Differential brain region-specific expression of MeCP2 and BDNF in Rett Syndrome patients: a distinct grey-white matter variation.
Rett Syndrome (RTT) is a neurodevelopmental disorder caused by Methyl CpG Binding Protein 2 (MECP2) gene mutations. Previous studies of MeCP2 in the human brain showed variable and inconsistent mosaic-pattern immunolabelling, which has been interpreted as a reflection of activation-state variability. We aimed to study post mortem MeCP2 and BDNF (MeCP2 target) degradation and brain region-specific detection in relation to RTT pathophysiology. We investigated MeCP2 and BDNF stabilities in non-RTT human brains by immunohistochemical labelling and compared them in three brain regions of RTT and controls. In surgically excised samples of human hippocampus and cerebellum, MeCP2 was universally detected. There was no significantly obvious difference between males and females. However, post mortem delay in autopsy samples had substantial influence on MeCP2 detection. Immunohistochemistry studies in RTT patients showed lower MeCP2 detection in glial cells of the white matter. Glial fibrillary acidic protein (GFAP) expression was also reduced in RTT brain samples without obvious change in myelin basic protein (MBP). Neurons did not show any noticeable decrease in MeCP2 detection. BDNF immunohistochemical detection showed an astroglial/endothelial pattern without noticeable difference between RTT and controls. Our findings indicate that MeCP2 protein is widely expressed in mature human brain cells at all ages. However, our data points towards a possible white matter abnormality in RTT and highlights the importance of studying human RTT brain tissues in parallel with research on animal and cell models of RTT.


Comparison of Water Immersion Versus Air Insufflation Colonoscopy Under Various Bowel Preparation Conditions.
To investigate the differences between water immersion (WI) and air insufflation (AI) for colonoscopy under various bowel preparation conditions. In this study, 526 outpatients were randomly assigned to two groups, namely a WI group (n = 263) and an AI group (n = 263). During the procedure, the quality of bowel preparation, abdominal pain score, cecal intubation rate (CIR), adenoma detection rate (ADR), the intubation times, and other indicators were recorded. After reaching the cecum, each group of patients was subdivided into one of four grades (excellent, good, fair, and poor) according to the quality of bowel preparation. Under various bowel preparation conditions, the pain scores of the AI group were higher than those of the WI group (P < .05), but there was no significant difference between the two groups in CIR (P > .05). For the WI group compared with the AI group, the cecal intubation time (CIT) was prolonged under good bowel preparation (P = .045) and fair bowel preparation (P < .001). No significant differences were observed between the two groups on ADR in all patients (P = .476). Compared with AI colonoscopy, WI colonoscopy can decrease colonoscopy-related pain in patients for unsedated colonoscopy under various bowel preparation conditions, but there is no significant difference in CIR. WI colonoscopy requires longer CIT in patients with good and fair bowel preparation conditions. WI colonoscopy does not significantly increase ADR.


The petD gene is transcribed by functionally redundant promoters in Chlamydomonas reinhardtii chloroplasts.
FUD6, a nonphotosynthetic mutant of Chlamydomonas reinhardtii, was previously found to be deficient in the synthesis of subunit IV of the cytochrome b6/f complex, the chloroplast petD gene product (C. Lemaire, J. Girard-Bascou, F.-A. Wollman, and P. Bennoun, Biochim. Biophys. Acta 851:229-238, 1986). The lesion in FUD6 is a 236-bp deletion between two 11-bp direct repeats in the chloroplast genome. It extends from 82 to 72 bp upstream of the 5' end of wild-type petD mRNA to 156 to 166 bp downstream of the 5' end. Thus, the deletion extends into the putative promoter and 5' untranslated region of petD. No petD mRNA of the normal size can be detected in FUD6 cells, but a low level of a dicistronic message accumulates, which contains the coding regions for subunit IV and cytochrome f, the product of the upstream petA gene. petD transcriptional activity in FUD6 is not significantly altered from the wild-type level. This transcriptional activity was eliminated by petA promoter disruptions, suggesting that it originates at the petA promoter. We conclude that the petD-coding portion of most cotranscripts is rapidly degraded in FUD6, possibly following processing events that generate the 3' end of petA mRNA. A chloroplast transformant was constructed in which only the sequence from -81 to -2 relative to the major 5' end of the petD transcript was deleted. Although this deletion eliminates all detectable petD promoter activity, the transformant grows phototrophically and accumulates high levels of monocistronic petD mRNA. We conclude that the petD gene can be transcribed by functionally redundant promoters. In the absence of a functional petD promoter, a lack of transcription termination allows the downstream petD gene to be cotranscribed with the petA coding region and thereby expressed efficiently.


Clinical practice guidelines for diagnosis and treatment of chronic tinnitus in Japan.
Clinical practice guidelines in Japan for tinnitus were published in May 2019. Effective treatment of tinnitus contributes to quality of life and may improve depression, anxiety, and cognitive function. The highest priority of treatment recommended by this guideline involves educational counseling, including an explanation of the mechanisms of tinnitus. Understanding tinnitus pathology has also been reported to be a therapeutically effective educational counseling strategy. Further, explaining how sound therapy such as tinnitus retraining therapy (TRT) works is effective. Psychological and cognitive behavioral therapies may be an option. Here, the hearing aid is coupled with a sound generator in order to administer sound therapy for patients with more-severe symptoms. In Japan, it can be difficult to perform psychotherapy alongside otorhinolaryngology, and exemplary sound therapy is more likely to be carried out. In any case, the purpose of treatment is not to eliminate tinnitus, but rather reduce distress that manifests in response to tinnitus by promoting sensory adaptation. Clinically, the most important subject is not the loudness of tinnitus, but the severity of tinnitus distress. TRT consists of educational counseling and sound therapy. The tinnitus handicap inventory (THI) is used to measure the distress level for tinnitus and to determine treatment priorities. Rehabilitation of tinnitus is important because anxiety and depression are often observed in patients with severe tinnitus. Additionally, drug administration should be avoided. As a surgical treatment, a cochlear implant can affect tinnitus symptoms. It is essential to establish a treatment policy according to the disease condition and degree of distress.


Influence of renal function on the efficacy and safety of fondaparinux relative to enoxaparin in non ST-segment elevation acute coronary syndromes.
A recent randomized, controlled trial, the Fifth Organization to Assess Strategies in Acute Ischemic Syndromes (OASIS 5) trial, reported that major bleeding was 2-fold less frequent with fondaparinux than with enoxaparin in acute coronary syndromes (ACS). Renal dysfunction increases the risk for major bleeding. To compare the efficacy and safety of fondaparinux and enoxaparin over the spectrum of renal dysfunction observed in the OASIS 5 trial. Subgroup analysis of a randomized, controlled trial. Patients presenting to the hospital with non-ST-segment elevation ACS. 19,979 of the 20,078 patients in the OASIS 5 trial in whom creatinine was measured at baseline. Death, myocardial infarction, refractory ischemia, and major bleeding were evaluated separately and as a composite end point at 9, 30, and 180 days. Glomerular filtration rate (GFR) was calculated by using the Modification of Diet in Renal Disease formula. The absolute differences in favor of fondaparinux (efficacy and safety) were most marked in patients with a GFR less than 58 mL/min per 1.73 m2; the largest differences occurred in major bleeding events. At 9 days, death, myocardial infarction, or refractory ischemia occurred in 6.7% of patients receiving fondaparinux and 7.4% of those receiving enoxaparin (hazard ratio, 0.90 [95% CI, 0.73 to 1.11]); major bleeding occurred in 2.8% and 6.4%, respectively (hazard ratio, 0.42 [CI, 0.32 to 0.56]). Statistically significant differences in major bleeding persisted at 30 and 180 days. The rates of the composite end point were lower with fondaparinux than with enoxaparin in all quartiles of GFR, but the differences were statistically significant only among patients with a GFR less than 58 mL/min per 1.73 m2. Subgroup analyses warrant caution; the study was powered to detect noninferiority at 9 days. Fondaparinux is not approved for use in patients with ACS in the United States. The benefits of fondaparinux over enoxaparin when administered for non-ST-segment elevation ACS are most marked among patients with renal dysfunction and are largely explained by lower rates of major bleeding with fondaparinux.


Risk of brain infarction in familial hypercholesterolemia.
We followed 54 subjects with heterozygous familial hypercholesterolemia for an average of 10 (range 3-14) years. Half were treated surgically with partial ileal bypass and the other half (matched for age, sex, coronary heart disease, blood pressure, diabetes mellitus, smoking, obesity, and serum cholesterol concentration) were treated conservatively with diet and hypolipidemic drugs. The mean decrease in serum cholesterol concentration from the average value of 522 mg/dl on entry into the study was 32% in the surgically treated group and 10% in the conservatively treated group. One quarter of the subjects (14 of 54) had symptomatic cerebrovascular disorders and one tenth (six of 54) suffered a brain infarction at a mean age of 43 (range 30-57) years. Two thirds of the brain infarctions occurred during follow-up. The method of treatment of familial hypercholesterolemia did not affect the number of new cerebrovascular events. The incidence of brain infarction was 7.4/1000/yr. The risk of brain infarction in these subjects with familial hypercholesterolemia was at least 20 times higher than in the general population. We conclude that symptomatic subjects with familial hypercholesterolemia have not only a high risk of coronary heart disease but also a high risk of cerebrovascular disorders.


Transferring the financial risks of pharmaceutical benefits from a large health care provider in Argentina to a consortium of pharmaceutical companies.
The National Institute of Social Services for Retirees and Pensioners (NISSRP) is a nationwide health care financing agency and service provider in Argentina. Among its services, the NISSRP provides outpatient drug coverage to more than 3.3 million beneficiaries, mainly senior citizens and disabled persons. In 1997, to help cope with its rising costs, the NISSRP agreed to transfer the risk for the cost of outpatient medications and cancer-treatment drugs to a consortium of pharmaceutical companies in exchange for a fixed monthly payment. The objective of this study was to determine the impact that this new approach had on three things: (1) the level of expenditures for the medicines that were included in the agreement, (2) the pattern of nonrational prescribing for NISSRP beneficiaries, and (3) this pattern's relationship with macroeconomic variables and the pattern of prescribing for Argentina as a whole. We compared outpatient-medicine consumption in 1999 with consumption before the agreement went into effect. The actual amount that NISSRP beneficiaries spent out-of-pocket climbed from US$ 336.13 million in 1996 to US$ 473.36 million in 1999, an increase of almost 41%. The nominal amount "spent" by the NISSRP in 1999 was US$ 601.11 million, versus a real amount of US$ 374.75 million in 1996, an "increase" of 60% (that increase for the NISSRP was only theoretical since the agreement specified the fixed monthly amount that the NISSRP would have to pay to the pharmaceutical consortium). In contrast with the increased real spending by NISSRP beneficiaries, Argentina's economy remained stable over the assessed period, with the consumer price index even falling by 0.8%. We found high levels of nonrational drug use in the NISSRP system in both 1996 and 1999, indicating a serious ongoing problem. An agreement with pharmaceutical companies, like the one we have described, might add an element of financial predictability for institutions such as the NISSRP. However, such an agreement can easily result in an increased economic burden for health care beneficiaries, and without any improvement in the services that they receive. This type of agreement requires extensive mechanisms for control, follow-up, and updating, and it also risks making nonrational drug prescribing the accepted rule. While perhaps feasible, the requirements for this kind of agreement are actually very difficult to put into place, requiring additional efforts from institutions such as the NISSRP.


Disease burden of eosinophilic airway disease: Comparing severe asthma, COPD and asthma-COPD overlap.
There is less understanding of phenotypes and disease burden in asthma-COPD overlap (ACO) than either disease alone. Blood eosinophils may help identify the patients in the clinic with eosinophilic airway inflammation. The potential value of this approach requires an understanding of the illness burden associated with eosinophilic ACO, eosinophilic severe asthma and eosinophilic COPD, defined by blood eosinophils. Participants from studies of multidimensional assessment in airway disease were pooled to identify patients with ACO (n = 106), severe asthma (n = 64) and COPD alone (n = 153). Patients were assessed cross-sectionally for demographic and clinical characteristics, including disease burden indicators such as health-related quality of life (HRQoL) and past-year exacerbation. Eosinophilic patients were identified using different thresholds of blood eosinophil count. Using a blood eosinophil count ≥0.3 × 109 /L, 41% had eosinophilic airway disease: 55% in ACO, 44% in severe asthma and 29% in COPD. Blood and sputum eosinophils were moderately correlated (rs = 0.51, n = 257, P < 0.001). Burden of disease was similar between eosinophilic and non-eosinophilic airway diseases, with poor HRQoL and high number of past-year exacerbations. Burden of disease was similar across eosinophilic severe asthma, COPD and ACO. Eosinophilic COPD tended to have poorer health status than eosinophilic ACO and severe asthma; however, in context of a high prevalence of eosinophilic ACO, cumulative population-level burden of eosinophilic disease was greater in ACO. Disease burden across eosinophilic ACO, eosinophilic severe asthma and eosinophilic COPD was high, particularly cumulative population-level burden in ACO. Factors beyond airway inflammation may drive disease burden in severe patients.


Exploring the molecular basis of insecticide resistance in the dengue vector Aedes aegypti: a case study in Martinique Island (French West Indies).
The yellow fever mosquito Aedes aegypti is a major vector of dengue and hemorrhagic fevers, causing up to 100 million dengue infections every year. As there is still no medicine and efficient vaccine available, vector control largely based on insecticide treatments remains the only method to reduce dengue virus transmission. Unfortunately, vector control programs are facing operational challenges with mosquitoes becoming resistant to commonly used insecticides. Resistance of Ae. aegypti to chemical insecticides has been reported worldwide and the underlying molecular mechanisms, including the identification of enzymes involved in insecticide detoxification are not completely understood. The present paper investigates the molecular basis of insecticide resistance in a population of Ae. aegypti collected in Martinique (French West Indies). Bioassays with insecticides on adults and larvae revealed high levels of resistance to organophosphate and pyrethroid insecticides. Molecular screening for common insecticide target-site mutations showed a high frequency (71%) of the sodium channel 'knock down resistance' (kdr) mutation. Exposing mosquitoes to detoxification enzymes inhibitors prior to bioassays induced a significant increased susceptibility of mosquitoes to insecticides, revealing the presence of metabolic-based resistance mechanisms. This trend was biochemically confirmed by significant elevated activities of cytochrome P450 monooxygenases, glutathione S-transferases and carboxylesterases at both larval and adult stages. Utilization of the microarray Aedes Detox Chip containing probes for all members of detoxification and other insecticide resistance-related enzymes revealed the significant constitutive over-transcription of multiple detoxification genes at both larval and adult stages. The over-transcription of detoxification genes in the resistant strain was confirmed by using real-time quantitative RT-PCR. These results suggest that the high level of insecticide resistance found in Ae. aegypti mosquitoes from Martinique island is the consequence of both target-site and metabolic based resistance mechanisms. Insecticide resistance levels and associated mechanisms are discussed in relation with the environmental context of Martinique Island. These finding have important implications for dengue vector control in Martinique and emphasizes the need to develop new tools and strategies for maintaining an effective control of Aedes mosquito populations worldwide.


Therapy of steroid-resistant inflammatory bowel disease.
Although systemic corticosteroids are successfully administered for the induction of clinical response and remission in the majority of patients with inflammatory bowel disease (IBD) presenting with a flare, a proportion of these patients demonstrate a primary nonresponse to steroids or in the case of an initial response, they develop a resistance or a steroid dependence. Long-term therapy with corticosteroids for treatment of IBD should be avoided, given the high frequency of adverse treatment effects. Knowledge about treatment strategies in case of steroid nonresponse is therefore highly relevant. A systematic literature research was performed using Medline and Embase to summarize the currently recommended treatment strategies for steroid-resistant IBD. Treatment of steroid-resistant Crohn's disease is based on the introduction of immunomodulators such as azathioprine, 6-mercaptopurine or methotrexate, the anti-TNF drugs infliximab, adalimumab and certolizumab pegol. In the case of steroid resistance in ulcerative colitis, aminosalicylates, the above-mentioned immunomodulators, infliximab, adalimumab or calcineurin inhibitors such as ciclosporin or tacrolimus may be administered. This review summarizes the current evidence for treating steroid-resistant IBD.


Quantitative assessment of forearm muscle size, forelimb grip strength, forearm bone mineral density, and forearm bone size in determining humerus breaking strength in 10 inbred strains of mice.
Bone strength is an important clinical endpoint of osteoporosis research. The evaluation of the relative importance of bone and muscle components to bone strength has widespread implications for the understanding and preventing of osteoporosis. The objectives of this study were to understand the interrelationship between the different components of the muscular skeletal system and to determine the effect of forearm muscle size, forelimb grip strength, forearm bone mineral density (BMD), and forearm bone size on the humerus breaking strength among 10 inbred strains of mice. The forearm muscle size was measured using a peripheral quantitative computed tomography (pQCT). The forearm BMD and forearm bone size were measured using a PIXIMUS Densitometer. The forelimb grip strength and humerus breaking strength were measured using an Instron Mechanical Tester. Significant correlations were found among the five regional phenotypes. All variables have a moderately high genetic component with heritability estimates of 0.83 for forelimb grip strength, 0.76 for forearm muscle size, 0.6 for forearm BMD, 0.63 for forearm bone size, and 0.68 for humerus breaking strength. Forward stepwise multiregression analysis showed that the forearm BMD, forelimb grip strength, and forearm bone size were three major determinants of bone strength and explained 61% of the variation in bone breaking strength. These data suggest that evaluation of these three parameters together, rather than BMD alone, is a more effective, noninvasive approach for predicting fracture risk.


Laser-assisted endoscopic third ventriculostomy: long-term results in a series of 202 patients.
Endoscopic third ventriculostomy is a well-known surgical option in the treatment of noncommunicating hydrocephalus. We studied complications and long-term success in 202 patients to demonstrate the safety and efficacy of laser-assisted endoscopic third ventriculostomy (LA-ETV) using a unique "black" fiber tip/diode laser combination for controlled tissue ablation. We studied 213 LA-ETVs, which were performed in 202 patients. Patients' ages ranged from 2 days to 83 years (mean age, 27 yr). The mean follow-up period for all patients was 2.7 years (range, 2 d to 12 yr). Hydrocephalus was caused by aqueductal stenosis in 65 patients, tumors in 67 patients, hemorrhages in 24 patients, myelomeningoceles in 20 patients, cysts in 15 patients, and other causes in 11 patients. The long-term effectiveness of LA-ETV was studied with Kaplan-Meier analysis. Technically successful LA-ETVs were accomplished in 196 of the 202 patients (97%). The overall success rate for a functional LA-ETV was 68% at the 2-year follow-up evaluation. LA-ETV was more effective in patients aged 1 year and older (70% success rate) than in younger patients (59% success rate). Success rates were greater in patients with aqueductal stenosis or tumors as compared to other etiologies. Complications occurred in 22 procedures (10.3%). Only one patient (0.5%) experienced a major complication. No surgical mortalities or laser-related complications occurred. This study demonstrates that LA-ETV is a safe and effective procedure that is comparable to other techniques for ETV. LA-ETV is most effective in patients aged 1 year and older and in patients with aqueductal stenosis and tumors, with a low major complication rate.


Object-based neglect varies with egocentric position.
Different reference frames have been identified to influence neglect behavior. In particular, neglect has been demonstrated to be related to the contralesional side of the subject's body (egocentric reference frames) as well as to the contralesional side of individual objects irrespective of their position to the patient (object-based reference frame). There has been discussion whether this distinction separates neglect into body- and object-based forms. The present experiment aimed to prove possible interactions between object-based and egocentric aspects in spatial neglect. Neglect patients' eye and head movements were recorded while they explored objects at five egocentric positions along the horizontal dimension of space. The patients showed both egocentric as well as object-based behavior. Most interestingly, data analysis revealed that object-based neglect varied with egocentric position. Although the neglect of the objects' left side was strong at contralesional egocentric positions, it ameliorated at more ipsilesional egocentric positions of the objects. The patients showed steep, ramp-shaped patterns of exploration for objects located on the far contralesional side and a broadening of these patterns as the locations of the objects shifted more to the ipsilesional side. The data fitted well with the saliency curves predicted by a model of space representation, which suggests that visual input is represented in two modes simultaneously: in veridical egocentric coordinates and in within-object coordinates.


Correlates of health care satisfaction in inner-city patients with hypertension and chronic renal insufficiency.
Barriers to effective health care are potential contributors to the increased prevalence of hypertension and hypertension-related renal disease observed in black patients. We have enrolled 333 primarily elderly (mean age 69 years) black (87%) patients with hypertension and chronic renal insufficiency into a prospective randomized trial testing the effect of intense multidisciplinary management on progression of chronic renal insufficiency. These patients have an average 6 years of education and $400-$800 monthly household income: 57% have diabetes. Our baseline data include the Patient Satisfaction Questionnaire administered by home interviewers who also recorded sociodemographic data, medications and questionnaires regarding medication compliance and symptoms related to anti-hypertensive drugs. Inpatient and outpatient vital signs, test results and diagnoses came from patients' computerized medical records. We used multiple linear regression to identify correlates of overall satisfaction. We also analyzed three subscales: access to care, financial aspects and interpersonal manner of physicians. We included only variables with univariate correlations (P < 0.05) in the models. Decreased overall satisfaction correlated with more symptoms related to anti-hypertensive drugs (P < 0.001), lower medication compliance (P = 0.01), and higher diastolic blood pressure (P = 0.08). Decreased satisfaction with access to care correlated with more symptoms related to anti-hypertensive drugs (P < 0.001) and decreased medication compliance (P = 0.08). Decreased satisfaction with financial aspects of care correlated with more symptoms related to anti-hypertensive drugs (P < 0.001), lower medication compliance (P = 0.01) and more proteinuria (P = 0.02). Finally, decreased satisfaction with interpersonal manner of physicians correlated with lower medication compliance (P < 0.001), lower albumin (P = 0.01) and sodium (P = 0.04), and higher diastolic blood pressure (P = 0.04). These cross-sectional baseline data describe a group of mostly black inner-city patients with hypertension and chronic renal insufficiency in whom decreased satisfaction with care correlates with decreased medication compliance, increased symptoms related to anti-hypertensive drug therapy, higher diastolic blood pressure and more proteinuria. Our prospective study may help determine whether improving satisfaction improves compliance and blood pressure control, and forestalls complications in this high-risk population.


Moku virus; a new Iflavirus found in wasps, honey bees and Varroa.
There is an increasing global trend of emerging infectious diseases (EIDs) affecting a wide range of species, including honey bees. The global epidemic of the single stranded RNA Deformed wing virus (DWV), driven by the spread of Varroa destructor has been well documented. However, DWV is just one of many insect RNA viruses which infect a wide range of hosts. Here we report the full genome sequence of a novel Iflavirus named Moku virus (MV), discovered in the social wasp Vespula pensylvanica collected in Hawaii. The novel genome is 10,056 nucleotides long and encodes a polyprotein of 3050 amino acids. Phylogenetic analysis showed that MV is most closely related to Slow bee paralysis virus (SBPV), which is highly virulent in honey bees but rarely detected. Worryingly, MV sequences were also detected in honey bees and Varroa from the same location, suggesting that MV can also infect other hymenopteran and Acari hosts.


Content Validity of Temporal Bone Models Printed Via Inexpensive Methods and Materials.
Computed tomographic (CT) scans of the 3-D printed temporal bone models will be within 15% accuracy of the CT scans of the cadaveric temporal bones. Previous studies have evaluated the face validity of 3-D-printed temporal bone models designed to train otolaryngology residents. The purpose of the study was to determine the content validity of temporal bone models printed using inexpensive printers and materials. Four cadaveric temporal bones were randomly selected and clinical temporal bone CT scans were obtained. Models were generated using previously described methods in acrylonitrile butadiene styrene (ABS) plastic using the Makerbot Replicator 2× and Hyrel printers. Models were radiographically scanned using the same protocol as the cadaveric bones. Four images from each cadaveric CT series and four corresponding images from the model CT series were selected, and voxel values were normalized to black or white. Scan slices were compared using PixelDiff software. Gross anatomic structures were evaluated in the model scans by four board certified otolaryngologists on a 4-point scale. Mean pixel difference between the cadaver and model scans was 14.25 ± 2.30% at the four selected CT slices. Mean cortical bone width difference and mean external auditory canal width difference were 0.58 ± 0.66 mm and 0.55 ± 0.46 mm, respectively. Expert raters felt the mastoid air cells were well represented (2.5 ± 0.5), while middle ear and otic capsule structures were not accurately rendered (all averaged <1.8). These results suggest that these models would be sufficient adjuncts to cadaver temporal bones for training residents in cortical mastoidectomies, but less effective for middle ear procedures.


Changes in lymphocyte subsets in workers exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is known to have toxic effects on the haematopoietic system in animals but epidemiological studies in humans have shown inconsistent results. In this cross-sectional study we investigated changes in peripheral blood cell counts and lymphocyte subsets among workers from a Dutch historical cohort occupationally exposed to chlorophenoxy herbicides and contaminants including TCDD. Forty-seven workers who had been exposed to high levels of TCDD in the past and 38 low-exposed workers were included in the current investigation. Complete blood counts and differential and major lymphocyte subsets were analysed. Current plasma levels of TCDD (TCDD(current)) were determined by high-resolution gas chromatography/isotope-dilution high resolution mass spectrometry. TCDD blood levels at the time of last exposure (TCDD(max)) were estimated using a one-compartment first order kinetic model. Cell counts and lymphocyte subsets were similar between high- and low-exposed workers, except for a non-dose dependent increase in CD4/CD8 ratio among high-exposed workers. Interestingly, most lymphocyte subsets, in particular the B cell compartment, showed a decrease with increasing levels of both TCDD(current) and TCDD(max). Overall, our study showed that plasma TCDD levels had no effect on white blood cell counts and major subsets. However, a non-significant decrease in most lymphocyte subsets was noted, with the strongest effect for B cells. The latter finding may suggest that dioxin exposure might have an adverse impact on the haematopoietic system and lends some support to B cell lymphoma induction by dioxin.


Going Out on a Limb: Delineating The Effects of β-Branching, N-Methylation, and Side Chain Size on the Passive Permeability, Solubility, and Flexibility of Sanguinamide A Analogues.
It is well established that intramolecular hydrogen bonding and N-methylation play important roles in the passive permeability of cyclic peptides, but other structural features have been explored less intensively. Recent studies on the oral bioavailability of the cyclic heptapeptide sanguinamide A have raised the question of whether steric occlusion of polar groups via β-branching is an effective, yet untapped, tool in cyclic peptide permeability optimization. We report the structures of 17 sanguinamide A analogues designed to test the relative contributions of β-branching, N-methylation, and side chain size to passive membrane permeability and aqueous solubility. We demonstrate that β-branching has little effect on permeability compared to the effects of aliphatic carbon count and N-methylation of exposed NH groups. We highlight a new N-methylated analogue of sanguinamide A with a Leu substitution at position 2 that exhibits solvent-dependent flexibility and improved permeability over that of the natural product.


Zebrafish hoxd4a acts upstream of meis1.1 to direct vasculogenesis, angiogenesis and hematopoiesis.
Mice lacking the 4th-group paralog Hoxd4 display malformations of the anterior vertebral column, but are viable and fertile. Here, we report that zebrafish embryos having decreased function of the orthologous hoxd4a gene manifest striking perturbations in vasculogenesis, angiogenesis and primitive and definitive hematopoiesis. These defects are preceded by reduced expression of the hemangioblast markers scl1, lmo2 and fli1 within the posterior lateral plate mesoderm (PLM) at 13 hours post fertilization (hpf). Epistasis analysis revealed that hoxd4a acts upstream of meis1.1 but downstream of cdx4 as early as the shield stage in ventral-most mesoderm fated to give rise to hemangioblasts, leading us to propose that loss of hoxd4a function disrupts hemangioblast specification. These findings place hoxd4a high in a genetic hierarchy directing hemangioblast formation downstream of cdx1/cdx4 and upstream of meis1.1. An additional consequence of impaired hoxd4a and meis1.1 expression is the deregulation of multiple Hox genes implicated in vasculogenesis and hematopoiesis which may further contribute to the defects described here. Our results add to evidence implicating key roles for Hox genes in their initial phase of expression early in gastrulation.


Fast track open aortic surgery: reduced post operative stay with a goal directed pathway.
Open aortic aneurysm repair is traditionally associated with an extended hospital stay. The aim of this study was to examine the feasibility of reducing post-operative stay through the implementation of a fast-track, goal directed, clinical pathway for elective open aortic surgery. A fast-track clinical pathway for aortic surgery was introduced in a regional vascular unit in September 2005. The pathway has daily goals and targets discharge for all patients on the 3rd post operative day. This study compares thirty consecutive discharges of unselected patients undergoing elective infra-renal aortic surgery following introduction of the pathway to the thirty consecutive cases preceding its introduction. Reasons for prolonged hospital stay were recorded. Six of thirty patients achieved discharge by Day 3. The median hospital stay reduced from 9 (range 4 to 17 days) to 5 days (range 2 to 12 days) following introduction of the pathway. There was one readmission within 30 days and no complications attributable to the pathway implementation. Cardiac complications and home planning were the most common causes of delayed discharge. Post-operative stay in patients undergoing standard elective open infra-renal aortic surgery can be safely reduced with the introduction of a goal directed pathway.


An egg-adult association, gender, and reproduction in pterosaurs.
A sexually mature individual of Darwinopterus preserved together with an egg from the Jurassic of China provides direct evidence of gender in pterosaurs and insights into the reproductive biology of these extinct fliers. This new find and several other examples of Darwinopterus demonstrate that males of this pterosaur had a relatively small pelvis and a large cranial crest, whereas females had a relatively large pelvis and no crest. The ratio of egg mass to adult mass is relatively low, as in extant reptiles, and is comparable to values for squamates. A parchment-like eggshell points to burial and significant uptake of water after oviposition. This evidence for low parental investment contradicts the widespread assumption that reproduction in pterosaurs was like that of birds and shows that it was essentially like that of reptiles.


Dispatcher Stroke Recognition Using a Stroke Screening Tool: A Systematic Review.
Emergency dispatchers represent the first point of contact for patients activating an acute stroke response. Accurate dispatcher stroke recognition is associated with faster emergency medical services response time; however, stroke is often unrecognized during initial emergency calls. Stroke screening tools such as the Cincinnati Prehospital Stroke Scale have been shown to improve on-scene stroke recognition and thus have been proposed as a means to improve dispatcher accuracy. We conducted a systematic review of the accuracy of emergency dispatcher stroke recognition when employing stroke screening tools. We conducted a comprehensive search of Medline, EMBASE, CINAHL, and Cochrane databases to identify studies of dispatcher stroke recognition accuracy. Those that specifically reported dispatcher utilization of any validated stroke screening tools in isolation or in the context of a comprehensive screening algorithm such as the Medical Priority Dispatch System (MPDS) were potentially eligible. Studies that reported data sufficient for calculation of dispatcher sensitivity or positive predictive value (PPV) using a hospital-based stroke/transient ischemic attack diagnosis as the reference standard were included. Two independent reviewers determined study eligibility, assessed quality using the QUADAS 2 instrument, and abstracted data. We identified 1,413 potential studies; 54 underwent full text review. Three retrospective and 4 prospective cohort studies enrolling a total of 16,382 patients met the inclusion criteria. Stroke screening tools included MPDS (n = 4), Face Arm Speech Time (n = 2), and a novel screening algorithm developed after analysis of emergency calls for stroke (n = 1). Regardless of the screening tool employed, dispatcher stroke recognition sensitivity was suboptimal (5 studies, range 41-83%) as was the PPV (7 studies, range 42-68%). Primary study limitations included application of variable reference standards and questions regarding exclusion of subjects. No studies directly compared stroke screening algorithms and no studies specifically examined stroke recognition among potential candidates for acute stroke therapies. Even when utilizing a stroke screening tool, the accuracy of stroke recognition by emergency dispatchers was suboptimal. More research is needed to identify the causes of poor dispatcher stroke recognition and should focus on potential candidates for time-dependent stroke treatment.


Long-term effects of lifestyle intervention or metformin on diabetes development and microvascular complications over 15-year follow-up: the Diabetes Prevention Program Outcomes Study.
Effective prevention is needed to combat the worldwide epidemic of type 2 diabetes. We investigated the long-term extent of beneficial effects of lifestyle intervention and metformin on diabetes prevention, originally shown during the 3-year Diabetes Prevention Program (DPP), and assessed whether these interventions reduced diabetes-associated microvascular complications. The DPP (1996-2001) was a randomised trial comparing an intensive lifestyle intervention or masked metformin with placebo in a cohort selected to be at very high risk of developing diabetes. All participants were offered lifestyle training at the end of the DPP. 2776 (88%) of the surviving DPP cohort were followed up in the DPP Outcomes Study (DPPOS, Sept 1, 2002, to Jan 2, 2014) and analysed by intention to treat on the basis of their original DPP assignment. During DPPOS, the original lifestyle intervention group was offered lifestyle reinforcement semi-annually and the metformin group received unmasked metformin. The primary outcomes were the development of diabetes and the prevalence of microvascular disease. For the assessment of microvascular disease, we used an aggregate microvascular outcome, composed of nephropathy, retinopathy, and neuropathy. During a mean follow-up of 15 years, diabetes incidence was reduced by 27% in the lifestyle intervention group (hazard ratio 0·73, 95% CI 0·65-0·83; p<0·0001) and by 18% in the metformin group (0·82, 0·72-0·93; p=0·001), compared with the placebo group, with declining between-group differences over time. At year 15, the cumulative incidences of diabetes were 55% in the lifestyle group, 56% in the metformin group, and 62% in the placebo group. The prevalences at the end of the study of the aggregate microvascular outcome were not significantly different between the treatment groups in the total cohort (placebo 12·4%, 95% CI 11·1-13·8; metformin 13·0%, 11·7-14·5; lifestyle intervention 11·3%, 10·1-12·7). However, in women (n=1887) the lifestyle intervention was associated with a lower prevalence (8·7%, 95% CI 7·4-10·2) than in the placebo (11·0%, 9·6-12·6) and metformin (11·2%, 9·7-12·9) groups, with reductions in the lifestyle intervention group of 21% (p=0·03) compared with placebo and 22% (p=0·02) compared with metformin. Compared with participants who developed diabetes, those who did not develop diabetes had a 28% lower prevalence of microvascular complications (relative risk 0·72, 95% CI 0·63-0·83; p<0·0001). Lifestyle intervention or metformin significantly reduced diabetes development over 15 years. There were no overall differences in the aggregate microvascular outcome between treatment groups; however, those who did not develop diabetes had a lower prevalence of microvascular complications than those who did develop diabetes. This result supports the importance of diabetes prevention. National Institute of Diabetes and Digestive and Kidney Diseases.


GVI phospholipase A2 role in the stimulatory effect of sphingosine-1-phosphate on TRPC5 cationic channels.
The Transient Receptor Potential Canonical 5 (TRPC5) protein forms calcium-permeable cationic channels that are stimulated by G protein-coupled receptor agonists. The signaling pathways of such agonist effects are poorly understood. Here we investigated the potential for involvement of lysophosphatidylcholine (LPC) and arachidonic acid generated by group 6 (GVI) phospholipase A2 (PLA2) enzymes, focusing on stimulation of TRPC5 by sphingosine-1-phosphate (S1P) which acts via a pertussis toxin-sensitive (Gi/o protein) pathway without Ca2+-release. Experiments were on HEK 293 cells containing conditional expression of human TRPC5. Channel activity was recorded using an intracellular calcium indicator or whole-cell patch-clamp and PLA2 activity was detected using 3H-arachidonic acid. S1P stimulated PLA2 and TRPC5 activities. Both effects were suppressed by the GVI PLA2 inhibitor bromoenol lactone. Knock-down of GVI PLA2 by RNA interference suppressed channel activity evoked by S1P whereas activity evoked by the direct channel stimulator LPC was unaffected. Arachidonic acid did not stimulate the channels. Prior exposure of channels to LPC but not arachidonic acid suppressed channel activity evoked by S1P but not gadolinium, a putative direct stimulator of the channels. The data suggest roles of LPC and GVI PLA2 in S1P-evoked TRPC5 activity.


Spotlight on Athletes With a Disability: Malaysian Newspaper Coverage of the 2012 London Paralympic Games.
This study analyzed newspaper coverage of the 2012 London Paralympic Games by 8 Malaysian newspapers. Articles and photographs from 4 English-language and 4 Malay-language newspapers were examined from August 28 (1 day before the Games) to September 10, 2012 (1 day after the Games closing). Tables, graphs, letters, fact boxes, and lists of events were excluded from analysis. A total of 132 articles and 131 photographs were analyzed. Content analysis of the newspaper articles revealed that most (62.8%) of the articles contained positive reference to the athletes with a disability. There were equal numbers (39.1%) of action and static shots of athletes. More articles and photographs of Malaysian (58%) than non-Malaysian (42%) athletes with a disability were identified. Only 14.9% of the articles and photographs were related to female athletes with a disability.


Ruthenium-catalyzed reactions of 1-cyclopropyl-2-propyn-1-ols with anilines and water via allenylidene intermediates: selective preparation of tri- and tetrasubstituted conjugated enynes.
Ruthenium-catalyzed efficient preparation of the conjugated enynes can be carried out in the reactions of 1-cyclopropyl-2-propyn-1-ols with nitrogen- and oxygen-centered nucleophiles such as anilines and water in the presence of a catalytic amount of sulfur-bridged diruthenium complexes. The use of such complexes as catalysts realizes the completely stereoselective preparation of tri- and tetrasubstituted conjugated enynes, where ruthenium-allenylidene complexes work as key intermediates. The direct attack of nucleophiles on a cyclopropane ring connected to an allenylidene ligand is a key step to obtain the enynes stereoselectively.


Potentiation of Cytokine-Mediated Restriction of Legionella Intracellular Replication by a Dot/Icm-Translocated Effector.
Legionella pneumophila is ubiquitous in freshwater environments, where it replicates within unicellular protozoa. However, L. pneumophila is also an accidental human pathogen that can cause Legionnaires' disease in immunocompromised individuals by uncontrolled replication within alveolar macrophages. To replicate within eukaryotic phagocytes, L. pneumophila utilizes a Dot/Icm type IV secretion system to translocate a large arsenal of over 300 effector proteins directly into host cells. In mammals, translocated effectors contribute to innate immune restriction of L. pneumophila We found previously that the effector LegC4 is important for L. pneumophila replication within a natural host protist but is deleterious to replication in a mouse model of Legionnaires' disease. In the present study, we used cultured mouse primary macrophages to investigate how LegC4 attenuates L. pneumophila replication. We found that LegC4 enhanced restriction of L. pneumophila replication within macrophages activated with tumor necrosis factor (TNF) or interferon gamma (IFN-γ). In addition, expression of legC4 was sufficient to restrict Legionella longbeachae replication within TNF- or IFN-γ-activated macrophages. Thus, this study demonstrates that LegC4 contributes to L. pneumophila clearance from healthy hosts by potentiating cytokine-mediated host defense mechanisms.IMPORTANCELegionella spp. are natural pathogens of protozoa and accidental pathogens of humans. Innate immunity in healthy individuals effectively controls Legionella infection due in part to rapid and robust production of proinflammatory cytokines resulting from detection of Dot/Icm-translocated substrates, including effectors. Here, we demonstrate that the effector LegC4 enhances proinflammatory host restriction of Legionella by macrophages. These data suggest that LegC4 may augment proinflammatory signaling or antimicrobial activity of macrophages, a function that has not previously been observed for another bacterial effector. Further insight into LegC4 function will likely reveal novel mechanisms to enhance immunity against pathogens.


Evaluation of two approaches for aligning data obtained from a motion capture system and an in-shoe pressure measurement system.
An in-shoe pressure measurement (IPM) system can be used to measure center of pressure (COP) locations, and has fewer restrictions compared to the more conventional approach using a force platform. The insole of an IPM system, however, has its own coordinate system. To use an IPM system along with a motion capture system, there is thus a need to align the coordinate systems of the two measurement systems. To address this need, the current study examined two different approaches-rigid body transformation and nonlinear mapping (i.e., multilayer feed-forward neural network (MFNN))-to express COP measurements from an IPM system in the coordinate system of a motion capture system. Ten participants (five male and five female) completed several simulated manual material handling (MMH) activities, and during these activities the performance of the two approaches was assessed. Results indicated that: (1) performance varied between MMH activity types; and (2) a MFNN performed better than or comparable to the rigid body transformation, depending on the specific input variable sets used. Further, based on the results obtained, it was argued that a nonlinear mapping vs. rigid body transformation approach may be more effective to account for shoe deformation during MMH or potentially other types of physical activity.


Brain-computer interface: a reciprocal self-regulated neuromodulation.
Brain-computer interface (BCI) is a system that records brain activity and process it through a computer, allowing the individual whose activity is recorded to monitor this activity at the same time. Applications of BCIs include assistive modules for severely paralyzed patients to help them control external devices or to communicate, as well as brain biofeedback to self regulate brain activity for treating epilepsy, attention-deficit hyperactivity disorder (ADHD), anxiety, and other psychiatric conditions, or to enhance cognitive performance in healthy individuals. The vast majority of BCIs utilizes non-invasive scalp recorded electroencephalographic (EEG) signals, but other techniques like invasive intracortical EEG, or near-infrared spectroscopy measuring brain blood oxygenation are tried experimentally.


The relation of blood pressure to stroke prognosis.
The relation between blood pressure before stroke and survival after the event, was examined in the Manitoba study cohort of 3983 men. The last recorded blood pressure before the first stroke and the change in blood pressure from a measurement 5 years earlier were used. Increasing magnitude of systolic blood pressure and its 5-year changes were associated with worsening prognosis. The same association was less apparent for diastolic blood pressure and was not found for 5-year change in diastolic pressure. After adjusting for the effect of age at stroke and previous evidence of ischemic heart disease in multivariate analysis, systolic blood pressure and its 5-year change were each significant predictors of short-term (30 days) mortality. Considering all these factors as well as diastolic blood pressure, systolic blood pressure was the best predictor of short-term mortality. Thus, high blood pressure and large positive 5-year change in systolic blood pressure before stroke occurrence are significant predictors of a poor prognosis.


Primary total hip arthroplasty with a proximally porous-coated femoral stem.
The use of cementless, proximally porous-coated femoral stems for total hip arthroplasty has increased in popularity. The purpose of the present report was to examine the five to ten-year results associated with the use of a so-called second-generation circumferentially proximally porous-coated titanium-alloy stem. Between 1991 and 1994, 123 Harris-Galante Multilock femoral stems were implanted in 101 patients. The average age of the patients at the time of surgery was 53.8 years. The patients were followed prospectively and were reevaluated at a minimum of five years postoperatively. No patient was lost to follow-up. Twenty-five patients (thirty hips) were interviewed by telephone, and four patients (five hips) died during the study period because of problems that were unrelated to the operation. The remaining seventy-two patients (eighty-eight hips) had a minimum of five years of clinical and radiographic follow-up. The average duration of follow-up was seventy-eight months. At the time of the most recent follow-up, the average Harris Hip Score was 95 points. Eighty-seven (99%) of eighty-eight stems were biologically stable, with eighty-four hips (95%) having osseous ingrowth and three hips (3%) having stable fibrous fixation. One stem was revised because of loosening. Thirty-three hips (38%) had minimal proximal osteolysis, and no hip had diaphyseal osteolysis. Seventy-two hips (82%) had some degree of stress-shielding in the proximal metaphysis, but only two hips had cortical resorption. None of these patients required additional surgery, and all reported a satisfactory outcome. Given the young age and high activity level of these patients, this stem fared well: the levels of patient function and satisfaction were high, the rates of loosening and revision were very low, and distal osteolysis did not occur. Osseous fixation occurred reliably. Proximal stress-shielding was seen but did not seem to be clinically important.


Large molecules on surfaces: deposition and intramolecular STM manipulation by directional forces.
Intramolecular manipulation of single molecules on a surface with a scanning tunnelling microscope enables the controlled modification of their structure and, consequently, their physical and chemical properties. This review presents examples of intramolecular manipulation experiments with rather large molecules, driven by directional, i.e. chemical or electrostatic, forces between tip and molecule. It is shown how various regimes of forces can be explored and characterized with one and the same manipulation of a single molecule by changing the tip-surface distance. Furthermore, different deposition techniques under ultrahigh vacuum conditions are discussed because the increasing functionality of such molecules can lead to fragmentation during the heating step, making their clean deposition difficult.


Differential expression of hypoxia-inducible protein 2 among different histological types of epithelial ovarian cancer and in clear cell adenocarcinomas.
Epithelial ovarian cancer (EOC) can be classified into 5 major histological types. Among them, clear cell adenocarcinoma (CCC) has a poor response to chemotherapy and poor prognosis compared with other histological types. Previously, we reported that the hypoxia-inducible protein 2 (HIG2) gene might be a new biomarker for CCCs, based on its expression profile. In this study, we generated a polyclonal antiserum to HIG2 to explore the use of HIG2 as a predictive biomarker in EOC. In addition, HIG2 expression was evaluated in uterine endometrial and renal CCCs. Hypoxia-inducible protein 2 expression was analyzed by immunohistochemistry in formalin-fixed surgical samples from 254 EOC, 17 endometrial, and 29 renal CCC patients. Hypoxia-inducible protein 2 is expressed in 175 of 254 ovarian cancer cases. Cytoplasmic HIG2 expression is significantly more frequent in ovarian CCC (83.1%) than in serous (54.9%, P = 0.0001), mucinous (40%, P = 0.00002), or endometrioid (58.1%, P = 0.003) adenocarcinoma. The chemoresponse rate was higher in 24 ovarian CCC patients with cytoplasmic HIG2 expression than in 6 CCC patients without HIG2 expression (62.5% [15/24] vs 0% [0/6], P = 0.02). In contrast, there was no relationship between nuclear HIG2 expression and chemoresponse. Cytoplasmic and nuclear HIG2 expressions are significantly more frequent in ovarian and uterine than renal CCC (P = 0.04). Hypoxia-inducible protein 2 may be used as a marker for early detection of ovarian CCCs or for prediction of response to chemotherapy, but HIG2 expression does not predict survival of patients with CCC.


Cyst fluid interleukin-1beta (IL1beta) levels predict the risk of carcinoma in intraductal papillary mucinous neoplasms of the pancreas.
Biomarkers for high-grade dysplasia in patients with radiographically identified intraductal papillary mucinous neoplasms (IPMN) have not been described. We hypothesized that dysplasia in IPMN invokes an immunogenic/proinflammatory microenvironment that can be identified by cyst fluid cytokine levels. Pancreatic cyst fluid aspirates were collected at resection (2005-2009). Samples were grouped into low-risk [low-grade (n = 6) or moderate dysplasia (n = 15)] and high-risk groups [high-grade dysplasia (n = 13) or carcinoma (n = 6)]. Cytokine expression was determined using a multiplex sandwich immunoassay. Differences in cytokine expression were evaluated using the 2-sample t test. Sample classification was performed using a logistic regression adjusting for sample covariates. IL5 and IL8 concentrations were higher in the cyst fluid from patients in the high-risk group than the low-risk group. Interleukin (IL)-1β concentrations were also higher in the cyst fluid from patients with high-grade dysplasia or cancer (n = 19) than those with low- or moderate-grade dysplasia (n = 21, 539 ± 255 pg/mL vs. 0.2 ± 0.1 pg/mL; P < 0.0001). IL1β remained a significant predictor of high-risk cysts after multivariate analysis. There was no significant difference in levels of IL2, IL4, IL10, IL12, IL13, TNF-α, or IFN-γ between the groups. That IL1β levels identified cysts at a high risk of malignancy was confirmed in an independent validation set. Cyst fluid levels of IL1β can differentiate low- from high-risk IPMN. This study introduces IL1β as a potential biomarker for validation in larger clinical studies.


Biomimetic collagen-hydroxyapatite composite fabricated via a novel perfusion-flow mineralization technique.
Prevalent three-dimensional scaffolds for bone tissue engineering are mineralized collagen-hydroxyapatite (Col/HA) composites. Conventional mineralization techniques are either to coat collagen scaffold surfaces with minerals or to simply mix collagen and mineral nanoparticles together. These conventional in vitro collagen mineralization methods are different from the in vivo bone formation process and often result in scaffolds that are not suitable for bone tissue engineering. In this study, a unique perfusion-flow (i.e., dynamic) in conjunction with a previously described polymer-induced liquid-precursor (PILP) method was used to fabricate a porous Col/HA composite. The dynamic flow emulated the physiological extracellular fluid flow containing the mineralization ions, while the PILP method facilitated the deposition of the HA crystals within the collagen fibrils (i.e., intrafibrillar mineralization). By utilizing a dynamic PILP technique to mimic the in vivo bone formation process, the resultant Col/HA composite has a similar structure and compositions like human trabecular bone. A comparison of the dynamic and static mineralization methods revealed that the novel dynamic technique facilitates more efficient and homogenous mineral deposition throughout the Col/HA composite. The dynamic intrafibrillar mineralization method generated stiff Col/HA composites with excellent surface property for cell attachment and growth. The human mesenchymal stem cells cultured on the Col/HA composites quickly remodeled the scaffolds and resulted in constructs with an extensive cell-derived extracellular matrix network.


Endothelial Colony Forming Cells as an Autologous Model to Study Endothelial Dysfunction in Patients with a Bicuspid Aortic Valve.
Bicuspid aortic valve (BAV), the most common congenital heart defect, is associated with an increased prevalence of aortic dilation, aortic rupture and aortic valve calcification. Endothelial cells (ECs) play a major role in vessel wall integrity. Little is known regarding EC function in BAV patients due to lack of patient derived primary ECs. Endothelial colony forming cells (ECFCs) have been reported to be a valid surrogate model for several cardiovascular pathologies, thereby facilitating an in vitro system to assess patient-specific endothelial dysfunction. Therefore, the aim of this study was to investigate cellular functions in ECFCs isolated from BAV patients. Outgrowth and proliferation of ECFCs from patients with BAV (n = 34) and controls with a tricuspid aortic valve (TAV, n = 10) were determined and related to patient characteristics. Interestingly, we were only able to generate ECFCs from TAV and BAV patients without aortic dilation, and failed to isolate ECFC colonies from patients with a dilated aorta. Analyzing EC function showed that while proliferation, cell size and endothelial-to-mesenchymal transition were similar in TAV and BAV ECFCs, migration and the wound healing capacity of BAV ECFCs is significantly higher compared to TAV ECFCs. Furthermore, calcification is blunted in BAV compared to TAV ECFCs. Our results reveal ECs dysfunction in BAV patients and future research is required to unravel the underlying mechanisms and to further validate ECFCs as a patient-specific in vitro model for BAV.


Effects of levosimendan on circulating pro-inflammatory cytokines and soluble apoptosis mediators in patients with decompensated advanced heart failure.
This randomized, placebo-controlled trial showed that levosimendan administration causes a significant reduction of circulating proinflammatory cytokine interleukin-6 and soluble apoptosis mediators, such as soluble Fas and Fas ligand in patients with decompensated heart failure. These immunomodulatory effects may lead to improvement of symptoms and echocardiographic markers of cardiac contractile performance in these patients.


Thoracic Surgery Directors Association Award. What is the optimal management of late-presenting survivors of acute type A aortic dissection?
Although type A aortic dissections represent a surgical emergency, some patients present late after the onset of symptoms. Optimal management of this cohort has not been defined. Data on 195 patients with type A dissections followed up at a single institution between 1985 and 2005 were collected prospectively. Of these, 93 patients (47.2%) presented 48 hours or later after the initial onset of pain (group A), and the remaining 102 patients underwent immediate operative repair (group B). Median follow-up was 41.8 months (range, 0 to 386 months). Patients in group A were older (68.8 versus 59.3 years, p = 0.0005) and had a higher incidence of coronary artery disease (42.5% versus 14.6%, p < 0.0001), pulmonary disease (26.6% versus 8.4%, p = 0.0023), and congestive heart failure (14.1% versus 1.0%, p = 0.0004). Long-term survival was similar, although group B showed a trend toward improved 30-day mortality (16.5% versus 8.7%, p = 0.1035). Of the 92 patients in group A, 53 (57.6%) eventually underwent operative repair a median of 8.2 days after symptom onset. There was a trend toward improved long-term survival among patients undergoing repair (p = 0.1031). Initial medical management with interval operative repair of selected patients referred greater than 2 days following an acute type A dissection is a viable option. Delayed repair after optimization of the clinical condition and detailed evaluation of concomitant diseases results in excellent long-term results.


Dopaminergic nature of acute cathine tolerance.
Cathine is a psychoactive constituent in the leaves of the Khat shrub which are habitually ingested for their stimulatory effects in many parts of the world. Rats were trained to discriminate the stimulus effect of intraperitoneally administered 4.8 mg/kg d-cathine and, once trained, administration of another Khat constituent, cathinone, was shown to produce cathine-like effects. This generalization to cathinone was dose-responsive when testing occurred 24 hr after vehicle administration, whereas prior administration of cathine resulted in a diminished discriminative response to subsequent cathinone administration possibly as a result of the development of acute tolerance. CGS 10746B, a compound that blocks presynaptic release of dopamine, significantly decreased rats' ability to discriminate cathine when it was administered 25 min prior to cathine testing and it reversed the acute tolerance observed when cathine was tested 24 hr after cathine administration. These results indicate that a previously reported acute tolerance effect to cathine after cathinone administration in cathinone-trained rats appears to be symmetrical in that there is acute tolerance to cathinone after cathine in these cathine-trained rats. The results with CGS 10746B would suggest that both the cathine-induced discriminative cue and cathine's ability to produce acute tolerance are mediated by presynaptic dopamine release.


Molecular population genetics of floral homeotic loci. Departures from the equilibrium-neutral model at the APETALA3 and PISTILLATA genes of Arabidopsis thaliana.
Molecular variation in genes that regulate development provides insights into the evolutionary processes that shape the diversification of morphogenetic pathways. Intraspecific sequence variation at the APETALA3 and PISTILLATA floral homeotic genes of Arabidopsis thaliana was analyzed to infer the extent and nature of diversity at these regulatory loci. Comparison of AP3 and PI diversity with three previously studied genes revealed several features in the patterning of nucleotide polymorphisms common between Arabidopsis nuclear loci, including an excess of low-frequency nucleotide polymorphisms and significantly elevated levels of intraspecific replacement variation. This pattern suggests that A. thaliana has undergone recent, rapid population expansion and now exists in small, inbred subpopulations. The elevated intraspecific replacement levels may thus represent slightly deleterious polymorphisms that differentiate distinct ecotypes. The distribution of replacement and synonymous changes in AP3 and PI core and noncore functional domains also indicates differences in the patterns of molecular evolution between these interacting floral regulatory genes.


[Relationship between serum albumin level and prognosis in children with sepsis, severe sepsis or septic shock].
Sepsis, severe sepsis, and septic shock are frequently encountered and highly lethal conditions among children treated in ICU; however, no many reports on research on serum albumin levels of children under these conditions are available in literature and little is known about the relationship between serum albumin level and prognosis. In this prospective study, the author observed the levels of serum albumin of children with sepsis/severe sepsis/septic shock, and explored the relationship between serum albumin level and severity of illness or prognosis. For sepsis/severe sepsis/septic shock patients admitted to the pediatric intensive care unit (PICU) during the year 2008, serum albumin concentration in peripheral blood was examined within 24 h after admission. According to the diagnostic criteria, hypoalbuminemia was defined as serum albumin level of < 35 g/L. A total of 247 cases of sepsis/severe sepsis/septic shock children were enrolled. In the order of sepsis/severe sepsis/septic shock group, there were 143, 65 and 39 cases. Overall rate of hypoalbuminemia was 72.9% (180/247). The rate of hypoalbuminemia in children with sepsis, severe sepsis or septic shock was 59.4% (85/143), 86.2% (56/65) and 100% (39/39), respectively. The mortality of hypoalbuminemic patients in sepsis, severe sepsis and septic shock group showed significant difference (P < 0.001). The rates of hypoalbuminemia of the survived cases (69.5%) was significantly lower than that of the non-survived cases (94.1%). Pediatrics critical illness score (PCIS) and serum albumin concentration was positively correlated (P < 0.001), and the mortality was negatively correlated with serum albumin concentration (P < 0.05). Hypoalbuminemia is common among children with sepsis/severe sepsis/septic shock and serum albumin level is closely related to prognosis. Serum albumin level monitoring on the basis of PCIS scoring has important clinical prognostic significance in evaluating the prognosis of severe sepsis/septic shock patients.


Characterisation of white line degeneration in sheep and evidence for genetic influences on its occurrence.
Shelly hoof in sheep occurs when the hoof wall becomes detached from the laminar corium; it often then becomes impacted with debris leading to infection, pain and lameness. The problem of shelly hoof is under-reported and is often confused with classical footrot. A study was conducted using data on 9,169 Blackface and Texel sheep from 22 farms in the UK. The aim of the study was to estimate the prevalence of shelly hoof in these breeds, to quantify the genetic basis to shelly hoof, and to describe a pilot trail to characterise the physical properties of horn using Transmission Electron Microscopy (TEM). The results showed that shelly hoof has a high prevalence (47% for Blackface and 24% for Texel ewes) and is under moderate genetic control (h(2) = 0.3). The TEM images showed fragmented dorsal wall horn, microscopic crevices leading deeper into the hoof accompanied by bacteria and other micro-organisms. The invasion of bacteria was facilitated by separation of the cells in a characteristic 'un-zipping' phenomenon, poor attachment of the keratin to the cell membranes, poor cell membrane quality with a 'lace-chain' effect and incomplete keratinisation of the cells.


Effectiveness of High-Fidelity Simulation in Training Emergency Medicine Physicians in Point of Care Ultrasonography in Pakistan: A Quasi-Experimental Study.
Background Point-of-care ultrasound (PoCUS) is frequently utilized in emergency medicine (EM), with an extended-focused assessment with sonography in trauma (e-FAST) being the most widely used PoCUS modality. This modality is not only time- and cost-efficient, but it is highly accurate in the diagnosis and management of surgical patients in the emergency department, as well as being highly predictive of patient outcomes. Targeted training is essential to ensure a learner's confidence in image acquisition, interpretation, and translation of knowledge to making clinical decisions. High-fidelity simulation offers a uniquely safe and "mistake-forgiving" environment to teach and train medical professionals. The present study evaluated the effectiveness of a high-fidelity simulator to train EM physicians in e-FAST at a tertiary care teaching hospital in a lower-middle-income country. Methods This quasi-experimental study was performed at a state-of-the-art simulation center of a multidisciplinary university hospital in Karachi, Pakistan. Subjects were included if they were EM physicians who volunteered to participate and were available for the entire training and testing period. The educational intervention included lectures and hands-on practice on a high-fidelity simulator (SonoSim, Santa Monica, CA). Knowledge and image interpretation on e-FAST were evaluated using a questionnaire, administered before and after the training course. Each participant's ability to acquire and interpret satisfactory images was assessed by experienced EM physicians and recorded. Participants were also administered a needs assessment survey and a course evaluation. Data were analyzed using IBM SPSS Statistics for Windows, Version 20.0 (Armonk, NY: IBM Corp.). All the tests were two-sided, and p-values ≤0.05 were considered significant. Baseline characteristics and outcome variables were recorded and compared by Wilcoxon signed-rank tests. Results A total of 31 EM physicians, 12 (38.7%) men and 19 (61.3%) women, were enrolled in the study, with 24 (77.3%) having one to three years of EM experience. Mean and percentage group performance improved from 6 and 40% before the intervention to 14.5 and 96.6% after the intervention (Z=4.867, p≤0.05). Most improvement in image acquisition on high-fidelity simulation was observed in the upper right quadrant of the suprapubic window (29/31; 93.5%), followed by the upper left quadrant (27/31; 87%) and the subxiphoid window (21/31; 67%). All 31 participants reported improvements in comfort and confidence level with PoCUS after attending the workshop. Conclusions EM physicians who attended a brief workshop incorporating simulation demonstrated improvements in knowledge and image acquisition skills in all domains tested. High-fidelity simulation training is an effective modality for training EM physicians in e-FAST.


Increased permeability of cutaneous lymphatic capillaries and enhanced blood flow in psoriatic plaques.
Morphological abnormalities of microvessels are described in psoriasis. However, there are conflicting data as to whether their function is also altered. Our aim was to study the morphology and function of the lymphatic capillaries of psoriatic skin. Morphology and permeability of initial lymphatics were studied by microlymphography and densitometry in 20 patients. Perfusion was studied by laser Doppler fluxmetry. Permeability of lymphatics in plaques was increased by 7.6% compared to unafflicted skin (p < 0.001). Lymphatic vessel density and the extension of dye in lymphatic networks were not significantly different between involved and uninvolved areas. Both sites showed a wide range of diameters of lymphatics. The median laser Doppler flux in plaques was increased by 144% (91-380%) compared to unaffected skin (p < 0.001). Increased permeability of lymphatics and increased blood flow was demonstrated in vivo in psoriatic skin lesions. These findings may reflect the local inflammatory process and may be used as markers when studying new therapeutic approaches for psoriasis.


Localization of transcripts from the wingless gene in whole Drosophila embryos.
In situ hybridization has been used to detect transcription in whole Drosophila embryos. Improved results have been obtained by incubating the hybridized embryos in liquid emulsion prior to autoradiographic exposure. This technique has been used to map the distribution of transcripts from the segment-polarity gene wingless (wg), which is transcribed in a stripe in each segment of the trunk region. By the extended germband stage additional areas of transcription in the head and caudal regions define a total of 21 domains, comprising the foregut, six regions in the head, three thoracic and ten abdominal segments, and the hindgut. At the end of the extended germband stage, the pattern of wg transcription is no longer uniform in the dorsoventral axis: wg transcripts are absent from the lateral epidermis. This pattern of wg transcription is discussed with particular regard to the segmentation of the terminalia. Also it is suggested that the dorsoventral reorganization could be related to the specification of the imaginal disc primordia.


Role of systolic blood pressure and plasma triglycerides in diabetic peripheral arterial disease. The Edinburgh Artery Study.
To determine the risk factors for peripheral arterial disease (PAD) in a diabetic population and to examine whether different levels of these risk factors might explain why diabetic subjects have an increased risk of PAD compared with normal glucose tolerance subjects. There were 1,592 men and women aged 55-74 years selected at random from the age-sex registers of 11 general practices in Edinburgh, Scotland. Subjects underwent a comprehensive medical examination, including assessment for PAD (intermittent claudication on World Health Organization questionnaire or major asymptomatic disease on noninvasive testing) and a glucose tolerance test. Of the subjects, 288 (18.7%) were found to have diabetes or impaired glucose tolerance (IGT). The prevalence of PAD was greater in those with diabetes/IGT (20.6%) compared with those with normal glucose tolerance (12.5%) (odds ratio [OR] 1.64, 95% CI 1.17-2.31). Among the diabetes/IGT group, mean levels of smoking, systolic blood pressure, and triglycerides were higher in subjects with PAD than in those without PAD (P < or = 0.05). Mean levels of systolic blood pressure and plasma triglycerides were also higher in diabetic subjects than in nondiabetic subjects with PAD (P < or = 0.05). In multivariate analysis, those with diabetes/IGT no longer had a significantly higher risk of PAD after adjusting separately for systolic blood pressure (OR 1.22, 95% CI 0.85-1.73) and plasma triglycerides (OR 1.26, 95% CI 0.89-1.79). Simultaneous adjustment for both systolic blood pressure and triglycerides reduced the risk of PAD among diabetic subjects to 1.11 (95% CI 0.78-1.58). Increased mean levels of triglycerides and systolic blood pressure may help to explain the higher prevalence of PAD in diabetic subjects compared with that in normal glucose tolerance subjects.


Enhanced production of IL-10 by dendritic cells deficient in CIITA.
Dendritic cells (DC) are professional APCs that play a critical role in regulating immunity. In DC, maturation-induced changes in MHC class II expression and Ag presentation require transcriptional regulation by CIITA. To study the role of CIITA in DC, we evaluated key cell functions in DC from CIITA-deficient (CIITA(-/-)) mice. The ability to take up Ag, measured by fluid phase endocytosis, was comparable between CIITA(-/-) and control DC. Although CIITA(-/-) DC lack MHC class II, they maintained normal expression of costimulatory molecules CD80, CD86, and CD40. In contrast, CIITA(-/-) DC activated with LPS or CpG expressed increased IL-10 levels, but normal levels of TNF-alpha and IL-12 relative to control. Enhanced IL-10 was due to greater IL-10 mRNA in CIITA(-/-) DC. Abeta(-/-) DC, which lack MHC class II but express CIITA normally, had exhibited no difference in IL-10 compared with control. When CIITA was cotransfected with an IL-10 promoter-reporter into a mouse monocyte cell line, RAW 264.7, IL-10 promoter activity was decreased. In addition, reintroducing CIITA into CIITA(-/-) DC reduced production of IL-10. In all, these data suggest that CIITA negatively regulates expression of IL-10, and that CIITA may direct DC function in ways that extend beyond control of MHC class II.


Angiotensin II stimulates H⁺-ATPase activity in intercalated cells from isolated mouse connecting tubules and cortical collecting ducts.
Intercalated cells in the collecting duct system express V-type H(+)-ATPases which participate in acid extrusion, bicarbonate secretion, and chloride absorption depending on the specific subtype. The activity of H(+)-ATPases is regulated by acid-base status and several hormones, including angiotensin II and aldosterone. Angiotensin II stimulates chloride absorption mediated by pendrin in type B intercalated cells and this process is energized by the activity of H(+)-ATPases. Moreover, angiotensin II stimulates bicarbonate secretion by the connecting tubule (CNT) and early cortical collecting duct (CCD). In the present study we examined the effect of angiotensin II (10 nM) on H(+)-ATPase activity and localization in isolated mouse connecting tubules and cortical collecting ducts. Angiotensin II stimulated Na(+)-independent intracellular pH recovery about 2-3 fold, and this was abolished by the specific H(+)-ATPase inhibitor concanamycin. The effect of angiotensin II was mediated through type 1 angiotensin II receptors (AT(1)-receptors) because it could be blocked by saralasin. Stimulation of H(+)-ATPase activity required an intact microtubular network--it was completely inhibited by colchicine. Immunocytochemistry of isolated CNT/CCDs incubated in vitro with angiotensin II suggests enhanced membrane associated staining of H(+)-ATPases in pendrin expressing intercalated cells. In summary, angiotensin II stimulates H(+)-ATPases in CNT/CCD intercalated cells, and may contribute to the regulation of chloride absorption and bicarbonate secretion in this nephron segment.


Tomatidine Improves Pulmonary Inflammation in Mice with Acute Lung Injury.
Tomatidine, which is isolated from green tomato, can ameliorate inflammation and oxidative stress in cells and animal experiments and has been shown to improve airway inflammation in a murine model of asthma. Here, we investigated whether tomatidine can ameliorate acute lung injury in mice. Mice were given tomatidine by intraperitoneal injection for 7 consecutive days, and then, lung injury was induced via intratracheal instillation of lipopolysaccharide (LPS). Tomatidine reduced inflammatory cytokine expressions in bronchoalveolar lavage fluid (BALF), attenuated neutrophil infiltration in the BALF and lung tissue, increased superoxide dismutase activity and glutathione levels, and alleviated myeloperoxidase expression in the lung tissue of mice with lung injury. Tomatidine also decreased inflammatory cytokine and chemokine gene expression in inflammatory lungs and attenuated the phosphorylation of mitogen-activated protein kinase and nuclear factor kappa B. Furthermore, tomatidine enhanced the production of heme oxygenase-1, decreased the secretion of inflammatory cytokines and chemokines in LPS-stimulated lung epithelial cells, and attenuated THP-1 monocyte adhesion. Our findings suggest that tomatidine attenuates oxidative stress and inflammation, improving acute lung injury in mice.


Differences in videofluoroscopic swallowing study (VFSS) findings according to the vascular territory involved in stroke.
Dysphagia affects up to half of stroke patients and increases the risk of pneumonia and fatal outcomes. In order to assess swallowing difficulty, videofluoroscopic swallowing study (VFSS) has traditionally been the gold standard. The purpose of this study was to compare the patterns of post-stroke swallowing difficulties according to the vascular territories involved in the stroke. One hundred and three patients who were diagnosed with first ischemic stroke by brain magnetic resonance imaging and had swallowing difficulty were included in this study. Location of the stroke was classified into three groups: territorial anterior infarcts (TAI) (n = 62), territorial posterior infarcts (TPI) (n = 19) and white matter disease (WMD) (n = 22). Oral cavity residue existed significantly in the TAI group more than in any other groups (P = 0.017). The WMD group showed more residue in the valleculae (P = 0.002) and the TPI group showed more residue in the pyriform sinuses (P = 0.001). The oral transit time, pharyngeal delay time and pharyngeal transit time did not show significant differences among the groups with swallowing of both thick and thin liquids. Penetration and aspiration were more frequent in the TPI group (P < 0.05) with swallowing of both thick and thin liquids. The results suggest that TAI is more related to oral phase dysfunction and TPI is more related to pharyngeal dysfunction. In ischemic stroke, patterns of swallowing difficulty may differ according to the vascular territory involved and this should be considered in the management of post-stroke dysphagia.


Milrinone modulates endotoxemia, systemic inflammation, and subsequent acute phase response after cardiopulmonary bypass (CPB).
Compromised splanchnic perfusion and the resulting intestinal mucosal injury leads to a decreased mucosal barrier function, which allows translocation of intestinal flora and endotoxemia. The authors evaluated the effects of milrinone on splanchnic oxygenation, systemic inflammation, and the subsequent acute-phase response in patients undergoing coronary artery bypass grafting. This open, placebo-controlled randomized clinical study enrolled 22 adult patients in two groups. Before induction of anesthesia, baseline values were obtained and patients were randomized to receive milrinone (30 microg/kg bolus administered progressively in 10 min, followed by a continuous infusion of 0.5 microg x kg(-1) x min(-1)) or saline. The following parameters were determined: hemodynamics; systemic oxygen delivery and uptake; arterial, mixed venous and hepatic venous oxygen saturation; intramucosal pH (pHi); and mixed and hepatic venous plasma concentrations of endotoxin, interleukin 6, serum amyloid A, and C-reactive protein. Milrinone did not prevent gastrointestinal acidosis as measured by pHi, but its perioperative administration resulted in significantly higher pHi levels compared with control. Venous and hepatic venous endotoxin and the interleukin 6 concentration were reduced significantly in the milrinone group. Serum amyloid A values were attenuated in the milrinone group 24 h after surgery. No significant differences could be seen in routinely measured oxygen transport-derived variables. Perioperative administration of low-dose milrinone may have antiinflammatory properties and may improve splanchnic perfusion in otherwise healthy patients undergoing routine coronary artery bypass grafting.


Kinesthetic Feedback During 2DOF Wrist Movements via a Novel MR-Compatible Robot.
We demonstrate the interaction control capabilities of the MR-SoftWrist, a novel MR-compatible robot capable of applying accurate kinesthetic feedback to wrist pointing movements executed during fMRI. The MR-SoftWrist, based on a novel design that combines parallel piezoelectric actuation with compliant force feedback, is capable of delivering 1.5 N [Formula: see text] of torque to the wrist of an interacting subject about the flexion/extension and radial/ulnar deviation axes. The robot workspace, defined by admissible wrist rotation angles, fully includes a circle with a 20 deg radius. Via dynamic characterization, we demonstrate capability for transparent operation with low (10% of maximum torque output) backdrivability torques at nominal speeds. Moreover, we demonstrate a 5.5 Hz stiffness control bandwidth for a 14 dB range of virtual stiffness values, corresponding to 25%-125% of the device's physical reflected stiffness in the nominal configuration. We finally validate the possibility of operation during fMRI via a case study involving one healthy subject. Our validation experiment demonstrates the capability of the device to apply kinesthetic feedback to elicit distinguishable kinetic and neural responses without significant degradation of image quality or task-induced head movements. With this study, we demonstrate the feasibility of MR-compatible devices like the MR-SoftWrist to be used in support of motor control experiments investigating wrist pointing under robot-applied force fields. Such future studies may elucidate fundamental neural mechanisms enabling robot-assisted motor skill learning, which is crucial for robot-aided neurorehabilitation.


The genetic prehistory of the New World Arctic.
The New World Arctic, the last region of the Americas to be populated by humans, has a relatively well-researched archaeology, but an understanding of its genetic history is lacking. We present genome-wide sequence data from ancient and present-day humans from Greenland, Arctic Canada, Alaska, Aleutian Islands, and Siberia. We show that Paleo-Eskimos (~3000 BCE to 1300 CE) represent a migration pulse into the Americas independent of both Native American and Inuit expansions. Furthermore, the genetic continuity characterizing the Paleo-Eskimo period was interrupted by the arrival of a new population, representing the ancestors of present-day Inuit, with evidence of past gene flow between these lineages. Despite periodic abandonment of major Arctic regions, a single Paleo-Eskimo metapopulation likely survived in near-isolation for more than 4000 years, only to vanish around 700 years ago.


Development of a genetic tool for product regulation in the diverse British pig breed market.
The application of DNA markers for the identification of biological samples from both human and non-human species is widespread and includes use in food authentication. In the food industry the financial incentive to substituting the true name of a food product with a higher value alternative is driving food fraud. This applies to British pork products where products derived from traditional pig breeds are of premium value. The objective of this study was to develop a genetic assay for regulatory authentication of traditional pig breed-labelled products in the porcine food industry in the United Kingdom. The dataset comprised of a comprehensive coverage of breed types present in Britain: 460 individuals from 7 traditional breeds, 5 commercial purebreds, 1 imported European breed and 1 imported Asian breed were genotyped using the PorcineSNP60 beadchip. Following breed-informative SNP selection, assignment power was calculated for increasing SNP panel size. A 96-plex assay created using the most informative SNPs revealed remarkably high genetic differentiation between the British pig breeds, with an average FST of 0.54 and Bayesian clustering analysis also indicated that they were distinct homogenous populations. The posterior probability of assignment of any individual of a presumed origin actually originating from that breed given an alternative breed origin was > 99.5% in 174 out of 182 contrasts, at a test value of log(LR) > 0. Validation of the 96-plex assay using independent test samples of known origin was successful; a subsequent survey of market samples revealed a high level of breed label conformity. The newly created 96-plex assay using selected markers from the PorcineSNP60 beadchip enables powerful assignment of samples to traditional breed origin and can effectively identify mislabelling, providing a highly effective tool for DNA analysis in food forensics.


Variance in the reproductive success of flat oyster Ostrea edulis L. assessed by parentage analyses in natural and experimental conditions.
In order to document further the phenomena of variance in reproductive success in natural populations of the European flat oyster Ostrea edulis, two complementary studies based on natural and experimental populations were conducted. The first part of this work was focused on paternity analyses using a set of four microsatellite markers for larvae collected from 13 brooding females sampled in Quiberon Bay (Brittany, France). The number of individuals contributing as the male parent to each progeny assay was highly variable, ranging from 2 to more than 40. Moreover, paternal contributions showed a much skewed distribution, with some males contributing to 50-100% of the progeny assay. The second part of this work consisted of the analysis of six successive cohorts experimentally produced from an acclimated broodstock (62 wild oysters sampled in the Quiberon Bay). Allelic richness was significantly higher in the adult population than in the temporal cohorts collected. Genetic differentiation (F(st) estimates) was computed for each pair of samples and all significant values ranged from 0.7 to 11.9%. A limited effective number of breeders (generally below 25) was estimated in the six temporal cohorts. The study gives first indications of the high variance in reproductive success as well as a reduced effective size, not only under experimental conditions but also in the wild. Surprisingly, the pool of the successive cohorts, based on the low number of loci used, appeared to depict a random and representative set of alleles of the progenitor population, indicating that the detection of patterns of temporal genetic differentiation at a local scale most likely depends on the sampling window.


Phosphatidylserine exposure on the surface of Leishmania amazonensis amastigotes modulates in vivo infection and dendritic cell function.
Leishmania amazonensis parasites can cause diverse forms of leishmaniasis in humans and persistent lesions in most inbred strains of mice. In both cases, the infection is characterized by a marked immunosuppression of the host. We previously showed that amastigote forms of the parasite make use of surface-exposed phosphatidylserine (PS) molecules to infect host cells and promote alternative macrophage activation, leading to uncontrolled intracellular proliferation of the parasites. In this study, we demonstrated that treatment of infected mice with a PS-targeting monoclonal antibody ameliorated parasite loads and lesion development, which correlated with increased proliferative responses by lymphocytes. In addition, we observed an enhanced dendritic cell (DC) activation and antigen presentation in vitro. Our data imply that the recognition of PS exposed on the surface of amastigotes plays a role in down-modulating DC functions, in a matter similar to that of apoptotic cell clearance. This study provides new information regarding the mechanism of immune suppression in Leishmania infection.


Facile fabrication of high-efficiency near-infrared absorption film with tungsten bronze nanoparticle dense layer.
An excellent transparent film with effective absorption property in near-infrared (NIR) region based on cesium-doped tungsten oxide nanoparticles was fabricated using a facile double layer coating method via the theoretical considerations. The optical performance was evaluated; the double layer-coated film exhibited 10% transmittance at 1,000 nm in the NIR region and over 80% transmittance at 550 nm in the visible region. To optimize the selectivity, the optical spectrum of this film was correlated with a theoretical model by combining the contributions of the Mie-Gans absorption-based localized surface plasmon resonance and reflections by the interfaces of the heterogeneous layers and the nanoparticles in the film. Through comparison of the composite and double layer coating method, the difference of the nanoscale distances between nanoparticles in each layer was significantly revealed. It is worth noting that the nanodistance between the nanoparticles decreased in the double layer film, which enhanced the optical properties of the film, yielding a haze value of 1% or less without any additional process. These results are very attractive for the nanocomposite coating process, which would lead to industrial fields of NIR shielding and thermo-medical applications. 78.67.Sc; 78.67.Bf; 81.15.-z.


Epigenetic characterization of the CpG islands of bovine Leptin and POU5F1 genes in cloned bovine fetuses.
Abnormal development and fetal loss during postimplantation period are concerns for production of nuclear transferred animals. Aberrant DNA methylation is one of the reasons for poor survival of cloned animals. In mammalian genome DNA, CpG islands are preferentially located at the start of transcription of housekeeping genes and are associated with tissue-specific genes. The correct and consecutive mechanisms of DNA methylation in the CpG islands are necessary for selective gene expressions that determine the properties of individual cells, tissues, and organs. In this study, we investigated the methylation status of the CpG islands of the bovine Leptin and POU5F1 genes in fetal and placental tissues from fetuses produced by artificial insemination (AI) and nuclear transfer (NT) at days 48 and 59 of pregnancy. Altered DNA methylation was observed in the normal and cloned fetal, placental, and endometrial tissues using bisulfite sequencing and pyrosequencing. Different tissue-specific methylated regions in the bovine Leptin and POU5F1 genes show a variable methylation status in NT fetuses compared to AI control.


Determinants of the progression in lumbar degeneration: a 5-year follow-up study of adult male monozygotic twins.
A 5-year follow-up study of exposure of discordant monozygotic twin pairs with repeated interviews and spine imaging. The primary goals were to record changes in the degenerative signs over a 5-year interval and to estimate the effects of familial influences and suspected environmental risk factors on the speed of lumbar degeneration. Traditionally, disc degeneration has been attributed to aging and environmental exposures; recently, a dominant effect of genetics has been revealed. Yet the etiopathogenesis of disc degeneration remains poorly understood and controversial despite being a primary target of diagnostic and therapeutic interventions. Among 116 monozygotic twin pairs, which had been examined 5 years earlier, 75 pairs (150 men) were reexamined. They were imaged using the same MRI scanner and examination protocol as at baseline. The data were analyzed using statistical methods for longitudinal studies. Progression in disc height narrowing, disc bulging, osteophytosis, and fatty degeneration in the lumbar spine was seen in about 7% to 13% of the discs in 7% to 46% of subjects during 5-year follow-up. Few degenerative findings appear to reverse; few disc height measures increased, some anular tears were no longer visible, and bulging/herniation diminished. New anular tears (in axial view) were detected in 1.5%, disappeared in 2%, and were unchanged in 5.3% of discs; in the sagittal view, new high intensity zones findings were identified in 0.5%, were no longer apparent in 1.6%, and were unchanged in 7.1% of discs. There were no clear changes in upper endplates: in 2.1% of discs, the irregularity score increased and in 1.8% it decreased. Familial aggregation, reflecting genetic, and shared environmental influences, explained 47% to 66% of the variance in progression of degenerative signs on lumbar MRI, and resistance training and occupational physical loading together explained 2% to 10% of the progression in the degenerative signs in lumbar MRIs. Progression of disc height narrowing, bulging, osteophytes, and fatty degeneration was detected in about 10% or less of the T12-S1 discs. Development and disappearance of anular lesions were rarer. No clear changes were seen in endplate irregularities. The results also confirm that hereditary effects have a dominant role in the progression of disc degeneration and suggest that occupational lifting and leisure time resistance training have modest additional effects.


Assessment of fatigue in routine care on a Multidimensional Health Assessment Questionnaire (MDHAQ): a cross-sectional study of associations with RAPID3 and other variables in different rheumatic diseases.
To characterise associations of fatigue with other variables within a multidimensional health assessment questionnaire (MDHAQ) in routine care of patients with different rheumatic diagnoses. All patients complete MDHAQ, which includes fatigue on a 0-10 visual analogue scale (VAS), and routine assessment of patient index data (RAPID3), a composite of function, pain, and patient global. Physicians complete a RheuMetric checklist which includes 4 VAS for overall global status (DOCGL), inflammation, damage, and distress. Median score for fatigue and other MDHAQ and RheuMetric scores were compared in 4 diagnosis groups: rheumatoid arthritis (RA), osteoarthritis (OA), systemic lupus erythematosus (SLE), and fibromyalgia (FM), using a Kruskall-Wallis test. Associations of fatigue with other variables were analysed using Spearman correlations and multivariate regressions. 612 patients were included: 173 RA, 199 with OA, 146 with SLE, and 94 with FM. Median fatigue was significantly higher in FM (7) than in RA (4), OA (5), and SLE (5). Fatigue was correlated significantly with all other MDHAQ scores, at higher levels in RA and SLE versus OA and FM. Fatigue was correlated significantly with DOCGL in RA, OA, SLE, but not FM. In multivariate analyses, fatigue scores were explained independently by higher pain and symptom number in RA; lower age and higher symptom number in OA; only higher pain in SLE; and none of the variables in FM. Fatigue is common in rheumatic diseases and strongly associated with higher pain and number of symptoms. The MDHAQ provides a useful tool to assess fatigue in clinical settings.