Genome-wide diversity and demographic dynamics of Cameroon goats and their divergence from east African, north African, and Asian conspecifics.
Indigenous goats make significant contributions to Cameroon's national and local economy, but little effort has been devoted to identifying the populations. Here, we assessed the genetic diversity and demographic dynamics of Cameroon goat populations using mitochondrial DNA (two populations) and autosomal markers (four populations) generated with the Caprine 50K SNP chip. To infer genetic relationships at continental and global level, genotype data on six goat populations from Ethiopia and one population each from Egypt, Morocco, Iran, and China were included in the analysis. The mtDNA analysis revealed 83 haplotypes, all belonging to haplogroup A, in Cameroon goats. Four haplotypes were shared between goats found in Cameroon, Mozambique, Namibia, Zimbabwe, Kenya, and Ethiopia. Analysis of autosomal SNPs in Cameroon goats revealed the lowest HO (0.335±0.13) and HE (0.352±0.15) in the North-west Highland and Central Highland populations, respectively. Overall, the highest HO (0.401±0.12) and HE (0.422±0.12) were found for Barki and Iranian goats, respectively. Barki goats had the highest average MAF, while Central Highland Cameroon goats had the lowest. Overall, Cameroon goats demonstrated high FIS. AMOVA revealed that 13.29% of the variation was explained by genetic differences between the six population groups. Low average FST (0.01) suggests intermixing among Cameroon goats. All measures indicated that Cameroon goats are closer to Moroccan goats than to other goat populations. PCA and STRUCTURE analyses poorly differentiated the Cameroon goats, as did genetic distance, Neighbor-Net network, and neighbor-joining tree analyses. The haplotype analysis of mtDNA showed the initial dispersion of goats to Cameroon and central Africa from north-east Africa following the Nile Delta. Whereas, the approximate Bayesian computation indicated Cameroon goats were separated from Moroccan goats after 506 generations in later times (~1518 YA), as supported by the phylogenetic net-work and admixture outputs. Overall, indigenous goats in Cameroon show weak phylogenetic structure, suggesting either extensive intermixing.


Astigmatic change in biaxial microincisional cataract surgery with enlargement of one incision: a prospective controlled study.
To investigate the astigmatic changes induced by biaxial microincisional cataract surgery with enlargement of one incision to accommodate a foldable standard lens. In a prospective controlled trial one eye each of 50 patients was evaluated. Two temporal microincisions were applied and the right-hand incision was enlarged to 2.8 mm to allow for the insertion of the lens. Based on preoperative and postoperative keratometry readings, vector analysis was performed to calculate the surgically induced astigmatism (SIA) and the flattening effect of each of the two incisions at their meridia. To assess the degree of measurement error, a non-surgical control group underwent keratometry over the same time frame and vector analysis was used to calculate the same indices based on the variation in the keratometry readings. Six weeks postoperatively, mean SIA was 0.66 +/- 0.08 D in the study group and 0.51 +/- 0.05 D in the non-surgical control group (P = 0.041). At the right-hand incision, a flattening of 0.34 +/- 0.09 D was measured in the study group as opposed to a steepening of 0.03 +/- 0.06 D in the control group (P < 0.001). At the smaller left-hand incision, steepening effects of 0.28 +/- 0.08 and 0.05 +/- 0.06 D occurred, respectively (P = 0.011). Biaxial microincisional cataract surgery with enlargement of one incision to 2.8 mm is not astigmatically neutral, demonstrating a statistically significantly larger SIA than that attributable to measurement error.


Haze activity of different barley trypsin inhibitors of the chloroform/methanol type (BTI-CMe).
Our previous study found that the critical protein in SE (silica eluted) proteins is BTI-CMe, and assumed that SE-ve malt for brewing may improve the haze stability in beer. In this study, we investigated the difference in gene sequence and corresponding amino acid sequence of BTI-CMe between SE+ve and SE-ve types. The results showed that there were 7 amino acid differences between Yerong (SE-ve) and Franklin (SE+ve). Two types BTI-CMe were expressed in vitro and purified successfully. By adding the purified BTI-CMe into commercial beer, we found that both original turbidity and alcohol chill haze degree of beer were increased. BTI-CMe of SE-ve haplotype showed a lower level of haze formation in beer than SE+ve haplotype. Response surface methodology (RSM) was conducted to determine the relationship between BTI-CMe and tannic acid, and their effects on haze formation. It was found that (1) higher content of BTI-CMe and/or tannic acid in beer would give rise to higher turbidity; (2) there was a significant interaction between BTI-CMe and tannic acid; (3) haze activity disparity of BTI-CMe between two types was significantly and positively correlated with the tannic acid concentration.


Global aspects of pacC regulation of pathogenicity genes in Colletotrichum gloeosporioides as revealed by transcriptome analysis.
Colletotrichum gloeosporioides alkalinizes its surroundings during colonization of host tissue. The transcription factor pacC is a regulator of pH-controlled genes and is essential for successful colonization. We present here the sequence assembly of the Colletotrichum fruit pathogen and use it to explore the global regulation of pathogenicity by ambient pH. The assembled genome size was 54 Mb, encoding 18,456 genes. Transcriptomes of the wild type and ΔpacC mutant were established by RNA-seq and explored for their global pH-dependent gene regulation. The analysis showed that pacC upregulates 478 genes and downregulates 483 genes, comprising 5% of the fungal genome, including transporters, antioxidants, and cell-wall-degrading enzymes. Interestingly, gene families with similar functionality are both up- and downregulated by pacC. Global analysis of secreted genes showed significant pacC activation of degradative enzymes at alkaline pH and during fruit infection. Select genes from alkalizing-type pathogen C. gloeosporioides and from acidifying-type pathogen Sclerotinia sclerotiorum were verified by quantitative reverse-transcription polymerase chain reaction analysis at different pH values. Knock out of several pacC-activated genes confirmed their involvement in pathogenic colonization of alkalinized surroundings. The results suggest a global regulation by pacC of key pathogenicity genes during pH change in alkalinizing and acidifying pathogens.


The Brain-Heart Connection in Sympathetically Triggered Inherited Arrhythmia Syndromes.
Sympathetically triggered inherited arrhythmia syndromes, including the long QT syndrome (LQTS) and catecholaminergic polymorphic ventricular tachycardia (CPVT), can cause sudden cardiac death in young individuals with structurally normal hearts. With cardiac events typically triggered by physical or emotional stress, not surprisingly, two of the most common treatments are neuromodulators, including mainstay beta blocker pharmacotherapy, and surgical sympathetic cardiac denervation. This review updates the clinician on the relevant anatomy and physiology of the cardiac autonomic nervous system, outlines neurocardiac arrhythmia mechanisms, and discusses the latest rationale for a neurocardiac therapeutic approach to manage sympathetic-induced arrhythmia in patients with inherited cardiac disease.


Smad4 inactivation predicts for worse prognosis and response to fluorouracil-based treatment in colorectal cancer.
To determine whether expression of Smad4, a tumour suppressor found to be absent in 10% of colorectal cancer (CRC), is associated with outcomes in patients with CRC. Tumour samples from 241 consecutive patients with CRC who underwent upfront colon resection between 2005 and 2009 were obtained. Triplicate tissue cores from resected primary colon tumours and matched normal controls were used to construct the tissue microarrays (TMAs). We examined the expression of Smad4 using immunohistochemistry. Clinicopathological records were obtained for all patients. TMAs were reviewed by two pathologists and scored as either 'positive' or 'negative' for nuclear staining. In total, 21 of 241 tumours (8.6%) were Smad4 negative. Loss of Smad4 expression correlated with significantly worse overall survival (OS) (p=0.011) and disease-free survival (DFS) (p=0.024). Patients with loss of Smad4 expression had a median OS of 31 months compared with 89 months positive Smad4 expression. Loss of Smad4 remained significant on multivariate analysis for OS (p=0.0097). In patients with node-positive disease, loss of Smad4 predicts for worse DFS (p=0.012). In patients with metastatic and recurrent disease, Smad4 loss predicts for worse OS (p=0.012). Of the patients that received capecitabine over the course of their treatment, those with Smad4 loss (n=13) had significantly worse DFS (p=0.003) and OS (p=0.0007). Loss of Smad4 expression is associated with worse DFS and OS in multiple subsets of patients with CRC. Further studies are required to validate our findings and ascertain the role of Smad4 status in the management of this disease.


[Meibomian gland dysfunction].
Meibomian gland dysfunction (MGD) is a chronic disease, usually caused by obstruction of the secretory Meibomian glands. The subsequent reduction of gland secretion results in a decreased amount of lipids in the tear film. This results in a faster evaporation of the tear film and thus an evaporative dry eye. MGD alone is responsible for about 60% of all cases in combination with aqueous deficiency for a further 20% of dry eyes. While in Europe up to 20% of the population are suffering from MDD, this is true in Asia for over 60% of the population. MGD is more common in women and it incidence increases with age. It is influenced by the hormonal status as well as chemical and mechanical noxious stimuli. Additional risk factors include various skin diseases such as rosacea, acne or atopy. To diagnose MGD, particular attention should be paid to changes in the lid margin such as plugging or pouting of the ducts, thickening and telangiectasia. However, most important is the diagnostic expression of the glands. At first it should be assessed whether secretion can be caused by pressure to the eyelid against the globe and secondly the quality of the expressed secretions should be evaluated. MGD should be treated according to the severity of the disease. While in mild stages instructions for lid margin hygiene, warming and massage in combination with artificial tears might be sufficient, in more severe stages oral tetracyclin derivatives and anti-inflammatory eye drops such as steroids or CSA are necessary for successful treatment.


Recovery of function after serial ablation of prefrontal cortex in the rhesus monkey.
Rhesus monkeys with one-stage or serial ablation of sulcus principalis (prefrontal association cortex) were compared on three spatial tasks. On all tests, the serial monkeys made fewer errors than did the monkeys with onestage lesions. These results indicate that partial recovery of function can occur after extensive destruction of association cortex in the mature primate brain if the damage is distributed over a number of operations.


Development and validation of the Lesion Synthesis Toolbox and the Perception Study Tool for quantifying observer limits of detection of lesions in positron emission tomography.
Purpose: Accurate detection of cancer lesions in positron emission tomography (PET) is fundamental to achieving favorable clinical outcomes. Therefore, image reconstruction, processing, visualization, and interpretation techniques must be optimized for this task. The objective of this work was to (1) develop and validate an efficient method to generate well-characterized synthetic lesions in real patient data and (2) to apply these lesions in a human perception experiment to establish baseline measurements of the limits of lesion detection as a function of lesion size and contrast using current imaging technologies. Approach: A fully integrated software package for synthesizing well-characterized lesions in real patient PET was developed using a vendor provided PET image reconstruction toolbox (REGRECON5, General Electric Healthcare, Waukesha, Wisconsin). Lesion characteristics were validated experimentally for geometric accuracy, activity accuracy, and absence of artifacts. The Lesion Synthesis Toolbox was used to generate a library of 133 synthetic lesions of varying sizes ( n = 7 ) and contrast levels ( n = 19 ) in manually defined locations in the livers of 37 patient studies. A lesion-localization perception study was performed with seven observers to determine the limits of detection with regard to lesion size and contrast using our web-based perception study tool. Results: The Lesion Synthesis Toolbox was validated for accurate lesion placement and size. Lesion intensities were deemed accurate with slightly elevated activities (5% at 2:1 lesion-to-background contrast) in small lesions ( Ø = 15 mm spheres), and no bias in large lesions ( Ø = 22.5 mm ). Bed-stitching artifacts were not observed, and lesion attenuation correction bias was small ( - 1.6 ± 1.2 % ). The 133 liver lesions were synthesized in ∼ 50 h , and readers were able to complete the perception study of these lesions in 12 ± 3 min with consistent limits of detection amongst all readers. Conclusions: Our open-source utilities can be employed by nonexperts to generate well-characterized synthetic lesions in real patient PET images and for administering perception studies on clinical workstations without the need to install proprietary software.


Prenatal stress potentiates pilocarpine-induced epileptic behaviors in infant rats both time and sex dependently.
Stressful events during gestation have important effects on the later physical and mental health of the offspring. In the study described here, the pilocarpine-induced seizure model was used to test the hypothesis that prenatal stress affects seizure susceptibility in infant rats. Prenatal stress consisted of daily restraint of the dam under normal room conditions (for 120minutes, twice daily) during the first, second, and third weeks of gestation. The pups were then compared with pups born to unstressed dams. Both second- and third-week-gestation stress significantly reduced pilocarpine-induced seizures in 19-day-old rat offspring, as compared with nonstressed control offspring. Mid- and late-gestation stress increased the rate and time of tonic-clonic seizures. Mortality rate 2 and 24hours after pilocarpine administration increased significantly in all stressed rats. Stress induced a significant rise in circulating corticosterone levels (2- to 8-fold, P<0.001) in the offspring. Female offspring differed little from male offspring with respect to blood corticosterone levels and epileptic behaviors. These findings indicate that prenatal stress, particularly during the second and third weeks of pregnancy, may play an important role in increasing seizure vulnerability in the unborn offspring. Female rats are more resistant to stress than males probably because of the lower susceptibility of their hypothalamic-pituitary-adrenal axis.


Niacin ameliorates oxidative stress, inflammation, proteinuria, and hypertension in rats with chronic renal failure.
Significant reduction of renal mass causes progressive deterioration of renal function and structure which is mediated by systemic and glomerular hypertension, hyperfiltration, oxidative stress, inflammation, and dyslipidemia. Niacin is known to improve lipid metabolism and exert antioxidant/anti-inflammatory actions. Therefore, we considered that niacin supplementation may attenuate oxidative stress, inflammation, and tissue injury in the remnant kidney. To this end, 56 nephrectomized [chronic kidney disease (CKD)] rats were randomly assigned to niacin-treated (50 mg x kg(-1) x day(-1) in the drinking water for 12 wk) and untreated groups. Sham-operated rats served as controls. The untreated CKD rats exhibited azotemia, hypertension, hypertriglyceridemia, proteinuria, glomerulosclerosis, tubulointerstitial damage, upregulation of MCP-1, plasminogen activator inhibitor-1 (PAI-1), transforming growth factor (TGF)-beta, cyclooxygenase (COX)-1, COX-2, and NAD(P)H oxidase (NOX-4, gp91(phox), p47(phox) and p22(phox) subunits) and activation of NF-kappaB (IkappaB phosphorylation). Niacin administration reduced MCP-1, PAI-1, TGF-beta, p47(phox), p22(phox), COX-1, and NF-kappaB activation, ameliorated hypertension, proteinuria, glomerulosclerosis, and tubulointerstitial injury. Although niacin lowered serum creatinine and raised creatinine clearance, the differences did not reach statistical significance. Thus niacin supplementation helps to attenuate histological injury and mitigate upregulation of oxidative and inflammatory systems in the remnant kidney.


Ground Reaction Forces Generated by Twenty-eight Hatha Yoga Postures.
Adherents claim many benefits from the practice of yoga, including promotion of bone health and prevention of osteoporosis. However, no known studies have investigated whether yoga enhances bone mineral density. Furthermore, none have estimated reaction forces applied by yoga practitioners. The purpose of this study was to collect ground reaction force (GRF) data on a variety of hatha yoga postures that would commonly be practiced in fitness centers or private studios. Twelve female and eight male volunteers performed a sequence of 28 hatha yoga postures while GRF data were collected with an AMTI strain-gauge force platform. The sequence was repeated six times by each study subject. Four dependent variables were studied: peak vertical GRF, mean vertical GRF, peak resultant GRF, and mean resultant GRF. Univariate analysis was used to identify mean values and standard deviations for the dependent variables. Peak vertical and resultant values of each posture were similar for all subjects, and standard deviations were small. Similarly, mean vertical and resultant values were similar for all subjects. This 28 posture yoga sequence produced low impact GRF applied to upper and lower extremities. Further research is warranted to determine whether these forces are sufficient to promote osteogenesis or maintain current bone health in yoga practitioners.


Psychophysics of inspiratory muscle force.
The perceived magnitude of static inspiratory muscle pressure was studied in normal subjects using psychophysical techniques. The sensory magnitude of a range of inspiratory pressures increased as the magnitude of the pressure increased. When the duration of the inspiratory pressure was controlled, the sensory magnitude also increased as duration increased. The relationship can be described by a single psychophysical function, psi = k x P1.234 x t0.62, where psi is perceived magnitude, P is inspiratory pressure, t is duration, and k is a constant. Use of different muscle groups and changes in lung volume altered the perceived magnitude of static inspiratory pressures. When static inspiratory pressures were generated by the abdomen-diaphragm, the perceived magnitude was significantly greater (P less than 0.01) than when they were generated by the rib cage. When lung volume was increased, the perceived magnitude of pressure was reduced. The results show that the perceived magnitude of static inspiratory pressures is affected by the pressure itself, pressure duration, the muscles used, and the lung volume at which the pressure is generated.


The insulin-like growth factor system in normal mammary gland function.
Insulin-like growth factors (IGF) are now known to play an important role in normal mammary gland development and have been implicated as risk factors in the etiology of breast cancer. Studies in genetically engineered mouse models have demonstrated that the IGF system acts within the mammary epithelium at key developmental stages to either stimulate cell cycle progression or inhibit cell death. The importance of these two actions to mammary gland development varies with the different developmental stage. Cell cycle effects appear to predominate during virgin ductal development and early pregnancy while cell survival effects are important for initiation and maintenance of lactation. This variation with stage of development most likely reflects changes in specific populations of epithelial cells within the gland. Relatively little information exists however on the actions of the IGF system during fetal or very early postnatal development. In addition much remains to be learned about the intracellular signaling mechanism that mediate the effects of IGFs on the different populations of epithelial cells present within the gland. This review summarizes the current state of knowledge concerning IGF actions, and mechanisms of action, within the normal mammary gland.


Patient safety education at Japanese nursing schools: results of a nationwide survey.
Patient safety education is becoming of worldwide interest and concern in the field of healthcare, particularly in the field of nursing. However, as elsewhere, little is known about the extent to which nursing schools have adopted patient safety education into their curricula. We conducted a nationwide survey to characterize patient safety education at nursing schools in Japan. Response rate was 43% overall. Ninety percent of nursing schools have integrated the topic of patient safety education into their curricula. However, 30% reported devoting less than five hours to the topic. All schools use lecture based teaching methods while few used others, such as role playing. Topics related to medical error theory are widely taught, e.g. human factors and theories & models (Swiss Cheese Model, Heinrich's Law) while relatively few schools cover practical topics related to error analysis such as root cause analysis. Most nursing schools in Japan cover the topic of patient safety, but the number of hours devoted is modest and teaching methods are suboptimal. Even so, national inclusion of patient safety education is a worthy, achievable goal.


Exploiting heterogeneous environments: does photosynthetic acclimation optimize carbon gain in fluctuating light?
Plants have evolved complex mechanisms to balance the efficient use of absorbed light energy in photosynthesis with the capacity to use that energy in assimilation, so avoiding potential damage from excess light. This is particularly important under natural light, which can vary according to weather, solar movement and canopy movement. Photosynthetic acclimation is the means by which plants alter their leaf composition and structure over time to enhance photosynthetic efficiency and productivity. However there is no empirical or theoretical basis for understanding how leaves track historic light levels to determine acclimation status, or whether they do this accurately. We hypothesized that in fluctuating light (varying in both intensity and frequency), the light-response characteristics of a leaf should adjust (dynamically acclimate) to maximize daily carbon gain. Using a framework of mathematical modelling based on light-response curves, we have analysed carbon-gain dynamics under various light patterns. The objective was to develop new tools to quantify the precision with which photosynthesis acclimates according to the environment in which plants exist and to test this tool on existing data. We found an inverse relationship between the optimal maximum photosynthetic capacity and the frequency of low to high light transitions. Using experimental data from the literature we were able to show that the observed patterns for acclimation were consistent with a strategy towards maximizing daily carbon gain. Refinement of the model will further determine the precision of acclimation.


Effects of Caffeine and Acute Aerobic Exercise on Working Memory and Caffeine Withdrawal.
Studies show that a single bout of exercise confers cognitive benefits. However, many individuals use psychoactive substances such as caffeine to enhance cognitive performance. The effects of acute exercise in comparison to caffeine on cognition remain unknown. Furthermore, caffeine use is associated with withdrawal symptoms upon cessation. Whether acute exercise can reduce withdrawal symptoms also remains unknown. The objectives of this study were to compare the effects of acute moderate intensity aerobic exercise to caffeine on working memory (WM) and caffeine withdrawal symptoms (CWS). In Phase I, non-caffeine (n = 29) and caffeine consumers (n = 30) completed a WM assessment, followed by acute exercise and caffeine. In Phase II, caffeine consumers (n = 25) from Phase I underwent the WM assessment and reported CWS following a 12-hour deprivation period. Acute moderate intensity aerobic exercise and caffeine (1.2 mg/kg) significantly improved WM accuracy and reduced CWS comparably. WM performance was not reduced following caffeine deprivation.


Production of Recombinant Peanut Allergen Ara h 2 using Lactococcus lactis.
Natural allergen sources can supply large quantities of authentic allergen mixtures for use as immunotherapeutics. However, such extracts are complex, difficult to define, vary from batch to batch, which may lead to unpredictable efficacy and/or unacceptable levels of side effects. The use of recombinant expression systems for allergen production can alleviate some of these issues. Several allergens have been tested in high-level expression systems and in most cases show immunereactivity comparable to their natural counterparts. The gram positive lactic acid bacterium Lactococcus lactis is an attractive microorganism for use in the production of protein therapeutics. L. lactis is considered food grade, free of endotoxins, and is able to secrete the heterologous product together with few other native proteins. Hypersensitivity to peanut represents a serious allergic problem. Some of the major allergens in peanut have been described. However, for therapeutic usage more information about the individual allergenic components is needed. In this paper we report recombinant production of the Ara h 2 peanut allergen using L. lactis. A synthetic ara h 2 gene was cloned into an L. lactis expression plasmid containing the P170 promoter and the SP310mut2 signal sequence. Flask cultures grown overnight showed secretion of the 17 kDa Ara h 2 protein. A batch fermentation resulted in 40 mg/L recombinant Ara h 2. Purification of Ara h 2 from the culture supernatant was done by hydrophobic exclusion and size separation. Mass spectrometry and N-terminal analysis showed a recombinant Ara h 2 of full length and correctly processed by the signal peptidase. The immunological activity of recombinant Ara h 2 was analysed by ELISA using antibodies specific for native Ara h 2. The recombinant Ara h 2 showed comparable immunereactivity to that of native Ara h 2. Recombinant production of Ara h 2 using L. lactis can offer high yields of secreted, full length and immunologically active allergen. The L. lactis expression system can support recombinant allergen material for immunotherapy and component resolved allergen diagnostics.


Neural Simpletrons: Learning in the Limit of Few Labels with Directed Generative Networks.
We explore classifier training for data sets with very few labels. We investigate this task using a neural network for nonnegative data. The network is derived from a hierarchical normalized Poisson mixture model with one observed and two hidden layers. With the single objective of likelihood optimization, both labeled and unlabeled data are naturally incorporated into learning. The neural activation and learning equations resulting from our derivation are concise and local. As a consequence, the network can be scaled using standard deep learning tools for parallelized GPU implementation. Using standard benchmarks for nonnegative data, such as text document representations, MNIST, and NIST SD19, we study the classification performance when very few labels are used for training. In different settings, the network's performance is compared to standard and recently suggested semisupervised classifiers. While other recent approaches are more competitive for many labels or fully labeled data sets, we find that the network studied here can be applied to numbers of few labels where no other system has been reported to operate so far.


Whole-Plant Metabolic Allocation Under Water Stress.
Trade-offs between plant growth and defense depend on environmental resource availability. Plants are predicted to prioritize growth when environmental resources are abundant and defense when environmental resources are scarce. Nevertheless, such predictions lack a whole-plant perspective-they do not account for potential differences in plant allocation above- and belowground. Such accounting is important because leaves and roots, though both critical to plant survival and fitness, differ in their resource-uptake roles and, often, in their vulnerability to herbivores. Here we aimed to determine how water availability affects plant allocation to multiple metabolic components of growth and defense in both leaves and roots. To do this, we conducted a meta-analysis of data from experimental studies in the literature. We assessed plant metabolic responses to experimentally reduced water availability, including changes in growth, nutrients, physical defenses, primary metabolites, hormones, and other secondary metabolites. Both above- and belowground, reduced water availability reduced plant biomass but increased the concentrations of primary metabolites and hormones. Importantly, however, reduced water had opposite effects in different organs on the concentrations of other secondary metabolites: reduced water increased carbon-based secondary metabolites in leaves but reduced them in roots. In addition, plants suffering from co-occurring drought and herbivory stresses exhibited dampened metabolic responses, suggesting a metabolic cost of multiple stresses. Our study highlights the needs for additional empirical studies of whole-plant metabolic responses under multiple stresses and for refinement of existing plant growth-defense theory in the context of whole plants.


A randomized double-blind placebo-controlled trial of cyclosporine in steroid-resistant idiopathic focal segmental glomerulosclerosis in children.
There is no generally accepted treatment for primary focal segmental glomerulosclerosis (FSGS). Steroids alone and steroids plus cyclophosphamide can be expected to induce a remission of the proteinuria in only 27% of patients. Probably the majority of FSGS patients will reach ESRD over the extended course of their disease. In addition to the work presented in this study, there have been many reports of the potential effectiveness of cyclosporine (CSA) on reducing the proteinuria of FSGS. This study was undertaken to test the efficacy and safety of a 6-month course of CSA in a double-blinded, prospectively randomized, placebo-controlled trial in children with corticosteroid-resistant FSGS. The potential inhibitory effect of hypercholesterolemia on the proteinuria-reducing actions of CSA was also assessed. Twenty-five patients with FSGS were randomized to receive either placebo or CSA for 6 months. Twelve of the 12 patients that received CSA experienced a diminution of their proteinuria as opposed to only two of the 12 placebo-treated patients. Proteinuria was significantly reduced from 151.7 +/- 162.4 mg/kg per 24 h at Week 0 to 36.9 +/- 42.3 at the end of the study in the group that received CSA (P < 0.05). There was no significant change in the proteinuria of the patients in the placebo group. A significant correlation between the percentage change of proteinuria over the 6 months of the study and the prestudy serum cholesterol levels (r = 0.79, P < 0.05) was seen in the CSA group. A partial correlation analysis controlling for the effects of serum cholesterol uncovered a significant relationship between average CSA level and proteinuria change (r = -0.76, P < 0.05). The fractional decline in GFR over the course of the study was not significantly different between the CSA and placebo-treated groups. In conclusion, CSA reduces proteinuria, increases serum albumin levels, and can be expected, therefore, to reduce the symptoms of nephrotic syndrome. Hypercholesterolemia antagonizes this effect of CSA.


Association between occupational exposure to mineral oil and rheumatoid arthritis: results from the Swedish EIRA case-control study.
The aim of the present study was to investigate the association between exposure to mineral oil and the risk of developing rheumatoid arthritis (RA), and in addition to perform a separate analysis on the major subphenotypes for the disease; namely, rheumatoid factor (RF)-positive RA, RF-negative RA, anticitrulline-positive RA and anticitrulline-negative RA, respectively. A population-based case-control study of incident cases of RA was performed among the population aged 18-70 years in a defined area of Sweden during May 1996-December 2003. A case was defined as an individual from the study base who for the first time received a diagnosis of RA according to the American College of Rheumatology criteria of 1987. Controls were randomly selected from the study base with consideration taken for age, gender and residential area. Cases (n = 1,419) and controls (n = 1,674) answered an extensive questionnaire regarding lifestyle factors and occupational exposures, including different types of mineral oils. Sera from cases and controls were investigated for RF and anticitrulline antibodies. Among men, exposure to any mineral oil was associated with a 30% increased relative risk of developing RA (relative risk = 1.3, 95% confidence interval = 1.0-1.7). When cases were subdivided into RF-positive RA and RF-negative RA, an increased risk was only observed for RF-positive RA (relative risk = 1.4, 95% confidence interval 1.0-2.0). When RA cases were subdivided according to the presence of anticitrulline antibodies, an increased risk associated with exposure to any mineral oil was observed only for anticitrulline-positive RA (relative risk = 1.6, 95% confidence interval = 1.1-2.2). Analysis of the interaction between oil exposure and the presence of HLA-DR shared epitope genes regarding the incidence of RA indicated that the increased risk associated with exposure to mineral oil was not related to the presence of shared epitope genotypes. In conclusion, our study shows that exposure to mineral oil is associated with an increased risk to develop RF-positive RA and anticitrulline-positive RA, respectively. The findings are of particular interest since the same mineral oils can induce polyarthritis in rats.


A Grounded Theory Qualitative Analysis of Interprofessional Providers' Perceptions on Caring for Critically Ill Infants and Children in Pediatric and General Emergency Departments.
The objective of this study was to explore pediatric emergency department (PED) and general emergency department (GED) providers' perceptions on caring for critically ill infants and children. This study utilized qualitative methods to examine the perceptions of emergency department providers caring for critically ill infants and children. Teams of providers participated in 4 in situ simulation cases followed by facilitated debriefings. Debriefings were recorded and professionally transcribed. The transcripts were reviewed independently and followed by group coding discussions to identify emerging themes. Consistent with grounded theory, the team iteratively revised the debriefing script as new understanding was gained. A total of 188 simulation debriefings were recorded in 24 departments, with 15 teams participating from 8 PEDs and 32 teams from 16 GEDs. Twenty-four debriefings were audiotaped and professionally transcribed verbatim. Thematic saturation was achieved after 20 transcripts. In our iterative qualitative analysis of these transcripts, we observed 4 themes: (1) GED provider comfort with algorithm-based pediatric care and overall comfort with pediatric care in PED, (2) GED provider reliance on cognitive aids versus experience-based recall by PED providers, (3) GED provider discomfort with locating and determining size or dose of pediatric-specific equipment and medications, and (4) PED provider reliance on larger team size and challenges with multitasking during resuscitation. Our qualitative analysis produced several themes that help us to understand providers' perceptions in caring for critically ill children in GEDs and PEDs. These data could guide the development of targeted educational and improvement interventions.


Targeting the epidermal growth factor receptor in non-small cell lung cancer cells: the effect of combining RNA interference with tyrosine kinase inhibitors or cetuximab.
The epidermal growth factor receptor (EGFR) is a validated therapeutic target in non-small cell lung cancer (NSCLC). However, current single agent receptor targeting does not achieve a maximal therapeutic effect, and some mutations confer resistance to current available agents. In the current study we have examined, in different NSCLC cell lines, the combined effect of RNA interference targeting the EGFR mRNA, and inactivation of EGFR signaling using different receptor tyrosine kinase inhibitors (TKIs) or a monoclonal antibody cetuximab. NSCLC cells (cell lines HCC827, H292, H358, H1650, and H1975) were transfected with EGFR siRNA and/or treated with the TKIs gefitinib, erlotinib, and afatinib, and/or with the monoclonal antibody cetuximab. The reduction of EGFR mRNA expression was measured by real-time quantitative RT-PCR. The down-regulation of EGFR protein expression was measured by western blot, and the proliferation, viability, caspase3/7 activity, and apoptotic morphology were monitored by spectrophotometry, fluorimetry, and fluorescence microscopy. The combined effect of EGFR siRNA and different drugs was evaluated using a combination index. EGFR-specific siRNA strongly inhibited EGFR protein expression almost equally in all cell lines and inhibited cell growth and induced cell apoptosis in all NSCLC cell lines studied, albeit with a different magnitude. The effects on growth obtained with siRNA was strikingly different from the effects obtained with TKIs. The effects of siRNA probably correlate with the overall oncogenic significance of the receptor, which is only partly inhibited by the TKIs. The cells which showed weak response to TKIs, such as the H1975 cell line containing the T790M resistance mutation, were found to be responsive to siRNA knockdown of EGFR, as were cell lines with downstream TKI resistance mutations. The cell line HCC827, harboring an exon 19 deletion mutation, was more than 10-fold more sensitive to TKI proliferation inhibition and apoptosis induction than any of the other cell lines. Cetuximab alone had no relevant in vitro activity at concentrations obtainable in the clinic. The addition of EGFR siRNA to either TKIs or cetuximab additively enhanced growth inhibition and induction of apoptosis in all five cell lines, independent of the EGFR mutation status (wild-type or sensitizing mutation or resistant mutation). The strongest biological effect was observed when afatinib was combined with an EGFR-specific siRNA. EGFR knockdown by siRNA further decreases the cell growth of lung cancer cells that are treated with TKIs or cetuximab alone, confirming that single agent drug targeting does not achieve a maximal biological effect. The siRNA inhibits EGFR oncogenic activity that bypasses downstream "resistance" mutations such as KRAS and PTEN. The combined treatment of siRNA and EGFR inhibitory agents is additive. The combination of a potent, irreversible kinase inhibitor such as afatinib, with EGFR-specific siRNAs should be further investigated as a new strategy in the treatment of lung cancer and other EGFR dependent cancers, including those with downstream resistance mutations.


Financial and quality impacts of the Medicare physician group practice demonstration.
To examine the impact of the Medicare Physician Group Practice (PGP) demonstration on expenditure, utilization, and quality outcomes. Secondary data analysis of 2001-2010 Medicare claims for 1,776,387 person years assigned to the ten participating provider organizations and 1,579,080 person years in the corresponding local comparison groups. We used a pre-post comparison group observational design consisting of four pre-demonstration years (1/01-12/04) and five demonstration years (4/05-3/10). We employed a propensity-weighted difference-in-differences regression model to estimate demonstration effects, adjusting for demographics, health status, geographic area, and secular trends. The ten demonstration sites combined saved $171 (2.0%) per assigned beneficiary person year (p<0.001) during the five-year demonstration period. Medicare paid performance bonuses to the participating PGPs that averaged $102 per person year. The net savings to the Medicare program were $69 (0.8%) per person year. Demonstration savings were achieved primarily from the inpatient setting. The demonstration improved quality of care as measured by six of seven claims-based process quality indicators. The PGP demonstration, which used a payment model similar to the Medicare Accountable Care Organization (ACO) program, resulted in small reductions in Medicare expenditures and inpatient utilization, and improvements in process quality indicators. Judging from this demonstration experience, it is unlikely that Medicare ACOs will initially achieve large savings. Nevertheless, ACOs paid through shared savings may be an important first step toward greater efficiency and quality in the Medicare fee-for-service program.


Unintegrated bovine leukemia virus DNA: association with viral expression and disease.
The correlation between bovine leukemia virus (BLV) unintegrated DNA, viral expression, and stage of disease was determined in cattle naturally infected with BLV. The concomitant presence of unintegrated BLV DNA with viral transcriptional activity was observed in 53% (18 of 34) of hematologically normal, BLV-seropositive cattle and in 100% (10 of 10) of BLV-seropositive cattle with the preneoplastic syndrome persistent lymphocytosis. In vitro studies suggested that accumulation of unintegrated BLV DNA resulted from a process of reinfection rather than intracellular reverse transcription of newly synthesized BLV RNA. Interestingly, unintegrated BLV DNA was not detected in tumor cells from cattle with BLV-associated lymphocytic leukemia/malignant lymphoma despite viral transcriptional activity in 100% (eight of eight) of these cattle. Thus, the presence of unintegrated BLV DNA differentiated nonneoplastic from neoplastic conditions in BLV-infected cattle. These results demonstrate that unintegrated viral DNA serves as a marker of disease progression in BLV-infected cattle but is not necessarily associated with induction or maintenance of the neoplastic state.


Purification of inkjet ink from water using liquid phase, electric discharge polymerization and cellulosic membrane filtration.
A method to separate inkjet ink from water was developed using a liquid phase, electric discharge process. The liquid phase, electric discharge process with filtration or sedimentation was shown to remove 97% of inkjet ink from solutions containing between 0.1-0.8 g/L and was consistent over a range of treatment conditions. Additionally, particle size analysis of treated allyl alcohol and treated propanol confirmed the electric discharge treatment has a polymerization mechanism, and small molecule analysis of treated methanol using gas chromatography and mass spectroscopy confirmed the mechanism was free radical initiated polymerization.


Effect of two forms of granulocyte-colony-stimulating factor on hepatic regeneration after 70% partial hepatectomy in rats.
1. The purpose of this study was to determine whether the commercially available forms of granulocyte-colony-stimulating factor exert the same beneficial effect on hepatic regeneration after 70% partial hepatectomy in rats. Adult male Wistar rats received either the two commercially available forms of granulocyte-colony-stimulating factor (Filgrastim or Lenograstim), or saline, simultaneously with partial hepatectomy. Hepatic regeneration was documented by determining [3H]thymidine incorporation into hepatic DNA, liver thymidine kinase activity, mitotic index and proliferating cell nuclear antigen immunostaining, at various time points after partial hepatectomy. 2. DNA biosynthesis, liver thymidine kinase activity and mitotic index of hepatocytes were not only enhanced (P < 0.001) in rats that received 150 micrograms of Filgrastim or Lenograstim/kg of body weight, but occurred earlier than in saline-treated partially hepatectomized rats. The administration of both forms of granulocyte-colony-stimulating factor, at the dose of 15 micrograms/kg of body weight, did not affect liver proliferative capacity, compared with observations in simply partially hepatectomized rats. High mitotic and proliferating cell nuclear antigen indices appeared earlier than those estimated in simply partially hepatectomized rats, when 150 micrograms of Filgrastim or Lenograstim/kg of body weight were administered. 3. These findings suggest that both pharmacologically available forms of granulocyte-colony-stimulating factor at a dose of 150 micrograms/kg of body weight are able to augment liver regenerative capacity, to the same extent, in this animal model of controlled hepatic proliferation.


Women's Experiences and Preferences in Relation to Infertility Counselling: A Multifaith Dialogue.
Religion and spirituality are a fundamental part of culture and influence how individuals experience and interpret infertility counselling. Thus far, little research has examined the influence of religiosity on the experience of infertility, and to our knowledge no study exists investigating the responses of religious infertile women to counselling. In this study we explored Muslim and Christian women's experiences and preferences with regard to infertility counselling. Using a grounded theory approach, 30 infertile women affiliated to different denominations of Islam (Shiite and Sunni) and Christianity (Protestantism, Catholicism, Orthodoxies) were interviewed. Data were collected through semi-structured in-depth interviews at fertility clinics in the UK and Iran, and analyzed using the Straussian mode of grounded theory. EMERGING CATEGORIES INCLUDED: Appraising the meaning of infertility religiously, applying religious coping strategies, and gaining a faith-based strength. These were encompassed in the core category of 'relying on a higher being'. Religious infertile women experienced infertility as an enriching experience for spiritual growth. This perspective helped them to acquire a feeling of self- confidence and strength to manage their emotions. Hence, they relied more on their own religious coping strategies and less on formal support resources like counselling services. However, they expected counsellors to be open to taking time to discuss their spiritual concerns in counselling sessions. In addition to focusing on clients' psychosocial needs, infertility counsellors should also consider religious and spiritual issues. Establishing a sympathetic and accepting relationship with infertile women will allow them to discuss their religious perspectives, which consequently may enhance their usage of counselling services.


Serum homocysteine, vitamin B12, and folate, and the prevalence and incidence of posterior subcapsular cataract.
We assessed associations between serum levels of homocysteine, vitamin B12, and folate, and the prevalence and 5-year incidence of posterior subcapsular cataract (PSC) in Blue Mountains Eye Study participants. We examined 3508 participants aged 49+ years during 1997 to 2000, including 2334 (75.1% of survivors) original and 1174 (85.2% of those eligible) newly recruited subjects. Five years later (2002-2004), 1952 (76.6% of survivors) original participants were re-examined. Detailed examinations, including lens photographs and fasting blood tests, were conducted at both visits. Logistic regression models estimated odds ratios (OR) and 95% confidence intervals (CI) after multivariable adjustment. In this population, those with PSC were older, less likely to have higher education, and more likely to have diabetes and myopia. The PSC prevalence was 5.7% (150/2644). Higher levels of homocysteine (per SD; OR, 1.17; 95% CI, 1.00-1.37) and lower levels of folate (per SD; OR, 1.24; 95% CI, 0.99-1.56) were associated with prevalent PSC. There was significant interaction (P < 0.05) between vitamin B12 and homocysteine; for B12 ≥125 pmol/L, 28% higher PSC prevalence was associated with homocysteine (per SD; OR, 1.28; 95% CI, 1.09-1.52); however, for B12 <125 pmol/L, nonsignificant lower PSC prevalence was associated with homocysteine (per SD; OR, 0.16; 95% CI, 0.02-1.57). The 5-year PSC incidence was 5.7% (n = 59/1030) with no significant associations with homocysteine, B12, and folate. Higher serum homocysteine level was associated with PSC prevalence in this population. Vitamin B12 status seemed to modify this association. Lack of longitudinal association could have resulted from insufficient study power.


Influence of Statistical Fluctuation on Reproducibility and Accuracy of SUVmax and SUVpeak: A Phantom Study.
Standardized uptake values (SUVs) have been widely used in the diagnosis of malignant tumors and in clinical trials of tumor therapies as semiquantitative metrics of tumor (18)F-FDG uptake. However, SUVs for small lesions are liable to errors due to partial-volume effect and statistical noise. The purpose of this study was to evaluate the reproducibility and accuracy of maximum and peak SUV (SUVmax and SUVpeak, respectively) of small lesions in phantom experiments. We used a body phantom with 6 spheres in a quarter warm background. The PET data were acquired for 1,800 s in list-mode, from which data were extracted to generate 15 PET images for each of the 60-, 90-, 120-, 150-, and 180-s scanning times. The SUVmax and SUVpeak of the hot spheres in the 1,800-s scan were used as a reference (SUVref,max and SUVref,peak). Coefficients of variation for both SUVmax and SUVpeak in hot spheres (CVmax and CVpeak) were calculated to evaluate the variability of the SUVs. On the other hand, percentage differences between SUVmax and SUVref,max and between SUVpeak and SUVref,peak were calculated for evaluation of the accuracy of SUV. We additionally examined the coefficients of variation of background activity and the percentage background variability as parameters for the physical assessment of image quality. Visibility of a 10-mm-diameter hot sphere was considerably different among scan frames. The CVmax and CVpeak increased as the sphere size became smaller and as the acquisition time became shorter. SUVmax was generally overestimated as the scan time shortened and the sphere size increased. The SUVmax and SUVpeak of a 37-mm-diameter sphere for 60-s scans had average positive biases of 28.3% and 4.4%, compared with the reference. SUVmax was variable and overestimated as the scan time decreased and the sphere size increased. In contrast, SUVpeak was a more robust and accurate metric than SUVmax. The measurements of SUVpeak (or SUVpeak normalized to lean body mass) in addition to SUVmax are desirable for reproducible and accurate quantification in clinical situations.


Lung EC-SOD overexpression attenuates hypoxic induction of Egr-1 and chronic hypoxic pulmonary vascular remodeling.
Although production of reactive oxygen species (ROS) such as superoxide (O(2)(.-)) has been implicated in chronic hypoxia-induced pulmonary hypertension (PH) and pulmonary vascular remodeling, the transcription factors and gene targets through which ROS exert their effects have not been completely identified. We used mice overexpressing the extracellular antioxidant enzyme extracellular superoxide dismutase (EC-SOD TG) to test the hypothesis that O(2)(.-) generated in the extracellular compartment under hypoxic conditions contributes to PH through the induction of the transcription factor, early growth response-1 (Egr-1), and its downstream gene target, tissue factor (TF). We found that chronic hypoxia decreased lung EC-SOD activity and protein expression in wild-type mice, but that EC-SOD activity remained five to seven times higher in EC-SOD TG mice under hypoxic conditions. EC-SOD overexpression attenuated chronic hypoxic PH, and vascular remodeling, measured by right ventricular systolic pressures, proliferation of cells in the vessel wall, muscularization of small pulmonary vessels, and collagen deposition. EC-SOD overexpression also prevented the early hypoxia-dependent upregulation of the redox-sensitive transcription factor Egr-1 and the procoagulant protein TF. These data provide the first evidence that EC-SOD activity is disrupted in chronic hypoxia, and increased EC-SOD activity can attenuate chronic hypoxic PH by limiting the hypoxic upregulation of redox-sensitive genes.


Development, validation and clinical impact of a prediction model for 6-month mortality in older cancer patients: the GRADE.
To develop, validate, and assess the clinical impact of a clinical score to predict a 6-month mortality risk among older cancer patients. The mean age was 81.2 ± 6.1 years (women: 54%, various cancers, metastatic cancer: 45%). The score, namely the GRADE, included two geriatric variables (unintentional weight loss, impaired mobility), two oncological variables (cancer site, cancer extension), and exclusively supportive care. Up to a 14% risk of early death, the decision curves suggest that cancer treatment should be instated. We have developed and validated a simple score, easy to implement in daily oncological practice, to predict early death among older cancer patients which could guide oncologists in their treatment decisions. 603 outpatients prospectively included in the Physical Frailty in Elderly Cancer patients cohort study. We created a multivariate prediction model by evaluating the strength of the individual components of the Geriatric Assessment regarding risk of death at 6 months. Each component was evaluated by univariate analysis and the significant variables (P ≤ 0.20) were carried on as covariates in the multivariate cox proportion hazard analysis. The beta coefficients from the model were used to build a point-based scoring system. Clinical impact was assessed using decision curves.


Downstaging to non-invasive urothelial carcinoma is associated with improved outcome following radical cystectomy for patients with cT2 disease.
Pathologic stage is a critically important prognostic factor after radical cystectomy (RC) that is used to guide the use of secondary therapies. However, the risk of disease recurrence, for patients clinically diagnosed with muscle-invasive tumors who are found not to have muscle-invasive disease at RC are poorly defined. Therefore, we reviewed the long-term outcomes in patients who were downstaged to non-invasive urothelial carcinoma at time of RC. We identified 1,177 consecutive patients with muscle-invasive urothelial carcinoma of the bladder who underwent radical cystectomy at our institution between 1980 and 1999 without neoadjuvant therapy. Postoperative disease recurrence and survival were estimated using the Kaplan-Meier method and compared using the log rank test. Cox proportional hazard regression models were used to analyze the impact of pathologic stage on survival. Pathologic downstaging to non-muscle invasive disease was identified in 538 (45.7 %) patients. The 10-year cancer-specific survival was 84.1, 77.4, 71.1 and 58.5 % for those with pT0, pTis, pT1 and pT2 tumors, respectively. On multivariate analysis, the risk of cancer-specific mortality was significantly decreased for patients with non-muscle invasive disease than those with organ-confined muscle invasion (RR-0.39; p = 0.002). There was no difference in disease-specific mortality among patients who had non-invasive (pT0, pTa, or pTis) disease (p = 0.19). Downstaging from clinical muscle-invasive bladder cancer to non-muscle invasive disease at RC is associated with a significant reduction in cancer-specific mortality. However, even patients with residual non-muscle invasive disease may suffer disease recurrence and require continued surveillance after surgery.


Selection with partial selfing. I. Mass selection.
The expected responses to mass selection carried out before or after reproduction in a population whose members all have a fixed probability of self pollination (s) are formulated using covariances of relatives and their component quadratic functions for a model with arbitrary additive and dominance effects. The response measured in the first generation offspring after selection (immediate gain) can differ from that retained when the population has regained equilibrium (permanent gain). The population mean behaves in a predictable manner during the return to equilibrium, and its value at any time can be predicted from earlier generations. The permanent gain from selection after reproduction is always (1 + s)/2 times as large as that from selection before reproduction, but the relationship of the immediate gains depends on the genetic model assumed. Numerical analysis applied to a model with two alleles per locus and varying allele frequencies, dominance ratios and numbers of loci showed that the proportion of the immediate gain retained at equilibrium was reduced with the large inbreeding depression associated with increasing dominance levels and numbers of loci and was generally lower for selection after reproduction than before. In the absence of information as to the magnitude of genetic variances and inbreeding depression in species reproducing by partial selfing, the importance of this phenomenon is unknown.


Molecular characterization of Chlamydomonas reinhardtii telomeres and telomerase mutants.
Telomeres are repeated sequences found at the end of the linear chromosomes of most eukaryotes and are required for chromosome integrity. Expression of the reverse-transcriptase telomerase allows for extension of telomeric repeats to counteract natural telomere shortening. Although Chlamydomonas reinhardtii, a photosynthetic unicellular green alga, is widely used as a model organism in photosynthesis and flagella research, and for biotechnological applications, the biology of its telomeres has not been investigated in depth. Here, we show that the C. reinhardtii (TTTTAGGG)n telomeric repeats are mostly nondegenerate and that the telomeres form a protective structure, with a subset ending with a 3' overhang and another subset presenting a blunt end. Although telomere size and length distributions are stable under various standard growth conditions, they vary substantially between 12 genetically close reference strains. Finally, we identify CrTERT, the gene encoding the catalytic subunit of telomerase and show that telomeres shorten progressively in mutants of this gene. Telomerase mutants eventually enter replicative senescence, demonstrating that telomerase is required for long-term maintenance of telomeres in C. reinhardtii.


Socioeconomic inequalities in homicide mortality: a population-based comparative study of 12 European countries.
Recent research has suggested that violent mortality may be socially patterned and a potentially important source of health inequalities within and between countries. Against this background the current study assessed socioeconomic inequalities in homicide mortality across Europe. To do this, longitudinal and cross-sectional data were obtained from mortality registers and population censuses in 12 European countries. Educational level was used to indicate socioeconomic position. Age-standardized mortality rates were calculated for post, upper and lower secondary or less educational groups. The magnitude of inequalities was assessed using the relative and slope index of inequality. The analysis focused on the 35-64 age group. Educational inequalities in homicide mortality were present in all countries. Absolute inequalities in homicide mortality were larger in the eastern part of Europe and in Finland, consistent with their higher overall homicide rates. They contributed 2.5% at most (in Estonia) to the inequalities in total mortality. Relative inequalities were high in the northern and eastern part of Europe, but were low in Belgium, Switzerland and Slovenia. Patterns were less consistent among women. Socioeconomic inequalities in homicide are thus a universal phenomenon in Europe. Wide-ranging social and inter-sectoral health policies are now needed to address the risk of violent victimization that target both potential offenders and victims.


Eye- and neck/shoulder-discomfort during visually demanding experimental near work.
Frequent use of digital information technology has an impact on eye- and neck/shoulder-discomfort. Studies with cross sectional and intervention design indicate an association between the two symptom categories. Still, whether visually demanding near work, per se, contributes to increased neck/shoulder discomfort remains a question of debate. The aim of this laboratory study was to assess if visually demanding experimental near work affects eye- and neck/shoulder-discomfort when the posture was adjusted for comfort and no movements were allowed. Thirty-three healthy subjects performed a visually demanding computer screen task (viewing task) under four different optical lens conditions: binocular -3.5 D and monocular -3.5 D, +3.5 D and ± 0 D. During the experiment subjects were seated in an office chair (with neck support) that was individually adjusted for comfort. At baseline and after each viewing task, subjects reported their perceived eye- and neck/shoulder discomfort on Borg's CR-10 scale. Results show a significant increase of eye discomfort between baseline and the first viewing task, and a significant increase in neck/shoulder discomfort from baseline throughout the first three viewing tasks. Further analysis is required to determent whether the neck/shoulder discomfort was induced by the demanding near work or the static posture, or a combination.


Relationship between caesium (137Cs) load, cardiovascular symptoms, and source of food in 'Chernobyl' children -- preliminary observations after intake of oral apple pectin.
Seventeen years after the nuclear power accident at Chernobyl, most of the radio-contamination among the population of Southern Belarus is caused by incorporation of long-lived radioisotopes. The varying levels of 137Cs observed among children in this area are explained by the source of their food, especially by the consumption of contaminated milk produced privately. We stratified children from rural areas of Belarus (caesium [137Cs] contamination >5 Ci/km(2)) by their 137Cs loads into three distinct groups (group 1, <5 Bq/kg body weight [BW]; group 2, 38.4 +/- 2.4 Bq/kg BW; group 3, 122 +/- 18.5 Bq/kg BW). We determined the relationship between the 137Cs load and the children's main source of food and recorded their cardiovascular symptoms. Cardiovascular symptoms, ECG alterations, and arterial hypertension were significantly more frequent in children with high 137Cs burden than in children with very low 137Cs burden. Children with moderate and high 137Cs loads (groups 2 and 3) received apple pectin, a food additive, for 16 days. Apple pectin significantly decreased 137Cs loads in these groups (39% and 28%, respectively). ECG alterations improved, while cardiovascular symptoms and hypertension did not change in any group.


Epidemiology of febrile diseases in the emergency department of a Caribbean island: the Curaçao experience.
The aetiology of febrile diseases in tropical countries often remains poorly characterized. We aim to describe the aetiology and outcome of febrile illnesses at the Emergency Department (ED) in Curaçao. From April 2008 - April 2009, all adult febrile patients (T> 38.5 degrees C) at the ED of the St Elisabeth Hospital, Curaçao, Netherlands Antilles, were included. Clinical data were recorded, routine laboratory measurements and blood cultures were taken. Final diagnoses were made at discharge by an independent physician and in retrospect by the main investigator Four hundred and three patients were included: 223 patients (55.6%) were hospitalized, 32 patients (7.9%) died and 18 patients (4.5%) were admitted to the Intensive Care Unit. In 129 febrile patients (32.0%), infection was proven; 84.4% of patients had bacterial (29.0% urinary tract infection, 23.2% pneumonia infection), 5.6% viral and 10.0% parasitic or fungal infections. Twenty-one patients (5.2%) were discharged with a non-infectious diagnosis and 172 patients (42.7%) without a clear diagnosis. A high mortality rate of 7.9% was observed. We found a high prevalence of bacterial infections, with pneumonia and urinary tract infections as the most common causes of fever. One in 20 patients did not have an infectious disease.


Diagnostic Efficacy of Molecular Techniques for Detection and Identification of Leishmania Species in Human Whole Blood and Skin Samples from Ecuador.
Microscopic examination is the standard method for diagnosis of cutaneous and mucocutaneous leishmaniasis despite its low sensitivity. This study compared the diagnosis efficacy of microscopic examination versus polymerase chain reaction (PCR)-based methods and DNA sequencing using whole blood and skin lesion samples from patients with suspected leishmaniasis. The presence of Leishmania was determined by microscopy and amplification of 18S ribosomal RNA gene from blood and skin samples of 22 patients. Twenty individuals were positive for leishmaniasis. Microscopic analysis identified 85%, whereas PCR identified 100% of positive cases from skin and 90% from blood. Cytochrome b gene (cyt-b) amplification and sequencing identified Leishmania guyanensis, Leishmania shawi, and Leishmania naiffi from skin and blood samples. This study demonstrated the usefulness of whole blood and molecular techniques for the diagnosis and species identification of leishmaniasis.


Identification and utilization of arbitrary correlations in models of recombination signal sequences.
A significant challenge in bioinformatics is to develop methods for detecting and modeling patterns in variable DNA sequence sites, such as protein-binding sites in regulatory DNA. Current approaches sometimes perform poorly when positions in the site do not independently affect protein binding. We developed a statistical technique for modeling the correlation structure in variable DNA sequence sites. The method places no restrictions on the number of correlated positions or on their spatial relationship within the site. No prior empirical evidence for the correlation structure is necessary. We applied our method to the recombination signal sequences (RSS) that direct assembly of B-cell and T-cell antigen-receptor genes via V(D)J recombination. The technique is based on model selection by cross-validation and produces models that allow computation of an information score for any signal-length sequence. We also modeled RSS using order zero and order one Markov chains. The scores from all models are highly correlated with measured recombination efficiencies, but the models arising from our technique are better than the Markov models at discriminating RSS from non-RSS. Our model-development procedure produces models that estimate well the recombinogenic potential of RSS and are better at RSS recognition than the order zero and order one Markov models. Our models are, therefore, valuable for studying the regulation of both physiologic and aberrant V(D)J recombination. The approach could be equally powerful for the study of promoter and enhancer elements, splice sites, and other DNA regulatory sites that are highly variable at the level of individual nucleotide positions.


Specific binding of perhydrohistrionicotoxin to Torpedo acetylcholine receptor.
Torpedo californica postsynaptic membrane fragments were treated with base, which resulted in membranes that were depleted of many nonacetylcholine receptor polypeptides and contained acetylcholine receptor subunits of Mr 40,000, 50,000, 60,000, and 65,000 (Raftery, M.A., Vandlen, R.L., Reed, K.L. & Lee T. (1975) Cold Spring Harbor Symp. Quant. Biol. 40, 193-202). A 43,000-Mr polypeptide and some other components were quantitatively extracted. Base-treated membranes retained the capacity to bind [3H]perhydrohistrionicotoxin and the local anesthetics dibucaine and tetracaine. The regulation of this binding by carbamylcholine, as well as the kinetic mechanism of perhydrohistrionicotoxin binding, was unchanged. [3H]Perhydrohistrionicotoxin binding activity was largely reconstituted from 2% sodium cholate extracts of base-treated membranes. Therefore, the perhydrostrionicotoxin binding site appears to be located on one or more of the acetylcholine receptor polypeptides, and the reconstitution of that binding site from detergent extracts does not require the presence of a 43,000-Mr polypeptide.


Antifouling activity of portimine, select semisynthetic analogues, and other microalga-derived spirocyclic imines.
A range of natural products from marine invertebrates, bacteria and fungi have been assessed as leads for nature-inspired antifouling (AF) biocides, but little attention has been paid to microalgal-derived compounds. This study assessed the AF activity of the spirocyclic imine portimine (1), which is produced by the benthic mat-forming dinoflagellate Vulcanodinium rugosum. Portimine displayed potent AF activity in a panel of four macrofouling bioassays (EC50 0.06-62.5 ng ml-1), and this activity was distinct from that of the related compounds gymnodimine-A (2), 13-desmethyl spirolide C (3), and pinnatoxin-F (4). The proposed mechanism of action for portimine is induction of apoptosis, based on the observation that portimine inhibited macrofouling organisms at developmental stages known to involve apoptotic processes. Semisynthetic modification of select portions of the portimine molecule was subsequently undertaken. Observed changes in bioactivity of the resulting semisynthetic analogues of portimine were consistent with portimine's unprecedented 5-membered imine ring structure playing a central role in its AF activity.


Immunoglobulin E-binding autoantigens: biochemical characterization and clinical relevance.
Although immediate-Type I skin reactions to human dander have been described six decades ago, only the recent application of molecular biology to allergology research allowed fast and detailed characterization of IgE-binding autoantigens. These can be functionally subdivided into three classes: (1) self-antigens with sequence homology to environmental allergens belonging to the class of phylogenetically conserved proteins, (2) self-antigens without sequence homology to known environmental allergens, and (3) chemically modified self-antigens deriving from workplace exposure. As environmental allergens, also IgE-binding autoantigens belong to different protein families without common structural features that would explain their IgE-binding capability. Many of the self-antigens showing sequence homology to environmental allergens, are phylogenetically conserved proteins like manganese dependent superoxide dismutase, thioredoxin or cyclopilin. Their IgE-binding capability can be explained by molecular mimicry resulting from shared B-cell epitopes. A common factor of IgE-binding self-antigens without sequence homology to known environmental allergens is that they elicit IgE responses only in individuals suffering from long-lasting atopic diseases. In contrast, IgE-mediated reactions to modified self-antigens might be explained with the generation of novel B-cell epitopes. Chemically modified self-antigens are likely to be recognized as non-self by the immune system. The clinical relevance of IgE responses to self-antigens remains largely unclear. Well documented is their ability to induce immediate Type I skin reactions in vivo, and to induce mediator release from effector cells of sensitized individuals in vitro. Based on these observations it is reasonable to assume that IgE-mediated cross-linking of FcRIε receptors on effector cells can elicit the same symptoms as those induced by environmental allergens, and this could explain exacerbations of chronic allergic diseases in the absence of external exposure. However, because most of the described IgE-binding self-antigens are intracellular proteins normally not accessible for antigen-antibody interactions, local release of the antigens is required to explain the induction of symptoms.


A computerized decision support system for depression in primary care.
In 2004, results from The Texas Medication Algorithm Project (TMAP) showed better clinical outcomes for patients whose physicians adhered to a paper-and-pencil algorithm compared to patients who received standard clinical treatment for major depressive disorder (MDD). However, implementation of and fidelity to the treatment algorithm among various providers was observed to be inadequate. A computerized decision support system (CDSS) for the implementation of the TMAP algorithm for depression has since been developed to improve fidelity and adherence to the algorithm. This was a 2-group, parallel design, clinical trial (one patient group receiving MDD treatment from physicians using the CDSS and the other patient group receiving usual care) conducted at 2 separate primary care clinics in Texas from March 2005 through June 2006. Fifty-five patients with MDD (DSM-IV criteria) with no significant difference in disease characteristics were enrolled, 32 of whom were treated by physicians using CDSS and 23 were treated by physicians using usual care. The study's objective was to evaluate the feasibility and efficacy of implementing a CDSS to assist physicians acutely treating patients with MDD compared to usual care in primary care. Primary efficacy outcomes for depression symptom severity were based on the 17-item Hamilton Depression Rating Scale (HDRS(17)) evaluated by an independent rater. Patients treated by physicians employing CDSS had significantly greater symptom reduction, based on the HDRS(17), than patients treated with usual care (P < .001). The CDSS algorithm, utilizing measurement-based care, was superior to usual care for patients with MDD in primary care settings. Larger randomized controlled trials are needed to confirm these findings. clinicaltrials.gov Identifier: NCT00551083.


Impact of γ-rays Irradiation on Hybrid TiO2-SiO2 Sol-Gel Films Doped with RHODAMINE 6G.
In the present paper, we investigate how the optical and structural properties, in particular the observed photoluminescence (PL) of photocurable and organic-inorganic TiO2-SiO2 sol-gel films doped with Rhodamine 6G (R6G) are affected by γ-rays. For this, four luminescent films, firstly polymerized with UV photons (365 nm), were submitted to different accumulated doses of 50 kGy, 200 kGy, 500 kGy and 1 MGy while one sample was kept as a reference and unirradiated. The PL, recorded under excitations at 365 nm, 442 nm and 488 nm clearly evidences that a strong signal peaking at 564 nm is still largely present in the γ-irradiated samples. In addition, M-lines and Fourier-transform infrared (FTIR) spectroscopies are used to quantify the radiation induced refractive index variation and the chemical changes, respectively. Results show that a refractive index decrease of 7 × 10-3 at 633 nm is achieved at a 1 MGy accumulated dose while a photo-induced polymerization occurs, related to the consumption of CH=C, Si-OH and Si-O-CH3 groups to form Ti-O and Si-O bonds. All these results confirm that the host matrix (TiO2-SiO2) and R6G fluorophores successfully withstand the hard γ-ray exposure, opening the way to the use of this material for sensing applications in radiation-rich environments.


Genome-wide association meta-analysis of functional outcome after ischemic stroke.
To discover common genetic variants associated with poststroke outcomes using a genome-wide association (GWA) study. The study comprised 6,165 patients with ischemic stroke from 12 studies in Europe, the United States, and Australia included in the GISCOME (Genetics of Ischaemic Stroke Functional Outcome) network. The primary outcome was modified Rankin Scale score after 60 to 190 days, evaluated as 2 dichotomous variables (0-2 vs 3-6 and 0-1 vs 2-6) and subsequently as an ordinal variable. GWA analyses were performed in each study independently and results were meta-analyzed. Analyses were adjusted for age, sex, stroke severity (baseline NIH Stroke Scale score), and ancestry. The significance level was p < 5 × 10-8. We identified one genetic variant associated with functional outcome with genome-wide significance (modified Rankin Scale scores 0-2 vs 3-6, p = 5.3 × 10-9). This intronic variant (rs1842681) in the LOC105372028 gene is a previously reported trans-expression quantitative trait locus for PPP1R21, which encodes a regulatory subunit of protein phosphatase 1. This ubiquitous phosphatase is implicated in brain functions such as brain plasticity. Several variants detected in this study demonstrated suggestive association with outcome (p < 10-5), some of which are within or near genes with experimental evidence of influence on ischemic stroke volume and/or brain recovery (e.g., NTN4, TEK, and PTCH1). In this large GWA study on functional outcome after ischemic stroke, we report one significant variant and several variants with suggestive association to outcome 3 months after stroke onset with plausible mechanistic links to poststroke recovery. Future replication studies and exploration of potential functional mechanisms for identified genetic variants are warranted.


Hepatocyte transplantation--in vitro cytotoxic reaction of autologous granulocytes and mononuclears to isolated hepatocytes.
Hepatocyte (HC) transplantation (tx) may be useful for bridging patients to whole organ transplantation and for providing metabolic support during liver failure and for replacing whole organ transplantation in certain liver metabolic diseases. In specific situations where the death rate of host hepatocytes is high, the transplanted cells can repopulate the native liver. Successful transplantation of hepatocytes is hampered by lack of proper cellular (stromal) and humoral (cytokines) environment at the site of implantation. We have found that another factor responsible for low in vivo survival rate of transplanted HC is their rapid destroying by host granulocytes and monocytes. AIM. In this study we investigated the in vitro process of destruction of HC by granulocytes and mononuclear cells, the phenotypes of effector mononuclears and the tempo of HC lysis. In vitro cell-mediated cytotoxicity, HC-PMN and HC-PBM rosette formation rate and HC lysis, as well as phenotypes of HC-adhering cells were investigated. Granulocytes formed rosettes with HC almost immediately after the beginning of incubation and were found highly cytotoxic to HC. The cellular mechanism of lysis was not mediated by serum natural antibodies. Also the in vitro mixed HC-granulocyte 51Cr test showed high granulocyte cytolytic activity. Monoclonal antibodies to class I and II antigens, CD11/18 and 54 did not block the granulocyte cytotoxicity. Blood mononuclear cells also formed rosettes with HC and were cytotoxic to them, but the level of cytotoxicity was lower than of granulocytes. ED1+ monocytes revealed highest cytolytic activity toward HC. Hepatocyte contained only trace levels of endotoxin and no chemotactic activity of granulocytes and monocytes toward HC could be observed. A random physical contact of blood leukocytes seems necessary for adhesion to isolated HC. Taken together, granulocytes and monocytes recognize intercellular surface molecules on HC "exposed during isolation" from the hepatic trabeculae as "non-self" and lyse HC by direct contact.


Effects of varying approaches for identifying respiratory disturbances on sleep apnea assessment.
Varying approaches to measuring the respiratory disturbance index (RDI) may lead to discrepant estimates of the severity of sleep-disordered breathing (SDB). In this study, we assessed the impact of varying the use of corroborative data (presence and degree of desaturation and/or arousal) to identify hypopneas and apneas. The relationships among 10 RDIs defined by various definitions of apneas and hypopneas were assessed in 5,046 participants in the Sleep Heart Health Study (SHHS) who underwent overnight unattended 12-channel polysomnography (PSG). The magnitude of the median RDI varied 10-fold (i.e., 29.3 when the RDI was based on events identified on the basis of flow or volume amplitude criteria alone to 2.0 for an RDI that required an associated 5% desaturation with events). The correlation between RDIs based on different definitions ranged from 0.99 to 0.68. The highest correlations were among RDIs that required apneas and hypopneas to be associated with some level of desaturation. Lower correlations were observed between RDIs that required desaturation as compared with RDIs defined on the basis of amplitude criteria alone or associated arousal. These data suggest that different approaches for measuring the RDI may contribute to substantial variability in identification and classification of the disorder.


Effect of basic fibroblast growth factor on angiogenesis and growth of isografted bone: quantitative in vitro-in vivo analysis in mice.
Basic fibroblast growth factor (bFGF), a constituent of bone and cartilage matrix, has been shown to be a potent mitogen for osteoblasts and chondrocytes and yet an inhibitor of chondrocyte terminal differentiation in cell culture. To characterize the effect of bFGF on bone formation, whole neonatal murine femora were cultured in the presence or absence of bFGF and a neutralizing antibody against bFGF. In vitro, femoral elongation was provided by cartilage growth only; the calcified diaphyseal zone stained by oxytetracycline did not increase. When bFGF was added to the culture medium, longitudinal growth of the proximal and distal cartilage was inhibited in a dose-dependent manner (p < 0.05), and the number of hypertrophic chondrocytes in the growth plate was reduced. This phenomenon was absent in the presence of a neutralizing antibody, which when given alone significantly promoted femoral elongation. In contrast, in vivo after transplantation into adult mice bearing dorsal skin fold chambers, femora rapidly calcified after revascularization. This observation supports the notion that bone formation largely depends on angiogenesis-mediated events. To verify this hypothesis, angiogenesis and bone formation were quantified using bFGF known to be a stimulator of angiogenesis. Calcification of grafted femora was accelerated by bFGF given intraperitoneally. The neutralizing antibody slightly suppressed angiogenesis and femoral elongation (not statistically significant), whereas intravenous injections of both substances did not reveal a significant modulatory effect. In vivo the effect of systemically administered bFGF was inhomogeneous, but there was a strong correlation between angiogenesis and endochondral calcification (p < 0.001). These results suggest that exogenous bFGF modulates bone formation in vitro by inhibition of terminal differentiation of chondrocytes in the growth plate, and angiogenesis and concomitant in vivo events are pivotal in the promotion of rapid bone formation.


Bilateral Internal Thoracic Artery Composite Y Grafts: Analysis of 464 Angiograms in 296 Patients.
The use of bilateral internal thoracic arteries (BITA) in coronary artery bypass graft surgery improves patient survival. The optimum BITA graft configuration is unknown. This study evaluates outcomes after the use of bilateral internal mammary Y grafting as the primary grafting strategy. Patients who underwent BITA composite Y grafting (n = 1,011) between October 1994 and March 2009 were identified from a cardiac surgical database. Follow-up (mean 12 years) was obtained by cross-reference with the state death registry and local cardiology databases. Perioperative mortality was 1.6%. Kaplan-Meier 10-year survival was 87.2%. There were 464 episodes of postdischarge angiography in 296 patients, at an average of 5.5 years to first angiography. Graft failure occurred in 151 patients, predominately affecting the right ITA limb (n = 139). Degree of native vessel stenosis (odds ratio [OR] 0.66 per 10% increase), anatomic territory grafted (circumflex OR 2.64, right coronary OR 6.73 versus anterior), and end-to-side free wall anastomoses (OR 1.98) predicted anastomotic occlusion. The left anterior descending artery graft patency was unaffected by sequential grafting. Progression to graft failure after the first angiogram was rarely seen in the 111 patients who had serial angiography. Outcomes of BITA Y grafting are similar to those for other BITA configurations. The rate of presentation with arterial graft dysfunction decreases over time. Although competitive flow affects anastomotic patency, there is no threshold at which risk of occlusion substantially increases.


In vitro gap-directed translesion DNA synthesis of an abasic site involving human DNA polymerases epsilon, lambda, and beta.
DNA polymerase (pol) ε is thought to be the leading strand replicase in eukaryotes, whereas pols λ and β are believed to be mainly involved in re-synthesis steps of DNA repair. DNA elongation by the human pol ε is halted by an abasic site (apurinic/apyrimidinic (AP) site). In this study, we present in vitro evidence that human pols λ, β, and η can perform translesion synthesis (TLS) of an AP site in the presence of pol ε, likely by initiating the 3'OHs created at the lesion by the arrested pol ε. However, in the case of pols λ and β, this TLS requires the presence of a DNA gap downstream from the product synthesized by the pol ε, and the optimal gap for efficient TLS is different for the two polymerases. The presence of gaps did not affect the TLS capacity of human pol η. Characterization of the reaction products showed that pol β inserted dAMP opposite the AP site, whereas gap filling synthesis by pol λ resulted in single or double deletions opposite the lesion. The synthesis up to the AP site by pol ε and the subsequent TLS by pols λ and β are not influenced by human processivity factor proliferating cell nuclear antigen and human single-stranded DNA-binding protein replication protein A. The bypass capacity of pol λ at the AP site is greatly reduced when a truncated form of the enzyme, which has lost the BRCA1 C-terminal and proline-rich domains, is used. Collectively, our in vitro results support the existence of a mechanism of gap-directed TLS at an AP site involving a switch between the replicative pol ε and the repair pols λ and β.


Access to specialized pediatric cancer care in Switzerland.
Specialized pediatric cancer centers (PCCs) are thought to be essential to obtain state-of-the-art care for children and adolescents. We determined the proportion of childhood cancer patients not treated in a PCC, and described their characteristics and place of treatment. The Swiss Childhood Cancer Registry (SCCR) registers all children treated in Swiss PCCs. The regional cancer registries (covering 14/26 cantons) register all cancer patients of a region. The children of the SCCR with data from 7 regions (11 cantons) were compared, using specialized software for record linkage. All children <16 years of age at diagnosis with primary malignant tumors, diagnosed between 1990 and 2004, and living in one of these regions were included in the analysis. 22.1% (238/1,077) of patients recorded in regional registries were not registered in the SCCR. Of these, 15.7% (169/1,077) had never been in a PCC while 6.4% (69/1,077) had been in a PCC but were not registered in the SCCR, due to incomplete data flow. In all diagnostic groups and in all age groups, a certain proportion of children was treated outside a PCC, but this proportion was largest in children suffering from malignant bone tumors/soft tissue sarcomas and from malignant epithelial neoplasms, and in older children. The proportion of patients treated in a PCC increased over the study period (P < 0.0001). One in six childhood cancer patients in Switzerland was not treated in a PCC. Whether these patients have different treatment outcomes remained unclear.


Olive oil intake and risk of cardiovascular disease and mortality in the PREDIMED Study.
It is unknown whether individuals at high cardiovascular risk sustain a benefit in cardiovascular disease from increased olive oil consumption. The aim was to assess the association between total olive oil intake, its varieties (extra virgin and common olive oil) and the risk of cardiovascular disease and mortality in a Mediterranean population at high cardiovascular risk. We included 7,216 men and women at high cardiovascular risk, aged 55 to 80 years, from the PREvención con DIeta MEDiterránea (PREDIMED) study, a multicenter, randomized, controlled, clinical trial. Participants were randomized to one of three interventions: Mediterranean Diets supplemented with nuts or extra-virgin olive oil, or a control low-fat diet. The present analysis was conducted as an observational prospective cohort study. The median follow-up was 4.8 years. Cardiovascular disease (stroke, myocardial infarction and cardiovascular death) and mortality were ascertained by medical records and National Death Index. Olive oil consumption was evaluated with validated food frequency questionnaires. Multivariate Cox proportional hazards and generalized estimating equations were used to assess the association between baseline and yearly repeated measurements of olive oil intake, cardiovascular disease and mortality. During follow-up, 277 cardiovascular events and 323 deaths occurred. Participants in the highest energy-adjusted tertile of baseline total olive oil and extra-virgin olive oil consumption had 35% (HR: 0.65; 95% CI: 0.47 to 0.89) and 39% (HR: 0.61; 95% CI: 0.44 to 0.85) cardiovascular disease risk reduction, respectively, compared to the reference. Higher baseline total olive oil consumption was associated with 48% (HR: 0.52; 95% CI: 0.29 to 0.93) reduced risk of cardiovascular mortality. For each 10 g/d increase in extra-virgin olive oil consumption, cardiovascular disease and mortality risk decreased by 10% and 7%, respectively. No significant associations were found for cancer and all-cause mortality. The associations between cardiovascular events and extra virgin olive oil intake were significant in the Mediterranean diet intervention groups and not in the control group. Olive oil consumption, specifically the extra-virgin variety, is associated with reduced risks of cardiovascular disease and mortality in individuals at high cardiovascular risk. This study was registered at controlled-trials.com (http://www.controlled-trials.com/ISRCTN35739639). International Standard Randomized Controlled Trial Number (ISRCTN): 35739639. Registration date: 5 October 2005.


Autocatalytic RNA cleavage in the human beta-globin pre-mRNA promotes transcription termination.
New evidence indicates that termination of transcription is an important regulatory step, closely related to transcriptional interference and even transcriptional initiation. However, how this occurs is poorly understood. Recently, in vivo analysis of transcriptional termination for the human beta-globin gene revealed a new phenomenon--co-transcriptional cleavage (CoTC). This primary cleavage event within beta-globin pre-messenger RNA, downstream of the poly(A) site, is critical for efficient transcriptional termination by RNA polymerase II. Here we show that the CoTC process in the human beta-globin gene involves an RNA self-cleaving activity. We characterize the autocatalytic core of the CoTC ribozyme and show its functional role in efficient termination in vivo. The identified core CoTC is highly conserved in the 3' flanking regions of other primate beta-globin genes. Functionally, it resembles the 3' processive, self-cleaving ribozymes described for the protein-encoding genes from the myxomycetes Didymium iridis and Physarum polycephalum, indicating evolutionary conservation of this molecular process. We predict that regulated autocatalytic cleavage elements within pre-mRNAs may be a general phenomenon and that functionally it may provide the entry point for exonucleases involved in mRNA maturation, turnover and, in particular, transcriptional termination.


Osteosarcoma arising in fibrous dysplasia, confirmed by mutational analysis of GNAS gene.
Malignancy arising in fibrous dysplasia (FD) is rare. Approximately 100 cases have been reported so far, and osteosarcoma is the most common malignancy. We report a case of osteosarcoma in a 33-year-old Japanese man with monostotic FD of the right proximal femur from the age of 16 years. Histologically, relatively well-differentiated osteosarcoma was found in the FD lesion. Immunohistochemically, the FD was negative for p53 or MDM2, and the MIB-1 index was less than 1%, whereas the osteosarcoma was positive for both p53 and MDM2, and the MIB-1 index was up to 15%. The FD and osteosarcoma were negative for CDK4. Fluorescent in situ hybridization assay showed no amplification of the MDM2 gene, indicating that the osteosarcoma was a conventional osteosarcoma, not an intraosseous well-differentiated type. The original cell of malignancy in FD is unclear. Malignancy can be potentially derived from dysplastic cells in the area of the FD or cells in the adjacent normal tissues. GNAS gene mutation has recently been reported for fibrous dysplasia and the mutation is highly specific to fibrous dysplasia among fibro-osseous lesions including osteosarcoma. In this case, point mutations of GNAS were found in the FD and osteosarcoma but not in the adjacent normal tissues, suggesting that osteosarcoma was derived from the spindle cells of FD. This is the first report to clearly show that osteosarcoma is derived from the spindle cells in fibrous dysplasia (FD).


Association between beta-blocker use and fracture risk: the Dubbo Osteoporosis Epidemiology Study.
In animal model, mice treated with beta-blockers (BB) had increased bone mass. In humans, high bone mass is associated with reduce fracture risk. The present study sought to test the hypothesis that BB use is associated with reduced fracture risk. Data from 3488 participants (1285 men) aged 50 years and above in the Dubbo Osteoporosis Epidemiology Study (DOES) were analyzed. Baseline characteristics of participants were obtained at the initial visit which had taken place between 1989 and 1993. Bone mineral density (BMD) at the lumbar spine and femoral neck was measured by dual energy X-ray absorptiometry (GE-LUNAR Corp, Madison, WI). Two hundred and sixty two (20%) men and 411 (19%) women had been on BB, as ascertained by direct interview and verification with medication history. The incidence of fragility fractures was ascertained during the follow-up period (1989-2008). In men, BB use was associated with higher BMD at the femoral neck (0.96 versus 0.92 g/cm², P < 0.01), higher lumbar spine (1.32 versus 1.25 g/cm², P < 0.01), and lower fracture risk than those not on BB (odds ratio [OR]: 0.49; 95% CI: 0.32-0.75). In women, BB users also had higher femoral neck BMD (0.83 versus 0.81 g/cm², P < 0.01), higher lumbar spine BMD (1.11 versus 1.06 g/cm², P < 0.01), and lower risk of fracture than non-users (OR 0.68, 95% CI: 0.53-0.87). The associations between BB use and fracture risk were independent of age, BMD, and clinical risk factors. Subgroup analysis suggested that the association was mainly found in selective BB, not in non-selective BB. Beta-blockers use, particularly selective BB, was associated with reduced fracture risk in both men and women, and the association was independent of BMD.


A Single Nucleotide Deletion in Gibberellin20-oxidase1 Causes Alpine Dwarfism in Arabidopsis.
Alpine dwarfism is widely observed in alpine plant populations and often considered a high-altitude adaptation, yet its molecular basis and ecological relevance remain unclear. In this study, we used map-based cloning and field transplant experiments to investigate dwarfism in natural Arabidopsis (Arabidopsis thaliana) accessions collected from the Swiss Alps. A loss-of-function mutation due to a single nucleotide deletion in gibberellin20-oxidase1 (GA5) was identified as the cause of dwarfism in an alpine accession. The mutated allele, ga5-184, was found in two natural Arabidopsis populations collected from one geographic region at high altitude, but was different from all other reported ga5 null alleles, suggesting that this allele has evolved locally. In field transplant experiments, the dwarf accession with ga5-184 exhibited a fitness pattern consistent with adaptation to high altitude. Across a wider array of accessions from the Swiss Alps, plant height decreased with altitude of origin, but fitness patterns in the transplant experiments were variable and general altitudinal adaptation was not evident. In general, our study provides new insights into molecular basis and possible ecological roles of alpine dwarfism, and demonstrates the importance of the GA-signaling pathway for the generation of ecologically relevant variation in higher plants.