Use of the comet-FISH assay to demonstrate repair of the TP53 gene region in two human bladder carcinoma cell lines.
The alkaline single-cell gel electrophoresis (comet) assay can be combined with fluorescence in situ hybridization (FISH) methodology to investigate the localization of specific gene domains within an individual cell. The position of the fluorescent hybridization spots in the comet head or tail indicates whether the sequence of interest lies within or in the vicinity of a damaged region of DNA. In this study, we used the comet-FISH assay to examine initial DNA damage and subsequent repair in the TP53 gene region of RT4 and RT112 bladder carcinoma cells after 5 Gy gamma irradiation. In addition to standard comet parameter measurements, the number and location of TP53 hybridization spots within each comet was recorded at each repair time. The results indicate that the rate of repair of the TP53 gene region was fastest during the first 15 min after damage in both cell lines. When compared to overall genomic repair, the repair of the TP53 gene region was observed to be significantly faster during the first 15 min and thereafter followed a rate similar to that for the overall genome. The data indicate that the TP53 domain in RT4 and RT112 cells is repaired rapidly after gamma irradiation. Furthermore, this repair may be preferential compared to the repair of overall genomic DNA, which gives a measure of the average DNA repair response of the whole genome. We suggest that the comet-FISH assay has considerable potential in the study of gene-specific repair after DNA damage.


Angelman syndrome phenotype associated with mutations in MECP2, a gene encoding a methyl CpG binding protein.
Angelman syndrome (AS) is a neurodevelopmental disorder characterised by severe mental retardation, absent speech, ataxia, sociable affect, and dysmorphic facial features. Eighty five percent of patients with AS have an identifiable genetic abnormality of chromosome 15q11-13. Mutations within the X linked MECP2 gene have been identified in patients with Rett syndrome (RTT), a neurodevelopmental disorder which affects females almost exclusively and which shares phenotypic overlap with AS. RTT is usually associated with normal development in infancy followed by loss of acquired skills and evolution of characteristic hand wringing movements and episodes of hyperventilation.A panel of 25 female and 22 male patients with a clinical diagnosis of AS and no molecular abnormality of 15q11-13 were screened for MECP2 mutations and these were identified in four females and one male. Following the diagnosis, it was possible to elicit a history of regression in three of these patients, who by then were showing features suggestive of Rett syndrome. In the remaining two subjects the clinical phenotype was still considered to be Angelman-like. These findings illustrate the phenotypic overlap between the two conditions and suggest that screening for MECP2 mutations should be considered in AS patients without a demonstrable molecular or cytogenetic abnormality of 15q11-13. Since MECP2 mutations almost always occur de novo, their identification will substantially affect genetic counselling for the families concerned.


Tomographic motion detection and correction directly in sinogram space.
Patient motion, especially respiratory motion, results in various artefacts such as blurring and streaks in tomographic images. The interplay of the movement of the beam aperture and variations of organ anatomy during delivery can create 'hot' and 'cold' spots throughout the field in intensity-modulated radiation therapy (IMRT). Detection and correction of patient motion is extremely important in tomographic imaging and IMRT. Tomographic projection data (sinogram) encode not only the patient anatomy information, but also the intra-scanning motion information. In this paper, we developed an algorithm to detect and correct the in-plane respiratory motion directly in sinogram space. The respiratory motion is modelled as time-varying scaling along the x and y directions. Its effects on the sinogram are discussed. Based on the traces of some nodal points in the sinogram, the intra-scanning motion is determined. The motion correction is also implemented in sinogram space. The motion-corrected sinogram is used for reconstruction by the filtered back-projection (FBP) method. Computer simulations validate the motion detection and correction algorithm. The reconstructed images from the motion-corrected sinogram eliminate the majority of the artefacts. The method could be applied to projection data used in CT and ECT, as well as in tomotherapy delivery modification and dose reconstruction.


Comparative Outcomes in Functional Rhinoplasty With Open vs Endonasal Spreader Graft Placement.
Spreader grafts are commonly used for nasal valve stenosis. There is debate among practitioners regarding the optimal approach for spreader graft placement. This study aimed to determine whether an endonasal spreader graft placement leads to equivalent postoperative outcomes using a standardized nasal obstruction symptom score and whether there is a significant difference in operative time between the 2 approaches. A retrospective review of functional rhinoplasties by a single facial plastic surgeon over a 2-year period was conducted. Facial plastic surgery practice at a tertiary academic medical center. Fifty patients with a single surgeon over a 22-month period with a minimum of 1-year follow-up were included in the study. Age, sex, and preoperative Nasal Obstruction Symptom Evaluation (NOSE) scores of patients were equivalent between the 2 groups. For time analysis, 56 of 107 patients were analyzed, with exclusion of patients undergoing combined procedures and autologous cartilage grafting from sites besides the nasal septum. Patients who underwent functional rhinoplasty through either an open or an endonasal (closed) approach were compared. Preoperative approach decision was based on the need for cosmetic tip, dorsal, or anterior septal work, and all decisions were made before the development of this study. Main outcome measures were as follows: improvement in NOSE scores recorded preoperatively and at 1-year minimum postoperative follow-up as well as difference in operative times. No significant difference (P = 0.92) was found between patients having open or endonasal spreader graft placement in NOSE score improvement. Open rhinoplasty was associated with significantly longer operative times (P < 0.001), and performance of additional maneuvers such as strut grafts, osteotomies, and dorsal hump reduction was not found to independently affect operative times significantly. Spreader grafts can be placed through an endonasal or open approach with similar outcomes in a standardized measure. Open rhinoplasty is associated with prolonged operative times and therefore increased operative costs. In properly selected patients, endonasal spreader graft placement may lead to significant cost savings when open rhinoplasty is not otherwise indicated.


Functional outcome in contemporary children and young adults with tetralogy of Fallot after repair.
Functional outcome measures are of growing importance in the aftercare of patients with congenital heart disease. This study addresses the functional status with regard to exercise capacity, health-related physical fitness (HRPF) and arterial stiffness in a recent cohort of children, adolescents and young adults with tetralogy of Fallot (ToF) after repair. Single-centre, uncontrolled and prospective cohort study. Outpatient department of the German Heart Centre Munich; July 2014-January 2018. One hundred and six patients with ToF after repair (13.5±3.7 years, 40 females) were included. Data were compared with a recent cohort of healthy controls (HCs) (n=1700, 12.8±2.6 years, 833 females). Patients underwent a symptom-limited cardiopulmonary exercise test, performed an HRPF test (FitnessGram) and had an assessment of their arterial stiffness (Mobil-O-Graph). Compared with HC, patients with ToF showed lower predicted [Formula: see text]O2 peak (ToF: 80.4% ± 16.8% vs HC: 102.6% ± 18.1%, p<0.001), impaired ventilatory efficiency (ToF: 29.6 ± 3.6 vs HC: 27.4 ± 2.9, p<0.001), chronotropic incompetence (ToF: 167 ± 17 bpm vs HC: 190 ± 17 bpm, p<0.001) and reduced HRPF (ToF z-score: -0.65 ± 0.87 vs HC z-score: 0.03 ± 0.65, p<0.001). Surrogates of arterial stiffness, central and peripheral systolic blood pressure, did not differ between the two groups. Contemporary children, adolescents and young adults with ToF still have functional limitations. How impaired HRPF and limited exercise capacity interact and how they can be modified needs to be evaluated in further intervention studies.


The Neural Time Course of Semantic Ambiguity Resolution in Speech Comprehension.
Semantically ambiguous words challenge speech comprehension, particularly when listeners must select a less frequent (subordinate) meaning at disambiguation. Using combined magnetoencephalography (MEG) and EEG, we measured neural responses associated with distinct cognitive operations during semantic ambiguity resolution in spoken sentences: (i) initial activation and selection of meanings in response to an ambiguous word and (ii) sentence reinterpretation in response to subsequent disambiguation to a subordinate meaning. Ambiguous words elicited an increased neural response approximately 400-800 msec after their acoustic offset compared with unambiguous control words in left frontotemporal MEG sensors, corresponding to sources in bilateral frontotemporal brain regions. This response may reflect increased demands on processes by which multiple alternative meanings are activated and maintained until later selection. Disambiguating words heard after an ambiguous word were associated with marginally increased neural activity over bilateral temporal MEG sensors and a central cluster of EEG electrodes, which localized to similar bilateral frontal and left temporal regions. This later neural response may reflect effortful semantic integration or elicitation of prediction errors that guide reinterpretation of previously selected word meanings. Across participants, the amplitude of the ambiguity response showed a marginal positive correlation with comprehension scores, suggesting that sentence comprehension benefits from additional processing around the time of an ambiguous word. Better comprehenders may have increased availability of subordinate meanings, perhaps due to higher quality lexical representations and reflected in a positive correlation between vocabulary size and comprehension success.


The global spread of asbestos.
Asbestos continues to be used in large quantities around the world and to be an important commodity in global trade. To assess and quantify current global patterns of asbestos production, export and use; to examine global patterns of asbestos-related disease; and to examine barriers to an asbestos ban. Review of the biomedical literature describing patterns of asbestos exposure and disease; review of documents from national governments, UN agencies, and NGOs on asbestos production and use. Despite widespread knowledge of the hazards of asbestos and bans on any use of asbestos in more than 50 countries, an estimated 2 million tons of asbestos continue to be used around the world each year. Although this amount is significantly less than peak annual consumption of nearly 5 million tons two decades ago, significant amounts of asbestos are still used in India, China, Russia, and some developing countries. This use of asbestos is responsible for disease today and will cause still more asbestos-related disease in the years ahead. Real and artificially manufactured controversies regarding asbestos such as arguments about the relative hazards of different asbestos fiber types and fiber sizes have impeded bans on asbestos. All forms of asbestos pose grave dangers to human health. All are proven human carcinogens. There is no continued justification for the use of asbestos. Its production and use should be banned worldwide.


Caffeine and caffeine metabolites in relation to hypertension in U.S. adults.
Most studies assessing the association between coffee consumption and hypertension ascertained caffeine intake in terms of number of cups per days, and yield mixed results. Although the inter-individuals variability in the caffeine metabolism is known, the relation of caffeine metabolites with hypertension remains unsettled. We examined the association of caffeine and 13 direct and indirect caffeine metabolites with hypertension in U.S. adults. Using data from 2009-2010 National Health and Nutrition Examination Survey, we included 2278 individuals aged 18 to 80 years. Urinary methyluric acids (MU) and methylxanthines (MX) products of caffeine metabolism were measured using high performance liquid chromatography-electrospray ionization-tandem quadrupole mass spectrometry. We used multivariate logistic regression to model hypertension (systolic blood pressure ≥130 mmHg or diastolic blood pressure ≥80 mmHg) as functions of urinary coffee metabolites. The odds of hypertension decreased across quartiles of 3-MU, 7-MU, 3-MX and 7-MX, with 7-MU being the more powerful metabolite. Compared with adults in the bottom quartile of 7-MU, the odds of hypertension decreased by 81% (95% CI: -90 to -22%) in those in the upper quartile. In contrast, the odds ratio for being hypertensive from the bottom to the upper quartile were 4.47 (95% CI: 1.21-16.50) for 1,3-dimethyluric acid, 4.45 (95% CI: 1.48-13.39) for 1,3-dimethylxanthine, and 5.08 (95% CI: 1.11-23.36) for 1,7-dimethylxanthine. Neither insulin resistance nor abdominal obesity were moderators in these associations. Final metabolites of caffeine (namely 3-MU, 7-MU, 3-MX and 7-MX), but not caffeine, significantly reduce the odds for hypertension in this population.


Lipid mediators of tumor necrosis factor-alpha-induced uveitis.
The authors tested the hypothesis that platelet-activating factor (PAF) and cyclooxygenase metabolites of arachidonic acid mediate the ocular inflammatory response to intravitreally injected tumor necrosis factor-alpha (TNF alpha). Rabbits were treated with the PAF receptor antagonist SRI 63-441, the cyclooxygenase inhibitors indomethacin and naproxen, or SRI 63-441 and indomethacin. At 24 hr after intravitreal injection of TNF (20,000 U), the severity of inflammation was assessed based on iridal hypermia, aqueous humor leukocyte number and aqueous humor protein, immunoreactive-prostaglandin E (I-PGE), and leukotriene B4 (LTB4) concentrations. Although all of the treatments significantly reduced the severity of anterior uveitis, SRI 63-441 plus indomethacin was the most effective, indomethacin and naproxen were intermediately effective, and SRI 63-441 was the least effective. The results of this study are consistent with an important role for cyclooxygenase metabolites of arachidonic acid in the inflammatory response to TNF alpha, particularly with respect to dilation of iridal blood vessels. Although naproxen was nearly as effective as indomethacin in reducing aqueous humor I-PGE levels, indomethacin conferred significantly more protection to the blood-aqueous barrier as shown by lower protein levels in the aqueous humor. Thus, although cyclooxygenase inhibition may partially explain the protection afforded the blood-aqueous barrier by these nonsteroidal anti-inflammatory agents, the data suggest that indomethacin may also exert anti-inflammatory effects that are independent of cyclooxygenase inhibition. Furthermore, the data are consistent with TNF alpha releasing PAF in the eye and PAF acting both directly, by increasing vascular permeability, and indirectly, by promoting release of cyclooxygenase and lipoxygenase metabolites of arachidonic acid.


Study protocol for a telephone-based smoking cessation randomized controlled trial in the lung cancer screening setting: The lung screening, tobacco, and health trial.
Lung cancer mortality can be reduced by 20% via low dose CT lung cancer screening (LCS) and treatment of early-stage disease. Providing tobacco use treatment to high risk cigarette smokers in the LCS setting may result in health benefits beyond the impact of LCS. As one of the nine trials in the National Cancer Institute's Smoking Cessation at Lung Examination (SCALE) collaboration, the goal of the Lung Screening, Tobacco, and Health (LSTH) trial is to develop a scalable and cost-effective cessation intervention for subsequent implementation by LCS programs. Guided by the RE-AIM Framework, the LSTH trial is a two-arm RCT (N = 1330) enrolling English- and Spanish-speaking smokers registered for LCS at one of seven collaborating sites. Participants are randomly assigned to Usual Care (UC; three proactive telephone counseling sessions/two weeks of nicotine patches) vs. Intensive Telephone Counseling (ITC; eight proactive sessions/eight weeks of nicotine patches, plus discussion of the LCS results to increase motivation to quit). Telephone counseling is provided by tobacco treatment specialists. To increase continuity of care, referring physicians are notified of participant enrollment and smoking status following the intervention. Outcomes include: 1) self-reported 7-day, 30-day, and sustained abstinence, and biochemically-verified at 3-, 6-, and 12-months post-randomization, 2) reach and engagement of the interventions, and 3) cost-effectiveness of the interventions. The Cancer Intervention and Surveillance Modeling Network (CISNET) will model long-term impacts of six SCALE trials on the cost per life year saved, quality-adjusted life years saved, lung cancer mortality reduction, and population mortality. CLINICAL TRIALS REGISTRATION: The trial is registered at clinical trials.gov: NCT03200236.


Computational and biochemical analysis of type IV pilus dynamics and stability.
Type IV pili (T4P) are distinctive dynamic filaments at the surface of many bacteria that can rapidly extend and retract and withstand strong forces. T4P are important virulence factors in many human pathogens, including Enterohemorrhagic Escherichia coli (EHEC). The structure of the EHEC T4P has been determined by integrating nuclear magnetic resonance (NMR) and cryo-electron microscopy data. To better understand pilus assembly, stability, and function, we performed a total of 108 ms all-atom molecular dynamics simulations of wild-type and mutant T4P. Extensive characterization of the conformational landscape of T4P in different conditions of temperature, pH, and ionic strength is complemented with targeted mutagenesis and biochemical analyses. Our simulations and NMR experiments reveal a conserved set of residues defining a calcium-binding site at the interface between three pilin subunits. Calcium binding enhances T4P stability ex vivo and in vitro, supporting the role of this binding site as a potential pocket for drug design.


A Double-Blind, Randomized Clinical Trial of Niacinamide 4% versus Hydroquinone 4% in the Treatment of Melasma.
Background. Multiple modalities have been used in the treatment of melasma with variable success. Niacinamide has anti-inflammatory properties and is able to decrease the transfer of melanosomes. Objective. To evaluate the therapeutic effect of topical niacinamide versus hydroquinone (HQ) in melasma patients. Patients and Methods. Twenty-seven melasma patients were randomized to receive for eight weeks 4% niacinamide cream on one side of the face, and 4% HQ cream on the other. Sunscreen was applied along the observation period. They were assessed by noninvasive techniques for the evaluation of skin color, as well as subjective scales and histological sections initially and after the treatment with niacinamide. Results. All patients showed pigment improvement with both treatments. Colorimetric measures did not show statistical differences between both sides. However, good to excellent improvement was observed with niacinamide in 44% of patients, compared to 55% with HQ. Niacinamide reduced importantly the mast cell infiltrate and showed improvement of solar elastosis in melasma skin. Side effects were present in 18% with niacinamide versus 29% with HQ. Conclusion. Niacinamide induces a decrease in pigmentation, inflammatory infiltrate, and solar elastosis. Niacinamide is a safe and effective therapeutic agent for this condition.


Heparin induced thrombocytopenia: position paper from the Italian Society on Thrombosis and Haemostasis (SISET).
Heparin induced thrombocytopenia (HIT) is a rare immune mediated adverse drug reaction occurring after exposure to heparin. It is a serious and potentially fatal condition, which may be associated with the development of arterial or venous thrombotic events. Although known for many years, HIT is still often misdiagnosed. Pre- test clinical probability, screening for anti-PF4/heparin antibodies and documentation of their platelet activating capacity are the cornerstones of diagnosis. However, both clinical algorithms and test modalities have limited predictive values and limited diffusion so that the diagnosis and management is challenging in the clinical practice. For this reason, there is an unmet need for novel rational non-anticoagulant therapies based on the pathogenesis of HIT.The present paper reports the position of the Italian Society on Haemostasis and Thrombosis (SISET) in order to increase awareness of HIT among clinicians and other health care professionals and to provide information on the most appropriate management.


Management of obturator nevre injury during pelvic lymph node dissection.
Obturator nerve injuries may be seen during pelvic lymph node dissection in oncological surgery and although not common it is an important complication. According to the shape and location of the injury, tingling and loss of sensation may develop on the inner surface of the leg, together with loss of motor function of the adductor muscles. In this study an evaluation was made of these complications encountered in our clinic and the management strategies applied to these patients. The data were retrospectively reviewed of 843 patients who underwent open radical retropubic prostatectomy between January 2002 and May 2016. To confirm obturator nerve palsy, electrophysiological investigation (ENG-EMG) was performed immediately postoperatively and 3 weeks later. A total of 6 obturator nerve injuries occurred during pelvic lymphadenectomy (0.7%). Reapproximation end to end with sutures was applied in 3 case and sural nerve graft in 1. In the other 2 patients, just clips were placed and these were removed early during the operation. After the treatment period, neurotropic medications or physiotherapy were given in some cases according to the neurological examinations. Obturator nerve injury can be prevented by having a comprehensive knowledge of pelvic anatomy, and avoiding the use of electrocautery during lymph node dissection. The repair should be performed as soon as possible, with a tension-free reapproximation of the ends, using electrophysiological tests with a multidisciplinary approach and benefit should be taken from physiotherapy and medical treatment when needed.


Pituitary adenomas: presentations and outcomes in a South Asian country.
The purpose of this retrospective review of all operated cases of pituitary adenomas in the last decade, is to define the demographic patterns and characteristics of such tumors and to assess surgical outcomes with regards to safety and efficacy of trans-sphenoidal tumor removal in our institution. Surgically treated pituitary adenomas presenting from 1995 till 2005 were reviewed for different variables. Results were expressed as mean, standard deviation and median for continuous and number with percentage for categorical data. Chi square test was applied to measure differences and significance was taken atp value < 0.05. One hundred and twenty-five patients were operated for pituitary adenoma. Sixty-three percent were male and mean age was 37 years. Sixty percent of the patients presented with headache and/or visual symptoms. Twelve percent presented with pituitary apoplexy and 28% presented with symptoms due to pituitary hyperfunction. Fifty-five percent of patients had functioning and 44% had nonfunctioning adenomas. Mean pre operative tumor diameter from 86 pre op MRI scans was 26.76 mm (3-78 mm). Eighty-four percent of patients underwent trans-sphenoidal tumor resection and three percent had craniotomy. Mean size of post op residual tumor as calculated from 76 available post operative scans was 5.3 mm (range 0-31 mm). 17.6% of the patients required hormone replacement beyond three months and 10% were re-operated. Overall mortality was 1.6%. In Pakistan, patients are more likely to present either with apoplexy or with a giant pituitary adenoma than patients reported from developed countries. Overall, our results have been satisfactory and comparable with the literature.


Component analysis of nutritionally rich chloroplasts: recovery from conventional and unconventional green plant species.
A study of the literature indicates that chloroplasts synthesise a range of molecules, many of which have nutritional value for humans, but the nutritional credentials of chloroplasts recovered from plant cells are not established. Chloroplast-rich-fractions (CRFs) were prepared from green plant species and the macro- and micro-nutrient composition compared with the whole leaf materials (WLMs). The results indicated that, on a dry weight basis, CRF material from a range of green biomass was enriched in lipids and proteins, and in a range of micronutrients compared with the WLM. Vitamins E, pro-vitamin A, and lutein were all greater in CRF preparations. Of the minerals, iron was most notably concentrated in CRF. Spinach CRFs possessed the highest α-tocopherol [62 mg 100 g-1, dry weight (DW)], β-carotene (336 mg 100 g-1 DW) and lutein (341 mg 100 g-1 DW) contents, whilst grass CRFs had the highest concentration of alpha-linolenic acid (ALA) (69.5 mg g-1). The higher concentrations of α-tocopherol, β-carotene, lutein, ALA and trace minerals (Fe and Mn) in CRFs suggested their potential use as concentrated ingredients in food formulations deficient in these nutrients.


Functional insights into pGI2, a cryptic rolling-circle replicating plasmid from Bacillus thuringiensis.
Detailed functional analysis revealed the modular organization of pGI2, a 9672 bp plasmid from Bacillus thuringiensis H1.1 that harbours the 4149 bp transposon Tn4430. Whereas the pGI2 leading-strand replicon was identified through deletion experiments, sequence comparisons indicated the presence of an ssot-like single-strand origin commonly found among Bacillus plasmids. Southern hybridization confirmed the existence of ssDNA intermediates, but only in the case of plasmid derivatives lacking the ssot site. Moreover, the pGI2 replication protein Rep displayed significant similarity with that of pTX14-3, a 7-6 kb plasmid from B. thuringiensis serovar israelensis, suggesting that both elements are representatives of a new family of rolling-circle replicating (RCR) plasmids. In addition, both plasmids share a conserved 320 bp region downstream of their rep genes which, in the case of pGI2, appeared indispensable for replication. This region is therefore likely to correspond to, or to be part of, the actual double-strand origin of both plasmids. Another interesting feature of pGI2 is the presence of a mobilization (Mob) protein, as demonstrated by its ability to be mobilized by the conjugative plasmid pAW63 from B. thuringiensis serovar kurstaki HD73. The same transfer system was also used to unambiguously demonstrate similar properties of the related Mob-like protein from pTX14-3. A closer analysis of this family of related Mob proteins suggested a subdomanial organization among its members. Finally, the 270 residue pGI2 ORF2 was shown to be related to ORF43 of pMRC01, a 60 kb conjugative plasmid from Lactococcus lactis subsp. lactis. Although no function has been assigned to the putative ORF43 protein, it is located downstream of a bacteriocin operon, next to an IS946 element. pGI2 appears thus far as an assemblage of functional modules with no obvious metabolic function, presumably acting as a reservoir of carrier (rep and sso), rearrangement (Tn4430) or recruiting (Mob) entities for its bacterial host.


The organic solute transporters alpha and beta are induced by hypoxia in human hepatocytes.
The organic solute transporters alpha and beta (OSTα-OSTβ) form a heterodimeric transporter located at the basolateral membrane of intestinal epithelial cells and hepatocytes. Liver injury caused by ischaemia-reperfusion, cancer, inflammation or cholestasis can induce a state of hypoxia in hepatocytes. Here, we studied the effect of hypoxia on the expression of OSTα-OSTβ. OSTα-OSTβ expression was measured in Huh7 cells and primary human hepatocytes (PHH) exposed to chenodeoxycholic acid (CDCA), hypoxia or both. OSTα-OSTβ promoter activity was analysed in luciferase reporter gene assays. Binding of hypoxia-inducible factor-1 alpha (HIF-1α) to the OSTα-OSTβ gene promoters was studied in electrophoretic mobility shift assays (EMSA). Expression of OSTα and OSTβ increased in PHH under conditions of hypoxia. Exposure of Huh7 cells or PHH to CDCA (50 μM) enhanced the effect of hypoxia on OSTα mRNA levels. In luciferase assays and EMSA, the inducing effect of low oxygen could be assigned to HIF-1α, which binds to hypoxia responsive elements (HRE) in the OSTα and OSTβ gene promoters. Site-directed mutagenesis of either the predicted HRE or the bile acid responsive FXR binding site abolished inducibility of the OSTα promoter, indicating that both elements need to be intact for induction by hypoxia and CDCA. In a rat model of chronic renal failure, the known increase in hepatic OSTα expression was associated with an increase in HIF-1α protein levels. OSTα-OSTβ expression is induced by hypoxia. FXR and HIF-1α bind in close proximity to the OSTα gene promoter and produce synergistic effects on OSTα expression.


Pharmacokinetics of fenbendazole in dogs.
Fenbendazole was administered to dogs at a dose rate of 20 mg/kg body weight on a single occasion in gelatin capsules, on 5 consecutive days in feed, and on a single occasion as an alginate suspension. It was also administered at a dose rate of 100 mg/kg body weight on a single occasion in feed. Following single administration of 20 mg/kg fenbendazole mean maximum concentrations (Cmax) of the parent drug and its known active sulphoxide metabolite were 0.42 +/- 0.05 and 0.31 +/- 0.05 microgram/ml, respectively. Mean times until maximum concentrations were achieved (tmax) were 12.67 +/- 4.18 and 15.33 +/- 2.81 h, respectively, and areas under the plasma concentration-time curves (AUC) were 5.83 +/- 0.65 and 4.60 +/- 0.57 microgram.h/ml, respectively. Administration in feed increased the apparent bioavailability and administration for 5 consecutive days provided sustained plasma concentrations, generally greater than 0.2 microgram/ml. Administration as an alginate did not increase bioavailability or extend the persistence in plasma. It did increase the tmax to 16.80 +/- 2.93 and 20.00 +/- 2.53 h for fenbendazole and its sulphoxide metabolite, respectively. Increasing the dose from 20 mg/kg to 100 mg/kg did not substantially increase the Cmax or AUC.


Physician-level variation in clinical outcomes and resource use in inpatient general internal medicine: an observational study.
Variations in inpatient medical care are typically attributed to system, hospital or patient factors. Little is known about variations at the physician level within hospitals. We described the physician-level variation in clinical outcomes and resource use in general internal medicine (GIM). This was an observational study of all emergency admissions to GIM at seven hospitals in Ontario, Canada, over a 5-year period between 2010 and 2015. Physician-level variations in inpatient mortality, hospital length of stay, 30-day readmission and use of 'advanced imaging' (CT, MRI or ultrasound scans) were measured. Physicians were categorised into quartiles within each hospital for each outcome and then quartiles were pooled across all hospitals (eg, physicians in the highest quartile at each hospital were grouped together). We report absolute differences between physicians in the highest and lowest quartiles after matching admissions based on propensity scores to account for patient-level variation. The sample included 103 085 admissions to 135 attending physicians. After propensity score matching, the difference between physicians in the highest and lowest quartiles for in-hospital mortality was 2.4% (95% CI 0.6% to 4.3%, p<0.01); for readmission was 3.3% (95% CI 0.7% to 5.9%, p<0.01); for advanced imaging was 0.32 tests per admission (95% CI 0.12 to 0.52, p<0.01); and for hospital length of stay was 1.2 additional days per admission (95% CI 0.5 to 1.9, p<0.01). Physician-level differences in length of stay and imaging use were consistent across numerous sensitivity analyses and stable over time. Differences in mortality and readmission were consistent across most sensitivity analyses but were not stable over time and estimates were limited by sample size. Patient outcomes and resource use in inpatient medical care varied substantially across physicians in this study. Physician-level variations in length of stay and imaging use were unlikely to be explained by patient factors whereas differences in mortality and readmission should be interpreted with caution and could be explained by unmeasured confounders. Physician-level variations may represent practice differences that highlight quality improvement opportunities.


Fundamental principles in periodontal plastic surgery and mucosal augmentation--a narrative review.
To provide a narrative review of the current literature elaborating on fundamental principles of periodontal plastic surgical procedures. Based on a presumptive outline of the narrative review, MESH terms have been used to search the relevant literature electronically in the PubMed and Cochrane Collaboration databases. If possible, systematic reviews were included. The review is divided into three phases associated with periodontal plastic surgery: a) pre-operative phase, b) surgical procedures and c) post-surgical care. The surgical procedures were discussed in the light of a) flap design and preparation, b) flap mobilization and c) flap adaptation and stabilization. Pre-operative paradigms include the optimal plaque control and smoking counselling. Fundamental principles in surgical procedures address basic knowledge in anatomy and vascularity, leading to novel appropriate flap designs with papilla preservation. Flap mobilization based on releasing incisions can be performed up to 5 mm. Flap adaptation and stabilization depend on appropriate wound bed characteristics, undisturbed blood clot formation, revascularization and wound stability through adequate suturing. Delicate tissue handling and tension free wound closure represent prerequisites for optimal healing outcomes.


Polarized secretion of leukemia inhibitory factor.
The direction of cytokine secretion from polarized cells determines the cytokine's cellular targets. Leukemia inhibitory factor (LIF) belongs to the interleukin-6 (IL-6) family of cytokines and signals through LIFR/gp130. Three factors which may regulate the direction of LIF secretion were studied: the site of stimulation, signal peptides, and expression levels. Stimulation with IL-1beta is known to promote IL-6 secretion from the stimulated membrane (apical or basolateral) in the human intestinal epithelial cell line Caco-2. Since LIF is related to IL-6, LIF secretion was also tested in Caco-2 following IL-1beta stimulation. Signal peptides may influence the trafficking of LIF. Two isoforms of murine LIF, LIF-M and LIF-D, encode different signal peptides which have been associated with different locations of the mature protein in fibroblasts. To determine the effect of the signal peptides on LIF secretion, secretion levels were compared in Madin-Darby canine kidney (MDCK) clones which expressed murine LIF-M or LIF-D or human LIF under the control of an inducible promoter. Low and high levels of LIF expression were also compared since saturation of the apical or basolateral route would reveal specific transporters for LIF. When Caco-2 was grown on permeable supports, LIF was secreted constitutively with around 40% secreted into the apical chamber. Stimulation with IL-1beta increased LIF production. After treating the apical surface with IL-1beta, the percentage secreted apically remained similar to the untreated, whereas, when the cells were stimulated at the basolateral surface only 20% was secreted apically. In MDCK cells, an endogenous LIF-like protein was detected entirely in the apical compartment. The two mLIF isoforms showed no difference in their secretion patterns in MDCK. Interestingly, about 70% of murine and human LIF was secreted apically from MDCK over a 400-fold range of expression levels within clones and a 200,000-fold range across clones. The site of stimulation affected the polarity of LIF secretion, while, signal peptides and expression levels did not. Exogenous LIF is transported in MDCK without readily saturated steps.


The Knee Injury and Osteoarthritis Outcome Score (KOOS): a review of its psychometric properties in people undergoing total knee arthroplasty.
Systematic literature review, clinical measurement. To review and summarize the evidence regarding the psychometric properties of the Knee injury and Osteoarthritis Outcome Score (KOOS) in patients undergoing total knee arthroplasty (TKA). A comprehensive review of the existing literature was undertaken using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) systematic review guidelines. Studies were included if they reported the psychometric properties of the KOOS or the KOOS-Physical Function Shortform (KOOS-PS). Papers written in both English and German were analyzed. Studies of patients undergoing primary total TKA or TKA revision, or those with severe osteoarthritis and awaiting TKA, were considered. The methodological quality of the included articles was assessed using the Consensus-Based Standards for the Selection of Health Measurement Instruments checklist. Six studies fulfilled the inclusion criteria. Of these, 5 evaluated psychometric properties of the KOOS and 1 evaluated the KOOS-PS. Both the KOOS and KOOS-PS have moderate-to-high construct validity with existing validated self-report measures. However, construct validity of the KOOS function in sport and recreation subscale was weak. Further, these instruments demonstrated a high level of responsiveness, with effect sizes and standard response means of greater than 0.80. Overall, both questionnaires demonstrated clinically acceptable reliability (intraclass correlation coefficient of 0.70 or greater). However, somewhat lower reliability was observed for the KOOS function in sport and recreation subscale (intraclass correlation coefficients of 0.45 and 0.65, respectively) and the other symptoms subscale (internal consistency, α = .56). The KOOS and KOOS-PS exhibit clinically acceptable psychometric properties. Their strength is in large effect sizes to measure outcomes over time and their weakness is in weak-to-moderate reliability and weak construct validity in some subscales of the KOOS.


Are workplace health promotion programs effective at improving presenteeism in workers? A systematic review and best evidence synthesis of the literature.
Presenteeism is highly prevalent and costly to employers. It is defined as being present at work, but limited in some aspect of job performance by a health problem.Workplace health promotion (WHP) is a common strategy used to enhance on-the-job productivity. The primary objective is to determine if WHP programs are effective in improving presenteeism. The secondary objectives are to identify characteristics of successful programs and potential risk factors for presenteeism. The Cochrane Library, Medline, and other electronic databases were searched from 1990 to 2010. Reference lists were examined, key journals were hand-searched and experts were contacted. Included studies were original research that contained data on at least 20 participants (≥ 18 years of age), and examined the impacts of WHP programs implemented at the workplace. The Effective Public Health Practice Project Tool for Quantitative Studies was used to rate studies. 'Strong' and 'moderate' studies were abstracted into evidence tables, and a best evidence synthesis was performed. Interventions were deemed successful if they improved the outcome of interest. Their program components were identified, as were possible risk factors contributing to presenteeism. After 2,032 titles and abstracts were screened, 47 articles were reviewed, and 14 were accepted (4 strong and 10 moderate studies). These studies contained preliminary evidence for a positive effect of some WHP programs. Successful programs offered organizational leadership, health risk screening, individually tailored programs, and a supportive workplace culture. Potential risk factors contributing to presenteeism included being overweight, a poor diet, a lack of exercise, high stress, and poor relations with co-workers and management. This review is limited to English publications. A large number of reviewed studies (70%) were inadmissible due to issues of bias, thus limiting the amount of primary evidence. The uncertainties surrounding presenteeism measurement is of significant concern as a source of bias. The presenteeism literature is young and heterogeneous. There is preliminary evidence that some WHP programs can positively affect presenteeism and that certain risk factors are of importance. Future research would benefit from standard presenteeism metrics and studies conducted across a broad range of workplace settings.


Health care policy issues in the drug abuser treatment field.
As we enter the 1990s drug abuse has once again become a major health concern, and for the first time the drug treatment field has had to address many of the policy, regulation, and planning issues resulting from cost inflation that have become commonplace in other parts of the health care field. To avoid serious errors and confusion, drug abuse health policies must recognize the very different needs of the public and private sectors. The public sector, where poor addicts receive drug treatment provided or purchased by the government, has long suffered from chronically inadequate funding. Although responses to several epidemics (heroin, crack, and AIDS) have produced periods of increased allocations for drug abuse treatment, more often than not long waiting lists at programs have rationed treatment to lower-income addicts seeking care. Low salary levels have limited the quality of public treatment services, and the absence of resources has hindered the development of programs that respond to new technical developments and drug abuse problems, such as the crack epidemic. Despite severe resource shortages, the public drug treatment system has sometimes used resources inefficiently, with little attention to appropriateness of admissions, lengths of stay, ambulatory treatment modalities, or varying levels of care. Public sector goals for the 1990s should include filling current shortages in drug treatment services, developing adequate long-term funding for treating addicts who lack third-party coverage, modernizing the treatment system, developing new patterns of practice that use existing resources more efficiently, and developing a plan for treating intravenous drug users infected with the AIDS virus. In the private sector, the advent of working- and middle-class demand for drug treatment in the 1970s and 1980s has produced a new drug treatment system that suffers from many of the policy problems common to the rest of health care. Drug abuse in the workplace has resulted in much wider coverage of substance abuse services by insurance companies and HMOs. The availability of third-party funds has spawned a for-profit chemical dependency treatment industry. The high cost of private residential treatment services has caused significant cost inflation. Cost-containment measures, which are a new phenomenon for this field and are inappropriate for the public sector, have led to the same confusion and debates that they have produced in other areas of health care.(ABSTRACT TRUNCATED AT 400 WORDS)


Falls in the elderly related to postural imbalance.
Two hundred and forty-three elderly people aged 60 to 96 years were questioned about their falls, and their sway was measured. For comparison sway was also measured in 63 younger subjects. Sway increased with age and was higher in women at all ages. There was no difference in sway between those with no history of falls and those who fell only because of tripping. In both sexes sway was significantly increases in people who fell because of loss of balance and in women whose falls were due to giddiness, drop attacks, turning the head, and rising from bed or a chair. This suggests that there is a physiological decline in postural control with advancing age and also a decline due to disease of the central nervous system.


Anti-inflammatory activity of the ethanol extract of Chungkukjang, Korean fermented bean: 5-lipoxygenase inhibition.
The anti-inflammatory activity of Chungkukjang, a Korean traditional fermented soybean food (Korean-style Natto), was examined for the first time. From the results, it was found that the ethanol extract of Chungkukjang inhibited 5-lipoxygenase from A23187-treated RBL-1 cells, reducing leukoriene production (50% inhibitory concentration = 54.1 microg/mL). However, it did not greatly affect cyclooxygenase-2-catalyzed prostaglandin E(2) and inducible nitric oxide synthase-catalyzed nitric oxide production from lipopolysaccharide-treated RAW 264.7 cells at the same concentration ranges. Since leukotrienes are intimately involved in some allergic disorders, the anti-allergic activity of Chungkukjang was further examined in animal models of passive cutaneous anaphylaxis (type I hypersensitivity) and arachidonic acid-induced ear edema, two well-known in vivo models sensitive to 5-lipoxygenase inhibitors. After oral administration for 5 consecutive days, Chungkukjang significantly reduced (27.3%) passive cutaneous anaphylaxis in rats at 400 mg/kg/day. It also showed in vivo anti-inflammatory activity against arachidonic acid-induced mouse ear edema. Therefore, it is suggested that Chungkukjang may be beneficial for several allergic conditions such as asthma and atopic dermatitis.


Evaluating Models of Gesture and Speech Production for People With Aphasia.
People with aphasia use gestures not only to communicate relevant content but also to compensate for their verbal limitations. The Sketch Model (De Ruiter, 2000) assumes a flexible relationship between gesture and speech with the possibility of a compensatory use of the two modalities. In the successor of the Sketch Model, the AR-Sketch Model (De Ruiter, 2017), the relationship between iconic gestures and speech is no longer assumed to be flexible and compensatory, but instead iconic gestures are assumed to express information that is redundant to speech. In this study, we evaluated the contradictory predictions of the Sketch Model and the AR-Sketch Model using data collected from people with aphasia as well as a group of people without language impairment. We only found compensatory use of gesture in the people with aphasia, whereas the people without language impairments made very little compensatory use of gestures. Hence, the people with aphasia gestured according to the prediction of the Sketch Model, whereas the people without language impairment did not. We conclude that aphasia fundamentally changes the relationship of gesture and speech.


Cognitive Decline Over Time in Patients With Systolic Heart Failure: Insights From WARCEF.
This study sought to characterize cognitive decline (CD) over time and its predictors in patients with systolic heart failure (HF). Despite the high prevalence of CD and its impact on mortality, predictors of CD in HF have not been established. This study investigated CD in the WARCEF (Warfarin versus Aspirin in Reduced Ejection Fraction) trial, which performed yearly Mini-Mental State Examinations (MMSE) (higher scores indicate better cognitive function; e.g., normal score: 24 or higher). A longitudinal time-varying analysis was performed among pertinent covariates, including baseline MMSE and MMSE scores during follow-up, analyzed both as a continuous variable and a 2-point decrease. To account for a loss to follow-up, data at the baseline and at the 12-month visit were analyzed separately (sensitivity analysis). A total of 1,846 patients were included. In linear regression, MMSE decrease was independently associated with higher baseline MMSE score (p < 0.0001), older age (p < 0.0001), nonwhite race/ethnicity (p < 0.0001), and lower education (p < 0.0001). In logistic regression, CD was independently associated with higher baseline MMSE scores (odds ratio [OR]: 1.13; 95% confidence interval [CI]: 1.07 to 1.20]; p < 0.001), older age (OR: 1.37; 95% CI: 1.24 to 1.50; p < 0.001), nonwhite race/ethnicity (OR: 2.32; 95% CI: 1.72 to 3.13 for black; OR: 1.94; 95% CI: 1.40 to 2.69 for Hispanic vs. white; p < 0.001), lower education (p < 0.001), and New York Heart Association functional class II or higher (p = 0.03). Warfarin and other medications were not associated with CD. Similar trends were seen in the sensitivity analysis (n = 1,439). CD in HF is predicted by baseline cognitive status, demographic variables, and NYHA functional class. The possibility of intervening on some of its predictors suggests the need for the frequent assessment of cognitive function in patients with HF. (Warfarin versus Aspirin in Reduced Cardiac Ejection Fraction [WARCEF]; NCT00041938).


Inhibitory effect of green coffee bean extract on fat accumulation and body weight gain in mice.
An epidemiological study conducted in Italy indicated that coffee has the greatest antioxidant capacity among the commonly consumed beverages. Green coffee bean is rich in chlorogenic acid and its related compounds. The effect of green coffee bean extract (GCBE) on fat accumulation and body weight in mice was assessed with the objective of investigating the effect of GCBE on mild obesity. Male ddy mice were fed a standard diet containing GCBE and its principal constituents, namely, caffeine and chlorogenic acid, for 14 days. Further, hepatic triglyceride (TG) level was also investigated after consecutive administration (13 days) of GCBE and its constituents. To examine the effect of GCBE and its constituents on fat absorption, serum TG changes were evaluated in olive oil-loaded mice. In addition, to investigate the effect on hepatic TG metabolism, carnitine palmitoyltransferase (CPT) activity in mice was evaluated after consecutive ingestion (6 days) of GCBE and its constituents (caffeine, chlorogenic acid, neochlorogenic acid and feruloylquinic acid mixture). It was found that 0.5% and 1% GCBE reduced visceral fat content and body weight. Caffeine and chlorogenic acid showed a tendency to reduce visceral fat and body weight. Oral administration of GCBE (100 and 200 mg/kg. day) for 13 days showed a tendency to reduce hepatic TG in mice. In the same model, chlorogenic acid (60 mg/kg. day) reduced hepatic TG level. In mice loaded with olive oil (5 mL/kg), GCBE (200 and 400 mg/kg) and caffeine (20 and 40 mg/kg) reduced serum TG level. GCBE (1%), neochlorogenic acid (0.028% and 0.055%) and feruloylquinic acid mixture (0.081%) significantly enhanced hepatic CPT activity in mice. However, neither caffeine nor chlorogenic acid alone was found to enhance CPT activity. These results suggest that GCBE is possibly effective against weight gain and fat accumulation by inhibition of fat absorption and activation of fat metabolism in the liver. Caffeine was found to be a suppressor of fat absorption, while chlorogenic acid was found to be partially involved in the suppressive effect of GCBE that resulted in the reduction of hepatic TG level. Phenolic compounds such as neochlorogenic acid and feruloylquinic acid mixture, except chlorogenic acid, can enhance hepatic CPT activity.


GS-0976 Reduces Hepatic Steatosis and Fibrosis Markers in Patients With Nonalcoholic Fatty Liver Disease.
De novo lipogenesis is increased in livers of patients with nonalcoholic steatohepatitis (NASH). Acetyl-coenzyme carboxylase catalyzes the rate-limiting step in this process. We evaluated the safety and efficacy of GS-0976, an inhibitor of acetyl-coenzyme A carboxylase in liver, in a phase 2 randomized placebo-controlled trial of patients with NASH. We analyzed data from 126 patients with hepatic steatosis of at least 8%, based on the magnetic resonance imaging-estimated proton density fat fraction (MRI-PDFF), and liver stiffness of at least 2.5 kPa, based on magnetic resonance elastography measurement or historical biopsy result consistent with NASH and F1-F3 fibrosis. Patients were randomly assigned (2:2:1) to groups given GS-0976 20 mg, GS-0976 5 mg, or placebo daily for 12 weeks, from August 8, 2016 through July 18, 2017. Measures of hepatic steatosis, stiffness, serum markers of fibrosis, and plasma metabolomics were evaluated. The primary aims were to confirm previous findings and evaluate the relation between dose and efficacy. A relative decrease of at least 30% from baseline in MRI-PDFF (PDFF response) occurred in 48% of patients given GS-0976 20 mg (P = .004 vs placebo), 23% given GS-0976 5 mg (P = .43 vs placebo), and 15% given placebo. Median relative decreases in MRI-PDFF were greater in patients given GS-0976 20 mg (decrease of 29%) than those given placebo (decrease of 8%; P = .002). Changes in magnetic resonance elastography-measured stiffness did not differ among groups, but a dose-dependent decrease in the fibrosis marker tissue inhibitor of metalloproteinase 1 was observed in patients given GS-0976 20 mg. Plasma levels of acylcarnitine species also decreased in patients with a PDFF response given GS-0976 20 mg. GS-0976 was safe, but median relative increases of 11% and 13% in serum levels of triglycerides were observed in patients given GS-0976. In a randomized placebo-controlled trial of patients with NASH, we found 12-week administration of GS-0976 20 mg decreased hepatic steatosis, selected markers of fibrosis, and liver biochemistry. ClinicalTrials.gov ID NCT02856555.


Phase velocities of rayleigh waves in the MELT experiment on the east pacific rise
The phase velocities of Rayleigh waves increase more rapidly with distance from the East Pacific Rise (EPR) axis than is predicted by models of conductive cooling of the lithosphere. Low velocities near the axis are probably caused by partial melt at depths of 20 to 70 kilometers in a zone several hundred kilometers wide. The lowest velocities are offset to the west of the EPR. Wave propagation is anisotropic; the fast direction is approximately perpendicular to the ridge, parallel to the spreading direction. Anisotropy increases from a minimum near the axis to 3 percent or more on the flanks.


Complete resolution of advanced Mycoplasma pneumoniae encephalitis mimicking brain mass lesions: report of two pediatric cases and review of literature.
Mycoplasma pneumoniae is a well-known cause of atypical pneumonia. CNS involvement is a relatively frequent extrapulmonary manifestation, most commonly manifesting as encephalitis in the pediatric population. We present two unusual cases of M. pneumoniae encephalitis that presented with symptoms and imaging findings suggesting mass occupying lesions, and worsening altered mental status. Biopsy of the lesions was necessary in both cases to aid with diagnosis. Histopathologic features excluded neoplasm, and established the diagnosis of encephalitis, but did not point toward its etiology. The only finding that indicated M. pneumoniae as the most likely pathogen was serum IgM positivity in the absence of any other identifiable infectious source, and complete neurologic recovery following specific anti-mycoplasmal treatment. The patients were successfully treated with antibiotics and steroids, with the second case also requiring intravenous immunoglobulin and anti-epileptics. The clinical presentation and histopathologic findings suggested an immune-mediated pathogenesis, but acute disseminated encephalomyelitis was excluded due to extensive gray matter involvement. Disease resolution despite status epilepticus and herniation in case 2 is a novel finding of the study. Current principles of diagnosis and management of encephalitis as the presenting manifestation of mycoplasmal infection are discussed.


Transcriptional regulation of renal dopamine D1 receptor function during oxidative stress.
There exists a strong link between oxidative stress, renal dopaminergic system, and hypertension. It is reported that reactive oxygen species attenuate renal proximal tubular dopamine receptor (D1R) function, which disrupts sodium regulation and leads to hypertension. However, the mechanisms for renal D1R dysfunction are not clear. We investigated the role of redox-sensitive transcription factors AP1 and SP3 in transcriptional suppression of D1R gene and subsequent D1R signaling. Human kidney proximal tubular cells were treated with a pro-oxidant l-buthionine sulfoximine (BSO) with and without an antioxidant tempol. In human kidney cells, BSO caused oxidative stress and reduced D1R mRNA and membrane receptor expression. Incubation of human kidney cells with SKF38393, a D1R agonist, caused a concentration-dependent inhibition of Na/K-ATPase. However, SKF38393 failed to inhibit Na/K-ATPase in BSO-treated cells. BSO increased AP1 and SP3 nuclear expression. Transfection with AP1- or SP3-specific siRNA abolished BSO-induced D1R downregulation. Treatment of rats with BSO for 4 weeks increased oxidative stress and SP3-AP1 expression and reduced D1R numbers in renal proximal tubules. These rats exhibited high blood pressure, and SKF38393 failed to inhibit proximal tubular Na/K-ATPase activity. Control rats were kept on tap water. Tempol per se had no effect on D1R expression or other signaling molecules but prevented BSO-induced oxidative stress, SP3-AP1 upregulation, and D1R dysfunction in both human kidney cells and rats. These data show that oxidative stress via AP1-SP3 activation suppresses D1R transcription and function. Tempol mitigates oxidative stress, blocks AP1-SP3 activation, and prevents D1R dysfunction and hypertension.


Seasonal Populations of Pratylenchus penetrans and Meloidogyne hapla in Strawberry Roots.
Strawberry roots were sampled through the year to determine the populations and distribution of Pratylenchus penetrans and Meloidogyne hapla. Three strawberry root types were sampled-structural roots; feeder roots without secondary tissues; and suberized, black perennial roots. Both lesion and root-knot nematodes primarily infected feeder roots from structural roots or healthy perennial roots. Few nematodes were recovered from soil, diseased roots, or suberized roots. Lesion nematode recovery was correlated with healthy roots. In both 1997 and 1998, P. penetrans populations peaked about day 150 (end of May) and then declined. The decline in numbers corresponded to changes in total strawberry root weight and root type distribution. The loss of nematode habitat resulted from loss of roots due to disease and the transition from structural to suberized perennial roots. Meloidogyne hapla juvenile recovery peaked around 170 days (mid June) in 1997 and at 85, 147, 229, and 308 days (late March, late May, mid August, and early November, respectively) in 1998. There appear to be at least four generations per year of M. hapla in Connecticut. Diagnostic samples from an established strawberry bed may be most reliable and useful when they include feeder roots taken in late May.


[Organs, tissues, and cells donation in Mexico].
Transplants are one of the most important advances of modern medicine; in the last 50 years in our country there have been more than fifty thousand transplants, which makes it clear that this is one of the most sought-after medical practices not only in Mexico but worldwide. In life, it is possible for a person to donate a kidney, a lung or a liver segment. When brain death occurs it is possible for a person to donate kidneys, heart, liver, pancreas, intestines, lungs, blood, hematopoietic cells, bone marrow, bones, corneas, heart valves, tendons, and arteries. However, the culture of organ donation is not widespread among Mexicans, hence in our country there is not even 50 % of the number of donations recommended by WHO, which impacts the number of patients who are waiting for an organ or tissue, which causes many of them die before receiving them.


Mutation of tyrosine 383 in leukotriene A4 hydrolase allows conversion of leukotriene A4 into 5S,6S-dihydroxy-7,9-trans-11,14-cis-eicosatetraenoic acid. Implications for the epoxide hydrolase mechanism.
Leukotriene A4 hydrolase is a bifunctional zinc metalloenzyme that catalyzes the final step in the biosynthesis of the proinflammatory mediator leukotriene B4. In previous studies with site-directed mutagenesis on mouse leukotriene A4 hydrolase, we have identified Tyr-383 as a catalytic amino acid involved in the peptidase reaction. Further characterization of the mutants in position 383 revealed that [Y383H], [Y383F], and [Y383Q] leukotriene A4 hydrolases catalyzed hydrolysis of leukotriene A4 into a novel enzymatic metabolite. From analysis by high performance liquid chromatography, gas chromatography/mass spectrometry of material generated in the presence of H216O or H218O, steric analysis of the hydroxyl groups, treatment with soybean lipoxygenase, and comparison with a synthetic standard, the novel metabolite was assigned the structure 5S, 6S-dihydroxy-7,9-trans-11,14-cis-eicosatetraenoic acid (5S,6S-DHETE). The kinetic parameters for the formation of 5S,6S-DHETE and leukotriene B4 were found to be similar. Also, both activities were susceptible to suicide inactivation and were equally sensitive to inhibition by bestatin. Moreover, from the stereochemical configuration of the vicinal diol, it could be inferred that 5S, 6S-DHETE is formed via an SN1 mechanism involving a carbocation intermediate, which in turn indicates that enzymatic hydrolysis of leukotriene A4 into leukotriene B4 follows the same mechanism. Inasmuch as soluble epoxide hydrolase utilizes leukotriene A4 as substrate to produce 5S,6R-DHETE, our results also suggest a functional relationship between leukotriene A4 hydrolase and xenobiotic epoxide hydrolases.


Identification of ADAMTS7 as a novel locus for coronary atherosclerosis and association of ABO with myocardial infarction in the presence of coronary atherosclerosis: two genome-wide association studies.
We tested whether genetic factors distinctly contribute to either development of coronary atherosclerosis or, specifically, to myocardial infarction in existing coronary atherosclerosis. We did two genome-wide association studies (GWAS) with coronary angiographic phenotyping in participants of European ancestry. To identify loci that predispose to angiographic coronary artery disease (CAD), we compared individuals who had this disorder (n=12,393) with those who did not (controls, n=7383). To identify loci that predispose to myocardial infarction, we compared patients who had angiographic CAD and myocardial infarction (n=5783) with those who had angiographic CAD but no myocardial infarction (n=3644). In the comparison of patients with angiographic CAD versus controls, we identified a novel locus, ADAMTS7 (p=4·98×10(-13)). In the comparison of patients with angiographic CAD who had myocardial infarction versus those with angiographic CAD but no myocardial infarction, we identified a novel association at the ABO locus (p=7·62×10(-9)). The ABO association was attributable to the glycotransferase-deficient enzyme that encodes the ABO blood group O phenotype previously proposed to protect against myocardial infarction. Our findings indicate that specific genetic predispositions promote the development of coronary atherosclerosis whereas others lead to myocardial infarction in the presence of coronary atherosclerosis. The relation to specific CAD phenotypes might modify how novel loci are applied in personalised risk assessment and used in the development of novel therapies for CAD. The PennCath and MedStar studies were supported by the Cardiovascular Institute of the University of Pennsylvania, by the MedStar Health Research Institute at Washington Hospital Center and by a research grant from GlaxoSmithKline. The funding and support for the other cohorts contributing to the paper are described in the webappendix.


Middle East respiratory syndrome risk perception among students at a university in South Korea, 2015.
The 2015 Middle East respiratory syndrome (MERS) outbreak in South Korea was a serious threat to public health, and was exacerbated by the inappropriate responses of major institutions and the public. This study examined the sources of confusion during the MERS outbreak and identified the factors that can affect people's behavior. An online survey of the risk perception of university students in South Korea was performed after the epidemic had peaked. The questionnaire addressed the major social determinants in South Korea during the MERS epidemic. The analysis included data from 1,470 subjects who provided complete answers. The students had 53.5% of the essential knowledge about MERS. Women showed higher risk perception than men, and trust in the media was positively associated with risk perception (P < .001). Additionally, risk perception was positively associated with overreaction by the public (odds ratio, 2.80; 95% confidence interval, 2.17-3.60; P < .001). These findings suggest that media content affected the public's perception of MERS risk and that perception of a high level of risk led to overreaction. Risk perception was associated with most of the social factors examined and overreaction by the public. Therefore, providing accurate information and data to the public, establishing trust, and facilitating the development of an attitude will all be important in future crises.


Exome Sequencing of a Family with Bardet-Biedl Syndrome Identifies the Common Russian Mutation c.1967_1968delTAinsC in BBS7.
Bardet-Biedl syndrome (BBS) is a rare autosomal recessive ciliopathy characterized by obesity, postaxial polydactyly, retinitis pigmentosa, mental retardation, and kidney abnormalities. At least 19 genes have been shown to be associated with BBS, and therefore, genetic testing is highly complicated. We used an Illumina MiSeq platform for whole exome sequencing analysis of a family with strong clinical features of BBS. A homozygous c.1967_1968delTAinsC (p.Leu656fsX673; RefSeq NM_176824.2) mutation in BBS7 was identified in both affected children, while their healthy sibling and the non-consanguineous parents were heterozygous for this allele. Genotyping of 2,832 DNA samples obtained from Russian blood donors revealed 2 additional heterozygous subjects (0.07%) with the c.1967_1968delTAinsC mutation. These findings may facilitate the genetic diagnosis for Slavic BBS patients.


Problem drug use, drug misuse and deliberate self-harm: trends and patient characteristics, with a focus on young people, Oxford, 1993-2006.
Drug misuse is related to self-harm and suicide. However, relatively little is known about deliberate self-harm (DSH) in patients with drug problems and whether drug misuse by DSH patients is increasing. We used data collected by the Oxford Monitoring System for Attempted Suicide to study the characteristics of DSH patients with drug problems who presented to the general hospital in Oxford between 1993 and 2006, and who underwent psychosocial assessment at their first presentation in the study period. We also studied trends in problem drug use and drugs misused over this period. During the 14-year study period, 11,426 patients presented of whom 9,248 underwent psychosocial assessment and it was known whether or not they had a drug problem. Problem drug use was present in 805/9,248 (8.7%) patients. Problem drug use was more common in males (13.6%) than in females (5.3%). Problem drug users were younger, more likely to be socially disadvantaged, to have a personality disorder and comorbid alcohol problems and to have a further episode of DSH within a year. Problem drug use in young females was associated with higher suicidal intent scale (SIS) scores. During the study period, problem drug use and drug misuse increased in females, but not in males. Cannabis and cocaine misuse increased with time. Provision of help for DSH patients with problem drug use is particularly challenging due to their complex social and clinical characteristics and increased risk of further self-harm, suicide and accidental death. It may require extensive liaison between different services. The increasing misuse of drugs by female DSH patients and the higher SIS scores of young females are of concern.


Platelet-rich plasma in mono-segmental posterior lumbar interbody fusion.
The results of platelet-rich plasma (PRP) in spinal fusion applications are limited and controversial. Both beneficial and inhibitory effects have been shown. In this prospective randomised controlled trial, our objective was to assess both the clinical and radiological effect of PRP when added to autograft iliac crest bone in posterior lumbar interbody fusion. Forty patients were recruited for the study fulfilling strict entry requirements and were randomised with a 1:1 ratio. In each group, one patient was lost to follow-up. Thirtyeight patients completed the Visual Analogue Scale (VAS), the Oswestry Disability Index (ODI), and the Short-Form 36 (SF-36) preoperatively and postoperatively at 3, 6, 12, and 24 months, respectively. CT-scans of the lumbar spine were taken at 3, 6, and 12 months. Posterior stabilisation was achieved with pedicle screws and interbody fusion was aimed at with carbon cages filled with autologous bone. Baseline demographic data (age, sex, smoking history, preoperative outcome measures) showed no relevant difference between groups. For patients who received autograft only, the mean VAS improved by 4.0 points (p < 0.01), mean ODI improved by 32.1 points (p < 0.001), and mean SF-36 showed statistically significant improvement in each of the eight domains and in the physical (p < 0.001) and mental (p < 0.001) component summary measures. For patients who received autograft with PRP, the mean VAS improved by 4.92 points (p < 0.01), mean ODI improved by 30 points (p < 0.001), and mean SF-36 showed statistically significant improvement in six of the eight domains (p < 0.02) and in the physical (p = 0.016) and mental (p < 0.001) component summary measures. The improvement of the VAS score and the physical component summary score was more pronounced in patients who received autograft with PRP. These differences were, however, not statistically significant. CT-scans showed uneventful osseous healing in all but one patient with no difference between groups. In this prospective randomised controlled clinical and radiological trial, adding PRP in posterior lumbar interbody fusion did not lead to a substantial improvement or deterioration when compared with autologous bone only. No inhibitory effect of PRP was observed on CT-scans. From a clinical and radiological point of view, the use of PRP seems to be justified in posterior lumbar interbody fusion surgery. From an economical point of view, the expense of using PRP cannot be justified until statistical significance can be reached in a larger study.


Density dependent regulation of inflammatory responses in macrophages.
Macrophage distribution density is tightly regulated within the body, yet the importance of macrophage crowding during in vitro culture is largely unstudied. Using a human induced pluripotent stem cell (iPSC)-derived macrophage model of tissue resident macrophages, we characterize how increasing macrophage culture density changes their morphology and phenotype before and after inflammatory stimulation. In particular, density drives changes in macrophage inflammatory cytokine and chemokine secretion in both resting and activated states. This density regulated inflammatory state is also evident in blood monocyte derived-macrophages, the human monocytic THP-1 immortalized cell line, and iPSC-derived microglia. Density-dependent changes appear to be driven by a transferable soluble factor, yet the precise mechanism remains unknown. Our findings highlight cell plating density as an important but frequently overlooked consideration of in vitro macrophage research relevant to a variety of fields ranging from basic macrophage cell biology to disease studies.


Isometamidium chloride and homidium chloride fail to cure mice infected with Ethiopian Trypanosoma evansi type A and B.
Trypanosoma evansi is mechanically transmitted by biting flies and affects camels, equines, and other domestic and wild animals in which it causes a disease called surra. At least two types of Trypanosoma evansi circulate in Ethiopia: type A, which is present in Africa, Latin America and Asia, and type B, which is prevalent in Eastern Africa. Currently, no information is available about the drug sensitivity of any Ethiopian T. evansi type. This study was conducted with the objective of determining the in vivo drug sensitivity of two T. evansi type A and two type B stocks that were isolated from camels from the Tigray and Afar regions of Northern Ethiopia. We investigated the efficacy of four trypanocidal drugs to cure T. evansi infected mice: melarsamine hydrochloride (Cymelarsan), diminazene diaceturate (Veriben and Sequzene), isometamidium chloride (Veridium) and homidium chloride (Bovidium). Per experimental group, 6 mice were inoculated intraperitoneally with trypanosomes, treated at first peak parasitemia by daily drug injections for 4 consecutive days and followed-up for 60 days. Cymelarsan at 2 mg/kg and Veriben at 20 mg/kg cured all mice infected with any T. evansi stock, while Sequzene at 20 mg/kg caused relapses in all T. evansi stocks. In contrast, Veridium and Bovidium at 1 mg/kg failed to cure any T. evansi infection in mice. We conclude that mice infected with Ethiopian T. evansi can be cured with Cymelarsan and Veriben regardless of T. evansi type. In contrast, Veridium and Bovidium are not efficacious to cure any T. evansi type. Although innate resistance to phenanthridines was previously described for T. evansi type A, this report is the first study to show that this phenomenom also occurs in T. evansi type B infections.


An unusual presentation of a calcified thyroid gland.
A multinodular goitre with cystic degeneration with haemorrhage and subsequent calcification is a common occurrence in long-standing multinodular goitres. But extensive enlargement and calcification causing obstructive symptoms including dysphagia and tracheal shift has not been commonly reported in the literature. Our patient, an 82-year-old man presented with long-standing dysphagia and tracheal shift. His case proved challenging with the retrosternal location of the calcified gland, needing a collar neck incision and a manubriotomy to assess the thyroid. There was a large calcified shell to the gland that made mobilising and removing the gland very difficult. Histology revealed a large multinodular goitre with a large calcified cyst representing previous cyst haemorrhage which underwent calcification. Symptoms resolved postsurgical intervention.


NCBI GEO: archive for functional genomics data sets--update.
The Gene Expression Omnibus (GEO, http://www.ncbi.nlm.nih.gov/geo/) is an international public repository for high-throughput microarray and next-generation sequence functional genomic data sets submitted by the research community. The resource supports archiving of raw data, processed data and metadata which are indexed, cross-linked and searchable. All data are freely available for download in a variety of formats. GEO also provides several web-based tools and strategies to assist users to query, analyse and visualize data. This article reports current status and recent database developments, including the release of GEO2R, an R-based web application that helps users analyse GEO data.


Influence of resistance training combined with daily consumption of an egg-based or bagel-based breakfast on risk factors for chronic diseases in healthy untrained individuals.
Egg consumption is often discouraged due to cholesterol content; however, recent studies have not demonstrated a clear adverse influence of eggs on blood lipids. Furthermore, exercise training promotes improved lipids and blood pressure. The purpose of the study was to examine the effect of eating an isoenergetic (400 kcal) egg-based (including two eggs per day) versus bagel-based breakfasts, daily, combined with resistance training three times per week, prior to breakfast, on plasma lipids, glucose, insulin, insulin sensitivity and blood pressure in untrained individuals. Twenty-five healthy adult men and women (18-35 years of age) participated in the twelve week study following random assignment to study groups. Lipids and blood pressure were examined at baseline and after 6 and 12 weeks. Plasma triglycerides (TG) decreased significantly in the egg- based breakfast (EBB) group from baseline to six weeks (p = 0.011) and from six to twelve weeks (p = 0.045). A significant (p = 0.033) decrease in insulin sensitivity was observed in the bagel-based breakfast (BBB) group from zero to six weeks. No significant effects on systolic blood pressure, diastolic blood pressure, total cholesterol, high density lipoprotein- cholesterol or low density lipoprotein cholesterol were detected. Overall, daily breakfasts including two eggs for twelve weeks did not adversely affect lipids during a resistance training program and promoted improvements in plasma TG.


Fair Shares and Sharing Fairly: A Survey of Public Views on Open Science, Informed Consent and Participatory Research in Biobanking.
Biobanks are important resources which enable large-scale genomic research with human samples and data, raising significant ethical concerns about how participants' information is managed and shared. Three previous studies of the Canadian public's opinion about these topics have been conducted. Building on those results, an online survey representing the first study of public perceptions about biobanking spanning all Canadian provinces was conducted. Specifically, this study examined qualitative views about biobank objectives, governance structure, control and ownership of samples and data, benefit sharing, consent practices and data sharing norms, as well as additional questions and ethical concerns expressed by the public. Over half the respondents preferred to give a one-time general consent for the future sharing of their samples among researchers. Most expressed willingness for their data to be shared with the international scientific community rather than used by one or more Canadian institutions. Whereas more respondents indicated a preference for one-time general consent than any other model of consent, they constituted less than half of the total responses, revealing a lack of consensus among survey respondents regarding this question. Respondents identified biobank objectives, governance structure and accountability as the most important information to provide participants. Respondents' concerns about biobanking generally centred around the control and ownership of biological samples and data, especially with respect to potential misuse by insurers, the government and other third parties. Although almost half the respondents suggested that these should be managed by the researchers' institutions, results indicate that the public is interested in being well-informed about these projects and suggest the importance of increased involvement from participants. In conclusion, the study discusses the viability of several proposed models for informed consent, including e-governance, independent trustees and the use of exclusion clauses, in the context of these new findings about the views of the Canadian public.


Women with knee osteoarthritis have more pain and poorer function than men, but similar physical activity prior to total knee replacement.
Osteoarthritis of the knee is a major clinical problem affecting a greater proportion of women than men. Women generally report higher pain intensity at rest and greater perceived functional deficits than men. Women also perform worse than men on function measures such as the 6-minute walk and timed up and go tests. Differences in pain sensitivity, pain during function, psychosocial variables, and physical activity levels are unclear. Further the ability of various biopsychosocial variables to explain physical activity, function and pain is unknown. This study examined differences in pain, pain sensitivity, function, psychosocial variables, and physical activity between women and men with knee osteoarthritis (N = 208) immediately prior to total knee arthroplasty. We assessed: (1) pain using self-report measures and a numerical rating scale at rest and during functional tasks, (2) pain sensitivity using quantitative sensory measures, (3) function with self-report measures and specific function tasks (timed walk, maximal active flexion and extension), (4) psychosocial measures (depression, anxiety, catastrophizing, and social support), and (5) physical activity using accelerometry. The ability of these mixed variables to explain physical activity, function and pain was assessed using regression analysis. Our findings showed significant differences on pain intensity, pain sensitivity, and function tasks, but not on psychosocial measures or physical activity. Women had significantly worse pain and more impaired function than men. Their levels of depression, anxiety, pain catastrophizing, social support, and physical activity, however, did not differ significantly. Factors explaining differences in (1) pain during movement (during gait speed test) were pain at rest, knee extension, state anxiety, and pressure pain threshold; (2) function (gait speed test) were sex, age, knee extension, knee flexion opioid medications, pain duration, pain catastrophizing, body mass index (BMI), and heat pain threshold; and (3) physical activity (average metabolic equivalent tasks (METS)/day) were BMI, age, Short-Form 36 (SF-36) Physical Function, Kellgren-Lawrence osteoarthritis grade, depression, and Knee Injury and Osteoarthritis Outcome Score (KOOS) pain subscale. Women continue to be as physically active as men prior to total knee replacement even though they have significantly more pain, greater pain sensitivity, poorer perceived function, and more impairment on specific functional tasks.


Pallidal spiking activity reflects learning dynamics and predicts performance.
The basal ganglia (BG) network has been divided into interacting actor and critic components, modulating the probabilities of different state-action combinations through learning. Most models of learning and decision making in the BG focus on the roles of the striatum and its dopaminergic inputs, commonly overlooking the complexities and interactions of BG downstream nuclei. In this study, we aimed to reveal the learning-related activity of the external segment of the globus pallidus (GPe), a downstream structure whose computational role has remained relatively unexplored. Recording from monkeys engaged in a deterministic three-choice reversal learning task, we found that changes in GPe discharge rates predicted subsequent behavioral shifts on a trial-by-trial basis. Furthermore, the activity following the shift encoded whether it resulted in reward or not. The frequent changes in stimulus-outcome contingencies (i.e., reversals) allowed us to examine the learning-related neural activity and show that GPe discharge rates closely matched across-trial learning dynamics. Additionally, firing rates exhibited a linear decrease in sequences of correct responses, possibly reflecting a gradual shift from goal-directed execution to automaticity. Thus, modulations in GPe spiking activity are highest for attention-demanding aspects of behavior (i.e., switching choices) and decrease as attentional demands decline (i.e., as performance becomes automatic). These findings are contrasted with results from striatal tonically active neurons, which show none of these task-related modulations. Our results demonstrate that GPe, commonly studied in motor contexts, takes part in cognitive functions, in which movement plays a marginal role.


Metagenomic Analysis of Bacteria, Fungi, Bacteriophages, and Helminths in the Gut of Giant Pandas.
To obtain full details of gut microbiota, including bacteria, fungi, bacteriophages, and helminths, in giant pandas (GPs), we created a comprehensive microbial genome database and used metagenomic sequences to align against the database. We delineated a detailed and different gut microbiota structures of GPs. A total of 680 species of bacteria, 198 fungi, 185 bacteriophages, and 45 helminths were found. Compared with 16S rRNA sequencing, the dominant bacterium phyla not only included Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria but also Cyanobacteria and other eight phyla. Aside from Ascomycota, Basidiomycota, and Glomeromycota, Mucoromycota, and Microsporidia were the dominant fungi phyla. The bacteriophages were predominantly dsDNA Myoviridae, Siphoviridae, Podoviridae, ssDNA Inoviridae, and Microviridae. For helminths, phylum Nematoda was the dominant. In addition to previously described parasites, another 44 species of helminths were found in GPs. Also, differences in abundance of microbiota were found between the captive, semiwild, and wild GPs. A total of 1,739 genes encoding cellulase, β-glucosidase, and cellulose β-1,4-cellobiosidase were responsible for the metabolism of cellulose, and 128,707 putative glycoside hydrolase genes were found in bacteria/fungi. Taken together, the results indicated not only bacteria but also fungi, bacteriophages, and helminths were diverse in gut of giant pandas, which provided basis for the further identification of role of gut microbiota. Besides, metagenomics revealed that the bacteria/fungi in gut of GPs harbor the ability of cellulose and hemicellulose degradation.


Towards an international electronic repository and virtual laboratory of open data and open-source software for telehealth research: comparison of international, Australian and Finnish privacy policies.
Health data includes all content related to health in all data formats, document types, information systems, publication media and languages from all specialties, organisations, regions, states and countries. Capabilities to share, integrate and compare these data contents, clinical trial results and other evaluation outcomes together with telehealth applications for data processing are critical to accelerate discovery and its diffusion to clinical practice. However, the same ethical and legal frameworks that protect privacy hinder this open data and open-source code approach and the issues accumulate if moving data across national, regional or organisational borders. This can be seen as one of the reasons why many telehealth applications and health-research findings tend to be limited to very narrow domains and global results are lacking. The aim of this paper is to take steps towards establishing an international electronic repository and virtual laboratory of open data and open-source code for research purposes by comparing international, Australian and Finnish frameworks. The frameworks seem to be fundamentally similar; they apply the principles of accountability and adequacy to using and disclosing personal data. Their requirements to inform data subjects about the purposes of data collection and use before the dataset is collected, assure that individuals are no longer identifiable and to destruct data when the research activities are finished make sharing data and even secondary data difficult. Using the Internet or cloud services for sharing without proper approvals by ethics committees is technically not allowed if the data are stored in another country. The research community needs to overcome these barriers and develop a virtual laboratory, which operates on distributed data repositories. This empowers the community by enabling systematic evaluations of new technologies and research hypotheses on a rich variety of data and against existing applications, and subsequent tracking of quality improvements in time.


Local binocular fusion is involved in global binocular rivalry.
We examined whether interocular inhibition in binocular rivalry could occur at the interocular intersection of horizontal and vertical rectangular patches which are locally fusible but globally rivalrous between the two eyes. We measured contrast increment (and decrement) thresholds of a monocularly presented probe which was presented on the horizontal patch corresponding to the intersection. We found that the threshold was higher when the horizontal patch was perceptually suppressed than when it was dominant. In addition, threshold elevation did not occur when both patches were dominant, or when the horizontal patch was viewed in isolation. These results indicate that interocular inhibition occurs at the potentially fusible region, and the determination of binocular fusion or binocular rivalry does not depend on physical stimulus but rather perceptual state at the time.


Setting up a clinical trial in care homes: challenges encountered and recommendations for future research practice.
Older adults in care homes have increasingly complex health care needs, and care provision should be evidence-based whenever possible. However, recruitment of frail, older people to research is a complex process and often results in care home residents being excluded from research participation. This paper draws on the experience of setting up a randomised controlled trial to determine the effectiveness of probiotics on antibiotic-associated diarrhoea in care home residents [Probiotics for Antibiotic Associated Diarrhoea in Care Homes (PAAD) Study] in Wales. Significant challenges were encountered setting up a clinical trial in care homes. There were a number of barriers and facilitative factors encountered that were unique to this research setting. The classification of the study intervention (a widely available food supplement with a low risk safety profile) as an investigational medicinal product, with the associated requirements including obtaining statutory approvals and research governance, had a major impact. The process for setting up a clinical trial of an investigational medicinal product in care homes has been more complex and time consuming than the process for setting up an observational study in the same setting, and clinical trials in other health care settings. We recommend regulatory changes to ensure approvals processes are more proportionate to risk and context, to ensure that care home residents have the opportunity to participate in research and are able to help generate much needed evidence to underpin care. Recommendations made may inform future research practice. ISRCTN 25324586.


Differential effects of dendritic cell transfer on airway hyperresponsiveness and inflammation.
Dendritic cells (DCs) are considered to be the most efficient antigen-presenting cells. Intratracheal administration of allergen-pulsed bone marrow-derived dendritic cells (BMDCs) before allergen challenge induces airway hyperresponsiveness (AHR) and inflammation. Ovalbumin (OVA)-pulsed BMDCs from wild-type (WT) mice were transferred into naive WT, CD4(-/-), CD8(-/-), or IL-13(-/-) mice. Two days (short protocol) or 10 days (long protocol) after BMDC transfer, mice were challenged with 1% OVA for 3 days and assayed 2 days later. Transfer of OVA-primed BMDCs into BALB/c or C57BL/6 mice elicited AHR in both protocols. Airway eosinophilia, Th2 cytokines, or goblet cell metaplasia were increased in the long but not short protocol. Lung T cells from both protocols produced Th2 cytokines in response to OVA in vitro. Carboxyfluorescein diacetate succinimidylester-labeled BMDCs were observed in bronchoalveolar lavage (BAL) fluid and lung parenchyma at early time points, and were detected in draining lymph nodes 48 hours after transfer. CD8(-/-) mice developed AHR comparable to WT mice in the short protocol, but decreased levels of AHR, airway eosinophilia, Th2 cytokines in BAL fluid, and goblet cell metaplasia compared with WT mice in the long protocol. CD4(-/-) or IL-13(-/-) mice did not develop AHR or airway inflammation in either protocol. These data suggest that allergen-pulsed BMDCs initiate development of AHR that is dependent initially on CD4(+) T cells, and at later time periods on CD8+ T cells and IL-13. Thus, the timing of allergen challenge after transfer of allergen-pulsed BMDC affects the development of AHR and airway inflammation.


Combined quantum mechanical and molecular mechanical method for metal-organic frameworks: proton topologies of NU-1000.
Metal-organic frameworks (MOFs) are materials with applications in catalysis, gas separations, and storage. Quantum mechanical (QM) calculations can provide valuable guidance to understand and predict their properties. In order to make the calculations faster, rather than modeling these materials as periodic (infinite) systems, it is useful to construct finite models (called cluster models) and use subsystem methods such as fragment methods or combined quantum mechanical and molecular mechanical (QM/MM) methods. Here we employ a QM/MM methodology to study one particular MOF that has been of widespread interest because of its wide pores and good solvent and thermal stability, namely NU-1000, which contains hexanuclear zirconium nodes and 1,3,6,8-tetrakis(p-benzoic acid)pyrene (TBAPy4-) linkers. A modified version of the Bristow-Tiana-Walsh transferable force field has been developed to allow QM/MM calculations on NU-1000; we call the new parametrization the NU1T force field. We consider isomeric structures corresponding to various proton topologies of the [Zr6(μ3-O)8O8H16]8+ node of NU-1000, and we compute their relative energies using a QM/MM scheme designed for the present kind of problem. We compared the results to full quantum mechanical (QM) energy calculations and found that the QM/MM models can reproduce the full QM relative energetics (which span a range of 334 kJ mol-1) with a mean unsigned deviation (MUD) of only 2 kJ mol-1. Furthermore, we found that the structures optimized by QM/MM are nearly identical to their full QM optimized counterparts.


Dopamine affects the stability, hydration, and packing of protofibrils and fibrils of the wild type and variants of alpha-synuclein.
Parkinson's disease (PD) is characterized by the presence of cytoplasmic inclusions composed of alpha-synuclein (alpha-syn) in dopaminergic neurons. This suggests a pivotal role of dopamine (DA) on PD development. Here, we show that DA modulates differently the stability of protofibrils (PF) and fibrils (F) composed of wild type or variants of alpha-syn (A30P and A53T) as probed by high hydrostatic pressure (HHP). While in the absence of DA, all alpha-syn PF exhibited identical stability, in its presence, the variant-composed PF acquired a greater stability (DAPFwt < DAPFA30P = DAPFA53T), implying that they would last longer, which could shed light onto why these mutations are so aggressive. When alpha-syn was incubated for long times (18 days) in the presence of DA, we observed the formation of F by electronic microscopy, suggesting that the PF trapped in the presence of DA in short times can evolve into F. The stability of F was also altered by DA. DAFwt was more labile than Fwt, indicating that the former would be more susceptible to breakage. PFA30P and DAPFA30P, when added to mesencephalic and cortical neurons in culture, decreased the number and length of neurites and increased the number of apoptotic cells. Surprisingly, these toxic effects of PFA30P and DAPFA30P were practically abolished with HHP treatment, which was able to break the PF into smaller aggregates, as seen by atomic force microscopy. These results suggest that strategies aimed at breaking and/or clearing these aggregates is promising in alleviating the symptoms of PD.


Anterior chamber gas bubble emergence pattern during femtosecond LASIK-flap creation.
To characterise the emergence pattern of cavitation bubbles into the anterior chamber (AC) following femtosecond laser-assisted in situ keratomileusis (LASIK)-flap creation Retrospective review of patients undergoing femtosecond LASIK surgery at Boston Laser, a private refractive surgery practice in Boston, Massachusetts, between December 2008 and February 2014. Patient charts were reviewed to identify all cases with gas bubble migration into the AC. Surgical videos were examined and the location of bubble entry was recorded separately for right and left eyes. Five thousand one hundred and fifty-eight patients underwent femtosecond LASIK surgery. Air bubble migration into the AC, presumably via the Schlemm's canal and trabecular meshwork, occurred in 1% of cases. Patients with AC bubbles had an average age of 33±8 years with a measured LASIK flap thickness of 96±21 μm. The occurrence of gas bubbles impaired iris registration in 64% of cases. Gas bubbles appeared preferentially in the nasal or inferior quadrants for right (92% of cases) and left (100% of cases) eyes. This bubble emergence pattern is significantly different from that expected with a random distribution (p<0.0001) and did not seem associated with decentration of the femtosecond laser docking system. The migration of gas bubbles into the AC is a rare occurrence during femtosecond laser flap creation. The preferential emergence of gas bubbles into the nasal and inferior quadrants of the AC may indicate a distinctive anatomy of the nasal Schlemm's canal.