A 12-Week Evidence-Based Education Project to Reduce Cardiovascular and Metabolic Risk in Adults With Serious Mental Illness in the Integrated Care Setting.
BACKGROUND: Individuals with serious mental illness (SMI) such as schizophrenia spectrum disorders, severe bipolar disorder, or severe recurrent major depressive disorder have a shorter life expectancy compared with those in the general population. This is largely due to the higher rates of diabetes, hyperlipidemia, and hypertension. Treatment adherence, diet, exercise, and weight management are modifiable risk factors for these cardiometabolic conditions, yet the use of educational lifestyle interventions is not common practice in the clinical setting. AIM: The purpose of this project was to evaluate a 12-week evidence-based diabetes prevention education program integrated into a primary care behavioral health setting for adults with SMI, diabetes, or prediabetes, and who were overweight or obese. METHOD: This project utilized a pretest-posttest intervention design. Outcomes for this project included diabetes knowledge, self-care, and health indicators (blood pressure, hemoglobin A1C, weight, body mass index, and medication adherence). The project also addressed feasibility and acceptability of the program in this setting. RESULTS: Project outcomes showed a reduction in weight, waist circumference, hemoglobin A1C, and blood pressure. Outcomes also indicated an improvement in participants' self-knowledge and self-care. CONCLUSIONS: There was a high attendance rate and overall acceptability and feasibility described by participants. This project highlights the important role that primary care providers can play in providing health education to patients with SMI.


Licence to swill: James Bond's drinking over six decades.
To describe the patterns of alcohol use in James Bond movies over six decades. Film content analysis. Wide range of international locations in 24 James Bond movies (Eon Productions series, 1962-2015). Drinking episodes for Bond and major female characters; alcohol product placement in films; peak estimated blood alcohol concentrations; features relevant to DSM-5 criteria for alcohol use disorder. Bond has drunk heavily and consistently across six decades (109 drinking events; mean, 4.5 events per movie). His peak blood alcohol level was estimated to have been 0.36 g/dL, sufficient to kill some people. We classified him as having severe alcohol use disorder, as he satisfied six of 11 DSM-5 criteria for this condition. Chronic risks for Bond include frequently drinking prior to fights, driving vehicles (including in chases), high stakes gambling, operating complex machinery or devices, contact with dangerous animals, extreme athletic performance, and sex with enemies, sometimes with guns or knives in the bed. Notable trends during the study period included a decline in using alcohol as a weapon (P = 0.023) and an increase in the number of alcohol products in his environment (for alcohol-related product placement: P < 0.001), but his martini consumption has been steady. Drinking by lead female characters and a random selection of 30 of his sexual partners was fairly stable over time, but also occasionally involved binges. James Bond has a severe chronic alcohol problem. He should consider seeking professional help and find other strategies for managing on-the-job stress.


The self in art therapy - Brain-based assessment of the drawing process.
Art therapy plays important role in classical psychological assessment as it allows expressing the subject's sense of self. However, its effectiveness and validity could be impeded by lack of relationship to the patients' neuronal changes in their brain. The aim of our theoretical-empirical paper is to propose a novel brain-based quantitative objective measurement of the self and how it shapes the drawing process. We discuss recent data that how the autocorrelation window (ACW) is related to the temporal continuity of self in current neuroscience and further develop a method to use ACW to measure the temporal continuity of the drawing process, probing it in two case studies. As expected, the schizophrenic subject shows lower ACW values compared to the healthy subject and reflects the well-known deficit in the temporal continuity of the self in schizophrenia. We concluded that ACW and eventually other measures of the brain's spatiotemporal structure might be able to serve as objective markers of the self in the drawing process. As our approach connects brain, self, and drawing process, it provides the theoretical basis for the future development of a brain-based assessment of the self in the drawing process and art therapy.


Achieving integration in mixed methods designs-principles and practices.
Mixed methods research offers powerful tools for investigating complex processes and systems in health and health care. This article describes integration principles and practices at three levels in mixed methods research and provides illustrative examples. Integration at the study design level occurs through three basic mixed method designs-exploratory sequential, explanatory sequential, and convergent-and through four advanced frameworks-multistage, intervention, case study, and participatory. Integration at the methods level occurs through four approaches. In connecting, one database links to the other through sampling. With building, one database informs the data collection approach of the other. When merging, the two databases are brought together for analysis. With embedding, data collection and analysis link at multiple points. Integration at the interpretation and reporting level occurs through narrative, data transformation, and joint display. The fit of integration describes the extent the qualitative and quantitative findings cohere. Understanding these principles and practices of integration can help health services researchers leverage the strengths of mixed methods.


Sex differences in sympathetic neural and limb vascular reactivity to mental stress in humans.
Mental stress elicits a robust and consistent forearm vasodilation, but vascular reactivity in the calf remains inconsistent. It has been reported that calf vascular responses to MS may be sex dependent. Muscle sympathetic nerve activity (MSNA) is an important contributor to calf blood flow (CBF), yet the relations between sex, limb blood flow, and MSNA reactivity to mental stress have not been explored. We hypothesized that mental stress would elicit more dramatic vasodilation of the limbs in women and that this might be explained by reduced MSNA reactivity and/or blunted sympathetic vascular transduction. We measured heart rate (HR), mean arterial pressure (MAP), CBF, calf vascular conductance (CVC), forearm blood flow (FBF), forearm vascular conductance (FVC), and MSNA concurrently in 18 men (age: 23 ± 2 yr) and 16 women (age: 24 ± 2 yr) during 5 min of supine baseline and 5 min of mental stress. Mental stress elicited similar increases in MAP (Δ10 ± 1 vs. Δ11 ± 1 mmHg), HR (Δ16 ± 2 vs. Δ17 ± 2 beats/min), FBF (Δ81 ± 16% vs. Δ83 ± 15%), and FVC (Δ62 ± 13% vs. Δ65 ± 13%) in men and women, respectively. In contrast, CBF (Δ16 ± 8% vs. Δ37 ± 9%, P = 0.036) and CVC (Δ4 ± 7% vs. Δ24 ± 8%, P = 0.036) responses were exaggerated in women compared with men. Changes in FVC were significantly correlated with changes in CVC in women (r = 0.681, P = 0.004) but not in men. MSNA reactivity to mental stress was not different between men and women; however, changes in CVC were negatively correlated with increases of MSNA in men (r = -0.411, P = 0.045) but not in women. In conclusion, our data suggest different patterns of calf vascular reactivity to mental stress in men and women that might relate, in part, to altered vascular transduction of MSNA.


Adverse events associated with unblinded, but not with blinded, statin therapy in the Anglo-Scandinavian Cardiac Outcomes Trial-Lipid-Lowering Arm (ASCOT-LLA): a randomised double-blind placebo-controlled trial and its non-randomised non-blind extension phase.
In blinded randomised controlled trials, statin therapy has been associated with few adverse events (AEs). By contrast, in observational studies, larger increases in many different AEs have been reported than in blinded trials. In the Lipid-Lowering Arm of the Anglo-Scandinavian Cardiac Outcomes Trial, patients aged 40-79 years with hypertension, at least three other cardiovascular risk factors, and fasting total cholesterol concentrations of 6·5 mmol/L or lower, and who were not taking a statin or fibrate, had no history of myocardial infarction, and were not being treated for angina were randomly assigned to atorvastatin 10 mg daily or matching placebo in a randomised double-blind placebo-controlled phase. In a subsequent non-randomised non-blind extension phase (initiated because of early termination of the trial because efficacy of atorvastatin was shown), all patients were offered atorvastatin 10 mg daily open label. We classified AEs using the Medical Dictionary for Regulatory Activities. We blindly adjudicated all reports of four prespecified AEs of interest-muscle-related, erectile dysfunction, sleep disturbance, and cognitive impairment-and analysed all remaining AEs grouped by system organ class. Rates of AEs are given as percentages per annum. The blinded randomised phase was done between February, 1998, and December, 2002; we included 101 80 patients in this analysis (5101 [50%] in the atorvastatin group and 5079 [50%] in the placebo group), with a median follow-up of 3·3 years (IQR 2·7-3·7). The non-blinded non-randomised phase was done between December, 2002, and June, 2005; we included 9899 patients in this analysis (6409 [65%] atorvastatin users and 3490 [35%] non-users), with a median follow-up of 2·3 years (2·2-2·4). During the blinded phase, muscle-related AEs (298 [2·03% per annum] vs 283 [2·00% per annum]; hazard ratio 1·03 [95% CI 0·88-1·21]; p=0·72) and erectile dysfunction (272 [1·86% per annum] vs 302 [2·14% per annum]; 0·88 [0·75-1·04]; p=0·13) were reported at a similar rate by participants randomly assigned to atorvastatin or placebo. The rate of reports of sleep disturbance was significantly lower among participants assigned atorvastatin than assigned placebo (149 [1·00% per annum] vs 210 [1·46% per annum]; 0·69 [0·56-0·85]; p=0·0005). Too few cases of cognitive impairment were reported for a statistically reliable analysis (31 [0·20% per annum] vs 32 [0·22% per annum]; 0·94 [0·57-1·54]; p=0·81). We observed no significant differences in the rates of all other reported AEs, with the exception of an excess of renal and urinary AEs among patients assigned atorvastatin (481 [1·87%] per annum vs 392 [1·51%] per annum; 1·23 [1·08-1·41]; p=0·002). By contrast, during the non-blinded non-randomised phase, muscle-related AEs were reported at a significantly higher rate by participants taking statins than by those who were not (161 [1·26% per annum] vs 124 [1·00% per annum]; 1·41 [1·10-1·79]; p=0·006). We noted no significant differences between statin users and non-users in the rates of other AEs, with the exception of musculoskeletal and connective tissue disorders (992 [8·69% per annum] vs 831 [7·45% per annum]; 1·17 [1·06-1·29]; p=0·001) and blood and lymphatic system disorders (114 [0·88% per annum] vs 80 [0·64% per annum]; 1·40 [1·04-1·88]; p=0·03), which were reported more commonly by statin users than by non-users. These analyses illustrate the so-called nocebo effect, with an excess rate of muscle-related AE reports only when patients and their doctors were aware that statin therapy was being used and not when its use was blinded. These results will help assure both physicians and patients that most AEs associated with statins are not causally related to use of the drug and should help counter the adverse effect on public health of exaggerated claims about statin-related side-effects. Pfizer, Servier Research Group, and Leo Laboratories.


Medusavirus, a Novel Large DNA Virus Discovered from Hot Spring Water.
Recent discoveries of new large DNA viruses reveal high diversity in their morphologies, genetic repertoires, and replication strategies. Here, we report the novel features of medusavirus, a large DNA virus newly isolated from hot spring water in Japan. Medusavirus, with a diameter of 260 nm, shows a T=277 icosahedral capsid with unique spherical-headed spikes on its surface. It has a 381-kb genome encoding 461 putative proteins, 86 of which have their closest homologs in Acanthamoeba, whereas 279 (61%) are orphan genes. The virus lacks the genes encoding DNA topoisomerase II and RNA polymerase, showing that DNA replication takes place in the host nucleus, whereas the progeny virions are assembled in the cytoplasm. Furthermore, the medusavirus genome harbored genes for all five types of histones (H1, H2A, H2B, H3, and H4) and one DNA polymerase, which are phylogenetically placed at the root of the eukaryotic clades. In contrast, the host amoeba encoded many medusavirus homologs, including the major capsid protein. These facts strongly suggested that amoebae are indeed the most promising natural hosts of medusavirus, and that lateral gene transfers have taken place repeatedly and bidirectionally between the virus and its host since the early stage of their coevolution. Medusavirus reflects the traces of direct evolutionary interactions between the virus and eukaryotic hosts, which may be caused by sharing the DNA replication compartment and by evolutionarily long lasting virus-host relationships. Based on its unique morphological characteristics and phylogenomic relationships with other known large DNA viruses, we propose that medusavirus represents a new family, MedusaviridaeIMPORTANCE We have isolated a new nucleocytoplasmic large DNA virus (NCLDV) from hot spring water in Japan, named medusavirus. This new NCLDV is phylogenetically placed at the root of the eukaryotic clades based on the phylogenies of several key genes, including that encoding DNA polymerase, and its genome surprisingly encodes the full set of histone homologs. Furthermore, its laboratory host, Acanthamoeba castellanii, encodes many medusavirus homologs in its genome, including the major capsid protein, suggesting that the amoeba is the genuine natural host from ancient times of this newly described virus and that lateral gene transfers have repeatedly occurred between the virus and amoeba. These results suggest that medusavirus is a unique NCLDV preserving ancient footprints of evolutionary interactions with its hosts, thus providing clues to elucidate the evolution of NCLDVs, eukaryotes, and virus-host interaction. Based on the dissimilarities with other known NCLDVs, we propose that medusavirus represents a new viral family, Medusaviridae.


Development of human brain structural networks through infancy and childhood.
During human brain development through infancy and childhood, microstructural and macrostructural changes take place to reshape the brain's structural networks and better adapt them to sophisticated functional and cognitive requirements. However, structural topological configuration of the human brain during this specific development period is not well understood. In this study, diffusion magnetic resonance image (dMRI) of 25 neonates, 13 toddlers, and 25 preadolescents were acquired to characterize network dynamics at these 3 landmark cross-sectional ages during early childhood. dMRI tractography was used to construct human brain structural networks, and the underlying topological properties were quantified by graph-theory approaches. Modular organization and small-world attributes are evident at birth with several important topological metrics increasing monotonically during development. Most significant increases of regional nodes occur in the posterior cingulate cortex, which plays a pivotal role in the functional default mode network. Positive correlations exist between nodal efficiencies and fractional anisotropy of the white matter traced from these nodes, while correlation slopes vary among the brain regions. These results reveal substantial topological reorganization of human brain structural networks through infancy and childhood, which is likely to be the outcome of both heterogeneous strengthening of the major white matter tracts and pruning of other axonal fibers.


Live-cell observation of cytosolic HIV-1 assembly onset reveals RNA-interacting Gag oligomers.
Assembly of the Gag polyprotein into new viral particles in infected cells is a crucial step in the retroviral replication cycle. Currently, little is known about the onset of assembly in the cytosol. In this paper, we analyzed the cytosolic HIV-1 Gag fraction in real time in live cells using advanced fluctuation imaging methods and thereby provide detailed insights into the complex relationship between cytosolic Gag mobility, stoichiometry, and interactions. We show that Gag diffuses as a monomer on the subsecond timescale with severely reduced mobility. Reduction of mobility is associated with basic residues in its nucleocapsid (NC) domain, whereas capsid (CA) and matrix (MA) domains do not contribute significantly. Strikingly, another diffusive Gag species was observed on the seconds timescale that oligomerized in a concentration-dependent manner. Both NC- and CA-mediated interactions strongly assist this process. Our results reveal potential nucleation steps of cytosolic Gag fractions before membrane-assisted Gag assembly.


How clonal are human mitochondria?
Phylogenetic trees constructed using human mitochondrial sequences contain a large number of homoplasies. These are due either to repeated mutation or to recombination between mitochondrial lineages. We show that a tree constructed using synonymous variation in the protein coding sequences of 29 largely complete human mitochondrial molecules contains 22 homoplasies at 32 phylogenetically informative sites. This level of homoplasy is very unlikely if inheritance is clonal, even if we take into account base composition bias. There must either be 'hypervariable' sites or recombination between mitochondria. We present evidence which suggests that hypervariable sites do not exist in our data. It therefore seems likely that recombination has occurred between mitochondrial lineages in humans.


Dark carbon fixation: an important process in lake sediments.
Close to redox boundaries, dark carbon fixation by chemoautotrophic bacteria may be a large contributor to overall carbon fixation. Still, little is known about the relative importance of this process in lake systems, in spite the potentially high chemoautotrophic potential of lake sediments. We compared rates of dark carbon fixation, bacterial production and oxygen consumption in sediments from four Swedish boreal and seven tropical Brazilian lakes. Rates were highly variable and dark carbon fixation amounted up to 80% of the total heterotrophic bacterial production. The results indicate that non-photosynthetic carbon fixation can represent a substantial contribution to bacterial biomass production, especially in sediments with low organic matter content.


Metformin increases plasma ghrelin in Type 2 diabetes.
* Metformin, unlike the other major antihyperglycaemic drugs, is not associated with weight gain. * Ghrelin is an appetite-stimulating hormone whose concentrations vary in relation to food, obesity and diabetes control. * Reports are conflicting about how metformin affects ghrelin concentrations, and this study was aimed at resolving this issue in patients with Type 2 diabetes. * In this study an increase in ghrelin concentrations was seen in response to metformin treatment in patients with Type 2 diabetes. * This effect was opposite to what might be expected if the effect of metformin on weight control was mediated via suppression of ghrelin. * It is likely that the ghrelin response was secondary to improved glycaemic control. * Meal time changes in appetite and satiety did not correlate with changes in ghrelin, which suggests ghrelin may not be important in meal initiation. Metformin treatment of Type 2 diabetes is not usually associated with weight gain, and may assist with weight reduction. Plasma ghrelin concentrations are inversely associated with obesity and food intake. Metformin might therefore affect ghrelin concentrations, although previous studies have shown variable results in this regard. The primary aim of this study was to determine the effect of metformin on plasma ghrelin, appetite and satiety in patients with Type 2 diabetes. Eighteen patients with Type 2 diabetes were studied before and after 6 weeks of metformin treatment, which was titrated to 1 g b.d. On the study days patients were fed standard meals of 390 kcal at 08.00 and 12.30 h, plasma samples were collected at 15- and 30-min intervals, and appetite and satiety were measured on visual analogue scales. Changes in the area under the concentration-time curves (AUCs) of plasma ghrelin, insulin, glucose, appetite and satiety were assessed and examined for correlations with metformin AUCs. Changes in fasting adiponectin and leptin were also measured. Treatment with metformin increased the mean AUC (07.30-16.30 h) of plasma ghrelin by 24% (P= 0.003), while decreasing those of glucose by 19% (P < 0.001) and insulin by 19% (P= 0.001). No changes were detected in hunger and satiety, or in fasting adiponectin or leptin concentrations. There were no clear correlations between metformin plasma concentrations (AUC) and changes in plasma glucose, insulin or ghrelin. Treatment of Type 2 diabetes with metformin was associated with increased plasma ghrelin concentrations, without associated changes in hunger and satiety.


Visual feedforward control in human locomotion during avoidance of obstacles that change size.
The purpose of the current study was to understand how visual information about an ongoing change in obstacle size is used during obstacle avoidance for both lead and trail limbs. Participants were required to walk in a dark room and to step over an obstacle edged with a special tape visible in the dark. The obstacle's dimensions were manipulated one step before obstacle clearance by increasing or decreasing its size. Two increasing and two decreasing obstacle conditions were combined with seven control static conditions. Results showed that information about the obstacle's size was acquired and used to modulate trail limb trajectory, but had no effect on lead limb trajectory. The adaptive step was influenced by the time available to acquire and process visual information. In conclusion, visual information about obstacle size acquired during lead limb crossing was used in a feedforward manner to modulate trail limb trajectory.


Repair in rheumatoid arthritis, current status. Report of a workshop at OMERACT 8.
Repair of structural damage in rheumatoid arthritis has drawn much attention with newly available effective treatments. A workshop was held at OMERACT 8 to update current knowledge on the validity of the concept of repair and on the assessment of repair. In preparation for the workshop several studies were performed and the results were presented. This was followed by a discussion and voting on statements on various aspects of repair. A majority of participants agreed that results of the new studies strengthen the validity of the concept of repair, and that repair can be assessed on radiographs. There was less agreement on the best means of measurement and there was a plea for more extensive reporting of data, i.e., not limited to sum scores of all joints together. The conclusions of the workshop mean a big step forward in the acceptance and assessment of repair.


Time in Range in Pregnancy: Is There a Role?
Continuous glucose monitoring (CGM) is being used with increasing frequency as an adjunct to self-monitoring of blood glucose in pregnancy, and novel targets based on CGM data are becoming standardized. This adoption of CGM is the result of its improving accuracy, patient preference, and evolving data demonstrating associations of novel targets such as time in range (TIR) with pregnancy and neonatal outcomes. A greater understanding of the relationship of various CGM metrics to outcomes in pregnancy complicated by diabetes is needed. It is clear that TIR parameters need to be uniquely lower for pregnant women than for nonpregnant individuals. CGM technology is also an integral part of hybrid closed-loop insulin delivery systems. These insulin delivery systems will be a significant advance in the management of diabetes during pregnancy if they can achieve the pre- and postprandial targets required for pregnancy and optimize TIR.


Sinking particles promote vertical connectivity in the ocean microbiome.
The sinking of organic particles formed in the photic layer is a main vector of carbon export into the deep ocean. Although sinking particles are heavily colonized by microbes, so far it has not been explored whether this process plays a role in transferring prokaryotic diversity from surface to deep oceanic layers. Using Illumina sequencing of the 16S rRNA gene, we explore here the vertical connectivity of the ocean microbiome by characterizing marine prokaryotic communities associated with five different size fractions and examining their compositional variability from surface down to 4,000 m across eight stations sampled in the Atlantic, Pacific, and Indian Oceans during the Malaspina 2010 Expedition. Our results show that the most abundant prokaryotes in the deep ocean are also present in surface waters. This vertical community connectivity seems to occur predominantly through the largest particles because communities in the largest size fractions showed the highest taxonomic similarity throughout the water column, whereas free-living communities were more isolated vertically. Our results further suggest that particle colonization processes occurring in surface waters determine to some extent the composition and biogeography of bathypelagic communities. Overall, we postulate that sinking particles function as vectors that inoculate viable particle-attached surface microbes into the deep-sea realm, determining to a considerable extent the structure, functioning, and biogeography of deep ocean communities.


Increased hepatitis C virus vaccine clinical trial literacy following a brief intervention among people who inject drugs.
While people who inject drugs are at high risk of hepatitis C virus (HCV) infection and will be the target population for future HCV vaccine trials, little is known about clinical trial literacy (CTL) in this group. We assessed the impact of a brief intervention (BI) designed to improve HCV vaccine CTL among people who inject drugs in Sydney, Australia. People who inject drugs enrolled in a community-based prospective observational study between November 2008 and September 2010 (n = 102) completed a CTL assessment followed immediately by the BI. Post-test assessment was conducted at 24 weeks. The median age of the sample was 27 years, 73% were male and 60% had 10 or less years of schooling. The median time since first injection was 5 years and 20% reported daily or more frequent injecting. The mean number of correct responses increased from 5.3 to 6.3/10 (t = -4.2; 101df, P < 0.001) 24 weeks post-intervention. Statistically significant differences were observed for three knowledge items with higher proportions of participants correctly answering questions related to randomisation (P = 0.002), blinding (P = 0.005) and vaccine-induced seropositivity (P = 0.003) post-intervention. A significant increase in HCV vaccine CTL was observed, suggesting that new and relatively novel concepts can be learned and recalled in this group. These findings support the feasibility of future trials among this population. [Correction added on 21 November 2012, after first online publication: T-score for mean number of correct responses was corrected to '-4.2' in the Results section.]


Multimodal medical case retrieval using the Dezert-Smarandache theory.
Most medical images are now digitized and stored with semantic information, leading to medical case databases. They may be used for aid to diagnosis, by retrieving similar cases to those in examination. But the information are often incomplete, uncertain and sometimes conflicting, so difficult to use. In this paper, we present a Case Based Reasoning (CBR) system for medical case retrieval, derived from the Dezert-Smarandache theory, which is well suited to handle those problems. We introduce a case retrieval specific frame of discernment theta, which associates each element of theta with a case in the database; we take advantage of the flexibility offered by the DSmT's hybrid models to finely model the database. The system is designed so that heterogeneous sources of information can be integrated in the system: in particular images, indexed by their digital content, and symbolic information. The method is evaluated on two classified databases: one for diabetic retinopathy follow-up (DRD) and one for screening mammography (DDSM). On these databases, results are promising: the retrieval precision at five reaches 81.8% on DRD and 84.8% on DDSM.


Pregabalin as an adjunct to phenobarbital, potassium bromide, or a combination of phenobarbital and potassium bromide for treatment of dogs with suspected idiopathic epilepsy.
To assess tolerability and short-term efficacy of oral administration of pregabalin as an adjunct to phenobarbital, potassium bromide, or a combination of phenobarbital and potassium bromide for treatment of dogs with poorly controlled suspected idiopathic epilepsy. Open-label, noncomparative clinical trial. 11 client-owned dogs suspected of having idiopathic epilepsy that was inadequately controlled with phenobarbital, potassium bromide, or a combination of these 2 drugs. Dogs were treated with pregabalin (3 to 4 mg/kg [1.4 to 1.8 mg/lb], PO, q 8 h) for 3 months. Number of generalized seizures in the 3 months before and after initiation of pregabalin treatment was recorded. Number of responders (>or= 50% reduction in seizure frequency) was recorded, and seizure frequency before and after initiation of pregabalin treatment was compared by use of a nonparametric Wilcoxon signed rank test. Seizures were significantly reduced (mean, 57%; median, 50%) after pregabalin administration in the 9 dogs that completed the study; 7 were considered responders with mean and median seizure reductions of 64% and 58%, respectively. Adverse effects for pregabalin were reported in 10 dogs. Mean and median plasma pregabalin concentrations for all dogs were 6.4 and 7.3 microg/mL, respectively. Pregabalin may hold promise as a safe and effective adjunct anticonvulsant drug for epileptic dogs poorly controlled with the standard drugs phenobarbital or potassium bromide. Adverse effects of pregabalin appeared to be mild. Additional studies with larger numbers of dogs and longer follow-up intervals are warranted.


Effects of calcium on the kinetics of a model disjunctive ligand exchange reaction: implications for dynamic trace metal ion speciation.
Trace metal ion speciation in natural waters is often under kinetic control due to slow exchange reactions involving multidentate ligands (both natural and anthropogenic) and constituent ions (e.g. calcium). Incomplete understanding of the kinetic behavior (rates, rate laws, and mechanisms) of multidentate ligand exchange reactions hinders prediction of metal ion bioavailability and mobility in these systems. Here, we aim to improve understanding (1) by examining the mechanism by which calcium suppresses ligand exchange rates and (2) by developing conceptual tools for predicting the kinetic behavior in environmental systems. Using capillary electrophoresis, we examined the effect of calcium concentration on the kinetics of a model disjunctive multidentate ligand exchange reaction between nickel(ii)-nitrilotriacetate (NiNTA) and trans-1,2-cyclohexylenedinitrilotetraacetate (CDTA) at constant temperature (25 °C) and ionic strength (10 mM). Initial NiCDTA formation rates under varying reactant and calcium concentrations were fit with a single disjunctive ligand exchange kinetic model. While the overall reaction pathway is disjunctive under all conditions studied, the presence of calcium causes a non-linear decrease in reaction rates and alters the overall reaction order with respect to free multidentate ligand concentration. The overall rate law is affected by the relative calcium affinity of exchanging ligands, which controls the balance of product formation and reactant reformation rates from the free metal ion intermediate. The log ratio of these rates is proposed as a metric for determining the appropriate simplified form of the rate law for disjunctive ligand exchange reactions. The implications for in situ ligand lability assays and metal ion bioavailability are discussed.


Delivery of iron to human cells by bovine transferrin. Implications for the growth of human cells in vitro.
Following suggestions that transferrin present in fetal-bovine serum, a common supplement used in tissue-culture media, may not bind well to human cells, we have isolated the protein and investigated its interaction with both human and bovine cells. Bovine transferrin bound to a human cell line, K562, at 4 degrees C with a kd of 590 nM, whereas human transferrin bound with a kd of 3.57 nM, a 165-fold difference. With a bovine cell line, NBL4, bovine transferrin bound with the higher affinity, kd 9.09 nM, whereas human transferrin bound with a kd of 41.7 nM, only a 5-fold difference. These values were reflected in an 8.6-fold difference in the rate of iron delivery by the two proteins to human cells, whereas delivery to bovine cells was the same. Nevertheless, the bovine transferrin was taken up by the human cells by a specific receptor-mediated process. Human cells cultured in bovine diferric transferrin at 40 micrograms/ml, the concentration expected in the presence of 10% fetal-bovine serum, failed to thrive, whereas cells cultured in the presence of human transferrin proliferated normally. These results suggest that growth of human cells in bovine serum could give rise to a cellular iron deficiency, which may in turn lead to the selection of clones of cells adapted for survival with less iron. This has important consequences for the use of such cells as models, since they may have aberrant iron-dependent pathways and perhaps other unknown alterations in cell function.


Tree cover in sub-Saharan Africa: rainfall and fire constrain forest and savanna as alternative stable states.
Savannas are known as ecosystems with tree cover below climate-defined equilibrium values. However, a predictive framework for understanding constraints on tree cover is lacking. We present (a) a spatially extensive analysis of tree cover and fire distribution in sub-Saharan Africa, and (b) a model, based on empirical results, demonstrating that savanna and forest may be alternative stable states in parts of Africa, with implications for understanding savanna distributions. Tree cover does not increase continuously with rainfall, but rather is constrained to low (<50%, "savanna") or high tree cover (>75%, "forest"). Intermediate tree cover rarely occurs. Fire, which prevents trees from establishing, differentiates high and low tree cover, especially in areas with rainfall between 1000 mm and 2000 mm. Fire is less important at low rainfall (<1000 mm), where rainfall limits tree cover, and at high rainfall (>2000 mm), where fire is rare. This pattern suggests that complex interactions between climate and disturbance produce emergent alternative states in tree cover. The relationship between tree cover and fire was incorporated into a dynamic model including grass, savanna tree saplings, and savanna trees. Only recruitment from sapling to adult tree varied depending on the amount of grass in the system. Based on our empirical analysis and previous work, fires spread only at tree cover of 40% or less, producing a sigmoidal fire probability distribution as a function of grass cover and therefore a sigmoidal sapling to tree recruitment function. This model demonstrates that, given relatively conservative and empirically supported assumptions about the establishment of trees in savannas, alternative stable states for the same set of environmental conditions (i.e., model parameters) are possible via a fire feedback mechanism. Integrating alternative stable state dynamics into models of biome distributions could improve our ability to predict changes in biome distributions and in carbon storage under climate and global change scenarios.


Breakfast replacement with a low-glycaemic response liquid formula in patients with type 2 diabetes: a randomised clinical trial.
Low-glycaemic index diets reduce glycated Hb (HbA1c) in patients with type 2 diabetes, but require intensive dietary support. Using a liquid meal replacement with a low glycaemic response (GR) may be an alternative dietary approach. In the present study, we investigated whether breakfast replacement with a low-GR liquid meal would reduce postprandial glycaemia and/or improve long-term glycaemia. In the present randomised, controlled, cross-over design, twenty patients with type 2 diabetes consumed either a breakfast replacement consisting of an isoenergetic amount of Glucerna SR or a free-choice breakfast for 3 months. Postprandial AUC levels were measured using continuous glucose measurement at home. After the 3-month dietary period, meal profiles and oral glucose tolerance were assessed in the clinical setting. The low-GR liquid meal replacement reduced the AUC of postprandial glucose excursions at home compared with a free-choice control breakfast (estimated marginal mean 141 (95 % CI 114, 174) v. estimated marginal mean 259 (95 % CI 211, 318) mmol × min/l; P= 0·0002). The low-GR liquid meal replacement also reduced glucose AUC levels in the clinical setting compared with an isoenergetic control breakfast (low GR: median 97 (interquartile range (IQR) 60-188) mmol × min/l; control: median 253 (IQR 162-386) mmol × min/l; P< 0·001). However, the 3-month low-GR liquid meal replacement did not affect fasting plasma glucose, HbA1c or lipid levels, and even slightly reduced oral glucose tolerance. In conclusion, the low-GR liquid meal replacement is a potential dietary approach to reduce postprandial glycaemia in patients with type 2 diabetes. However, clinical trials into the effects of replacing multiple meals on long-term glycaemia in poorly controlled patients are required before a low-GR liquid meal replacement can be adopted as a dietary approach to the treatment of type 2 diabetes.


Genetic determinants of telomere length and risk of common cancers: a Mendelian randomization study.
Epidemiological studies have reported inconsistent associations between telomere length (TL) and risk for various cancers. These inconsistencies are likely attributable, in part, to biases that arise due to post-diagnostic and post-treatment TL measurement. To avoid such biases, we used a Mendelian randomization approach and estimated associations between nine TL-associated SNPs and risk for five common cancer types (breast, lung, colorectal, ovarian and prostate cancer, including subtypes) using data on 51 725 cases and 62 035 controls. We then used an inverse-variance weighted average of the SNP-specific associations to estimate the association between a genetic score representing long TL and cancer risk. The long TL genetic score was significantly associated with increased risk of lung adenocarcinoma (P = 6.3 × 10(-15)), even after exclusion of a SNP residing in a known lung cancer susceptibility region (TERT-CLPTM1L) P = 6.6 × 10(-6)). Under Mendelian randomization assumptions, the association estimate [odds ratio (OR) = 2.78] is interpreted as the OR for lung adenocarcinoma corresponding to a 1000 bp increase in TL. The weighted TL SNP score was not associated with other cancer types or subtypes. Our finding that genetic determinants of long TL increase lung adenocarcinoma risk avoids issues with reverse causality and residual confounding that arise in observational studies of TL and disease risk. Under Mendelian randomization assumptions, our finding suggests that longer TL increases lung adenocarcinoma risk. However, caution regarding this causal interpretation is warranted in light of the potential issue of pleiotropy, and a more general interpretation is that SNPs influencing telomere biology are also implicated in lung adenocarcinoma risk.


Vascular reconstruction in limb salvage operations for soft tissue tumors of the extremities.
Eight patients with vascular threatening soft tissue tumors underwent wide excision of the tumors followed by vascular reconstruction. Superficial femoral vessels were resected in 6 patients and popliteal vessels in 2 patients. Arterial reconstruction was performed with a synthetic graft in 4 and a reversed saphenous vein graft in 4. Venous reconstruction was performed with a synthetic graft in 5 and a saphenous vein graft in 2. Adjuvant multimodality treatment was used in 6 patients. After an average of 30 months, the revascularized vessels were found to be patent in 5 patients with arterial reconstruction and in 1 patient with venous reconstruction. Six patients maintained functional extremities. Two other patients had to have amputations, 1 because of a deep infection that involved the grafts and the other because of an acute occlusion of the reconstructed artery. Edema of the leg, which was treated successfully with an elastic support and elevation of the leg, has been observed in all patients after operation. No local recurrence has been observed with an average followup period of 42.5 months. Pulmonary metastasis developed in 2 patients, and 1 of them underwent a pulmonary metastasectomy. Limb salvage in vascular threatening soft tissue tumors using vascular reconstruction techniques is useful and preferable in some patients.


Maternal bladder cancer diagnosed at routine first-trimester obstetric ultrasound examination.
Bladder cancer is exceedingly rare in pregnancy and most commonly presents with gross hematuria. We describe two patients with the incidental finding of maternal bladder masses identified during routine first-trimester obstetric ultrasonographic evaluation and an ultimate diagnosis of carcinoma. After referral for urology evaluation and biopsy confirmation of bladder cancer, patients underwent surgical resection during their pregnancies without the need for further treatment and had uncomplicated pregnancy courses. The distended maternal urinary bladder at the time of first-trimester ultrasonographic evaluation offers a unique opportunity for examination and early diagnosis of incidental maternal bladder carcinoma.


Individuals with occupational allergy to detergent enzymes display a differential transcriptional regulation and cellular immune response.
In spite of significant safety measures, allergy to industrial enzymes remains a major concern. The increasing prevalence of occupational allergy emphasizes the need to investigate the functional properties of enzyme-exposed dendritic cells (DCs), as DCs possess a potent ability to activate allergen-specific T cells. This study aims at elucidating the molecular mechanisms underlying allergic immune responses to lipase, an industrial enzyme. For this purpose, we studied the effect of both hypoallergenic and wild-type lipase on the transcriptional regulation in DCs and their stimulatory effect on memory CD4+ T cells. Five individuals with documented lipase allergy were tested for specific serum IgE. DCs from these individuals, stimulated with lipases, were assayed for their ability to affect proliferation and polarization of memory T cells. The effect of lipases on transcriptional activity in DCs was evaluated using global expression analysis. Lipase-specific IgE levels varied considerably between donors, with donor 4 exhibiting highest levels, and a potent specific CD4+ T cell recall response was demonstrated only for donor 4. No difference was detected in cytokine profile when T cells from donor 4 were co-cultured with DCs pulsed with either hypoallergenic or wild-type lipase, as demonstrated by high IL-4 and IL-13, and low IFN-gamma production. However, the lipases induced different genetic signatures in DCs from donor 4, as compared with the non-responders. DCs from individuals with clinically diagnosed allergy to lipase displayed a differential response to stimulation with hypoallergenic and wild-type lipase in vitro. Only allergen-pulsed DCs from donor 4 were able to induce CD4+ T cell proliferation. The lipase-specific T cells displayed a T-helper type 2 phenotype, which was not altered by hypoallergenic lipase-pulsed DCs. Furthermore, DCs derived from donor 4 and stimulated with either of the lipases displayed different transcriptional profiles, as compared with the other donors. These signatures represent genes of potential importance for an immunoregulatory role of DC in an ongoing allergic response.


Assessment of the natural history of forefoot bursae using ultrasonography in patients with rheumatoid arthritis: a twelve-month investigation.
To determine the natural history and clinical significance of forefoot bursae over a 12-month period in patients with rheumatoid arthritis (RA). Patients with RA (n=149) attending rheumatology outpatient clinics were assessed at baseline. A total of 120 participants, mean±SD age 60.7±12.1 years and mean±SD disease duration 12.99±10.4 years, completed the 12-month followup (98 women, 22 men, 93 rheumatoid factor positive, 24 rheumatoid factor negative, and 3 unknown). Musculoskeletal ultrasound (US) was used to identify forefoot bursae in all of the participants. Clinical markers of disease activity (well-being visual analog scale [VAS], erythrocyte sedimentation rate [ESR], C-reactive protein [CRP] level, and Disease Activity Score in 28 joints [DAS28]) and foot symptoms on the Leeds Foot Impact Scale (LFIS) Questionnaire were recorded on both occasions. Presence of US-detectable forefoot bursae was identified in 93.3% of returnee (n=120) participants (individual mean 3.7, range 0-11) at baseline. Significant associations were identified between bursae presence and patient-reported foot impact for impairment/footwear (LFISIF ; baseline: r=0.226, P=0.013 and 12 months: r=0.236, P=0.009) and activity limitation/participation restriction (LFISAP; baseline: r=0.254, P=0.005 and 12 months: r=0.235, P=0.010). After 12 months, 42.5% of participants had an increase in the number of US-detectable forefoot bursae and 45% of participants had a decrease. Changes in bursae number significantly correlated with changes in LFISIF (r=0.216, P=0.018) and LFISAP (r=0.193, P=0.036). No significant associations were identified between changes in bursae and changes in global well-being VAS, ESR, CRP level, or DAS28. The findings of this study suggest that forefoot bursae may regress or hypertrophy over time in patients with RA, and that these changes may be associated with self-reported foot impairment and activity restriction.


Common variants near ATM are associated with glycemic response to metformin in type 2 diabetes.
Metformin is the most commonly used pharmacological therapy for type 2 diabetes. We report a genome-wide association study for glycemic response to metformin in 1,024 Scottish individuals with type 2 diabetes with replication in two cohorts including 1,783 Scottish individuals and 1,113 individuals from the UK Prospective Diabetes Study. In a combined meta-analysis, we identified a SNP, rs11212617, associated with treatment success (n = 3,920, P = 2.9 × 10(-9), odds ratio = 1.35, 95% CI 1.22-1.49) at a locus containing ATM, the ataxia telangiectasia mutated gene. In a rat hepatoma cell line, inhibition of ATM with KU-55933 attenuated the phosphorylation and activation of AMP-activated protein kinase in response to metformin. We conclude that ATM, a gene known to be involved in DNA repair and cell cycle control, plays a role in the effect of metformin upstream of AMP-activated protein kinase, and variation in this gene alters glycemic response to metformin.


Diagnostic Imaging: Appropriate and Safe Use.
The use of diagnostic radiography has doubled in the past two decades. Image Gently (children) and Image Wisely (adults) are multidisciplinary initiatives that seek to reduce radiation exposure by eliminating unnecessary procedures and offering best practices. Patients with an estimated glomerular filtration rate less than 30 mL per minute per 1.73 m2 may have increased risk of nephropathy when exposed to iodinated contrast media and increased risk of nephrogenic systemic fibrosis when exposed to gadolinium-based contrast agents. American College of Radiology Appropriateness Criteria can help guide specific diagnostic imaging choices. Noncontrast head computed tomography is the first-line modality when a stroke is suspected. Magnetic resonance imaging stroke protocols and computed tomography perfusion scans can augment evaluation and potentially expand pharmacologic and endovascular therapy timeframes. Imaging should be avoided in patients with uncomplicated headache syndromes unless the history or physical examination reveals red flag features. Cardiac computed tomography angiography, stress echocardiography, and myocardial perfusion scintigraphy (nuclear stress test) are appropriate for patients with chest pain and low to intermediate cardiovascular risk and have comparable sensitivity and specificity. Computed tomography pulmonary angiography is the preferred test for high-risk patients or those with a positive d-dimer test result, and ventilation-perfusion scintigraphy is reserved for patients with an estimated glomerular filtration rate less than 30 mL per minute per 1.73 m2 or a known contrast allergy. Computed tomography with intravenous contrast is preferred for evaluating adults with suspected appendicitis; however, ultrasonography should precede computed tomography in children, and definitive treatment should be initiated if positive. Ultrasonography is the first-line modality for assessing right upper quadrant pain suggestive of biliary disease. Mass size and patient age dictate surveillance recommendations for adnexal masses. Imaging should not be performed for acute (less than six weeks) low back pain unless red flag features are found on patient history. Ultrasonography should be used for the evaluation of suspicious thyroid nodules identified incidentally on computed tomography.


Overcoming public health risks to staff during the management of waste from healthcare facilities.
Purpose The management of public health risks is a key focus for the European Union. One of the key factors that has been shown to pose a public health risk is that of the management of needles from healthcare facilities. The paper aims to discuss this issue. Design/methodology/approach Using audits of two case study hospitals based in northern Italy, this study sought to examine the key factors that resulted in needle stick injuries amongst staff and suggest measures to minimise these risks. Findings The number of needle stick injuries was influenced by various key factors including the time period during the year, the length of time employed, the location within the site, staff category and working hours. Practical implications Suggestions for overcoming the risk factors, including redesigning working patterns, staff training and awareness building, and the use of safety-engineered devices are outlined. Originality/value This study examined the incidence of needle stick injuries amongst healthcare staff in two Italian hospitals, which was lower than in other countries due to various factors, including recapping of needles not being allowed, the introduction of self-retractable needles and awareness campaigns about the correct disposal procedures of potentially infectious waste.


Utility of Gastric and Duodenal Biopsies During Follow-up Endoscopy in Children With Eosinophilic Esophagitis.
Eosinophilic esophagitis (EoE) is a chronic antigen-mediated immune disorder of the esophagus. Consensus guidelines recommend obtaining esophageal, gastric, and duodenal biopsies at diagnostic endoscopy when EoE is suspected. The utility of repeated gastric and duodenal biopsies during follow-up endoscopy in patients previously diagnosed with EoE is not established. The aim of the present study was to explore the role of gastric and duodenal biopsies in children with an established diagnosis of EoE undergoing repeat endoscopy to assess histological response to treatment. Retrospective chart review of children diagnosed with EoE at a tertiary care center was conducted. A total of 160 patients with EoE with demographic clinical, endoscopic, and histological data at diagnosis and follow-up endoscopy were included. The frequency of gastric and duodenal biopsies at follow-up endoscopy with abnormal histology and their correlation to endoscopic findings was determined. At follow-up endoscopy, 83% (132/160) of patients had gastric and 74% (118/160) had duodenal biopsies. Histology was normal in 81% of gastric and 92% of duodenal biopsies. The most frequent gastric abnormalities were chemical and inactive chronic gastritis. The most frequent duodenal abnormality was villous blunting with increased intraepithelial lymphocytes. Two patients with normal gastric and duodenal histology progressed to eosinophilic gastroenteritis at follow-up endoscopy. Gastric and duodenal biopsies obtained in EoE patients during follow-up endoscopy show pathology in a minority of patients, increase costs, and may add potential risk of adverse events. Large multicenter, prospective studies of endoscopic practice during follow-up of EoE are warranted to provide evidence supporting best practices.


Natural selection at genomic regions associated with obesity and type-2 diabetes: East Asians and sub-Saharan Africans exhibit high levels of differentiation at type-2 diabetes regions.
Different populations suffer from different rates of obesity and type-2 diabetes (T2D). Little is known about the genetic or adaptive component, if any, that underlies these differences. Given the cultural, geographic, and dietary variation that accumulated among humans over the last 60,000 years, we examined whether loci identified by genome-wide association studies for these traits have been subject to recent selection pressures. Using genome-wide SNP data on 938 individuals in 53 populations from the Human Genome Diversity Panel, we compare population differentiation and haplotype patterns at these loci to the rest of the genome. Using an "expanding window" approach (100-1,600 kb) for the individual loci as well as the loci as ensembles, we find a high degree of differentiation for the ensemble of T2D loci. This differentiation is most pronounced for East Asians and sub-Saharan Africans, suggesting that these groups experienced natural selection at loci associated with T2D. Haplotype analysis suggests an excess of obesity loci with evidence of recent positive selection among South Asians and Europeans, compared to sub-Saharan Africans and Native Americans. We also identify individual loci that may have been subjected to natural selection, such as the T2D locus, HHEX, which displays both elevated differentiation and extended haplotype homozygosity in comparisons of East Asians with other groups. Our findings suggest that there is an evolutionary genetic basis for population differences in these traits, and we have identified potential group-specific genetic risk factors.


Treatment of streptococcal pharyngitis with amoxycillin once a day.
To evaluate treatment of group A beta haemolytic streptococcal pharyngitis with amoxycillin once daily compared with phenoxymethylpenicillin three or four times a day. Randomised controlled study of consecutive patients presenting with symptoms suggestive of group A beta haemolytic streptococcal pharyngitis in whom culture of a throat swab yielded positive results. Five family medicine practices. 157 patients aged over 3 years who required treatment with antibiotics. Clinical response, bacteriological response, days at work and school lost, and compliance. During the period of the study 393 patients presented with symptoms suggesting streptococcal pharyngitis; 157 of them had throat swabs that yielded positive results on culture. Eighty two were treated with phenoxymethylpenicillin and 75 with amoxycillin. No difference was observed in the clinical response, days at work and school lost (139 days for 64 patients taking phenoxymethylpenicillin v 100 days for 57 patients taking amoxycillin; p > 0.2), or residual positive cultures after two days (6 (7.3%) v 3 (4%); p > 0.5). A significant difference in the bacteriological response was found after 14 days (5 (6.1%) v 0; p < 0.04) with no positive cultures observed in the amoxycillin group. These findings support the hypothesis that amoxycillin once daily is as effective as phenoxymethylpenicillin in the treatment of group A beta haemolytic streptococcal pharyngitis.


Phenotype in retinol deficiency due to a hereditary defect in retinol binding protein synthesis.
To describe the phenotype caused by a retinol deficiency in a family with compound heterozygous missense mutations (Ile41Asn and Gly75Asp) in the gene for serum retinol binding protein (RBP). The two affected sisters, 17 (BR) and 13 (MR) years old, were examined clinically and with perimetry, color vision tests, dark adaptometry, rod- and cone-isolated electroretinograms (ERGs), multifocal ERGs, electrooculograms (EOGs), and laboratory tests. There were no complaints besides night vision problems and no history of systemic disease. Visual acuity was reduced to 20/40 (BR) and 20/25 (MR). Anterior segments were normal except for a discrete iris coloboma. Both patients showed a typical "fundus xerophthalmicus," featuring a progressed atrophy of the retinal pigment epithelium. Dark adaptation thresholds were elevated. In the scotopic ERG, only reduced mixed responses were recordable. The photopic ERG was reduced in BR and normal in MR; implicit times were highly (BR) to slightly (MR) elevated. There was no (BR) to little (MR) light reaction in the EOG. All-trans retinol levels were 0.19 microM and 0.18 microM (normal range, 0.7-1.5 microM) for BR and MR, respectively, and did not increase in a dose-response test. RBP was below detection threshold, and retinyl esters were normal. Both affected siblings had no detectable serum RBP, one sixth of normal retinol levels, and normal retinyl esters. The retinal pigment epithelium was severely affected, but besides acne there were no changes to other organs. This gives evidence for an alternative tissue source of vitamin A, presumably retinyl esters from chylomicron remnants. The normal retinol levels in the tear fluid explain the lack of xerophthalmia. However, considering the role of RBP in the tear fluid and, during development, in the yolk sac there is also evidence that there are organ-specific RBP forms not affected by the genetic defect.


Bifocal cortical electrical stimulation for pain by interdural implantation of the electrodes.
Chronic electrical cortical stimulation (ECS) is an evolving therapy for alleviating treatment-refractory chronic pain syndromes. In this report, the authors describe a modified technique of ECS that involves resection of dural strips and interdural placement of the electrodes as a patch, and bifocal stimulation by implanting 2 electrode strips, that is, one over the motor and one over the sensory cortices. The technique was used in 4 patients with treatment-refractory pain syndromes: a 76-year-old woman with poststroke central pain, 2 women, (71 and 73 years old) with trigeminal pain, and a 44-year-old man with phantom limb pain. All 4 patients experienced a sustained significant improvement in the intensity of pain and have gained a substantially improved functionality and quality of life. An important finding in these patients was the constancy of impedance within a narrow values range throughout the postoperative period. For the cases, the follow-up exceeds 24, 15, 12, and 9 months. The factors affecting the efficacy of ECS are discussed. In the authors' opinion, interdural implantation of the electrodes holds the promise to improve the efficacy and consistency of ECS compared with the standard epidural or subdural implantation without increasing the risk of the procedure. The technical considerations and the potential therapeutic advantages of the interdural bifocal approach are discussed.


Development of Human-Like scFv-Fc Neutralizing Botulinum Neurotoxin E.
Botulinum neurotoxins (BoNTs) are considered to be the most toxic substances known on earth and are responsible for human botulism, a life-threatening disease characterized by flaccid muscle paralysis that occurs naturally by food-poisoning or colonization of the gastrointestinal tract by BoNT-producing clostridia. BoNTs have been classified as category A agent by the Centers of Disease Control and Prevention (CDC) and are listed among the six agents with the highest risk to be used as bioweapons. Neutralizing antibodies are required for the development of effective anti-botulism therapies to deal with the potential risk of exposure. In this study, a macaque (Macaca fascicularis) was immunized with recombinant light chain of BoNT/E3 and an immune phage display library was constructed. After a multi-step panning, several antibody fragments (scFv, single chain fragment variable) with nanomolar affinities were isolated, that inhibited the endopeptidase activity of pure BoNT/E3 in vitro by targeting its light chain. Furthermore, three scFv were confirmed to neutralize BoNT/E3 induced paralysis in an ex vivo mouse phrenic nerve-hemidiaphragm assay. The most effective neutralization (20LD50/mL, BoNT/E3) was observed with scFv ELC18, with a minimum neutralizing concentration at 0.3 nM. Furthermore, ELC18 was highly effective in vivo when administered as an scFv-Fc construct. Complete protection of 1LD50 BoNT/E3 was observed with 1.6 ng/dose in the mouse flaccid paralysis assay. These scFv-Fcs antibodies are the first recombinant antibodies neutralizing BoNT/E by targeting its light chain. The human-like nature of the isolated antibodies is predicting a good tolerance for further clinical development.


Naphthazarin and methylnaphthazarin cause vascular dysfunction by impairment of endothelium-derived nitric oxide and increased superoxide anion generation.
The effects of the naphthoquinone analogue, naphthazarin (Nap), and its derivative, methylnaphthazarin (MetNap), on vascular reactivity were studied using isolated rat aortic rings and human umbilical vein endothelial cells (HUVECs). In this study, we determined vessel tension, nitric oxide (NO) formation, endothelial nitric oxide synthase (eNOS) activity, eNOS protein expression, and superoxide anion (O2*-) generation in an effort to evaluate the effect of Nap and MetNap on the impairment of the NO-mediated pathway. Lower concentrations of Nap (0.01-1 microM) and MetNap (1-10 microM) concentration-dependently enhanced phenylephrine (PE)-induced vasocontraction and abrogated acetylcholine (ACh)-induced vasorelaxation in an endothelium-dependent manner. On HUVECs, both Nap and MetNap concentration-dependently inhibited NO formation induced by A23187, and also partially inhibited nitric oxide synthase (NOS) activity. eNOS protein expression by HUVECs was not affected by treatment with Nap or MetNap, even within 24h. These data suggest that Nap and MetNap might act as inhibitors of nitric oxide synthesis in the endothelium. In addition, Nap and MetNap were also shown to generate O2*- on HUVECs with short-term treatment. We concluded that Nap and MetNap inhibited agonist-induced relaxation and induced vasocontraction in an endothelium-dependent manner, and these effects might have been due to modification of the NO content by inhibition of NOS activity and bioinactivation through O2*- generation.


Epidermis-specific gene expression in Pachyphytum.
Transcripts of exclusively epidermis-specific expression were obtained by differential screening of a cDNA library prepared from isolated epidermis tissue of a succulent plant, Pachyphytum. Six transcripts were selected and characterized by RNA gel blot hybridization. Five cDNAs represented abundant mRNAs found exclusively in the epidermis. In situ hybridizations with three of these transcripts further substantiated their epidermal location. While two transcripts were present in all cells of the epidermis, one transcript was enriched in subsidiary cells of the stomatal complexes. All of the transcripts were highly expressed in the protoderm of the shoot apical meristem. DNA sequence determination indicated that the mRNAs identified represent novel sequences, encoding yet unknown plant functions. These transcripts, their control elements, and their encoded functions should help to advance our understanding of epidermal cell determination and differentiation.


Mutant PIK3CA Induces EMT in a Cell Type Specific Manner.
Breast cancer is characterized into different molecular subtypes, and each subtype is characterized by differential gene expression that are associated with distinct survival outcomes in patients. PIK3CA mutations are commonly associated with most breast cancer subtypes. More recently PIK3CA mutations have been shown to induce tumor heterogeneity and are associated with activation of EGFR-signaling and reduced relapse free survival in basal subtype of breast cancer. Thus, understanding what determines PIK3CA induced heterogeneity and oncogenesis, is an important area of investigation. In this study, we assessed the effect of mutant PIK3CA together with mutant Ras plus mutant p53 on oncogenic behavior of two distinct stem/progenitor breast cell lines, designated as K5+/K19- and K5+/K19+. Constructs were ectopically overexpressed in K5+/K19- and K5+/K19+ stem/progenitor cells, followed by various in-vitro and in-vivo analyses. Oncogene combination m-Ras/m-p53/m-PIK3CA efficiently transformed both K5+/K19- and K5+/K19+ cell lines in-vitro, as assessed by anchorage-independent soft agar colony formation assay. Significantly, while this oncogene combination induced a complete epithelial-to-mesenchymal transition (EMT) in K5+/K19- cell line, mostly epithelial phenotype with minor EMT component was seen in K5+/K19+ cell line. However, both K5+/K19- and K5+/K19+ transformed cells exhibited increased invasion and migration abilities. Analyses of CD44 and CD24 expression showed both cell lines had tumor-initiating CD44+/CD24low cell population, however transformed K5+/K19- cells had more proportion of these cells. Significantly, both cell types exhibited in-vivo tumorigenesis, and maintained their EMT and epithelial nature in-vivo in mice tumors. Notably, while both cell types exhibited increase in tumor-initiating cell population, differential EMT phenotype was observed in these cell lines. These results suggest that EMT is a cell type dependent phenomenon and does not dictate oncogenesis.


[High and intensive care in long-term inpatient psychiatric services].
The increase in ambulant healthcare has also led to (simultaneously gained) insights that possibilities for temporary hospitalisation/admittance should be available for patients who experience a serious mental crisis. Based on experiences gained from the projects 'Reducing Coercion', high and intensive care (hic) has been developed. hic aims to improve care for psychiatric patients who are in need of crisis admission in a psychiatric hospital.
 AIM: To investigate whether the hic model is applicable to patients staying at a long-term psychiatric unit in hospital who, due to a severe psychiatric crisis, need to be hospitalised short-term in an emergency department of long-term inpatient care.
 METHOD: A responsive process evaluation in which three emergency departments with inpatient healthcare (from three different psychiatric institutions) participated. Starting point was application of the hic evaluator, followed by semi-structured interviews and focus groups.
 RESULTS: The items as described in the hic manual could be implemented within this setting. Stimulating and impeding factors were found, as well as adjustments needed to remove barriers to successful implementation.
 CONCLUSION: Although the hic model can be applied to the crisis departments of long-term inpatient psychiatric services, adjustments are indicated.


Building professionals' intention to use smart and sustainable building technologies - An empirical study.
Smart and sustainable buildings save energy and material resources and provide a comfortable environment that enhances their occupants' well-being and productivity. It is therefore crucial to understand how building professionals, including designers, engineers, and contractors, view smart and sustainable buildings and what drives them towards smart and sustainable building technologies. This study identifies salient smart and sustainable building features from building professionals' perspective and explores what determines building professionals' intention to use such building technologies. Responses from 543 Hong Kong's building professionals identify that intelligent security, intelligent and responsive fresh air supply, and intelligent and responsive thermal control are among the most important features of smart and sustainable buildings. Results of structural equation modeling grounded on an extended technology acceptance model indicate that facilitating condition and job relevance are related to perceived ease of use while subjective norm pertaining to image and perceived ease of use are predictors of perceived usefulness. Facilitating condition, perceived ease of use and perceived usefulness jointly influence building professionals' intention to use smart and sustainable building technologies.


Age-related morphometric changes of the tidemark in the ovine stifle.
Though the ovine stifle is commonly used to study osteoarthritis, there is limited information about the age-related morphometric changes of the tidemark. The objective of this study was to document the number of tidemarks in the stifle of research sheep without clinical signs of osteoarthritis and of various ages (n = 80). Articular cartilage of the medial and lateral tibial condyles and of the medial and lateral femoral condyles was assessed by histology: (a) to count the number of tidemark; and (b) to assess the OARSI (Osteoarthritis Research Society International) score for structural changes of cartilage. The number of tidemarks varied between anatomical regions, respectively, from 4.2 in the medial femoral condyle to 5.0 in the lateral tibial condyle. The axial part showed a significant higher number of tidemarks than the abaxial part, for all regions except the medial tibial condyle. Whilst the tidemark count strongly correlated with age (Spearman's correlation coefficient = 0.70; 95% confidence interval (95% CI): 0.67-0.73; p < 0.0001), the OARSI score was weakly correlated with age in our cohort of sheep (Spearman's correlation coefficient = 0.25; 95% CI: 0.19-0.30; p < 0.0001). Interestingly, no tidemark was seen in the three animals aged 6 months. Our data indicate that the number of tidemarks increases with age and vary with anatomical region. The regional variation also revealed a higher number of tidemarks in the tibia than in the femur. This could be attributed to the local variation in cartilage response to strain and to the difference in chondrocyte biology and density.


Gene expression mapping of histone deacetylases and co-factors, and correlation with survival time and 1H-HRMAS metabolomic profile in human gliomas.
Primary brain tumors are presently classified based on imaging and histopathological techniques, which remains unsatisfaying. We profiled here by quantitative real time PCR (qRT-PCR) the transcripts of eighteen histone deacetylases (HDACs) and a subset of transcriptional co-factors in non-tumoral brain samples from 15 patients operated for epilepsia and in brain tumor samples from 50 patients diagnosed with grade II oligodendrogliomas (ODII, n = 9), grade III oligodendrogliomas (ODIII, n = 22) and glioblastomas (GL, n = 19). Co-factor transcripts were significantly different in tumors as compared to non-tumoral samples and distinguished different molecular subgroups of brain tumors, regardless of tumor grade. Among all patients studied, the expression of HDAC1 and HDAC3 was inversely correlated with survival, whereas the expression of HDAC4, HDAC5, HDAC6, HDAC11 and SIRT1 was significantly and positively correlated with survival time of patients with gliomas. (1)H-HRMAS technology revealed metabolomically distinct groups according to the expression of HDAC1, HDAC4 and SIRT1, suggesting that these genes may play an important role in regulating brain tumorigenesis and cancer progression. Our study hence identified different molecular fingerprints for subgroups of histopathologically similar brain tumors that may enable the prediction of outcome based on the expression level of co-factor genes and could allow customization of treatment.


Targeting CpG oligonucleotides to the lymph node by nanoparticles elicits efficient antitumoral immunity.
Viral nucleic acids are recognized by specific pattern-recognition receptors of the Toll-like and RIG-I-like receptor families. Synthetic DNA and RNA oligonucleotides can activate the immune system through these receptors and potentiate Ab and CD8 cytotoxic responses to Ags. Systemic application of immunostimulatory oligonucleotides however also results in a generalized, non-Ag-specific stimulation of the immune system. In this study, we have dissociated the induction of an Ag-specific response from the systemic immune activation generally associated with immunostimulatory oligonucleotides. Delivery of CpG oligodeoxynucleotides that bind TLR9 by cationized gelatin-based nanoparticles potentiates the in vivo generation of an Ag-specific cytotoxic T cell and Ab response. Furthermore, immunization with CpG-loaded nanoparticles induces a protective antitumoral response in a murine model of melanoma. The systemic release of proinflammatory cytokines and widespread immunostimulation associated with free CpG is however completely abolished. In addition, we show that gelatin nanoparticle formulation prevents the destruction of lymphoid follicles mediated by CpG. Nanoparticle-delivered CpG, in contrast to free CpG, are selectively targeted to APCs in the lymph nodes where they mediate local immune stimulation. We describe a novel strategy to target immunostimulatory oligonucleotides to the initiation site of the immune response while at the same time protecting from an indiscriminate and generalized activation of the immune system.


In vitro developmental toxicity test detects inhibition of stem cell differentiation by silica nanoparticles.
While research into the potential toxic properties of nanomaterials is now increasing, the area of developmental toxicity has remained relatively uninvestigated. The embryonic stem cell test is an in vitro screening assay used to investigate the embryotoxic potential of chemicals by determining their ability to inhibit differentiation of embryonic stem cells into spontaneously contracting cardiomyocytes. Four well characterized silica nanoparticles of various sizes were used to investigate whether nanomaterials are capable of inhibition of differentiation in the embryonic stem cell test. Nanoparticle size distributions and dispersion characteristics were determined before and during incubation in the stem cell culture medium by means of transmission electron microscopy (TEM) and dynamic light scattering. Mouse embryonic stem cells were exposed to silica nanoparticles at concentrations ranging from 1 to 100 microg/ml. The embryonic stem cell test detected a concentration dependent inhibition of differentiation of stem cells into contracting cardiomyocytes by two silica nanoparticles of primary size 10 (TEM 11) and 30 (TEM 34) nm while two other particles of primary size 80 (TEM 34) and 400 (TEM 248) nm had no effect up to the highest concentration tested. Inhibition of differentiation of stem cells occurred below cytotoxic concentrations, indicating a specific effect of the particles on the differentiation of the embryonic stem cells. The impaired differentiation of stem cells by such widely used particles warrants further investigation into the potential of these nanoparticles to migrate into the uterus, placenta and embryo and their possible effects on embryogenesis.


Effect of denaturation of alpha-lactalbumin on the formation of BAMLET (bovine alpha-lactalbumin made lethal to tumor cells).
A complex of alpha-lactalbumin with oleic acid, also known as HAMLET/BAMLET (human/bovine alpha-lactalbumin made lethal to tumor cells), causes apoptosis-like death in tumor cells but has little effect on healthy differentiated cells. The aim of this study was to examine whether irreversible denaturation of alpha-lactalbumin is detrimental to the formation and cytotoxicity of BAMLET. Commercial bovine holo alpha-lactalbumin (1-4% w/v) was heated at 80 degrees C for up to 100 min. With an increasing concentration of protein, the denaturation of alpha-lactalbumin proceeded faster, and aggregation became more extensive. Native and sodium dodecyl sulfate polyacrylamide gel electrophoresis showed that a high proportion of the aggregates was linked by disulfide bonds. BAMLET was prepared from native and heat-treated alpha-lactalbumin according to a previously described chromatographic method. Despite the high content of denatured and aggregated alpha-lactalbumin in the heat-treated samples, their conversion into BAMLET was not negatively affected, resulting in BAMLET complexes partly composed of covalently linked aggregates of alpha-lactalbumin. The cytotoxicity of all prepared BAMLET samples was comparable to that of the control sample prepared from native alpha-lactalbumin (LD(50) = 34.6 +/- 2.7 mumol L(-1)). It was concluded that alpha-lactalbumin is not required to be in its native conformation for the conversion into its biologically active BAMLET complex.


Contrast nephropathy, pathophysiology and prevention.
Contrast nephropathy is a common cause of iatrogenic acute renal failure. Its incidence rises with the growing use of intra-arterial contrast in diagnostic and interventional procedures. Aim of the present review is to summarize the knowledge about pathophysiology and prevention. Nephrotoxicity is related to osmolality, dose and route of the contrast and only occurs in synergy with other factors, such as previous renal impairment and cardiovascular disease. With an interplay of these factors, contrast nephropathy has an impact on morbidity and mortality. Pathophysiological mechanisms are intrarenal vasoconstriction, leading to medullary ischemia, direct cytotoxicity, oxidative tissue damage and apoptosis. Several measures are of proven benefit in patients at risk. Among them are discontinuation of potentially nephrotoxic drugs, hydration, preferably with isotonic sodium bicarbonate, use of low osmolal contrast, oral or intravenous N-acetylcysteine and intravenous theophylline. In patients with severe cardiac and renal dysfunction undergoing cardiac interventions, periprocedural hemofiltration may be considered.


Anti-quorum sensing potential of Adenanthera pavonina.
Quorum sensing (QS) in Pseudomonas aeruginosa plays a key role in virulence factor production, biofilm formation and antimicrobial resistance. Because of emerging antimicrobial resistance in P. aeruginosa, there is a need to find an alternate nonantibiotic agent for the control of infections caused by this organism. To evaluate anti-QS activity of Adenanthera pavonina L., a medicinal plant used in traditional medicine. Preliminary screening for anti-QS activity of ethanol extract of A. pavonina was carried out using Chromobacterium violaceum CV026 biosensor strain and inhibition of QS-regulated violacein production was quantified using C. violaceum ATCC12472. Bioassay guided fractionation of ethanol extract resulted in ethyl acetate fraction (AEF) with strong anti-QS activity and AEF was evaluated for inhibition of QS-regulated pyocyanin production, proteolytic, elastolytic activity, swarming motility and biofilm formation in P. aeruginosa PAO1. AEF, at 0.5 mg/ml, inhibited pyocyanin production completely and at 1 mg/ml of AEF, complete inhibition of proteolytic and elastolytic activities were observed. However, viability of P. aeruginosa PAO1 was not affected at the tested concentrations of AEF as observed by cell count. Swarming motility was inhibited at the concentration of 0.1 mg/ml of AEF. Thin layer chromatography and biosensor overlay of AEF showed violacein inhibition zone at Rf value 0.63. From the results of this study, it can be concluded that A. pavonina extracts can be used as effective anti-QS agents.


Coadaptive aiding and automation enhance operator performance.
In this work, we expand on the theory of adaptive aiding by measuring the effectiveness of coadaptive aiding, wherein we explicitly allow for both system and user to adapt to each other. Adaptive aiding driven by psychophysiological monitoring has been demonstrated to be a highly effective means of controlling task allocation and system functioning. Psychophysiological monitoring is uniquely well suited for coadaptation, as malleable brain activity may be used as a continuous input to the adaptive system. To establish the efficacy of the coadaptive system, physiological activation of adaptation was directly compared with manual activation or no activation of the same automation and cuing systems. We used interface adaptations and automation that are plausible for real-world operations, presented in the context of a multi-remotely piloted aircraft control simulation. Each participant completed 3 days of testing during 1 week. Performance was assessed via proportion of targets successfully engaged. In the first 2 days of testing, there were no significant differences in performance between the conditions. However, in the third session, physiological adaptation produced the highest performance. By extending the data collection across multiple days, we offered enough time and repeated experience for user adaptation as well as online system adaptation, hence demonstrating coadaptive aiding. The results of this work may be employed to implement more effective adaptive workstations in a variety of work domains.


The mutant Asn291-->Ser human lipoprotein lipase is associated with reduced catalytic activity and does not influence binding to heparin.
Lipoprotein lipase (LPL) plays a central role in triglyceride metabolism, regulating the catabolism of triglyceride-rich lipoprotein particles. LPL performs its hydrolytic action attached to heparan sulfate proteoglycans at the luminal surface of capillary endothelial cells. We have assessed the effect of the Asn291-->Ser (N291S) substitution found in LPL gene from a human hyperlipemic patient. Our results showed that both the wild-type (WT) and N291S hLPL expressed in COS1 cells were secreted to the extracellular medium, and presented similar intracellular distribution patterns. Furthermore, heparin-Sepharose affinity chromatography assays revealed normal heparin affinity of the N291S hLPL. In addition, both the mutant and the WT protein bound to the surface of human fibroblasts and untransfected COS1 cells. Interestingly, diminished LPL specific activity was observed in the extracellular medium from mutant expressing cells. Therefore the lack of normal LPL activity in patients harbouring such a mutation could be the cause of their hyperlipemic disorder.


Different behavior of the resistance vessels of the human calf and forearm during contralateral isometric exercise, mental stress, and abnormal respiratory movements.
Experiments were conducted in normal human volunteers to compare the response of the forearm and calf vessels to contralateral isometric exercise, mental stress, resisted breathing, coughing, and the Valsalva maneuver. Blood flows were measured by means of strain-gauge plethysmography, arterial blood pressure by auscultation, and heart rate by electrocardiography. Isometric exercise of one forearm (at one-third maximal voluntary contraction) for 90 seconds caused an increase in blood pressure and heart rate; the vascular resistance decreased in the resting forearm, and increased in the calf. The decrease in forearm resistance was greater with the subjects supine and attenuated with the subjects standing or reclining head-down. With arterial occlusion of the exercising forearm just prior to cessation of the handgrip, the blood pressure and the calf resistance remained elevated, while the heart rate returned to control. The forearm resistance increased during the occlusion period and remained elevated throughout it. Mental stress caused an increase in heart rate and blood pressure and a dilation of the forearm but not of the calf vessels; these changes were smaller in standing than in supine subjects. Resisted breathing and coughing caused an increase in heart rate and in forearm blood flow, but not in calf blood flow. The Valsalva maneuver was followed by decreases in blood flow to the upper and lower limbs. The different responses in forearm and calf vessels can be explained by a central component which triggers a vasodilator pathway (possibly cholinergic) which is distributed to forearm but not to calf vessels.


Individual differences in allocation of funds in the dictator game associated with length of the arginine vasopressin 1a receptor RS3 promoter region and correlation between RS3 length and hippocampal mRNA.
Human altruism is a widespread phenomenon that puzzled evolutionary biologists since Darwin. Economic games illustrate human altruism by showing that behavior deviates from economic predictions of profit maximization. A game that most plainly shows this altruistic tendency is the Dictator Game. We hypothesized that human altruistic behavior is to some extent hardwired and that a likely candidate that may contribute to individual differences in altruistic behavior is the arginine vasopressin 1a (AVPR1a) receptor that in some mammals such as the vole has a profound impact on affiliative behaviors. In the current investigation, 203 male and female university students played an online version of the Dictator Game, for real money payoffs. All subjects and their parents were genotyped for AVPR1a RS1 and RS3 promoter-region repeat polymorphisms. Parents did not participate in online game playing. As variation in the length of a repetitive element in the vole AVPR1a promoter region is associated with differences in social behavior, we examined the relationship between RS1 and RS3 repeat length (base pairs) and allocation sums. Participants with short versions (308-325 bp) of the AVPR1a RS3 repeat allocated significantly (likelihood ratio = 14.75, P = 0.001, df = 2) fewer shekels to the 'other' than participants with long versions (327-343 bp). We also implemented a family-based association test, UNPHASED, to confirm and validate the correlation between the AVPR1a RS3 repeat and monetary allocations in the dictator game. Dictator game allocations were significantly associated with the RS3 repeat (global P value: likelihood ratio chi(2) = 11.73, df = 4, P = 0.019). The association between the AVPR1a RS3 repeat and altruism was also confirmed using two self-report scales (the Bardi-Schwartz Universalism and Benevolence Value-expressive Behavior scales). RS3 long alleles were associated with higher scores on both measures. Finally, long AVPR1a RS3 repeats were associated with higher AVPR1a human post-mortem hippocampal messenger RNA levels than short RS3 repeats (one-way analysis of variance (ANOVA): F = 15.04, P = 0.001, df = 14) suggesting a functional molecular genetic basis for the observation that participants with the long RS3 repeats allocate more money than participants with the short repeats. This is the first investigation showing that a common human polymorphism, with antecedents in lower mammals, contributes to decision making in an economic game. The finding that the same gene contributing to social bonding in lower animals also appears to operate similarly in human behavior suggests a common evolutionary mechanism.


Osteoblast-specific deletion of Pkd2 leads to low-turnover osteopenia and reduced bone marrow adiposity.
Polycystin-1 (Pkd1) interacts with polycystin-2 (Pkd2) to form an interdependent signaling complex. Selective deletion of Pkd1 in the osteoblast lineage reciprocally regulates osteoblastogenesis and adipogenesis. The role of Pkd2 in skeletal development has not been defined. To this end, we conditionally inactivated Pkd2 in mature osteoblasts by crossing Osteocalcin (Oc)-Cre;Pkd2+/null mice with floxed Pkd2 (Pkd2flox/flox) mice. Oc-Cre;Pkd2flox/null (Pkd2Oc-cKO) mice exhibited decreased bone mineral density, trabecular bone volume, cortical thickness, mineral apposition rate and impaired biomechanical properties of bone. Pkd2 deficiency resulted in diminished Runt-related transcription factor 2 (Runx2) expressions in bone and impaired osteoblastic differentiation ex vivo. Expression of osteoblast-related genes, including, Osteocalcin, Osteopontin, Bone sialoprotein (Bsp), Phosphate-regulating gene with homologies to endopeptidases on the X chromosome (Phex), Dentin matrix protein 1 (Dmp1), Sclerostin (Sost), and Fibroblast growth factor 23 (FGF23) were reduced proportionate to the reduction of Pkd2 gene dose in bone of Oc-Cre;Pkd2flox/+ and Oc-Cre;Pkd2flox/null mice. Loss of Pkd2 also resulted in diminished peroxisome proliferator-activated receptor γ (PPARγ) expression and reduced bone marrow fat in vivo and reduced adipogenesis in osteoblast culture ex vivo. Transcriptional co-activator with PDZ-binding motif (TAZ) and Yes-associated protein (YAP), reciprocally acting as co-activators and co-repressors of Runx2 and PPARγ, were decreased in bone of Oc-Cre;Pkd2flox/null mice. Thus, Pkd1 and Pkd2 have coordinate effects on osteoblast differentiation and opposite effects on adipogenesis, suggesting that Pkd1 and Pkd2 signaling pathways can have independent effects on mesenchymal lineage commitment in bone.


Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome.
A cell-line derived from a patient with chronic myelogenous leukemia (CML) is described. The new cell-line, which has over 175 serial passanges in a 3 1/2-yr period, has the following characteristics: (1) CML cells started to proliferate actively since they were first incubated in culture media. A threefold increase in the total number of cells was observed during the first seven passages; the cell population increased by a factor of 10 to 20 every 7 days from passage 8 through 85; from 20 to 40 times from passage 86 through 150, and more than 40 times after 150 passages. (2) The majority of the nononucleated cells are undifferentiated blasts. (3) The karyotype of all the cells examined show the Philadelphia (Ph1) chromosome and a long acrocentric marker plus aneuploidy. The Giemsa-banding studies identified the Ph1 chromosome as a terminal deletion of the long arm of chromosome 22:del(22)(q12) and the long acrocentric marker as an unbalanced reciprocal translocation of one chromosome 17 and the long arm of one chromosome 15. (4) The CML cells do not produce immunoglobulins, are free of mycoplasma, Epstein-Barr virus, and herpes-like virus particles. (5) CML cells have no alkaline phosphatase and myeloperoxidase activities and did not engulf inert particles. (6) Cultured CML cells provide a constant source of a specific antigen. This CML cell-line represents a unique source of CML cells with meaningful indicators of malignancy for clinical and experimental studies.


Biodistribution and Tumor Uptake of 67Ga-Nimotuzumab in a Malignant Pleural Mesothelioma Xenograft.
Malignant pleural mesothelioma (MPM) is the most common tumor of the pulmonary pleura. It is a rare and aggressive malignancy, generally associated with continuous occupational exposure to asbestos. Only a multimodal-approach to treatment, based on surgical resection, chemotherapy and/or radiation, has shown some benefits. However, the survival rate remains low. Nimotuzumab (h-R3), an anti-EGFR (epidermal growth factor receptor) humanized antibody, is proposed as a promising agent for the treatment of MPM. The aim of this research was to implement a procedure for nimotuzumab radiolabeling to evaluate its biodistribution and affinity for EGF (epidermal growth factor) receptors present in a mesothelioma xenograft. Nimotuzumab was radiolabeled with 67Ga; radiolabel efficiency, radiochemical purity, serum stability, and biodistribution were evaluated. Biodistribution and tumor uptake imaging studies by microSPECT/CT in mesothelioma xenografts revealed constant nimotuzumab uptake at the tumor site during the first 48 h after drug administration. In vivo studies using MPM xenografts showed a significant uptake of this radioimmunoconjugate, which illustrates its potential as a biomarker that could promote its theranostic use in patients with MPM.


General Anesthesia Causes Long-term Impairment of Mitochondrial Morphogenesis and Synaptic Transmission in Developing Rat Brain.
Clinically used general anesthetics, alone or in combination, are damaging to the developing mammalian brain. In addition to causing widespread apoptotic neurodegeneration in vulnerable brain regions, exposure to general anesthesia at the peak of synaptogenesis causes learning and memory deficiencies later in life. In vivo rodent studies have suggested that activation of the intrinsic (mitochondria-dependent) apoptotic pathway is the earliest warning sign of neuronal damage, suggesting that a disturbance in mitochondrial integrity and function could be the earliest triggering events. Because proper and timely mitochondrial morphogenesis is critical for brain development, the authors examined the long-term effects of a commonly used anesthesia combination (isoflurane, nitrous oxide, and midazolam) on the regional distribution, ultrastructural properties, and electron transport chain function of mitochondria, as well as synaptic neurotransmission, in the subiculum of rat pups. This anesthesia, administered at the peak of synaptogenesis, causes protracted injury to mitochondria, including significant enlargement of mitochondria (more than 30%, P < 0.05), impairment of their structural integrity, an approximately 28% increase in their complex IV activity (P < 0.05), and a twofold decrease in their regional distribution in presynaptic neuronal profiles (P < 0.05), where their presence is important for the normal development and functioning of synapses. Consequently, the authors showed that impaired mitochondrial morphogenesis is accompanied by heightened autophagic activity, decrease in mitochondrial density (approximately 27%, P < 0.05), and long-lasting disturbances in inhibitory synaptic neurotransmission. The interrelation of these phenomena remains to be established. Developing mitochondria are exquisitely vulnerable to general anesthesia and may be important early target of anesthesia-induced developmental neurodegeneration.


DNA synthesized in the hepatitis B Dane particle DNA polymerase reaction.
Radioactive DNA was prepared in extensive (4 h) Dane particle DNA polymerase reactions. In different experiments the amount of new DNA, determined by the amount of nucleotide incorporation into an acid-insoluble form, was between 29 and 45% of the total circular DNA isolated from Dane particle preparations after the reaction. DNA reassociation kinetics were used to determine the complexity of the newly synthesized DNA. In different experiments COt1/2 values, corresponding to between 625 and 1,250 nucleotide pairs, were obtained for the radioactive Dane particle DNA. These results suggest that a unique region (or regions), corresponsing to approximately one-fourth to one-half of the circular Dane particle DNA template, was copied one time during the reaction. DNA and RNA extracted from hepatitis B virus-infected liver but not from uninfected liver accelerated the rate of reassociation of radioactive DNA from Dane particles. These Dane particle DNA base sequences were found in alkali-stable, rapidly sedimenting DNA from infected liver as well as in DNA sedimenting at a rate similar to the DNA extracted from Dane particles. These findings are consistent with Dane particle DNA being hepatitis B virus DNA that is integrated into high-molecular-weight cellular DNA and transcribed into RNA in infected liver.


Child Health Assessment at School Entry (CHASE) project: evaluation in 10 London primary schools.
To assess the feasibility of implementing the Child Health Assessment at School Entry (CHASE) questionnaire, developed to capture the multiple dimensions of the health of children in their first year at school, and to evaluate data quality, reliability and validity. Parents of 278 year-1 children, from 10 primary schools in two London boroughs, received a parent questionnaire and school nurses completed a separate questionnaire from health and education records for children whose parents consented. Additional data on free school meal eligibility and ethnicity were obtained from the two Local Education Authorities. The parent questionnaire included the Strengths and Difficulties Questionnaire (SDQ) and four dimensions of the Child Health Questionnaire Parent Form-28 (CHQ-PF28). Response rate was 61%. The association between school free school meals eligibility and response rate in each school approached significance (r = -0.62, P = 0.05). Data completeness of the parent questionnaire was high (mean 98%). Data completeness of the school nurse questionnaire was more variable (mean 82%). Cronbach's Alpha was greater than 0.6 for four of the five SDQ scales and greater than 0.7 for the two CHQ-PF28 multi-item scales. Relative to parents with qualifications, parents with no qualifications rated their children as having significantly more conduct problems, peer problems, and overall mental health problems (P < 0.01) as assessed by the SDQ, and significantly lower global health (P < 0.01) as assessed by the CHQ-PF28. Children with special educational needs and children with long-standing illness or disability were rated as having significantly lower global health (P < 0.05) than children without these. Sample tables of inter-school and inter-borough comparison of key findings demonstrate considerable differences in physical and mental health status. The questionnaire was acceptable to parents and school nurses, and feasible to implement within existing school resources. Initial tests of internal reliability and validity are satisfactory. These data have the potential to inform interventions and service provision at school and borough level, and public health trends over time.