Allotetraploid Mimulus sookensis are highly interfertile despite independent origins.
Polyploidy (whole-genome duplication) has contributed significantly to angiosperm evolution and diversification. To date, it has been found that most polyploids are the result of multiple formation events, which may contribute to genetic diversity and affect interfertility among polyploid lineages of independent origin. A recently discovered allotetraploid derivative of Mimulus guttatus and M. nasutus, Mimulus sookensis, is found throughout the valleys of western Oregon and Vancouver Island. Here, we analyse the patterns of nucleotide diversity at three chloroplast and six nuclear loci in M. guttatus, M. nasutus and M. sookensis, to gain insight into the formation of M. sookensis. By analysing the patterns of genetic variation seen in the diploid progenitors in comparison with the variation seen in M. sookensis, we are able to show that M. sookensis has recurrently formed. We also observed that most M. sookensis individuals are fixed heterozygotes at all of the nuclear loci examined, suggesting that duplicate gene loss is not extensive in M. sookensis. To assess the possibility that hybridization among M. sookensis has contributed to genetic diversity, we conducted crossing experiments within M. sookensis. We found that M. sookensis of independent origin are highly interfertile, suggesting that crossing barriers do not exist within M. sookensis, and that hybridization among M. sookensis may result in new recombinant genotypes. Together, the data suggest that although recurrent origins may be common, they can contribute to genetic diversity without contributing to reproductive isolation among independently arisen polyploid lineages.


What is the long-term outcome of a negative capsule endoscopy in patients with obscure gastrointestinal bleeding?
There are contradictory findings regarding long-term outcome in patients with obscure gastrointestinal bleeding and negative capsule endoscopy. Factors associated with rebleeding after a negative videocapsule are not entirely known. The aim of this study was to compare the rebleeding rate between negative and positive capsule endoscopy patients and to identify predictive factors for rebleeding in patients with negative findings. Consecutive patients with obscure gastrointestinal bleeding referred to a single center over a period of 5 years were identified. After exclusion of patients with a follow time < 6 months, 173 patients were included. Clinical information was retrospectively collected from medical records. Rebleeding was defined as evidence of melena/hematochezia, a drop in hemoglobin of ≥ 2 g/dL, or the need for transfusion 30 days after the index episode. The mean age was 61.7 years and 60% were female. The median follow up time was 27 months. Most patients were referred for occult gastrointestinal bleeding (67.1%) while 32.9% were referred for overt bleeding. More than 50% of the patients had negative capsule endoscopy. The rebleeding rate in negative capsule endoscopy is 16%, with a mean follow-up time of 25.8 months and is significantly lower than positive capsule endoscopy (16% vs. 30.2%, p = 0.02). Rebleeding after negative capsule endoscopy is higher in patients who need more transfusions of packet red blood cells before capsule endoscopy (3.0 vs. 0.9, p = 0.024) and have overt bleeding (46% vs. 13.9%, p = 0.03). In 53% of these patients, rebleeding occurs > 12 months after a negative capsule endoscopy. Patients with obscure gastrointestinal bleeding and a negative capsule endoscopy had a significantly lower rebleeding rate and can be safely followed. However, a higher transfusion of red blood cells previous to capsule endoscopy and an overt bleeding are associated with a higher rebleeding. So, it is reasonable to consider that these patients may benefit of at least one year of follow-up.


Electrical communication between glucose oxidase and electrodes mediated by phenothiazine-labeled poly(ethylene oxide) bonded to lysine residues on the enzyme surface.
A series of glucose oxidase (GOx) hybrids (GOx-phe-nothiazine-labeled poly(ethylene oxide) (PT-PEO)) capable of direct electrical communication with electrodes is synthesized by covalently modifying PT-PEO to lysine residues on the enzyme surface. The length of the PEO chain and the number of PT groups are systematically altered. After the PT-PEO modification, all the hybrids maintain more than 50% of enzyme activity relative to that of native GOx, although loss of the activity becomes greater with increasing PEO chain length. The catalytic current, i(cat), is observed at a potential more positive than 0.55 V after the addition of glucose, due to the intramolecular electron transfer (El) from reduced forms of flavin adenine dinucletide (FADH2/FADH) to PT+ that are electrogenerated at the electrode. The i(cat) value increases with the number of PT groups, indicating that most of the modified PT groups act as mediators. The magnitude of the i(cat) increase depends on the PEO chain length and reveals a maximum for PT-PEO with the molecular weight of 3,000. In contrast, the i(cat) is almost constant for GOx-2-(10-phenothiazyl)propionic acid (PT-PA) hybrids with more than two PT groups synthesized by covalently modifying PT-PA to surface lysines, indicating that only a few key PT groups function as mediators. The maximum rate constant (130 s(-1)) for the ET from FADH2/FADH to PT+ is obtained for the GOx hybrid modified with five PT-PEO groups with the molecular weight of 3,000.


Coccidioidomycosis Complement Fixation Titer Trends in the Age of Antifungals.
Coccidioidomycosis is associated with a broad spectrum of illness severity, ranging from asymptomatic or self-limited pulmonary infection to life-threatening manifestations of disseminated disease. Serologic studies before the widespread availability of antifungals established current understanding of serologic kinetics and dynamics. Chart histories and complement fixation (CF) titer trends were analyzed for 434 antifungal-treated coccidioidomycosis patients, who were classified by three infectious disease physicians as having either pulmonary uncomplicated coccidioidomycosis (PUC) (n = 248), pulmonary chronic coccidioidomycosis (PCC) (n = 64), disseminated coccidioidomycosis (DC) not including meningitis (n = 86), or coccidioidal meningitis (CM) (n = 36). The median maximal CF titers were 1:4 for PUC patients, 1:24 for PCC patients, 1:128 for DC patients, and 1:32 for CM patients. Approximately 25.4% of PUC patients, 6.2% of PCC patients, 2.3% of DC patients, and 8.3% of CM patients did not develop detectable titers during the study period. Maximal titers developed a mean of 31 days (95% confidence interval [CI], 13 to 50 days) after initial serologic positivity, with no significant differences between groups. Serologic recurrence occurred in 9% of PUC patients, 36% of PCC patients, 50% of DC patients, and 52% of CM patients. Median titer improvement rates were 91 days/dilution for PUC patients, 112 days/dilution for PCC patients, 136 days/dilution for DC patients, and 146 days/dilution for CM patients. Receiver operating characteristic (ROC) analysis revealed that CF testing retains moderate classification value for disseminated infections (area under the curve [AUC], 0.82 [95% CI, 0.78 to 0.87]) and complicated infections (AUC, 0.82 [95% CI, 0.77 to 0.86]). A suitable cutoff value for complicated infections is ≥1:32. Findings update serologic parameters that are relevant for clinical assessment of coccidioidomycosis patients in the triazole era.


Simultaneous Prediction of Wrist/Hand Motion via Wearable Ultrasound Sensing.
The ability to predict wrist and hand motions simultaneously is essential for natural controls of hand protheses. In this paper, we propose a novel method that includes subclass discriminant analysis (SDA) and principal component analysis for the simultaneous prediction of wrist rotation (pronation/supination) and finger gestures using wearable ultrasound. We tested the method on eight finger gestures with concurrent wrist rotations. Results showed that SDA was able to achieve accurate classification of both finger gestures and wrist rotations under dynamic wrist rotations. When grouping the wrist rotations into three subclasses, about 99.2 ± 1.2% of finger gestures and 92.8 ± 1.4% of wrist rotations can be accurately classified. Moreover, we found that the first principal component (PC1) of the selected ultrasound features was linear to the wrist rotation angle regardless of finger gestures. We further used PC1 in an online tracking task for continuous wrist control and demonstrated that a wrist tracking precision ( R2 ) of 0.954 ± 0.012 and a finger gesture classification accuracy of 96.5 ± 1.7% can be simultaneously achieved, with only two minutes of user training. Our proposed simultaneous wrist/hand control scheme is training-efficient and robust, paving the way for musculature-driven artificial hand control and rehabilitation treatment.


Mental health interventions in schools in low-income and middle-income countries.
Increasing enrolment rates could place schools in a crucial position to support mental health in low-income and middle-income countries. In this Review, we provide evidence for mental health interventions in schools in accordance with a public mental health approach spanning promotion, prevention, and treatment. We identified a systematic review for mental health promotion, and identified further prevention and treatment studies. Present evidence supports schools as places for promotion of positive aspects of mental health using a whole-school approach. Knowledge of effectiveness of prevention and treatment interventions is more widely available for conflict-affected children and adolescents. More evidence is needed to identify the many elements likely to be associated with effective prevention and treatment for children exposed to a range of adversity and types of mental disorders. Dissemination and implementation science is crucial to establish how proven effective interventions could be scaled up and implemented in schools.


Salinity Gradients for Sustainable Energy: Primer, Progress, and Prospects.
Combining two solutions of different composition releases the Gibbs free energy of mixing. By using engineered processes to control the mixing, chemical energy stored in salinity gradients can be harnessed for useful work. In this critical review, we present an overview of the current progress in salinity gradient power generation, discuss the prospects and challenges of the foremost technologies - pressure retarded osmosis (PRO), reverse electrodialysis (RED), and capacitive mixing (CapMix) and provide perspectives on the outlook of salinity gradient power generation. Momentous strides have been made in technical development of salinity gradient technologies and field demonstrations with natural and anthropogenic salinity gradients (for example, seawater-river water and desalination brine-wastewater, respectively), but fouling persists to be a pivotal operational challenge that can significantly ebb away cost-competitiveness. Natural hypersaline sources (e.g., hypersaline lakes and salt domes) can achieve greater concentration difference and, thus, offer opportunities to overcome some of the limitations inherent to seawater-river water. Technological advances needed to fully exploit the larger salinity gradients are identified. While seawater desalination brine is a seemingly attractive high salinity anthropogenic stream that is otherwise wasted, actual feasibility hinges on the appropriate pairing with a suitable low salinity stream. Engineered solutions are foulant-free and can be thermally regenerative for application in low-temperature heat utilization. Alternatively, PRO, RED, and CapMix can be coupled with their analog separation process (reverse osmosis, electrodialysis, and capacitive deionization, respectively) in salinity gradient flow batteries for energy storage in chemical potential of the engineered solutions. Rigorous techno-economic assessments can more clearly identify the prospects of low-grade heat conversion and large-scale energy storage. While research attention is squarely focused on efficiency and power improvements, efforts to mitigate fouling and lower membrane and electrode cost will be equally important to reduce levelized cost of salinity gradient energy production and, thus, boost PRO, RED, and CapMix power generation to be competitive with other renewable technologies. Cognizance of the recent key developments and technical progress on the different technological fronts can help steer the strategic advancement of salinity gradient as a sustainable energy source.


New subtype of spinocerebellar ataxia with altered vertical eye movements mapping to chromosome 1p32.
To provide clinical and genetic diagnoses for patients' conditions, it is important to identify and characterize the different subtypes of spinocerebellar ataxia (SCA). To clinically and genetically characterize a Spanish kindred with pure SCA presenting with altered vertical eye movements. DESIGN Family study of ambulatory patients. Electro-oculographic and genetics studies were performed in 2 referral university centers. Primary care institutional center in Spain. Thirty-six participants from a large Spanish kindred were clinically examined, and 33 family members were genetically examined. Detailed clinical data were obtained from 9 affected relatives. Two ataxic siblings and 2 asymptomatic family members were examined using an enhanced clinical protocol for a follow-up period of 7 years. High-density genome-wide single-nucleotide polymorphism arrays, along with microsatellite analysis, and genetic linkage studies were performed. Whole-exome sequencing was used for 2 affected relatives. For most patients, the initial symptoms included falls, dysarthria, or clumsiness followed by a complete cerebellar syndrome. For all 9 affected relatives, we observed altered vertical eye movements, as initial ocular signs for 3 of them and for the 2 asymptomatic family members, all having inherited the risk haplotype. Neuroimaging showed isolated cerebellar atrophy. Initial genome-wide linkage analysis revealed suggestive linkage to chromosome 1p32. Multipoint analysis and haplotype reconstruction further traced this SCA locus to a 0.66-cM interval flanked by D1S200 and D1S2742 (z(max) = 6.539; P < .0001). The causative mutation was unidentified by exome sequencing. We report a new subtype of SCA presenting in patients as slow progressing ataxia with altered vertical eye movements linked to a 11-megabase interval on 1p32. The Human Genome Nomenclature Committee has assigned this subtype of ataxia the designation SCA37.


Prions, prionoids and protein misfolding disorders.
Prion diseases are progressive, incurable and fatal neurodegenerative conditions. The term 'prion' was first nominated to express the revolutionary concept that a protein could be infectious. We now know that prions consist of PrPSc, the pathological aggregated form of the cellular prion protein PrPC. Over the years, the term has been semantically broadened to describe aggregates irrespective of their infectivity, and the prion concept is now being applied, perhaps overenthusiastically, to all neurodegenerative diseases that involve protein aggregation. Indeed, recent studies suggest that prion diseases (PrDs) and protein misfolding disorders (PMDs) share some common disease mechanisms, which could have implications for potential treatments. Nevertheless, the transmissibility of bona fide prions is unique, and PrDs should be considered as distinct from other PMDs.


Prognostic Significance and Clinicopathological Associations of COX-2 SNP in Patients with Nonsmall Cell Lung Cancer.
Background. To further improve the screening, diagnosis, and therapy of patients with nonsmall cell lung cancer (NSCLC) additional diagnostic tools are urgently needed. Gene expression of Cyclooxygenase-2 (COX-2) has been linked to prognosis in patients with NSCLC. The role of the COX-2 926G>C Single Nucleotide Polymorphism (SNP) in patients with NSCLC remains unclear. The aim of this study was to investigate the potential of the COX-2 926G>C SNP as a molecular marker in this disease. Methods. COX-2 926G>C SNP was analyzed in surgically resected tumor tissue of 85 patients with NSCLC using a PCR-based RFLP technique. Results. The COX-2 926G>C SNP genotypes were detected with the following frequencies: GG n = 62 (73%), GC n = 20 (23%), CC n = 3 (4%). There were no associations between COX-2 SNP genotype and histology, grading or gender detectable. COX-2 SNP was significantly associated with tumor stage (P = .032) and lymph node status (P = .016, Chi-square test). With a median followup of 85.9 months, the median survival was 59.7 months. There were no associations seen between the COX-2 SNP genotype and patients prognosis. Conclusions. The COX-2 926G>C SNP is detectable at a high frequency in patients with NSCLC. The COX-2 926G>C SNP genotype is not a prognostic molecular marker in this disease. However, patients with the GC or CC genotype seem more susceptible to lymph node metastases and higher tumor stage than patients with the GG genotype. The results suggest COX-2 926G>C SNP as a molecular marker for lymph node involvement in this disease.


APP duplication is sufficient to cause early onset Alzheimer's dementia with cerebral amyloid angiopathy.
We assessed the impact of amyloid precursor protein (APP) gene locus duplications in early onset Alzheimer's disease in a Dutch population-based sample. Using real-time PCR and an in-house-developed multiplex amplicon quantification assay, we identified a genomic APP duplication in 1 out of 10 multigenerational families segregating early onset Alzheimer's disease. In this family, cerebral amyloid angiopathy (CAA) coincided with this disease. The duplicated genomic region included no other genes than APP and extended maximally over 0.7 Mb. In a sample of 65 familial early onset patients, we observed the same APP genomic duplication in one patient (1.7%), while in 36 isolated patients duplications in the APP locus were absent. This indicated that APP locus duplications explained <2% of familial, non-autosomal dominant Alzheimer's disease and are an infrequent cause of de novo mutation. Our findings corroborated a recent French study, and indicated that investigating genomic duplications in the APP locus in families segregating Alzheimer's disease and CAA should be considered.


Targeted nanoparticles containing the proresolving peptide Ac2-26 protect against advanced atherosclerosis in hypercholesterolemic mice.
Chronic, nonresolving inflammation is a critical factor in the clinical progression of advanced atherosclerotic lesions. In the normal inflammatory response, resolution is mediated by several agonists, among which is the glucocorticoid-regulated protein called annexin A1. The proresolving actions of annexin A1, which are mediated through its receptor N-formyl peptide receptor 2 (FPR2/ALX), can be mimicked by an amino-terminal peptide encompassing amino acids 2-26 (Ac2-26). Collagen IV (Col IV)-targeted nanoparticles (NPs) containing Ac2-26 were evaluated for their therapeutic effect on chronic, advanced atherosclerosis in fat-fed Ldlr(-/-) mice. When administered to mice with preexisting lesions, Col IV-Ac2-26 NPs were targeted to lesions and led to a marked improvement in key advanced plaque properties, including an increase in the protective collagen layer overlying lesions (which was associated with a decrease in lesional collagenase activity), suppression of oxidative stress, and a decrease in plaque necrosis. In mice lacking FPR2/ALX in myeloid cells, these improvements were not seen. Thus, administration of a resolution-mediating peptide in a targeted NP activates its receptor on myeloid cells to stabilize advanced atherosclerotic lesions. These findings support the concept that defective inflammation resolution plays a role in advanced atherosclerosis, and suggest a new form of therapy.


Study on the effects of tablet colour in the treatment of anxiety states.
Forty-eight patients with anxiety states were treated with oxazepam (Serenid-D), which was administered in tablets of three different colours-red, yellow, and green. Every patient received one week's treatment with each colour, according to a random programme. A latin square design was used to ensure complete balance between the colours and between the weeks. The patients' symptoms were categorized and then assessed by both weekly physicians' ratings and daily self-rating, which showed close agreement. Colour preference was shown on both these scales in that symptoms of anxiety were most improved with green, whereas depressive symptoms appeared to respond best to yellow. Such colour preferences, however, did not reach levels of statistical significance, except for phobias as rated on the physicians' assessment.The results indicate that colour may play a part in the response to a drug.


Posttranscriptional regulation of the c-myb proto-oncogene in estrogen receptor-positive breast cancer cells.
We have determined that expression of the c-myb proto-oncogene is associated with estrogen receptor (ER) status and not with tumor progression in human breast epithelial cells. Analysis of normal, immortalized, nontumorigenic, and tumorigenic mammary epithelial cells showed that only ER+ tumor cell lines expressed readily detectable levels of c-myb mRNA and a Mr 75,000 protein that was the same size as the c-myb transcripts and protein products present in hematopoietic cells. In this report we show that c-myb mRNA and protein levels are down-regulated during estrogen withdrawal. A 20-fold increase in c-myb mRNA and protein expression was observed upon addition of beta-estradiol to the culture medium. Nuclear run-on transcription analyses showed that c-myb was transcribed at the same rate in the presence and absence of estrogen, suggesting that c-myb mRNA accumulation was regulated at a posttranscriptional level. To provide additional evidence that c-myb mRNA was dependent on ER expression, we examined c-myb mRNA levels in MCF-7 cells selected for resistance to antineoplastic drugs. c-myb expression was decreased only in cell lines that showed concomitant loss of ER expression. Moreover, c-myb mRNA was expressed and modulated by estrogen in ER-, MDA-MB-231 cells stably transfected with a human ER gene. When considered together, these data indicate that c-myb mRNA levels are regulated by estrogens and further suggest that this proto-oncogene plays a role in the biology of ER+ breast tumor cells.


The pharmacokinetic modelling of GI198745 (dutasteride), a compound with parallel linear and nonlinear elimination.
To characterize the pharmacokinetics of the dual 5alpha-reductase inhibitor GI198745 (dutasteride) to allow for more accurate predictions of GI198745 concentrations after different dosing schedules. In this randomized, single-blind, parallel group study, 32 healthy male volunteers received single oral doses of GI198745 ranging from 0.01 to 40 mg. Data were analysed by nonlinear mixed effects modelling using NONMEM where both linear and nonlinear pharmacokinetic models were examined. The time course of GI198745 serum concentrations indicated concentration dependent elimination, with the apparent half-life increasing with dose. Data were best described by a two-compartment model with first order absorption and parallel linear and nonlinear elimination pathways. Drug absorption was rapid, and was followed by a short distribution phase. A high volume of distribution (511 l) and a low linear clearance (0.58 l h(-1)) combined to give a half-life of up to 5 (1-7) weeks at high concentrations. As concentrations declined towards Km (0.96 ng ml(-1)), the proportion eliminated by the relatively rapid saturable elimination pathway, with a maximum clearance of 6.2 l h(-1), increased and the half-life reduced to about 3 days. The estimated inter individual variability for the linear clearance was high (CV = 70%). G1198745 pharmacokinetics are well described by a pharmacokinetic model with parallel linear and nonlinear elimination. Simulations using this model show that at daily doses of 0.1 mg the steady state drug concentrations, and the rate at which these are achieved, are mainly influenced by the nonlinear pathway, while at daily doses above 1 mg they are almost entirely influenced by the linear pathway.


The many faces of risk: a qualitative study of risk in outpatient involuntary treatment.
This study aimed to derive a conceptualized model of risk in outpatient involuntary psychiatric treatment that has utility and meaning for stakeholders. Thirty-eight participants-patients, caregivers, clinicians, and legal decision makers-participated in qualitative interviews about their experiences of outpatient involuntary psychiatric treatment. Interview data were analyzed by using a general inductive method. Six types of risk were identified: actual harm, social adversity, therapeutic outcome or compromised treatment, the system, interpersonal distress, and epistemic issues. There were overlaps between discourses on risk but variation in how different aspects of risk were emphasized by participant groups. A comprehensive model of risk contextualized to outpatient involuntary treatment is proposed. It incorporates domains of risk of harm to self or others, risk of social adversity, risk of excess distress, and risk of compromised treatment. This model may have instrumental value in the implementation and the scrutiny of risk-based mental health laws.


In memoriam: Celso-Ramon Garcia, M.D. (1922-2004), reproductive medicine visionary.
This article traces the career of Celso-Ramon Garcia (1922-2004), noted physician, educator, and internationally renowned pioneer in the field of reproductive endocrinology. His work helped to formulate oral contraceptives used by millions of women throughout the world. Garcia's research collaborators included Gregory Pincus and John Rock, who together finalized the landmark clinical data needed to secure initial FDA approval for "the pill" in 1960. In addition to Garcia's monumental work in contraceptive endocrinology, his scholarly interests encompassed physiology of the menopause, minimally invasive reproductive surgery, as well as psychological aspects of infertility. Closely identified with the University of Pennsylvania, Garcia was instrumental in establishing the first formal clinical program in reproductive biology and influenced countless young scientists whose training he supervised and mentored. His distinguished career was emblematic of the best of the medical profession, characterized by compassion, intellect, and a sincere desire to help others. Our manuscript outlines Garcia's wide range of interests, acknowledges his superior fund of knowledge, and honors his humanitarian spirit - all of which contributed to an impressive legacy of medical discoveries. The impact of Prof. Garcia's work will continue to be felt for many years.


Immunotoxins that target an oncogenic mutant epidermal growth factor receptor expressed in human tumors.
Human cancers arise from a series of mutations, many of which direct the expression of mutant proteins with altered functions. These aberrant proteins are attractive targets for new therapeutic agents. One such protein is a mutant epidermal growth factor receptor (EGFRvIII) that has an in-frame deletion near the NH2 terminus of its extracellular domain. This protein was first identified in human gliomas, but has also been shown to be present in lung and breast carcinomas. The deletion results in a receptor with constitutive tyrosine kinase activity that enhances the tumorigenicity of glioblastomas in vivo. The deletion also creates a tumor-specific cell-surface sequence at the deletion junction. Three specific anti-EGFRvIII mAbs have been isolated following immunization with a mixture of a deletion junction synthetic peptide and EGFRvIII as present on cell membranes. We have constructed immunotoxins by conjugating a modified version of Pseudomonas exotoxin A to these mAbs. Immunotoxins were tested on cells that had been transfected with cDNA for the EGFRvIII receptor and expressed receptor protein at 5 x 10(5) receptors/cell. All three immunotoxins were cytotoxic to these cells, with 50% inhibition of protein synthesis occurring in a 15-50 pM range. The immunotoxins specifically targeted EGFRvIII, as their cytotoxicity could be blocked by their respective free antibody. They showed little or no cytotoxicity to cells expressing high levels of normal epidermal growth factor receptors, demonstrating that they are able to discriminate between cells expressing the mutant receptor and those expressing the wild-type receptor. Immunotoxins targeted to mutant epidermal growth factor receptors are promising candidates for further development as tumor cell-specific therapeutic agents.


Fusion angle affects intervertebral adjacent spinal segment joint forces-Model-based analysis of patient specific alignment.
This study addresses the hypothesis that adjacent segment intervertebral joint loads are sensitive to the degree of lordosis that is surgically imposed during vertebral fusion. Adjacent segment degeneration is often observed after lumbar fusion, but a causative mechanism is not yet clearly evident. Altered kinematics of the adjacent segments and potentially nonphysiological mechanical joint loads have been implicated in this process. However, little is known of how altered alignment and kinematics influence loading of the adjacent intervertebral joints under consideration of active muscle forces. This study investigated these effects by simulating L4/5 fusions using kinematics-driven musculoskeletal models of one generic and eight sagittal alignment-specific models. Models featured different spinopelvic configurations but were normalized by body height, masses, and muscle properties. Fusion of the L4/5 segment was implemented in an in situ (22°), hyperlordotic (32°), and hypolordotic (8°) fashion and kinematic input parameters were changed accordingly based on findings of an in vitro investigation. Bending motion from upright standing to 45° forward flexion and back was simulated for all models in intact and fused conditions. Joint loads at adjacent levels and moment arms of spinal muscles experienced changes after all types of fusion. Hypolordotic configuration led to an increase of adjacent segment (L3/4) shear forces of 29% on average, whereas hyperlordotic fusion reduced shear by 39%. Overall, L4/5 in situ fusion resulted in intervertebral joint forces closest to intact loading conditions. An artificial decrease in lumbar lordosis (minus 14° on average) caused by an L4/5 fusion lead to adverse loading conditions, particularly at the cranial adjacent levels, and altered muscle moment arms, in particular for muscles in the vicinity of the fusion. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:131-139, 2017.


IV and oral fosfomycin pharmacokinetics in neonates with suspected clinical sepsis.
Fosfomycin has the potential to be re-purposed as part of a combination therapy to treat neonatal sepsis where resistance to current standard of care (SOC) is common. Limited data exist on neonatal fosfomycin pharmacokinetics and estimates of bioavailability and CSF/plasma ratio in this vulnerable population are lacking. To generate data informing the appropriate dosing of IV and oral fosfomycin in neonates using a population pharmacokinetic analysis of plasma and CSF data. The NeoFosfo study (NCT03453177) was a randomized trial that examined the safety and pharmacokinetics of fosfomycin comparing SOC versus SOC plus fosfomycin. Sixty-one neonates received fosfomycin (100 mg/kg IV q12h for 48 h) and then they converted to oral therapy at the same dose. Two plasma pharmacokinetic samples were taken following the first IV and oral doses, sample times were randomized to cover the whole pharmacokinetic profile and opportunistic CSF pharmacokinetic samples were collected. A population pharmacokinetic model was developed in NONMEM and simulations were performed. In total, 238 plasma and 15 CSF concentrations were collected. A two-compartment disposition model, with an additional CSF compartment and first-order absorption, best described the data. Bioavailability was estimated as 0.48 (95% CI = 0.347-0.775) and the CSF/plasma ratio as 0.32 (95% CI = 0.272-0.409). Allometric weight and postmenstrual age (PMA) scaling was applied; additional covariates included postnatal age (PNA) on clearance and CSF protein on CSF/plasma ratio. Through this analysis a population pharmacokinetic model has been developed that can be used alongside currently available pharmacodynamic targets to select a neonatal fosfomycin dose based on an infant's PMA, PNA and weight.


[Effects of tympanic membrane perforation on real-ear to coupler difference].
To investigate the effects of tympanic membrane perforation on real-ear to coupler difference (RECD) in adults. RECD was measured in 34 ears with dry tympanic membrane perforation, 34 normal subjects served as controls. There was significant difference (P<0.05) below 1 kHz (including 1 kHz) and in 4 kHz between experimental group and control group. For perforated ears, RECD was 3.5-15 dB less from 250 Hz to 1 kHz while the values of RECD was negative from 250 Hz to 750 Hz. A larger standard deviation was found in experimental group. The mean intra-subject standard deviation was 4.4 dB in experimental group as contrasted with 1.4 dB in control group. The equivalent ear canal volume appeared to be negatively correlated with the RECD reduction below 0.75 kHz and no correlation above 1 kHz. The more the equivalent ear canal volume, the smaller the RECD. It was found that the size of tympanic membrane perforation had no effect on RECD. It is recommended to perform real-ear measurement individually instead of using the age-appropriate average values, and to increase low frequency gain appropriately when fitting hearing aids for patients with dry tympanic membrane perforation.


Corticotropin releasing hormone (CRH) antagonist attenuates adjuvant induced arthritis: role of CRH in peripheral inflammation.
To determine whether a corticotropin releasing hormone (CRH) type 1-specific receptor antagonist, antalarmin, would alter the progression of inflammation in adjuvant induced arthritis (AIA) susceptible LEW/N rats by blocking local CRH mediated inflammatory responses or render AIA resistant F344/N rats more susceptible to AIA by blocking central CRH, thus reducing secretion of endogenous glucocorticoids. F344/N and LEW/N rats were assigned to either drug or vehicle groups and treated with 20 mg/kg antalarmin or vehicle alone BID for 25 days by intraperitoneal injection. Arthritis was induced in both antalarmin and vehicle treated LEW/N and F344/N rats by subcutaneous injections at the base of the tail of incomplete Freund's adjuvant containing 10 mg/ml heat killed Mycobacterium tuberculosis. Control F344/N and LEW/N rats were maintained on either antalarmin or vehicle. Chronic blockade of CRH-R1 with systemic antalarmin significantly ameliorated AIA in LEW/N rats, reducing the severity of inflammation in peripheral joints, evidenced by clinical and histopathology scores, and weight loss associated with disease onset. Antalarmin neither induced nor exacerbated arthritis expression in F344/N or LEW/N rats, despite suppression of levels of adjuvant induced corticosterone, the major antiinflammatory glucocorticoid in rats. Systemic blockade of CRH-RI appeared to predominantly block peripheral proinflammatory effects of immune CRH, rather than the systemic glucocorticoid mediated antiinflammatory effects of hypothalamic CRH. Results indicate that chronic treatment with a CRH antagonist attenuates progressive inflammation induced degeneration of synovia, cartilage, and bone in arthritic joints, suggesting that antalarmin may have therapeutic potential in treatment of human autoimmune and inflammatory disorders.


Eosinophil chemotaxis of supernatants from cultured Hodgkin's lymph node cells.
Cultured lymph node cell supernatants from five out of six cases of Hodgkin's disease were relatively more chemotactic for peripheral blood eosinophils than neutrophils. In contrast, supernatants from two cases of lymphocytic lymphoma and four nodes showing reactive hyperplasia were chemotactic for neutrophils but had little eosinophil chemotactic activity. In most instances the dgree of eosinophil infiltration observed histologically in the Hodgkin's lymph node correlated with elaboration of eosinophil chemotactic activity from the cultured cells. Following gel-filtration of three of the Hodgkin's lymphnode supernatants, four peaks of eosinophil chemotactic activity were demonstrated in each case. One of these corresponded in molecular size to th previously described eosinophil chemotactic factor of anaphylaxis (ECF-A). It is suggested that the eosinophil chemotactic activity of cultured lymph node cell supernatants may be on value in the diagnosis and classification of Hodgkin's disease.


First Report of Lasiodiplodia theobromae Causing Dieback of Aquilaria sinensis in Taiwan.
Incense trees (Aquilaria sinensis (Lour.) Gilg) belong to a plant family used for alternative medicine in China and the production of wood. In the summer of 2012, at a nursery in Niaosong district, Kaohsiung City, Taiwan, more than 30% of a total of 400 incense trees had dieback symptoms on twigs with leaves attached, leading to eventual death of the entire plant. Symptomatic twigs and trunk pieces from six trees were collected and discolored tissues were excised, surface sterilized in 0.5% sodium hypochlorite solution, rinsed in sterilized distilled water, dried on sterilized filter paper, and then placed in petri dishes containing 2% water agar (WA). The dishes were incubated at room temperature for 1 to 2 days to obtain fungal strains from diseased tissues. The hyphal tips from developing fungal colonies were transferred to potato dextrose agar (PDA, Difco) dishes and placed under UV light (12 h/day) at 30°C. The purified colonies were used as inoculum in the pathogenicity tests. Pathogenicity tests were performed on 2-month-old A. sinensis seedlings, each treatment had three plants. Each plant was wounded by removing bark of the twigs with a disinfected scalpel enough to place a mycelium plug (about 5 × 10 mm2) of 7-day-old fungal isolate on the wound. The inoculated area was wrapped with a wet paper towel and Parafilm. Control plants were treated with PDA plugs. The symptoms described above were observed on inoculated plants 4 to 8 days after inoculation whereas control plants did not show symptoms. Diseased twigs were cut and placed in a moist chamber 21 days after inoculation and conidia oozing from pycnidia were observed. The same fungal pathogen was reisolated from inoculated plants, but not from the control. To identify the pathogen, the fungus was cultured as described above. The colonies were initially white with green to gray aerial mycelium after 5 to 6 days and eventually turned darker. Immature conidia were hyaline and one-celled, but mature conidia were dark brown, two-celled, thin-walled, and oval-shaped with longitudinal striations. The average size of 100 conidia was 25.23 ± 1.97 × 13.09 ± 0.99 μm with a length/width ratio of 1.92. For the molecular identification, the internal transcribed spacer (ITS) region of ribosomal DNA was PCR amplified with primers ITS1 and ITS4 (2) and sequenced. The sequences were deposited in GenBank (Accession No. JX945583) and showed 99% identity to Lasiodiplodia theobromae (HM346871, GQ469929, and HQ315840). Hence, both morphological and molecular characteristics confirmed the pathogen as L. theobromae (Pat.) Griffon & Maubl (1). To the best of our knowledge, this is the first report of L. theobromae causing dieback on Incense tree. This disease threatens tree survival and may decrease the income of growers. References: (1) W. H. Ko et al. Plant Dis. 88:1383, 2004. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, New York, 1990.


Sulfo-quinovosyl-acyl-glycerol (SQAG), a eukaryotic DNA polymerase inhibitor and anti-cancer agent.
It was found that a class of sulfolipids known as sulfo-quinovosyl-acyl-glycerols (SQAGs) from ferns and algae are potent inhibitors of eukaryotic DNA polymerase alpha and beta and effective anti-neoplastic agents. In developing a procedure for the chemical synthesis of sulfolipids, many derivatives and stereoisomers of SQAGs have been obtained including sulfo-quinovosyl-monoacyl-glycerols (SQMGs) and sulfo-quinovosyl-diacyl-glycerols (SQDGs). This review describes studies on the structure-function relationship between synthetic SQAGs and DNA polymerase alpha and beta, and the relationship to cytotoxic activity. The major action was probably dependent on the fatty acid effect, which was reported previously, although each of the SQAGs was a much stronger inhibitor than just the fatty acid present in the SQAGs. The inhibitory effect could be influenced by the chain size of fatty acids in the SQAGs. The sulfonyl group in quinovose was also needed to inhibit the enzymes. Lineweaver-Burk plots of SQAGs indicated that DNA polymerase alpha was non-competitively inhibited, but the SQAGs were effective as antagonists of both the template-primer DNA-binding and the nucleotide substrate-binding of DNA polymerase beta. Based on these results, the molecular actions of SQAGs and drug design strategies for developing new anti-neoplastic agents were discussed.


Ageing and the response of plasma insulin, glucose and C-peptide concentrations to intravenous glucose in postmenopausal women.
1. Eighty-six apparently healthy postmenopausal women not receiving hormone replacement therapy were given an intravenous glucose tolerance test. Plasma glucose, insulin and C-peptide concentrations were determined in fasting and post-glucose challenge samples. 2. Using a multivariate regression model, with predictor variables of chronological age, menopausal age and body mass index, neither chronological age nor menopausal age correlated with fasting or post-challenge plasma glucose or C-peptide concentrations. In contrast, menopausal age was positively associated with fasting plasma insulin concentration (P = 0.038, model r2 = 0.107), insulin area (P = 0.01, model r2 = 0.236) and incremental insulin area (P = 0.024, model r2 = 0.243). This relationship could not be explained by differences in lifestyle variables of alcohol consumption, physical activity, previous duration of oral contraceptive usage, history of cigarette smoking or body mass index. 3. Our findings suggest that loss of ovarian function is associated with hyperinsulinaemia, possibly via alterations in the clearance of circulating insulin from the plasma. The hyperinsulinaemia observed may contribute to the increased risk of cardiovascular disease seen in postmenopausal women.


A comparison of Landsat 8, RapidEye and Pleiades products for improving empirical predictions of satellite-derived bathymetry.
Satellite derived bathymetry (SDB) enables rapid mapping of large coastal areas through measurement of optical penetration of the water column. The resolution of bathymetric mapping and achievable horizontal and vertical accuracies vary but generally, all SDB outputs are constrained by sensor type, water quality and other environmental conditions. Efforts to improve accuracy include physics-based methods (similar to radiative transfer models e.g. for atmospheric/vegetation studies) or detailed in-situ sampling of the seabed and water column, but the spatial component of SDB measurements is often under-utilised in SDB workflows despite promising results suggesting potential to improve accuracy significantly. In this study, a selection of satellite datasets (Landsat 8, RapidEye and Pleiades) at different spatial and spectral resolutions were tested using a log ratio transform to derive bathymetry in an Atlantic coastal embayment. A series of non-spatial and spatial linear analyses were then conducted and their influence on SDB prediction accuracy was assessed in addition to the significance of each model's parameters. Landsat 8 (30 m pixel size) performed relatively weak with the non-spatial model, but showed the best results with the spatial model. However, the highest spatial resolution imagery used - Pleiades (2 m pixel size) showed good results across both non-spatial and spatial models which suggests a suitability for SDB prediction at a higher spatial resolution than the others. In all cases, the spatial models were able to constrain the prediction differences at increased water depths.


One-lung ventilation with use of a double lumen tube in two dogs; when right might be wrong.
One-lung ventilation (OLV) is an anaesthetic technique utilised for improved visualisation and access of the surgical field during thoracoscopy. The authors present two cases that underwent OLV with use of a double lumen tube (DLT). The first case was intubated with endoscopic guidance for bronchial port intubation of the right mainstem bronchus. This dog experienced prolonged periods of intraoperative hypoxemia. Upon case review, it was suspected intubation of the mainstem bronchus resulted in occlusion of the right cranial lung lobe. In the second case, the DLT was placed bronchoscopically into the left mainstem bronchus with the aid of computed tomography (CT). Excellent intraoperative oxygenation was achieved. When DLTs are used in dogs, their anatomical differences from humans make them susceptible to additional lung occlusion and pulmonary shunting. Computed tomography is recommended as a fundamental addition to bronchoscopy for correct tube placement.


Neurodevelopmental and growth outcomes of extremely low birth weight infants after necrotizing enterocolitis.
Necrotizing enterocolitis (NEC) is a significant complication for the premature infant. However, subsequent neurodevelopmental and growth outcomes of extremely low birth weight (ELBW) infants with NEC have not been well described. We hypothesized that ELBW infants with surgically managed (SurgNEC) are at greater risk for poor neurodevelopmental and growth outcomes than infants with medically managed NEC (MedNEC) compared with infants without a history of NEC (NoNEC). The objective of this study was to compare growth, neurologic, and cognitive outcomes among ELBW survivors of SurgNEC and MedNEC with NoNEC at 18 to 22 months' corrected age. Multicenter, retrospective analysis was conducted of infants who were born between January 1, 1995, and December 31, 1998, and had a birth weight <1000 g in the National Institute of Child Health and Human Development Neonatal Research Network Registry. Neurodevelopment and growth were assessed at 18 to 22 months' postmenstrual age. chi2, t test, and logistic regression analyses were used. A total of 2948 infants were evaluated at 18 to 22 months, 124 of whom were SurgNEC and 121 of whom were MedNEC. Compared with NoNEC, both SurgNEC and MedNEC infants were of lower birth weight and had a greater incidence of late sepsis; SurgNEC but not MedNEC infants were more likely to have received a diagnosis of cystic periventricular leukomalacia and bronchopulmonary dysplasia and been treated with postnatal steroids. Weight, length, and head circumference <10 percentile at 18 to 22 months were significantly more likely among SurgNEC but not MedNEC compared with NoNEC infants. After correction for anthropometric measures at birth and adjusted age at follow-up, all growth parameters at 18 to 22 months for SurgNEC but not MedNEC infants were significantly less than for NoNEC infants. SurgNEC but not MedNEC was a significant independent risk factor for Mental Developmental Index <70 (odds ratio [OR]: 1.61; 95% confidence interval [CI]: 1.05-2.50), Psychomotor Developmental Index <70 (OR: 1.95; 95% CI: 1.25-3.04), and neurodevelopmental impairment (OR: 1.78; 95% CI: 1.17-2.73) compared with NoNEC. Among ELBW infants, SurgNEC is associated with significant growth delay and adverse neurodevelopmental outcomes at 18 to 22 months' corrected age compared with NoNEC. MedNEC does not seem to confer additional risk. SurgNEC is likely to be associated with greater severity of disease.


Chromosome mediated gene transfer of six DNA markers linked to the cystic fibrosis locus on human chromosome seven.
The DNA probes met and pJ3.11 are derived from loci on chromosome seven that are closely linked to, and probably flanking, the gene mutation causing cystic fibrosis (CF). We have shown that mitotic chromosomes from the cell line MNNG-HOS, which contains an activated met oncogene, can induce morphological transformation of mouse NIH-3T3 cells. Southern analysis of isolated transfectant cell lines with cloned dispersed repetitive human DNA sequences as probes demonstrated that several lines of transformed NIH 3T3 cells had stabley incorporated large segments of chromosome seven DNA. Southern blot analysis also demonstrated the presence of met, pJ3.11 and several other single copy sequences that had been previously localised to chromosome 7 within the transgenomes. In this way a further four genetic markers were shown to be physically linked to met, and thus to CF. These probes may prove useful in confirming the order of loci around CF and in the prenatal diagnosis of this common autosomal recessive disease.


The association between positive-negative reactions of informal caregivers of people with dementia and health outcomes in eight European countries: a cross-sectional study.
To describe the associations between positive and negative reactions of informal caregivers of people with dementia and health outcomes across eight European Countries. Caring for someone with dementia may have implications for the caregiver's own health and for the care recipient. These consequences could be associated with caregivers' reactions to the process of care. Association study based on cross-sectional data. Participants were people with dementia and their informal caregivers living at home or in long-term care institutions. Data were collected between November 2010-April 2012 using the Caregiver Reaction Assessment (with dimensions of self-esteem, lack of family support, financial problems, disrupted schedule and health problems) and associations were sought with informal caregiver burden, quality of life and psychological well-being and with dementia sufferers' neuropsychiatric symptoms, comorbidity and dependency in activities of daily living using correlation coefficients. Data from 2014 participants were used. Variability across countries was noted, as well as differences between care at home and in long-term care institutions. In general, self-esteem and lack of family support correlated with caregiver burden and psychological well-being. Associations were also found between disrupted schedule and caregiver burden, psychological well-being and quality of life. Health problems were clearly associated with caregiver burden, psychological well-being and quality of life. Study results support links between the reactions of informal caregivers of people with dementia and health outcomes. These may have implications in terms of how services are addressed.


Polycythemia vera: diagnosis, clinical course, and current management
Very important developments related to polycythemia vera (PV) have occurred during the last two decades. The discovery of Janus kinase (JAK) 2 mutations has changed both the diagnosis and clinical management of PV. Currently JAK2 molecular testing is essential in the diagnostic work-up and JAK2 mutation positivity is a major diagnostic criterion. The discovery of JAK2 mutations suggested that abnormal JAK-STAT signaling was a pivotal feature in the pathogenesis of Philadelphia-negative myeloproliferative neoplasms. This idea led to the development of JAK inhibitors. Currently ruxolitinib, a JAK1/JAK2 inhibitor, is also approved for PV patients with hydroxyurea resistance or intolerance. International collaborations have made it possible to describe disease characteristics and evolution better. Presently it is possible to quantify the symptomatic burden of the disease and to estimate prognosis. In spite of these developments, management of PV still largely depends on estimation of thromboembolic risk and trying to decrease the risk with or without cytoreductive medications. Different approaches have been proposed by international disease experts for the diagnosis, thromboembolic risk estimation, and drug selection. This paper aims to review clinical aspects of PV and propose a management algorithm. The authors also point to still unresolved questions and unmet needs in diagnosis and management.


Speciation analysis of aluminium(III) in natural waters and biological fluids by complexing with various catechols followed by differential pulse voltammetry detection.
The biological effects of aluminium have received much attention in recent years. Speciation of Al is of basic relevance as it concerns its reactivity and bioavailability. A differential pulse voltammetry (DPV) procedure is proposed for speciation analysis of Al(III) in natural waters and biological fluids using six catechols (L-dopa, dopamine, epinephrine, norepinephrine, caffeic acid and o-benzenediol) as electroactive ligands. The decrease of the DPV anodic peak current for each catechol ligand is linear with the increase of Al concentration. This speciation analysis idea is based on the measurement of the complexation capacity, namely, different affinities of Al(III) for catechols and organic ligands under two pH conditions. The labile monomeric Al fraction (mainly inorganic aluminium) is determined at pH 4.6, while the total monomeric Al fraction is determined at pH 8.5. The principle for Al(III) speciation analysis by an electrochemical method is discussed. This sensitive and simple fractionation method is successfully applied to the speciation analysis of Al in natural waters and the results agree well with those of Driscoll's method. The speciation analysis of Al in biological fluids is also explored and the results are compared with those obtained by ultrafiltration and dialysis. Compared with other speciation protocols the electrochemical method possesses some remarkable advantages: rapidity, high sensitivity, cheap instrumentation and a simple operation procedure.


Apoptosis-Inducing Factor (AIF) in Physiology and Disease: The Tale of a Repented Natural Born Killer.
Apoptosis-inducing factor (AIF) is a mitochondrial oxidoreductase that contributes to cell death programmes and participates in the assembly of the respiratory chain. Importantly, AIF deficiency leads to severe mitochondrial dysfunction, causing muscle atrophy and neurodegeneration in model organisms as well as in humans. The purpose of this review is to describe functions of AIF and AIF-interacting proteins as regulators of cell death and mitochondrial bioenergetics. We describe how AIF deficiency induces pathogenic processes that alter metabolism and ultimately compromise cellular homeostasis. We report the currently known AIFM1 mutations identified in humans and discuss the variability of AIFM1-related disorders in terms of onset, organ involvement and symptoms. Finally, we summarize how the study of AIFM1-linked pathologies may help to further expand our understanding of rare inherited forms of mitochondrial diseases.


Cytotoxicity and specificity of directed toxins composed of diphtheria toxin and the EGF-like domain of heregulin beta1.
As a step in the design of directed toxins, aimed at cells that overexpress HER receptors, particularly breast carcinoma cells, we studied the properties of a chimera of diphtheria toxin (DT) and heregulin beta1. The EGF-like growth hormone heregulin is a ligand for the HER3 and HER4 receptors and their heterodimers with HER2. The 60-residue EGF-like domain (hrg) of heregulin elicits a biological response and binds to these receptors primarily through its N terminus. We tested a fusion protein in which hrg replaces the C-terminal receptor-binding domain of DT (DT(389)hrg) and an alternative design in which this domain is fused to the N terminus of DT(389). Of those two constructs, the N-terminal fusion was not active as a directed toxin but elicited a growth response. The C-terminal fusion of hrg to DT(389) yielded a functional toxin and showed cell line specific cytotoxicity that is consistent with heregulin specificity. The binding of hrg to its cognate receptor is not impaired as shown by receptor activation, direct binding, and competition with free hrg. Cytotoxicity is dependent on high-affinity binding of DT(389)hrg to HER3 and HER4 receptors and is not mediated by HER2 overexpression alone. For those cell lines exhibiting high-affinity binding sites, the level of cytotoxicity correlates with the rate of internalization. Thus DT(389)hrg chimeras offer a possible avenue toward directed toxins against cells that overexpress HER receptors.


CHIP is a chaperone-dependent E3 ligase that ubiquitylates unfolded protein.
The ubiquitin-proteasome system catalyses the immediate destruction of misfolded or impaired proteins generated in cells, but how this proteolytic machinery recognizes abnormality of cellular proteins for selective elimination remains elusive. Here, we report that the C-terminus of Hsc70-interacting protein (CHIP) with a U-box domain is an E3 ubiquitin-ligase collaborating with molecular chaperones Hsp90 and Hsc70. Thermally denatured firefly luciferase was multiubiquitylated by CHIP in the presence of E1 and E2 (Ubc4 or UbcH5c) in vitro, only when the unfolded substrate was captured by Hsp90 or Hsc70 and Hsp40. No ubiquitylating activity was detected in CHIP lacking the U-box region. CHIP efficiently ubiquitylated denatured luciferase trapped by the C-terminal region of Hsp90, which contains a CHIP binding site. CHIP also showed self-ubiquitylating activity independent of target ubiquitylation. Our results indicate that CHIP can be regarded as 'a quality-control E3' that selectively ubiquitylates unfolded protein(s) by collaborating with molecular chaperones.


Dragon exploratory system on hepatitis C virus (DESHCV).
Even though hepatitis C virus (HCV) cDNA was characterized about 20 years ago, there is insufficient understanding of the molecular etiology underlying HCV infections. Current global rates of infection and its increasingly chronic character are causes of concern for health policy experts. Vast amount of data accumulated from biochemical, genomic, proteomic, and other biological analyses allows for novel insights into the HCV viral structure, life cycle and functions of its proteins. Biomedical text-mining is a useful approach for analyzing the increasing corpus of published scientific literature on HCV. We report here the first comprehensive HCV customized biomedical text-mining based online web resource, dragon exploratory system on hepatitis C virus (DESHCV), a biomedical text-mining and relationship exploring knowledge base was developed by exploring literature on HCV. The pre-compiled dictionaries existing in the dragon exploratory system (DES) were enriched with biomedical concepts pertaining to HCV proteins, their name variants and symbols to make it suitable for targeted information exploration and knowledge extraction as focused on HCV. A list of 32,895 abstracts retrieved via PubMed database using specific keywords searches related to HCV were processed based on concept recognition of terms from several dictionaries. The web query interface enables retrieval of information using specified concepts, keywords and phrases, generating text-derived association networks and hypotheses, which could be tested to identify potentially novel relationship between different concepts. Such an approach could also augment efforts in the search for diagnostic or even therapeutic targets. DESHCV thus represents online literature-based discovery resource freely accessible for academic and non-profit users via http://apps.sanbi.ac.za/DESHCV/ and its mirror site http://cbrc.kaust.edu.sa/deshcv/.


Mechanisms of WNK1 and WNK4 interaction in the regulation of thiazide-sensitive NaCl cotransport.
With-no-lysine (WNK) kinases are highly expressed along the mammalian distal nephron. Mutations in either WNK1 or WNK4 cause familial hyperkalemic hypertension (FHHt), suggesting that the protein products converge on a final common pathway. We showed previously that WNK4 downregulates thiazide-sensitive NaCl cotransporter (NCC) activity, an effect suppressed by WNK1. Here we investigated the mechanisms by which WNK1 and WNK4 interact to regulate ion transport. We report that WNK1 suppresses the WNK4 effect on NCC activity and associates with WNK4 in a protein complex involving the kinase domains. Although a kinase-dead WNK1 also associates with WNK4, it fails to suppress WNK4-mediated NCC inhibition; the WNK1 kinase domain alone, however, is not sufficient to block the WNK4 effect. The carboxyterminal 222 amino acids of WNK4 are sufficient to inhibit NCC, but this fragment is not blocked by WNK1. Instead, WNK1 inhibition requires an intact WNK4 kinase domain, the region that binds to WNK1. In summary, these data show that: (a) the WNK4 carboxyl terminus mediates NCC suppression, (b) the WNK1 kinase domain interacts with the WNK4 kinase domain, and (c) WNK1 inhibition of WNK4 is dependent on WNK1 catalytic activity and an intact WNK1 protein. These findings provide insight into the complex interrelationships between WNK1 and WNK4 and provide a molecular basis for FHHt.


Scalable active learning for multiclass image classification.
Machine learning techniques for computer vision applications like object recognition, scene classification, etc., require a large number of training samples for satisfactory performance. Especially when classification is to be performed over many categories, providing enough training samples for each category is infeasible. This paper describes new ideas in multiclass active learning to deal with the training bottleneck, making it easier to train large multiclass image classification systems. First, we propose a new interaction modality for training which requires only yes-no type binary feedback instead of a precise category label. The modality is especially powerful in the presence of hundreds of categories. For the proposed modality, we develop a Value-of-Information (VOI) algorithm that chooses informative queries while also considering user annotation cost. Second, we propose an active selection measure that works with many categories and is extremely fast to compute. This measure is employed to perform a fast seed search before computing VOI, resulting in an algorithm that scales linearly with dataset size. Third, we use locality sensitive hashing to provide a very fast approximation to active learning, which gives sublinear time scaling, allowing application to very large datasets. The approximation provides up to two orders of magnitude speedups with little loss in accuracy. Thorough empirical evaluation of classification accuracy, noise sensitivity, imbalanced data, and computational performance on a diverse set of image datasets demonstrates the strengths of the proposed algorithms.


Post-awakening Cortisol in Explosive Ordnance Disposal Technicians: A Replication Study in a Novel Population.
Amassing evidence suggests that post-awakening salivary cortisol rhythms (changes in cortisol throughout the day) may indicate health status. We previously established summary parameters for salivary cortisol in U.S. Navy Sea, Air, and Land and reported excellent parameter stability across 2 days of repeated sampling. To confirm the generalizability of our original findings to other military populations, we replicated the procedures of our prior report in another specialized military group. The purpose of this study was to (1) establish summary parameters of daily salivary cortisol rhythms, (2) evaluate summary parameter stability, and (3) assess the impact of sampling compliance in U.S. Navy Explosive Ordnance Disposal (EOD) technicians. Seventy active duty, male Explosive Ordnance Disposal technicians (mean ± SD age; 34.9 ± 6.5 years) self-collected saliva samples in a nondeployed setting on two consecutive weekdays at WAKE, WAKE + 30 min, WAKE + 60 min, 4 p.m., and 9 p.m. For salivary cortisol, we computed summary parameters, i.e., measures of magnitude and measures of pattern, and then evaluated their stabilities via correlational analyses and Cronbach's alpha (α). Compliance was objectively and subjectively evaluated using actigraphy and self-reported data, respectively. This research was conducted under a Naval Health Research Center Institutional Review Board approved study (NHRC.2015.0013). Average salivary cortisol concentrations increased at WAKE + 30 (mean ± SE reactivity; 48.9 + 6.8%) from WAKE, followed by a swift recovery at WAKE + 60. Approximately 10.9% of the group were classified as negative-responders (i.e., < 0% change from WAKE to WAKE + 30). The measures of magnitude demonstrated fair stability across two days (r value range: 0.37-0.45, ps < 0.01; α range: 0.54-0.62). Fifty-five percent of the sample was classified as compliant (defined as <15 min deviation from target sampling times) across both days. Compliance decreased to 31% when compliance criteria were refined to <5 min deviation; however, controlling for compliance did not overwhelmingly influence the more stable summary parameters of magnitude. These findings demonstrate a thorough replication, with some additional refinement, of our prior study, implying generalizability across diverse military populations. Study limitations include unsupervised saliva collection in a free-living setting, which is counterbalanced by ecological validity. The noninvasive salivary sampling protocol used in this study yields stable estimations of daily cortisol rhythms in specialized military men and is recommended as an operational health surveillance tool by which to monitor chronically stressed military members.


Cardiovascular involvement in patients with systemic sclerosis: insights from electromechanical characteristics of the heart.
Systemic sclerosis is a severe, life threatening autoimmune disease involving the skin and visceral organs, including the lungs, gastrointestinal tract, kidneys and heart. Cardiac involvement in systemic sclerosis, which is an important mortality predictor may involve myocardium, conduction system, vascular wall, pericardium, pulmonary vessels causing pulmonary hypertension. Thus, the detection of cardiac involvement is necessary in systemic sclerosis. With the more widespread use of novel non-invasive imaging techniques, the cardiac involvement is more frequently encountered in daily practice. Noninvasive evaluation can offer an advantage in early prediction of poor prognosis and give a chance to apply new therapeutic approaches for cardiac involvement. In this review, we intend to present cardiac involvement of systemic sclerosis and clinical diagnostic modalities, including electromechanical properties of the atrium and ventricles, for assessment this involvement.


Live-cell measurements of kinase activity in single cells using translocation reporters.
Although kinases are important regulators of many cellular processes, measuring their activity in live cells remains challenging. We have developed kinase translocation reporters (KTRs), which enable multiplexed measurements of the dynamics of kinase activity at a single-cell level. These KTRs are composed of an engineered construct in which a kinase substrate is fused to a bipartite nuclear localization signal (bNLS) and nuclear export signal (NES), as well as to a fluorescent protein for microscopy-based detection of its localization. The negative charge introduced by phosphorylation of the substrate is used to directly modulate nuclear import and export, thereby regulating the reporter's distribution between the cytoplasm and nucleus. The relative cytoplasmic versus nuclear fluorescence of the KTR construct (the C/N ratio) is used as a proxy for the kinase activity in living, single cells. Multiple KTRs can be studied in the same cell by fusing them to different fluorescent proteins. Here, we present a protocol to execute and analyze live-cell microscopy experiments using KTRs. We describe strategies for development of new KTRs and procedures for lentiviral expression of KTRs in a cell line of choice. Cells are then plated in a 96-well plate, from which multichannel fluorescent images are acquired with automated time-lapse microscopy. We provide detailed guidance for a computational analysis and parameterization pipeline. The entire procedure, from virus production to data analysis, can be completed in ∼10 d.


A near single crystalline TiO2 nanohelix array: enhanced gas sensing performance and its application as a monolithically integrated electronic nose.
We present high performance gas sensors based on an array of near single crystalline TiO(2) nanohelices fabricated by rotating oblique angle deposition (OAD). The combination of large surface-to-volume ratio, extremely small size (<30 nm) comparable to the Debye length, a near single crystallinity of TiO(2) nanohelices, together with the unique top-and-bottom electrode configuration hugely improves the H(2)-sensing performance, including ∼10 times higher response at 50 ppm, approximately a factor of 5 lower detection limit, and much faster response time than the conventional TiO(2) thin film devices. Beyond such remarkable performance enhancement, the excellent compatibility of the OAD method compared with the conventional micro-fabrication technology opens a new avenue for monolithic integration of high-performance chemoresistive sensors to fabricate a simple, low cost, reliable, yet fully functional electronic nose and multi-functional smart chips for in situ environmental monitoring.


Effect of Ras inhibition in hematopoiesis and BCR/ABL leukemogenesis.
Ras small GTPases are activated in many hematopoietic growth factor signaling and in hematological malignancies, but their role in hematopoiesis and leukemogenesis is not completely known. Here we examined the effect of Ras inhibition by a dominant negative mutant of Ras, N17 H-Ras, in adult hematopoiesis and in BCR/ABL leukemogenesis using the mouse bone marrow transduction and transplantation approach. We found that N17 H-Ras expression suppressed B- and T-lymphopoiesis and erythropoiesis. Interestingly, N17 H-Ras did not suppress myelopoiesis in the bone marrow, yet it greatly attenuated BCR/ABL-induced chronic myelogenous leukemia (CML)-like myeloproliferative disease. Most BCR/ABL + N17 H-Ras mice eventually developed pro-B lymphoblastic leukemia/lymphoma (B-ALL). These results suggest that Ras activation is essential for the development of lymphoid and erythroid cells but not myeloid cells and that Ras is a critical target of BCR/ABL in the pathogenesis of CML, but not B-ALL.


Shunt reduction with a fenestrated Amplatzer device.
In specific high-risk patients with congenital heart disease (CHD), a complete closure of an intracardiac defect/shunt is not possible for a variety of reasons. We report our experiences with an interventional approach for shunt-reduction using various modifications of a self-fabricated Amplatzer device in our institution. Retrospective analysis of patients with CHD having received an interventional partial shunt occlusion since 09/2005. Five patients, mean age 18.6(3.4-66) years, mean weight 36.4(14-102) kg, have been treated. In three patients (3.4, 3.9, 66 years) with an atrial septal defect (ASD) and a restrictive left ventricle (LV) (n = 1) or pulmonary arterial hypertension (PAH) (n = 2), respectively, an Amplatzer Septal Occluder (ASO) with a predilated (n = 2) or a presutured (n = 1) central hole was implanted. After successful immediate volume release in all, the balloon-dilated holes closed spontaneously during mid-term follow-up, pulmonary artery (PA) pressure and LV function remained normal. Two patients (2.7 and 17 years) with a Fontan circulation and severe cyanosis (saturation ≤80%) due to a large fenestration and elevated PA pressures received a partial occlusion of their shunt by implanting a centrally stented ASO or Amplatzer Vascular plug. After a follow-up of 31 and 39 months both stents remained patent under oral anticoagulation, oxygen saturation remained >85% with PA pressures unchanged, and both patients were in good clinical conditions. In patients with an ASD and significant PAH and/or restrictive LV physiology as well as in Fontan patients with a large surgically created fenestration but failing Fontan circulation, a partial closure with a self-fenestrated Amplatzer device can be a feasible and successful therapeutic option. Balloon-dilated fenestrations in the Amplatzer device tend to close spontaneously during follow-up. Nonresorbable sutures or stenting can ensure patency of the created holes.


Novel time-temperature and 'consume-within' indicator based on gas-diffusion.
The novel time-temperature indicator label comprises an ammonia sensitive indicator layer film pressed onto a second film, comprising an ammonia-generating, adhesive layer. When separated the blue-coloured indicator film reverts back to its original (ammonia free) yellow form at a controllable, temperature dependant rate. The labels are easily made and stored.


Chebyshev high-dimensional model representation (Chebyshev-HDMR) potentials: application to reactive scattering of H2 from Pt(111) and Cu(111) surfaces.
Gas phase and surface reactions involving polyatomic molecules are of central importance to chemical physics, and require accurately fit potential energy surfaces describing the interaction in their systems. Here, we propose a method for generating a High Dimensional Model Representation (HDMR) of a multidimensional potential energy surface (PES) and apply it to reactive molecule-surface scattering problems. In the HDMR treatment, all N degrees of freedom (DOF) of an N-dimensional PES are represented but only n < N are explicitly coupled. The HDMR is obtained from Chebyshev polynomial expansions for each degree of freedom, where expansion coefficients are efficiently evaluated using discrete cosine transform (DCT) algorithms and properties of Chebyshev polynomials. HDMR surfaces constructed for the reactive scattering of H2 from Pt(111) and Cu(111) are used in quantum dynamics simulations; the resultant state-resolved reaction and scattering probabilities are compared to those from simulations using full (6D) PESs and n-mode PESs from previous work. The results are encouraging, and suggest that this method may be applicable to "late barrier" reactive systems for which the previously-used n-mode representation fails.


Angiographic Characteristics of Pseudo-occlusion of the Internal Carotid Artery Before and After Stenting.
Various definitions of pseudo-occlusion and clinical outcomes after various treatments have been reported, but reports discussing the stenotic characteristics of pseudo-occlusion are rare. To analyze the angiographic characteristics of pseudo-occlusion of the internal carotid artery (ICA) before and after carotid artery stenting (CAS). We retrospectively reviewed the angiographic characteristics of 56 patients with pseudo-occlusion of the ICA treated with CAS. Angiographic changes were evaluated from digital subtraction angiography in terms of lesion characteristics of the stenosis and the diameter changes in the distal ICA before and after CAS. Fifty-six patients were successfully treated. Based on angiographic findings of ICA stenosis, 33 and 23 patients were classified into the single-channel and multiple-channel group, respectively. Regarding the diameter changes in the distal ICA after CAS, 31 cases were classified as immediate dilatation and the other 25 as restricted dilatation. Immediate dilatation of the distal ICA beyond the stent and that at a follow-up examination were observed significantly less frequently in the multiple-channel group than in the single-channel group. The use of multiple stents and stent occlusion at a follow-up examination were significantly more prevalent in the multiple-channel group than in the single-channel group. Stenotic lesions of pseudo-occlusion of the ICA were classified as single channel and multiple channel. Restricted dilatation of the distal ICA after CAS was more prevalent in the multiple-channel group. Because adverse events tended to be more frequent in the multiple-channel group than in the single-channel group, the indications for CAS should be determined carefully in multiple-channel patients. CAS, carotid artery stentingICA, internal carotid artery.


Why Effective Interventions Do Not Work for All Patients: Exploring Variation in Response to a Chronic Disease Management Intervention.
Half of all Americans have a chronic disease. Promoting healthy behaviors to decrease this burden is a national priority. A number of behavioral interventions have proven efficacy; yet even the most effective of these has high levels of nonresponse. In this study, we explore variation in response to an evidence-based community health worker (CHW) intervention for chronic disease management. We used a convergent parallel design that combined a randomized controlled trial with a qualitative process evaluation that triangulated chart abstraction, in-depth interviews and participant observation. Eligible patients lived in a high-poverty region and were diagnosed with 2 or more of the following chronic diseases: diabetes, obesity, hypertension or tobacco dependence. There were 302 patients in the trial, 150 of whom were randomly assigned to the CHW intervention. Twenty patients and their CHWs were included in the qualitative evaluation. We found minimal differences between responders and nonresponders by sociodemographic or clinical characteristics. A qualitative process evaluation revealed that health behavior change was challenging for all patients and most experienced failure (ie, gaining weight or relapsing with cigarettes) along the way. Responders seemed to increase their resolve after failed attempts at health behavior change, while nonresponders became discouraged and "shut down." Failure is a common and consequential aspect of health behavior change; a deeper understanding of failure should inform chronic disease interventions.


Distress vocalizations in maternally separated mouse pups: modulation via 5-HT(1A), 5-HT(1B) and GABA(A) receptors.
Young rodents emit ultrasonic vocalizations (USVs) when separated from their dams and littermates. Pharmacological agents that act on GABA(A) and/or 5-HT receptors and that alleviate anxiety in humans reduce the emission of these calls. 1) to investigate specific 5-HT1 receptor subtypes that modulate maternal separation-induced USVs in mice; 2) to assess the behavioral specificity of these effects; and 3) to compare 5-HT1 agonists with a positive neurosteroid modulator of the GABA(A) receptor complex. Seven-day old CFW mouse pups were isolated from their littermates and placed onto a 20 degrees C surface for 4 min. USVs between 30 and 80 kHz, grid crossing, and rectal temperature were measured in separate groups of mouse pups following subcutaneous administration of 5-HT1A and 5-HT1B receptor agonists and antagonists, the neurosteroid allopregnanolone, or the benzodiazepine midazolam. The 5-HT1A agonists (+)8-OH-DPAT (0.01-0.1 mg/kg) and flesinoxan (0.3-1.0 mg/kg), the selective 5-HT1B agonist CP-94,253 (0.03-30.0 mg/kg), and the mixed 5-HT1B/2C receptor agonist TFMPP (0.1-10.0 mg/kg) dose-dependently reduced USVs. These effects were reversed by the 5-HT1A receptor antagonist WAY 100,635 (0.1 mg/kg) or the 5-HT1B/D receptor antagonist GR 127935 (0.1 mg/kg). The effects of TFMPP were biphasic; low doses (i.e. 0.01 and 0.03 mg/kg) increased the rate of vocalization. Midazolam and allopregnanolone also reduced USVs. The highest doses of flesinoxan, (+)8-OH-DPAT, and allopregnanolone suppressed locomotion, whereas CP-94,253, TFMPP, and midazolam stimulated motor activity. These experiments confirm that agonists at the 5-HT1 receptors and a positive allosteric modulator of the GABA(A) receptor complex decrease maternal separation-induced USVs in mice, with 5-HT1B manipulations dissociating the effects on vocalizations from sedative effects.


The Fast Track intervention's impact on behaviors of despair in adolescence and young adulthood.
How to mitigate the dramatic increase in the number of self-inflicted deaths from suicide, alcohol-related liver disease, and drug overdose among young adults has become a critical public health question. A promising area of study looks at interventions designed to address risk factors for the behaviors that precede these -often denoted-"deaths of despair." This paper examines whether a childhood intervention can have persistent positive effects by reducing adolescent and young adulthood (age 25) behaviors that precede these deaths, including suicidal ideation, suicide attempts, hazardous drinking, and opioid use. These analyses test the impact and mechanisms of action of Fast Track (FT), a comprehensive childhood intervention designed to decrease aggression and delinquency in at-risk kindergarteners. We find that random assignment to FT significantly decreases the probability of exhibiting any behavior of despair in adolescence and young adulthood. In addition, the intervention decreases the probability of suicidal ideation and hazardous drinking in adolescence and young adulthood as well as opioid use in young adulthood. Additional analyses indicate that FT's improvements to children's interpersonal (e.g., prosocial behavior, authority acceptance), intrapersonal (e.g., emotional recognition and regulation, social problem solving), and academic skills in elementary and middle school partially mediate the intervention effect on adolescent and young adult behaviors of despair and self-destruction. FT's improvements to interpersonal skills emerge as the strongest indirect pathway to reduce these harmful behaviors. This study provides evidence that childhood interventions designed to improve these skills can decrease the behaviors associated with premature mortality.


Noninvasive evaluation of the upper extremity.
Vascular problems of the arm and hand can be assessed by a number of noninvasive modalities that are chosen on the basis of the history and physical examination. For suspected upper-extremity ischemia, we begin with SLPs and velocity-waveform analysis. The former test will define the extent and approximate location of the disease process, and subjective assessment of the waveform will further determine the degree and location of occlusive disease. When digital ischemia is suspected, the Doppler examination combined with intermittent compression of the radial and ulnar arteries is valuable for defining the variable arterial anatomy of the hand and the patency of the common and proper digital arteries. The extent of distal ischemia can be assessed by digital pressures. Duplex scanning has been found to be of value in determining the source of upper-extremity micro-emboli, in imaging suspected aneurysmal changes, and for evaluating arteriovenous fistulae and bypass grafts. Cold testing is used to confirm the diagnosis of Raynaud's disease after excluding proximal occlusive disease. When symptoms suggest intermittent arterial obstruction, arterial compression at the thoracic outlet is assessed by monitoring the arterial waveform during a series of maneuvers that change the anatomy of the outlet. Although a combination of IPG and venous Doppler examination accurately identifies venous occlusion, we routinely use duplex scanning in this setting. In addition to providing both anatomic and hemodynamic information about the subclavian vein, the jugular vein and the junction of the innominate vein can also be studied. Because of its ability to image in a coronal plane, MRI scanning is another nonivasive study that we have found useful for evaluation of venous anatomy and patency of the subclavian, jugular, and innominate veins. Venous thrombosis, often the first manifestation of subclavian vein compression at the thoracic inlet, is best evaluated using duplex scanning.


Noninvasive quantitation of drug concentration in prostate and seminal vesicles: improvement and validation with desipramine and aspirin.
The accessory glands of the male genital tract are the sites of several major health problems, including benign prostatic hyperplasia, prostate cancer, and human immunodeficiency virus (HIV) transmission. We aimed to validate and improve our noninvasive method for the quantitation of drug concentrations in these physiological subcompartments. Twenty-seven men were dosed with 100 mg desipramine (a weak base) and 975 mg aspirin (a weak acid) and ejaculated their semen in 1 pass across 5 compartments of a collection device 2.5 hours later. A Bayesian latent-variable model previously developed by our group was further advanced for the estimation of drug concentrations in prostate and seminal vesicles based on drug and biomarker concentrations in the split ejaculate. Under normality assumptions, desipramine concentration (with 95% credible intervals) in prostate and seminal vesicles were 27 (8.3-52) ng/mL and 7.6 (4.0-11) ng/mL, respectively; salicylate concentration in prostate and seminal vesicles were 2.0 (0.093-6.5) microg/mL, and 9.9 (8.2-12) microg/mL, respectively. The prostate-to-seminal vesicles concentration ratio was 0.20 (0.0087-0.75) for salicylate and 3.6 (0.91-9.9) for desipramine. We conclude that our quantitative analysis along with the split ejaculate method is sensitive, reproducible, and applicable for the assessment of pharmacokinetics of the accessory glands of the male genital tract.


Comparing alternative methods for holding virgin honey bee queens for one week in mailing cages before mating.
In beekeeping, queen honey bees are often temporarily kept alive in cages. We determined the survival of newly-emerged virgin honey bee queens every day for seven days in an experiment that simultaneously investigated three factors: queen cage type (wooden three-hole or plastic), attendant workers (present or absent) and food type (sugar candy, honey, or both). Ten queens were tested in each of the 12 combinations. Queens were reared using standard beekeeping methods (Doolittle/grafting) and emerged from their cells into vials held in an incubator at 34C. All 12 combinations gave high survival (90 or 100%) for three days but only one method (wooden cage, with attendants, honey) gave 100% survival to day seven. Factors affecting queen survival were analysed. Across all combinations, attendant bees significantly increased survival (18% vs. 53%, p<0.001). In addition, there was an interaction between food type and cage type (p<0.001) with the honey and plastic cage combination giving reduced survival. An additional group of queens was reared and held for seven days using the best method, and then directly introduced using smoke into queenless nucleus colonies that had been dequeened five days previously. Acceptance was high (80%, 8/10) showing that this combination is also suitable for preparing queens for introduction into colonies. Having a simple method for keeping newly-emerged virgin queens alive in cages for one week and acceptable for introduction into queenless colonies will be useful in honey bee breeding. In particular, it facilitates the screening of many queens for genetic or phenotypic characteristics when only a small proportion meets the desired criteria. These can then be introduced into queenless hives for natural mating or insemination, both of which take place when queens are one week old.


Self-enhancement of hepatitis C virus replication by promotion of specific sphingolipid biosynthesis.
Lipids are key components in the viral life cycle that affect host-pathogen interactions. In this study, we investigated the effect of HCV infection on sphingolipid metabolism, especially on endogenous SM levels, and the relationship between HCV replication and endogenous SM molecular species. We demonstrated that HCV induces the expression of the genes (SGMS1 and 2) encoding human SM synthases 1 and 2. We observed associated increases of both total and individual sphingolipid molecular species, as assessed in human hepatocytes and in the detergent-resistant membrane (DRM) fraction in which HCV replicates. SGMS1 expression had a correlation with HCV replication. Inhibition of sphingolipid biosynthesis with a hepatotropic serine palmitoyltransferase (SPT) inhibitor, NA808, suppressed HCV-RNA production while also interfering with sphingolipid metabolism. Further, we identified the SM molecular species that comprise the DRM fraction and demonstrated that these endogenous SM species interacted with HCV nonstructural 5B polymerase to enhance viral replication. Our results reveal that HCV alters sphingolipid metabolism to promote viral replication, providing new insights into the formation of the HCV replication complex and the involvement of host lipids in the HCV life cycle.


Structural Morphology of Molars in Large Mammalian Herbivores: Enamel Content Varies between Tooth Positions.
The distribution of dental tissues in mammalian herbivores can be very different from taxon to taxon. While grazers tend to have more elaborated and complexly folded enamel ridges, browsers have less complex enamel ridges which can even be so far reduced that they are completely lost. The gradient in relative enamel content and complexity of structures has so far not been addressed within a single species. However, several studies have noted tooth position specific wear rates in small mammals (rabbits, guinea pigs) which may be related to individual tooth morphology. We investigate whether differentiated enamel content by tooth position is also to be found in large herbivores. We use CT-scanning techniques to quantify relative enamel content in upper and lower molar teeth of 21 large herbivorous mammal species. By using a broad approach and including both perissodactyls and artiodactyls, we address phylogenetic intraspecific differences in relative enamel content. We find that enamel is highly unevenly distributed among molars (upper M1, M2, M3 and lower m1, m2, m3) in most taxa and that relative enamel content is independent of phylogeny. Overall, relative enamel content increases along the molar tooth row and is significantly higher in lower molars compared to upper molars. We relate this differential enamel content to prolonged mineralisation in the posterior tooth positions and suggest a compensatory function of m3 and M3 for functional losses of anterior teeth.


Enteric fever in Mediterranean north Africa.
Typhoid fever is endemic in the Mediterranean North African countries (Morocco, Algeria, Tunisia, Libya, and Egypt) with an estimated incidence of 10-100 cases per 100,000 persons. Outbreaks caused by Salmonella enterica serovar Typhi are common and mainly due to the consumption of untreated or sewage-contaminated water. Salmonella enterica Paratyphi B is more commonly involved in nosocomial cases of enteric fever in North Africa than expected and leads to high mortality rates among infants with congenital anomalies. Prevalence among travellers returning from this region is low, with an estimate of less than one per 100,000. Although multidrug resistant strains of Salmonella Typhi and Paratyphi are prevalent in this region, the re-emergence of chloramphenicol- and ampicillin-susceptible strains has been observed. In order to better understand the epidemiology of enteric fever in the Mediterranean North African region, population-based studies are needed. These will assist the health authorities in the region in preventing and controlling this important disease.


The effect of physiotherapy on knee joint extensor and flexor muscle strength after anterior cruciate ligament reconstruction using hamstring tendon.
The aim of this paper is to present the physiotherapeutic program employed at the Rehabilitation Centre for the College of Physiotherapy in Wrocław, Poland and its effectiveness by demonstrating the increase in strength of knee joint extensor and flexor muscles of patients after anterior cruciate ligament (ACL) reconstruction. Thirty-seven males participated in the physiotherapeutic program from the first week up to 8 months postoperatively. Each patient underwent an individual therapeutic program. Endoscopic reconstruction of a completely ruptured ACL was performed using Mitek's method (graft harvesting from flexor muscles). All patients previously underwent orthopedic and functional examinations, including measurements of the range of movement, knee and thigh circumference and strength of flexor and extensor muscles of the involved and uninvolved leg using a Biodex 3 System in both static and isokinetic modes. The outcome of the physiotherapeutic procedure, which is detailed in the paper, revealed a favorable effect of physiotherapy 6 months after ACL reconstruction. The observed 9% deficit in extensor muscle strength measured under isokinetic conditions of the involved knees compared with the uninvolved knees led us to conclude that the period of physiotherapy should be extended beyond 6 months for some patients. 1. Six months of physiotherapy following ACL reconstruction in males favorably affected muscle strength values of the involved and uninvolved knees under static and isokinetic conditions. 2. The application of individual loads during the sixth month of physiotherapy resulted in similar values of extensor and flexor muscle strength measured under static conditions, and flexor muscle strength measured under isokinetic conditions in involved and uninvolved knees.


Preference-based measures to obtain health state utility values for use in economic evaluations with child-based populations: a review and UK-based focus group assessment of patient and parent choices.
No current guidance is available in the UK on the choice of preference-based measure (PBM) that should be used in obtaining health-related quality of life from children. The aim of this study is to review the current usage of PBMs for obtaining health state utility values in child and adolescent populations, and to obtain information on patient and parent-proxy respondent preferences in completing PBMs in the UK. A literature review was conducted to determine which instrument is most frequently used for child-based economic evaluations and whether child or proxy responses are used. Instruments were compared on dimensions, severity levels, elicitation and valuation methods, availability of value sets and validation studies, and the range of utility values generated. Additionally, a series of focus groups of parents and young people (11-20 years) were convened to determine patient and proxy preferences. Five PBMs suitable for child populations were identified, although only the Health Utilities Index 2 (HUI2) and Child Heath Utility 9D (CHU-9D) have UK value sets. 45 papers used PBMs in this population, but many used non-child-specific PBMs. Most respondents were parent proxies, even in adolescent populations. Reported missing data ranged from 0.5 to 49.3%. The focus groups reported their experiences with the EQ-5D-Y and CHU-9D. Both the young persons' group and parent/proxy groups felt that the CHU-9D was more comprehensive but may be harder for a proxy to complete. Some younger children had difficulty understanding the CHU-9D questions, but the young persons' group nonetheless preferred responding directly. The use of PBMs in child populations is increasing, but many studies use PBMs that do not have appropriate value sets. Parent proxies are the most common respondents, but the focus group responses suggest it would be preferred, and may be more informative, for older children to self-report or for child-parent dyads to respond.


A life course approach to understanding associations between natural environments and mental well-being for the Danish blood donor cohort.
Natural environments have been associated with mental health benefits, but globally access to these benefits is threatened by urban development and densification. However, it remains unclear how natural environments relate to mental health and how consistent the association is across populations. Here we use a life-course approach with a population consisting of 66 194 individuals from the Danish Blood Donor Study (DBDS) to investigate the association between green and blue space (e.g. parks and lakes) and self-evaluated mental well-being. Green and blue space was identified from remotely-sensed images from the Landsat program, while mental well-being was based on the mental component score (MCS) calculated using the 12-item short form health survey. We use multivariate linear regression models and logistic regression models to quantify the associations. We adjust for additional environmental (urbanization, and air pollution) and lifestyle factors (smoking, body mass index, socioeconomic status, and physical activity) and specifically evaluate the role of physical activity and air pollution as possible mediating factors. We found a positive association between the MCS and current and childhood green space, and a non-significant association for current and childhood blue space. Adjusting for environmental and the other factors attenuated the effect sizes indicating that a broad range of factors determine mental well-being. Physical activity and air pollution were both associated with the MCS as possible mediators of green space associations. In addition, the odds for successfully completing tasks', seeing others, and feeling less downhearted increased with higher levels of green space, and the odds of feeling calm increased with higher levels of blue space. In conclusion, we found support for an association between green and, to less degree, blue space and mental well-being throughout different life stages. In addition, we found a positive association with individual indicators of mental well-being such as being productive, feeling less downhearted and calmer, and being social. The healthy blood donor effect and the bias towards urban residency may explain why we found smaller effect sizes between green and blue space and mental well-being for this generally healthy and resourceful cohort compared to previous studies.