The CATIE and CUtLASS studies in schizophrenia: results and implications for clinicians.
Numerous double-blind studies have compared second-generation antipsychotics (SGAs) with first-generation antipsychotics (FGAs), with most finding better efficacy and tolerability for SGAs. However, these 'efficacy trials' were generally short term and included only highly selected patients. Mostly because of weight gain and other metabolic effects of the SGAs, as well as their high acquisition price, the debate on the (cost) effectiveness of the SGAs led to two pragmatic clinical trials with no sponsorship by industry. Both trials had broad inclusion criteria and long follow-up, and tried to mimic clinical routine: CATIE (Clinical Antipsychotic Trials of Intervention Effectiveness) and CUtLASS (Cost Utility of the Latest Antipsychotic drugs in Schizophrenia Study). 1493 patients participated in CATIE, an 18-month, double-blind trial comparing the SGAs olanzapine, quetiapine, risperidone and ziprasidone with the FGA perphenazine. If efficacy or tolerability was insufficient, patients were re-randomized to a medication other than the one they previously received. Improvement of psychopathology and of quality of life was only moderate. Overall, 74% of patients discontinued study medication before 18 months, and the median time to discontinuation was 4.6 months. Aside from olanzapine (time to discontinuation 9.2 months), the other SGAs did not differ from each other or from perphenazine. Except for adverse effects as a reason for discontinuation, differences between the SGAs and the FGA were minimal. In CUtLASS, a 12-month open-label trial, 277 patients were randomized to receive an FGA or a SGA. Again, efficacy was rather similar between the two groups, with only limited improvement of psychopathology and quality of life. The authors of both trials concluded that SGAs do not markedly differ from FGAs regarding compliance, quality of life and effectiveness. The methodological problems of both trials have been discussed extensively. Patients had psychotic symptoms that were moderate in severity and were at least partially treatment resistant. The marginal improvement observed indicated that this population might not be appropriate to detect differences between FGAs and SGAs. Specific issues of CATIE include the exclusion of patients with tardive dyskinesia in the perphenazine arm and the high discontinuation rate. In CUtLASS, the concept of including 13 different FGAs and four SGAs in the respective classes was problematic. It is of interest that the most widely prescribed drug was sulpiride--of the FGAs, this is probably the 'most atypical' drug. Aside from the finding that the advantages of the SGAs are not as strong as early trials and marketing suggested or promised, the trials do not provide much helpful information regarding everyday practice. For tardive dyskinesia, no conclusions at all can be drawn. Similarly, methodological problems inhibited the detection of the other major advantage of the SGAs, i.e. the improved subjective well-being/quality of life while receiving these agents. It is well known that patients' and doctors' perspectives differ markedly, and the Quality of Life Scale (QLS), an expert-rated scale used in both trials, might not be sensitive enough to detect the subjective advantages reported by the majority of patients in other trials. CATIE and CUtLASS suggest that SGAs do not live up to all the previous expectations. However, even if most of these advantages are debatable, the lower risk of tardive dyskinesia and the better subjective effects should be strong enough reasons to favour these drugs. There is no single antipsychotic that is best for every schizophrenia patient, as individual responses differ markedly. For successfully individualized treatment, a multitude of antipsychotic options are needed.


Latent Profile Analysis of the Five Facet Mindfulness Questionnaire in a Sample With a History of Recurrent Depression.
Extending previous research, we applied latent profile analysis in a sample of adults with a history of recurrent depression to identify subgroups with distinct response profiles on the Five Facet Mindfulness Questionnaire and understand how these relate to psychological functioning. The sample was randomly divided into two subsamples to first examine the optimal number of latent profiles (test sample; n = 343) and then validate the identified solution (validation sample; n = 340). In both test and validation samples, a four-profile solution was revealed where two profiles mapped broadly onto those previously identified in nonclinical samples: "high mindfulness" and "nonjudgmentally aware." Two additional subgroups, "moderate mindfulness" and "very low mindfulness," were observed. "High mindfulness" was associated with the most adaptive psychological functioning and "very low mindfulness" with the least adaptive. In most people with recurrent depression, mindfulness skills are expressed evenly across different domains. However, in a small minority a meaningful and replicable uneven profile indicating nonjudgmental awareness is observable. Current findings require replication and future research should examine the extent to which profiles change from periods of wellness to illness in people with recurrent depression and how profiles are influenced by exposure to mindfulness-based intervention.


Evolution of Protein Quaternary Structure in Response to Selective Pressure for Increased Thermostability.
Oligomerization has been suggested to be an important mechanism for increasing or maintaining the thermostability of proteins. Although it is evident that protein-protein contacts can result in substantial stabilization in many extant proteins, evidence for evolutionary selection for oligomerization is largely indirect and little is understood of the early steps in the evolution of oligomers. A laboratory-directed evolution experiment that selected for increased thermostability in the αE7 carboxylesterase from the Australian sheep blowfly, Lucilia cuprina, resulted in a thermostable variant, LcαE7-4a, that displayed increased levels of dimeric and tetrameric quaternary structure. A trade-off between activity and thermostability was made during the evolution of thermostability, with the higher-order oligomeric species displaying the greatest thermostability and lowest catalytic activity. Analysis of monomeric and dimeric LcαE7-4a crystal structures revealed that only one of the oligomerization-inducing mutations was located at a potential protein-protein interface. This work demonstrates that by imposing a selective pressure demanding greater thermostability, mutations can lead to increased oligomerization and stabilization, providing support for the hypothesis that oligomerization is a viable evolutionary strategy for protein stabilization.


Ascending retrocecal appendicitis: clinical and computed tomographic findings.
To describe the clinical presentation and computed tomographic (CT) features of ascending retrocecal appendicitis. During the past 8 years, 1670 patients with surgically proven appendicitis were identified by a retrospective investigation. Thirty-three patients who underwent contrast-enhanced abdominal CT before surgery and had ascending retrocecal appendicitis at surgery were included in the patient cohort. The clinical and CT findings of these 33 patients were analyzed. Patients presented with right lower abdominal pain (49%, 16/33), right flank pain (24%, 8/33), right upper abdominal pain (18%, 6/33), and periumbilical pain (15%, 5/33). Inflamed ascending retrocecal appendices were visualized completely in 70% (23/33), partially in 21% (7 of 33), and not detected in the remaining 9% (3/33). Periappendiceal inflammatory changes were most commonly observed in the retrocolic space in 88% (29/33), followed by paracolic gutter (30%, 10/33), pararenal space (27%, 9/33), mesentery (24%, 8/33), perirenal space (18%, 6/33), and subhepatic space (3%, 1/33). Inflammatory thickening of Gerota fascia (70%, 23/33) and the lateroconal fascia (64%, 21/33) was observed as well. Perforation of the appendix with the formation of an abscess was present in 49% (16/33). The abscesses were most commonly located in the retrocolic space (88%, 14/16). Diffuse wall thickenings of the cecum (67%, 22/33) and the ascending colon (64%, 21/33) were often observed also. Appendicoliths were found in 33% (11/33). More than half of patients with ascending retrocecal appendicitis presented with atypical clinical presentation. At CT, ascending retrocecal appendicitis was associated with a high incidence of retroperitoneal inflammatory changes and appendiceal perforation.


Analysis of the gene expression profile activated by the CC chemokine ligand 5/RANTES and by lipopolysaccharide in human monocytes.
The gene expression profile induced by the CC chemokine ligand (CCL) 5/RANTES in human monocytes was examined using the oligonucleotide array technology. Of 5600 transcripts examined, 42 were consistently induced by CCL5, and none were suppressed. Chemokine-inducible transcripts could be clustered in functional groups, including selected cytokines and receptors (e.g., IL-1beta, CCL2/monocyte chemotactic protein-1, and the CCL5 receptor CCR1) and molecules involved in extracellular matrix recognition and digestion (e.g., CD44 splice transcripts, urokinase-type plasminogen activator receptor, matrix metalloprotease (MMP)-9, and MMP-19). Transcript expression, confirmed by quantitative real-time PCR analysis for selected genes, was associated with protein induction for some (e.g., CCL2), but not all (e.g., IL-1beta), transcripts examined. The chemokine-induced gene profile was distinct from that activated by LPS, a prototypic phagocyte activator. Although certain transcripts were stimulated by both agonists (e.g., IL-1beta and CCL2), others were induced only by either LPS (e.g., TNF-alpha and IL-6) or CCL5 (e.g., MMP-19) or were divergently regulated (e.g., CCR1). Thus, CCL5, a prototypic CC inflammatory chemokine, activates a restricted transcriptional program in monocytes distinct from that induced by the prototypic pathogen-derived proinflammatory stimulant LPS. Chemokine-induced chemokines production could represent a novel amplification loop of leukocyte recruitment, while a subset of chemokine-inducible transcripts could be involved in monocyte extravasation and tissue invasion.


Nuclear localization of the interferon-inducible protein kinase PKR in human cells and transfected mouse cells.
The levels and subcellular distribution of the interferon-inducible double-stranded RNA-dependent protein kinase PKR have been measured in human Daudi cells and stably transfected mouse NIH 3T3 cells expressing the human protein kinase. Immunofluorescence of intact cells and quantitative immunoblotting of cell extracts indicate that PKR occurs in both the cytoplasm and the cell nucleus, with staining specifically in the nucleolus. The ratio of cytoplasmic to nuclear PKR is approximately 5:1 in control cells; in response to interferon treatment the protein kinase is induced severalfold in the cytoplasm whereas the level in the nucleus does not increase significantly. Analysis of individual transfected cells by confocal microscopy reveals a pattern of distribution of PKR similar to that in Daudi cells, with immunostaining of cytoplasm and nucleoli. Similar results are observed whether cells expressing wild-type PKR or a catalytically inactive mutant form of the kinase are analyzed, but untransfected 3T3 cells are not stained by the antibody used. Two-dimensional isoelectric focusing analysis of PKR in whole cell extracts reveals the presence of multiple forms with different pI values whereas similar analysis of the nuclear fraction indicates only one predominant species with a relatively basic pI. These results suggest that PKR may have a role in the cell nucleus as well as the cytoplasm and that the subcellular distribution of the protein kinase may be related to post-translational modifications.


Social-ecological landscape patterns predict woody encroachment from native tree plantings in a temperate grassland.
Afforestation is often viewed as the purposeful planting of trees in historically nonforested grasslands, but an unintended consequence is woody encroachment, which should be considered part of the afforestation process. In North America's temperate grassland biome, Eastern redcedar (Juniperus virginiana L.) is a native species used in tree plantings that aggressively invades in the absence of controlling processes. Cedar is a well-studied woody encroacher, but little is known about the degree to which cedar windbreaks, which are advocated for in agroforestry programs, are contributing to woody encroachment, what factors are associated with cedar spread from windbreaks, nor where encroachment from windbreaks is occurring in contemporary social-ecological landscapes. We used remotely sensed imagery to identify the presence and pattern of woody encroachment from windbreaks in the Nebraska Sandhills. We used multimodel inference to compare three classes of models representing three hypotheses about factors that could influence cedar spread: (a) windbreak models based on windbreak structure and design elements; (b) abiotic models focused on local environmental conditions; and (c) landscape models characterizing coupled human-natural features within the broader matrix. Woody encroachment was evident for 23% of sampled windbreaks in the Nebraska Sandhills. Of our candidate models, our inclusive landscape model carried 92% of the model weight. This model indicated that encroachment from windbreaks was more likely near roadways and less likely near farmsteads, other cedar plantings, and waterbodies, highlighting strong social ties to the distribution of woody encroachment from tree plantings across contemporary landscapes. Our model findings indicate where additional investments into cedar control can be prioritized to prevent cedar spread from windbreaks. This approach can serve as a model in other temperate regions to identify where woody encroachment resulting from temperate agroforestry programs is emerging.


Cancer vaccines using supramolecular hydrogels of NSAID-modified peptides as adjuvants abolish tumorigenesis.
Inflammatory responses play crucial roles in the development and progression of tumors. Tumor-associated inflammation not only promotes tumor growth but also induces the suppression of immune responses against tumors. We demonstrate in this study that hydrogels of nonsteroidal anti-inflammatory drug (NSAID) modified D-tetrapeptides (GDFDFDY) are promising cancer vaccine adjuvants, especially for Fbp-gel and Car-gel. The hydrogels allow easy incorporation of a protein OVA antigen by vortexing. Our results indicate that vaccines based on Fbp-gel and Car-gel increase IgG production by 1476- and 929-fold, compared with the OVA group, respectively. They exhibit higher IgG2a antibody titers and stimulate the production of IFN-γ and IL-6 cytokines. Their higher antibody and cytokine eliciting properties in combination with their anti-inflammatory properties endow them with excellent tumor elimination properties in vivo. In a preventive immune assay against B16-OVA tumors, they totally prevent tumorigenesis. In a therapeutic immune assay against EG7-OVA tumors, they inhibit tumor growth by 75%, compared with the PBS group. Our results suggest the great potential of our hydrogels in the development of vaccines to treat cancers.


Structure and stability of symptoms in first episode psychosis: a longitudinal network approach.
Early psychosis is characterised by heterogeneity in illness trajectories, where outcomes remain poor for many. Understanding psychosis symptoms and their relation to illness outcomes, from a novel network perspective, may help to delineate psychopathology within early psychosis and identify pivotal targets for intervention. Using network modelling in first episode psychosis (FEP), this study aimed to identify: (a) key central and bridge symptoms most influential in symptom networks, and (b) examine the structure and stability of the networks at baseline and 12-month follow-up. Data on 1027 participants with FEP were taken from the National EDEN longitudinal study and used to create regularised partial correlation networks using the 'EBICglasso' algorithm for positive, negative, and depressive symptoms at baseline and at 12-months. Centrality and bridge estimations were computed using a permutation-based network comparison test. Depression featured as a central symptom in both the baseline and 12-month networks. Conceptual disorganisation, stereotyped thinking, along with hallucinations and suspiciousness featured as key bridge symptoms across the networks. The network comparison test revealed that the strength and bridge centralities did not differ significantly between the two networks (C = 0.096153; p = 0.22297). However, the network structure and connectedness differed significantly from baseline to follow-up (M = 0.16405, p = <0.0001; S = 0.74536, p = 0.02), with several associations between psychosis and depressive items differing significantly by 12 months. Depressive symptoms, in addition to symptoms of thought disturbance (e.g. conceptual disorganisation and stereotyped thinking), may be examples of important, under-recognized treatment targets in early psychosis, which may have the potential to lead to global symptom improvements and better recovery.


Supplementation of hydrogen-rich water improves lipid and glucose metabolism in patients with type 2 diabetes or impaired glucose tolerance.
Oxidative stress is recognized widely as being associated with various disorders including diabetes, hypertension, and atherosclerosis. It is well established that hydrogen has a reducing action. We therefore investigated the effects of hydrogen-rich water intake on lipid and glucose metabolism in patients with either type 2 diabetes mellitus (T2DM) or impaired glucose tolerance (IGT). We performed a randomized, double-blind, placebo-controlled, crossover study in 30 patients with T2DM controlled by diet and exercise therapy and 6 patients with IGT. The patients consumed either 900 mL/d of hydrogen-rich pure water or 900 mL of placebo pure water for 8 weeks, with a 12-week washout period. Several biomarkers of oxidative stress, insulin resistance, and glucose metabolism, assessed by an oral glucose tolerance test, were evaluated at baseline and at 8 weeks. Intake of hydrogen-rich water was associated with significant decreases in the levels of modified low-density lipoprotein (LDL) cholesterol (ie, modifications that increase the net negative charge of LDL), small dense LDL, and urinary 8-isoprostanes by 15.5% (P < .01), 5.7% (P < .05), and 6.6% (P < .05), respectively. Hydrogen-rich water intake was also associated with a trend of decreased serum concentrations of oxidized LDL and free fatty acids, and increased plasma levels of adiponectin and extracellular-superoxide dismutase. In 4 of 6 patients with IGT, intake of hydrogen-rich water normalized the oral glucose tolerance test. In conclusion, these results suggest that supplementation with hydrogen-rich water may have a beneficial role in prevention of T2DM and insulin resistance.


The role of curvature anisotropy in the ordering of spheres on an ellipsoid.
Non-spherical emulsion droplets can be stabilized by densely packed colloidal particles adsorbed at their surface. In order to understand the microstructure of these surface packings, the ordering of hard spheres on ellipsoidal surfaces is determined through large scale computer simulations. Defects in the packing are shown generically to occur most often in regions of strong curvature; however, the relationship between defects and curvature is nontrivial, and the distribution of defects shows secondary maxima for ellipsoids of sufficiently high aspect ratio. As with packings on spherical surfaces, additional defects beyond those required by topology are observed as chains or "scars". The transition point, however, is found to be softened by the anisotropic curvature which also partially orients the scars. A rich library of symmetric commensurate packings are identified for low particle number. We verify experimentally that ellipsoidal droplets of varying aspect ratio can be arrested by surface-adsorbed colloids.


The Xenopus homolog of Drosophila Suppressor of Hairless mediates Notch signaling during primary neurogenesis.
The X-Notch-1 receptor, and its putative ligand, X-Delta-1, are thought to mediate an inhibitory cell-cell interaction, called lateral inhibition, that limits the number of primary neurons that form in Xenopus embryos. The expression of Xenopus ESR-1, a gene related to Drosophila Enhancer of split, appears to be induced by Notch signaling during this process. To determine how the activation of X-Notch-1 induces ESR-1 expression and regulates primary neurogenesis, we isolated the Xenopus homolog of Suppressor of Hairless (X-Su(H)), a component of the Notch signaling pathway in Drosophila. Using animal cap assays, we show that X-Su(H) induces ESR-1 expression, perhaps directly, when modified by the addition of ankyrin repeats. Using a DNA binding mutant of X-Su(H), we show that X-Su(H) activity is required for induction of ESR-1. Finally, expression of the DNA binding mutant in embryos leads to a neurogenic phenotype as well as increased expression of both X-Delta-1 and XNGNR1, a proneural gene expressed during primary neurogenesis. These results suggest that activation of X-Su(H) is a key step in the Notch signaling pathway during primary neurogenesis in Xenopus embryos.


Equity in the allocation of public sector financial resources in low- and middle-income countries: a systematic literature review.
This review aims to identify, assess and analyse the evidence on equity in the distribution of public health sector expenditure in low- and middle-income countries. Four bibliographic databases and five websites were searched to identify quantitative studies examining equity in the distribution of public health funding in individual countries or groups of countries. Two different types of studies were identified: benefit incidence analysis (BIA) and resource allocation comparison (RAC) studies. Quality appraisal and data synthesis were tailored to each study type to reflect differences in the methods used and in the information provided. We identified 39 studies focusing on African, Asian and Latin American countries. Of these, 31 were BIA studies that described the distribution, typically across socio-economic status, of individual monetary benefit derived from service utilization. The remaining eight were RAC studies that compared the actual expenditure across geographic areas to an ideal need-based distribution. Overall, the quality of the evidence from both types of study was relatively weak. Looking across studies, the evidence confirms that resource allocation formulae can enhance equity in resource allocation across geographic areas and that the poor benefits proportionally more from primary health care than from hospital expenditure. The lack of information on the distribution of benefit from utilization in RAC studies and on the countries' approaches to resource allocation in BIA studies prevents further policy analysis. Additional research that relates the type of resource allocation mechanism to service provision and to the benefit distribution is required for a better understanding of equity-enhancing resource allocation policies.


Physicians' knowledge and attitudes regarding periconceptional folic acid supplementation: a survey in Southern Israel.
Pre-conceptional folic acid supplementation is an effective way to reduce the incidence of neural tube defects (NTDs). Primary care providers are an important source of information to promote folic acid intake. This study aimed at evaluating primary care physicians' knowledge and attitudes regarding folic acid supplementation for childbearing women. A questionnaire on physicians' knowledge and attitudes, mostly including multiple-choice questions, was delivered by mail to all physicians (n=370) in a large health provider organization in southern Israel in 2006. Data regarding demographics as well as type of specialty, experience, and place of work were collected. Eighty-seven physicians were included in the study (response rate: 24%). Seventy physicians out of 81 (94%) reported routine recommendation of folic acid for their patients. Most physicians admitted that they needed more information regarding folic acid supplementation. Knowledge about folic acid's role in preventing congenital anomalies was suboptimal, with 2% of the physicians correctly estimating the efficacy of folic acid in decreasing the risk of NTDs and 8% recognizing the association between folic acid supplementation and decreased prevalence of malformations other than NTDs. Knowledge about the correct timing (12%) and dosage (47%) of folic acid preparations for average-risk women was also lacking. Primary care physicians report that they routinely prescribe folic acid to women in childbearing age in order to prevent congenital anomalies, but their knowledge about folic acid supplementation is insufficient.


Phyloclimatic modeling: combining phylogenetics and bioclimatic modeling.
We investigate the impact of past climates on plant diversification by tracking the "footprint" of climate change on a phylogenetic tree. Diversity within the cosmopolitan carnivorous plant genus Drosera (Droseraceae) is focused within Mediterranean climate regions. We explore whether this diversity is temporally linked to Mediterranean-type climatic shifts of the mid-Miocene and whether climate preferences are conservative over phylogenetic timescales. Phyloclimatic modeling combines environmental niche (bioclimatic) modeling with phylogenetics in order to study evolutionary patterns in relation to climate change. We present the largest and most complete such example to date using Drosera. The bioclimatic models of extant species demonstrate clear phylogenetic patterns; this is particularly evident for the tuberous sundews from southwestern Australia (subgenus Ergaleium). We employ a method for establishing confidence intervals of node ages on a phylogeny using replicates from a Bayesian phylogenetic analysis. This chronogram shows that many clades, including subgenus Ergaleium and section Bryastrum, diversified during the establishment of the Mediterranean-type climate. Ancestral reconstructions of bioclimatic models demonstrate a pattern of preference for this climate type within these groups. Ancestral bioclimatic models are projected into palaeo-climate reconstructions for the time periods indicated by the chronogram. We present two such examples that each generate plausible estimates of ancestral lineage distribution, which are similar to their current distributions. This is the first study to attempt bioclimatic projections on evolutionary time scales. The sundews appear to have diversified in response to local climate development. Some groups are specialized for Mediterranean climates, others show wide-ranging generalism. This demonstrates that Phyloclimatic modeling could be repeated for other plant groups and is fundamental to the understanding of evolutionary responses to climate change.


The Healthy Eating Index 2005 and risk for pancreatic cancer in the NIH-AARP study.
Dietary pattern analyses characterizing combinations of food intakes offer conceptual and statistical advantages over food- and nutrient-based analyses of disease risk. However, few studies have examined dietary patterns and pancreatic cancer risk and none focused on the 2005 Dietary Guidelines for Americans. We used the Healthy Eating Index 2005 (HEI-2005) to estimate the association between meeting those dietary guidelines and pancreatic cancer risk. We calculated the HEI-2005 score for 537 218 men and women in the National Institutes of Health-American Association of Retired Persons Diet and Health Study using responses to food frequency questionnaires returned in 1995 and 1996. We used Cox proportional hazards regression to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for risk of pancreatic cancer according to HEI-2005 quintiles and explored effect modification by known risk factors. P interaction values were calculated using the Wald test. All statistical tests were two-sided. We identified 2383 incident, exocrine pancreatic cancer cases (median = 10.5 years follow-up). Comparing participants who met the most dietary guidelines (Q5) with those who met the fewest guidelines (Q1), we observed a reduced risk of pancreatic cancer (HR = 0.85, 95% CI = 0.74 to 0.97). Among men there was an interaction by body mass index (P interaction = .03), with a hazard ratio of 0.72 (95% CI = 0.59 to 0.88) comparing Q5 vs Q1 in overweight/obese men (body mass index ≥ 25 kg/m(2)) but no association among normal weight men. Our findings support the hypothesis that consuming a high-quality diet, as scored by the HEI-2005, may reduce the risk of pancreatic cancer.


Premature thelarche: identification of clinical and laboratory data for the diagnosis of precocious puberty.
Two groups of girls with premature breast development were studied retrospectively. We tried to identify clinical, radiological, and hormonal parameters that could distinguish between a benign, nonprogressive premature thelarche and a true precocious puberty. The clinical outcome of 88 girls with breast enlargement before 6.1 years of age was analyzed. Taking into account the progression of their sexual maturation, we allocated the children into 2 groups: "Isolated Premature Thelarche" (n = 63) and "Precocious Puberty" (n = 25) groups. Chronological and bone ages, height and growth velocity centiles, computerized tomography of hypothalamus-pituitary area, pelvic ultrasonography, gonadotropin response to luteinizing hormone-releasing hormone stimulation as well as basal levels of luteinizing hormone, follicle-stimulating hormone, estradiol, and prolactin were studied in both groups. Statistical analysis were performed using the Student t test to compare the sample means. Fisher's exact test and chi2 test were used to analyze the nonparametric variables. Isolated premature thelarche most frequently affected girls younger than 2 years who presented exaggerated follicle-stimulating hormone response to luteinizing hormone-releasing hormone stimulation test. The precocious puberty group had higher initial stature, accelerated growth rate and bone age, increased uterine and ovarian volumes, high spontaneous luteinizing hormone levels by immunofluorimetric assay, as well as a high luteinizing hormone response and peak luteinizing hormone/follicle-stimulating hormone ratio after luteinizing hormone-releasing hormone stimulation. At initial presentation, girls who undergo true precocious puberty present advanced bone age, increased uterine and ovarian volumes in addition to breast enlargement, as well as an luteinizing hormone-predominant response after a luteinizing hormone-releasing hormone stimulation test.


Surveys on therapeutic effects of "halotherapy chamber with artificial salt-mine environment" on patients with certain chronic allergenic respiratory pathologies and infectious-inflammatory pathologies.
Halotherapy (HT), derived from speleotherapy in salt mines, is also a drug-free therapeutic method. HT effects vary depending on the therapeutic method and the structure of halotherapy environment. The purpose of this article is to show the HT effects of "halotherapy chamber with artificial salt-mine environment" of the National Institute of Rehabilitation, Physical Medicine and Balneoclimatology (INRMFB), on patients with bronchial asthma and other chronic, infectious-inflammatory and allergic respiratory diseases, describing the clinical effects on certain nonspecific resistance factors, on markers of inflammatory processes and on certain immunological changes. Patients were clinically assessed, with the application of hematologic investigations, analysis of nonspecific resistance to infection and of inflammatory process markers, immunologic assessments, analysis of sodium and potassium concentrations, of mineralocorticoid function and other biochemical tests. For the experimental HT therapy performed in the "halotherapy chamber with artificial salt-mine environment" of INRMFB, 15 patients suffering from bronchial asthma, allergic rhinitis, chronic bronchitis, chronic obstructive bronchopneumopathy were selected, based on specific medical indications and contraindications and applying ethical principles, as well as 4 patients with similar pathologies for the control group, who underwent in-home drug treatment. After the specific halotherapy treatment on patients with bronchial asthma, chronic bronchitis and chronic obstructive bronchopneumopathy, which also showed other chronic, infectious-inflammatory and allergic respiratory pathologies, triggering of anti-inflammatory (and also anti allergic) mechanisms and healing effects on inflammatory process were noted. Data acquired also proved the halo therapeutic effect causing the reduction of sensitiveness of body in patients with bronchial asthma.


Management to reduce nitrogen losses in animal production.
Reduction of nitrogen loss in animal production requires whole-farm management. Reduced loss from one farm component is easily negated in another if all components are not equally well managed. Animal excretion of manure N can be decreased by improving the balance of protein or amino acids fed to that required by individual animals or animal groups or by improving production efficiency. Management to increase milk, meat, or egg production normally improves efficiency by reducing the maintenance protein required per unit of production. Large losses of manure nitrogen occur through the ammonia and nitrous oxide that are emitted into the atmosphere and the nitrate leached into groundwater. Up to half of the excreted nitrogen is lost from the housing facility, but this loss can be decreased through frequent manure removal and by avoiding deep litter systems and feedlots. Techniques such as acid treatment of manure, scrubbing of ventilation air, and floor designs for separating feces and urine substantially reduce ammonia emissions, but these practices are often impractical or uneconomical for general use. Manure storage units improve nutrient utilization by allowing better timing of nutrient application with crop needs. At least 70% of the nitrogen entering anaerobic lagoons is typically lost, but a less than 10% loss can be maintained using slurry storage with a natural crust or other cover, or by drying poultry manure to at least 50% dry matter. Irrigation and surface spreading of manure without soil incorporation often ensures the loss of all remaining nonorganic nitrogen (typically, 20 to 40% of remaining nitrogen). Rapid incorporation and shallow injection methods decrease this loss by at least 50%, and deep injection into the soil essentially eliminates this loss. For grazing animals, excessive loss can be avoided by not overstocking pastures and avoiding late fall and winter grazing. Reducing emissions between the animal and the soil can lead to greater leaching and denitrification losses from the soil if this additional nitrogen is not used properly. The use of a crop rotation that efficiently absorbs these nutrients and applying nitrogen near the time it is needed by crops reduce the potential for further loss. Maintaining the proper number of animals per unit of land available for manure application is always crucial for efficient recycling of nitrogen. Our understanding of nitrogen loss processes is improved through modeling, and computer models assist in the development of integrated systems for efficient and economical nitrogen use in animal production.


Polyclonality of familial murine adenomas: analyses of mouse chimeras with low tumor multiplicity suggest short-range interactions.
In previous studies demonstrating the polyclonal structure of familial intestinal adenomas, high tumor multiplicity made it difficult to eliminate the possibility that polyclonality arose by the random collision of distinct initiated clones as opposed to some form of clonal interaction. We sought to test further the random collision hypothesis. Chimeric mice carrying the multiple intestinal neoplasia (Min) mutation of the adenomatous polyposis coli gene (Apc) and homozygous for the tumor resistance allele of the Mom1 locus were established. These chimeras also display a strong propensity for tumors of polyclonal structure, despite their markedly reduced tumor multiplicity. Considering tumor sizes and multiplicities, the observed fraction of overtly polyclonal heterotypic adenomas was significantly higher than predicted by the random collision hypothesis. This finding supports models of polyclonality involving interaction among multiple initiated clones. The extent of clonal interaction was assessed by statistical analyses that relate the observed frequency of overtly polyclonal heterotypic tumors to the geometry of the chimeric patches and the pattern of underlying crypts. These statistical calculations indicate that the familial adenomas of the Apc(Min/+) mouse may commonly form through interactions between clones as close as 1-2 crypt diameters apart.


The development of nutritional-supplement fact sheets for Irish athletes: a case study.
The use of sport supplements presents a dilemma for many of those involved in supporting athletes, including coaches, families, support staff, and the athletes themselves. Often the information that they source can be incorrect and promote a biased view regarding the use of nutritional supplements. The aim of this case study was to describe the process that occurred around the development of a series of targeted educational fact sheets on a range of nutritional supplements for Irish athletes. It describes the initiation and support of the process by the Irish Sports Council; one of its subgroups, the Food and Food Supplements Committee; and the Irish Institute of Sport. A needs assessment through questionnaires was carried out to establish the most commonly used sport nutrition supplements by athletes age 16 or over in Ireland. Respondents completed 105 questionnaires over a 4-mo period in 2008-09 that led to the production of 20 supplement fact sheets. These supplement fact sheets will enable Irish athletes to access high-quality, up-to-date, scientific information about the supplements they have reported consuming. Since personal reading had a strong influence over athletes' decision-making process for taking nutritional supplements, as did scientific research, fact sheets available on the Internet from a reliable source are an ideal way to educate Irish athletes.


ROC methodology within a monitoring framework.
Receiver operating characteristic (ROC) methodology is widely used to evaluate and compare diagnostic tests. Generally, each diagnostic test is applied once to each subject in a population and the results, reported on a continuous scale, are used to construct the ROC curve. We extend the standard method to accommodate a framework in which the diagnostic test is repeated over time to monitor for occurrence of an event. Unlike the usual situation in which event status is static, the problem we address involves event status that is not constant over the monitoring period. Subjects generally are classified as non-events, or controls, until they experience events that convert them to cases. Viewing the data as incomplete discrete failure time data with time-varying covariates, potentially useful diagnostic markers can be related appropriately in time with the true condition and varying amounts of information per individual can be taken into account. The ROC curve provides an assessment of the performance of the test in combination with the schedule of testing. Within this framework, a computational simplification is introduced to calculate variances and covariances for the areas under the ROC curves. Periodic monitoring for reperfusion following thrombolytic treatment for acute myocardial infarction provides a detailed example, whereby the lengths of the testing interval combined with different diagnostic markers are compared.


Improving Adherence to Secondary Stroke Prevention Strategies Through Motivational Interviewing: Randomized Controlled Trial.
Stroke recurrence rates are high (20%-25%) and have not declined over past 3 decades. This study tested effectiveness of motivational interviewing (MI) for reducing stroke recurrence, measured by improving adherence to recommended medication and lifestyle changes compared with usual care. Single-blind, prospective phase III randomized controlled trial of 386 people with stroke assigned to either MI treatment (4 sessions at 28 days, 3, 6, and 9 months post stroke) or usual care; with outcomes assessed at 28 days, 3, 6, 9, and 12 months post stroke. Primary outcomes were change in systolic blood pressure and low-density lipoprotein cholesterol levels as indicators of adherence at 12 months. Secondary outcomes included self-reported adherence, new stroke, or coronary heart disease events (both fatal and nonfatal); quality of life (Short Form-36); and mood (Hospital Anxiety and Depression Scale). MI did not significantly change measures of blood pressure (mean difference in change, -0.2.35 [95% confidence interval, -6.16 to 1.47]) or cholesterol (mean difference in change, -0.0.12 [95% confidence interval, -0.30 to 0.06]). However, it had positive effects on self-reported medication adherence at 6 months (1.979; 95% confidence interval, 0.98-3.98; P=0.0557) and 9 months (4.295; 95% confidence interval, 1.56-11.84; P=0.0049) post stroke. Improvement across other measures was also observed, but the differences between MI and usual care groups were not statistically significant. MI improved self-reported medication adherence. All other effects were nonsignificant, though in the direction of a treatment effect. Further study is required to determine whether MI leads to improvement in other important areas of functioning (eg, caregiver burden). URL: http://www.anzctr.org.au. Unique identifier: ACTRN-12610000715077.


Increased mammography use and its impact on earlier breast cancer detection in Vermont, 1975-1999.
A trend toward earlier breast carcinoma detection in the United States has been attributed to screening mammography, although direct evidence linking this trend to the increased use of mammography in a general population is lacking. This study examined the effects of mammography on tumor size and axillary lymph node metastasis in Vermont over 25 years. Pathology and mammography data from 3499 Vermont women who were diagnosed with invasive breast carcinoma during 1975-1984, 1989-1990, and 1995-1999 were compared. Logistic regression analysis was used to estimate the effects of age, mammography use, and period on the odds of a tumor < or = 2 cm and the odds of negative lymph nodes. The proportion of breast tumors that were detected by screening mammography increased from 2% during 1974-1984 to 36% during 1995-1999 (P < 0.001), and these tumors were more likely to measure < or = 2 cm than tumors that were detected by other methods. Among women age > 50 years, the odds ratio (OR) was 4.5, with a 95% confidence interval (95% CI) of 3.5-6.4. The effect was smaller in younger women (OR, 1.8; 95% CI, 1.1-3.0). Mammographic detection increased the odds of negative lymph nodes by a similar amount in both age groups, although women age > 50 years were more likely to have negative lymph nodes than younger women (OR, 1.3; 95% CI, 1.1-1.6). Tumor size and lymph node metastasis also were related to the number of mammograms and to the mammographic interval. Most of the trend toward earlier detection in Vermont was due to mammography. Mammography had a lesser effect on tumor size among younger women, which may be related to less frequent screening, although its effect on lymph node metastasis was not age dependent. Women age < 50 years were more likely to have positive lymph nodes, independent of the method of detection or the frequency of mammography.


Challenges in control of COVID-19: short doubling time and long delay to effect of interventions.
Early assessments of the growth rate of COVID-19 were subject to significant uncertainty, as expected with limited data and difficulties in case ascertainment, but as cases were recorded in multiple countries, more robust inferences could be made. Using multiple countries, data streams and methods, we estimated that, when unconstrained, European COVID-19 confirmed cases doubled on average every 3 days (range 2.2-4.3 days) and Italian hospital and intensive care unit admissions every 2-3 days; values that are significantly lower than the 5-7 days dominating the early published literature. Furthermore, we showed that the impact of physical distancing interventions was typically not seen until at least 9 days after implementation, during which time confirmed cases could grow eightfold. We argue that such temporal patterns are more critical than precise estimates of the time-insensitive basic reproduction number R0 for initiating interventions, and that the combination of fast growth and long detection delays explains the struggle in countries' outbreak response better than large values of R0 alone. One year on from first reporting these results, reproduction numbers continue to dominate the media and public discourse, but robust estimates of unconstrained growth remain essential for planning worst-case scenarios, and detection delays are still key in informing the relaxation and re-implementation of interventions. This article is part of the theme issue 'Modelling that shaped the early COVID-19 pandemic response in the UK'.


Ocular sequelae in premature infants.
The authors report on the incidence of myopia and strabismus at 12 and 24 months postterm in a cohort of 190 premature infants with birth weights of less than 1251 g born in 1986 and 1987. The neonatal and follow-up eye charts of a cohort of 190 premature infants were retrospectively reviewed. All 138 children who survived the neonatal period had at least one eye examination between day 28 and 42 of life that documented the presence and staging of retinopathy of prematurity (ROP) according to the International Classification of ROP. No infants received cryotherapy. Eye examinations conducted at 12 and 24 months postterm included assessment of vision, fundus, ocular motility, anterior segment abnormality, and refractive error. Eyes were refracted using cycloplegic retinoscopy. Strabismus was detected using the Hirschberg and cover tests. Eye reports were available for 80% (n = 110) at 12 months and 36% (n = 50) at 24 months. Fifty-three percent of the cohort exhibited ROP in the neonatal period; 12% of these progressed to stage 3 or 4 ROP. Myopia was observed in 16% (18/110) of the cohort at 12 months of age; 4.5% (5/110) measured more than 4.0 diopters of myopia. Children with birth weights of less than 751 g were 3.2 times more likely than those with birth weights between 751 and 1000 g and 10 times more likely than those with birth weights between 1001 and 1250 g to develop myopia in the first year of life. The likelihood of myopia at 12 months doubled with each increment in ROP stage. Of the 50 children reexamined at 24 months postterm, more than 80% demonstrated deteriorating vision. The incidence of myopia increased to 38% (19/50) overall, with 24% (12/50) of the cohort showing severe myopia. Astigmatism and anisometropia were highly correlated with severe myopia. Strabismus was seen with increasing frequency through the second year of life. All children with grade III or IV intraventricular hemorrhage in the neonatal period developed esotropia. This study emphasizes the significant roles of low birth weight, ROP, and intraventricular hemorrhage in the development of myopia and strabismus. Follow-up to 2 years of life is recommended given the demonstrated deterioration in our cohort.


Correlation among Metabolic Changes in Tea Plant Camellia sinensis (L.) Shoots, Green Tea Quality and the Application of Cow Manure to Tea Plantation Soils.
Traditionally, the supplement of organic manure in tea plantations has been a common approach to improving soil fertility and promoting terroir compounds, as manifested by the coordinated increase in yield and quality for the resulting teas. However, information regarding the effect of organic manure in the metabolome of tea plants is still inadequate. The metabolite profiles of tea shoots applied with cow manure, urea or no fertilizer were studied using gas chromatography-mass spectrometry (GC-MS). In total, 73 metabolites were detected, and the modulated metabolites included mainly amino acids, organic acids and fatty acids. In particular, glutamine, quinic acid and proline accumulated more in tea shoots in soils treated with cow manure, but octadecanoic acid, hexadecanoic acid and eicosanoic acid were drastically reduced. Pearson correlation analysis indicated that organic acids and amino acids in tea shoots were the two major metabolite groups among the three treatments. The analysis of metabolic pathways demonstrated that the cow manure treatment significantly changed the enrichment of pathways related to amino acids, sugars and fatty acids. Sensory evaluation showed that the quality of green teas was higher when the plants used to make the tea were grown in soil treated with cow manure rather than urea during spring and late summer. The results indicated that the application of cow manure in soils changed the metabolic characteristics of tea shoots and improved the qualities of the resulting teas.


Randomized Study of Maintenance Pemetrexed Versus Observation for Treatment of Malignant Pleural Mesothelioma: CALGB 30901.
The role of maintenance therapy for malignant pleural mesothelioma (MPM) is unknown. We performed a randomized phase II trial to determine if continuation of pemetrexed after first-line pemetrexed and platinum would improve progression-free survival (PFS). Eligible patients with unresectable MPM, without disease progression following 4 to 6 cycles of pemetrexed and platinum were randomized 1:1 to observation or continuation of pemetrexed until progression, stratified by number of cycles (< 6 or 6), cis- or carboplatin containing regimen, and histology. Study size was calculated based on the assumption that observation would produce a median PFS of 3 months and pemetrexed would yield median PFS of 6 months. A total of 72 patients were registered from December 2010 to June 2016. The study closed early after 53 patients were randomized; 49 eligible (22 on the observation arm and 27 on the pemetrexed arm) were included in the analysis. The median PFS was 3 months (95% confidence interval [CI], 2.6-11.9 months) on observation and 3.4 months (95% CI, 2.8-9.8 months) on pemetrexed (hazard ratio [HR], 0.99; 95% CI, 0.51-1.90; P = .9733). The median overall survival (OS) was 11.8 months (95% CI, 9.3-28.7 months) for observation, and 16.3 months (95% CI, 10.5-26.0 months) for pemetrexed (HR, 0.86; 95% CI, 0.44-1.71; P = .6737). Grade 3 or 4 toxicities on the pemetrexed arm included anemia (8%), lymphopenia (8%), neutropenia (4%), and fatigue (4%). A higher baseline level of soluble mesothelin-related peptide was associated with worse PFS (HR, 1.86; 95% CI, 1.00-3.46; P = .049). Maintenance pemetrexed following initial pemetrexed and platinum chemotherapy does not improve PFS in patients with MPM.


Organics filled one-dimensional TiO2 nanowires array ultraviolet detector with enhanced photo-conductivity and dark-resistivity.
A heterojunction photo-conductive ultraviolet (UV) detector was developed based on TiO2 nanowires array (NWA) surrounded by N,N'-bis-(1-naphthalenyl)-N,N'-bis-phenyl-(1,1'-biphenyl)-4,4'-diamine (NPB). The novel and effective two-step method of static infusion and dynamic solution-cleaning was employed to fill NPB into TiO2 NWA gaps and simultaneously remove the unwelcomed top NPB layer. The device fabricated via the two-step method exhibited optimal performance compared to TiO2/NPB device with top NPB layer and TiO2 NWA device. In dark conditions, the TiO2/NPB heterojunction device without top NPB was found to possess the capacity of depleting majority carriers, thereby providing improved dark-resistivity to limit the dark current (Id). Under UV illumination, the depleting effect could be eliminated by the dissociation and accumulation of photo-generated carriers between pn heterojunction, leading to increased carrier density and photo-conductivity. It cleared up the high barrier due to the removal of top NPB layer, which was beneficial for hot electron transport than the device with top NPB layer under illumination, thereby achieving an enhanced light current (Il) to Id ratio of 1.67 × 104. A simple technology is provided to prepare organic-inorganic hybrid one-dimensional array heterostructure, which plays a remarkable role in the working of the UV detector, enhancing photo-conductivity and dark-resistivity of the device.


The asthma/COPD nurses' experience of educating patients with chronic obstructive pulmonary disease in primary health care.
The number of patients with chronic obstructive pulmonary disease (COPD) is increasing. These patients need nursing care, including education in self-care, which has a positive effect on their physical and psychoemotional well-being. The aim of this study was to describe the experiences of asthma/COPD nurses' in primary health care (PHC) of educating patients with COPD. A descriptive, qualitative study was conducted, with interviews of 12 asthma/COPD nurses. The data were analysed using qualitative content analysis. The findings are presented in two themes: Theme 1, receiving support results in a feeling of security, which enables the development of patient education; and Theme 2, a lack of support results in a feeling of insecurity, which makes it difficult to develop patient education. The asthma/COPD nurses were individual orientated with individualization of care, but the patient's mood, the varying support of those around and the nurses' varying degrees of security affected the education. The conclusion is that the asthma/COPD nurses' experience of patient education fluctuated between insecurity and security. The nurses' feeling of insecurity in their patient education can be strengthened through support from colleagues and by increased knowledge in promoting the learning of others. Collaborative teamwork with a well-functioning asthma/COPD clinic in PHC can facilitate and improve patient services; these initiatives can enable the asthma/COPD nurses to reach their full potential.


Predictive and prognostic value of stromal tumour-infiltrating lymphocytes before and after neoadjuvant therapy in triple negative and HER2-positive breast cancer.
Lymphocyte predominant breast cancer (BC) is associated with higher pathological complete response (pCR) rate after neoadjuvant therapy (NAT) and favorable outcome in triple negative breast cancer (TNBC) and HER2+ BC. The predictive and prognostic impact of stromal tumour-infiltrating lymphocytes (TILs) after NAT and the change of TILs before (pre-) and after (post-) NAT are not well studied. We aimed to assess the predictive and prognostic value of pre- and post-NAT TILs, as well as their pharmacodynamics modulation and their change for TNBC and HER2+ BC. Two-hundred and nine consecutive patients (n = 80 TNBC, n = 129 HER2+ BC) who received NAT between 2001 and 2009 in a single institution were included. We evaluated the association between pre-NAT TILs and pCR, and the association between pre- and post-NAT TILs, as well as their immunodynamics change with relapse-free survival (RFS) for patients with residual disease (RD). Low pre-NAT TILs compared to int/high were significantly associated with lower pCR rate (TNBC: 4.0% vs 43.6%; HER2+ BC: 26.0% vs 51.9%). The median follow-up period was 98 months. In TNBC with RD, low pre-NAT TILs showed significant association with shorter RFS (HR = 3.844 [1.190-12.421], p = 0.024) in multivariate analysis. Low post-NAT TILs showed borderline significant association with shorter RFS (HR = 2.836 [0.951-8.457], p = 0.061). The change in TILs was not associated with RFS. In HER2+ BC, low pre-NAT TILs were not associated with RFS. In TN and HER2+ BCs, low pre-NAT TILs tumours had a low likelihood of achieving pCR. In TNBC with RD, both low pre- and post-NAT TILs were associated with shorter RFS. These results suggest that TILs information should be taken into account when additional therapies may be given in the post-neoadjuvant setting.


Interactions between small heat shock protein subunits and substrate in small heat shock protein-substrate complexes.
Small heat shock proteins (sHSPs) are dynamic oligomeric proteins that bind unfolding proteins and protect them from irreversible aggregation. This binding results in the formation of sHSP-substrate complexes from which substrate can later be refolded. Interactions between sHSP and substrate in sHSP-substrate complexes and the mechanism by which substrate is transferred to ATP-dependent chaperones for refolding are poorly defined. We have established C-terminal affinity-tagged sHSPs from a eukaryote (pea HSP18.1) and a prokaryote (Synechocystis HSP16.6) as tools to investigate these issues. We demonstrate that sHSP subunit exchange for HSP18.1 and HSP16.6 is temperature-dependent and rapid at the optimal growth temperature for the organism of origin. Increasing the ratio of sHSP to substrate during substrate denaturation decreased sHSP-substrate complex size, and accordingly, addition of substrate to pre-formed sHSP-substrate complexes increased complex size. However, the size of pre-formed sHSP-substrate complexes could not be reduced by addition of more sHSP, and substrate could not be observed to transfer to added sHSP, although added sHSP subunits continued to exchange with subunits in sHSP-substrate complexes. Thus, although some number of sHSP subunits within complexes remain dynamic and may be important for complex structure/solubility, association of substrate with the sHSP does not appear to be similarly dynamic. These observations are consistent with a model in which ATP-dependent chaperones associate directly with sHSP-bound substrate to initiate refolding.


Occurrence of malignant vertebral fractures in an emergency room setting.
To perform a risk analysis study to determine the probability of a spinal fracture being of malignant origin in patients presenting at a level I trauma centre emergency room after trauma. Data from 334 consecutive patients were retrospectively obtained from 1993 to 2003. They were divided into two groups: group 1--(benign) traumatic fractures; and group 2--malignant fractures (n = 32). For statistical analysis independent Student t test, chi2 test, and backward-stepwise logistic regression were used. The risk of vertebral fractures appearing to be of malignant origin increased with anatomical location (non-cervical--that is, thoracic or lumbar: odds ratio (OR) 48, 95% confidence interval (CI) 8 to 291), a history of malignancy (OR 72, 95% CI 12 to 422), trauma mechanism (that is, high energy: OR 0.03, 95% CI 0.003 to 0.28), and age >64 years (OR 3, 95% CI 0.9 to 12). Hence, patients over 64 years old attending the emergency room, with a vertebral fracture after a low energy trauma, had an approximately 50% chance of having a malignant fracture. With a non-cervical location and a history of malignancy this increased to 98%. Regardless of the trauma mechanism and age of the patient, a history of a malignancy and a non-cervical fracture posed at least a 36% risk of having a malignant fracture. Supported by the present results we feel the probability of malignant fractures, although not frequently encountered, should always be considered in elderly and middle-aged patients with a history of malignancy and a non-cervical traumatic fracture.


An integrin phosphorylation switch: the effect of beta3 integrin tail phosphorylation on Dok1 and talin binding.
Integrins play a fundamental role in cell migration and adhesion; knowledge of how they are regulated and controlled is vital for understanding these processes. Recent work showed that Dok1 negatively regulates integrin activation, presumably by competition with talin. To understand how this occurs, we used NMR spectroscopy and x-ray crystallography to investigate the molecular details of interactions with integrins. The binding affinities of beta3 integrin tails for the Dok1 and talin phosphotyrosine binding domains were quantified using 15N-1H hetero-nuclear single quantum correlation titrations, revealing that the unphosphorylated integrin tail binds more strongly to talin than Dok1. Chemical shift mapping showed that unlike talin, Dok1 exclusively interacts with the canonical NPXY motif of the beta3 integrin tail. Upon phosphorylation of Tyr 747 in the beta3 integrin tail, however, Dok1 then binds much more strongly than talin. Thus, we show that phosphorylation of Tyr 747 provides a switch for integrin ligand binding. This switch may represent an in vivo mechanism for control of integrin receptor activation. These results have implications for the control of integrin signaling by proteins containing phosphotyrosine binding domains.


Repeated salivary daytime cortisol and onset of mood episodes in offspring of bipolar parents.
Differences in cortisol secretion may differentiate individuals at high compared to low genetic risk for bipolar disorder (BD) and predict the onset or recurrence of mood episodes. The objectives of this study were to determine if salivary cortisol measures are: (1) different in high-risk offspring of parents with BD (HR) compared to control offspring of unaffected parents (C), (2) stable over time, (3) associated with the development of mood episode onset/recurrence, and (4) influenced by comorbid complications. Fifty-three HR and 22 C completed salivary cortisol sampling annually for up to 4 years in conjunction with semi-structured clinical interviews. The cortisol awakening response (CAR), daytime cortisol [area under the curve (AUC)], and evening cortisol (8:00 p.m.) were calculated. There were no differences in baseline CAR, AUC and evening cortisol between HR and C (p = 0.38, p = 0.30 and p = 0.84), respectively. CAR, AUC and evening cortisol were stable over yearly assessments in HR, while in Cs, evening cortisol increased over time (p = 0.008), and CAR and AUC remained stable. In HR, AUC and evening cortisol increased the hazard of a new onset mood disorder/recurrence by 2.7 times (p = 0.01), and 3.5 times (p = 0.01), respectively, but this was no longer significant after accounting for multiple comparisons. Salivary cortisol is stable over time within HR offspring. However, between individuals, basal salivary cortisol is highly variable. More research is needed, with larger samples of prospectively studied HR youth using a more reliable method of cortisol measurement, to determine the potential role of cortisol in the development of mood disorders.


Experimental Support That Natural Selection Has Shaped the Latitudinal Distribution of Mitochondrial Haplotypes in Australian Drosophila melanogaster.
Cellular metabolism is regulated by enzyme complexes within the mitochondrion, the function of which are sensitive to the prevailing temperature. Such thermal sensitivity, coupled with the observation that population frequencies of mitochondrial haplotypes tend to associate with latitude, altitude, or climatic regions across species distributions, led to the hypothesis that thermal selection has played a role in shaping standing variation in the mitochondrial DNA (mtDNA) sequence. This hypothesis, however, remains controversial, and requires evidence that the distribution of haplotypes observed in nature corresponds with the capacity of these haplotypes to confer differences in thermal tolerance. Specifically, haplotypes predominating in tropical climates are predicted to encode increased tolerance to heat stress, but decreased tolerance to cold stress. We present direct evidence for these predictions, using mtDNA haplotypes sampled from the Australian distribution of Drosophila melanogaster. We show that the ability of flies to tolerate extreme thermal challenges is affected by sequence variation across mtDNA haplotypes, and that the thermal performance associated with each haplotype corresponds with its latitudinal prevalence. The haplotype that predominates at low (subtropical) latitudes confers greater resilience to heat stress, but lower resilience to cold stress, than haplotypes predominating at higher (temperate) latitudes. We explore molecular mechanisms that might underlie these responses, presenting evidence that the effects are in part regulated by SNPs that do not change the protein sequence. Our findings suggest that standing variation in the mitochondrial genome can be shaped by thermal selection, and could therefore contribute to evolutionary adaptation under climatic stress.


Ozone as u-shaped dose responses molecules (hormetins).
Redox environment involves a broad network of pro-oxidant and antioxidant components. Health benefit or damage can be induced as a consequence of an adaptive cellular stress response. A consequence of hormetic amplification is an increase in the homeodynamic space of a living system in terms of an increased defense capacity and a reduced load of damaged macromolecules. Ozone, when used at appropriate doses, promotes the formation of reactive oxygen species and lipid peroxides allows them to become late and long-lasting messengers. Healthy aging may be achieved by hormesis through mild and periodic, but not severe or chronic, physical and mental challenges, and by the use of nutritional hormesis incorporating mild stress-inducing molecules called hormetins. The paradoxical concept that ozone eventually induces an antioxidant response capable of reversing a chronic oxidative stress is common in the animal and vegetal kingdom; it is already supported by findings of an increased level of antioxidant enzymes during ozone therapy. Those facts can include ozone as a hormetin. The established scientific foundations of hormesis are ready to pave the way for new and effective approaches in redox-related disease research and intervention; ozone therapy can be a good candidate.


Monitoring physical and psychosocial symptom trajectories in ovarian cancer patients receiving chemotherapy.
Diagnosis and treatment of ovarian cancer (OC) entail severe symptom burden and a significant loss of quality of life (QOL). Somatic and psychological impairments may persist well beyond active therapy. Although essential for optimal symptom management as well as for the interpretation of treatment outcomes, knowledge on the course of QOL-related issues is scarce. This study aimed at assessing the course of depressive symptoms, anxiety, fatigue and QOL in patients with OC over the course of chemotherapy until early after-care. 23 patients were assessed longitudinally (eight time points) with regard to symptom burden (depression, anxiety, fatigue, and QOL) by means of patient-reported outcome instruments (HADS, MFI-20, EORTC QLQ-C30/-OV28) and clinician ratings (HAMA/D) at each chemotherapy cycle and at the first two aftercare visits. Statistically significant decrease over time was found for depressive symptoms and anxiety as well as for all fatigue scales. With regard to QOL, results indicated significant increase for 11 of 15 QOL scales, best for Social (effect size = 1.95; p < 0.001), Emotional (e.s. = 1.62; p < 0.001) and Physical Functioning (e.s. = 1.47; p < 0.001). Abdominal Symptoms (e.s. = 1.01; p = 0.009) decreased, Attitudes towards Disease and Treatment (e.s. = 1.80; p < 0.001) improved significantly over time. Analysis of Sexual Functioning was not possible due to a high percentage of missing responses (61.9%). The present study underlines the importance of longitudinal assessment of QOL in order to facilitate the identification of symptom burden in OC patients. We found that patients show high levels of fatigue, anxiety and depressive symptoms and severely impaired QOL post-surgery (i.e. at start of chemotherapy) but condition improves considerably throughout chemotherapy reaching nearly general population symptoms levels until aftercare.


Evolution and developmental expression of the sodium-iodide symporter (NIS, slc5a5) gene family: Implications for perchlorate toxicology.
The vertebrate sodium-iodide symporter (NIS or SLC5A5) transports iodide into the thyroid follicular cells that synthesize thyroid hormone. The SLC5A protein family includes transporters of vitamins, minerals, and nutrients. Disruption of SLC5A5 function by perchlorate, a pervasive environmental contaminant, leads to human pathologies, especially hypothyroidism. Perchlorate also disrupts the sexual development of model animals, including threespine stickleback (Gasterosteus aculeatus) and zebrafish (Danio rerio), but the mechanism of action is unknown. To test the hypothesis that SLC5A5 paralogs are expressed in tissues necessary for the development of reproductive organs, and therefore are plausible candidates to mediate the effects of perchlorate on sexual development, we first investigated the evolutionary history of Slc5a paralogs to better understand potential functional trajectories of the gene family. We identified two clades of slc5a paralogs with respect to an outgroup of sodium/choline cotransporters (slc5a7); these clades are the NIS clade of sodium/iodide and lactate cotransporters (slc5a5, slc5a6, slc5a8, slc5a8, and slc5a12) and the SGLT clade of sodium/glucose cotransporters (slc5a1, slc5a2, slc5a3, slc5a4, slc5a10, and slc5a11). We also characterized expression patterns of slc5a genes during development. Stickleback embryos and early larvae expressed NIS clade genes in connective tissue, cartilage, teeth, and thyroid. Stickleback males and females expressed slc5a5 and its paralogs in gonads. Single-cell transcriptomics (scRNA-seq) on zebrafish sex-genotyped gonads revealed that NIS clade-expressing cells included germ cells (slc5a5, slc5a6a, and slc5a6b) and gonadal soma cells (slc5a8l). These results are consistent with the hypothesis that perchlorate exerts its effects on sexual development by interacting with slc5a5 or its paralogs in reproductive tissues. These findings show novel expression domains of slc5 genes in stickleback and zebrafish, which suggest similar functions across vertebrates including humans, and provide candidates to mediate the effects of perchlorate on sexual development.


Turkish Urologists' preferences regarding antibiotic prophylaxis for transrectal prostate biopsy.
Antibiotic prophylaxis for transrectal prostate biopsy (PB) is very important in preventing infectious complications, and in this study, we aimed to evaluate the antibiotic preferences of Turkish urologist for transrectal PB. The survey about PB and antibiotic prophylaxis behaviors was administered to urologists working in Turkey who had previously participated in at least one international and one national congress. A total of 237 urologists were included in the study. Antibiotic prophylaxis prior to PB was performed by 234 (98.7%) participants. Rectal swabbing prior to PB was not performed by 227 (95.8%) participants. The most common complication associated with PB was prostatitis (63%), followed by urinary tract infection (29%). Only 25.7% of Turkish urologists reported a single dose of antibiotic prophylaxis. Our participants often administered antibiotic prophylaxis for a period of 3, 5, or 7 days (16%, 21.1%, 35.9%, respectively). The most common antibiotic agent preferred for prophylaxis was ciprofloxacin (65%). The biopsy behavior of Turkish urologists was mostly compatible with the literature, but it was revealed that Turkish urologists do not prefer single-dose antibiotic therapy, and their practice patterns regarding the administration of pre-biopsy rectal swabbing are inconsistent with the literature.


Dietary Intake Is Associated with Occlusal Force Rather Than Number of Teeth in 80-y-Old Japanese.
There has been a growing interest in the association between the number of teeth and dietary intake in older populations. However, people around the age of 80 y have frequently lost most of their teeth, and dental prostheses replacing the missing teeth play an important role in masticatory function. Therefore, masticatory function cannot be evaluated by the number of teeth alone. The occlusal force of the complete dental arches is an index of masticatory function, reflecting not only the number of teeth, but the effect of removable dentures. The purpose of this cross-sectional study was to determine the relative importance of the number of teeth and occlusal force in association with dietary intake in 80-y-old Japanese people. This study included 760 community-dwelling Japanese people aged 79 y to 81 y. The authors measured bilateral maximal occlusal force in the intercuspal position using pressure-sensitive sheets. Removable denture wearers kept their dentures in place during the measurements. Energy-adjusted food groups and nutrient intake during the preceding month were assessed by a brief self-administered diet history questionnaire. The authors assessed linear trends in food and nutrient intake in relation to the number of teeth and occlusal force after adjusting for gender and socioeconomic status (education level, financial status, family structure, resident area and BMI). P values of < 0.05 were considered to be statistically significant. The authors found that the number of teeth was not associated with the energy-adjusted intake of any food group examined. In contrast, a decline in occlusal force was significantly associated with a lower intake of vegetables, fish and shellfish, protein, polyunsaturated fatty acids, dietary fiber and most vitamins and minerals ( P for trend < 0.05). We conclude that food and nutrient intake was more closely associated with occlusal force than the number of teeth in community-dwelling Japanese people aged 79 y to 81 y. Knowledge Transfer Statement: This cross-sectional study of older Japanese people showed that, after controlling for considerable covariates, occlusal force rather than the number of teeth is positively associated with energy-adjusted intake of vegetables, fish and shellfish, protein, polyunsaturated fatty acids, dietary fiber and most of vitamins and minerals. This means that reduced occlusal force may unconsciously lead older people toward a habitual unhealthy dietary intake. Older people have frequently lost most of their teeth and require prosthetics to restore masticatory function. Bilateral occlusal force is therefore a better measure of masticatory function than the number of remaining teeth. Our findings suggest that prosthetic rehabilitation is a significant factor in the prevention and management of chronic diseases and frailty through better dietary intake in older populations.


A prediction model for peak power output from different incremental exercise tests.
There is currently no model that predicts peak power output (PPO) thereby allowing comparison between different incremental exercise test (EXT) protocols. In this study we have used the critical power profile to develop a mathematical model for predicting PPO from the results of different EXTs. The purpose of this study was to examine the level of agreement between actual PPO values and those predicted from the new model. Eleven male athletes (age 25 +/- 5 years, VO(2max) 62 +/- 8 mL . kg(-1) . min(-1)) completed 3 laboratory tests on a cycle ergometer. Each test comprised an EXT consisting of 1-min workload increments of 30 W (EXT30/1) and 3-min (EXT25/3) and 5-min workload increments (EXT25/5) of 25 W. The PPO determined from each test was used to predict the PPO from the remaining 2 EXTs. The differences between actual and predicted PPO values were statistically insignificant (P > .05). The random error components of the limits of agreement of <or=30 W also indicated acceptable levels of agreement between actual and predicted PPO values. Further data collection is necessary to confirm whether the model is able to predict PPO over a wide range of EXT protocols in athletes of different aerobic and anaerobic capacities.


Safety and reactogenicity of canarypox ALVAC-HIV (vCP1521) and HIV-1 gp120 AIDSVAX B/E vaccination in an efficacy trial in Thailand.
A prime-boost vaccination regimen with ALVAC-HIV (vCP1521) administered intramuscularly at 0, 4, 12, and 24 weeks and gp120 AIDSVAX B/E at 12 and 24 weeks demonstrated modest efficacy of 31.2% for prevention of HIV acquisition in HIV-uninfected adults participating in a community-based efficacy trial in Thailand. Reactogenicity was recorded for 3 days following vaccination. Adverse events were monitored every 6 months for 3.5 years, during which pregnancy outcomes were recorded. Of the 16,402 volunteers, 69% of the participants reported an adverse event any time after the first dose. Only 32.9% experienced an AE within 30 days following any vaccination. Overall adverse event rates and attribution of relatedness did not differ between groups. The frequency of serious adverse events was similar in vaccine (14.3%) and placebo (14.9%) recipients (p = 0.33). None of the 160 deaths (85 in vaccine and 75 in placebo recipients, p = 0.43) was assessed as related to vaccine. The most common cause of death was trauma or traffic accident. Approximately 30% of female participants reported a pregnancy during the study. Abnormal pregnancy outcomes were experienced in 17.1% of vaccine and 14.6% (p = 0.13) of placebo recipients. When the conception occurred within 3 months (estimated) of a vaccination, the majority of these abnormal outcomes were spontaneous or elective abortions among 22.2% and 15.3% of vaccine and placebo pregnant recipients, respectively (p = 0.08). Local reactions occurred in 88.0% of vaccine and 61.0% of placebo recipients (p<0.001) and were more frequent after ALVAC-HIV than AIDSVAX B/E vaccination. Systemic reactions were more frequent in vaccine than placebo recipients (77.2% vs. 59.8%, p<0.001). Local and systemic reactions were mostly mild to moderate, resolving within 3 days. The ALVAC-HIV and AIDSVAX B/E vaccine regimen was found to be safe, well tolerated and suitable for potential large-scale use in Thailand. ClinicalTrials.govNCT00223080.


How are non-numerical prognostic statements interpreted and are they subject to positive bias?
Frank, clear communication with family members of terminally ill or incapacitated patients has important implications for well-being, satisfaction with care and sound decision-making. However, numerical prognostic statements, particularly more negative ones, have been found to be interpreted in a positively biased manner. Less precise non-numerical statements, preferred by physicians, and particularly statements using threatening terms (dying vs surviving) may be even more subject to such biases. Participants (N=200) read non-numerical prognostic statements framed in terms of dying or surviving and indicated their interpretation of likelihood of survival. Even the most extreme statements were not interpreted to indicate 100% likelihood of surviving or dying, (eg, they will definitely survive, 92.77%). The poorness of prognoses was associated with more optimistically biased interpretations but this was not, however, affected by the wording of the prognoses in terms of dying versus surviving. The findings illuminate the ways in which commonly used non-numeric language may be understood in numeric terms during prognostic discussions and provide further evidence of recipients' propensity for positive bias.


Self-disclosure by adolescents in therapy for eating difficulties: A Q-Methodology study.
Facilitating client self-disclosure is essential to therapeutic effectiveness. Given the long-term consequences of eating difficulties for adolescents, there is a need for more research on self-disclosure in this group. This study investigated factors likely to influence adolescents' decisions to self-disclose during psychological therapy for eating difficulties using Q-methodology. Participants (n = 28), recruited through child and adolescent mental health services in the UK, completed a task that involved sorting 47 statements to represent their viewpoint on self-disclosure. The 28 completed sorts were subjected to a by-participant factor analysis in order to identify distinct viewpoints in the sample. Three distinct factors were extracted. One factor emphasised the importance of therapist self-disclosure on decisions to disclose. In contrast, another factor placed more emphasis on the influence of eating disorder identity and readiness to change on disclosure decisions. The third factor placed emphasis on the quality of the therapeutic relationship and readiness to change as having most influence. Given the absence of a unifying factor representing what influences the decision to disclose, clinicians should ensure they explore with young people what might influence their decision to disclose.


A Step-by-Step Guide to Assemble a Reptilian Genome.
Multiple technologies and software are now available facilitating the de novo sequencing and assembly of any vertebrate genome. Yet the quality of most available sequenced genomes is substantially poorer than that of the golden standard in the field: the human genome. Here, we present a step-by-step protocol for the successful sequencing and assembly of a high-quality snake genome that can be applied to any other reptilian or avian species. We combine the great sequencing depth and accuracy of short reads with the use of different insert size libraries for extended scaffolding followed by optical mapping. We show that this procedure improved the corn snake scaffold N50 from 3.7 kbp to 1.4 Mbp, currently making it one of the snake genomes with the longest scaffolds. Short guidelines are also given on the extraction of long DNA molecules from reptilian blood and the necessary modifications in DNA extraction protocols. This chapter is accompanied by a website ( www.reptilomics.org/stepbystep.html ), where we provide links to the suggested software, examples of input and output files, and running parameters.


Moraxella catarrhalis M35 is a general porin that is important for growth under nutrient-limiting conditions and in the nasopharynges of mice.
Moraxella catarrhalis is a gram-negative respiratory pathogen that is an important causative agent for otitis media and exacerbations of chronic obstructive pulmonary disease. We have previously predicted the outer membrane protein M35 to be a general porin, and in the current study, we have investigated the function of M35 and its importance for survival of M. catarrhalis in vivo. Lipid bilayer experiments reveal that refolded M35 functions as a channel that is typical of gram-negative bacterial porins. M35 forms wide and water-filled channels with a single-channel conductance of about 1.25 nS in 1 M KCl solution and has only a small selectivity for cations over anions. When the in vitro growth characteristics of two M35 deletion mutant strains of M. catarrhalis were compared to the wild-type parent isolates, the growth of the mutant strains was inhibited only under nutrient-poor conditions. This growth defect could be eliminated by additional glutamic acid, but not additional aspartic acid, glycine, sucrose, or glucose. The mutant strains compensated for the lack of M35 by enhancing their uptake of glutamic acid, and this enhanced rate of glutamic acid uptake was attributed to the compensatory upregulation of a protein of approximately 40 kDa. M35 was also found to be essential for nasal colonization of mice, demonstrating that its presence is essential for survival of M. catarrhalis in vivo. These results suggest that M35 is a general porin that is necessary for the uptake of important energy sources by M. catarrhalis and that it is likely that M35 is an essential functional protein for in vivo colonization.


Brands, costs and registration status of antimalarial drugs in the Kenyan retail sector.
Although an important source of treatment for fevers, little is known about the structure of the retail sector in Africa with regard to antimalarial drugs. This study aimed to assess the range, costs, sources and registration of antimalarial drugs in the Kenyan retail sector. In 2002, antimalarial drug registration and trade prices were established by triangulating national registration lists, government gazettes and trade price indices. Data on registration status and trade prices were compared with similar data generated through a retail audit undertaken among 880 randomly sampled retailers in four districts of Kenya. Two hundred and eighteen antimalarial drugs were in circulation in Kenya in 2002. These included 65 "sulfur"-pyrimethamine (sulfadoxine-pyrimethamine and sulfalene-pyrimethamine (SP), the first-line recommended drug in 2002) and 33 amodiaquine (AQ, the second-line recommended drug) preparations. Only half of SP and AQ products were registered with the Pharmacy and Poisons Board. Of SP and AQ brands at district level, 40% and 44% were officially within legal registration requirements. 29% of retailers at district level stocked SP and 95% stocked AQ. The retail price of adult doses of SP and AQ were on average 0.38 and 0.76 US dollars, 100% and 347% higher than trade prices from manufacturers and importers. Artemether-lumefantrine, the newly announced first-line recommended antimalarial drug in 2004, was found in less than 1% of all retail outlets at a median cost of 7.6 US dollars. There is a need to ensure that all antimalarial drugs are registered with the Pharmacy and Poisons Board to facilitate a more stringent post-marketing surveillance system to ensure drugs are safe and of good quality post-registration.


In vitro reconstitution of an mRNA-transport complex reveals mechanisms of assembly and motor activation.
The assembly and composition of ribonucleic acid (RNA)-transporting particles for asymmetric messenger RNA (mRNA) localization is not well understood. During mitosis of budding yeast, the Swi5p-dependent HO expression (SHE) complex transports a set of mRNAs into the daughter cell. We recombinantly reconstituted the core SHE complex and assessed its properties. The cytoplasmic precomplex contains only one motor and is unable to support continuous transport. However, a defined interaction with a second, RNA-bound precomplex after its nuclear export dimerizes the motor and activates processive RNA transport. The run length observed in vitro is compatible with long-distance transport in vivo. Surprisingly, SHE complexes that either contain or lack RNA cargo show similar motility properties, demonstrating that the RNA-binding protein and not its cargo activates motility. We further show that SHE complexes have a defined size but multimerize into variable particles upon binding of RNAs with multiple localization elements. Based on these findings, we provide an estimate of number, size, and composition of such multimeric SHE particles in the cell.


Inter-relatedness of underlying factors for injury and violence among adolescents in rural coastal Kenya: A qualitative study.
We utilized a socio-ecological model to explore views from 85 young people and 10 local stakeholders on forms and underlying factors for unintentional injury, violence, self-harm, and suicidal behavior of adolescents in Kilifi County, Kenya. Young people took part in 11 focus group discussions, whereas 10 in-depth interviews were conducted with the local stakeholders. Road traffic accidents, falls, fights, sexual and gender-based violence, theft, and vandalism were viewed as common. There was an overlap of risk factors, especially at intra- and interpersonal levels (gender, poverty, substance use, parenting behavior, school drop-out). Some broader-level risk factors were insecure neighborhoods and risky sources of livelihood. Research is needed to quantify burden and to pilot feasible injury prevention interventions in this setting.


Cancer therapy utilizing an adenoviral vector expressing only E1A.
The human adenovirus type 5 (Ad5) early region 1A (E1A) proteins have been shown to have potent antitumor effects, due to their ability to reprogram oncogenic signalling pathways in tumor cells. The success of E1A antitumor therapy in animal models has led to its use in phase I and phase II clinical trials, where liposome-based delivery vehicles are being used to deliver a plasmid encoding E1A. To increase the efficiency of E1A delivery to tumors, we have developed an Ad vector deleted of all viral protein coding sequences (termed helper-dependent Ad vectors, hdAds) with the exception of E1A, designated hdAd-E1A. In culture, this vector mediated high-level expression of E1A gene products. A549 cells, a human lung adenocarcinoma cell line, infected with hdAd-E1A showed a reduced proliferative capacity in adherent culture, and the ability to form colonies in soft agarose was completely abolished. In contrast, A549 infected with an hdAd expressing beta-gal were able to form colonies of a similar size and frequency as uninfected cells. Under serum-depleted conditions, expression of E1A within A549 led to the induction of apoptosis. Finally, A549 cells treated with hdAd-E1A showed approximately 10-fold greater sensitivity to the chemotherapeutic drug cisplatin. Taken together, these data indicate that the use of hdAd provides a simple and effective method to deliver E1A to cancer cells, and results in reduction in the tumorigenic potential of the cells, as well as increasing the cells sensitivity to anticancer drugs.


Effects of high and low 17β-estradiol doses on focal cerebral ischemia: negative results.
The reasons why some animal studies indicate that estrogens increase focal cerebral ischemic damage while others show estrogen-induced neuroprotection has hitherto not been fully elucidated. Recent evidence indicates that discrepancies in hormone administration paradigms, resulting in highly different serum hormone concentrations, may account for the dichotomy. The current study aimed to test this hypothesis. Sixty ovariectomized female rats were randomized into three groups differing in 17β-estradiol regimens, and transient focal cerebral ischemia was subsequently induced. All animals were subjected to a small functional testing battery, and three days after MCAo they were sacrificed for infarct size assessment. Infarct sizes did not differ between groups, however clear discrepancies were seen in body weight and feeding behavior. In comparison to sham-operated animals, ovariectomized rats rapidly increased in body weight, whereas the opposite was seen in rats receiving 17beta-estradiol. The weight gain in the ovariectomized rats was paralleled by an increased food intake.


sp2-Iminosugar O-, S-, and N-glycosides as conformational mimics of α-linked disaccharides; implications for glycosidase inhibition.
The synthesis of mimics of the α(1→6)- and α(1→4)-linked disaccharides isomaltose and maltose featuring a bicyclic sp(2)-iminosugar nonreducing moiety O-, S-, or N-linked to a glucopyranoside residue is reported. The strong generalized anomeric effect operating in sp(2)-iminosugars determines the α-stereochemical outcome of the glycosylation reactions, independent of the presence or not of participating protecting groups and of the nature of the heteroatom. It also imparts chemical stability to the resulting aminoacetal, aminothioacetal, or gem-diamine functionalities. All the three isomaltose mimics behave as potent and very selective inhibitors of isomaltase and maltase, two α-glucosidases that bind the parent disaccharides either as substrate or inhibitor. In contrast, large differences in the inhibitory properties were observed among the maltose mimics, with the O-linked derivative being a more potent inhibitor than the N-linked analogue; the S-linked pseudodisaccharide did not inhibit either of the two target enzymes. A comparative conformational analysis based on NMR and molecular modelling revealed remarkable differences in the flexibility about the glycosidic linkage as a function of the nature of the linking atom in this series. Thus, the N-pseudodisaccharide is more rigid than the O-linked derivative, which exhibits conformational properties very similar to those of the natural maltose. The analogous pseudothiomaltoside is much more flexible than the N- or O-linked derivatives, and can access a broader area of the conformational space, which probably implies a strong entropic penalty upon binding to the enzymes. Together, the present results illustrate the importance of taking conformational aspects into consideration in the design of functional oligosaccharide mimetics.


Lysophosphatidyl Choline Induced Demyelination in Rat Probed by Relaxation along a Fictitious Field in High Rank Rotating Frame.
In this work a new MRI modality entitled Relaxation Along a Fictitious Field in the rotating frame of rank 4 (RAFF4) was evaluated in its ability to detect lower myelin content in lysophosphatidyl choline (LPC)-induced demyelinating lesions. The lesions were induced in two areas of the rat brain with either uniform or complex fiber orientations, i.e., in the corpus callosum (cc) and dorsal tegmental tract (dtg), respectively. RAFF4 showed excellent ability to detect demyelinated lesions and good correlation with myelin content in both brain areas. In comparison, diffusion tensor imaging metrices, fractional anisotropy, mean diffusivity and axonal and radial diffusivity, and magnetization transfer (MT) metrices, longitudinal relaxation during off-resonance irradiation and MT ratio, either failed to detect demyelination in dtg or showed lower correlation with myelin density quantified from gold chloride stained histological sections. Good specifity of RAFF4 to myelin was confirmed by its low correlation with cell density assesed from Nissl stained sections as well as its lack of sensitivity to pH changes in the physiological range as tested in heat denaturated bovine serum albumin phantoms. The excellent ability of RAFF4 to detect myelin content and its insensitivity to fiber orientation distribution, gliosis and pH, together with low specific absorption rate, demonstrates the promise of rotating frame of rank n (RAFFn) as a valuable MRI technique for non-invasive imaging of demyelinating lesions.


Social support as a mediator between problem behaviour and gambling: a cross-sectional study among 14-16-year-old Finnish adolescents.
During the adolescent period, risk-taking behaviour increases. These behaviours can compromise the successful transition from adolescence to adulthood. The purpose of this study was to examine social support as a mediator of the relation between problem behaviour and gambling frequency among Finnish adolescents. Data were obtained from the national School Health Promotion Study (SHPS) from the years 2010 and 2011 (N=102 545). Adolescents were classified in the most homogeneous groups based on their problem behaviour via latent class analysis. Path analysis indicated that social support was negatively associated with problem behaviour, and problem behaviour and social support were negatively related (except for social support from friends among boys) to gambling. Social support from parents and school mediated, albeit weakly, the relations between problem behaviour and gambling among girls and boys. Problem behaviour may affect gambling through social support from school and parents. Thus prevention and intervention strategies should focus on strengthening adolescents' social support. In addition, because of the clustering of different problem behaviours instead of concentrating on a single form of problem behaviour multiple-behaviour interventions may have a much greater impact on public health.


An Arabidopsis mutant defective in the general phenylpropanoid pathway.
Mutants of Arabidopsis deficient in a major leaf phenylpropanoid ester, 2-O-sinapoyl-L-malate, were identified by thin-layer chromatographic screening of methanolic leaf extracts from several thousand mutagenized plants. Mutations at a locus designated SIN1 also eliminate accumulation of the sinapic acid esters characteristic of seed tissues. Because of increased transparency to UV light, the sin1 mutants exhibit a characteristic red fluorescence under UV light, whereas wild-type plants have a blue-green appearance due to the fluorescence of sinapoyl malate in the upper epidermis. As determined by in vivo radiotracer feeding experiments, precursor supplementation studies, and enzymatic assays, the defect in the sin1 mutants appears to block the conversion of ferulate to 5-hydroxyferulate in the general phenylpropanoid pathway. As a result, the lignin of the mutant lacks the sinapic acid-derived components typical of wild-type lignin.


Soluble CD93 levels in patients with acute myocardial infarction and its implication on clinical outcome.
Inflammation plays a key role in the pathogenesis of acute myocardial infarction (MI). However, it is unclear whether marker of immune activation will provide prognostic information in these patients. We hypothesized that circulating levels of soluble CD93 (sCD93), a soluble form of transmembrane glycoprotein CD93, is increased in acute MI patients and its level would be associated with clinical outcomes in patients with acute MI. We measured circulating levels of sCD93 in 120 patients with acute MI (63±13 yrs, M∶F = 85∶35) and in 120 age, sex-matched control subjects. In patients with acute MI, clinical characteristics, echocardiographic and laboratory findings were assessed at the time of initial enrollment. The primary outcome was defined as all-cause and cardiovascular death. Circulating sCD93 levels were significantly higher in patients with acute MI than in control subjects (552.1±293.7 vs. 429.8±114.2 ng/mL, p<0.0001). Upon in vitro inflammatory stimulation, increased CD93 shedding was demonstrated in acute MI patients but not in control subjects. During follow up period (median 208 days, 3-1058 days), the primary outcome occurred in 18 (15%) patients (9 cardiovascular deaths). Circulating levels of sCD93 were associated with all cause (p<0.0001) and cardiovascular (p<0.0001) mortality in patients with acute MI. Multivariate Cox regression analysis revealed that initial sCD93 level was found to be an independent predictor of all cause (p = 0.002) and cardiovascular mortality (p = 0.033) when controlled for age and left ventricular ejection fraction. Circulating levels of sCD93 are elevated in patients with acute MI and their levels were associated with adverse clinical outcomes.