Characteristics of polymorphism at a VNTR locus 3' to the apolipoprotein B gene in five human populations.
We have analyzed the allele frequency distribution at the hypervariable locus 3' to the apolipoprotein B gene (ApoB 3' VNTR) in five well-defined human populations (Kacharis of northeast India, New Guinea Highlanders of Papua New Guinea, Dogrib Indians of Canada, Pehuenche Indians of Chile, and a relatively homogeneous Caucasian population of northern German extraction) by using the PCR technique. A total of 12 segregating alleles were detected in the pooled sample of 319 individuals. A fairly consistent bimodal pattern of allele frequency distribution, apparent in most of these geographically and genetically diverse populations, suggests that the ApoB 3' VNTR polymorphism predates the geographic dispersal of ancestral human populations. In spite of the observed high degree of polymorphism at this locus (expected heterozygosity levels 55%-78%), the genotype distributions in all populations (irrespective of their tribal or cosmopolitan nature) conform to their respective Hardy-Weinberg predictions. Furthermore, analysis of the congruence between expected heterozygosity and the observed number of alleles reveals that, in general, the allele frequency distributions at this locus are in agreement with the predictions of the classical mutation-drift models. The data also show that alleles that are shared by all populations have the highest average frequency within populations. These findings demonstrate the potential utility of highly informative hypervariable loci such as the ApoB 3' VNTR locus in population genetic research, as well as in forensic medicine and determination of biological relatedness of individuals.


Insight into N-terminal localization and dynamics of engineered virus-like particles.
Virus-like particles composed of the cowpea chlorotic mottle virus (CCMV) capsid protein (CP) have been extensively studied as carrier systems in nanoscience. One well-established method to improve their stability under physiological conditions is to fuse a stimulus-responsive elastin-like polypeptide (ELP) to the N-terminus of the CPs. Even though the N-terminus should in principle be localized in the inner cavity of the protein cage, studies on the native CCMV revealed its accessibility on the particle surface. We verified that such phenomenon also applies to ELP-CCMVs, by exploiting the covalent functionalization of the CP N-terminal domain via a sortase A-mediated reaction. Western-blot analysis and Förster resonance energy transfer (FRET) experiments furthermore revealed this to be caused by both the external display of the N-termini and the interchange of CPs among preformed capsids. Our findings demonstrate the tunability of ELP-CCMV stability and dynamics and their potential effect on the exploitation of such protein cages as a drug delivery system.


Supernumerary phantom limbs in spinal cord injury.
Case report and review of supernumerary phantom limbs in patients suffering from spinal cord injury (SCI). SCI rehabilitation centre. After a ski accident, a 71-year-old man suffered an incomplete SCI (level C3; AIS C, central cord syndrome), with a C3/C4 dislocation fracture. From the first week after injury, he experienced a phantom duplication of both upper limbs that lasted for 7 months. The supernumerary limbs were only occasionally related to painful sensation, specifically when they were perceived as crossed on his trunk. Although the painful sensations were responsive to pain medication, the presence of the illusory limb sensations were persistent. During neurological recovery, the supernumerary limbs gradually disappeared. A rubber hand illusion paradigm was used twice during recovery to monitor the patient's ability to integrate visual, tactile and proprioceptive stimuli. Overall, the clinical relevance of supernumerary phantom limbs is not clear, specific treatment protocols have not yet been developed, and the underlying neural mechanisms are not fully understood. Supernumerary phantom limbs have been previously reported in patients with (sub)cortical lesions, but might be rather undocumented in patients suffering from traumatic SCI. For the appropriate diagnosis and treatment after SCI, supernumerary phantoms should be distinguished from other phantom sensations and pain syndromes after SCI.


Antihypertensive treatment and plasma lipoprotein levels. The associations in data from a population study.
In clinical trials, antihypertensive therapy with beta blockers and methyldopa has resulted in adverse changes in high-density lipoprotein cholesterol levels. To investigate whether such effects can be observed in a general population in which beta blockers and methyldopa are prescribed widely for the treatment of high blood pressure, plasma lipid and lipoprotein levels were compared in subjects receiving antihypertensive treatment, subjects with untreated high blood pressure, and subjects with normal blood pressure; these subjects were drawn from a random sample of 5,603 men and women screened in a national study of risk-factor prevalence in Australia. In both sexes, high-density lipoprotein cholesterol levels were lower in the group receiving treatment for hypertension than in the other groups (p less than 0.001). High-density lipoprotein cholesterol levels in normotensive subjects and subjects with untreated high blood pressure were identical. In men, triglyceride levels and the ratio of total cholesterol to high-density lipoprotein cholesterol were higher in the group receiving treatment than in the other groups (p less than 0.001 and p less than 0.05, respectively). In both sexes, the differences in plasma lipids and lipoproteins between treated and untreated hypertensive groups were independent of age, body mass index, alcohol consumption, and smoking. More than 40 percent of untreated hypertensive subjects and more than 50 percent of treated hypertensive subjects had elevated total cholesterol levels (greater than or equal to 252 mg/dl) or an elevated ratio of total cholesterol to high-density lipoprotein cholesterol (greater than or equal to 6.0) in comparison with less than 30 percent of normotensive subjects. These results indicate that even without treatment, a large proportion of persons with high blood pressure had a significant abnormality in plasma cholesterol. The observation of a lower high-density lipoprotein cholesterol level in subjects receiving treatment for high blood pressure suggests that antihypertensive therapy as prescribed in this study population had worsened the already poor plasma lipid and lipoprotein profile of the average hypertensive patient.


Mediator and cohesin connect gene expression and chromatin architecture.
Transcription factors control cell-specific gene expression programs through interactions with diverse coactivators and the transcription apparatus. Gene activation may involve DNA loop formation between enhancer-bound transcription factors and the transcription apparatus at the core promoter, but this process is not well understood. Here we report that mediator and cohesin physically and functionally connect the enhancers and core promoters of active genes in murine embryonic stem cells. Mediator, a transcriptional coactivator, forms a complex with cohesin, which can form rings that connect two DNA segments. The cohesin-loading factor Nipbl is associated with mediator-cohesin complexes, providing a means to load cohesin at promoters. DNA looping is observed between the enhancers and promoters occupied by mediator and cohesin. Mediator and cohesin co-occupy different promoters in different cells, thus generating cell-type-specific DNA loops linked to the gene expression program of each cell.


Harsh childhood environmental characteristics predict exploitation and retaliation in humans.
Across and within societies, people vary in their propensities towards exploitative and retaliatory defection in potentially cooperative interaction. We hypothesized that this variation reflects adaptive responses to variation in cues during childhood that life will be harsh, unstable and short--cues that probabilistically indicate that it is in one's fitness interests to exploit co-operators and to retaliate quickly against defectors. Here, we show that childhood exposure to family neglect, conflict and violence, and to neighbourhood crime, were positively associated for men (but not women) with exploitation of an interaction partner and retaliatory defection after that partner began to defect. The associations between childhood environment and both forms of defection for men appeared to be mediated by participants' endorsement of a 'code of honour'. These results suggest that individual differences in mutual benefit cooperation are not merely due to genetic noise, random developmental variation or the operation of domain-general cultural learning mechanisms, but rather, might reflect the adaptive calibration of social strategies to local social-ecological conditions.


Urologic diseases in america project: benign prostatic hyperplasia.
Benign prostatic hyperplasia (BPH), the most common benign neoplasm in American men, is a chronic condition that is associated with progressive lower urinary tract symptoms and affects almost 3 of 4 men during the seventh decade of life. Approximately 6.5 million of the 27 million white men who are 50 to 79 years old in the United States in 2000 were estimated to meet the criteria for discussing treatment. The analytical methods used to generate these results have been described previously. In 2000 approximately 4.5 million visits were made to physician offices to for a primary diagnosis of BPH and almost 8 million visits were made with a primary or secondary diagnosis of BPH. In the same year approximately 87,400 prostatectomies for BPH were performed in inpatients in nonfederal hospitals in the United States. While the number of outpatient visits for BPH increased consistently during the 1990s, there was a dramatic decrease in the use of transurethral prostatectomy, inpatient hospitalization and length of hospital stay for this condition. These trends reflect the changing face of medical management for BPH, ie increasing use of pharmacological agents and minimally invasive therapies. In 2000 the direct cost of BPH treatment was estimated to be $1.1 billion exclusive of outpatient pharmaceuticals. Given the impact that BPH has on quality of life and health care cost in millions of American men, additional research into risk factors, diagnostic and therapeutic resource use, and effectiveness and cost benefit of therapies are warranted.


A carboxy-terminal fragment of protein mu 1/mu 1C is present in infectious subvirion particles of mammalian reoviruses and is proposed to have a role in penetration.
Penetration of a cell membrane as an early event in infection of cells by mammalian reoviruses appears to require a particular type of viral particle, the infectious subvirion particle (ISVP), which is generated from an intact virion by proteolytic cleavage of the outer capsid proteins sigma 3 and mu 1/mu 1C. Characterizations of the structural components and properties of ISVPs are thus relevant to attempts to understand the mechanism of penetration by reoviruses. In this study, a novel, approximately 13-kDa carboxy-terminal fragment (given the name phi) was found to be generated from protein mu 1/mu 1C during in vitro treatments of virions with trypsin or chymotrypsin to yield ISVPs. With trypsin treatment, both the carboxy-terminal fragment phi and the amino-terminal fragment mu 1 delta/delta were shown to be generated and to remain attached to ISVPs in stoichiometric quantities. Sites of protease cleavage were identified in the deduced amino acid sequence of mu 1 by determining the amino-terminal sequences of phi proteins: trypsin cleaves between arginine 584 and isoleucine 585, and chymotrypsin cleaves between tyrosine 581 and glycine 582. Findings in this study indicate that sequences in the phi portion of mu 1/mu 1C may participate in the unique functions attributed to ISVPs. Notably, the delta-phi cleavage junction was predicted to be flanked by a pair of long amphipathic alpha-helices. These amphipathic alpha-helices, together with the myristoyl group at the extreme amino terminus of mu 1/mu 1N, are proposed to interact directly with the lipid bilayer of a cell membrane during penetration by mammalian reoviruses.


Cyanidin and cyanidin 3-O-beta-D -glucoside as DNA cleavage protectors and antioxidants.
Anthocyanins, colored flavonoids, are water-soluble pigments present in the plant kingdom; in fact they are secondary plant metabolites responsible for the blue, purple, and red color of many plant tissues. Present in beans, fruits, vegetables and red wines, considerable amounts of anthocyanins are ingested as constituents of the human diet (180-215 mg daily). There is now increasing interest in the in vivo protective function of natural antioxidants contained in dietary plants against oxidative damage caused by free radical species. Recently, the antioxidant activity of phenolic phytochemicals, has been investigated. Since the antioxidant mechanism of anthocyanin pigments is still controversial, in the present study we evaluated the effects of cyanidin and cyanidin 3-O-beta-D-glucoside on DNA cleavage, on their free radical scavenging capacity and on xanthine oxidase activity. Cyanidin and cyanidin 3-O-beta-D-glucoside showed a protective effect on DNA cleavage, a dose-dependent free radical scavenging activity and significant inhibition of XO activity. These effects suggest that anthocyanins exhibit interesting antioxidant properties, and could therefore represent a promising class of compounds useful in the treatment of pathologies where free radical production plays a key role.


Cooperative DNA and histone binding by Uhrf2 links the two major repressive epigenetic pathways.
Gene expression is regulated by DNA as well as histone modifications but the crosstalk and mechanistic link between these epigenetic signals are still poorly understood. Here we investigate the multi-domain protein Uhrf2 that is similar to Uhrf1, an essential cofactor of maintenance DNA methylation. Binding assays demonstrate a cooperative interplay of Uhrf2 domains that induces preference for hemimethylated DNA, the substrate of maintenance methylation, and enhances binding to H3K9me3 heterochromatin marks. FRAP analyses revealed that localization and binding dynamics of Uhrf2 in vivo require an intact tandem Tudor domain and depend on H3K9 trimethylation but not on DNA methylation. Besides the cooperative DNA and histone binding that is characteristic for Uhrf2, we also found an opposite expression pattern of uhrf1 and uhrf2 during differentiation. While uhrf1 is mainly expressed in pluripotent stem cells, uhrf2 is upregulated during differentiation and highly expressed in differentiated mouse tissues. Ectopic expression of Uhrf2 in uhrf1(-/-) embryonic stem cells did not restore DNA methylation at major satellites indicating functional differences. We propose that the cooperative interplay of Uhrf2 domains may contribute to a tighter epigenetic control of gene expression in differentiated cells.


Comparative c-type cytochrome expression analysis in Shewanella oneidensis strain MR-1 and Anaeromyxobacter dehalogenans strain 2CP-C grown with soluble and insoluble oxidized metal electron acceptors.
The genomes of Shewanella oneidensis strain MR-1 and Anaeromyxobacter dehalogenans strain 2CP-C encode 40 and 69 putative c-type cytochrome genes respectively. Deletion mutant and biochemical studies have assigned specific functions to a few c-type cytochromes involved in electron transfer to oxidized metals in S. oneidensis strain MR-1. Although promising, the genetic approach is limited to gene deletions that produce a distinct phenotype and to an organism for which a genetic system is available. To investigate and compare c-type cytochrome expression in S. oneidensis strain MR-1 and Anaeromyxobacter dehalogenans strain 2CP-C more comprehensively, proteomic measurements were used to characterize lysates of cells grown with soluble Fe(III) (as ferric citrate) and insoluble Mn(IV) (as MnO2) as electron acceptors. Strain MR-1 expressed 19 and 20, and strain 2CP-C expressed 27 and 25, c-type cytochromes when grown with Fe(III) and Mn(IV) respectively. The majority of c-type cytochromes (77% for strain MR-1 and 63% for strain 2CP-C) were expressed under both growth conditions; however, the analysis also revealed unique c-type cytochromes that were specifically expressed in cells grown with soluble Fe(III) or insoluble Mn(IV). Proteomic characterization proved to be a promising approach for determining the c-type cytochrome complement expressed under different growth conditions, and will help to elucidate the specific functions of more c-type cytochromes that are the basis for Shewanella and Anaeromyxobacter respiratory versatility.


Nimbolide a limonoid from Azadirachta indica inhibits proliferation and induces apoptosis of human choriocarcinoma (BeWo) cells.
We investigated the cytotoxic effects of nimbolide, a limonoid present in leaves and flowers of the neem tree (Azadirachta indica) on human choriocarcinoma (BeWo) cells. Treatment with nimbolide resulted in dose- and time-dependent inhibition of growth of BeWo cells with IC(50) values of 2.01 and 1.19 microM for 7 and 24 h respectively, accompanied by downregulation of proliferating cell nuclear antigen. Examination of nuclear morphology revealed fragmentation and condensation indicating apoptosis. Increase in the generation of reactive oxygen species (ROS) that was reversed by addition of reduced glutathione suggested ROS involvement in the cytotoxicity of nimbolide. A decrease in Bcl-2/Bax ratio with increased expression of Apaf-1 and caspase-3, and cleavage of poly(ADP-ribose) polymerase provide compelling evidence that nimbolide-induced apoptosis is mediated by the mitochondrial pathway. The results of the present study suggest that nimbolide has immense potential in cancer prevention and therapy based on its antiproliferative and apoptosis inducing effects.


Increased gene sampling strengthens support for higher-level groups within leaf-mining moths and relatives (Lepidoptera: Gracillariidae).
Researchers conducting molecular phylogenetic studies are frequently faced with the decision of what to do when weak branch support is obtained for key nodes of importance. As one solution, the researcher may choose to sequence additional orthologous genes of appropriate evolutionary rate for the taxa in the study. However, generating large, complete data matrices can become increasingly difficult as the number of characters increases. A few empirical studies have shown that augmenting genes even for a subset of taxa can improve branch support. However, because each study differs in the number of characters and taxa, there is still a need for additional studies that examine whether incomplete sampling designs are likely to aid at increasing deep node resolution. We target Gracillariidae, a Cretaceous-age (~100 Ma) group of leaf-mining moths to test whether the strategy of adding genes for a subset of taxa can improve branch support for deep nodes. We initially sequenced ten genes (8,418 bp) for 57 taxa that represent the major lineages of Gracillariidae plus outgroups. After finding that many deep divergences remained weakly supported, we sequenced eleven additional genes (6,375 bp) for a 27-taxon subset. We then compared results from different data sets to assess whether one sampling design can be favored over another. The concatenated data set comprising all genes and all taxa and three other data sets of different taxon and gene sub-sampling design were analyzed with maximum likelihood. Each data set was subject to five different models and partitioning schemes of non-synonymous and synonymous changes. Statistical significance of non-monophyly was examined with the Approximately Unbiased (AU) test. Partial augmentation of genes led to high support for deep divergences, especially when non-synonymous changes were analyzed alone. Increasing the number of taxa without an increase in number of characters led to lower bootstrap support; increasing the number of characters without increasing the number of taxa generally increased bootstrap support. More than three-quarters of nodes were supported with bootstrap values greater than 80% when all taxa and genes were combined. Gracillariidae, Lithocolletinae + Leucanthiza, and Acrocercops and Parectopa groups were strongly supported in nearly every analysis. Gracillaria group was well supported in some analyses, but less so in others. We find strong evidence for the exclusion of Douglasiidae from Gracillarioidea sensu Davis and Robinson (1998). Our results strongly support the monophyly of a G.B.R.Y. clade, a group comprised of Gracillariidae + Bucculatricidae + Roeslerstammiidae + Yponomeutidae, when analyzed with non-synonymous changes only, but this group was frequently split when synonymous and non-synonymous substitutions were analyzed together. 1) Partially or fully augmenting a data set with more characters increased bootstrap support for particular deep nodes, and this increase was dramatic when non-synonymous changes were analyzed alone. Thus, the addition of sites that have low levels of saturation and compositional heterogeneity can greatly improve results. 2) Gracillarioidea, as defined by Davis and Robinson (1998), clearly do not include Douglasiidae, and changes to current classification will be required. 3) Gracillariidae were monophyletic in all analyses conducted, and nearly all species can be placed into one of six strongly supported clades though relationships among these remain unclear. 4) The difficulty in determining the phylogenetic placement of Bucculatricidae is probably attributable to compositional heterogeneity at the third codon position. From our tests for compositional heterogeneity and strong bootstrap values obtained when synonymous changes are excluded, we tentatively conclude that Bucculatricidae is closely related to Gracillariidae + Roeslerstammiidae + Yponomeutidae.


Paraaortic lymph node metastasis in patients with intra-abdominal malignancies: CT vs PET.
To compare the diagnostic accuracy of computed tomography (CT) and positron emission tomography (PET) for the preoperative detection of paraaortic lymph node (PAN) metastasis in patients with intra-abdominal malignancies. Sixty-six patients with intra-abdominal malignancies who underwent both CT and PET before lymphadenectomy were included in this study. Histopathologically, 13 patients had metastatic PAN, while 53 had non-metastatic PAN. The CT criteria for metastasis were: short diameter of > 8 mm, lobular or irregular shape, and/or combined ancillary findings, including necrosis, conglomeration, vessel encasement, and infiltration. The PET criterion was positive fluorodeoxyglucose (FDG) uptake. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of both modalities were compared with the pathologic findings, and the false positive and false negative cases with both CT and PET were analyzed. The sensitivity, specificity, PPV, NPV, and accuracy of CT were 61.5%, 84.9%, 50%, 90% and 80.3%, respectively. For PET, the percentages were 46.2%, 100%, 100%, 88.3%, and 89.4%. Additionally, there were 8 false positive CT cases (8/53, 15.1%) and zero false positive PET cases. Of the 13 metastatic PANs, there were 5 false negative CT scans (38.5%) and 7 (53.9%) false negative PET scans. For detecting PAN metastasis, CT is more sensitive than PET, while PET is more specific.


Exertional Hyponatremia and Serum Sodium Change During Ultraendurance Cycling.
Exertional hyponatremia (EH) during prolonged exercise involves all avenues of fluid-electrolyte gain and loss. Although previous research implicates retention of excess fluid, EH may involve either loss, gain, or no change of body mass. Thus, the etiology, predisposing factors, and recommendations for prevention are vague-except for advice to avoid excessive drinking. This retrospective field study presents case reports of two unacquainted recreational cyclists (LC, 31y and AM, 39 years) who began exercise with normal serum electrolytes but finished a summer 164-km ride (ambient, 34±5°C) with a serum [Na+] of 130 mmol/L. To clarify the etiology of EH, their pre- and post-exercise measurements were compared to a control group (CON) of 31 normonatremic cyclists (mean ± SD; 37±6 years; 141±3 mmol Na+/L). Anthropomorphic characteristics, exercise time, and post-exercise ratings of thermal sensation, perceived exertion and muscle cramp were similar for LC, AM and CON. These two hyponatremic cyclists consumed a large and similar volume of fluid (191 and 189 ml/kg), experienced an 11 mmol/L decrease of serum [Na+], reported low thirst sensations; however, LC gained 3.1 kg (+4.3% of body mass) during 8.9 hr of exercise and AM maintained body mass (+0.1kg, +0.1%, 10.6h). In the entire cohort (n = 33), post-event serum [Na+] was strongly correlated with total fluid intake (R2 = 0.45, p < .0001), and correlated moderately with dietary sodium intake (R2=0.28, p = .004) and body mass change (R2 = 0.22, p = .02). Linear regression analyses predicted the threshold of EH onset (<135 mmol Na+/L) as 168 ml fluid/kg. The wide range of serum [Na+] changes (+6 to -11 mmol/L) led us to recommend an individualized rehydration plan to athletes because the interactions of factors were complex and idiosyncratic.


What is the appropriate age cut-off for cycloplegia in refraction?
To investigate the age range for which cycloplegia provides additional information compared with non-cycloplegic refraction in teenagers and young adults. Data for 1295 subjects (704 female; 591 male) from the Twins Eye Study in Tasmania (TEST) and the Brisbane Adolescent Twin Study (mean age: 19.65 ± 3.56, range: 13-26 years) were included. For all participants, cycloplegia was induced by instillation of either one drop of 1% cyclopentolate (13-14 years) or one drop of 1% tropicamide (15-26 years). Pre- and postcycloplegic refractive errors for both eyes were measured using a Humphrey-598 automated refractor and spherical equivalents of refractive error were calculated. Generalized Estimating Equations (GEE) were used to model the spherical equivalent refraction (SER) for each eye against age (by year) and axial length (in the given eye). The mean group difference between pre- and postcycloplegic SER (post minus pre) was 0.17 ± 0.52 D and 0.12 ± 0.51 D for the right and left eyes, respectively, indicating that postcycloplegic refraction was generally more hyperopic/less myopic. The mean difference between pre- and postcycloplegic SER decreased from 0.36 ± 0.41 D in the 13-year-olds to 0.06 ± 0.50 D in people aged 25 years. After adjusting for family-relatedness, the difference between pre- and postcycloplegia SER was significant in all age groups up until the age of 20 years. Non-cycloplegic autorefraction can result in group mean SER differences of greater myopia than cycloplegic autorefraction and occurs in teenagers (13-19 years of age), but not in adults 20-26 years. These data suggest that cycloplegia is not required in population estimates of refractive error for young adults once they reach approximately 20 years of age.


Inverse Mie problem.
We apply functional analysis to the scattered electromagnetic field of a particle with spherical symmetry to obtain a pair of integral transforms for converting the Mie-scattering amplitudes S perpendicular (theta) and S parallel (theta) into the Mie coefficients an and bn. In the case of a homogeneous sphere, a simple mathematical construction is derived that uniquely inverts the Mie coefficients to find the refractive index and the radius of the particle. A more general method for construction of the refractive-index profile of an arbitrary sphere is discussed that follows from the treatment of Newton and Sabatier.


Assessment of the performance of triphenylphosphine for the voltammetric determination of elemental sulphur in cosmetic products.
In this paper, an alternative voltammetric method for the determination of elemental sulphur in cosmetic products is presented. It is based on the decrease of triphenylphosphine oxidation current in the presence of elemental sulphur by using a glassy carbon electrode. A solution of 2% (m/v) acetic acid and 0.6 mol L-1 sodium acetate in methanol was used as a supporting electrolyte. The experimental conditions for indirect determination of elemental sulphur were established. Using square-wave voltammetry, the analytical curve was linear in the elemental sulphur concentration range of 9.94-271 μmol L-1, with a detection limit of 2.59 μmol L-1. The method was successfully applied to determine elemental sulphur in soap bars and anti-acne cream, without any preliminary sample treatment, therefore, it is shortened and simplified. The results obtained with the indirect voltammetric method were not statistically different in comparison with a titrimetric one, at a 95% confidence level. Additionally, excellent recovery percentages were obtained, proving no matrix interferences.


Socio-Ecological Influences on Adolescent (Aged 10-17) Alcohol Use and Unhealthy Eating Behaviours: A Systematic Review and Synthesis of Qualitative Studies.
Excess body weight and risky alcohol consumption are two of the greatest contributors to global disease. Alcohol use contributes directly and indirectly to weight gain. Health behaviours cluster in adolescence and track to adulthood. This review identified and synthesised qualitative research to provide insight into common underlying factors influencing alcohol use and unhealthy eating behaviours amongst young people aged 10-17. Sixty two studies met inclusion criteria. Twenty eight studies focused on alcohol; 34 focused on eating behaviours. Informed by principles of thematic analysis and meta-ethnography, analysis yielded five themes: (1) use of alcohol and unhealthy food to overcome personal problems; (2) unhealthy eating and alcohol use as fun experiences; (3) food, but not alcohol, choices are based on taste; (4) control and restraint; and (5) demonstrating identity through alcohol and food choices. Young people faced pressure, reinforced by industry, to eat and drink in very specific ways, with clear social consequences if their attitudes or behaviour were deemed unacceptable. No qualitative studies were identified with an explicit and concurrent focus on adolescent eating behaviours and alcohol consumption. Further exploratory work is needed to examine the links between food and alcohol in young people's emotional, social and cultural lives.


Shared developmental gait disruptions across two mouse models of neurodevelopmental disorders.
Motor deficits such as abnormal gait are an underappreciated yet characteristic phenotype of many neurodevelopmental disorders (NDDs), including Williams Syndrome (WS) and Neurofibromatosis Type 1 (NF1). Compared to cognitive phenotypes, gait phenotypes are readily and comparably assessed in both humans and model organisms and are controlled by well-defined CNS circuits. Discovery of a common gait phenotype between NDDs might suggest shared cellular and molecular deficits and highlight simple outcome variables to potentially quantify longitudinal treatment efficacy in NDDs. We characterized gait using the DigiGait assay in two different murine NDD models: the complete deletion (CD) mouse, which models hemizygous loss of the complete WS locus, and the Nf1+/R681X mouse, which models a NF1 patient-derived heterozygous germline NF1 mutation. Longitudinal data were collected across four developmental time points (postnatal days 21-30) and one early adulthood time point. Compared to wildtype littermate controls, both models displayed markedly similar spatial, temporal, and postural gait abnormalities during development. Developing CD mice also displayed significant decreases in variability metrics. Multiple gait abnormalities observed across development in the Nf1+/R681X mice persisted into early adulthood, including increased stride length and decreased stride frequency, while developmental abnormalities in the CD model largely resolved by adulthood. These findings suggest that the subcomponents of gait affected in NDDs show overlap between disorders as well as some disorder-specific features, which may change over the course of development. Our incorporation of spatial, temporal, and postural gait measures also provides a template for gait characterization in other NDD models and a platform to examining circuits or longitudinal therapeutics.


Neurologic complications of cancer and cancer therapy.
When neurologic complications occur as a result of systemic cancer and cancer treatment, they can be more disabling for patients than their primary cancer and significantly impair functioning in varied domains. However, recognizing neurologic signs and symptoms as complications of cancer and its treatment can pose a challenge for healthcare providers. Oncology nurses must develop a high index of suspicion for neurologic complications when examining or interviewing patients who present with neurologic symptoms or deficits and have a known systemic cancer. The purpose of this article is to help oncology nurses identify the common presentations of those complications and understand the ways in which they occur, with the hope that early identification will facilitate appropriate medical intervention and slow the progression of neurologic deficits and systemic decline.


The discovery and development of inhaled therapeutics for migraine.
Migraine is a disabling primary headache disorder that requires effective treatments. Inhalation is currently being explored for the delivery of drugs for migraine. Pulmonary-route delivery of drugs shows potential advantages for its use as a treatment, particularly compared the oral route. Areas covered: The authors highlight the current state of the literature and review multiple therapies for migraine-utilizing inhalation as the route of administration. The following therapeutics are discussed: inhaled ergotamine, inhaled dihydroergotamine mesylate (MAP0004), inhaled prochlorperazine, and inhaled loxapine. Coverage is also given to normobaric oxygen, hyperbaric oxygen, and nitrous oxide therapies. Expert opinion: Inhalation of MAP0004 showed promising results in terms of efficacy for acute migraine treatment in phase 3 studies, together with a more favorable tolerability profile compared to parenteral dosing and a better pharmacokinetic profile versus oral or intranasal delivery. In phase 2 trials, inhaled prochlorperazine shows good pharmacokinetics and efficacy, in contrast to inhaled loxapine that did not provide encouraging results in terms of efficacy. The authors see the potential for the use of dihydroergotamine mesylate in clinical practice pending regulatory approval.


Protective role of dark soy sauce against acrylamide-induced neurotoxicity in rats by antioxidative activity.
Dark soy sauce (DSS) exerts antioxidant activity in vitro or in vivo. To test the possibility of chemoprevention by such dietary supplements on acrylamide (ACR)-induced subacute toxicity, Sprague-Dawley male rats were administered DSS dissolved in water (0.5 ml/kgbw x d) for 2 weeks after, before or during ACR exposure (0.2 mg/L in distillate water). DSS significantly improved the body weight gain, relative brain weights, and the gait abnormalities of ACR-treated rats when DSS treated at the same time as ACR-exposure (p < 0.05). DSS significantly improved the axonal degeneration, the ratio of myelinated nerves < 3 microm in diameter, degree of central chromatolysis of the ganglion neurons in peripheral nerves, and numbers of SYP (+) aberrant dots per mm cortex in the cerebellar molecular layer of ACR-treated rats no matter before, after, or during ACR-exposure (p < 0.05). DSS significantly decreased the malondialdehyde level and increased the superoxide dismutase activity in brain of ACR-treated rats when DSS treated during ACR-exposure (p < 0.05). These findings suggest that DSS plays a protective role against ACR-developed neurotoxicity and, partly at least, through an anti-oxidative mechanism. And it is worthy to note that DSS treatment at the same time as ACR exposure plays a more effective protective role than before or after ACR exposure.


Efficacy and safety of finasteride therapy for benign prostatic hyperplasia: results of a 2-year randomized controlled trial (the PROSPECT study). PROscar Safety Plus Efficacy Canadian Two year Study.
To evaluate the efficacy and safety of 2 years' treatment of moderate benign prostatic hyperplasia (BPH) with finasteride. Double-blind, parallel-group, placebo-controlled, multicentre, prospective randomized study. Outpatient care in 28 centres across Canada. Men aged 45 to 80, in good health, with moderate BPH and no evidence of prostate cancer. A total of 613 men were entered into the study; 472 completed the 2 years of treatment. After 1 month of receiving a placebo (run-in period), patients were given either finasteride (5 mg/d) or a placebo for 2 years. changes from baseline in BPH symptom scores, maximum urinary flow rates and prostate volume. onset, course and resolution of all adverse events during the treatment period. In the efficacy analyses the mean BPH symptom scores decreased 2.1 points (from 15.8 to 13.7) in the finasteride group, as compared with a decrease of 0.7 points (from 16.6 to 15.9) in the placebo group (P < or = 0.01). The maximum urinary flow rate increased by a mean of 1.4 mL/s (from 11.1 to 12.5 mL/s) in the finasteride group, as compared with an increase of 0.3 mL/s (from 10.9 to 11.2 mL/s) in the placebo group (p < or = 0.01). The mean prostate volume decreased by 21% (from a mean volume of 44.1 cm3 at baseline) in the treatment group; it increased by 8.4% (from a mean volume of 45.8 cm3 at baseline) in the placebo group (p < or = 0.01). In the safety analysis, the proportion of patients who experienced any adverse event was similar in the two groups (81.0% in the treatment group and 81.2% in the placebo group). However, the incidence of adverse events related to sexual dysfunction were significantly higher in the finasteride group than in the placebo group (ejaculation disorder 7.7% v. 1.7% and impotence 15.8% v. 6.3%; p < or = 0.01 for both parameters). Finasteride is a well-tolerated and effective alternative to watchful waiting in the treatment of moderate BPH.


Osteoclast-independent bone resorption by fibroblast-like cells.
To date, mesenchymal cells have only been associated with bone resorption indirectly, and it has been hypothesized that the degradation of bone is associated exclusively with specific functions of osteoclasts. Here we show, in aseptic prosthesis loosening, that aggressive fibroblasts at the bone surface actively contribute to bone resorption and that this is independent of osteoclasts. In two separate models (a severe combined immunodeficient mouse coimplantation model and a dentin pit formation assay), these cells produce signs of bone resorption that are similar to those in early osteoclastic resorption. In an animal model of aseptic prosthesis loosening (i.e. intracranially self-stimulated rats), it is shown that these fibroblasts acquire their ability to degrade bone early on in their differentiation. Upon stimulation, such fibroblasts readily release acidic components that lower the pH of their pericellular milieu. Through the use of specific inhibitors, pericellular acidification is shown to involve the action of vacuolar type ATPases. Although fibroblasts, as mesenchymal derived cells, are thought to be incapable of resorbing bone, the present study provides the first evidence to challenge this widely held belief. It is demonstrated that fibroblast-like cells, under pathological conditions, may not only enhance but also actively contribute to bone resorption. These cells should therefore be considered novel therapeutic targets in the treatment of bone destructive disorders.


Validation of optimal reference genes for quantitative real time PCR in muscle and adipose tissue for obesity and diabetes research.
The global incidence of obesity has led to an increasing need for understanding the molecular mechanisms that drive this epidemic and its comorbidities. Quantitative real-time RT-PCR (RT-qPCR) is the most reliable and widely used method for gene expression analysis. The selection of suitable reference genes (RGs) is critical for obtaining accurate gene expression information. The current study aimed to identify optimal RGs to perform quantitative transcriptomic analysis based on RT-qPCR for obesity and diabetes research, employing in vitro and mouse models, and human tissue samples. Using the ReFinder program we evaluated the stability of a total of 15 RGs. The impact of choosing the most suitable RGs versus less suitable RGs on RT-qPCR results was assessed. Optimal RGs differed between tissue and cell type, species, and experimental conditions. By employing different sets of RGs to normalize the mRNA expression of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α), we show that sub-optimal RGs can markedly alter the PGC1α gene expression profile. Our study demonstrates the importance of validating RGs prior to normalizing transcriptional expression levels of target genes and identifies optimal RG pairs for reliable RT-qPCR normalization in cells and in human and murine muscle and adipose tissue for obesity/diabetes research.


Degenerative changes following spinal fixation in a small animal model.
The objective of this study was to evaluate changes of the lumbar vertebral column following fixation. Using an established small animal (rat) model of spinal fixation (hypomobility), 3 contiguous lumbar segments (L4, L5, L6) were fixed with a specially engineered vertebral fixation device. Spinal segments of control rats were compared with those of animals with 1, 4, or 8 weeks of fixation. Subgroups of these fixation animals subsequently had the fixation device removed for 1, 2, 4, 8, or 12 weeks to evaluate the effects of attempting to reestablish normal forces to the vertebral segments following hypomobility. This Institutional Animal Care and Use Committee (IACUC) approved study was conducted in a university animal facility. Eighty-seven animals (23 controls animals and 64 fixation animals) were used in this study. Main Outcome Measures Outcome measures were degenerative changes of the vertebral bodies (VBs) and intervertebral disks (IVDs), zygapophysial (Z) joint osteophyte formation, and Z joint articular surface degeneration (ASD). Changes found in vertebral segments that were fixed (hypomobile) were compared with changes in adjacent nonfixed vertebral segments, and changes among fixation animals were compared with nonfixed controls. Main Results Very few degenerative changes were identified on the VBs and IVDs. Z joint changes were significant, both for osteophyte formation (analysis of variance [ANOVA], P <.0001) and ASD (ANOVA, P <.0001). Fixed segments had more degenerative changes than nonfixed segments for all Z joint parameters (ANOVA, P <.0001). Osteophyte formation and ASD were directly dependent on duration of fixation. These findings indicate that fixation (hypomobility) results in time-dependent degenerative changes of the Z joints.


Kallikrein 4 is expressed in malignant mesothelioma--further evidence for the histogenetic link between mesothelial and epithelial cells.
The objective of this study was to analyze Kallikrein 4 protein (hK4) expression in effusions and solid tumors of patients diagnosed with malignant mesothelioma (MM) and compare hK4 expression in MM with that in breast and ovarian adenocarcinomas. Sections from 65 MM (21 effusions, 44 solid tumors) and 63 breast carcinomas (28 effusions, 35 solid tumors) were stained for hK4 using immunohistochemistry. Results were compared with our previously published data for 284 ovarian carcinomas (181 effusions, 103 solid tumors). Expression of hK4 was detected in 26/65 (40%) MM and 52/63 (83%) breast carcinomas. Ovarian carcinoma showed staining values that were comparable to those in breast carcinoma (expression of hK4 in 144/181; 80% effusions and 85/103; 83% solid tumors). As opposed to our previous findings in ovarian carcinoma, hK4 expression was higher in solid tumors when compared with to effusions in both MM (P = 0.013) and breast carcinoma (P = 0.002). Comparative analysis of the three tumor types showed significantly higher expression in ovarian and breast adenocarcinomas when compared with MM (P < 0.001). In conclusion, hK4 is frequently expressed in MM, with higher levels detected in solid tumors, although its expression is more limited than in gynecological adenocarcinomas. The presence of hK4 in MM, a non-hormonally regulated tumor, provides further support to the histogenetic link between mesothelial and epithelial cells.


Extraordinary water adsorption characteristics of graphene oxide.
The laminated structure of graphene oxide (GO) confers unique interactions with water molecules which may be utilised in a range of applications that require materials with tuneable hygroscopic properties. The precise role of the expandable interlayer spacing and functional groups in GO laminates has not completely been understood to date. Herein, we report the experimental and theoretical investigations on the adsorption and desorption behaviour of water in GO laminates as a function of relative pressure. We observed that GO imparts high water uptake capacity of up to 0.58 gram of water per gram of GO (g g-1), which is significantly higher than silica gel as a conventional desiccant material. More interestingly, the adsorption and desorption kinetics of GO is five times higher than silica gel. The observed extraordinary adsorption/desorption rate can be attributed to the high capillary pressure in GO laminates as well as micro meter sized tunnel-like wrinkles located at the surface.


The effect of paraquat on the aerobic metabolism of rabbit alveolar macrophages and lung fibroblasts.
In this study the effects of paraquat on the aerobic metabolism and viability of isolated rabbit alveolar macrophages and lung fibroblasts were investigated, and compared with the effects of other known metabolic inhibitors, i.e. sodium fluoride (NaF) and potassium cyanide (KCN). The manometrically and polarographically determined endogenous oxygen consumption of lavaged alveolar macrophages compared very well (180,9 +/- 35,8 and 169,3 +/- 26,8 nmol per 10(6) viable cells per hour respectively). Exogenous glucose (10 mM) and autologous serum (1:3 v/v) added to the medium had no significant effect on the basal respiration rate. The mean cell protein content, determined by the micro-Kjeldahl and Lowry techniques, amounted to 242,6 +/- 37,6 microgram/10(6) macrophages. Paraquat (2 mM), like NaF (20mM) and KCN (5 mM), decreased the viability of the macrophages far less than it did the oxygen utilization of the viable cells, and resulted in an 80% inhibition of oxygen uptake. In contrast, paraquat (1 mM) induced a marked stimulation (230%) of the cyanide-insensitive respiration of alveolar macrophages. The concentrations of paraquat (nmol/10(3) cells) which reduce macrophage metabolism to almost zero were virtually non-toxic to fibroblasts, as measured by their oxygen consumption.


Atomistic modeling of a KRT35/KRT85 keratin dimer: folding in aqueous solution and unfolding under tensile load.
We present an atomistic model of a full KRT35/KRT85 dimer, a fundamental building block of human hair. For both monomers initial structures were generated using empirical tools based on homology considerations, followed by the formulation of a naiïve dimer model from docking the monomers in vacuum. Relaxation in aqueous solution was then explored from molecular dynamics simulation. Driven by hydrophobic segregation and protein-protein hydrogen bonding relaxation dynamics result in a folded dimer arrangement which shows a striking encounter of cystein groups. Our simulations hence suggests that (i) cystein groups in the coil regions of keratin are well suited to establish disulfide bonds between the two monomers that constitute the dimer, and (ii) the particularly large number of cystein groups in the head and tail regions promotes the connection of dimers to establish meso- to macroscale fibers. Moreover, we show the molecular mechanisms of elastic and plastic deformation under tensile load. Upon elongation beyond the elastic regime, unfolding was identified as the exposure of hydrophobic moieties and the breaking of protein-protein hydrogen bonds. Therein, the step-wise character of the series of unfolding events leads to a broad regime of constant force in response to further elongation.


Intrinsic atopic dermatitis shows similar TH2 and higher TH17 immune activation compared with extrinsic atopic dermatitis.
Atopic dermatitis (AD) is classified as extrinsic and intrinsic, representing approximately 80% and 20% of patients with the disease, respectively. Although sharing a similar clinical phenotype, only extrinsic AD is characterized by high serum IgE levels. Because most patients with AD exhibit high IgE levels, an "allergic"/IgE-mediated disease pathogenesis was hypothesized. However, current models associate AD with T-cell activation, particularly TH2/TH22 polarization, and epidermal barrier defects. We sought to define whether both variants share a common pathogenesis. We stratified 51 patients with severe AD into extrinsic AD (n = 42) and intrinsic AD (n = 9) groups (with similar mean disease activity/SCORAD scores) and analyzed the molecular and cellular skin pathology of lesional and nonlesional intrinsic AD and extrinsic AD by using gene expression (real-time PCR) and immunohistochemistry. A significant correlation between IgE levels and SCORAD scores (r = 0.76, P < 10(-5)) was found only in patients with extrinsic AD. Marked infiltrates of T cells and dendritic cells and corresponding epidermal alterations (keratin 16, Mki67, and S100A7/A8/A9) defined lesional skin of patients with both variants. However, higher activation of all inflammatory axes (including TH2) was detected in patients with intrinsic AD, particularly TH17 and TH22 cytokines. Positive correlations between TH17-related molecules and SCORAD scores were only found in patients with intrinsic AD, whereas only patients with extrinsic AD showed positive correlations between SCORAD scores and TH2 cytokine (IL-4 and IL-5) levels and negative correlations with differentiation products (loricrin and periplakin). Although differences in TH17 and TH22 activation exist between patients with intrinsic AD and those with extrinsic AD, we identified common disease-defining features of T-cell activation, production of polarized cytokines, and keratinocyte responses to immune products. Our data indicate that a TH2 bias is not the sole cause of high IgE levels in patients with extrinsic AD, with important implications for similar therapeutic interventions.


Use of glucometer and fasting blood glucose as screening tools for diabetes mellitus type 2 and glycated haemoglobin as clinical reference in rural community primary care settings of a middle income country.
Thailand is considered to be a middle income country, and to control and prevent type 2 diabetes mellitus (T2DM) is one of the main concerns of the Thai Ministry of Public Health (MoPH). Screening for T2DM and care for T2DM patients has been integrated into the primary health care system, especially in rural areas. The intention of this investigation is to link public health research at the academic level with the local health authorities of a district of a north-eastern province of the country. Epidemiological methods were applied to validate the screening tools fasting capillary blood glucose (CBG), measured by glucometer and venous blood for the determination of plasma glucose (VPG), used for screening for T2DM among asymptomatic villagers. For assessing the validity of these two methods glycated haemoglobin (HbA1c) values were determined and used as the 'clinical reference'. All together 669 villagers were investigated. Determinations of CBG and VPG resulted in suspected T2DM cases, with 7.3% when assessed by CBG and 6.4% by VPG using a cutoff point of 7 mmol/L (126 mg/dl). Taking HbA1c determinations with a cutoff point of 7% into account, the proportion of T2DM suspected participants increased to 10.4%. By estimating sensitivity, specificity and the positive predictive value of CBG and VPG against the 'clinical reference' of HbA1c, sensitivity below 50% for both screening methods has been observed. The positive predictive value was determined to be 58.5% for CBG and 56.8% for VPG. The specificity of the two screening tests was over 96%. The low sensitivity indicates that using fasting CBG or VPG as a screening tool in the field results in a high proportion of diseased individuals remaining undetected. The equally low positive predictive values (below 60%) indicate a high working load for the curative sector in investigating suspected T2DM cases to determine whether they are truly diseased or false positive cases according to the screening method. Further implications of the results and the controversial discussion related to the use of HbA1c as clinical evidence for suffering from T2DM are also discussed.


Success at first insemination in Australian Angus cattle: analysis of uncertain binary responses.
Field data from Australian Angus herds were used to investigate 2 methods of analyzing uncertain binary responses for success or failure at first insemination. A linear mixed model that included herd, year, and month of mating as fixed effects; unrelated service sire, additive animal, and residual as random effects; and linear and quadratic effects of age at mating as covariates was used to analyze binary data. An average gestation length (GL) derived from artificial insemination data was used to assign an insemination date to females mated to natural service sires. Females that deviated from this average GL led to uncertain binary responses. Two analyses were carried out: 1) a threshold model fitted to uncertain binary data, ignoring uncertainty (M1); and 2) a threshold model fitted to uncertain binary data, accounting for uncertainty via fuzzy logic classification (M2). There was practically no difference between point estimates obtained from M1 and M2 for service sire and herd variance; however, when uncertain binary data were analyzed ignoring uncertainty (M1), additive variance and heritability estimates were greater than with M2. Pearson correlations indicated that no major reranking would be expected for service sire effects and animal breeding values using M1 and M2. Given the results of the current study, a threshold model contemplating uncertainty is suggested for noisy binary data to avoid bias when estimating genetic parameters.


Differences in information seeking among breast, prostate, and colorectal cancer patients: results from a population-based survey.
There is much research describing cancer patients' information needs and their use of the Internet, print media, and other sources to fulfill these needs. Yet little is known about whether patients with different types of cancer vary in their information needs and seeking behaviors. This study used population-based data to address this question. A sample was randomly drawn from the list of patients with breast, prostate, or colorectal cancer reported to the Pennsylvania Cancer Registry in 2005. Patients completed a mail survey (N=2010); respective response rates were 68%, 64%, and 61%. Colorectal cancer patients reported consistently less information seeking than breast and prostate cancer patients. Multivariate analyses revealed that differences by cancer type were not explained by sex or other demographics, disease stage, or treatment received. These differences were most pronounced among patients with early stage cancer. Cancer patients have myriad information needs and use a range of sources to satisfy these needs, but there appear to be important differences in information engagement by cancer type. Understanding differences in information seeking among disease-specific populations may help guide future patient education and decision making across the care continuum.


A herpes simplex virus 1 (McKrae) mutant lacking the glycoprotein K gene is unable to infect via neuronal axons and egress from neuronal cell bodies.
We have shown that the herpes simplex virus 1 (HSV-1) gK gene is essential for efficient replication and spread in the corneal epithelium and trigeminal ganglion neuroinvasion in mice (A. T. David, A. Baghian, T. P. Foster, V. N. Chouljenko, and K. G. Kousoulas, Curr. Eye Res. 33:455-467, 2008). To further investigate the role of gK in neuronal infection, we utilized a microfluidic chamber system separating neuronal cell bodies and axonal termini. HSV-1 (McKrae) engineered virus constitutively expressing enhanced green fluorescence protein (GFP) was efficiently transmitted in both a retrograde and an anterograde manner. These results were corroborated by expression of virion structural proteins in either chamber, as well as detection of viral genomes and infectious viruses. In contrast, efficient infection of either chamber with a gK-null virus did not result in infection of the apposed chamber. These results show that gK is an important determinant in virion axonal infection. Moreover, the inability of the gK-null virus to be transmitted in an anterograde manner suggests that virions acquire cytoplasmic envelopes prior to entering axons. Herpes simplex virus 1 (HSV-1) enters mucosal epithelial cells and neurons via fusion of the viral envelope with cellular membranes, mediated by viral glycoprotein B (gB) in cooperation with other viral glycoproteins. Retrograde transport of virions to neuronal cell bodies (somata) establishes lifelong latent infection in ganglionic neurons. We have previously reported that gK binds gB and is required for gB-mediated membrane fusion (Jambunatathan et al., J. Virol. 85:12910-12918, 2011; V. N. Chouljenko, A. V. Iyer, S. Chowdhury, J. Kim, and K. G. Kousoulas, J. Virol. 84:8596-8606, 2010). In the current study, we constructed a recombinant virus with the gK gene deleted in the highly virulent ocular HSV-1 strain McKrae. This recombinant virus failed to infect rat ganglionic neuronal axons alone or cocultured with Vero cells in microfluidic chambers. In addition, lack of gK expression prevented anterograde transmission of virions. These results suggest that gK is a critical determinant for neuronal infection and transmission.


Stepwise copper(i) binding to metallothionein: a mixed cooperative and non-cooperative mechanism for all 20 copper ions.
Copper is a ubiquitous trace metal of vital importance in that it serves as a cofactor in many metalloenzymes. Excess copper becomes harmful if not sequestered appropriately in the cell. As a metal ion chaperone, metallothionein (MT) has been proposed as a key player in zinc and copper homeostasis within the cell. The underlying mechanisms by which MT sequesters and transfers copper ions, and subsequently achieves its proposed biological function remain unknown. Using a combination of electrospray ionization mass spectrometry (ESI-MS), circular dichroism (CD), and emission spectroscopy, we report that the Cu(i) to human apo-MT1a binding mechanism is highly pH-dependent. The 20 relative Kf-values for the binding of 1 to 20 Cu(i) to the 20 cysteines of MT were obtained from computational simulation of the experimental mass spectral results. These data identified the pH-dependent formation of three sequential but completely different Cu-SCYS clusters, as a function of Cu(i) loading. These data provide the first overall sequence for Cu(i) binding in terms of domain specificity and transient binding site structures. Under cooperative binding at pH 7.4, a series of four clusters form: Cu4SCYS-6, followed by Cu6SCYS-9 (β), then a second Cu4SCYS-6 (α), and finally Cu7SCYS-x (α) (x = up to 11). Upon further addition of Cu(i), a mixture of species is formed in a non-cooperative mechanism, saturating the 20 cysteines of MT1a. Using benzoquinone, a cysteine modifier, we were able to confirm that Cu6SCYS-9 formed solely in the N-terminal β-domain, as well as confirming the existence of the presumed Cu4SCYS-6 cluster in the α-domain. Based on the results of ESI-MS and computational simulation we were able to identify Cu:MT speciation that resulted in specific emission and CD spectral properties.


Genome-wide analysis of Italian sheep diversity reveals a strong geographic pattern and cryptic relationships between breeds.
Italy counts several sheep breeds, arisen over centuries as a consequence of ancient and recent genetic and demographic events. To finely reconstruct genetic structure and relationships between Italian sheep, 496 subjects from 19 breeds were typed at 50K single nucleotide polymorphism loci. A subset of foreign breeds from the Sheep HapMap dataset was also included in the analyses. Genetic distances (as visualized either in a network or in a multidimensional scaling analysis of identical by state distances) closely reflected geographic proximity between breeds, with a clear north-south gradient, likely because of high levels of past gene flow and admixture all along the peninsula. Sardinian breeds diverged more from other breeds, a probable consequence of the combined effect of ancient sporadic introgression of feral mouflon and long-lasting genetic isolation from continental sheep populations. The study allowed the detection of previously undocumented episodes of recent introgression (Delle Langhe into the endangered Altamurana breed) as well as signatures of known, or claimed, historical introgression (Merino into Sopravissana and Gentile di Puglia; Bergamasca into Fabrianese, Appenninica and, to a lesser extent, Leccese). Arguments that would question, from a genomic point of view, the current breed classification of Bergamasca and Biellese into two separate breeds are presented. Finally, a role for traditional transhumance practices in shaping the genetic makeup of Alpine sheep breeds is proposed. The study represents the first exhaustive analysis of Italian sheep diversity in an European context, and it bridges the gap in the previous HapMap panel between Western Mediterranean and Swiss breeds.


["Atypical" method for understanding dementia. How can studying epigenetics contribute?].
The pathological hallmark of neurodegeneration is presence of intra- and extra neuronal inclusion bodies such as Lewy bodies in Parkinson's disease, senile plaques and neurofibrillary tangles in Alzheimer's disease. These are consisted of aggregated conformationally abnormal proteins. The precise mechanism of aggregation remains unknown, but increased expression of aggregation-prone proteins can lead to their aggregation. For example, in Down syndrome, duplication of the 21(st) chromosome, which contains the amyloid beta precursor protein (APP) gene, leads to accumulation of amyloid beta and Alzheimer's disease pathology and multiplication of APP gene is shown to be the cause of familial Alzheimer's disease. Moreover, in rare cases of PD, duplication or triplication of SNCA gene leads to alpha-synuclein accumulation, with triplication producing a more severe phenotype than duplication, suggesting that SNCA expression level determines the severity of the pathology. Lastly, animal models of neurodegenerative disorders are generated by over-expression of causal genes, further supporting the conclusion that increased gene expression is related to pathogenesis. Additional evidence indicates that SNCA promoter polymorphisms increases alpha-synuclein expression and increases susceptibility to sporadic PD. In addition to promoter polymorphisms, epigenetic modification can alter downstream gene expression. Epigenetic regulation includes histone modification and DNA methylation, of which CpG island methylation can be gene-specific; in several different cancers, CpG methylation inhibits binding of the transcription machinery, causing silencing of a specific oncogene, which leads to carcinogenesis. In central nervous system disorders, CpG methylation has been associated with psychiatric disorders, such as autism and schizophrenia. We found several cases of Parkinson's disease with epigenetic abnormality in SNCA gene. Thus, we believe that studying epigenetics can provide previously unknown causes for dementia and other neurodegenerative disorders.


Olfactory representations by Drosophila mushroom body neurons.
Learning and memory has been studied extensively in Drosophila using behavioral, molecular, and genetic approaches. These studies have identified the mushroom body as essential for the formation and retrieval of olfactory memories. We investigated odor responses of the principal neurons of the mushroom body, the Kenyon cells (KCs), in Drosophila using whole cell recordings in vivo. KC responses to odors were highly selective and, thus sparse, compared with those of their direct inputs, the antennal lobe projection neurons (PNs). We examined the mechanisms that might underlie this transformation and identified at least three contributing factors: excitatory synaptic potentials (from PNs) decay rapidly, curtailing temporal integration, PN convergence onto individual KCs is low ( approximately 10 PNs per KC on average), and KC firing thresholds are high. Sparse activity is thought to be useful in structures involved in memory in part because sparseness tends to reduce representation overlaps. By comparing activity patterns evoked by the same odors across olfactory receptor neurons and across KCs, we show that representations of different odors do indeed become less correlated as they progress through the olfactory system.


Reporting nuclear cardiology: a joint position paper by the European Association of Nuclear Medicine (EANM) and the European Association of Cardiovascular Imaging (EACVI).
The report of an imaging procedure is a critical component of an examination, being the final and often the only communication from the interpreting physician to the referring or treating physician. Very limited evidence and few recommendations or guidelines on reporting imaging studies are available; therefore, an European position statement on how to report nuclear cardiology might be useful. The current paper combines the limited existing evidence with expert consensus, previously published recommendations as well as current clinical practices. For all the applications discussed in this paper (myocardial perfusion, viability, innervation, and function as acquired by single photon emission computed tomography and positron emission tomography or hybrid imaging), headings cover laboratory and patient demographics, clinical indication, tracer administration and image acquisition, findings, and conclusion of the report. The statement also discusses recommended terminology in nuclear cardiology, image display, and preliminary reports. It is hoped that this statement may lead to more attention to create well-written and standardized nuclear cardiology reports and eventually lead to improved clinical outcome.


Systematic functional regulatory assessment of disease-associated variants.
Genome-wide association studies have discovered many genetic loci associated with disease traits, but the functional molecular basis of these associations is often unresolved. Genome-wide regulatory and gene expression profiles measured across individuals and diseases reflect downstream effects of genetic variation and may allow for functional assessment of disease-associated loci. Here, we present a unique approach for systematic integration of genetic disease associations, transcription factor binding among individuals, and gene expression data to assess the functional consequences of variants associated with hundreds of human diseases. In an analysis of genome-wide binding profiles of NFκB, we find that disease-associated SNPs are enriched in NFκB binding regions overall, and specifically for inflammatory-mediated diseases, such as asthma, rheumatoid arthritis, and coronary artery disease. Using genome-wide variation in transcription factor-binding data, we find that NFκB binding is often correlated with disease-associated variants in a genotype-specific and allele-specific manner. Furthermore, we show that this binding variation is often related to expression of nearby genes, which are also found to have altered expression in independent profiling of the variant-associated disease condition. Thus, using this integrative approach, we provide a unique means to assign putative function to many disease-associated SNPs.


1,1'-(9-Octyl-9H-carbazole-3,6-di-yl)diethanone.
The central structural element of the title compound, C(24)H(29)NO(2), is a carbazole unit substituted with two acetyl residues and an octyl chain. The acetyl residues are nearly coplanar [dihedral angles = 5.37 (14) and 1.0 (3)°] with the carbazole unit which is essentially planar (r.m.s. deviation for all non-H atoms = 0.025 Å). The octyl chain adopts an all-trans conformation. The crystal packing is stabilized by C-H⋯O hydrogen bonds.


Stem cell transplantation for autoimmune diseases.
Much progress has been made in the field of haemopoietic stem cell transplants (HSCTs) for severe autoimmune disorders. Theoretical considerations, animal data and anecdotal evidence suggested some time ago that intensive immunoablation followed by autologous HSCT could restore normal immune reactivity in patients with severe autoimmune disorders. Based on a concept statement issued in 1995, two European societies, the European League Against Rheumatism (EULAR) and the European Group for Blood and Marrow Transplantation (EBMT) began collecting phase I/II trial data in an international collaborative network. Sufficient information from more than 350 patients allows a preliminary assessment with level three evidence. Autologous HSCTs can induce remissions in all disease categories tested so far. Remissions can be transient or durable. HSCTs are associated with significant morbidity and mortality. Treatment-related mortality (TRM) is near 10% at 1 year and is associated with the intensity of the conditioning and the stage of the disease at the time of transplant. Marked interdisease differences exist. There are few data available in haematological autoimmune diseases, more in systemic sclerosis (SSc), systemic lupus erythematosus (SLE), juvenile idiopathic arthritis (JIA) and multiple sclerosis (MS). Patient selection has been recognized as a crucial element from the phase I-II trials. Patients with advanced disease, severely compromised organ function or irreversible organ damage should not be considered as candidates for HSCT. Prospective randomized studies should now determine the value of HSCT compared to standard therapy. Such trials are ongoing for patients with systemic sclerosis (ASTIS trial--Autologous Stem Cell Transplantation International Scleroderma Trial) or are planned for patients with multiple sclerosis (ASTIMS trial--Autologous Stem Cell Transplantation International Multiple Sclerosis Trial) and rheumatoid arthritis (ASTIRA trial--Autologous Stem Cell Transplantation International Rheumatoid Arthritis Trial). More phase II data are needed for other indications such as SLE and JIA.


International medical graduates in South Africa and the implications of addressing the current surgical workforce shortage.
The surgical workforce in South Africa is currently insufficient in being able to meet the burden of surgical disease in the country. International medical graduates (IMGs) help to alleviate the deficit, yet very little is known about these doctors and their career progression in our healthcare system. The demographic profile and career progression of IMGs who worked in our surgical department in a major university hospital in South Africa was reviewed over a four-year period. Twenty-eight IMGs were identified. There were 23 males (92%) and their mean age was 33 years. Seventy-one per cent (20/28) were on a fixed-term service contract, and returned to their respective country of origin. The option of renewing their service contracts was available to the 16 IMGs who left. Three explicitly indicated they would have stayed in South Africa if formal training was possible. Eight of the 28 IMGs (29%) extended their tenure, and remained in the service position as medical officers. All of the eight IMGs stayed with the intention of entering a formal surgical training programme. IMGs represented a significant proportion of service provision in our unit. Over one third of IMGs stayed beyond their initial tenure, and of these, all stayed in order to gain entry into the formal surgical training programme. A significant proportion of those who left would have stayed if entry to the programme was feasible.


Quantification of the Light Subunit of Neurofilament Protein in Cerebrospinal Fluid of Huntington's Disease Patients.
Neurofilament light proteins (NFL) are a structural element of the neuronal cytoskeleton and are released with neuronal damage. Its levels are increased in cerebrospinal fluid (CSF) in the setting of neurodegenerative diseases. We investigated the CSF-NFL levels of Huntington´s disease (HD) patients (participating in a clinical trial SAT-HD) as well as of premanifest carriers and compared their results with a sample of healthy controls and correlated CSF-NFL levels with demographic and clinical variables (baseline demographic characteristics and HD measures of disease severity). CSF levels were significantly higher in all HD subjects [5014.4 (1557.3) ng/l] and pre-manifest carriers [1050 (212.13) ng/l as compared to controls [331.4 (200.2) ng/l] (p<0.00) and were correlated with age (correlation coefficient -0.37, p<0.01) and CAG triplet number (0,51, p<0.05) in the subset of HD patients. NFL levels were not correlated with age in the control group. We did not find any correlation with the remaining variables. These results indicate, as in previous studies, that CSF-NFL levels are a marker of neuronal damage in HD. It seems to be a highly sensitive, but non-specific marker of axonal damage. One of the limitations of our study is a very small number of patients in pre-symptomatic group and lack of individuals with very advanced HD. Further investigations should focus on study of CSF-NFL levels in advanced patients, tracking prospectively CSF-NFL levels and analysing its correlation with the clinical course and usefulness to monitor disease progression, validation and quantification of NFL levels in more accessible biofluids.


The molecular scaffold KSR1 regulates the proliferative and oncogenic potential of cells.
The specificity of signaling through mitogen-activated protein kinase pathways has been attributed to both the control of intensity and duration of signaling and the actions of protein scaffolds. Here we demonstrate that the molecular scaffold KSR1 regulates the intensity and duration of ERK activation to modulate a cell's proliferative and oncogenic potential. Deletion of KSR1 eliminates the prolonged phase of ERK activation induced by platelet-derived growth factor and blocks Ras(V12)-induced transformation. The introduction of KSR1 into KSR1(-/-) mouse embryo fibroblasts causes a concentration-dependent increase in signaling and transformation, to a maximum at 14 times the wild-type KSR1 expression levels, but inhibits these responses at higher expression levels. An increase in KSR1 expression to levels that are optimal for signaling leads to a threefold increase in proliferative capacity and is coincident with the level of KSR1 expression that maximally associates with all members of the Raf/MEK/ERK cascade. These data reveal that cells contain a reserve proliferative capacity that is accessible by the optimal expression of a noncatalytic signaling component and that altering the expression level of a molecular scaffold can modulate the actions of growth factors and oncogenes.


Metaxa: a software tool for automated detection and discrimination among ribosomal small subunit (12S/16S/18S) sequences of archaea, bacteria, eukaryotes, mitochondria, and chloroplasts in metagenomes and environmental sequencing datasets.
The ribosomal small subunit (SSU) rRNA gene has emerged as an important genetic marker for taxonomic identification in environmental sequencing datasets. In addition to being present in the nucleus of eukaryotes and the core genome of prokaryotes, the gene is also found in the mitochondria of eukaryotes and in the chloroplasts of photosynthetic eukaryotes. These three sets of genes are conceptually paralogous and should in most situations not be aligned and analyzed jointly. To identify the origin of SSU sequences in complex sequence datasets has hitherto been a time-consuming and largely manual undertaking. However, the present study introduces Metaxa ( http://microbiology.se/software/metaxa/ ), an automated software tool to extract full-length and partial SSU sequences from larger sequence datasets and assign them to an archaeal, bacterial, nuclear eukaryote, mitochondrial, or chloroplast origin. Using data from reference databases and from full-length organelle and organism genomes, we show that Metaxa detects and scores SSU sequences for origin with very low proportions of false positives and negatives. We believe that this tool will be useful in microbial and evolutionary ecology as well as in metagenomics.


Three-station three-dimensional bolus-chase MR angiography with real-time fluoroscopic tracking.
To determine the feasibility of using real-time fluoroscopic tracking for bolus-chase magnetic resonance (MR) angiography of peripheral vasculature to image three stations from the aortoiliac bifurcation to the pedal arteries. This prospective study was institutional review board approved and HIPAA compliant. Eight healthy volunteers (three men; mean age, 48 years; age range, 30-81 years) and 13 patients suspected of having peripheral arterial disease (five men; mean age, 67 years; age range, 47-81 years) were enrolled and provided informed consent. All subjects were imaged with the fluoroscopic tracking MR angiographic protocol. Ten patients also underwent a clinical computed tomographic (CT) angiographic runoff examination. Two readers scored the MR angiographic studies for vessel signal intensity and sharpness and presence of confounding artifacts and venous contamination at 35 arterial segments. Mean aggregate scores were assessed. The paired MR angiographic and CT angiographic studies also were scored for visualization of disease, reader confidence, and overall diagnostic quality and were compared by using a Wilcoxon signed rank test. Real-time fluoroscopic tracking performed well technically in all studies. Vessel segments were scored good to excellent in all but the following categories: For vessel signal intensity and sharpness, the abdominal aorta, iliac arteries, distal plantar arteries, and plantar arch were scored as fair to good; and for presence of confounding artifacts, the abdominal aorta and iliac arteries were scored as fair. The MR angiograms and CT angiograms did not differ significantly in any scoring category (reader 1: P = .50, .39, and .39; reader 2: P = .41, .61, and .33, respectively). CT scores were substantially better in 20% (four of 20) and 25% (five of 20) of the pooled evaluations for the visualization of disease and overall image quality categories, respectively, versus 5% (one of 20) for MR scores in both categories. Three-station bolus-chase MR angiography with real-time fluoroscopic tracking provided high-spatial-resolution arteriograms of the peripheral vasculature, enabled precise triggering of table motion, and compared well with CT angiograms.


Does suboptimal household flooring increase the risk of diarrhoea and intestinal parasite infection in low and middle income endemic settings? A systematic review and meta-analysis protocol.
Water, sanitation, and hygiene interventions often fail to show long-term impact on diarrhoeal and/or intestinal parasite risk in many low- and middle-income countries. Less attention has been paid to wider contextual factors that may contribute to high levels of contamination in the domestic environment such as household flooring. The purpose of this study will be to assess the association between diarrhoeal and/or intestinal parasite infection status and unimproved/unfinished flooring in low- and middle-income countries. We will conduct a comprehensive search of published studies (randomized controlled trials, non-randomized controlled trials, and observational studies) that examined the association between unimproved/unfinished household flooring and diarrhoeal and/or intestinal parasite infection status from January 1, 1980, onwards with no language restriction. The primary outcome will include diarrhoeal and/or intestinal parasite infection status. Databases to be searched include EMBASE, MEDLINE, Web of Science, and Google Scholar. The secondary outcome will be the association between specific pathogens (laboratory confirmed) and unimproved/unfinished household flooring. Independent screening for eligible studies using defined criteria and data extraction will be completed in duplicate and independently. Any discrepancies between the two reviewers will be resolved by consensus and/or arbitration by a third researcher. If data permits, random effects models will be used where appropriate. Subgroup and additional analyses will be conducted to explore the potential sources of heterogeneity (e.g. age group, geographical region) and potential risk of bias of included studies. This review will provide a comprehensive examination of a possible association between suboptimal household flooring and increased risk of enteric pathogen infection, highlight gaps for future research in high risk areas, and inform intervention design for future planned studies in Kenya and/or elsewhere in the region. PROSPERO registration number: CRD42019156437.


Evaluation of potential risks associated with recombination in transgenic plants expressing viral sequences.
Virus-resistant transgenic plants have been created primarily through the expression of viral sequences. It has been hypothesized that recombination between the viral transgene mRNA and the RNA of an infecting virus could generate novel viruses. As mRNA/viral RNA recombination can occur in virus-resistant transgenic plants, the key to testing this risk hypothesis is to compare the populations of recombinant viruses generated in transgenic and non-transgenic plants. This has been done with two cucumoviral systems, involving either two strains of cucumber mosaic virus (CMV), or CMV and the related tomato aspermy virus (TAV). Although the distribution of the sites of recombination in the CMV/CMV and TAV/CMV systems was quite different, equivalent populations of recombinant viruses were observed in both cases. These results constitute the first comparison of the populations of recombinants in transgenic and non-transgenic plants, and suggest that there is little risk of emergence of recombinant viruses in these plants, other than those that could emerge in non-transgenic plants.


Canonical wnt signaling in dendritic cells regulates Th1/Th17 responses and suppresses autoimmune neuroinflammation.
Breakdown in immunological tolerance to self-Ags or uncontrolled inflammation results in autoimmune disorders. Dendritic cells (DCs) play an important role in regulating the balance between inflammatory and regulatory responses in the periphery. However, factors in the tissue microenvironment and the signaling networks critical for programming DCs to control chronic inflammation and promote tolerance are unknown. In this study, we show that wnt ligand-mediated activation of β-catenin signaling in DCs is critical for promoting tolerance and limiting neuroinflammation. DC-specific deletion of key upstream (lipoprotein receptor-related protein [LRP]5/6) or downstream (β-catenin) mediators of canonical wnt signaling in mice exacerbated experimental autoimmune encephalomyelitis pathology. Mechanistically, loss of LRP5/6-β-catenin-mediated signaling in DCs led to an increased Th1/Th17 cell differentiation but reduced regulatory T cell response. This was due to increased production of proinflammatory cytokines and decreased production of anti-inflammatory cytokines such as IL-10 and IL-27 by DCs lacking LRP5/6-β-catenin signaling. Consistent with these findings, pharmacological activation of canonical wnt/β-catenin signaling delayed experimental autoimmune encephalomyelitis onset and diminished CNS pathology. Thus, the activation of canonical wnt signaling in DCs limits effector T cell responses and represents a potential therapeutic approach to control autoimmune neuroinflammation.


Melissa officinalis L. - A review of its traditional uses, phytochemistry and pharmacology.
Melissa officinalis L. is a medicinal plant that has long been used in different ethno-medical systems especially in the European Traditional Medicine and the Iranian Traditional Medicine for the treatment of several diseases. It is also widely used as a vegetable and to add flavor to dishes This review aimed to provide a summary on the botanical characterization, traditional uses, phytochemistry, pharmacological activities, pharmacokinetics and toxicity of M. officinalis, and discusses research gaps and future opportunities for investigations on this plant. We extensively reviewed major unpublished old texts, and published and electronic literature on traditional medicines of different regions of the world to find traditional uses of M. officinalis. Electronic databases including Web of Science, PubMed, ScienceDirect, Google Scholar and Scopus were searched to find articles (published between 1956 and 2015) on pharmacology and phytochemistry of M. officinalis. Traditional uses of M. officinalis have been recorded mostly in European countries, Mediterranean region and Middle East countries. Phytochemical investigations revealed that this plant contains volatile compounds, triterpenoids, phenolic acids and flavonoids. Crude extracts and pure compounds isolated from M. officinalis exhibited numerous pharmacological effects, from which only anxiolytic, antiviral and antispasmodic activities of this plant as well as its effects on mood, cognition and memory have been shown in clinical trials. AChE inhibitory activity, stimulation of the acetylcholine and GABAA receptors, as well as inhibition of matrix metallo proteinase-2 are the main mechanisms proposed for the widely discussed neurological effects of this plant. Modern pharmacological studies have now validated many traditional uses of M. officinalis. The data reviewed here revealed that M. officinalis is a potential source for the treatment of a wide range of diseases especially anxiety and some other CNS disorders, though confirmatory trials are warranted to substantiate these effects in the clinical setting. Data regarding many aspects of this plant such as mechanisms of actions, pharmacokinetics, adverse effects of the extracts, potential interactions with standard-of-care medications and active compounds is still limited which call for additional studies particularly in humans.


Cryo-EM structure of a helicase loading intermediate containing ORC-Cdc6-Cdt1-MCM2-7 bound to DNA.
In eukaryotes, the Cdt1-bound replicative helicase core MCM2-7 is loaded onto DNA by the ORC-Cdc6 ATPase to form a prereplicative complex (pre-RC) with an MCM2-7 double hexamer encircling DNA. Using purified components in the presence of ATP-γS, we have captured in vitro an intermediate in pre-RC assembly that contains a complex between the ORC-Cdc6 and Cdt1-MCM2-7 heteroheptamers called the OCCM. Cryo-EM studies of this 14-subunit complex reveal that the two separate heptameric complexes are engaged extensively, with the ORC-Cdc6 N-terminal AAA+ domains latching onto the C-terminal AAA+ motor domains of the MCM2-7 hexamer. The conformation of ORC-Cdc6 undergoes a concerted change into a right-handed spiral with helical symmetry that is identical to that of the DNA double helix. The resulting ORC-Cdc6 helicase loader shows a notable structural similarity to the replication factor C clamp loader, suggesting a conserved mechanism of action.


Health, biodiversity, and natural resource use on the Amazon frontier: an ecosystem approach.
This study aims to improve the health of rural Amazonian communities through the development and application of a participatory ecosystem approach to human health assessment. In the study area marked seasonal fluctuations dictate food availability, water quality and disease outbreak. Determining the causal linkages between ecosystem variables, resource use and health required a variety of forms of inquiry at multiple scales with local participation. Landscape spatial mapping of resource use demonstrated the diversity of the ecological resources upon which communities depend. Household surveys detailed family and individual consumption and production patterns. Anthropometric measurements, parasite loading, water quality and anemia levels were used as indicators of health status. This was complemented with an ethnographic and participatory health assessment that provided the foundation for developing community action plans addressing health issues. Discussion is focused on three attributes of an ecosystem approach; (a) methodological pluralism, (b) cross-scale interactions and (c) participatory action research.


Effects of short-term varenicline administration on emotional and cognitive processing in healthy, non-smoking adults: a randomized, double-blind, study.
Varenicline is an effective and increasingly prescribed drug for smoking cessation, but has been associated with depressive symptoms and suicidal behavior. However, it remains unclear whether those changes in mood and behavior are directly related to varenicline use, or caused by smoking cessation itself or reflects depression and suicidality rates in smokers, independent of treatment. To investigate the influence of varenicline on mood and behavior independent of smoking and smoking cessation, we assessed the effects of varenicline on emotional processing (a biomarker of depressogenic effects), emotion-potentiated startle reactivity, impulsivity (linked with suicidal behavior), and cognitive performance in non-smoking subjects. We used a randomized, double-blind design, in which we administered varenicline or placebo to healthy subjects over 7 days (0.5 mg/day first 3 days, then 1 mg/day). Cognitive and emotional processing was assessed by a battery of computerized tasks and recording of emotion-potentiated startle response. A total of 41 subjects were randomized, with 38 subjects included in the analysis. The varenicline group did not differ from placebo in terms of negative biases in emotional processing or mood. However, compared with placebo, the varenicline group scored higher on working and declarative memory. In conclusion, short-term varenicline use did not influence negative biases in emotional processing or impulsivity in non-smoking subjects, thereby not supporting direct depressogenic or suicidal risk behavior-inducing effects. In contrast, varenicline may have cognitive-enhancing effects.


Clinical assessment of HIV-associated lipodystrophy in an ambulatory population.
To identify clinical factors associated with prevalence of fat atrophy (lipoatrophy) and fat accumulation (lipoaccumulation) in HIV-1 infected patients. Evaluation of HIV-1 infected patients seen for routine care between 1 October and 31 December 1998 in the eight HIV Outpatient Study (HOPS) clinics. Eight clinics specializing in the care of HIV-1 infected patients. A total of 1077 patients were evaluated for signs of fat maldistribution. A standardized set of questions and specific clinical signs were assessed. Demographic, clinical and pharmacological data for each patient were also included in the analysis. Demographic, immunologic, virologic, clinical, laboratory, and drug treatment factors were assessed in stratified and multivariate analyses for their relationship to the presence and severity of fat accumulation and atrophy. Independent factors for moderate/severe lipoatrophy for 171 patients were increasing age, any use of stavudine, use of indinavir for longer than 2 years, body mass index (BMI) loss, and measures of duration and severity of HIV disease. Independent risk factors for moderate/severe fat accumulation for 104 patients were increasing age, BMI gain, measures of amount and duration of immune recovery, and duration of antiretroviral therapy (ART). The number of non-drug risk factors substantially increased the likelihood of lipoatrophy. If non-drug risk factors were absent, lipoatrophy was unusual regardless of the duration of drug use. HIV-associated lipodystrophy is associated with several host, disease, and drug factors. While prevalence of lipoatrophy increased with the use of stavudine and indinavir, and lipoaccumulation was associated with duration of ART, other non-drug factors were strongly associated with both fat atrophy and accumulation.


Physiological response of the European hedgehog to predator and nonpredator odour.
A respiratory chamber was used to investigate physiological responses of hedgehogs to predator and nonpredator odour cues, introduced by passing air through different faecal suspensions. Five recently caught hedgehogs showed a significant increase in oxygen consumption (29% +/- 18% at 95% c.i.) when treated with badger (predator) faecal suspension, and 10 hedgehogs held in captivity for ca. 2 years showed no significant response. The responses of recently caught hedgehogs to odour from badger (predator) and roe deer (non-predator) faecal suspensions were then investigated, over a range of concentrations spanning 3 orders of magnitude. Five hedgehogs were tested with badger odour; of these, 3 were also tested with roe deer odour. The mean rise in oxygen consumption was significantly greater in response to badger than to roe deer faecal suspension, but there was no significant variation in strength of response over the range of concentrations tested, and increased oxygen consumption was not associated with any increase in levels of visible activity. The responses are interpreted as an increase in arousal in response to a potential predatory threat. Lack of response in the captive-held animals raises questions about the welfare of such animals following release. Energy costs and foraging inefficiency associated with arousal may be significant factors of foraging decisions involving predation risk, but more naturalistic measurements would be required for any quantitative analysis.


Randomized trial comparing traditional Chinese medical acupuncture, therapeutic massage, and self-care education for chronic low back pain.
Because the value of popular forms of alternative care for chronic back pain remains uncertain, we compared the effectiveness of acupuncture, therapeutic massage, and self-care education for persistent back pain. We randomized 262 patients aged 20 to 70 years who had persistent back pain to receive Traditional Chinese Medical acupuncture (n = 94), therapeutic massage (n = 78), or self-care educational materials (n = 90). Up to 10 massage or acupuncture visits were permitted over 10 weeks. Symptoms (0-10 scale) and dysfunction (0-23 scale) were assessed by telephone interviewers masked to treatment group. Follow-up was available for 95% of patients after 4, 10, and 52 weeks, and none withdrew for adverse effects. Treatment groups were compared after adjustment for prerandomization covariates using an intent-to-treat analysis. At 10 weeks, massage was superior to self-care on the symptom scale (3.41 vs 4.71, respectively; P =.01) and the disability scale (5.88 vs 8.92, respectively; P<.001). Massage was also superior to acupuncture on the disability scale (5.89 vs 8.25, respectively; P =.01). After 1 year, massage was not better than self-care but was better than acupuncture (symptom scale: 3.08 vs 4.74, respectively; P =.002; dysfunction scale: 6.29 vs 8.21, respectively; P =.05). The massage group used the least medications (P<.05) and had the lowest costs of subsequent care. Therapeutic massage was effective for persistent low back pain, apparently providing long-lasting benefits. Traditional Chinese Medical acupuncture was relatively ineffective. Massage might be an effective alternative to conventional medical care for persistent back pain.


Can gout mimic a soft tissue tumour?
Gout is a disease where an abnormal collection of uric crystals (gouty tophi) can often be found in the foot and ankle. Occasionally such tophies are found at unusual sites and this can pose a challenge to the clinician where diagnosis and treatment is concerned. We describe a patient who presented to our oncology department with a swelling on his right thigh which was clinically diagnosed to be a soft tissue sarcoma, but after further investigations it turned out to be a gouty tophi. The purpose of this case report is to emphasise the variable presentation of gout and the challenges that can be faced by clinicians in diagnosing a soft tissue swelling.


Metamorphosis of cycloalkenes for the divergent total synthesis of polycyclic indole alkaloids.
From tryptamine and secologanine, nature generates monoterpene indole alkaloids with an unprecedented level of skeletal diversity through a 'couple-divert' sequence. Intrigued by this biosynthetic machinery, new strategies and tactics to access skeletally distinct natural products are continuously emerging. In this Tutorial Review, we'll present a simplified view of nature's logic for biosynthesis and the representative strategies and tactics developed for the divergent total synthesis of monoterpene indole alkaloids. Our group has been developing a 'couple-divert' approach with the strategic use of cycloalkene as a pluripotent motif in the synthetic design and has developed an integrated oxidation/reduction/cyclization (iORC) sequence for transforming functionalized cycloalkenes, easily available by a variety of cross-coupling reactions, to skeletally diverse natural products. The integration of controlled regio-, chemo- and stereo-selective cyclization and heteroannulation reactions into these domino sequences will be highlighted.