Biofilm infections between Scylla and Charybdis: interplay of host antimicrobial peptides and antibiotics.
The aim of this study is to improve the anti-biofilm activity of antibiotics. We hypothesized that the antimicrobial peptide (AMP) complex of the host's immune system can be used for this purpose and examined the assumption on model biofilms. FLIP7, the AMP complex of the blowfly Calliphora vicina containing a combination of defensins, cecropins, diptericins and proline-rich peptides was isolated from the hemolymph of bacteria-challenged maggots. The complex interaction with antibiotics of various classes was studied in biofilm and planktonic cultures of Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Acinetobacter baumannii by the checkerboard method using trimethyl tetrazolium chloride cell viability and crystal violet biofilm eradication assays supplemented with microscopic analysis. We found that FLIP7 demonstrated: high synergy (fractional inhibitory concentration index <0.25) with meropenem, amikacin, kanamycin, ampicillin, vancomycin and cefotaxime; synergy with clindamycin, erythromycin and chloramphenicol; additive interaction with oxacillin, tetracycline, ciprofloxacin and gentamicin; and no interaction with polymyxin B. The interaction in planktonic cell models was significantly weaker than in biofilms of the same strains. The analysis of the dose-effect curves pointed to persister cells as a likely target of FLIP7 synergistic effect. The biofilm eradication assay showed that the effect also caused total destruction of S. aureus and E. coli biofilm materials. The effect allowed reducing the effective anti-biofilm concentration of the antibiotic to a level well below the one clinically achievable (2-3 orders of magnitude in the case of meropenem, ampicillin, cefotaxime and oxacillin). FLIP7 is a highly efficient host antimicrobial system helping antibiotics to overcome biofilm barriers through persisters' sensitization and biofilm material destruction. It is promising for the treatment of biofilm infections as an adjuvant of various small-molecule antibiotics.


Pulmonary hypoplasia presenting with recurrent wheezing in an infant.
Pulmonary hypoplasia is characterized by decrease in the number and size of pulmonary airways, alveoli and vessels. In autopsy, pulmonary hypoplasia is a major cause of death in neonates and infants. The disease is usually diagnosed in childhood period. Although it mimics lung parenchymal disease and other vascular abnormalities radiologically, it is easily recognized with computed tomography angiography and magnetic resonance angiography examinations. In 50% of patients, concomitant cardiovascular, neuromuscular, gastrointestinal tract, and urogenital anomalies are also available. There are two types of pulmonary hypoplasia: primary and secondary. Primary unilateral pulmonary hypoplasia may be asymptomatic and the tendency for bronchopulmonary infections is often increased in children. In this case report, a 22-month-old male patient characterized by recurrent infections and recurrent wheezes in infantile period, whose episodes of wheezing regressed after the pulmonectomy, was presented.


Genetic analysis of Gv1, a gene controlling transcription of endogenous murine polytropic proviruses.
Gv1 is a genetic locus that coordinately regulates the expression of multiple murine leukemia virus-related endogenous proviral sequences. A quantitative nuclease protection assay for typing Gv1 inheritance has been developed. Use of this assay demonstrates that Gv1 controls transcription of polytropic but not of modified polytropic endogenous proviruses. A combination of genetic techniques were used to map Gv1; these analyses demonstrate that Gv1 lies approximately 37 centimorgans from the centromeric end of mouse chromosome 13.


Patterns of evolutionary conservation of essential genes correlate with their compensability.
Essential genes code for fundamental cellular functions required for the viability of an organism. For this reason, essential genes are often highly conserved across organisms. However, this is not always the case: orthologues of genes that are essential in one organism are sometimes not essential in other organisms or are absent from their genomes. This suggests that, in the course of evolution, essential genes can be rendered nonessential. How can a gene become non-essential? Here we used genetic manipulation to deplete the products of 26 different essential genes in Escherichia coli. This depletion results in a lethal phenotype, which could often be rescued by the overexpression of a non-homologous, non-essential gene, most likely through replacement of the essential function. We also show that, in a smaller number of cases, the essential genes can be fully deleted from the genome, suggesting that complete functional replacement is possible. Finally, we show that essential genes whose function can be replaced in the laboratory are more likely to be non-essential or not present in other taxa. These results are consistent with the notion that patterns of evolutionary conservation of essential genes are influenced by their compensability-that is, by how easily they can be functionally replaced, for example through increased expression of other genes.


Protection of mesopore-adsorbed organic matter from enzymatic degradation.
Synthetic mesoporous alumina and silica minerals with uniform pore geometries, and their nonporous analogues, were used to test the role of mineral mesopores (2-50 nm diameter) in protecting organic matter from enzymatic degradation in soils and sediments. Dihydroxyphenylalanine (L-DOPA), a model humic compound, was irreversibly sorbed to both mineral types. The surface area-normalized adsorption capacity was greater for the mesoporous minerals relative to their nonporous analogues. The degradation kinetics of free and mineral-sorbed L-DOPA by the enzyme laccase was monitored in a closed cell via oxygen electrode. Relative to freely dissolved L-DOPA, nonporous alumina-sorbed substrate was degraded, on average, 90% more slowly and to a lesser extent (93%), likely due to laccase adsorption to alumina. In contrast, relative to free L-DOPA, degradation of nonporous silica-sorbed L-DOPA was enhanced by 20% on average. In the case of mesoporous alumina and silica-sorbed L-DOPA, the enzyme activity was 3-40 times lower than that observed for externally sorbed substrate (i.e., L-DOPA sorbed to nonporous minerals). These results provide strong evidence to support the viability of the mesopore protection mechanism for sequestration and preservation of sedimentary organic matter and organic contaminants. Nanopore adsorption/desorption phenomena may aid in explaining the slow degradation of organic contaminants in certain soils and sediments and may have implications for environmental remediation and biotechnological applications.


Integral transform analysis of radionuclide transport in variably saturated media using a physical non-equilibrium model: application to solid waste leaching at a uranium mining installation.
The Generalized Integral Transform Technique (GITT) was employed to simulate transient one-dimensional flow in variably saturated porous media, as well as radioactive waste transport within different layers (a solid waste pile, nearby soil, and a granular aquifer) towards the edge of a uranium mining installation under institutional control. Computational codes, written using the Mathematica software system, were implemented and tested for solution of the coupled advection-dispersion equations for an arbitrary number of daughter products within a radioactive chain migrating in saturated and unsaturated soil layers. The computer simulations were verified in great detail against results obtained using the built-in routine NDSolve of the Mathematica platform and the HYDRUS-1D software system. The present work reports the main results for 238U chain radionuclide transport using data extracted from a safety assessment of solid waste repositories at a uranium mining and milling installation in Caetité, state of Bahia, Brazil, operated by INB (Indústrias Nucleares do Brasil). Concentration evolutions of the various radionuclides obtained with the simulations were analyzed for five different cases to explore variations in the infiltration and recharge rates, the effect of assuming physical equilibrium or non-equilibrium transport conditions, and of different initial concentrations of some of the radionuclides.


Neural architecture underlying classification of face perception paradigms.
We present a novel strategy for deriving a classification system of functional neuroimaging paradigms that relies on hierarchical clustering of experiments archived in the BrainMap database. The goal of our proof-of-concept application was to examine the underlying neural architecture of the face perception literature from a meta-analytic perspective, as these studies include a wide range of tasks. Task-based results exhibiting similar activation patterns were grouped as similar, while tasks activating different brain networks were classified as functionally distinct. We identified four sub-classes of face tasks: (1) Visuospatial Attention and Visuomotor Coordination to Faces, (2) Perception and Recognition of Faces, (3) Social Processing and Episodic Recall of Faces, and (4) Face Naming and Lexical Retrieval. Interpretation of these sub-classes supports an extension of a well-known model of face perception to include a core system for visual analysis and extended systems for personal information, emotion, and salience processing. Overall, these results demonstrate that a large-scale data mining approach can inform the evolution of theoretical cognitive models by probing the range of behavioral manipulations across experimental tasks.


Deadlines at work and sleep quality. Cross-sectional and longitudinal findings among Danish knowledge workers.
Exposure to deadlines at work is increasing in several countries and may affect health. We aimed to investigate cross-sectional and longitudinal associations between frequency of difficult deadlines at work and sleep quality. Study participants were knowledge workers, drawn from a representative sample of Danish employees who responded to a baseline questionnaire in 2006 (n = 363) and a follow-up questionnaire in 2007 (n = 302). Frequency of difficult deadlines was measured by self-report and categorized into low, intermediate, and high. Sleep quality was measured with a Total Sleep Quality Score and two indexes (Awakening Index and Disturbed Sleep Index) derived from the Karolinska Sleep Questionnaire. Analyses on the association between frequency of deadlines and sleep quality scores were conducted with multiple linear regression models, adjusted for potential confounders. In addition, we used multiple logistic regression models to analyze whether frequency of deadlines at baseline predicted caseness of sleep problems at follow-up among participants free of sleep problems at baseline. Frequent deadlines were cross-sectionally and longitudinally associated with poorer sleep quality on all three sleep quality measures. Associations in the longitudinal analyses were greatly attenuated when we adjusted for baseline sleep quality. The logistic regression analyses showed that frequent deadlines at baseline were associated with elevated odds ratios for caseness of sleep problems at follow-up, however, confidence intervals were wide in these analyses. Frequent deadlines at work were associated with poorer sleep quality among Danish knowledge workers. We recommend investigating the relation between deadlines and health endpoints in large-scale epidemiologic studies.


Attitudes of patients with amyotrophic lateral sclerosis and their care givers toward assisted suicide.
Amyotrophic lateral sclerosis (ALS) is a neuromuscular disease that causes gradual paralysis, respiratory failure, and death, usually within three to five years after it has been diagnosed. Between 1995 and 1997, we surveyed patients with this disease in Oregon and Washington, as well as their family care givers, in order to determine their attitudes toward assisted suicide. Patients were considered to be willing to contemplate assisted suicide if they agreed with the statement, "Under some circumstances I would consider taking a prescription for a medicine whose sole purpose was to end my life," and disagreed with the statement, "I would never request or take a prescription for a medication whose sole purpose was to end my life." The Oregon Death with Dignity Act, which legalized physician-assisted suicide, was approved by Oregon voters in 1994 but did not go into effect until October 1997, after data collection for this study had been completed. Of 140 eligible persons with ALS, 100 (71 percent) agreed to participate in the study, as did 91 family care givers. The mean age of the patients with ALS was 54 years; the mean duration of illness since the diagnosis was 2.8 years. Fifty-six patients (56 percent) said they would consider assisted suicide, and 44 of the 56 agreed with the statement, "If physician-assisted suicide were legal, I would request a lethal prescription from a physician." One patient would have taken the medication immediately, and 36 would have kept it for future use. As compared with the patients who were opposed to assisted suicide, those who would consider it were more likely to be men, had a higher level of education, were less likely to be religious, had higher scores for hopelessness, and rated their quality of life as lower. In 66 of 91 instances (73 percent), care givers and patients had the same attitude toward assisted suicide. In Oregon and Washington, a majority of persons with ALS whom we surveyed would consider assisted suicide. Many would request a prescription for a lethal dose of medication well before they intended to use it.


Arteriovenous Fistula at the Craniocervical Junction Found After Cervical Laminoplasty for Ossification of the Posterior Longitudinal Ligament.
Ossification of the posterior longitudinal ligament (OPLL) is common in East Asia. Arteriovenous fistula at the craniocervical junction (CCJ-AVF), in contrast, is rare. As OPLL occurs most often in the cervical region, these 2 conditions can coexist in the cervical spinal canal of a single patient. We report a case of CCJ-AVF found after cervical laminoplasty (CLP) for OPLL. A 68-year-old man experienced progressive myelopathy due to cervical OPLL. Magnetic resonance imaging (MRI) revealed a high-intensity area inside the spinal cord. CLP was performed and his symptoms immediately improved. Three months after CLP, however, myelopathy recurred. MRI revealed an exacerbated and enlarged high-intensity area inside the cord from the medulla oblongata to the C4/5 level with a flow void around the cord. Left vertebral artery angiography revealed CCJ-AVF with ascending and descending draining veins. Direct surgery was performed to interrupt shunt flow into the draining veins. The patient's symptoms improved to a limited degree. In this case, increased pressure inside the spinal canal due to OPLL might have decreased the shunt flow of the CCJ-AVF. Thus, the venous congestion induced by CCJ-AVF might have been exacerbated after the pressure was removed by CLP. Magnetic resonance angiography screening could help detect concurrent CCJ-AVF and OPLL.


Intermittent testicular torsion.
The aim of this study is to highlight the clinical characteristics of patients with intermittent testicular torsion and draw attention to this underreported condition. Clinical and demographic data of all patients treated for intermittent testicular torsion from January 2007 to June 2015 were prospectively collected in a pro forma and analyzed. A diagnosis of intermittent torsion was made on the basis of recurrent scrotal pain, presence of abnormal testicular lie in otherwise normal testes, absence of urinary symptoms, and negative urine cultures. This diagnosis was confirmed by resolution of symptoms following bilateral orchidopexy. All patients had bilateral orchidopexy at the next operative day and were followed up for 12 months. Forty-five patients with a mean age of 20.9 years (±4.02) were seen within the study period. The left testis was more often involved than the right: 53.3% versus 37.8%. The condition was bilateral in 4 patients (8.9%). A total of 84 testes were evaluated. Horizontal lie was the most common anomaly causing intermittent testicular torsion 49%, followed by the clapper-bell deformity 27.5%. Patients experienced a mean of 2.8 (±1.74) acute pain episodes before testicular fixation. Bilateral orchidopexy resulted in resolution of symptoms and preservation of testicular volume. Horizontal lie of the testis is the most common cause of intermittent testicular torsion. The condition is more common on the left than the right testis and is predominantly unilateral. Intermittent testicular pain in the presence of abnormal testicular lie should warrant a diagnosis of intermittent testicular torsion. Early bilateral orchidopexy is efficacious.


Antibiotic effects of WP-0405, a thermo-setting ofloxacin gel, on methicillin-resistant Staphylococcus aureus keratitis in rabbits.
The chemotherapeutic effects and pharmacokinetics properties of WP-0405 (a thermo-setting in situ 0.3% ofloxacin-containing ophthalmic gel) and ofloxacin (a conventional 0.3% ofloxacin solution) on methicillin-resistant Staphylococcus aureus (MRSA) keratitis were compared in a rabbit model. The single-instillation pharmacokinetics of WP-0405 and ofloxacin in the cornea, aqueous humor, conjunctiva, and iris-ciliary body were determined in normal rabbit eyes. To compare the duration of antimicrobial action, WP-0405 or ofloxacin was instilled oncedaily in an early-treatment model of keratitis, and corneas were either removed immediately or 4 or 8 h postinstillation. In another experiment, WP-0405 was instilled two or three times daily to compare its antibiotic efficacy with three-times daily instillation of ofloxacin in the same early-treatment model of keratitis; corneas were then removed after determining the extent of the abscess area. In another experiment, WP-0405 was instilled four or eight times daily to compare its effects with eight-times daily instillation of ofloxacin in a late-treatment model of keratitis, and corneas were removed. The number of viable bacteria in the corneas was determined in all experiments. Cmax and AUC0- in tissues treated with WP-0405 were 1.5-3.4-fold and 1.8-2.9-fold greater than those treated with ofloxacin, respectively. WP-0405 significantly reduced the number of viable bacteria for up to 8 h after a single instillation. WP-0405 not only significantly reduced the number of viable bacteria, but also the size of the abscess area at the same frequency of instillation. When compared to ofloxacin, WP-0405 exhibited an approximately equivalent antibiotic effect, with fewer administrations. As a result of its pharmacokinetics, WP-0405 had a more potent, longer-acting antibiotic effect than did ofloxacin. Furthermore, because of its lower required instillation frequency, which would improve patient compliance, WP-0405 has great potential therapeutic benefits.


The immune system: a weapon of mass destruction invented by evolution to even the odds during the war of the DNAs.
Living systems operate under interactive selective pressures. Populations have the ability to anticipate the future by generating a repertoire of elements that cope with new selective pressures. If the repertoire of such elements were transcendental, natural selection could not operate because any one of them would be too rare. This is the problem that vertebrates faced in order to deal with a vast number of pathogens. The solution was to invent an immune system that underwent somatic evolution. This required a random repertoire that was generated somatically and divided the antigenic universe into combinatorials of determinants. As a result, it became virtually impossible for pathogens to escape recognition but the functioning of such a repertoire required two new regulatory mechanisms: 1) a somatic discriminator between Not-To-Be-Ridded ('Self') and To-Be-Ridded ('Non-self') antigens, and 2) a way to optimize the magnitude and choice of the class of the effector response. The principles governing this dual regulation are analyzed in the light of natural selection.


Laboratory and clinical development of single keyhole umbilical nephrectomy.
To describe our initial experience with single keyhole nephrectomy in a porcine model and in human subjects. Eight nonsurvival laparoscopic nephrectomies were performed in 4 female farm pigs. In 3 renal units, the laparoscopic nephrectomy was performed via a novel single 25-mm trocar, while the remaining 5 nephrectomies were performed using one 10-mm and two 5-mm adjacent trocars. Articulating laparoscopic graspers, conventional endoshears, clips, and a stapler were used for dissection. Three human subjects underwent a single keyhole umbilical laparoscopic nephrectomy with similar instrumentation. Indications for nephrectomy included chronic infection in a nonfunctioning kidney in 2 patients, and a 4.5-cm enhancing renal mass in the other patient. Single keyhole nephrectomy was successfully completed in all 8 porcine renal units and in all 3 human subjects. The mean operative time for the porcine nephrectomies was 49 minutes (range, 20 to 85), with a mean blood loss of 20 mL (range, 5 to 100). Incision size ranged from 3 to 5 cm. The mean operative time for the human nephrectomy cases was 133 minutes (range, 90 to 160). Estimated blood loss was 30 mL, and the kidneys were extracted through a solitary 2 to 4.5 cm periumbilical incision. There were no perioperative complications, and all 3 patients were discharged on hospital day 2. Keyhole umbilical nephrectomy utilizing articulating laparoscopic instrumentation to facilitate triangulation is feasible. We demonstrate safe and successful completion both in a porcine model and in the 3 human patients. Future studies will need to assess the benefits of single-access surgery in comparison to conventional laparoscopy.


A new mechanism regulating the initiation of allergic airway inflammation.
The earliest immune events induced by allergens are poorly understood, yet are likely essential to understanding how allergic inflammation is established. We sought to describe the earliest signaling events activated by allergen and determine their significance to allergic inflammation. A fungal-associated allergenic proteinase (FAP) or ovalbumin was administered once intranasally to wild-type mice to determine their ability to induce allergy-associated genes and initiate allergic lung inflammation. Mice deficient in recombinase activating gene 1, C3a, the C3a anaphylatoxin receptor, and MyD88 were challenged similarly to understand the requirement of these molecules and T and B cells for allergic inflammation. Adoptive T-cell transfer experiments were further performed to determine whether signal transducer and activator of transcription 6 (STAT6) was required for cell recruitment and allergic inflammation. FAP, but not ovalbumin, induced eosinophilic airway inflammation and lung IL-4 production in the absence of adaptive immune cells after the transcriptional induction of allergy-specific airway chemokines. Allergen-mediated chemokine secretion and innate allergic lung inflammation occurred in the absence of STAT6, recombinase activating gene 1, C3a, C3a anaphylatoxin receptor, Toll-like receptor 4, and MyD88 but required intact proteinase activity. Furthermore, FAP induced recruitment of T(H)2 cells and eosinophils to lungs independently of STAT6, which was previously thought to be required for T(H)2 cell homing. FAP induces allergic lung inflammation through a previously unrecognized innate immune signaling mechanism. These findings reveal a new paradigm for understanding how allergic inflammation begins and suggest novel possibilities for the prevention and treatment of allergic diseases, such as asthma.


B-Cell Therapies in Multiple Sclerosis.
B cells play a vital function in multiple sclerosis (MS) pathogenesis through an array of effector functions. All currently approved MS disease-modifying therapies alter the frequency, phenotype, or homing of B cells in one way or another. The importance of this mechanism of action has been reinforced with the successful development and clinical testing of B-cell-depleting monoclonal antibodies that target the CD20 surface antigen. Ocrelizumab, a humanized anti-CD20 monoclonal antibody, was approved by the Food and Drug Administration (FDA) in March 2017 after pivotal trials showed dramatic reductions in inflammatory disease activity in relapsing MS as well as lessening of disability progression in primary progressive MS. These and other clinical studies place B cells at the center of the inflammatory cascade in MS and provide a launching point for development of therapies that target selective pathogenic B-cell populations.


Nickel Sequestration by the Host-Defense Protein Human Calprotectin.
The human innate immune protein calprotectin (CP, S100A8/S100A9 oligomer, calgranulin A/calgranulin B oligomer, MRP-8/MRP-14 oligomer) chelates a number of first-row transition metals, including Mn(II), Fe(II), and Zn(II), and can withhold these essential nutrients from microbes. Here we elucidate the Ni(II) coordination chemistry of human CP. We present a 2.6-Å crystal structure of Ni(II)- and Ca(II)-bound CP, which reveals that CP binds Ni(II) ions at both its transition-metal-binding sites: the His3Asp motif (site 1) and the His6 motif (site 2). Further biochemical studies establish that coordination of Ni(II) at the hexahistidine site is thermodynamically preferred over Zn(II). We also demonstrate that CP can sequester Ni(II) from two human pathogens, Staphylococcus aureus and Klebsiella pneumoniae, that utilize this metal nutrient during infection, and inhibit the activity of the Ni(II)-dependent enzyme urease in bacterial cultures. In total, our findings expand the biological coordination chemistry of Ni(II)-chelating proteins in nature and provide a foundation for evaluating putative roles of CP in Ni(II) homeostasis at the host-microbe interface and beyond.


Meta-analysis of the relation between European and American smokeless tobacco and oral cancer.
Smokeless tobacco is often referred to as a major contributor to oral cancer. In some regions, especially Southeast Asia, the risk is difficult to quantify due to the variety of products, compositions (including non-tobacco ingredients) and usage practices involved. In Western populations, the evidence of an increased risk in smokeless tobacco users seems unclear, previous reviews having reached somewhat differing conclusions. We report a detailed quantitative review of the evidence in American and European smokeless tobacco users, and compare our findings with previous reviews and meta-analyses. Following literature review a meta-analysis was conducted of 32 epidemiological studies published between 1920 and 2005 including tests for homogeneity and publication bias. Based on 38 heterogeneous study-specific estimates of the odds ratio or relative risk for smokeless tobacco use, the random-effects estimate was 1.87 (95% confidence interval 1.40-2.48). The increase was mainly evident in studies conducted before 1980. No increase was seen in studies in Scandinavia. Restricting attention to the seven estimates adjusted for smoking and alcohol eliminated both heterogeneity and excess risk (1.02; 0.82-1.28). Estimates also varied by sex (higher in females) and by study design (higher in case-control studies with hospital controls) but more clearly in studies where estimates were unadjusted, even for age. The pattern of estimates suggests some publication bias. Based on limited data specific to never smokers, the random-effects estimate was 1.94 (0.88-4.28), the eight individual estimates being heterogeneous and based on few exposed cases. Smokeless tobacco, as used in America or Europe, carries at most a minor increased risk of oral cancer. However, elevated risks in specific populations or from specific products cannot definitely be excluded.


Geomicrobiology of sublacustrine thermal vents in Yellowstone Lake: geochemical controls on microbial community structure and function.
Yellowstone Lake (Yellowstone National Park, WY, USA) is a large high-altitude (2200 m), fresh-water lake, which straddles an extensive caldera and is the center of significant geothermal activity. The primary goal of this interdisciplinary study was to evaluate the microbial populations inhabiting thermal vent communities in Yellowstone Lake using 16S rRNA gene and random metagenome sequencing, and to determine how geochemical attributes of vent waters influence the distribution of specific microorganisms and their metabolic potential. Thermal vent waters and associated microbial biomass were sampled during two field seasons (2007-2008) using a remotely operated vehicle (ROV). Sublacustrine thermal vent waters (circa 50-90°C) contained elevated concentrations of numerous constituents associated with geothermal activity including dissolved hydrogen, sulfide, methane and carbon dioxide. Microorganisms associated with sulfur-rich filamentous "streamer" communities of Inflated Plain and West Thumb (pH range 5-6) were dominated by bacteria from the Aquificales, but also contained thermophilic archaea from the Crenarchaeota and Euryarchaeota. Novel groups of methanogens and members of the Korarchaeota were observed in vents from West Thumb and Elliot's Crater (pH 5-6). Conversely, metagenome sequence from Mary Bay vent sediments did not yield large assemblies, and contained diverse thermophilic and nonthermophilic bacterial relatives. Analysis of functional genes associated with the major vent populations indicated a direct linkage to high concentrations of carbon dioxide, reduced sulfur (sulfide and/or elemental S), hydrogen and methane in the deep thermal ecosystems. Our observations show that sublacustrine thermal vents in Yellowstone Lake support novel thermophilic communities, which contain microorganisms with functional attributes not found to date in terrestrial geothermal systems of YNP.


Influence of the Near Molecular Vicinity on the Temperature Regulated Fluorescence Response of Poly(N-vinylcaprolactam).
A series of new fluorescent dye bearing monomers, including glycomonomers, based on maleamide and maleic esteramide was synthesized. The dye monomers were incorporated by radical copolymerization into thermo-responsive poly(N‑vinyl-caprolactam) that displays a lower critical solution temperature (LCST) in aqueous solution. The effects of the local molecular environment on the polymers' luminescence, in particular on the fluorescence intensity and the extent of solvatochromism, were investigated below as well as above the phase transition. By attaching substituents of varying size and polarity in the close vicinity of the fluorophore, and by varying the spacer groups connecting the dyes to the polymer backbone, we explored the underlying structure⁻property relationships, in order to establish rules for successful sensor designs, e.g., for molecular thermometers. Most importantly, spacer groups of sufficient length separating the fluorophore from the polymer backbone proved to be crucial for obtaining pronounced temperature regulated fluorescence responses.


Effect of nerve growth factor on the proliferation in newborn bovine testicular Sertoli cells.
Nerve growth factor (NGF) has been proved to play important roles in male reproductive physiology, but the molecular mechanisms of NGF action remain unclear. In this study, the effects of NGF on the growth of newborn bovine testicular Sertoli (NBS) cells and the related signaling pathways were investigated. The NBS cells were treated in vitro with NGF (100 ng/mL) for 18 h. The expression levels of cell proliferation related genes, INHBB, and cytoplasmic specialization related gene were determined using real-time PCR and Western blot. The roles of PI3K/AKT and MAPK/ERK pathways in NGF-induced cell proliferation were investigated. It was found that NGF regulates proliferation and function of NBS cells via its receptor NTRK1 by activating the PI3K/ATK and MAPK/ERK signaling pathways. The study will help to further understand the role of NGF in male reproduction and provide new therapeutic targets for reproductive dysfunctions in male animals.


Actinorhodin biosynthesis: structural requirements for post-PKS tailoring intermediates revealed by functional analysis of ActVI-ORF1 reductase.
Actinorhodin (ACT) produced by Streptomyces coelicolor A3(2) is an aromatic polyketide antibiotic, whose basic carbon skeleton is derived from type II polyketide synthase (PKS). Although an acyl carrier protein (ACP) serves as an anchor of nascent intermediates during chain elongation in the type II PKS complex, it generally remains unknown when an ACP-free intermediate is released from the complex to post-PKS modification ("tailoring") steps. In ACT biosynthesis, a stereospecific ketoreductase (RED1) encoded by actVI-ORF1 reduces the 3beta-keto group of a proposed bicyclic intermediate to an (S) secondary alcohol. The bicyclic intermediate is formed from the steps of PKS and its closely associated enzymes and lies at the interface toward ACT-tailoring steps. To clarify whether RED1 recognizes the ACP-bound bicyclic intermediate or the ACP-free bicyclic intermediate, recombinant RED1 was purified for enzymatic characterization. RED1 was heterologously expressed in Escherichia coli and purified using Ni-chelate and gel filtration column chromatographies to homogeneity in soluble form. Enzymatic studies in vitro on RED1 with synthetic analogues, in place of an unstable bicyclic intermediate, showed that RED1 recognizes 3-oxo-4-naphthylbutyric acid (ONBA) as a preferred substrate and not its N-acetylcysteamine thioester. This strongly suggests that RED1 recognizes ACP-free bicyclic beta-keto acid as the first committed intermediate of tailoring steps. Kinetic studies of RED1 showed high affinity with ONBA, consistent with the requirement for an efficient reduction of a labile beta-keto carboxylic acid. Interestingly, the methyl ester of ONBA acted as a competitive inhibitor of RED1, indicating the presence of strict substrate recognition toward the terminal acid functionality.


Beta-galactosidase is inactivated by intermolecular disulfide bonds and is toxic when secreted to the periplasm of Escherichia coli.
The wild-type LamB-LacZ hybrid protein inhibits the export machinery upon induction when assayed by biochemical and genetic techniques, a phenotype referred to as hybrid protein jamming. This hybrid protein also renders cells sensitive to growth in the presence of the inducer maltose, presumably because of the jamming. We constructed a new version of this fusion by adding alkaline phosphatase, encoded by phoA, to the C terminus of the LamB-LacZ hybrid protein. This tripartite protein, LamB-LacZ-PhoA, is as toxic to cells as the hybrid LamB-LacZ; however, it does not jam at temperatures greater than 33 degrees C. Extreme C-terminal sequences of LacZ function as a critical folding domain and are therefore responsible for stabilizing the LacZ structure. To determine if this region of LacZ is important for jamming, we recombined a late nonsense mutation (X90) onto the hybrid construct. We found the toxicity of this new hybrid, LamB-LacZX90, to be nearly identical to that of the full-length protein, but it also does not jam the secretion machinery. This suggests that jamming is caused by LacZ folding. We found no inhibition of secretion in the tripartite and X90 fusion strains at 37 degrees C, suggesting that the toxicity of the new fusions is novel. Under these conditions, the tripartite and X90 fusion proteins form disulfide-bonded aggregates with high molecular weights in the periplasm. Accordingly, we believe that LacZ disrupts some essential function(s) in the periplasm.


Diesel spray image segmentation with a likelihood ratio test.
To characterize the macroscopic behavior of Diesel sprays and to validate and extend for current high-pressure injection systems the correlations existent in the literature, it is necessary to determine the spray geometry accurately, at least in terms of spray tip penetration and cone angle. These parameters are measured by analyzing Diesel spray images and are highly sensitive to the correct edge determination. An algorithm for segmentation of color images based on a likelihood ratio test is presented. This algorithm is compared with others available in the literature and has been validated, even for adverse experimental conditions. The experimental facilities, optical layouts, and image-processing algorithms are described.


Functional Connectivity Alterations between Networks and Associations with Infant Immune Health within Networks in HIV Infected Children on Early Treatment: A Study at 7 Years.
Although HIV has been shown to impact brain connectivity in adults and youth, it is not yet known to what extent long-term early antiretroviral therapy (ART) may alter these effects, especially during rapid brain development in early childhood. Using both independent component analysis (ICA) and seed-based correlation analysis (SCA), we examine the effects of HIV infection in conjunction with early ART on resting state functional connectivity (FC) in 7 year old children. HIV infected (HIV+) children were from the Children with HIV Early Antiretroviral Therapy (CHER) trial and all initiated ART before 18 months; uninfected children were recruited from an interlinking vaccine trial. To better understand the effects of current and early immune health on the developing brain, we also investigated among HIV+ children the association of FC at 7 years with CD4 count and CD4%, both in infancy (6-8 weeks) and at scan. Although we found no differences within any ICA-generated resting state networks (RSNs) between HIV+ and uninfected children (27 HIV+, 18 uninfected), whole brain connectivity to seeds located at RSN connectivity peaks revealed several loci of FC differences, predominantly from seeds in midline regions (posterior cingulate cortex, paracentral lobule, cuneus, and anterior cingulate). Reduced long-range connectivity and increased short-range connectivity suggest developmental delay. Within the HIV+ children, clinical measures at age 7 years were not associated with FC values in any of the RSNs; however, poor immune health during infancy was associated with localized FC increases in the somatosensory, salience and basal ganglia networks. Together these findings suggest that HIV may affect brain development from its earliest stages and persist into childhood, despite early ART.


Evaluation of urine and serum metabolites in miniature schnauzers with calcium oxalate urolithiasis.
To evaluate underlying causes of calcium oxalate urolithiasis, 24-hour excretion of urine metabolites was measured in 6 Miniature Schnauzers that formed calcium oxalate (CaOx) uroliths during periods when they were fed a standard diet and during periods when food was withheld. Serum concentrations of parathyroid hormone and 1,25-dihydroxyvitamin D also were evaluated. Serum calcium concentrations were normal in all 6 affected Miniature Schnauzers; however, during diet consumption, mean 24-hour urinary excretion of calcium was significantly (P = 0.025) higher than calcium excretion when food was withheld. In 1 dog, urinary calcium excretion was lower during the period of food consumption, compared with the period when food was withheld. Compared with clinically normal Beagles, Miniature Schnauzers that formed CaOx uroliths excreted significantly greater quantities of calcium when food was consumed (P = 0.0004) and when food was withheld (P = 0.001). Miniature Schnauzers that formed CaOx uroliths excreted significantly less oxalate than clinically normal Beagles during fed (P = 0.028) and nonfed (P = 0.004) conditions. Affected Miniature Schnauzers also excreted abnormally high quantities of uric acid. Excretion of citrate was not different between Miniature Schnauzers with CaOx urolithiasis and clinically normal Beagles. In 5 of 6 Miniature Schnauzers with CaOx urolithiasis, concentrations of serum parathyroid hormone were similar to values from age- and gender-matched Miniature Schnauzers without uroliths. The concentration of serum parathyroid hormone in 1 dog was greater than 4 times the mean concentration of clinically normal Miniature Schnauzers. Mean serum concentrations of 1,25-dihydroxyvitamin D in Miniature Schnauzers with calcium oxalate urolithiasis were similar to concentrations of clinically normal Miniature Schnauzers.


Epidemiology of Coxiella burnetii infection in Africa: a OneHealth systematic review.
Q fever is a common cause of febrile illness and community-acquired pneumonia in resource-limited settings. Coxiella burnetii, the causative pathogen, is transmitted among varied host species, but the epidemiology of the organism in Africa is poorly understood. We conducted a systematic review of C. burnetii epidemiology in Africa from a "One Health" perspective to synthesize the published data and identify knowledge gaps. We searched nine databases to identify articles relevant to four key aspects of C. burnetii epidemiology in human and animal populations in Africa: infection prevalence; disease incidence; transmission risk factors; and infection control efforts. We identified 929 unique articles, 100 of which remained after full-text review. Of these, 41 articles describing 51 studies qualified for data extraction. Animal seroprevalence studies revealed infection by C. burnetii (≤13%) among cattle except for studies in Western and Middle Africa (18-55%). Small ruminant seroprevalence ranged from 11-33%. Human seroprevalence was <8% with the exception of studies among children and in Egypt (10-32%). Close contact with camels and rural residence were associated with increased seropositivity among humans. C. burnetii infection has been associated with livestock abortion. In human cohort studies, Q fever accounted for 2-9% of febrile illness hospitalizations and 1-3% of infective endocarditis cases. We found no studies of disease incidence estimates or disease control efforts. C. burnetii infection is detected in humans and in a wide range of animal species across Africa, but seroprevalence varies widely by species and location. Risk factors underlying this variability are poorly understood as is the role of C. burnetii in livestock abortion. Q fever consistently accounts for a notable proportion of undifferentiated human febrile illness and infective endocarditis in cohort studies, but incidence estimates are lacking. C. burnetii presents a real yet underappreciated threat to human and animal health throughout Africa.


Measuring inhibitory processes for alcohol-related attentional biases: introducing a novel attentional bias measure.
Attentional biases for alcohol related information (AB) have often been reported for heavy drinkers. These attentional biases have been found to have predictive value regarding relapse in abstaining alcoholics. Similarly impaired inhibitory processes have also been found to be associated with heavy drinkers. This paper describes a new experimental paradigm that can be utilised to investigate attentional bias towards alcohol-related visual stimuli, specifically the ability to inhibit the orientation of initial and sustained attention, towards peripherally appearing stimuli. In this way we hope to study a novel aspect of attentional biases and how they relate to substance abuse. We used a novel eye-tracking task which aims to measure inhibitory processes for AB. The experiment utilised a gaze contingency paradigm to measure the compulsion to process or attend to alcohol stimuli. 86 undergraduate participants were recruited (31 males; 55 females), aged 18-49 years (m = 20.88; sd = 4.52). A 'break frequency' variable was computed for each participant. This was the number of times that participants tried to look at peripheral stimuli. We argue that this variable is a direct measure of how distracting peripheral stimuli were. It was found that reported alcohol use was associated with the eye-tracking break frequency measure of inhibitory control. Thus, heavy drinking may be associated with decreased inhibitory control and increased attentional bias. Results suggest that attentional bias is not just a process of stimuli becoming prioritised, but also stimuli becoming compulsory to attend and process.


Synthesis of pH-Cleavable dPG-Amines for Gene Delivery Application.
The development of effective nonviral vectors for gene therapy is still a challenge in research, due to the high toxicity of many existing polycationic nanocarriers. In this paper, the development of two pH-cleavable polyglycerol-amine-based nanocarriers is described. The benz-acetal bond represents the pH-sensitive cleavage site between dendritic polyglycerol (dPG) and glycerol-based 1,2-diamines that can complex genetic material. Due to the acid lability of the acetal moiety, the cleavable dPG-amines are less toxic in vitro. Cell-mediated degradation results in non-toxic dPG with low amine functionalization and low molecular weight cleavage products (cp). The genetic material is released because of the loss of multivalent amine groups. Interestingly, the release kinetics at the endosomal pH could be controlled by simple chemical modification of the acetals. In vitro experiments demonstrate the ability of the cleavable dPG-amine to transfect HeLa cells with GFP-DNA, which resulted in cell-compatible cleavage products.


Breastfeeding duration, costs, and benefits of a support program for low-income breastfeeding women.
Breastfeeding can ameliorate some of the complex health issues faced by low-income families. Women who breastfeed and their infants have lower health care costs compared with those who formula feed. Increasing the duration of breastfeeding is recognized as a national priority, particularly for low-income women. This community-based randomized clinical trial involving low-income mothers compared usual care with an intervention comprising hospital and home visits, and telephone support by a community health nurse/peer counselor team for 6 months after delivery. Forty-one women were recruited after delivery of a full-term singleton infant and randomly assigned to intervention or usual care groups. Women receiving the community health intervention breastfed longer than the women receiving usual care. The infants in the intervention group had fewer sick visits and reported use of fewer medications than infants in the usual care group. The intervention cost ($301/mother) was partially offset by cost savings on formula and health care. Community health nurse and peer counselor support can increase breastfeeding duration in low-income women, and has the potential to reduce total costs including the cost of support.


Mouse BMD quantitative trait loci show improved concordance with human genome-wide association loci when recalculated on a new, common mouse genetic map.
Bone mineral density (BMD) is a heritable trait, and in mice, over 100 quantitative trait loci (QTLs) have been reported, but candidate genes have been identified for only a small percentage. Persistent errors in the mouse genetic map have negatively affected QTL localization, spurring the development of a new, corrected map. In this study, QTLs for BMD were remapped in 11 archival mouse data sets using this new genetic map. Since these QTLs all were mapped in a comparable way, direct comparisons of QTLs for concordance would be valid. We then compared human genome-wide association study (GWAS) BMD loci with the mouse QTLs. We found that 26 of the 28 human GWAS loci examined were located within the confidence interval of a mouse QTL. Furthermore, 14 of the GWAS loci mapped to within 3 cM of a mouse QTL peak. Lastly, we demonstrated that these newly remapped mouse QTLs can substantiate a candidate gene for a human GWAS locus, for which the peak single-nucleotide polymorphism (SNP) fell in an intergenic region. Specifically, we suggest that MEF2C (human chromosome 5, mouse chromosome 13) should be considered a candidate gene for the genetic regulation of BMD. In conclusion, use of the new mouse genetic map has improved the localization of mouse BMD QTLs, and these remapped QTLs show high concordance with human GWAS loci. We believe that this is an opportune time for a renewed effort by the genetics community to identify the causal variants regulating BMD using a synergistic mouse-human approach.


Gray matter volume covariance networks associated with social networks in older adults.
Extensive social networks are associated with better physical, mental, and cognitive health in aging, but the underlying brain substrates remain largely unexplored. Voxel-based morphometry and multivariate statistics were used to identify gray matter volume covariance networks associated with social networks in 86 older adults without dementia (M Age = 75.20 years, 53% women). Gray matter networks associated with the number of high-contact social roles and the total number of network members were identified after adjusting for age, sex, education, global health, and total intracranial volume - and shared nodes included medial, lateral and orbital prefrontal, hippocampal, precuneus, insular, and cingulate regions. Greater expression of these gray matter networks was associated with better memory scores on the Free and Cued Selective Reminding Test. A more distributed network was associated with high-contact social roles than total number of networks members - also extending into amygdala and entorhinal cortex. Thus, high-contact social roles and total number of network members in older adults are associated with gray matter networks composed of regions previously linked to memory and affected by both healthy aging and Alzheimer disease - and high-contact social roles are more strongly associated with brain structures than the total number of network members.


The effect of infrapatellar fat pad resection on outcomes post-total knee arthroplasty: a systematic review.
The infrapatellar fat pad (IPFP) is resected in approximately 88 % of total knee arthroplasty (TKA) surgeries. The aim of this review is to investigate the impact of the IPFP resection on clinical outcomes post-TKA. A systematic search of five major databases for all relevant articles published until May, 2015 was conducted. Studies comparing the effect of IPFP resection and preservation on outcomes post-TKA were included. Each study was then assessed individually for level of evidence and risk of bias. Studies were then grouped into post-operative outcomes and given a level of evidence ranking based on the collective strength of evidence. The systematic review identified ten studies suitable for inclusion, with a total of 10,163 patients. Within these ten studies, six post-operative outcomes were identified; knee pain, vascularisation of the patella, range of motion (ROM), patella tendon length/patella infera, wound complications and patient satisfaction. Moderate evidence increased knee pain with IPFP resection post-TKA was found. Conflicting evidence was found for patella vascularisation and patellar tendon length post-TKA. Moderate evidence for no difference in ROM was found. One low quality study was found for wound complications and patient satisfaction. This systematic review is limited by the lack of level one randomised controlled trials (RCTs). There is however moderate level evidence that IPFP resection increases post-operative knee pain. Further level one RCTs are required to produce evidence-based guidelines regarding IPFP resection. Systematic Review Level of Evidence: 3.


Secular trends in opportunistic infections, cancers and mortality in patients with AIDS during the era of modern combination antiretroviral therapy.
The aim of the study was to estimate the incidence of, determine risk factors for, and investigate the consequences of opportunistic infections (OIs) and malignancies among patients with the acquired immune deficiency syndrome (AIDS) in the era of modern combination antiretroviral therapy (cART). Three enrolment periods (1998-2002, 2003-2005 and 2006-2012), corresponding to changes in predominant cART regimens, were compared among 1889 participants enrolled in a prospective cohort study, the Longitudinal Study of Ocular Complications of AIDS (LSOCA). Incidences of AIDS-related OIs and cancers were estimated. Multivariate logistic and Cox regression models were used to determine the effect of demographic and clinical characteristics on OIs and mortality. Between participants enrolled in the 1998-2002 and 2006-2012 enrolment periods, the incidence of OIs decreased from 27 per 1000 person-years (PY) to 11 per 1000 PY (P < 0.001), and mortality decreased from 41 per 1000 PY to 18 per 1000 PY (P < 0.0001), corresponding to improvements in cART regimens. Improvements in cART regimens led to a progressive decline in the incidence of OIs and mortality between 1999 and 2013 among patients with AIDS in the era of modern cART.


LOXL1 promoter haplotypes are associated with exfoliation syndrome in a U.S. Caucasian population.
LOXL1 is a major genetic risk factor for exfoliation syndrome (ES) and exfoliation glaucoma (EG). Recent evidence documenting reversal of risk alleles for the disease-associated missense variants R141L and G153D suggests that these variants are not causative and that they may be proxies for other unknown functional LOXL1 variants. The purpose of this study was to investigate the disease association of LOXL1 variants spanning the gene region, including the 5' and 3' regulatory regions, in a U.S. Caucasian case-control sample. Twenty-five LOXL1 single-nucleotide polymorphisms (SNPs), distributed throughout the gene, were genotyped in 196 Caucasian patients with ES/EG and 201 matched controls. Genotype data were analyzed for single SNP associations, SNP interactions, and haplotype associations. Promoter region haplotypes that included the risk alleles for rs12914489, a SNP located in the distal promoter region and independently associated with ES, and rs16958477, a SNP previously shown to affect gene transcription, were associated with increased disease risk (P=0.0008; odds ratio [OR], 2.34; 95% confidence interval [CI], 1.42-3.85) and with protective effects (P=2.3 × 10(-6); OR, 0.38; 95% CI, 0.25-0.57). Haplotypes containing rs12914489 and rs16958477 risk and protective alleles also significantly influenced the disease risk associated with missense alleles R141L and G153D. LOXL1 promoter haplotypes were identified that are significantly associated with ES/EG in a U.S. Caucasian population. These results suggest that promoter region SNPs can influence LOXL1 gene expression, potentially causing a reduction of enzyme activity that may predispose to disease.


Patellar Height Measurements on Radiograph and Magnetic Resonance Imaging in Patellar Instability and Control Patients.
Insall-Salvati (IS) ratio, modified IS ratio (mIS), Caton-Deschamps (CD) index, and Blackburne-Peel (BP) index are patellar height measurements commonly made on lateral radiographs (XRs) in patients with patellofemoral instability. More recently, magnetic resonance imaging (MRI) has been used as an additional imaging modality to measure patellar height. However, different imaging modalities have not been previously compared for all of these measurements. First, to compare the patellar height measurements of IS, mIS, CD, and BP on XR and MRI in first-time lateral patellar dislocation (LPD) patients and a control population and, second, to evaluate for the effect of sex and skeletal maturity. All first-time LPD patients with both MRI and XR of the knee within 6 weeks of injury from 2008 to 2012 were included in the LPD group (n = 112). Patients without patellofemoral instability who underwent both MRI and XR of the knee served as controls (n = 129). Two independent observers measured patellar height. Mean values were compared among the radiographic modalities, sex, and skeletal maturity status. Interobserver reliability was assessed using intraclass correlation coefficients (ICCs). Patellar height in the LPD group measured greater on MRI than on XR for all four patellar height ratios (IS: 0.11; mIS: 0.07; CD: 0.18; BP: 0.08). In controls, MRI also demonstrated higher readings than XR on two indices (CD: 0.17; BP: 0.16). Skeletally immature patients measured higher than skeletally mature patients in the majority of the measurements by 0.12 to 0.20 on MRI and XR; the majority of the measurements showed no sex differences. ICC was excellent for IS on both MRI (ICC = 0.830) and XR (ICC = 0.849) and for CD on XR (ICC = 0.786). Patellar height measured greater on MRI than XR by approximately 0.10 across four patellar height measurement indices. Skeletally immature patients measured higher than skeletally mature on both MRI and XR, but sex did not have an effect. This is a retrospective cohort study and its level of evidence is 3.


2017 AMERICAN ASSOCIATION OF CLINICAL ENDOCRINOLOGISTS/ENDOCRINE SOCIETY UPDATE ON TRANSGENDER MEDICINE: CASE DISCUSSIONS.
Increased numbers of transgender and gender-nonconforming people are presenting to physicians in the United States and abroad due to increased public recognition and acceptance and increased access to healthcare facilities. However, there are still gaps in medical knowledge among endocrinologists and other health care professionals. The purpose of these cases is to present several common clinical vignettes of transgender people presenting in an office setting that illustrate some of the key recommendations of the Endocrine Society's revised Endocrine Treatment of Gender Dysphoria/Gender Incongruent Persons guidelines, cosponsored by the American Association of Clinical Endocrinologists. Cases were developed based on these recently revised guidelines for gender-dysphoric and gender-nonconforming persons. Six cases are presented that illustrate the diagnosis, treatment, and long-term management of trans-gender children and adults based on the revised guidelines for the endocrine care of gender-dysphoric and gender-nonconforming persons. Several key teaching points are presented from the presentation of these cases. Endocrinologists should be familiar with the revised guidelines for gender-dysphoric and gender-nonconforming persons. Important aspects of care are the diagnosis of gender dysphoria, the timing of treatment with gender-affirming hormones, and the long-term monitoring for potential adverse outcomes. Long-term health outcome studies are needed to further help guide care in this unique population. BMI = body mass index GnRH = gonadotropin-releasing hormone HDL = high-density lipoprotein LDL = low-density lipoprotein.


Prevalence of spondyloarthritis in reference to HLA-B27 in the French population: results of the GAZEL cohort.
To estimate the prevalence of spondyloarthritis (SpA) in reference to HLA-B27 in the French population. In 1989, 20 625 employees of the French national gas and electricity company aged 35-50 years were enrolled in the GAZEL cohort. In 2010, 18 757 still active participants were screened by a questionnaire validated for the detection of SpA. Responders with available DNA were retained for further studies. Pelvic radiograph and HLA-B27 typing were performed in all the self-reported cases of SpA or psoriatic arthritis. Self-reported diagnosis was verified by a qualified rheumatologist. HLA-B27 determination was also performed in subjects without any SpA feature. The target population consisted of 6556 responders with available DNA. Their male:female ratio was 3.6 and their mean age was 65.5±3.3 years. A diagnosis of SpA was confirmed in 32 of the 72 self-reported cases, 75% of them being HLA-B27 positive. Estimated SpA prevalence adjusted for sex was 0.43% (95% CI 0.26% to 0.70%). HLA-B27 positivity rate in 2466 healthy controls was 6.9% (95% CI 5.9% to 7.9%). The relative risk of SpA in HLA-B27 positive individuals was 39 (95% CI 17 to 86). We estimated the prevalence of SpA in the French population in 2010 to 0.43%. With an estimated prevalence of 75.0% in SpA and 6.9% in healthy controls, HLA-B27 increased the disease risk 39-fold, as compared with HLA-B27 negative subjects.


High-sensitive troponin T measurements: what do we gain and what are the challenges?
Cardiac troponin (cTn) I and T are structural proteins unique to the heart. Detection of cTn in peripheral blood indicates cardiomyocyte damage. As acute myocardial infarction (AMI) is the most important cause of cardiomyocyte damage, cTns have become an integral part in the diagnosis of AMI. For this indication, cTns are superior to all other biomarkers and therefore are the preferred marker for the diagnosis of AMI. However, cTn indicates and provides an estimate of cardiomyocyte damage irrespective of its cause. The major limitation of contemporary cTn assays is that they are often not elevated during the initial hours of AMI. Recent advances in assay technology have led to more sensitive and precise cTn assays that will have a profound impact on clinical practice. High-sensitive cTn (hs-cTn) assays have two differentiating features from contemporary cTn assays: (i) detection of cTn in a majority of healthy persons and (ii) precise definition of what is 'normal' (=the 99th percentile). Recent multicentre studies have shown that hs-cTn assays improve the early diagnosis of patients with suspected AMI, particularly the early rule-out. To achieve best clinical use, cTn has to be interpreted as a quantitative variable. Rising and/or falling levels differentiate acute from chronic cardiomyocyte damage. The terms 'troponin-positive' and 'troponin negative' should therefore be avoided. 'Detectable' levels will become the norm and will have to be differentiated from 'elevated' levels. The differential diagnosis of a small amount of cardiomyocyte damage and therefore minor elevations of cTn is broad and includes acute and chronic cardiac disorders. The differential diagnosis of larger amount of injury and therefore more substantial elevations of cTn is largely restricted to AMI, myocarditis, and a rare patient with tako-tsubo cardiomyopathy.


Cloacal exstrophy: prenatal diagnosis before rupture of the cloacal membrane.
Embryologically, cloacal exstrophy is thought to result from persistence and subsequent rupture of the infraumbilical cloacal membrane during the fifth embryonic week. We report a case of cloacal exstrophy in which a prenatal diagnosis was made prior to rupture of the cloacal membrane. A routine ultrasound at 17 weeks' gestation demonstrated monoamniotic twins. One twin was normal, but the other was found to have a sacral myelomeningocele, "rocker-bottom" feet, splaying of the pubic rami, and a large cystic mass protruding from the infraumbilical anterior abdominal wall. A repeat ultrasound was performed at 22 weeks, with the same findings. At 26 weeks, further examination showed disappearance of the abdominal cyst, a small omphalocele, no demonstrable bladder, and the suggestion of prolapsed bowel inferior to the umbilical cord insertion. After delivery at 34 weeks, the abnormal twin was found to have the typical findings of cloacal exstrophy, myelomeningocele, bilateral lower limb anomalies, and extremely foreshortened small bowel. Rupture of the presumed cloacal membrane after 22 weeks in this case is inconsistent with our current understanding of the embryology of this anomaly, and should stimulate a reexamination of the current concepts. If the characteristic features are recognized, cloacal exstrophy can be diagnosed by prenatal ultrasound, permitting prenatal counseling and appropriate perinatal management.


Testing an Alternative Method for Estimating the Length of Fungal Hyphae Using Photomicrography and Image Processing.
This study aimed to develop and test an unbiased and rapid methodology to estimate the length of external arbuscular mycorrhizal fungal (AMF) hyphae in soil. The traditional visual gridline intersection (VGI) method, which consists in a direct visual examination of the intersections of hyphae with gridlines on a microscope eyepiece after aqueous extraction, membrane-filtration, and staining (e.g., with trypan blue), was refined. For this, (i) images of the stained hyphae were taken by using a digital photomicrography technique to avoid the use of the microscope and the method was referred to as "digital gridline intersection" (DGI) method; and (ii), the images taken in (i) were processed and the hyphal length was measured by using ImageJ software, referred to as the "photomicrography-ImageJ processing" (PIP) method. The DGI and PIP methods were tested using known grade lengths of possum fur. Then they were applied to measure the hyphal lengths in soils with contrasting phosphorus (P) fertility status. Linear regressions were obtained between the known lengths (Lknown) of possum fur and the values determined by using either the DGI (LDGI) (LDGI = 0.37 + 0.97 × Lknown, r2 = 0.86) or PIP (LPIP) methods (LPIP = 0.33 + 1.01 × Lknown, r2 = 0.98). There were no significant (P > 0.05) differences between the LDGI and LPIP values. While both methods provided accurate estimation (slope of regression being 1.0), the PIP method was more precise, as reflected by a higher value of r2 and lower coefficients of variation. The average hyphal lengths (6.5-19.4 m g-1) obtained by the use of these methods were in the range of those typically reported in the literature (3-30 m g-1). Roots growing in P-deficient soil developed 2.5 times as many hyphae as roots growing in P-rich soil (17.4 vs 7.2 m g-1). These tests confirmed that the use of digital photomicrography in conjunction with either the grid-line intersection principle or image processing is a suitable method for the measurement of AMF hyphal lengths in soils for comparative investigations.


Physical activity, weight control, and breast cancer risk and survival: clinical trial rationale and design considerations.
Substantial observational epidemiological evidence exists that physical activity and weight control are associated with decreased risk of postmenopausal breast cancer. Uncertainty remains regarding several aspects of these associations, including the effect of possible confounding factors on these associations. We present the rationale and design for two randomized controlled trials that can help resolve this uncertainty. In a 5-year prevention trial conducted among women at high risk of breast cancer, the primary endpoint would be breast cancer incidence. For a comparable survivorship trial, the primary endpoint would be the disease-free interval and secondary endpoints would be breast cancer recurrence-free interval, second primary breast cancer, and total invasive plus in situ breast cancer. A set of inclusion and exclusion criteria is proposed for both trials. Intervention goals are the same for both trials. Goals for the weight control intervention would be, for women whose body mass index (BMI) is greater than 25 kg/m(2), to lose 10% of body weight and, for women whose BMI is less than or equal to 25 kg/m(2), to avoid weight gain. The goal for the physical activity intervention would be to achieve and maintain regular participation in a moderate-intensity physical activity program for a total of 150-225 minutes over at least 5 days per week. Sample size calculations are based on alternative assumptions about hazard ratio, adherence, follow-up duration, and power and are presented for the primary prevention and survivorship trials. Although both studies could enhance our understanding of breast cancer etiology and benefit public health, practical considerations, including smaller sample size, ease of recruitment, and reduced likelihood of early termination, favor the survivorship trial at this time.


Japan Society of Gynecologic Oncology 2018 guidelines for treatment of uterine body neoplasms.
The Fourth Edition of the Guidelines for Treatment of Uterine Body Neoplasm was published in 2018. These guidelines include 9 chapters: 1. Overview of the guidelines, 2. Initial treatment for endometrial cancer, 3. Postoperative adjuvant therapy for endometrial cancer, 4. Post-treatment surveillance for endometrial cancer, 5. Treatment for advanced or recurrent endometrial cancer, 6. Fertility-sparing therapy, 7. Treatment of uterine carcinosarcoma and uterine sarcoma, 8. Treatment of trophoblastic disease, 9. Document collection; and nine algorithms: 1-3. Initial treatment of endometrial cancer, 4. Postoperative adjuvant treatment for endometrial cancer, 5. Treatment of recurrent endometrial cancer, 6. Fertility-sparing therapy, 7. Treatment for uterine carcinosarcoma, 8. Treatment for uterine sarcoma, 9. Treatment for choriocarcinoma. Each chapter includes overviews and clinical questions, and recommendations, objectives, explanation, and references are provided for each clinical question. This revision has no major changes compared to the 3rd edition, but does have some differences: 1) an explanation of the recommendation decision process and conflict of interest considerations have been added in the overview, 2) nurses, pharmacists and patients participated in creation of the guidelines, in addition to physicians, 3) the approach to evidence collection is listed at the end of the guidelines, and 4) for clinical questions that lack evidence or clinical validation, the opinion of the Guidelines Committee is given as a "Recommendations for tomorrow".


Violence as a public health problem: an ecological study of 169 countries.
Individual level risk factors for violence have been widely studied, but little is known about country-level determinants, particularly in low and middle-income countries. We hypothesized that income inequality, through its detrimental effects on social cohesion, would be related to an increase in violence worldwide, and in low and middle-income countries in particular. We examined country-level associations of violence with socio-economic and health-related factors, using crime statistics from the United Nations Office on Drugs and Crime, and indicators from the Human Development Report published by the United Nations Development Programme. Using regression models, we measured relationships between country-level factors (age, education, measures of income, health expenditure, and alcohol consumption) and four violent outcomes (including measures of violence-related mortality and morbidity) in up to 169 countries. We stratified our analyses comparing high with low and middle-income countries, and analysed longitudinal data on homicide and income inequality in high-income countries. In low and middle-income countries, income inequality was related to homicide, robbery, and self-reported assault (all p's < 0.05). In high-income countries, urbanicity was significantly associated with official assault (p = 0.002, β = 0.716) and robbery (p = 0.011, β = 0.587) rates; income inequality was related to homicide (p = 0.006, β = 0.670) and self-reported assault (p = 0.020, β = 0.563), and longitudinally with homicide (p = 0.021). Worldwide, alcohol consumption was associated with self-reported assault rates (p < 0.001, β = 0.369) suggesting public policy interventions reducing alcohol consumption may contribute to reducing violence rates. Our main finding was that income inequality was related to violence in low and middle-income countries. Public health should advocate for global action to moderate income inequality to reduce the global health burden of violence.


Second and subsequent tumours among 1927 retinoblastoma patients diagnosed in Britain 1951-2004.
Retinoblastoma is an eye tumour of childhood that occurs in heritable and non-heritable forms. In the heritable form, there is a predisposition to the development of non-ocular subsequent primary tumours (SPTs). This study included 1927 retinoblastoma patients diagnosed in Britain from 1951 to 2004. Ascertainment was through the (UK) National Registry of Childhood Tumours; cases were followed-up for the occurrence of SPTs. Standardised incidence ratios (SIRs) were calculated. We identified 169 SPTs in 152 patients. The SIR analysis included 145 SPTs with cancer registrations from the years 1971 to 2009. These tumours occurred in 132 patients: 112 of the 781 heritable and 20 of the 1075 (presumed) non-heritable cases under surveillance at the start of this period developed at least one registered SPT. The SIRs for all tumours combined were 13.7 (95% confidence interval 11.3-16.5) in heritable cases and 1.5 (0.9-2.3) in non-heritable cases. The main types of SPT in the heritable cases were leiomyosarcoma, (31 cases; SIR 1018.7 (692.2-1446.0)), osteosarcoma (26 cases; SIR 444.6 (290.4-651.4)), and skin melanoma (12 cases; SIR 18.6 (9.6-32.4)). The risk of SPTs in heritable retinoblastoma is extremely high. This has important implications for the clinical follow-up and counselling of survivors and their families.


Pediatric arytenoid dislocation: diagnosis and treatment.
Arytenoid dislocation and subluxation are well-described injuries in adults but are poorly documented in children. The most commonly cited etiology is intubation trauma although external blunt trauma also is recognized. Symptoms include dysphonia, vocal fatigue, loss of vocal control, breathiness, odynophagia, dysphagia, dyspnea, and cough. Prompt diagnosis and treatment lead to the best chance for recovery, and delayed treatment is likely to result in scarring and possibly ankylosis. The mean age of our study group was 12.3 years and consisted of six males (55%) and five females (45%). The most commonly presenting symptom was hoarseness (81.8%). Six of the 11 patients underwent surgical correction of the dislocated arytenoid cartilage. Four patients refused any treatment and one patient received voice therapy alone. Two patients who refused surgical intervention had spontaneous reduction of their dislocations. After surgical intervention, one patient regained normal voice, four patients had substantial voice improvement without return to preinjury vocal function, and one patient had only slight voice improvement. Pediatric symptoms are similar to these in adults, yet these may be less noticeable to the patient and clinician. A high index of suspicion is needed to diagnose and treat pediatric arytenoid dislocation.


Risk of herpes zoster in CKD: a matched-cohort study based on administrative data.
Immune system dysregulation is associated with end-stage renal disease. Although decreased cellular immunity increases susceptibility to herpes zoster, the risk of herpes zoster in patients with earlier stages of chronic kidney disease (CKD) is unclear. A matched-cohort study. Data from the Taiwan Longitudinal Health Insurance Database (LHID) for 2004-2006 were analyzed. The study cohort included patients 18 years or older given a diagnosis of CKD (excluding patients treated by dialysis or transplant) in 2004-2005 (n = 13,321). The comparison cohort (n = 66,605) included 5 randomly selected age- and sex-matched controls for each patient in the study cohort. CKD. Incident cases of CKD were identified using the Taiwan LHID. CKD was ascertained from International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) codes. Herpes zoster, ascertained from ICD-9-CM codes. All participants were followed up from the date of cohort entry until they developed herpes zoster or the end of 2006. Cox proportional hazard regressions were performed to compare the hazard rates of herpes zoster in the CKD cohort and the age- and sex-matched comparison cohort. We identified 13,321 patients with a diagnosis of CKD who matched the inclusion criteria. 1,602 patients developed herpes zoster during the study period, of whom 353 were from the CKD cohort and 1,249 were from the comparison cohort. After adjusting for potential confounding factors, CKD was associated independently with greater risk of herpes zoster (HR, 1.60; 95% CI, 1.41-1.81). Some patients with CKD or herpes zoster may have chosen not to seek medical care. Misclassification of CKD due to use of diagnostic codes also is a limitation. This population-based cohort study indicated that patients with CKD are at increased risk of herpes zoster compared with the general population.


Interaction of Cryptococcus neoformans Rim101 and protein kinase A regulates capsule.
Cryptococcus neoformans is a prevalent human fungal pathogen that must survive within various tissues in order to establish a human infection. We have identified the C. neoformans Rim101 transcription factor, a highly conserved pH-response regulator in many fungal species. The rim101 multiply sign in circle mutant strain displays growth defects similar to other fungal species in the presence of alkaline pH, increased salt concentrations, and iron limitation. However, the rim101 multiply sign in circle strain is also characterized by a striking defect in capsule, an important virulence-associated phenotype. This capsular defect is likely due to alterations in polysaccharide attachment to the cell surface, not in polysaccharide biosynthesis. In contrast to many other C. neoformans capsule-defective strains, the rim101 multiply sign in circle mutant is hypervirulent in animal models of cryptococcosis. Whereas Rim101 activation in other fungal species occurs through the conserved Rim pathway, we demonstrate that C. neoformans Rim101 is also activated by the cAMP/PKA pathway. We report here that C. neoformans uses PKA and the Rim pathway to regulate the localization, activation, and processing of the Rim101 transcription factor. We also demonstrate specific host-relevant activating conditions for Rim101 cleavage, showing that C. neoformans has co-opted conserved signaling pathways to respond to the specific niche within the infected host. These results establish a novel mechanism for Rim101 activation and the integration of two conserved signaling cascades in response to host environmental conditions.


Gait detection in children with and without hemiplegia using single-axis wearable gyroscopes.
In this work, we develop a novel gait phase detection algorithm based on a hidden Markov model, which uses data from foot-mounted single-axis gyroscopes as input. We explore whether the proposed gait detection algorithm can generate equivalent results as a reference signal provided by force sensitive resistors (FSRs) for typically developing children (TD) and children with hemiplegia (HC). We find that the algorithm faithfully reproduces reference results in terms of high values of sensitivity and specificity with respect to FSR signals. In addition, the algorithm distinguishes between TD and HC and is able to assess the level of gait ability in patients. Finally, we show that the algorithm can be adapted to enable real-time processing with high accuracy. Due to the small, inexpensive nature of gyroscopes utilized in this study and the ease of implementation of the developed algorithm, this work finds application in the on-going development of active orthoses designed for therapy and locomotion in children with gait pathologies.


The type III TGFβ receptor regulates filopodia formation via a Cdc42-mediated IRSp53-N-WASP interaction in epithelial cells.
Cell adhesion and migration are tightly controlled by regulated changes in the actin cytoskeleton. Previously we reported that the TGFβ (transforming growth factor β) superfamily co-receptor, TβRIII (type III TGFβ receptor; also known as βglycan), regulates cell adhesion, migration and invasion, and suppresses cancer progression, in part, through activation of the small GTPase Cdc42 (cell division cycle 42), and Cdc42-dependent alterations to the actin cytoskeleton. In the present study we demonstrate that TβRIII specifically promotes filopodial formation and extension in MCF10A and HMEC (human mammary epithelial cell) mammary epithelial cells. Mechanistically, cell-surface TβRIII and Cdc42 co-localize to filopodial structures and co-complex in a β-arrestin2-dependent, and a TβRI/TβRII-independent manner. The β-arrestin2-mediated interaction between TβRIII and Cdc42 increases complex formation between the Cdc42 effectors IRSp53 with N-WASP (neuronal Wiskott-Aldrich syndrome protein) to increase filopodial formation. We demonstrate a function link between filopodial structures and epithelial cell adhesion as regulated by the TβRIII-Cdc42 interaction. The present studies identify TβRIII as a novel regulator of IRSp53/N-WASP via Cdc42 to regulate filopodial formation and cell adhesion.


Nongenomic actions of bile acids. Synthesis and preliminary characterization of 23- and 6,23-alkyl-substituted bile acid derivatives as selective modulators for the G-protein coupled receptor TGR5.
23-Alkyl-substituted and 6,23-alkyl-disubstituted derivatives of chenodeoxycholic acid are identified as potent and selective agonists of TGR5, a G-protein coupled receptor for bile acids (BAs). In particular, we show that methylation at the C-23(S) position of natural BAs confers a marked selectivity for TGR5 over FXR, while the 6alpha-alkyl substitution increases the potency at both receptors. The present results allow for the first time a pharmacological differentiation of genomic versus nongenomic effects mediated by BA derivatives.


Impaired Mitochondrial Microbicidal Responses in Chronic Obstructive Pulmonary Disease Macrophages.
Chronic obstructive pulmonary disease (COPD) is characterized by impaired clearance of pulmonary bacteria. The effect of COPD on alveolar macrophage (AM) microbicidal responses was investigated. AMs were obtained from bronchoalveolar lavage from healthy donors or patients with COPD and challenged with opsonized serotype 14 Streptococcus pneumoniae. Cells were assessed for apoptosis, bactericidal activity, and mitochondrial reactive oxygen species (mROS) production. A transgenic mouse line in which the CD68 promoter ensures macrophage-specific expression of human induced myeloid leukemia cell differentiation protein Mcl-1 (CD68.hMcl-1) was used to model the molecular aspects of COPD. COPD AMs had elevated levels of Mcl-1, an antiapoptotic B-cell lymphoma 2 family member, with selective reduction of delayed intracellular bacterial killing. CD68.hMcl-1 AMs phenocopied the microbicidal defect because transgenic mice demonstrated impaired clearance of pulmonary bacteria and increased neutrophilic inflammation. Murine bone marrow-derived macrophages and human monocyte-derived macrophages generated mROS in response to pneumococci, which colocalized with bacteria and phagolysosomes to enhance bacterial killing. The Mcl-1 transgene increased oxygen consumption rates and mROS expression in mock-infected bone marrow-derived macrophages but reduced caspase-dependent mROS production after pneumococcal challenge. COPD AMs also increased basal mROS expression, but they failed to increase production after pneumococcal challenge, in keeping with reduced intracellular bacterial killing. The defect in COPD AM intracellular killing was associated with a reduced ratio of mROS/superoxide dismutase 2. Up-regulation of Mcl-1 and chronic adaption to oxidative stress alter mitochondrial metabolism and microbicidal function, reducing the delayed phase of intracellular bacterial clearance in COPD.


[Inhibitory effects of saponins from Tribulus terrestris on alpha-glucosidase in small intestines of rats].
To determine the effects of saponins from Tribulus terrestris (STT) on small intestinal a-glucosidase and postprandial blood glucose levels in rats. The inhibitory effects of STT on a-glucosidase extracted from small intestines in rats were carried out in vitro. The blood glucose levels were measured after 60 min when sucrose (2 g x kg(-1)) or glucose (2 g x kg(-1)) was administered orally with STT (100 mg x kg(-1)). After treated with STT (100 mg x kg(-1)) for 14 d, the activities of a-glucosidase were determined daily, as well as the postprandial blood glucose levels after oraly administered sucrose (2 g x kg(-1)). STT at concentrations of 0.1, 1 and 10 mg x mL(-1) reduced significantly the activities of alpha-glucosidase with inhibitory rates of (20.83 +/- 1.66)%, (43.73 +/- 2.39)% and (52.62 +/- 2.69)%, respectively. In facts STT (100 mg x kg(-1)) considerably decreased the blood glucose levels which was 52.61% of that of the control in rats co-administered orally with sucrose (2 g x kg(-1)). However, it showed no such effect on the rats co-administered orally with glucose (2 g x kg(-1)). After orally administered of STT for 14 d, the activity of alpha-glucosidase was significantly reduced (P < 0.05) to (58.17 +/- 3.24)% of that those in control. Meanwhile, The rats were oral administered with sucrose, the increase of postprandial blood glucose levels were (69.50 +/- 4.28)% of that in control 60 min later ( P < 0.05). It was through inhibiting the activity of a-glucosidase in small intestines that STT significantly retarded the increase in postprandial blood glucose levels in rats.


Altered Perception of Facially Expressed Tiredness in Insomnia.
The present study compared normal sleepers and individuals displaying insomnia symptoms in their ratings for the expression intensity of tiredness and alertness whilst observing tired and neutral faces. Fifty-six normal sleepers and 58 individuals with insomnia symptoms observed 98 facial photographs (49 neutral, 49 tired). Using a visual analogue scale, participants were required to rate the extent to which each face appeared as tired and alert. Tired faces were created by manipulating neutral photographs to include previously identified facial tiredness cues. All participants rated sleep-related faces as more tired and less alert relative to neutral photographs. A significant Group × Face × Rating interaction demonstrated that, compared with normal sleepers, the insomnia symptoms group showed lower ratings for the expression of tiredness, but not alertness, whilst observing the tired faces. The findings suggest that the presence of insomnia symptoms is associated with reduced ratings of expression intensity for sleep-related facial photographs displaying tiredness. These outcomes add to the body of literature on how facial cues of tiredness are perceived by those with insomnia symptoms. Further work is required to elucidate the mechanisms underlying the relationship between insomnia symptoms and reduced perceptions of facially expressed tiredness.


Quality and cost analysis of nurse staffing, discharge preparation, and postdischarge utilization.
To determine the impact of unit-level nurse staffing on quality of discharge teaching, patient perception of discharge readiness, and postdischarge readmission and emergency department (ED) visits, and cost-benefit of adjustments to unit nurse staffing. Patient questionnaires, electronic medical records, and administrative data for 1,892 medical-surgical patients from 16 nursing units within four acute care hospitals between January and July 2008. Nested panel data with hospital and unit-level fixed effects and patient and unit-level control variables. Registered nurse (RN) staffing was recorded monthly in hours-per-patient-day. Patient questionnaires were completed before discharge. Thirty-day readmission and ED use with reimbursement data were obtained by cross-hospital electronic searches. Higher RN nonovertime staffing decreased odds of readmission (OR=0.56); higher RN overtime staffing increased odds of ED visit (OR=1.70). RN nonovertime staffing reduced ED visits indirectly, via a sequential path through discharge teaching quality and discharge readiness. Cost analysis projected total savings from 1 SD increase in RN nonovertime staffing and decrease in RN overtime of U.S.$11.64 million and U.S.$544,000 annually for the 16 study units. Postdischarge utilization costs could potentially be reduced by investment in nursing care hours to better prepare patients before hospital discharge.


Continuing professional development (CPD) in radiography: A collaborative European meta-ethnography literature review.
The aim of the study was to complete a collaborative review of Radiography continuing professional development (CPD) research material to support the production of European Federation of Radiographer Societies (EFRS) CPD recommendations. A meta-ethnography approach to literature review was applied focussing upon commonalities rather than discrepancies between research outcomes. This facilitated exploration of context across the geographical region of Europe with national variations in CPD governance. The seven phases of the meta-ethnographic approach were followed by two independent experienced researchers. A third researcher mediated the findings which were then explored collaboratively with the EFRS CPD working group for concordance. Phase seven of the meta-ethnography involved interpreting an expression of the synthesis from the previous stages. Six main corroborating themes emerged in this process and following mediation were expressed as themes; knowledge, skills & competency, needs/gap analysis, multi-layered/multi-modal, barriers and drivers; regulation vs autonomy; fostering collaboration - harnessing technology. The primary feature of CPD activity should be the resulting impact - to patients, the service, the profession and the individual; with all stakeholders working in partnership. CPD activity must be flexible/multi-modal to support the changing growth/dynamic workforce. All stakeholders should utilise communication and technology resources and make efforts to improve collaboration between the management, regulators and educators to support Radiographers to develop meaningful CPD. Health services across Europe are under increasing stress and a principal factor going forwards will be managing increasing demands on healthcare staff whilst supporting enhancement of the knowledge, skills and competency base.


Comparison of cannabinoids with known analgesics using a novel high throughput zebrafish larval model of nociception.
It has been established that both adult and larval zebrafish are capable of showing nociceptive responses to noxious stimuli; however, the use of larvae to test novel analgesics has not been fully explored. Zebrafish larvae represent a low-cost, high-throughput alternative to traditional mammalian models for the assessment of product efficacy during the initial stages of drug development. In the current study, a novel model of nociception using zebrafish larvae is described. During the recovery from an acute exposure to low levels of acetic acid, larvae display innate changes in behaviour that may be indicative of nociception. To assess the usefulness of this model for testing potential analgesics, three known synthetic pain medications were assessed (ibuprofen, acetaminophen and tramadol) along with three naturally occurring products (honokiol, tetrahydrocannabinol and cannabidiol). When the effect of each compound on both the acetic acid recovery and control activity was compared there appeared to be both similarities and differences between the compounds. One of the most interesting effects was found for cannabidiol which appeared to oppose the activity change during the recovery period of AA exposed larvae while having a nominal effect on control activity. This would appear to be in line with current research that has demonstrated the nociceptive properties of cannabidiol. Here we have provided a novel model that will complement existing zebrafish models and will expand on the potential use of zebrafish larvae for studying both nociception and new analgesics.


Non-Hodgkin's lymphoma of the head and neck: CT evaluation of nodal and extranodal sites.
Forty-five patients with non-Hodgkin's lymphoma (NHL) of the extracranial head and neck who had undergone CT as part of their evaluation were reviewed to assess the impact of CT on clinical management. The sites of tumor deposition were subdivided by location: I, nodal; II, extranodal, lymphatic (Waldeyer's ring); and III, extranodal, extralymphatic (orbit, sinonasal, deep facial spaces, mandible, salivary gland, skin, and larynx). The CT appearance of NHL in each of the three locations was analyzed for characteristic CT signatures. Nodal NHL was suspected when CT showed multiple, large, homogeneous lymph nodes, often in unusual nodal chains of the head and neck. Extranodal, lymphatic NHL of Waldeyer's ring was indistinguishable from squamous cell carcinoma of this area unless synchronous tumor deposit in an extranodal, extralymphatic location was also present. When NHL was in nodes and/or Waldeyer's ring, CT-derived information was of limited clinical value since treatment was unfocused (chemotherapy and/or large-field radiotherapy). The CT appearances of extranodal, extralymphatic NHL was generally not distinguishable from other malignancies of these areas. However, CT-derived information regarding deep-tissue tumor size and extent was critical to planning the radiotherapy ports.


Attenuation of low-frequency sound in freshwater.
Lake Superior was chosen as an experimental site to compare sound absorption of freshwater with the results of absorption measurements in seawater. The relaxation-like absorption at 1 kilohertz occurring in seawater is also present in freshwater. A relaxation related to the structural characteristics rather than to the salt content of water may be responsible for the anomalous absorption.


Modeling the genealogy of a cultural trait.
The mathematical study of genealogies has yielded important insights in population biology, such as the ability to estimate the time to the most recent common ancestor (MRCA) of a sample of genetic sequences or of a group of individuals. Here we introduce a model of cultural genealogies that is a step toward answering similar questions for cultural traits. In our model individuals can inherit from a variable, potentially large number of ancestors, rather than from a fixed, small number of ancestors (one or two) as is typical of genetic evolution. We first show that, given a sample of individuals, a cultural common ancestor does not necessarily exist. We then introduce a related concept: the most recent unique ancestor (MRUA), i.e., the most recent single individual who is the earliest cultural ancestor of the sample. We show that, under neutral evolution, the time to the MRUA can be staggeringly larger than the time to MRCA in a single ancestor model, except when the average number of learning opportunities per individuals is small. Our results point out that the properties of cultural genealogies may be very different from those of genetic genealogies, with potential implications for reconstructing the histories of cultural traits.


Prenatal and early postnatal measures of brain development and childhood sleep patterns.
BackgroundBrain development underlies maturation of sleep patterns throughout childhood. Intrauterine head growth-marker of early neurodevelopment-has not been associated with childhood sleep characteristics. We explored associations between ultrasonographic measures of prenatal and early postnatal neurodevelopment and childhood sleep.MethodsA total of 6,808 children from a population-based birth cohort (Generation R) were included. Head circumference (HC) and lateral ventricles size were assessed with mid- and late-pregnancy fetal ultrasounds, and with cranial ultrasound 3-20 weeks postnatally. Mothers reported children's sleep duration at 2 and 3 years, and sleep problems at 1.5, 3, and 6 years.ResultsLarger ventricular size, but not HC, was related to longer sleep duration at 3 years (β=0.06 h, 95% confidence interval (CI): 0.02; 0.10 in late-pregnancy and β=0.11 h, 95% CI: 0.02; 0.20 in early infancy, mid-pregnancy parameters were unrelated to sleep duration). Larger HC in mid-pregnancy was associated with a reduced risk for being a "problematic sleeper" up to the age of 6 years (odds ratio (OR): 0.94, 95% CI: 0.89; 0.99). Consistently, children with larger HC in early infancy were less likely to be "problematic sleepers" at 3 and 6 years.ConclusionsThis study shows that variations in fetal and neonatal brain size may underlie behavioral expression of sleep in childhood. Albeit small effect estimates, these associations provide evidence for neurodevelopmental origins of sleep.


Synthesis, Biological Evaluation and Molecular Modeling of Substituted Indeno[1,2-b]indoles as Inhibitors of Human Protein Kinase CK2.
Due to their system of annulated 6-5-5-6-membered rings, indenoindoles have sparked great interest for the design of ATP-competitive inhibitors of human CK2. In the present study, we prepared twenty-one indeno[1,2-b]indole derivatives, all of which were tested in vitro on human CK2. The indenoindolones 5a and 5b inhibited human CK2 with an IC50 of 0.17 and 0.61 µM, respectively. The indeno[1,2-b]indoloquinone 7a also showed inhibitory activity on CK2 at a submicromolar range (IC50 = 0.43 µM). Additionally, a large number of indenoindole derivatives was evaluated for their cytotoxic activities against the cell lines 3T3, WI-38, HEK293T and MEF.


Arterial smooth muscle cells express nitric oxide synthase in response to endothelial injury.
Endothelial cells regulate vascular tone by secreting paracrine mediators that control the contractility of arterial smooth muscle cells. Nitric oxide (NO) is an important vasodilating agent that is generated from L-arginine by the enzyme nitric oxide synthase (NOS), which is expressed constitutively by the endothelium. NO also inhibits platelet aggregation, contributing to the antithrombotic properties of the endothelial surface. It would therefore be expected that loss of the endothelium during arterial injury would lead to vasospasm and thrombosis but instead, the neointima formed after injury has a nonthrombogenic surface and a maintained vascular patency. We report here that arterial smooth muscle cells in the neointima formed after a deendothelializing balloon injury to the rat carotid artery express the cytokine-inducible isoform of NOS. Expression was detectable by reverse transcription-polymerase chain reaction from day 1-14 after injury and in situ hybridization showed expression of NOS mRNA by neointimal smooth muscle cells, particularly at the surface of the lesion. This was associated with systemically detectable NO production as revealed by electron paramagnetic resonance spectroscopic analysis of nitrosylated red cell hemoglobin. Local NO production by intimal smooth muscle cells after endothelial injury could represent an important mechanism for the maintenance of arterial patency and nonthrombogenicity in the injured artery.