Visceral adipose tissue is believed to play a vital role, but underlying mechanisms remain unclear. Our aim was to investigate changes in markers of oxidative damage in human visceral adipose tissue to determine levels of oxidative burden that may be attributed to obesity and/or diabetes. Visceral adipose tissue samples from 61 subjects undergoing abdominal surgery grouped as lean, obese and obese with type 2 diabetes mellitus, were examined using 3 different markers of oxidative stress. Malondialdehyde (MDA) concentration was measured as a marker of lipid peroxidation, telomere length and Comet assay as markers of oxidative DNA damage. No significant difference in MDA concentration, telomere length and DNA damage was observed between groups, although longer telomere lengths were seen in the obese with diabetes group compared to the obese group (P<0.05). Lower MDA concentration and longer telomere length were seen in subjects with diabetes compared to those without (P<0.05). DNA damage, analysed via Comet assay, was significantly lower in subjects with diabetes compared to those without (P<0.05). A paradoxical decrease in oxidative stress and DNA damage was observed in samples from subjects with type 2 diabetes mellitus. Further work is required to investigate this further, however this phenomenon may be due to an up regulation of antioxidant defences in adipose tissue.


Inhibitory effect of alpha-1 adrenoceptor stimulation on cardiac sympathetic neurotransmission in pithed normotensive rats.
In the present study we investigated the inhibitory effect of the selective alpha-1 adrenoceptor agonists cirazoline, amidephrine and St 587 on the cardiac sympathetic neurotransmission in pithed normotensive rats. Increases in heart rate were elicited by electrical stimulation of the cardiac sympathetic nerves or by i.v. administration of norepinephrine, isoproterenol or tyramine. Intravenous pretreatment of the animals with cirazoline, amidephrine or St 587 diminished the heart rate response to sympathetic stimulation significantly. However, the tachycardia produced by norepinephrine, isoproterenol or tyramine was also inhibited significantly by the selective alpha-1 adrenoceptor agonists. The selective alpha-1 antagonist prazosin blocked the sympathoinhibitory effect to alpha-1 adrenoceptor stimulation significantly. However, the inhibitory effect of cirazoline and St 587 was not suppressed completely by a maximally effective dose of prazosin. In contrast, the sympathoinhibitory action of amidephrine was antagonized completely by prazosin. However, the selective alpha-2 antagonist rauwolscine also produced a significant, albeit modest, attenuation of the sympathoinhibitory effect to amidephrine. The results of the present study indicate that alpha-1 adrenoceptor agonists, at relatively high doses, inhibit the sympathetic neurotransmission in rat heart. This sympathoinhibitory effect is mediated largely by alpha-1 adrenoceptors which are localized postjunctionally rather than prejunctionally.


Glibenclamide blockade of CFTR chloride channels.
The cystic fibrosis transmembrane conductance regulator (CFTR) is a protein kinase A- and ATP-regulated Cl- channel located in the apical membranes of epithelial cells. Previously Sheppard and Welsh (J. Gen. Physiol. 100: 573-591, 1992) showed that glibenclamide, a compound which binds to the sulfonylurea receptor and thus blocks nucleotide-dependent K+ channels, reduced CFTR whole cell current. The aim of this study was to identify the mechanism underlying this inhibition in cell-free membrane patches containing CFTR Cl- channels. Exposure to gliben-clamide caused a reversible reduction in current carried by CFTR which was paralleled by a decrease in channel open probability (Po). The decrease in Po was concentration dependent, and half-maximum inhibition (ki) occurred at 30 microM. Fluctuation analysis indicated a flickery-type block of open CFTR channels. Event duration analysis supported this notion by showing that the glibenclamide-induced decrease in Po was accompanied by interruptions of open bursts [i.e., an apparent reduction in the burst duration (Tburst)] with only a slight reduction in closed time (Tc). The plot of the corresponding open-to-closed (Tburst-1) and closed-to-open (Tc-1) rates as a function of glibenclamide concentration were consistent with a pseudo-first-order open-blocked mechanism and provided estimates of the on rate (kon = 1.17 microM-1S-1), the off rate (koff = 16 s-1), and the dissociation constant (Kd = 14 microM). The difference between the Ki (30 microM) and the Kd (14 microM) is the result expected for a closed-open-blocked model with an initial Po of 0.47. Since the initial Po was 0.50 +/- 0.02 (n = 12), we can conclude that glibenclamide blocks CFTR by a closed-open-blocked mechanism.


Adjacency and Area Explain Species Bioregional Shifts in Neotropical Palms.
Environmental and geographical variables are known drivers of community assembly, however their influence on phylogenetic structure and phylogenetic beta diversity of lineages within different bioregions is not well-understood. Using Neotropical palms as a model, we investigate how environmental and geographical variables affect the assembly of lineages into bioregions across an evolutionary time scale. We also determine lineage shifts between tropical (TRF) and non-tropical (non-TRF) forests. Our results identify that distance and area explain phylogenetic dissimilarity among bioregions. Lineages in smaller bioregions are a subset of larger bioregions and contribute significantly to the nestedness component of phylogenetic dissimilarity, here interpreted as evidence for a bioregional shift. We found a significant tendency of habitat shifts occurring preferentially between TRF and non-TRF bioregions (31 shifts) than from non-TRF to TRF (24) or from TRF to TRF (11) and non-TRF to non-TRF (9). Our results also present cases where low dissimilarity is found between TRF and non-TRF bioregions. Most bioregions showed phylogenetic clustering and larger bioregions tended to be more clustered than smaller ones, with a higher species turnover component of phylogenetic dissimilarity. However, phylogenetic structure did not differ between TRF and non-TRF bioregions and diversification rates were higher in only two lineages, Attaleinae and Bactridinae, which are widespread and overabundant in both TRF and non-TRF bioregions. Area and distance significantly affected Neotropical palm community assembly and contributed more than environmental variables. Despite palms being emblematic humid forest elements, we found multiple shifts from humid to dry bioregions, showing that palms are also important components of these environments.


In vitro selection of a DNA-templated small-molecule library reveals a class of macrocyclic kinase inhibitors.
DNA-templated organic synthesis enables the translation of DNA sequences into synthetic small-molecule libraries suitable for in vitro selection. Previously, we described the DNA-templated multistep synthesis of a 13,824-membered small-molecule macrocycle library. Here, we report the discovery of small molecules that modulate the activity of kinase enzymes through the in vitro selection of this DNA-templated small-molecule macrocycle library against 36 biomedically relevant protein targets. DNA encoding selection survivors was amplified by PCR and identified by ultra-high-throughput DNA sequencing. Macrocycles corresponding to DNA sequences enriched upon selection against several protein kinases were synthesized on a multimilligram scale. In vitro assays revealed that these macrocycles inhibit (or activate) the kinases against which they were selected with IC(50) values as low as 680 nM. We characterized in depth a family of macrocycles enriched upon selection against Src kinase, and showed that inhibition was highly dependent on the identity of macrocycle building blocks as well as on backbone conformation. Two macrocycles in this family exhibited unusually strong Src inhibition selectivity even among kinases closely related to Src. One macrocycle was found to activate, rather than inhibit, its target kinase, VEGFR2. Taken together, these results establish the use of DNA-templated synthesis and in vitro selection to discover small molecules that modulate enzyme activities, and also reveal a new scaffold for selective ATP-competitive kinase inhibition.


Nyamanini and midway viruses define a novel taxon of RNA viruses in the order Mononegavirales.
Here, we report the sequencing and classification of Nyamanini virus (NYMV) and Midway virus (MIDWV), two antigenically related viruses that were first isolated in 1957 and 1966, respectively. Although these viruses have been cultured multiple times from cattle egrets, seabirds, and their ticks, efforts to classify them taxonomically using conventional serological and electron microscopic approaches have failed completely. We used a random shotgun sequencing strategy to define the genomes of NYMV and MIDWV. Contigs of 11,631 and 11,752 nucleotides, representing the complete genome of NYMV and the near-complete genome of MIDWV, respectively, were assembled. Each virus genome was predicted to carry six open reading frames (ORFs). BLAST analysis indicated that only two of the ORF proteins of each virus, the putative nucleocapsid and polymerase, had detectable sequence similarity to known viral proteins. Phylogenetic analysis of these ORF proteins demonstrated that the closest relatives of NYNV and MIDWV are negative-stranded-RNA viruses in the order Mononegavirales. On the basis of their very limited sequence similarity to known viruses, we propose that NYMV and MIDWV define a novel genus, Nyavirus, in this order.


Studies toward the Development of Antiproliferative Neoclerodanes from Salvinorin A.
The success rate for central nervous system (CNS) drug candidates in the clinic is relatively low compared to the industry average across other therapeutic areas. Penetration through the blood-brain barrier (BBB) to reach the therapeutic target is a major obstacle in development. The rapid CNS penetration of salvinorin A has suggested that the neoclerodane nucleus offers an excellent scaffold for developing antiproliferative compounds that enter the CNS. The Liebeskind-Srogl reaction was used as the main carbon-carbon bond-forming step toward the synthesis of quinone-containing salvinorin A analogues. Quinone-containing salvinorin A analogues were shown to have antiproliferative activity against the MCF7 breast cancer cell line, but show no significant activity at the κ-opioid receptors. In an in vitro model of BBB penetration, quinone-containing salvinorin A analogues were shown to passively diffuse across the cell monolayer. The analogues, however, are substrates of P-glycoprotein, and thus further modification of the molecules is needed to reduce the affinity for the efflux transporter.


Vesicouterine fistula presenting with urinary incontinence 30 years after primary Cesarean: Case report and review of the literature.
A vesicouterine fistula is an abnormal communication between the bladder and the uterus. The most common cause is lower segment Cesarean section. Conservative treatment may be applied in some cases, but surgery is the definitive treatment. We present a 55-year-old woman with vesicouterine fistula after primary Cesarean section; she presented with urinary incontinence 30 years later. Methylene blue dye test was performed and drainage of urine dyed with methylene blue through the uterine cervix was observed. Diagnosis was confirmed with magnetic resonance imaging. Total abdominal hysterectomy and bilateral oophorectomy was performed and the bladder wall was sutured in a double layer with 2/0 polyglycolic suture. No complications developed and the patient did not suffer from any urinary incontinence afterwards. Vesicouterine fistula should be suspected in cases presenting with urinary incontinence even years after Cesarean section; diagnostic tests and, if necessary, appropriate surgery should be performed.


Electrochemical capacitors: mechanism, materials, systems, characterization and applications.
Electrochemical capacitors (i.e. supercapacitors) include electrochemical double-layer capacitors that depend on the charge storage of ion adsorption and pseudo-capacitors that are based on charge storage involving fast surface redox reactions. The energy storage capacities of supercapacitors are several orders of magnitude higher than those of conventional dielectric capacitors, but are much lower than those of secondary batteries. They typically have high power density, long cyclic stability and high safety, and thus can be considered as an alternative or complement to rechargeable batteries in applications that require high power delivery or fast energy harvesting. This article reviews the latest progress in supercapacitors in charge storage mechanisms, electrode materials, electrolyte materials, systems, characterization methods, and applications. In particular, the newly developed charge storage mechanism for intercalative pseudocapacitive behaviour, which bridges the gap between battery behaviour and conventional pseudocapacitive behaviour, is also clarified for comparison. Finally, the prospects and challenges associated with supercapacitors in practical applications are also discussed.


Agricultural nematology in East and Southern Africa: problems, management strategies and stakeholder linkages.
By 2050, Africa's population is projected to exceed 2 billion. Africa will have to increase food production more than 50% in the coming 50 years to meet the nutritional requirements of its growing population. Nowhere is the need to increase agricultural productivity more pertinent than in much of Sub-Saharan Africa, where it is currently static or declining. Optimal pest management will be essential, because intensification of any system creates heightened selection pressures for pests. Plant-parasitic nematodes and their damage potential are intertwined with intensified systems and can be an indicator of unsustainable practices. As soil pests, nematodes are commonly overlooked or misdiagnosed, particularly where appropriate expertise and knowledge transfer systems are meager or inadequately funded. Nematode damage to roots results in less efficient root systems that are less able to access nutrients and water, which can produce symptoms typical of water or nutrient deficiency, leading to misdiagnosis of the underlying cause. Damage in subsistence agriculture is exacerbated by growing crops on degraded soils and in areas of low water retention where strong root growth is vital. This review focuses on the current knowledge of economically important nematode pests affecting key crops, nematode control methods and the research and development needs for sustainable management, stakeholder involvement and capacity building in the context of crop security in East and Southern Africa, especially Kenya, Tanzania, Uganda and Zimbabwe.


Intra-tumoral heterogeneity of gemcitabine delivery and mass transport in human pancreatic cancer.
There is substantial heterogeneity in the clinical behavior of pancreatic cancer and in its response to therapy. Some of this variation may be due to differences in delivery of cytotoxic therapies between patients and within individual tumors. Indeed, in 12 patients with resectable pancreatic cancer, we previously demonstrated wide inter-patient variability in the delivery of gemcitabine as well as in the mass transport properties of tumors as measured by computed tomography (CT) scans. However, the variability of drug delivery and transport properties within pancreatic tumors is currently unknown. Here, we analyzed regional measurements of gemcitabine DNA incorporation in the tumors of the same 12 patients to understand the degree of intra-tumoral heterogeneity of drug delivery. We also developed a volumetric segmentation approach to measure mass transport properties from the CT scans of these patients and tested inter-observer agreement with this new methodology. Our results demonstrate significant heterogeneity of gemcitabine delivery within individual pancreatic tumors and across the patient cohort, with gemcitabine DNA incorporation in the inner portion of the tumors ranging from 38 to 74% of the total. Similarly, the CT-derived mass transport properties of the tumors had a high degree of heterogeneity, ranging from minimal difference to almost 200% difference between inner and outer portions of the tumor. Our quantitative method to derive transport properties from CT scans demonstrated less than 5% difference in gemcitabine prediction at the average CT-derived transport value across observers. These data illustrate significant inter-patient and intra-tumoral heterogeneity in the delivery of gemcitabine, and highlight how this variability can be reproducibly accounted for using principles of mass transport. With further validation as a biophysical marker, transport properties of tumors may be useful in patient selection for therapy and prediction of therapeutic outcome.


Cortical transcriptome analysis after spinal cord injury reveals the regenerative mechanism of central nervous system in CRMP2 knock-in mice.
Recent studies have shown that mutation at Ser522 causes inhibition of collapsin response mediator protein 2 (CRMP2) phosphorylation and induces axon elongation and partial recovery of the lost sensorimotor function after spinal cord injury (SCI). We aimed to reveal the intracellular mechanism in axotomized neurons in the CRMP2 knock-in (CRMP2KI) mouse model by performing transcriptome analysis in mouse sensorimotor cortex using micro-dissection punching system. Prior to that, we analyzed the structural pathophysiology in axotomized or neighboring neurons after SCI and found that somatic atrophy and dendritic spine reduction in sensorimotor cortex were suppressed in CRMP2KI mice. Further analysis of the transcriptome has aided in the identification of four hemoglobin genes Hba-a1, Hba-a2, Hbb-bs, and Hbb-bt that are significantly upregulated in wild-type mice with concomitant upregulation of genes involved in the oxidative phosphorylation and ribosomal pathways after SCI. However, we observed substantial upregulation in channel activity genes and downregulation of genes regulating vesicles, synaptic function, glial cell differentiation in CRMP2KI mice. Moreover, the transcriptome profile of CRMP2KI mice has been discussed wherein energy metabolism and neuronal pathways were found to be differentially regulated. Our results showed that CRMP2KI mice displayed improved SCI pathophysiology not only via microtubule stabilization in neurons, but also possibly via the whole metabolic system in the central nervous system, response changes in glial cells, and synapses. Taken together, we reveal new insights on SCI pathophysiology and the regenerative mechanism of central nervous system by the inhibition of CRMP2 phosphorylation at Ser522. All these experiments were performed in accordance with the guidelines of the Institutional Animal Care and Use Committee at Waseda University, Japan (2017-A027 approved on March 21, 2017; 2018-A003 approved on March 25, 2018; 2019-A026 approved on March 25, 2019).


Isolation and identification of ingredients inducing cancer cell death from the seeds of Alpinia galanga, a Chinese spice.
This study was carried out to isolate ingredients from the seeds of a Chinese spice (Alpinia galangal) and to evaluate their cytotoxic activity on cancer cell lines. Isolation and purification of the phytochemical constituents were conducted using silica gel, Sephadex LH-20 and ODS columns. After extraction using 95% ethanol, the total extracts were re-extracted, resulting in petroleum ether (PE), ethyl acetate (EA) and water fractions, respectively. Activity tests showed that the EA fraction exhibited obvious (p < 0.05) protective effects on H2O2 damaged PC-12 cells at 20 μg mL(-1), and showed much higher (p < 0.05) cytotoxic activity on cancer cell lines than other fractions. Five compounds, 1'-S-1'-acetoxyeugenol acetate (I), 1'-S-1'-acetoxychavicol acetate (II), 2-propenal, 3-[4-(acetyloxy)-3-methoxyphenyl] (III), isocoronarin D (IV) and caryolane-1, 9β-diol (V), were obtained from the EA fraction and identified by HPLC, UV, MS, and NMR spectroscopic analyses. Compounds III and V were isolated from A. galangal for the first time. Moreover, compounds I, II, IV and V were the main active ingredients for inducing death of the tested cancer cells, and their IC50 values ranged from 60 to 90 μg mL(-1), indicating that these compounds possessed a wide anti-cancer capability. Therefore, A. galangal seeds could be a potential source of healthy food for tumor prevention.


Genetic interactions among floral homeotic genes of Arabidopsis.
We describe allelic series for three loci, mutations in which result in homeotic conversions in two adjacent whorls in the Arabidopsis thaliana flower. Both the structure of the mature flower and its development from the initial primordium are described by scanning electron microscopy. New mutations at the APETALA2 locus, ap2-2, ap2-8 and ap2-9, cause homeotic conversions in the outer two whorls: sepals to carpels (or leaves) and petals to stamens. Two new mutations of PISTILLATA, pi-2 and pi-3, cause second and third whorl organs to differentiate incorrectly. Homeotic conversions are petals to sepals and stamens to carpels, a pattern similar to that previously described for the apetala3-1 mutation. The AGAMOUS mutations, ag-2 and ag-3, affect the third and fourth whorls and cause petals to develop instead of stamens and another flower to arise in place of the gynoecium. In addition to homeotic changes, mutations at the APETALA2, APETALA3 and PISTILLATA loci may lead to reduced numbers of organs, or even their absence, in specific whorls. The bud and flower phenotypes of doubly and triply mutant strains, constructed with these and previously described alleles, are also described. Based on these results, a model is proposed that suggests that the products of these homeotic genes are each active in fields occupying two adjacent whorls, AP2 in the two outer whorls, PI and AP3 in whorls two and three, and AG in the two inner whorls. In combination, therefore, the gene products in these three concentric, overlapping fields specify the four types of organs in the wild-type flower. Further, the phenotypes of multiple mutant lines indicate that the wild-type products of the AGAMOUS and APETALA2 genes interact antagonistically. AP2 seems to keep the AG gene inactive in the two outer whorls while the converse is likely in the two inner whorls. This field model successfully predicts the phenotypes of all the singly, doubly and triply mutant flowers described.


Evaluation and Referral for Developmental Dysplasia of the Hip in Infants.
Developmental dysplasia of the hip (DDH) encompasses a wide spectrum of clinical severity, from mild developmental abnormalities to frank dislocation. Clinical hip instability occurs in 1% to 2% of full-term infants, and up to 15% have hip instability or hip immaturity detectable by imaging studies. Hip dysplasia is the most common cause of hip arthritis in women younger than 40 years and accounts for 5% to 10% of all total hip replacements in the United States. Newborn and periodic screening have been practiced for decades, because DDH is clinically silent during the first year of life, can be treated more effectively if detected early, and can have severe consequences if left untreated. However, screening programs and techniques are not uniform, and there is little evidence-based literature to support current practice, leading to controversy. Recent literature shows that many mild forms of DDH resolve without treatment, and there is a lack of agreement on ultrasonographic diagnostic criteria for DDH as a disease versus developmental variations. The American Academy of Pediatrics has not published any policy statements on DDH since its 2000 clinical practice guideline and accompanying technical report. Developments since then include a controversial US Preventive Services Task Force "inconclusive" determination regarding usefulness of DDH screening, several prospective studies supporting observation over treatment of minor ultrasonographic hip variations, and a recent evidence-based clinical practice guideline from the American Academy of Orthopaedic Surgeons on the detection and management of DDH in infants 0 to 6 months of age. The purpose of this clinical report was to provide literature-based updated direction for the clinician in screening and referral for DDH, with the primary goal of preventing and/or detecting a dislocated hip by 6 to 12 months of age in an otherwise healthy child, understanding that no screening program has eliminated late development or presentation of a dislocated hip and that the diagnosis and treatment of milder forms of hip dysplasia remain controversial.


Micro-Level Adaptation, Macro-Level Selection, and the Dynamics of Market Partitioning.
This paper provides a micro-foundation for dual market structure formation through partitioning processes in marketplaces by developing a computational model of interacting economic agents. We propose an agent-based modeling approach, where firms are adaptive and profit-seeking agents entering into and exiting from the market according to their (lack of) profitability. Our firms are characterized by large and small sunk costs, respectively. They locate their offerings along a unimodal demand distribution over a one-dimensional product variety, with the distribution peak constituting the center and the tails standing for the peripheries. We found that large firms may first advance toward the most abundant demand spot, the market center, and release peripheral positions as predicted by extant dual market explanations. However, we also observed that large firms may then move back toward the market fringes to reduce competitive niche overlap in the center, triggering nonlinear resource occupation behavior. Novel results indicate that resource release dynamics depend on firm-level adaptive capabilities, and that a minimum scale of production for low sunk cost firms is key to the formation of the dual structure.


Evaluation of distribution coefficients and concentration ratios of (90)Sr and (137)Cs in the Techa River and the Miass River.
Empirical data on the behavior of radionuclides in aquatic ecosystems are needed for radioecological modeling, which is commonly used for predicting transfer of radionuclides, estimating doses, and assessing possible adverse effects on species and communities. Preliminary studies of radioecological parameters including distribution coefficients and concentration ratios, for (90)Sr and (137)Cs were not in full agreement with the default values used in the ERICA Tool and the RESRAD BIOTA codes. The unique radiation situation in the Techa River, which was contaminated by long-lived radionuclides ((90)Sr and (137)Cs) in the middle of the last century allows improved knowledge about these parameters for river systems. Therefore, the study was focused on the evaluation of radioecological parameters (distribution coefficients and concentration ratios for (90)Sr and (137)Cs) for the Techa River and the Miass River, which is assumed as a comparison waterbody. To achieve the aim the current contamination of biotic and abiotic components of the river ecosystems was studied; distribution coefficients for (90)Sr and (137)Cs were calculated; concentration ratios of (90)Sr and (137)Cs for three fish species (roach, perch and pike), gastropods and filamentous algae were evaluated. Study results were then compared with default values available for use in the well-known computer codes ERICA Tool and RESRAD BIOTA (when site-specific data are not available). We show that the concentration ratios of (137)Cs in whole fish bodies depend on the predominant type of nutrition (carnivores and phytophagous). The results presented here are useful in the context of improving of tools for assessing concentrations of radionuclides in biota, which could rely on a wider range of ecosystem information compared with the process limited the current versions of ERICA and RESRAD codes. Further, the concentration ratios of (90)Sr are species-specific and strongly dependent on Ca(2+) concentration in water. The universal characteristic allows us to combine the data of fish caught in the water with different mineralization by multiplying the concentration of Ca(2+). The concentration ratios for fishes were well-fitted by Generalized Logistic Distribution function (GLD). In conclusion, the GLD can be used for probabilistic modeling of the concentration ratios in freshwater fishes to improve the confidence in the modeling results. This is important in the context of risk assessment and regulatory.


Health effects related to wind turbine noise exposure: a systematic review.
Wind turbine noise exposure and suspected health-related effects thereof have attracted substantial attention. Various symptoms such as sleep-related problems, headache, tinnitus and vertigo have been described by subjects suspected of having been exposed to wind turbine noise. This review was conducted systematically with the purpose of identifying any reported associations between wind turbine noise exposure and suspected health-related effects. A search of the scientific literature concerning the health-related effects of wind turbine noise was conducted on PubMed, Web of Science, Google Scholar and various other Internet sources. All studies investigating suspected health-related outcomes associated with wind turbine noise exposure were included. Wind turbines emit noise, including low-frequency noise, which decreases incrementally with increases in distance from the wind turbines. Likewise, evidence of a dose-response relationship between wind turbine noise linked to noise annoyance, sleep disturbance and possibly even psychological distress was present in the literature. Currently, there is no further existing statistically-significant evidence indicating any association between wind turbine noise exposure and tinnitus, hearing loss, vertigo or headache. Selection bias and information bias of differing magnitudes were found to be present in all current studies investigating wind turbine noise exposure and adverse health effects. Only articles published in English, German or Scandinavian languages were reviewed. Exposure to wind turbines does seem to increase the risk of annoyance and self-reported sleep disturbance in a dose-response relationship. There appears, though, to be a tolerable level of around LAeq of 35 dB. Of the many other claimed health effects of wind turbine noise exposure reported in the literature, however, no conclusive evidence could be found. Future studies should focus on investigations aimed at objectively demonstrating whether or not measureable health-related outcomes can be proven to fluctuate depending on exposure to wind turbines.


The coccidian oocyst: a tough nut to crack!
Coccidian parasites are transmitted between hosts by the ingestion of food or water contaminated with oocysts, followed by the release of infectious sporozoites and invasion of the gastro-intestinal tract. In the external environment, sporozoites are protected from desiccation and chemical disinfection by the oocyst wall. This unique structure guarantees successful disease transmission and is as vital to the coccidian parasite as the exoskeleton is to insects--without it they would die. Here, we revisit the early work and combine it with newer molecular data to describe our present understanding of the coccidian oocyst wall.


Experiments and sensitivity coefficients analysis for multiphase flow model calibration of enhanced DNAPL dissolution.
Multiphase flow modeling is often used for the comparison and optimization of subsurface nonaqueous phase liquid (NAPL) remediation schemes. The calibration of such models is a challenging task due to the lack of detailed data describing the initial NAPL spatial distribution and the processes governing the fate and transport of NAPLs in porous media. In this study laboratory scale experiments were conducted to evaluate reagent-enhanced dense nonaqueous phase liquid (DNAPL) solubilization in saturated heterogeneous media. The DNAPL consisted of both pooled and residual saturation forms. To gain insight into the influence of various input parameters on effluent concentrations, the multiphase flow program was used to compute the sensitivity coefficients of key parameters, relating to the flow, flushing solution properties, soil parameters, NAPL distribution and mass transfer coefficient. The sensitivity coefficients were, in turn, used to aid in the model calibration and to underline the difficulties associated with the calibration of multiphase flow models, most notably the non-uniqueness of the calibration process when complete information is lacking. To alleviate this uncertainty and provide additional constraints, the conducted flushing experiments were jointly used to calibrate the multiphase flow model. The results of the model calibration suggest that the interphase mass transfer coefficient is dependent on the properties of the reagent aqueous solution used for DNAPL remediation, most notably the viscosity and interfacial tension.


Mechanical Properties and Simulated Wear of Provisional Resin Materials.
The purpose of this study was to determine flexural properties and erosive wear behavior of provisional resin materials. Three bis-acryl base provisional resins-1) Protemp Plus (PP), 2) Integrity (IG), 3) Luxatemp Automix Plus (LX)-and a conventional poly(methylmethacrylate) (PMMA) resin, UniFast III (UF), were evaluated. A resin composite, Z100 Restorative (Z1), was included as a benchmark material. Six specimens for each of the four materials were used to determine flexural strength and elastic modulus according to ISO Standard 4049. Twelve specimens for each material were used to examine wear using a generalized wear simulation model. The test materials were each subjected to wear challenges of 25,000, 50,000, 100,000, and 200,000 cycles in a Leinfelder-Suzuki (Alabama) wear simulator. The materials were placed in custom cylinder-shaped stainless-steel fixtures, and wear was generated using a cylindrical-shaped flat-ended stainless-steel antagonist in a slurry of nonplasticized PMMA beads. Wear (mean facet depth [μm] and volume loss [mm(3)]) was determined using a noncontact profilometer (Proscan 2100) with Proscan and AnSur 3D software. The laboratory data were evaluated using two-way analysis of variance (ANOVA; factors: 1) material and 2) cycles) followed by Tukey HSD post hoc test (α=0.05). The flexural strength ranged from 68.2 to 150.6 MPa, and the elastic modulus ranged from 2.0 to 15.9 GPa. All of the bis-acryl provisional resins (PP, IG, and LX) demonstrated significantly higher values than the PMMA resin (UF) in flexural strength and elastic modulus (p<0.05). However, there was no significant difference (p>0.05) in flexural properties among three bis-acryl base provisional resins (PP, IG, and LX). Z1 demonstrated significantly (p<0.05) higher flexural strength and elastic modulus than the other materials tested. The results for mean facet wear depth (μm) and standard deviations (SD) for 200,000 cycles were as follows: PP, 22.4 (5.0); IG, 51.0 (6.5); LX, 63.7 (4.5); UF, 70.5 (8.0); and Z1, 7.6 (1.2). Volume loss (mm(3)) and SDs for 200,000 cycles were as follows: PP, 0.311 (0.049); IG, 0.737 (0.074); LX, 0.919 (0.053); UF, 1.046 (0.127); and Z1, 0.111 (0.017). The two-way ANOVA showed a significant difference among materials (p<0.001) and number of cycles for both facet depth and volume loss. The post hoc test revealed differences (p<0.05) in wear values among the tested materials examined in this study. The findings provide valuable information regarding the flexural properties and the relative wear behavior of the provisional resins examined in this study.


Trends in fluoroquinolone resistance of Mycobacterium tuberculosis complex in a Taiwanese medical centre: 1995-2003.
Fluoroquinolones are being used more frequently for the treatment of multidrug-resistant (MDR) strains of Mycobacterium tuberculosis complex (MTB). This study was designed to determine the frequency of the emergence of fluoroquinolone-resistant strains in Taiwan and to assess whether this might be due to use of fluoroquinolones for treatment of patients with MDR or because of increased use of fluoroquinolones in the community for treatment of other infections. We also sought to determine whether there might be clonal spread of fluoroquinolone resistance. A total of 3497 clinical isolates of M. tuberculosis complex were obtained during 1995-2003, of which 141 were selected. They consisted of 62 isolates fully susceptible to four first-line drugs, 33 isolates resistant to rifampicin and isoniazid (MDR), and 46 isolates with a variety of any drug resistant patterns other than MDR (combination group). The MICs were determined for ciprofloxacin, ofloxacin and levofloxacin. An increase in the MIC90 and rates of resistance to ciprofloxacin, ofloxacin and levofloxacin were noted only in the MDR group. The rates were higher among strains isolated between 1998-2003 compared with those obtained between 1995-1997 (rate of resistance, 20% versus 7.7%; MIC > or = 4 mg/L versus 1-2 mg/L). Among the 10 fluoroquinolone-resistant isolates, five (50%) possessed mutations other than S95T in the gyrA gene. No gyrB mutation was found in any of the clinical isolates. These findings suggest that fluoroquinolone resistance is the result of treatment of patients with MDR strains rather than from use in the general community in Taiwan. The emergence of fluoroquinolone resistance among MDR strains reinforces the need for routine fluoroquinolone susceptibility testing whenever these drugs might be used.


Tocotrienol-rich fraction from palm oil prevents oxidative damage in diabetic rats.
This study was carried out to determine the effects of tocotrienol-rich fraction (TRF) (200 mg/Kg) on biomarkers of oxidative stress on erythrocyte membranes and leukocyte deoxyribonucleic acid (DNA) damage in streptozotocin (STZ)-induced diabetic rats. Male rats (n = 40) were divided randomly into four groups of 10: a normal group; a normal group with TRF; a diabetic group, and a diabetic group with TRF. Following four weeks of treatment, fasting blood glucose (FBG) levels, oxidative stress markers and the antioxidant status of the erythrocytes were measured. FBG levels for the STZ-induced diabetic rats were significantly increased (P <0.001) when compared to the normal group and erythrocyte malondialdehyde levels were also significantly higher (P <0.0001) in this group. Decreased levels of reduced glutathione and increased levels of oxidised glutathione (P <0.001) were observed in STZ-induced diabetic rats when compared to the control group and diabetic group with TRF. The results of the superoxide dismutase and glutathione peroxidase activities were significantly lower in the STZ-induced diabetic rats than in the normal group (P <0.001). The levels of DNA damage, measured by the tail length and tail moment of the leukocyte, were significantly higher in STZ-induced diabetic (P <0.0001). TRF supplementation managed to normalise the level of DNA damage in diabetic rats treated with TRF. Daily supplementation with 200 mg/Kg of TRF for four weeks was found to reduce levels of oxidative stress markers by inhibiting lipid peroxidation and increasing the levels of antioxidant status in a prevention trial for STZ-induced diabetic rats.


Analysis of human histone H2AZ deposition in vivo argues against its direct role in epigenetic templating mechanisms.
Chromatin is considered to be a principal carrier of epigenetic information due to the ability of alternative chromatin states to persist through generations of cell divisions and to spread on DNA. Replacement histone variants are novel candidates for epigenetic marking of chromatin. We developed a novel approach to analyze the chromatin environment of nucleosomes containing a particular replacement histone. We applied it to human H2AZ, one of the most studied alternative histones. We find that neither H2AZ itself nor other features of the H2AZ-containing nucleosome spread to the neighboring nucleosomes in vivo, arguing against a role for H2AZ as a self-perpetuating epigenetic mark.


Computation of coupled diffusion of oxygen, glucose and lactic acid in an intervertebral disc.
The present numerical study aims to investigate the disc nutrition and factors affecting it by evaluating the concentrations of oxygen, glucose and lactic acid in the disc while accounting for the coupling between these species via the pH level in the tissue and the nonlinear concentration-consumption (for glucose and oxygen) and concentration-production (for lactate) relations. The effects of changes in the endplate exchange area (EA) adjacent to the nucleus or the inner annulus for the transport of nutrients and in the disc geometry as well as tissue diffusivities under static compression loading on species concentrations are also studied. Moreover, alterations in solute diffusion following a central endplate fracture are investigated. An axisymmetric geometry with four distinct regions is considered. Supply sources are assumed at the outer annulus periphery and disc endplates. Coupling between different solutes, pH level, endplate disruptions (calcifications and fractures) and mechanical loads substantially influenced the distribution of nutrients throughout the disc as well as the magnitude and location of critical concentrations; maximum for the lactic acid and minimum for oxygen and glucose. In cases with loss of endplate permeability and/or disruptions therein, as well as changes in geometry and fall in diffusivity associated with fluid expression, the nutrient concentrations could fall to levels inadequate to maintain cellular activity or viability, thus initiating or accelerating disc degeneration.


Expression analysis of four endoglin missense mutations suggests that haploinsufficiency is the predominant mechanism for hereditary hemorrhagic telangiectasia type 1.
ENDOGLIN codes for a homodimeric membrane glycoprotein that interacts with receptors for members of the TGF-beta superfamily and is the gene mutated in the autosomal dominant vascular disorder hereditary hemorrhagic telangiectasia type 1 (HHT1). We recently demonstrated that functional endoglin was expressed at half levels on human umbilical vein endothelial cells (HUVECs) and peripheral blood activated monocytes from HHT1 patients. Two types of mutant protein were previously analyzed, the product of an exon 3 skip which was expressed as a transient intracellular species and prematurely truncated proteins that were undetectable in patient samples. Here we report the analysis of four proteins resulting from point mutations, with missense codons G52V and C53R in exon 2, W149C in exon 4 and L221P in exon 5. Metabolic labeling of activated monocytes from confirmed, clinically affected patients revealed reduced expression of fully processed normal endoglin in all cases. Pulse-chase analysis with HUVECs from a newborn with the C53R substitution indicated that mutant endoglin remained intracellular as a precursor form and did not impair processing of the normal protein. Biotinylation of cell surface proteins, metabolic labeling and pulse-chase analysis revealed that none of the engineered missense mutants was significantly expressed at the surface of COS-1 transfectants. Thus, these four HHT1 missense mutations lead to transient intracellular species which cannot interfere with normal endoglin function. These data suggest that haploinsufficiency, leading to reduced levels of one of the major surface glyco-proteins of vascular endothelium, is the predominant mechanism underlying the HHT1 phenotype.


Holographic memory system based on projection recording of computer-generated 1D Fourier holograms.
Utilization of computer generation of holographic structures significantly simplifies the optical scheme that is used to record the microholograms in a holographic memory record system. Also digital holographic synthesis allows to account the nonlinear errors of the record system to improve the microholograms quality. The multiplexed record of holograms is a widespread technique to increase the data record density. In this article we represent the holographic memory system based on digital synthesis of amplitude one-dimensional (1D) Fourier transform holograms and the multiplexed record of these holograms onto the holographic carrier using optical projection scheme. 1D Fourier transform holograms are very sensitive to orientation of the anamorphic optical element (cylindrical lens) that is required for encoded data object reconstruction. The multiplex record of several holograms with different orientation in an optical projection scheme allowed reconstruction of the data object from each hologram by rotating the cylindrical lens on the corresponding angle. Also, we discuss two optical schemes for the recorded holograms readout: a full-page readout system and line-by-line readout system. We consider the benefits of both systems and present the results of experimental modeling of 1D Fourier holograms nonmultiplex and multiplex record and reconstruction.


Epigenetic maintenance of topological domains in the highly rearranged gibbon genome.
The relationship between evolutionary genome remodeling and the three-dimensional structure of the genome remain largely unexplored. Here, we use the heavily rearranged gibbon genome to examine how evolutionary chromosomal rearrangements impact genome-wide chromatin interactions, topologically associating domains (TADs), and their epigenetic landscape. We use high-resolution maps of gibbon-human breaks of synteny (BOS), apply Hi-C in gibbon, measure an array of epigenetic features, and perform cross-species comparisons. We find that gibbon rearrangements occur at TAD boundaries, independent of the parameters used to identify TADs. This overlap is supported by a remarkable genetic and epigenetic similarity between BOS and TAD boundaries, namely presence of CpG islands and SINE elements, and enrichment in CTCF and H3K4me3 binding. Cross-species comparisons reveal that regions orthologous to BOS also correspond with boundaries of large (400-600 kb) TADs in human and other mammalian species. The colocalization of rearrangement breakpoints and TAD boundaries may be due to higher chromatin fragility at these locations and/or increased selective pressure against rearrangements that disrupt TAD integrity. We also examine the small portion of BOS that did not overlap with TAD boundaries and gave rise to novel TADs in the gibbon genome. We postulate that these new TADs generally lack deleterious consequences. Last, we show that limited epigenetic homogenization occurs across breakpoints, irrespective of their time of occurrence in the gibbon lineage. Overall, our findings demonstrate remarkable conservation of chromatin interactions and epigenetic landscape in gibbons, in spite of extensive genomic shuffling.


Efficacy and safety of weekly dalbavancin therapy for catheter-related bloodstream infection caused by gram-positive pathogens.
Catheter-related bloodstream infections (CR-BSIs) are associated with substantial mortality, prolongation of hospital stay, and increased cost of care. Dalbavancin, a new glycopeptide antibiotic with unique pharmacokinetic properties that have allowed clinical development of a weekly dosing regimen, possesses excellent activity against clinically important gram-positive bacteria, suggesting utility in the treatment of patients with CR-BSIs. A phase 2, open-label, randomized, controlled, multicenter study of 75 adult patients with CR-BSIs compared treatment with intravenous dalbavancin, administered as a single 1000-mg dose followed by a 500-mg dose 1 week later, with intravenous vancomycin, administered twice daily for 14 days. Gram-positive bacteria isolated in this study included coagulase-negative staphylococci (CoNS) and Staphylococcus aureus, including methicillin-resistant S. aureus (MRSA). Infected patients who received weekly dalbavancin (n=33) had an overall success rate (87.0%; 95% confidence interval [CI], 73.2%-100.0%) that was significantly higher than that of those who received vancomycin (n=34) (50.0%; 95% CI, 31.5%-68.5%). Adverse events and laboratory abnormalities were generally mild and were comparable for the 2 drugs. Dalbavancin thus appears to be an effective and well-tolerated treatment option for adult patients with CR-BSIs caused by CoNS and S. aureus, including MRSA.