The calcium-sensing receptor modulates the prostaglandin E2 pathway in intestinal inflammation.
Introduction: The prostaglandin E2 (PGE2) pathway is one of the main mediators of intestinal inflammation. As activation of the calcium-sensing receptor (CaSR) induces expression of inflammatory markers in the colon, we assessed the impact of the CaSR on the PGE2 pathway regulation in colon cancer cells and the colon in vitro and in vivo. Methods and Results: We treated CaSR-transfected HT29 and Caco-2 colon cancer cell lines with different orthosteric ligands or modulators of the CaSR and measured gene expression and PGE2 levels. In CaSR-transfected HT29CaSR-GFP and Caco-2CaSR-GFP cells, the orthosteric CaSR ligand spermine and the positive allosteric CaSR modulator NPS R-568 both induced an inflammatory state as measured by IL-8 gene expression and significantly increased the expression of the PGE2 pathway key enzymes cyclooxygenase (COX)-2 and/or prostaglandin E2 synthase 1 (PGES-1). Inhibition of the CaSR with the calcilytic NPS 2143 abolished the spermine- and NPS R-568-induced pro-inflammatory response. Interestingly, we observed cell-line specific responses as e.g. PGES-1 expression was affected only in HT29CaSR-GFP but not in Caco-2CaSR-GFP cells. Other genes involved in the PGE2 pathway (COX-1, or the PGE2 receptors) were not responsive to the treatment. None of the studied genes were affected by any CaSR agonist in GFP-only transfected HT29GFP and Caco-2GFP cells, indicating that the observed gene-inducing effects of spermine and R-568 were indeed mediated by the CaSR. In vivo, we had previously determined that treatment with the clinically approved calcimimetic cinacalcet worsened symptoms in a dextran sulfate sodium (DSS)-induced colitis mouse model. In the colons of these mice, cinacalcet significantly induced gene expression of PGES-2 and the EP3 receptor, but not COX-2; while NPS 2143 increased the expression of the PGE2-degrading enzyme 15-hydroxyprostaglandin dehydrogenase (15-PGDH). Importantly, neither treatment had any effect on the colons of non-DSS treated mice. Discussion: Overall, we show that activation of the CaSR induces the PGE2 pathway, albeit with differing effects in vitro and in vivo. This may be due to the different microenvironment in vivo compared to in vitro, specifically the presence of a CaSR-responsive immune system. Since calcilytics inhibit ligand-mediated CaSR signaling, they may be considered for novel therapies against inflammatory bowel disease.


Healthcare seeking and treatment for menopausal symptoms in the United States.
A population-based study was used to describe healthcare seeking behavior for menopausal symptoms and treatment among women 40-65 years old in the United States. Participants were recruited into the Menopause Epidemiology Study from the KnowledgePanel(SM), which is selected by random digit dialing and probability sampling from the US population. From this source, 6201 women 40-65 years old were contacted and 4402 women participated. From the 3135 peri- and postmenopausal women, detailed information was obtained on menopausal symptoms, healthcare seeking, medication usage, and symptom relief from the medication. Many women (60%) reported seeking health care for their menopausal symptoms. More than half of these women sought health care in the past 12 months. Vasomotor symptoms were the most frequently reported menopause symptoms across all races/ethnicities, and the most common symptoms discussed with a health care professional. One-third of the women (34%) used only hormone therapies, 12% used complementary and/or alternative medicines, and 16% used both for treatment of menopausal symptoms. This study has shown that a large number of women consult healthcare providers for menopausal symptoms, indicating these symptoms are bothersome. Yet, in the United States, there is considerable variation in the symptomatology, healthcare seeking, and use of therapies for menopausal symptoms across cultures. To alleviate these symptoms women have tried alternative treatments as well as hormone therapies, yet many women did not get complete relief of specific symptoms.


Evaluation of partnerships in a transnational family violence prevention network using an integrated knowledge translation and exchange model: a mixed methods study.
Family violence is a significant and complex public health problem that demands collaboration between researchers, practitioners, and policymakers for systemic, sustainable solutions. An integrated knowledge translation network was developed to support joint research production and application in the area. The purpose of this study was to determine the extent to which the international Preventing Violence Across the Lifespan (PreVAiL) Research Network built effective partnerships among its members, with a focus on the knowledge user partner perspective. This mixed-methods study employed a combination of questionnaire and semi-structured interviews to understand partnerships two years after PreVAiL's inception. The questionnaire examined communication, collaborative research, dissemination of research, research findings, negotiation, partnership enhancement, information needs, rapport, and commitment. The interviews elicited feedback about partners' experiences with being part of the network. Five main findings were highlighted: i) knowledge user partner involvement varied across activities, ranging from 11% to 79% participation rates; ii) partners and researchers generally converged on their assessment of communication indicators; iii) partners valued the network at both an individual level and to fulfill their organizations' mandates; iv) being part of PreVAiL allowed partners to readily contact researchers, and partners felt comfortable acting as an intermediary between PreVAiL and the rest of their own organization; v) application of research was just emerging; partners needed more actionable insights to determine ways to move forward given the research at that point in time. Our results demonstrate the importance of developing and nurturing strong partnerships for integrated knowledge translation. Our findings are applicable to other network-oriented partnerships where a diversity of stakeholders work to address complex, multi-faceted public health problems.


Effects of body region and time on hair cortisol concentrations in chimpanzees (Pan troglodytes).
Hair cortisol concentrations (HCC) are increasingly recognized as an integrated measure of the systemic cortisol secretion. Yet, we still know very little about confounding effects on HCC in animals. The present study therefore used hair from semi-wild and zoo living chimpanzees to investigate (1) intra-individual variability of HCC (body-region effect), and (2) the stability of HCC along the hair shaft (traditionally called the washout effect). Our results indicate that absolute HCC varied substantially between certain body regions, but a factor analysis revealed that these HCC differences were mainly attributable to one common source of variance. Thus, hair from all body regions provides similar biological signals and can be mixed, albeit at the cost of a lower signal-to-noise ratio. With regard to potential underlying mechanisms, we studied skin blood flow, as observed through thermal images from one chimpanzee. We found the general HCC pattern was reflected in differences in surface body temperature observed in this individual in three out of four body regions. In a separate set of samples, we found first evidence to suggest that the systematic cortisol decrease along the hair shaft, as observed in humans, is also present in chimpanzee hair. The effect was more pronounced in semi-wild than in zoo chimpanzees presumably due to more exposure to ambient weather conditions.


Neuron activity related to short-term memory.
Nerve cells in the monkey's prefrontal cortex and nucleus medialis dorsalis of the thalamus show changes of firing frequency associated with the performance of a delayed response test. Most cells increase firing during the cue presentation period or at the beginning of the ensuing delay; spike discharge highler than that in intertrial periods is present in some cells throughout the delay. These changes are interpreted as suggestive evidence of a role of frontothalamic circuits in the attentive process involved in short-term memory


Overexpressed DNA polymerase iota regulated by JNK/c-Jun contributes to hypermutagenesis in bladder cancer.
Human DNA polymerase iota (pol ι) possesses high error-prone DNA replication features and performs translesion DNA synthesis. It may be specialized and strictly regulated in normal mammalian cells. Dysregulation of pol ι may contribute to the acquisition of a mutator phenotype. However, there are few reports describing the transcription regulatory mechanism of pol ι, and there is controversy regarding its role in carcinogenesis. In this study, we performed the deletion and point-mutation experiment, EMSA, ChIP, RNA interference and western blot assay to prove that c-Jun activated by c-Jun N-terminal kinase (JNK) regulates the transcription of pol ι in normal and cancer cells. Xeroderma pigmentosum group C protein (XPC) and ataxia-telangiectasia mutated related protein (ATR) promote early JNK activation in response to DNA damage and consequently enhance the expression of pol ι, indicating that the novel role of JNK signal pathway is involved in DNA damage response. Furthermore, associated with elevated c-Jun activity, the overexpression of pol ι is positively correlated with the clinical tumor grade in 97 bladder cancer samples and may contribute to the hypermutagenesis. The overexpressed pol ι-involved mutagenesis is dependent on JNK/c-Jun pathway in bladder cancer cells identifying by the special mutation spectra. Our results support the conclusion that dysregulation of pol ι by JNK/c-Jun is involved in carcinogenesis and offer a novel understanding of the role of pol ι or c-Jun in mutagenesis.


Evaluation of the relationship between cranial magnetic resonance imaging findings and clinical status in children with cerebral palsy
The objective of this study was to evaluate the relationship between cranial magnetic resonance imaging (MRI) findings and clinical features in cerebral palsy (CP). Children aged 3 to 18 years, who were followed with the diagnosis of CP between January 2012 and September 2015, were included. The type of CP was classified using the European Cerebral Palsy Monitoring Group's classification system and then, patients were divided into two groups as spastic or nonspastic groups. The Gross Motor Function Classification System (GMFCS) was used to determine the level of mobility. According to the GMFCS, levels 1, 2, and 3 were grouped as mobile, and levels 4 and 5 were grouped as immobile. Cranial MRI findings were reevaluated by a voluntarily radiologist and grouped as periventricular leukomalacia (PVL) (grades 1, 2, and 3), cerebral atrophy, migration anomaly, cerebellar involvement, basal ganglion involvement, and normal MRI findings. Sixty-two patients were enrolled. The rate of mobile patients did not differ between the spastic and nonspastic groups. The incidence of PVL was significantly higher in cases of prematurity and spastic CP (p < 0.05). The rate of mobilization was significantly lower and the rate of epilepsy was significantly higher in patients with PVL. Immobile patients were more common among cases of grade 3 PVL (p < 0.05). The most common cranial MRI pathology was PVL, and the presence of PVL and its grade might help clinically assess the patient's CP type and level of mobilization. While pathology was observed mostly in cranial MRI in cases of CP with similar clinical features, the fact that cranial MRI was completely normal for 14.5% of the cases suggests that there may be some pathologies that we could not identify with today's imaging technology.


Rice inhibits intestinal secretions.
Rice-based oral rehydration solutions have been found to be effective in decreasing stool output and improving intestinal absorption in acute infectious diarrhea. Recent tests of the ability of a rice extract to inhibit intestinal secretions were undertaken, and it was shown that an isolated fraction of rice extract decreased intestinal losses by actively inhibiting the chloride channel.


Rapid depletion of budding yeast proteins by fusion to a heat-inducible degron.
One effective way to study the biological function of a protein in vivo is to inactivate it and see what happens to the cell. For proteins that are dispensable for cell viability, the corresponding gene can simply be deleted from its chromosomal locus. The study of essential proteins is more challenging, however, because the function of the protein must be inactivated conditionally. Here, we describe a method that allows the target protein to be depleted rapidly and conditionally, so that the immediate effects on the cell can be examined. The chromosomal locus of a budding yeast gene is modified so that a "heat-inducible degron cassette" is added to the N terminus of the encoded protein, causing it to be degraded by a specific ubiquitin-mediated pathway when cells are shifted from 24 degrees to 37 degrees C. Degradation requires recognition of the degron cassette by the evolutionarily conserved Ubr1 protein, which is associated with a ubiquitin-conjugating enzyme. To promote rapid and conditional depletion of the target protein, we use a yeast strain in which expression of the UBR1 gene can be either repressed or strongly induced. Degron strains are constructed by a simple "one-step" approach using the polymerase chain reaction.


Generation of the membrane attack complex during haemodialysis: impact of classical and alternative pathway components.
1. The generation of the plasma SC5b-9 complex (the soluble form of the membrane attack complex) during haemodialysis was studied in 29 patients together with markers of the classical (C4d fragment) and alternative (Bb fragment) pathways and the common iC3b fragment. 2. In the patients dialysed with Hemophan and Cuprophan membranes, a rapid increase in the plasma levels of SC5b-9 complex, Bb fragment and iC3b fragment occurred within 15 min of the initiation of the haemodialysis procedure. The SC5b-9 complex and the Bb fragment concentrations remained at a plateau until 120 min thereafter. The iC3b fragment level showed a continual decline. 3. The C4d fragment concentrations remained unchanged, indicating that generation of the membrane attack complex is most likely due only to activation of the alternative pathway. 4. The generation of the membrane attack complex occurred throughout the full haemodialysis period. In patients treated with Polysulfon membranes no statistically significant variation in these components was noted. 5. The study shows that the membrane attack complex may be generated during the full period of the haemodialysis session and is a stable index of biocompatibility. 6. Moreover, it seems be involved directly in haemodialysis-associated phenomena.


Role of alternative oxidase gene in pathogenesis of Cryptococcus neoformans.
We identified a homologue of the alternative oxidase gene in a screen to identify genes that are preferentially transcribed in response to a shift to 37 degrees C in the human-pathogenic yeast Cryptococcus neoformans. Alternative oxidases are nucleus-encoded mitochondrial proteins that have two putative roles: they can function in parallel with the classic cytochrome oxidative pathway to produce ATP, and they may counter oxidative stress within the mitochondria. The C. neoformans alternative oxidase gene (AOX1) was found to exist as a single copy in the genome, and it encodes a putative protein of 401 amino acids. An aox1 mutant strain was created using targeted gene disruption, and the mutant strain was reconstituted to wild type using a full-length AOX1. Compared to both the wild-type and reconstituted strains, the aox1 mutant strain was not temperature sensitive but did have significant impairment of both respiration and growth when treated with inhibitors of the classic cytochrome oxidative pathway. The aox1 mutant strain was also found to be more sensitive to the oxidative stressor tert-butyl hydroperoxide. The aox1 mutant strain was significantly less virulent than both the wild type and the reconstituted strain in the murine inhalational model, and it also had significantly impaired growth within a macrophage-like cell line. These data demonstrate that the alternative oxidase of C. neoformans can make a significant contribution to metabolism, has a role in the yeast's defense against exogenous oxidative stress, and contributes to the virulence composite of this organism, possibly by improving survival within phagocytic cells.


Reduced granulation tissue and wound strength in the absence of α11β1 integrin.
Previous wound healing studies have failed to define a role for either α1β1 or α2β1 integrin in fibroblast-mediated wound contraction, suggesting the involvement of another collagen receptor in this process. Our previous work demonstrated that the integrin subunit α11 is highly induced during wound healing both at the mRNA and protein level, prompting us to investigate and dissect the role of the integrin α11β1 during this process. Therefore, we used mice with a global ablation of either α2 or α11 or both integrin subunits and investigated the repair of excisional wounds. Analyses of wounds demonstrated that α11β1 deficiency results in reduced granulation tissue formation and impaired wound contraction, independently of the presence of α2β1. Our combined in vivo and in vitro data further demonstrate that dermal fibroblasts lacking α11β1 are unable to efficiently convert to myofibroblasts, resulting in scar tissue with compromised tensile strength. Moreover, we suggest that the reduced stability of the scar is a consequence of poor collagen remodeling in α11(-/-) wounds associated with defective transforming growth factor-β-dependent JNK signaling.


The unique burden of rare cancer caregiving: caregivers of patients with Erdheim-Chester disease.
Research examining the experience of informal caregivers (ICs) for patients with rare cancers is limited. This was a mixed-methods pilot study of 14 ICs for patients with Erdheim-Chester disease (ECD), an ultra-rare neoplasm. Participants were predominantly female and over half provided at least 60% of their loved one's care. Participants completed measures of the impact of caregiving, caregiver burden, unmet needs, quality of life, anxiety, and depression. Participants reported substantial impact of caregiving, including limiting (50%) or discontinuing (21%) paid employment, and exhausting financial savings (43%). ICs reported a moderate level of burden with five (38%) reporting risk for burnout. While participants reported anxiety (64%) and depression (14%), their overall quality of life was favorable. Semi-structured interviews highlighted factors related to the distress and isolation of navigating a rare cancer. ECD ICs report burden and distress shaped by the experience of providing care for a patient with a rare cancer.


The role of interparticle and external forces in nanoparticle assembly.
The past 20 years have witnessed simultaneous multidisciplinary explosions in experimental techniques for synthesizing new materials, measuring and manipulating nanoscale structures, understanding biological processes at the nanoscale, and carrying out large-scale computations of many-atom and complex macromolecular systems. These advances have led to the new disciplines of nanoscience and nanoengineering. For reasons that are discussed here, most nanoparticles do not 'self-assemble' into their thermodynamically lowest energy state, and require an input of energy or external forces to 'direct' them into particular structures or assemblies. We discuss why and how a combination of self- and directed-assembly processes, involving interparticle and externally applied forces, can be applied to produce desired nanostructured materials.


Laser induced explosive vapor and cavitation resulting in effective irrigation of the root canal. Part 2: evaluation of the efficacy.
Limited information exists regarding the efficacy of laser activated irrigation (LAI) on removal of root canal debris. This study compares the efficacy of LAI for removal of debris in root canals as compared to conventional irrigation (CI) and passive ultrasonic irrigation (PUI). A splitted tooth model was constructed with straight roots prepared to a 0.06 taper and an apical diameter of ISO 40. A vertical groove was cut in the canal wall at 2-6 mm to the end of the canal in one halve of the root canal wall and filled with dentinal debris. In group 1 root canals were irrigated with 2.5% NaOCl by hand (20 seconds) with the needle 1 mm short from the apical stop, in group 2 NaOCl was ultrasonically activated (20 seconds) with an Irrisafe tip 1 mm short from the apical stop, and in group 3 NaOCl was activated with an Er,Cr:YSGG laser (Z2 Endolase tip -200 microm fiber, four times for 5 seconds, 75 mJ, 20 Hz, stationary at 5 mm from the apical stop). The remaining quantity of dentin debris in the groove was evaluated using a scoring system. LAI resulted in significantly less debris than PUI (P<0.005) and CI (P<0.0005). PUI also showed significantly less debris than CI (P<0.005). Under the conditions of this study LAI is statistically significantly more effective in removing artificially placed dentin debris in a root canal as PUI and CI.


Discovery of the gene signature for acute lung injury in patients with sepsis.
The acute respiratory distress syndrome (ARDS)/acute lung injury (ALI) was described 30 yr ago, yet making a definitive diagnosis remains difficult. The identification of biomarkers obtained from peripheral blood could provide additional noninvasive means for diagnosis. To identify gene expression profiles that may be used to classify patients with ALI, 13 patients with ALI + sepsis and 20 patients with sepsis alone were recruited from the Medical Intensive Care Unit of the University of Pittsburgh Medical Center, and microarrays were performed on peripheral blood samples. Several classification algorithms were used to develop a gene signature for ALI from gene expression profiles. This signature was validated in an independently obtained set of patients with ALI + sepsis (n = 8) and sepsis alone (n = 1). An eight-gene expression profile was found to be associated with ALI. Internal validation found that the gene signature was able to distinguish patients with ALI + sepsis from patients with sepsis alone with 100% accuracy, corresponding to a sensitivity of 100%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 100%. In the independently obtained external validation set, the gene signature was able to distinguish patients with ALI + sepsis from patients with sepsis alone with 88.9% accuracy. The use of classification models to develop a gene signature from gene expression profiles provides a novel and accurate approach for classifying patients with ALI.


Source retrieval is not properly differentiated from object retrieval in early schizophrenia: an fMRI study using virtual reality.
Source memory, the ability to identify the context in which a memory occurred, is impaired in schizophrenia and has been related to clinical symptoms such as hallucinations. The neurobiological underpinnings of this deficit are not well understood. Twenty-five patients with recent onset schizophrenia (within the first 4.5 years of treatment) and twenty-four healthy controls completed a source memory task. Participants navigated through a 3D virtual city, and had 20 encounters of an object with a person at a place. Functional magnetic resonance imaging was performed during a subsequent forced-choice recognition test. Two objects were presented and participants were asked to either identify which object was seen (new vs. old object recognition), or identify which of the two old objects was associated with either the person or the place being presented (source memory recognition). Source memory was examined by contrasting person or place with object. Both patients and controls demonstrated significant neural activity to source memory relative to object memory, though activity in controls was much more widespread. Group differences were observed in several regions, including the medial parietal and cingulate cortex, lateral frontal lobes and right superior temporal gyrus. Patients with schizophrenia did not differentiate between source and object memory in these regions. Positive correlations with hallucination proneness were observed in the left frontal and right middle temporal cortices and cerebellum. Patients with schizophrenia have a deficit in the neural circuits which facilitate source memory, which may underlie both the deficits in this domain and be related to auditory hallucinations.


Penicillium arizonense, a new, genome sequenced fungal species, reveals a high chemical diversity in secreted metabolites.
A new soil-borne species belonging to the Penicillium section Canescentia is described, Penicillium arizonense sp. nov. (type strain CBS 141311T = IBT 12289T). The genome was sequenced and assembled into 33.7 Mb containing 12,502 predicted genes. A phylogenetic assessment based on marker genes confirmed the grouping of P. arizonense within section Canescentia. Compared to related species, P. arizonense proved to encode a high number of proteins involved in carbohydrate metabolism, in particular hemicellulases. Mining the genome for genes involved in secondary metabolite biosynthesis resulted in the identification of 62 putative biosynthetic gene clusters. Extracts of P. arizonense were analysed for secondary metabolites and austalides, pyripyropenes, tryptoquivalines, fumagillin, pseurotin A, curvulinic acid and xanthoepocin were detected. A comparative analysis against known pathways enabled the proposal of biosynthetic gene clusters in P. arizonense responsible for the synthesis of all detected compounds except curvulinic acid. The capacity to produce biomass degrading enzymes and the identification of a high chemical diversity in secreted bioactive secondary metabolites, offers a broad range of potential industrial applications for the new species P. arizonense. The description and availability of the genome sequence of P. arizonense, further provides the basis for biotechnological exploitation of this species.


Is low-level respiratory resistive loading during exercise perceived as breathlessness?
1. The effect of adding low-level (2.7 cmH2O 1(-1) s) external respiratory resistive loads on exercise-induced breathlessness has been examined in naive normal subjects; the intensity of this loading was chosen to simulate that confronting an asthmatic subject during exercise. 2. Each of 18 subjects performed two separate tests in which workload was oscillated while the respiratory loading was changed every minute between no loading, inspiratory loading only, and inspiratory plus expiratory loading. Each loading condition was given three times, and both these changes and those in workload were unpredictable as far as the subject was concerned. 3. The purpose was to 'confuse' subjects and obtain subjective estimates of their intensity of breathlessness independent of any expectation associated solely with the readily perceptible changes in external resistances to breathing. The study design was balanced for the group as a whole, both in terms of workload and respiratory loading condition. 4. The addition of these respiratory resistive loads during exercise did not result in a significant increase in the intensity of breathlessness. 5. Estimates of the rate of work of breathing revealed that this increased more with respiratory loading than it did as ventilation rose throughout the test; on the other hand, the intensity of breathlessness increased by a greater extent with continued exercise compared with the changes accompanying the addition of respiratory loads. 6. It is concluded that the intensity of the sensation of breathlessness experienced by normal subjects during exercise is not simple a reflection of an increased rate of work of breathing being performed by the respiratory muscles.(ABSTRACT TRUNCATED AT 250 WORDS)


JOURNAL CLUB: Computer-Aided Detection of Lung Nodules on CT With a Computerized Pulmonary Vessel Suppressed Function.
The purpose of this study is to evaluate radiologists' performance in detecting actionable nodules on chest CT when aided by a pulmonary vessel image-suppressed function and a computer-aided detection (CADe) system. A novel computerized pulmonary vessel image-suppressed function with a built-in CADe (VIS/CADe) system was developed to assist radiologists in interpreting thoracic CT images. Twelve radiologists participated in a comparative study without and with the VIS/CADe using 324 cases (involving 95 cancers and 83 benign nodules). The ratio of nodule-free cases to cases with nodules was 2:1 in the study. Localization ROC (LROC) methods were used for analysis. In a stand-alone test, the VIS/CADe system detected 89.5% and 82.0% of malignant nodules and all nodules no smaller than 5 mm, respectively. The false-positive rate per CT study was 0.58. For the reader study, the mean area under the LROC curve (LROCAUC) for the detection of lung cancer significantly increased from 0.633 when unaided by VIS/CADe to 0.773 when aided by VIS/CADe (p < 0.01). For the detection of all clinically actionable nodules, the mean LROC-AUC significantly increased from 0.584 when unaided by VIS/CADe to 0.692 when detection was aided by VIS/CADe (p < 0.01). Radiologists detected 80.0% of cancers with VIS/CADe versus 64.45% of cancers unaided (p < 0.01); specificity decreased from 89.9% to 84.4% (p < 0.01). Radiologist interpretation time significantly decreased by 26%. The VIS/CADe system significantly increased radiologists' detection of cancers and actionable nodules with somewhat lower specificity. With use of the VIS/CADe system, radiologists increased their interpretation speed by a factor of approximately one-fourth. Our study suggests that the technique has the potential to assist radiologists in the detection of additional actionable nodules on thoracic CT.


Missing genes, multiple ORFs, and C-to-U type RNA editing in Acrasis kona (Heterolobosea, Excavata) mitochondrial DNA.
Discoba (Excavata) is an ancient group of eukaryotes with great morphological and ecological diversity. Unlike the other major divisions of Discoba (Jakobida and Euglenozoa), little is known about the mitochondrial DNAs (mtDNAs) of Heterolobosea. We have assembled a complete mtDNA genome from the aggregating heterolobosean amoeba, Acrasis kona, which consists of a single circular highly AT-rich (83.3%) molecule of 51.5 kb. Unexpectedly, A. kona mtDNA is missing roughly 40% of the protein-coding genes and nearly half of the transfer RNAs found in the only other sequenced heterolobosean mtDNAs, those of Naegleria spp. Instead, over a quarter of A. kona mtDNA consists of novel open reading frames. Eleven of the 16 protein-coding genes missing from A. kona mtDNA were identified in its nuclear DNA and polyA RNA, and phylogenetic analyses indicate that at least 10 of these 11 putative nuclear-encoded mitochondrial (NcMt) proteins arose by direct transfer from the mitochondrion. Acrasis kona mtDNA also employs C-to-U type RNA editing, and 12 homologs of DYW-type pentatricopeptide repeat (PPR) proteins implicated in plant organellar RNA editing are found in A. kona nuclear DNA. A mapping of mitochondrial gene content onto a consensus phylogeny reveals a sporadic pattern of relative stasis and rampant gene loss in Discoba. Rampant loss occurred independently in the unique common lineage leading to Heterolobosea + Tsukubamonadida and later in the unique lineage leading to Acrasis. Meanwhile, mtDNA gene content appears to be remarkably stable in the Acrasis sister lineage leading to Naegleria and in their distant relatives Jakobida.


Biodegradable coronary scaffolds: their future and clinical and technological challenges.
Angioplasty and stenting are standard treatment options for both stabile occlusive coronary artery disease and acute myocardial infarctions. Over the last years, several biodegradable stent systems have entered pre-clinical and clinical evaluation and into clinical practice. A strong supporting scaffold is necessary after angioplasty to prevent elastic recoil of the vessel but in the long term a permanent metallic stent will only impair normal physiology of the artery wall. Thus, the main advantage of a resorbable system is the potential for better vessel recovery and function in the long term. The new stent systems differ from traditional stents in size and biological responses and questions have risen regarding their mechanical strength and increased risk of stent thrombosis. Here, we present current treatment options with biodegradable scaffolds, discuss further key areas for improvements and review novel technological advances in the context of all up-to-date clinical trial information. New material choices are also covered as well as special considerations for pre-clinical testing.


Colour polymorphism in birds: causes and functions.
We studied polymorphism in all species of birds that are presently known to show intraspecific variation in plumage colour. At least three main mechanisms have been put forward to explain the maintenance of polymorphism: apostatic, disruptive and sexual selection. All of them make partly different predictions. Our aims were to investigate evolutionary causes and adaptive functions of colour polymorphism by taking into account a number of ecological and morphological features of polymorphic species. Overall, we found 334 species showing colour polymorphism, which is 3.5% of all bird species. The occurrence of colour polymorphism was very high in Strigiformes, Ciconiiformes, Cuculiformes and Galliformes. Phylogenetically corrected analysis using independent contrasts revealed that colour polymorphism was maximally expressed in species showing a daily activity rhythm extended to day/night, living in both open and closed habitats. All these findings support the hypothesis that colour polymorphism probably evolved under selective pressures linked to bird detectability as affected by variable light conditions during activity period. Thus, we conclude that selective agents may be prey, predators and competitors, and that colour polymorphism in birds may be maintained by disruptive selection.


Leukaemia in young children living in the vicinity of German nuclear power plants.
A case control study was conducted where cases were children younger than 5 years (diseased between 1980 and 2003) registered at the german childhood cancer registry (GCCR). Population-based matched controls (1:3) were selected from the corresponding registrar's office. Residential proximity to the nearest nuclear power plant was determined for each subject individually (with a precision of about 25 m). The report is focused on leukaemia and mainly on cases in the inner 5-km zone around the plants. The study includes 593 leukaemia cases and 1,766 matched controls. All leukaemia combined show a statistically significant trend for 1/distance with a positive regression coefficient of 1.75 [lower 95%-confidence limit (CL): 0.65]; for acute lymphoid leukaemia 1.63 (lower 95%-CL: 0.39), for acute nonlymphocytic leukaemia 1.99 (lower 95%-CL: -0.41). This indicates a negative trend for distance. Cases live closer to nuclear power plants than the randomly selected controls. A categorical analysis shows a statistically significant odds ratio of 2.19 (lower 95%-CL: 1.51) for residential proximity within 5 km compared to residence outside this area. This result is largely attributed to cases in previous studies of the GCCR (especially in the inner zone) as there is clearly some overlap between those studies. The result was not to be expected under current radiation-epidemiological knowledge. Considering that there is no evidence of relevant accidents and that possible confounders could not be identified, the observed positive distance trend remains unexplained.


New haplotypes of the Plasmodium falciparum chloroquine resistance transporter (pfcrt) gene among chloroquine-resistant parasite isolates.
Mutations in the Plasmodium falciparum chloroquine resistance transporter (pfcrt) gene were examined to assess their associations with chloroquine resistance in clinical samples from Armopa (Papua) and Papua New Guinea. In Papua, two of the five pfcrt haplotypes found were new: SVIET from Armopa and CVIKT from an isolate in Timika. There was also a strong association (P < 0.0001) between the pfcrt 76T allele and chloroquine resistance in 50 samples. In Papua New Guinea, mutations in the pfcrt gene were observed in 15 isolates with chloroquine minimum inhibitory concentrations (MICs) of 16-64 pmol, while the remaining six isolates, which had a wild-type pfcrt gene at codon 76, had MICs of 2-8 pmol. These observations confirm that mutations at codon 76 in the pfcrt gene are present in both in vivo and in vitro cases of chloroquine resistance, and that detection of the pfcrt 76T allele could predict potential chloroquine treatment failures.


Waveform capnography: an alternative to physician gestalt in determining optimal intubating conditions after administration of paralytic agents.
We sought to evaluate the utility of waveform capnography (WC) in detecting paralysis, by using apnoea as a surrogate determinant, as compared with clinical gestalt during rapid sequence intubation. Additionally, we sought to determine if this improves the time to intubation and first pass success rates through more consistent and expedient means of detecting optimal intubating conditions (ie, paralysis). A prospective observational cohort study of consecutively enrolled patients was conducted from April to June 2016 at an academic, urban, level 1 trauma centre in New York City. Nasal cannula WC was used to determine the presence of apnoea as a surrogate measure of paralysis versus physician gestalt (ie, blink test, mandible relaxation, and so on). One hundred patients were enrolled (50 in the WC group and 50 in the gestalt group). There were higher proportions of failure to determine optimal intubating conditions (ie, paralysis) in the gestalt group (32%, n=16) versus the WC group (6%, n=3), absolute difference 26, 95% CI 10 to 40. Time to intubation was longer in the gestalt group versus the WC group (136 seconds vs 116 seconds, absolute difference 20 seconds 95% CI 14 to 26). First pass success rates were higher in the WC group verses the gestalt group (92%, 95% CI 85 to 97 vs 88%, 95% CI 88 to 95, absolute difference 4%, 95% CI 1 to 8). These preliminary results demonstrate WC may be a useful objective measure to determine the presence of paralysis and optimal in tubating conditions in RSI.


Identification and treatment of eating disorders in the primary care setting.
Eating disorders, which are associated with a host of adverse medical morbidities, negative psychological sequelae, and considerable reductions in quality of life, should be diagnosed and treated promptly. However, primary care physicians may find it uniquely challenging to detect eating disorders in their early stages, before obvious physical problems arise and while psychological symptoms are subtle. Although psychological symptoms may dominate the presentation, the physician is an integral member of the treatment team and is in a unique role to diagnose and treat eating disorders. This clinical review surveys the eating disorders literature, identified by searching MEDLINE and PubMed for articles published from January 1, 1983, to September 30, 2009, using the following keywords: anorexia nervosa, bulimia nervosa, eating disorders, eating disorders NOS, binge eating, binge eating disorder, and night eating syndrome. This review also focuses on practical issues faced by primary care physicians in the management of these conditions and other issues central to the care of these complex patients with medical and psychiatric comorbid conditions.


Dietary patterns derived with multiple methods from food diaries and breast cancer risk in the UK Dietary Cohort Consortium.
In spite of several studies relating dietary patterns to breast cancer risk, evidence so far remains inconsistent. This study aimed to investigate associations of dietary patterns derived with three different methods with breast cancer risk. The Mediterranean Diet Score (MDS), principal components analyses (PCA) and reduced rank regression (RRR) were used to derive dietary patterns in a case-control study of 610 breast cancer cases and 1891 matched controls within four UK cohort studies. Dietary intakes were collected prospectively using 4- to 7-day food diaries and resulting food consumption data were grouped into 42 food groups. Conditional logistic regression models were used to estimate odds ratios (ORs) for associations between pattern scores and breast cancer risk adjusting for relevant covariates. A separate model was fitted for post-menopausal women only. The MDS was not associated with breast cancer risk (OR comparing first tertile with third 1.20 (95% CI 0.92; 1.56)), nor the first PCA-derived dietary pattern, explaining 2.7% of variation of diet and characterized by cheese, crisps and savoury snacks, legumes, nuts and seeds (OR 1.18 (95% CI 0.91; 1.53)). The first RRR-derived pattern, a 'high-alcohol' pattern, was associated with a higher risk of breast cancer (OR 1.27; 95% CI 1.00; 1.62), which was most pronounced in post-menopausal women (OR 1.46 (95% CI 1.08; 1.98)). A 'high-alcohol' dietary pattern derived with RRR was associated with an increased breast cancer risk; no evidence of associations of other dietary patterns with breast cancer risk was observed in this study.


Political or dental power in private and public service provision: a study of municipal expenditures for child dental care.
Both professionals and politicians may affect expenditures for highly professional services provided in the public and private sector. We investigated Danish publicly financed child dental care with a special focus on the influence of politicians and dentists on the expenditure level. By studying spatial patterns in expenditure levels across municipalities, we are able to test the influences of these two main actors and the networks through which learning is achieved. Four hypotheses on the existence of different spatial spillover effects are tested. The empirical analysis is based on annual data from 1996 to 2001 for 226 Danish municipalities, thus allowing for the control for heterogeneity between municipalities and for intra-municipal correlations across time. The results point to differences in expenditures between municipalities with privately and publicly produced dental care. Furthermore, dentists appear to be the most important actors for the spatial spillover effects, and these effects are especially strong for municipalities situated in the same county that use private dental clinics. There is no evidence of political spatial spillover effects between municipalities.


Nodular lymphocyte predominant hodgkin lymphoma and T cell/histiocyte rich large B cell lymphoma--endpoints of a spectrum of one disease?
In contrast to the commonly indolent clinical behavior of nodular lymphocyte predominant Hodgkin lymphoma (NLPHL), T cell/histiocyte rich large B cell lymphoma (THRLBCL) is frequently diagnosed in advanced clinical stages and has a poor prognosis. Besides the different clinical presentations of these lymphoma entities, there are variants of NLPHL with considerable histopathologic overlap compared to THRLBCL. Especially THRLBCL-like NLPHL, a diffuse form of NLPHL, often presents a histopathologic pattern similar to THRLBCL, suggesting a close relationship between both lymphoma entities. To corroborate this hypothesis, we performed gene expression profiling of microdissected tumor cells of NLPHL, THRLBCL-like NLPHL and THRLBCL. In unsupervised analyses, the lymphomas did not cluster according to their entity. Moreover, even in supervised analyses, very few consistently differentially expressed transcripts were found, and for these genes the extent of differential expression was only moderate. Hence, there are no clear and consistent differences in the gene expression of the tumor cells of NLPHL, THRLBCL-like NLPHL and THRLBCL. Based on the gene expression studies, we identified BAT3/BAG6, HIGD1A, and FAT10/UBD as immunohistochemical markers expressed in the tumor cells of all three lymphomas. Characterization of the tumor microenvironment for infiltrating T cells and histiocytes revealed significant differences in the cellular composition between typical NLPHL and THRLBCL cases. However, THRLBCL-like NLPHL presented a histopathologic pattern more related to THRLBCL than NLPHL. In conclusion, NLPHL and THRLBCL may represent a spectrum of the same disease. The different clinical behavior of these lymphomas may be strongly influenced by differences in the lymphoma microenvironment, possibly related to the immune status of the patient at the timepoint of diagnosis.


Neuroprotection from retinal ischemia/reperfusion injury by NOX2 NADPH oxidase deletion.
The aim of this study was to determine whether NOX2, one of the homologs of NADPH oxidase, plays a role in neuronal cell death during retinal ischemia. Ischemia reperfusion (I/R) injury was generated in C57/BL6 and NOX2(-/-) mice by increasing the intraocular pressure (IOP) to 110 mm Hg for 40 minutes followed by reperfusion. Quantitative PCR and Western blot analysis were performed to measure NOX2 expression. Reactive oxygen species (ROS) formation was assessed by dihydroethidium imaging of superoxide formation and Western blot analysis for tyrosine nitration. TUNEL assay was performed to determine cell death at 3 days after I/R. Survival of neurons within the ganglion cell layer (GCL) was assessed at 7 days after I/R by confocal morphometric imaging of retinal wholemounts immunostained with NeuN antibody. Activation of mitogen-activated protein kinases and nuclear factor κB (NF-κΒ) was measured by Western blot analysis. NOX2 mRNA and protein and ROS were significantly increased in wild-type I/R retinas. This effect was associated with a 60% decrease in the number of GCL neurons and a 10-fold increase in TUNEL-positive cells compared with the fellow sham control eyes. Phosphorylation of ERK and NF-κB was significantly increased in wild-type I/R retinas. Each of these effects was markedly attenuated in the NOX2(-/-) retina (P < 0.01). These data demonstrate that the deletion of NOX2 can reduce I/R-induced cell death and preserve retinal GCL neurons after I/R injury. The neuronal cell injury caused by I/R is associated with the activation of ERK and NF-κB signaling mechanisms.


Patient and carer experience of obtaining regular prescribed medication for chronic disease in the English National Health Service: a qualitative study.
The increasing burden of chronic disease is recognised globally. Within the English National Health Service, patients with chronic disease comprise of half of all consultations in primary care, and 70% of inpatient bed days. The cost of prescribing long-term medications for those with physical chronic diseases is rising and there is a drive to reduce medicine wastage and costs. While current policies in England are focused on the latter, there has been little previous research on patient experience of ordering and obtaining regular medication for their chronic disease. This paper presents findings from England of a qualitative study and survey of patients and their carers' experiences of community and primary care based services for physical chronic diseases. Although not the primary focus of the study, the results highlighted particular issues around service delivery of repeat prescriptions. We conducted 21 qualitative in-depth interviews with 30 patients and family carers' in two Primary Care Trusts in England. Participants were receiving community based care for diabetes, respiratory, neurological or complex co-morbidities, and ranged in age from 39-92 years old. We used a broadly inductive approach to enable themes around patient experience to emerge from the data. While the study sought to gain an overview of patient experience, the findings suggested that the processes associated with ordering and obtaining regular medication - the repeat prescription, was most frequently described as a recurring hassle of managing a long-term condition. Issues for patients and carers included multiple journeys to the surgery and pharmacy, lack of synchrony and dissatisfaction with the length of prescriptions. Much literature exists around medication waste and cost, which led to encouragement from the NHS in England to reduce dosage units to a 28-day supply. While there has been an acknowledgement that longer supplies may be suitable for people with stable chronic conditions, it appears that there is limited evidence on the impact of shorter length prescriptions on patient and carer experience, adherence and health outcomes. Recent policy documents within England also fail to address possible links between patient experience, adherence and flaws within repeat prescription service delivery.


Mechanism of differential Zika and dengue virus neutralization by a public antibody lineage targeting the DIII lateral ridge.
We previously generated a panel of human monoclonal antibodies (mAbs) against Zika virus (ZIKV) and identified one, ZIKV-116, that shares germline usage with mAbs identified in multiple donors. Here we show that ZIKV-116 interferes with ZIKV infection at a post-cellular attachment step by blocking viral fusion with host membranes. ZIKV-116 recognizes the lateral ridge of envelope protein domain III, with one critical residue varying between the Asian and African strains responsible for differential binding affinity and neutralization potency (E393D). ZIKV-116 also binds to and cross-neutralizes some dengue virus serotype 1 (DENV1) strains, with genotype-dependent inhibition explained by variation in a domain II residue (R204K) that potentially modulates exposure of the distally located, partially cryptic epitope. The V-J reverted germline configuration of ZIKV-116 preferentially binds to and neutralizes an Asian ZIKV strain, suggesting that this epitope may optimally induce related B cell clonotypes. Overall, these studies provide a structural and molecular mechanism for a cross-reactive mAb that uniquely neutralizes ZIKV and DENV1.


Involvement of Gi in the inhibition of adenylate cyclase by cannabimimetic drugs.
The cellular mechanism of action of the cannabimimetic drugs is examined using cultured cells. In membranes from N18TG2 neuroblastoma cells and the neuroblastoma X glioma hybrid cells, NG108-15, the psychoactive cannabinoid drugs and their nantradol analogs could inhibit adenylate cyclase activity. This response was not observed in either the soluble adenylate cyclase from rat sperm or membrane-bound adenylate cyclases from C6 glioma or S49 lymphoma cells. This cellular selectivity provides further evidence for the existence of specific receptors for the cannabimimetic compounds. Receptor-mediated inhibition of adenylate cyclase requires the presence of a guanine nucleotide-binding protein complex, Gi. Gi can be functionally inactivated as a result of an ADP-ribosylation modification catalyzed by pertussis toxin. The present study demonstrates that pertussis toxin treatment of cells abolished the cannabimimetic response in intact cells and in membranes derived therefrom. The action of pertussis toxin required NAD+ as substrate for in vitro modification of neuroblastoma membranes. Furthermore, pertussis toxin was able to catalyze the labeling of a neuroblastoma membrane protein in vitro using [32P] NAD+ under conditions similar to those by which attenuation of the cannabimimetic inhibition of adenylate cyclase could be demonstrated. This evidence demonstrates the requirement for a functional Gi in the action of cannabimimetic drugs.


Who is responsible for your health: is it you, your doctor or the new technologies?
The aim of the paper is to disentangle the roles that patients, physicians and technology can have on patient health outcomes. The analysis focuses on patients suffering from hypercholesterolemia. Using a large and detailed dataset of patients collected by the Italian College of General Practitioners (SIMG) over the period 2001-2006, we observe the existence of heterogeneity in the time needed to reach an optimal level of health stock. We firstly explore whether patients recovering faster exhibit lower hospitalization rates. Secondly, we study the determinants of the speed of recovery to a good health status. Results suggest that a 10% increase in the speed of recovery reduces hospitalization rates by 1% in the general sample and by 1.25% in patients in primary prevention. Furthermore, we show that recovering to a good health status is a multifaceted phenomenon, with technology explaining from 54 to 68% of the total effect.


Early migration characteristics of a hydroxyapatite-coated femoral stem: an RSA study.
Measurement of early stem subsidence can be used to predict the likelihood of long-term femoral component loosening and clinical failure. Data that examines the early migration pattern of clinically proven stems will provide clinicians with useful baseline data with which to compare new stem designs. This study was performed to evaluate the early migration pattern of a hydroxyapatite-coated press-fit femoral component that has been in use for over ten years. We enrolled 30 patients who underwent THA for osteoarthritis. The median age was 70 years (range, 55-80 years). Patients were clinically assessed using the Harris hip score. Radiostereometric analysis was used to evaluate stem migration at three to four days, six months, one year and two years. We observed a mean subsidence of 0.73 mm at six months, 0.62 mm at one year and 0.58 mm at two years and a mean retroversion of 1.82° at six months, 1.90° at one year and 1.59° at two years. This data suggests that subsidence is confined to the first six months after which there was no further subsidence. The results from this study can be compared with those from novel cementless stem designs to help predict the long-term outcome one may expect from new cementless stem designs.


[Carcinosarcoma of the esophagus - a case report].
The patient was a 79-year-old male complaining of fever, loss of appetite, cough, and a feeling of obstruction when swallowing. He was diagnosed with pneumonia and admitted as an emergency case the same day. Because an esophagus space-occupying lesion was observed on chest computed tomography(CT), in addition to evidence of pneumonia, an upper gastrointestinal endoscopy was performed. A tumor, protruding into the lumen of the esophagus, was seen in the midesophagus, 25-30 cm from the incisors. Because of the narrow lumen, only a fine caliber fiber could be passed. Biopsy results indicated only necrotic tissue, and a repeat biopsy was performed, with similar histological findings. No esophagobronchial fistulas were observed during bronchoscopy. We therefore diagnosed the patient with aspiration pneumonia, secondary to esophageal narrowing by a tumor. A preoperative diagnosis of cancer could not be made, and no distant organ metastasis was detected, but surgery was indicated because of the narrowing of the esophagus, regardless of the possibility of cancer. After the pneumonia improved, total thoracic esophagectomy was performed through a right thoracolaparotomy, plus a 3- region cervico-thoraco-abdominal lymph node dissection. Pathological examination of the surgical specimen revealed autolysis of the superficial layer with progression to necrosis and associated inflammation. The majority of the tumor was composed of spindle-shaped atypical cells, but because a very small transitional area between squamous cell carcinoma and sarcoma was noted, a diagnosis of carcinosarcoma was made. Depth of invasion was sm3, and no regional lymph node metastasis was detected. The patient's disease was classified as pT1b(sm3)N0M0, StageI. No definite diagnosis was made preoperatively. Although carcinosarcoma of the esophagus is rare, the endoscopic findings are characteristic. We report this case with a review of the literature.


Treatment decision making based on the published natural history and growth rate of small meningiomas.
Definitive data allowing clinicians to predict which meningioma patients will fail to respond to conservative management are lacking. To address this need, the authors systematically reviewed the published literature regarding the natural history of small, untreated meningiomas. The authors performed a systematic review of the existing literature on untreated meningiomas that were followed with serial MR imaging. They summarize the published linear rates of tumor growth, and the risk factors for development of new or worsened symptoms during follow-up by using a stratified chi-square test. The search methods identified 22 published studies reporting on 675 patients with untreated meningiomas followed by serial MR imaging. Linear growth rates varied significantly: no growth was the most common rate, although reports of more aggressive tumors noted growth rates of up to a 93% linear increase in size per year. The authors found that few patients with initial tumor diameters < 2 cm went on to develop new or worsened symptoms over a median follow-up period of 4.6 years. Patients with initial tumor diameters of 2-2.5 cm demonstrated a marked difference in the rate of symptom progression if their tumors grew > 10% per year, compared with those tumors growing ≤ 10% per year (42% vs 0%; p < 0.001, chi-square test). Patients with tumors between > 2.5 and 3 cm in initial size went on to develop new or worsened symptoms 17% of the time. This systematic review of the literature regarding the clinical behavior of untreated meningiomas suggests that most meningiomas ≤ 2.5 cm in diameter do not proceed to cause symptoms in the approximately 5-year period following their discovery. Those that do cause symptoms can usually be predicted with close radiographic follow-up. Based on these findings, the authors suggest the importance of observation in the early course of treatment for small asymptomatic meningiomas, especially those with an initial diameter < 2 cm.


Bioassay-guided isolation of cytotoxic flavonoids from aerial parts of Coronopus didymus.
Coronopus didymus Linn. (Brassicaceae) is a medicinal plant used traditionally as antipyretic, expectorant, to purify blood and for alleviating symptoms of pain, inflammations, malaria, wounds and cancer. The present study was designed to isolate and identify the cytotoxic compounds responsible for anticancer activity from this traditionally useful medicinal plant. Bioassay-guided fractionation of the ethanolic extract of aerial parts of C. didymus allowed the isolation of compounds responsible for anticancer activity. Their structures were elucidated by UV Spectroscopy (with shift reagents), ESI-MS and NMR spectral data. Preliminary anticancer activity of ethanolic extract, different fractions and isolated compounds was assessed through MTT in vitro cytotoxicity assay in a dose dependent manner against human cancer cell lines (HeLa and LN18) and normal 293T cells. Three flavonoids namely 5,7,4'-trihydroxy-3'-methoxyflavone-4'-O-β-D-glucoside (1), 5,7,4'-trihydroxy-3'-methoxyflavone-4'-O-(6''-acetyl)-β-D-glucoside (2) and 5,7,4'-trihydroxy-3'-methoxy flavone (3), were isolated from aerial parts. Compound 1 was identified for the first time from the genus Coronopus. All the compounds 1-3 showed promising activity against HeLa cells with IC50 values of 43.50, 0.63 and 3.67 µM, respectively. Significant result was also obtained with compound 3 against LN18 cells with IC50 value of 46.63 µM. The cytotoxic activity of the crude extract and fractions which may largely be due to its major isolated constituents, flavonoids 1-3, against both HeLa and LN18 cells provides a scientific basis for the ethnopharmacological use of C. didymus as anticancer agent.


Closed-loop for type 1 diabetes - an introduction and appraisal for the generalist.
Rapid progress over the past decade has been made with the development of the 'Artificial Pancreas', also known as the closed-loop system, which emulates the feedback glucose-responsive functionality of the pancreatic beta cell. The recent FDA approval of the first hybrid closed-loop system makes the Artificial Pancreas a realistic therapeutic option for people with type 1 diabetes. In anticipation of its advent into clinical care, we provide a primer and appraisal of this novel therapeutic approach in type 1 diabetes for healthcare professionals and non-specialists in the field. Randomised clinical studies in outpatient and home settings have shown improved glycaemic outcomes, reduced risk of hypoglycaemia and positive user attitudes. User input and interaction with existing closed-loop systems, however, are still required. Therefore, management of user expectations, as well as training and support by healthcare providers are key to ensure optimal uptake, satisfaction and acceptance of the technology. An overview of closed-loop technology and its clinical implications are discussed, complemented by our extensive hands-on experience with closed-loop system use during free daily living. The introduction of the artificial pancreas into clinical practice represents a milestone towards the goal of improving the care of people with type 1 diabetes. There remains a need to understand the impact of user interaction with the technology, and its implication on current diabetes management and care.


Attitudes towards pharmacotherapy in late-life bipolar disorder.
Lithium remains a first-line treatment for bipolar disorder, but clinicians have considerable concern over potential adverse effects, especially in older adults. Older patients' attitude towards lithium has not been investigated, even though negative attitudes are closely associated with reduced adherence. We examine the attitude towards lithium pharmacotherapy in older adults with bipolar disorder. In a cross-sectional study of 78 patients aged >60 years with bipolar disorder, the association between lithium use and attitudes towards psychotropic pharmacotherapy was assessed using the Drug Attitude Inventory (DAI-10), including multivariate analyses. Compared to patients using alternative psychopharmacological treatments (n =30), lithium users (n=48) showed higher self-reported contentedness, subjective somatic health, and social functioning scores. Although 58.7% of lithium users reported severe adverse effects, lithium users had more positive attitudes towards psychotropic pharmacotherapy compared to non-users (DAI-10 mean score 6.0 vs. 3.9, p =0.01), and this effect was independent of potential confounders. Older bipolar patients using lithium have a more positive attitude towards psychotropic pharmacotherapy, despite high rates of adverse effects. Future longitudinal studies could investigate whether positive medication attitudes lead to improved treatment adherence and reduced bipolar disorder relapse in older lithium users.


The role of cell proliferation and crypt fission in adenoma aggressiveness: a comparison of ileoanal pouch and rectal adenomas in familial adenomatous polyposis.
In patients with familial adenomatous polyposis (FAP), ileoanal pouch cancer is rare whereas rectal cancer is common, despite polyp initiation at the two sites being similar at the molecular level. This study investigated whether the disparity in adenoma aggressiveness reflects underlying differences in histogenesis. Normal mucosal biopsies and 2-3 mm adenomas from patients with FAP were dissected into individual crypts. Crypt area, morphology, fission and mitoses were analysed for crypts from pouch, rectum and supra-anastomotic ileum. Immunohistochemistry of similar archival samples was performed for lysozyme, β-catenin and TP53 expression. The morphology of normal crypts was similar at each site, although crypt area differed. The area of normal pouch crypts was intermediate between rectum and ileum. The area of adenomatous crypts of rectum and pouch was similar, but the latter had increased asymmetrical fission. Crypt mitoses were proportional to area in all tissues, but crypt fission was reduced in adenomatous crypts from the rectum compared with the pouch. Pouch adenomas retained lysozyme expression as seen in normal ileum. Nuclear β-catenin accumulation was similar, but TP53 expression was increased in rectal adenomas. Diminutive polyps from rectum and pouch differ in morphology and proliferation. Aggressiveness in rectal polyps is not conferred by increased crypt proliferation, fission, or activation of the Wnt signalling pathway. Increased TP53 expression suggests other molecular mechanisms may be responsible. While crypt mitoses are proportional to crypt area, the threshold for fission may be site specific, indicating that tissue origin may influence histogenesis and thus malignant potential.


Phenoloxidase activity acts as a mosquito innate immune response against infection with Semliki Forest virus.
Several components of the mosquito immune system including the RNA interference (RNAi), JAK/STAT, Toll and IMD pathways have previously been implicated in controlling arbovirus infections. In contrast, the role of the phenoloxidase (PO) cascade in mosquito antiviral immunity is unknown. Here we show that conditioned medium from the Aedes albopictus-derived U4.4 cell line contains a functional PO cascade, which is activated by the bacterium Escherichia coli and the arbovirus Semliki Forest virus (SFV) (Togaviridae; Alphavirus). Production of recombinant SFV expressing the PO cascade inhibitor Egf1.0 blocked PO activity in U4.4 cell- conditioned medium, which resulted in enhanced spread of SFV. Infection of adult female Aedes aegypti by feeding mosquitoes a bloodmeal containing Egf1.0-expressing SFV increased virus replication and mosquito mortality. Collectively, these results suggest the PO cascade of mosquitoes plays an important role in immune defence against arboviruses.


GLUT-1 overexpression: Link between hemodynamic and metabolic factors in glomerular injury?
Mesangial matrix deposition is the hallmark of hypertensive and diabetic glomerulopathy. At similar levels of systemic hypertension, Dahl salt-sensitive but not spontaneously hypertensive rats (SHR) develop glomerular hypertension, which is accompanied by upregulation of transforming growth factor beta1 (TGF-beta1), mesangial matrix expansion, and sclerosis. GLUT-1 is ubiquitously expressed and is the predominant glucose transporter in mesangial cells. In mesangial cells in vitro, GLUT-1 overexpression increases basal glucose transport, resulting in excess fibronectin and collagen production. TGF-beta1 has been shown to upregulate GLUT-1 expression. We demonstrated that in hypertensive Dahl salt-sensitive (S) rats fed 4% NaCl (systolic blood pressure [SBP]: 236+/-9 mm Hg), but not in similarly hypertensive SHR (SBP: 230+/-10 mm Hg) or their normotensive counterparts (Dahl S fed 0.5% NaCl, SBP: 145+/-5 mm Hg; and Wistar-Kyoto, SBP: 137+/-3 mm Hg), there was an 80% upregulation of glomerular GLUT-1 protein expression (P< or =0.03). This was accompanied by a 2.7-fold upregulation of TGF-beta1 protein expression in glomeruli of DSH compared with DSN rats (P=0.02). TGF-beta1 expression was not upregulated and did not differ in the glomeruli of Wistar-Kyoto and SHR rats. As an in vitro surrogate of the in vivo hemodynamic stress imposed by glomerular hypertension, we used mechanical stretching of human and rat mesangial cells. We found that after 33 hours of stretching, mesangial cells overexpressed GLUT-1 (40%) and showed an increase in basal glucose transport of similar magnitude (both P< or =0.01), which could be blocked with an anti TGF-beta1-neutralizing antibody. These studies suggest a novel link between hemodynamic and metabolic factors that may cooperate in inducing progressive glomerular injury in conditions characterized by glomerular hypertension.


Nucleotide variation at the myrosinase-encoding locus, TGG1, and quantitative myrosinase enzyme activity variation in Arabidopsis thaliana.
The Arabidopsis thaliana TGG1 gene encodes thioglucoside glucohydrolase (myrosinase), an enzyme catalysing the hydrolysis of glucosinolate compounds. The enzyme is involved in plant defence against some insect herbivores, and is present in species of the order Capparales (Brassicales). Nucleotide variation was surveyed by sequencing c. 2.4 kb of the TGG1 locus in a sample of 28 worldwide A. thaliana accessions, and one Arabidopsis lyrata ssp. lyrata individual. Myrosinase activity was quantified for 27 of these same A. thaliana accessions, plus five additional others. Overall, estimated nucleotide diversity in A. thaliana was low compared to other published A. thaliana surveys, and the frequency distribution was skewed toward an excess of low-frequency variants. Furthermore, comparison to the outgroup species A. lyrata demonstrated that A. thaliana exhibited an excess of high-frequency derived variants relative to a neutral equilibrium model, suggesting a selective sweep. A. thaliana accessions differed significantly in total myrosinase activity, but analysis of variance detected no statistical evidence for an association between quantitative enzyme activity and alleles at the TGG1 myrosinase-encoding locus. We thus conclude that other, unsurveyed factors primarily affect the observed myrosinase activity levels in this species. The pattern of nucleotide variation was consistent with a model of positive selection but might also be compatible with a completely neutral model that takes into account the metapopulation behaviour of this highly inbreeding species which experienced a relatively recent worldwide expansion.


Wearable Technology for Global Surgical Teleproctoring.
In underserved communities around the world, inguinal hernias represent a significant burden of surgically-treatable disease. With traditional models of international surgical assistance limited to mission trips, a standardized framework to strengthen local healthcare systems is lacking. We established a surgical education model using web-based tools and wearable technology to allow for long-term proctoring and assessment in a resource-poor setting. This is a feasibility study examining wearable technology and web-based performance rating tools for long-term proctoring in an international setting. Using the Lichtenstein inguinal hernia repair as the index surgical procedure, local surgeons in Paraguay and Brazil were trained in person by visiting international expert trainers using a formal, standardized teaching protocol. Surgeries were captured in real-time using Google Glass and transmitted wirelessly to an online video stream, permitting real-time observation and proctoring by mentoring surgeon experts in remote locations around the world. A system for ongoing remote evaluation and support by experienced surgeons was established using the Lichtenstein-specific Operative Performance Rating Scale. Data were collected from 4 sequential training operations for surgeons trained in both Paraguay and Brazil. With continuous internet connectivity, live streaming of the surgeries was successful. The Operative Performance Rating Scale was immediately used after each operation. Both surgeons demonstrated proficiency at the completion of the fourth case. A sustainable model for surgical training and proctoring to empower local surgeons in resource-poor locations and "train trainers" is feasible with wearable technology and web-based communication. Capacity building by maximizing use of local resources and expertise offers a long-term solution to reducing the global burden of surgically-treatable disease.


Early brain perfusion improvement after ventriculoperitoneal shunt surgery in patients with idiopathic normal pressure hydrocephalus evaluated by 99mTc-HMPAO SPECT - preliminary report.
Idiopathic normal pressure hydrocephalus (iNPH) is a clinical syndrome that consists of the triad: gait disturbance, mental deterioration and urinary incontinence associated with normal cerebrospinal fluid pressure (CSF), without pre-existing abnormalities. The most popular treatment option is surgical implantation of a shunt. Brain perfusion increase occurring months or years after successful shunt surgery is well described in the literature. Early improvement of perfusion is not well documented. Therefore, the objective of the present study was to determine patterns of brain perfusion changes 3-6 days after the ventriculoperitoneal shunting in patients with iNPH by using 99mTc-HMPAO SPECT. Sixteen patients with iNPH (9 women, 7 men, mean age 64.1 ± 12.7 years) who underwent ventriculoperitoneal shunt surgery were included into the study group. Indications for implanting a shunt were based on clinical history, neuroimaging and CSF dynamic studies with an infusion test. Brain perfusion SPECT was performed 1-2 days before and 3-6 days after the surgical treatment. For comparison of perfusion before and after the surgery SPECT scans were assessed visually and semiquantitatively with voxel based analysis. No side effects were observed after the surgery. Brain perfusion improvement after shunting was observed in 10 patients (62.5%). Patterns of perfusion changes varied between patients, with combinations of different bilateral and lateralized brain regions involved. Perfusion increased in the whole brain (3 patients), in the right cerebral hemisphere (1 patient) or in the separate cerebral regions (6 patients): frontal, parietal, temporal, cerebellum, cingulate gyrus. Perfusion improvement was predominantly observed in the frontal lobes: right frontal (3 cases, 18.8%), left frontal (3 cases, 18.8%). Cerebral perfusion is recovered promptly after ventriculoperitoneal shunt surgery in about 60% of patients with iNPH. This improvement may be global or regional in different cerebral areas with prevalence of the frontal lobes.


Quality of life and patient-reported outcomes in endometrial cancer clinical trials: a call for action!
There is increasing recognition that quality of life (QoL) and patient-reported outcomes (PROs) are of fundamental importance and particularly relevant given the relatively high likelihood of long-term survival in most women with endometrial cancer (EC). However, there has been relatively little research focused on this topic. Our objective was to analyze our current knowledge and identify research questions to be included in the design of next clinical trials. Analyze and critically assess reported clinical trials in EC that have included QoL and PROs as primary or secondary end points. Surgery has a significant impact on physical and functional domains of QoL particularly in the first 6 months after diagnosis. Minimally invasive surgery is associated with less acute morbidity than open procedures and this persists over time. Lymphadenectomy is associated with increased incidence of lymphedema, important late effect. Adjuvant external irradiation may cause gastrointestinal and genitourinary symptoms that impact on physical functioning and which can persist over time. In contrast, vaginal brachytherapy has less toxicity and fewer late effects than external irradiation. The impact of treatment on sexuality has been poorly evaluated in EC survivors. There are few published data on QoL and PROs in patients treated with chemotherapy and the long-term impact has not been addressed. There is no evidence that palliative chemotherapy reduces symptoms and improves QoL. There are very few longitudinal studies on survivorship that is an important concern in EC survivors. Although there have been some studies addressing QoL and PROs in EC, we have identified deficiencies and gaps in our knowledge. Careful consideration of QoL and PROs end points and how to include them in clinical trials will result in a better appreciation of how treatments can impact on patients QoL and lead to conduct interventions to reduce late effects.


Timing of Advanced Airway Placement after Witnessed Out-of-Hospital Cardiac Arrest.
Background: Advanced airways (endotracheal tubes, supraglottic airways) are frequently placed by Emergency Medical Services (EMS) in patients with out-of-hospital cardiac arrest (OHCA). However, if an airway is to be placed, it is unknown whether this should occur early or late in the sequence of resuscitation events. This study evaluated the association between the timing of airway placement and the minute-to-minute probability of achieving return of spontaneous circulation (ROSC). Methods: This secondary analysis of Resuscitation Outcomes Consortium Prehospital Resuscitation using an Impedance Valve and Early versus Delayed (ROC PRIMED) study data included adult, non-traumatic, witnessed OHCA patients with airway placement by EMS before ROSC. The primary exposure variable was time from EMS arrival to advanced airway placement. The outcome was prehospital ROSC. Since resuscitations occur over time, a Cox proportional hazards model was fit to estimate the probability of ROSC as a function of the airway timing, adjusting for Utstein variables. Results: A total of 7,547 patients were included. Time to airway placement was 0-5 minutes in 12% of the cohort, >5-10 (36%), >10-15 (29%), >15-20 (14%), >20-25 (5%), >25-30 (2%), and >30 (2%). ROSC occurred in 43%. Time to airway had a statistically significant impact on ROSC. A negative association between the time to airway placement and the hazard of ROSC was observed, such that increasing intervals between EMS arrival and airway placement were associated with decreasing probabilities of ROSC, regardless of initial cardiac rhythm. Conclusions: EMS advanced airway placement has a time-dependent association with ROSC. In witnessed OHCA patients receiving advanced airways, early airway placement is associated with increased probability of ROSC.


Limitations to the spacing effect: demonstration of an inverted u-shaped relationship between interrepetition spacing and free recall.
The spacing effect refers to the finding that memory for repeated items improves when the interrepetition interval increases. To explain the spacing effect in free-recall tasks, a two-factor model has been put forward that combines mechanisms of contextual variability and study-phase retrieval (e.g., Raaijmakers, 2003; Verkoeijen, Rikers, & Schmidt, 2004). An important, yet untested, implication of this model is that free recall of repetitions should follow an inverted u-shaped relationship with interrepetition spacing. To demonstrate the suggested relationship an experiment was conducted. Participants studied a word list, consisting of items repeated at different interrepetition intervals, either under incidental or under intentional learn instructions. Subsequently, participants received a free-recall test. The results revealed an inverted u-shaped relationship between free recall and interrepetition spacing in both the incidental-learning condition and the intentional-learning condition. Moreover, for intentionally learned repetitions, the maximum free-recall performance was located at a longer interrepetition interval than for incidentally learned repetitions. These findings are interpreted in terms of the two-factor model of spacing effects in free-recall tasks.


The Cbl protooncogene product: from an enigmatic oncogene to center stage of signal transduction.
The c-cbl protooncogene was first identified as the cellular homologue of a viral oncogene v-cbl that induces pre-B lymphomas and myeloid leukemias in mice. Until recently, the biochemical basis for Cbl's transforming potential and its physiological role remained unclear. However, a convergence of biochemical studies in mammalian cells and genetic studies in C. elegans and Drosophila has now identified Cbl as a negative regulator of tyrosine kinase signaling. The N-terminal transforming region of Cbl (Cbl-N) and an adjacent RING finger domain are the elements most conserved during evolution. The Cbl-N region has now been shown to contain a novel phosphotyrosine-binding (PTB) domain that directly interacts with autophosphorylated tyrosine kinases via a D(N/D)XpY motif. A critical role of the PTB domain in Cbl function is demonstrated by the localization of a loss-of-function mutation in C. elegans Cbl homologue SLI-1 within this region. The corresponding mutation in human Cbl inactivates the PTB domain function and abrogates Cbl-mediated regulation of tyrosine kinase function. Recent studies have also identified a novel signaling pathway initiated by the interaction of mammalian Cbl proteins with the SH2 domains of Crk adaptor molecules, which results in Cbl's linkage with C3G, a guanine nucleotide exchange protein for Rap1 family of small G-proteins. Presently, Rap1 is thought to antagonize Ras function, although Rap1-specific targets have emerged recently. Thus, recent advances have firmly placed the little known protooncoprotein Cbl on the center stage of tyrosine kinase-mediated signal transduction.


Comparison of proximal and distal colonic muscle of the rabbit.
Mechanical length-tension properties and response to neurohumoral agents were compared for proximal and distal colonic muscle. Resting tension during stretch, acetylcholine-stimulated tension, and the total tension were determined. Proximal circular muscle developed a maximum total tension of 0.96 +/- 0.18 kg/cm2 (mean +/- SE) compared to 0.86 +/- 0.06 kg/cm2 for the distal colon (P greater than 0.05). Resting tension was 0.33 +/- 0.03 kg/cm2 for the proximal colon and 0.05 +/- 0.01 kg/cm2 for the distal colon at the length of optimal acetylcholine-stimulated tension (Lo) (P less than 0.01). Longitudinal muscle showed a similar difference for the proximal and distal colon. The high resting tension in the proximal colonic muscle was reduced by nitroprusside or calcium-free Krebs with EGTA. Dose-response curves to acetylcholine, histamine, phenylephrine, and isoproterenol were similar for the muscle of either part of the colon. Gastrin or cholecystokinin had no effect on the muscle. In summary, the circular or longitudinal muscles of the proximal and distal colon have different length-tension properties but only minimal differences in response to neurohumoral agents.


Inhibition of the complement activation by an adrenal androgen, dehydroepiandrosterone.
The effect of dehydroepiandrosterone (DEA), an adrenal androgen successfully used for preventing attacks in hereditary angioneurotic edema (HANE) patients was studied on the activation of classical and alternative complement pathway. The steroid inhibited both the spontaneous and immune activation of the classical complement pathway (CP), the former effect, however, was found to be more marked than the latter one. DEA exerted its inhibiting effect most probably by interfering with the internal activation of C1. Because DEA rendered HANE patients symptom free but induced only a slight increase in their serum C1-INH level, our present findings suggest that inhibition of CP activation may have a significance in the therapeutic effect of DEA and possibly of other androgens as well.


Use of docking peptides to design modular substrates with high efficiency for mitogen-activated protein kinase extracellular signal-regulated kinase.
The mitogen-activated protein kinase extracellular regulated kinase (ERK) plays a key role in the regulation of cellular proliferation. Mutations in the ERK cascade occur in 30% of malignant tumors. Thus understanding how the kinase identifies its cognate substrates as well as monitoring the activity of ERK is central to cancer research and therapeutic development. ERK binds to its protein targets, both downstream substrates and upstream activators, via a binding site distinct from the catalytic site of ERK. The substrate sequences that bind, or dock, to these sites on ERK influence the efficiency of phosphorylation. For this reason, simple peptide substrates containing only phosphorylation sequences typically possess low efficiencies for ERK. Appending short docking peptides derived from full-length protein substrates and activators of ERK to a phosphorylation sequence increased the affinity of ERK for the phosphorylation sequence by as much as 200-fold while only slightly diminishing the maximal velocity of the reaction. The efficiency of the phosphorylation reaction was increased by up to 150-fold, while the specificity of the substrate for ERK was preserved. Simple modular peptide substrates, which can be easily tailored to possess high phosphorylation efficiencies, will enhance our understanding of the regulation of ERK and provide a tool for the development of new kinase assays.


The in vivo pattern of AID targeting to immunoglobulin switch regions deduced from mutation spectra in msh2-/- ung-/- mice.
Immunoglobulin (Ig) class switching is initiated by deamination of C-->U within the immunoglobulin heavy chain locus, catalyzed by activation-induced deaminase (AID). In the absence of uracil-DNA glycosylase (UNG) and the homologue of bacterial MutS (MSH)-2 mismatch recognition protein, the resultant U:G lesions are not processed into switching events but are fixed by replication allowing sites of AID-catalyzed deamination to be identified by the resulting C-->T mutations. We find that AID targets cytosines in both donor and acceptor switch regions (S regions) with the deamination domains initiating approximately 150 nucleotides 3' of the I exon start sites and extending over several kilobases (the IgH intronic enhancer is spared). Culturing B cells with interleukin 4 or interferon gamma specifically enhanced deamination around Sgamma1 and Sgamma2a, respectively. Mutation spectra suggest that, in the absence of UNG and MSH2, AID may occasionally act at the mu switch region in an apparently processive manner, but there is no marked preference for targeting of the transcribed versus nontranscribed strand (even in areas capable of R loop formation). The data are consistent with switch recombination being triggered by transcription-associated, strand-symmetric AID-mediated deamination at both donor and acceptor S regions with cytokines directing isotype specificity by potentiating AID recruitment to the relevant acceptor S region.


[Effects of Chinese herbs for replenishing shen and strengthening qi on some indexes of neuro-endocrino-immune network in asthmatic rats].
To study changes of several related indexes of neuro-endocrino-immune (NEI) network in rat asthma model, and to observe effects of epimedium herb (EH, a Chinese herb for replenishing Shen) and milkvetch root (MR, a Chinese herb for strengthening qi) on these indexes. One hundred and twenty male healthy Brown Norway rats of clean grade were randomly divided into 11 groups. Their mRNA expression of corticoid release hormone (CRH) in hypothalamus was tested by Realtime-PCR, serum ACTH and CORT, IL-6, IL-4, IFN-gamma were detected with radioimmunoassay and enzyme-linked immunosorbent assay respectively and pathological changes of lung tissue were observed with hematoxylin and eosin stain. As compared with the normal group, mRNA expression of CRH in hypothalamic tissue, plasma ACTH and serum concentration of IFN-gamma not changed significantly, but serum level of CORT raised significantly and the pre-inflammatory factors IL-6 and IL-4 showed an increasing trend in the model rats. As compared with the model group, level of CRH mRNA expression in groups treated with low dose and moderate dose of both EH and the formula (HE and MR), IFN-gamma in group treated with moderate dose of EH and serum CORT in group treated with low dose of MR were higher, and serum levels of IL-4 and IL-6 in groups treated with high dose of MR were lower. Rats suffered from repeated asthmatic attack for three weeks had some disorders in indexes of NEI network. Chinese herbs for replenish Shen and strengthening qi could improve the function of hypothalamic-pituitary-adrenal axis and adjust the balance of Th1 and Th2 cytokines, which might be one of the mechanisms of Chinese herbs for treatment of repeated asthma attack.


Gene transfer may be preventive but not curative for a lysosomal transport disorder.
Cystinosis belongs to a growing class of lysosomal storage disorders (LSDs) caused by defective transmembrane proteins. The causative CTNS gene encodes the lysosomal cystine transporter, cystinosin. Currently the aminothiol cysteamine is the only drug available for reducing cystine storage but this treatment has non-negligible side effects and administration constraints. In this study, for the first time, we report viral vector-mediated CTNS gene transfer and evaluate the feasibility of this strategy as a complementary treatment. Initially, we transduced human CTNS(-/-) fibroblast cell lines and primary murine Ctns(-/-) hepatocyte cultures in vitro and demonstrated that gene transfer can reduce cystine storage. Because of age-related increase in cystine levels, we transduced hepatocytes from young (</=3 months of age) and older (>/=5 months of age) mice. Our in vitro data suggested that the efficiency of correction was age-dependent. We tested these observations in vivo: short-term (1 week) and long-term (4 weeks) CTNS-transduction significantly reduced hepatic cystine levels in young, but not older, Ctns(-/-) mice. Our data provide the proof-of-concept that gene transfer is feasible for correcting defective lysosomal transport, but suggest that, in the case of cystinosis, it could be preventive but not curative in some tissues.


Management of brain abscesses associated with halo fixation.
Halo orthoses are commonly used in the management of a variety of cervical spinal pathological conditions. Although placement of the cranial pins was initially believed to be safe with minimal complications, minor complications related to the cranial pins have the potential to create a setting for formation of cerebral abscesses. The risk of death due to cerebral abscesses has declined in the modern antibiotic era, but cerebral abscesses are associated with long-standing neurological morbidity that should not be considered negligible. Prevention by careful pin placement and hygiene and appropriate early management are crucial to prevent more serious complications. The authors report the case of a patient treated with a halo orthosis after incurring cervical spinal trauma who developed a cerebral abscess as a complication related to the cranial pins of the halo fixation device. They review the literature in an attempt to formulate a standardized treatment algorithm to prevent this disease process and to treat an abscess if it should form.


Towards the development of a hybrid-integrated chip interferometer for online surface profile measurements.
Non-destructive testing and online measurement of surface features are pressing demands in manufacturing. Thus optical techniques are gaining importance for characterization of complex engineering surfaces. Harnessing integrated optics for miniaturization of interferometry systems onto a silicon wafer and incorporating a compact optical probe would enable the development of a handheld sensor for embedded metrology applications. In this work, we present the progress in the development of a hybrid photonics based metrology sensor device for online surface profile measurements. The measurement principle along with test and measurement results of individual components has been presented. For non-contact measurement, a spectrally encoded lateral scanning probe based on the laser scanning microscopy has been developed to provide fast measurement with lateral resolution limited to the diffraction limit. The probe demonstrates a lateral resolution of ∼3.6 μm while high axial resolution (sub-nanometre) is inherently achieved by interferometry. Further the performance of the hybrid tuneable laser and the scanning probe was evaluated by measuring a standard step height sample of 100 nm.


Hemodynamic Forces Tune the Arrest, Adhesion, and Extravasation of Circulating Tumor Cells.
Metastatic seeding is driven by cell-intrinsic and environmental cues, yet the contribution of biomechanics is poorly known. We aim to elucidate the impact of blood flow on the arrest and the extravasation of circulating tumor cells (CTCs) in vivo. Using the zebrafish embryo, we show that arrest of CTCs occurs in vessels with favorable flow profiles where flow forces control the adhesion efficacy of CTCs to the endothelium. We biophysically identified the threshold values of flow and adhesion forces allowing successful arrest of CTCs. In addition, flow forces fine-tune tumor cell extravasation by impairing the remodeling properties of the endothelium. Importantly, we also observe endothelial remodeling at arrest sites of CTCs in mouse brain capillaries. Finally, we observed that human supratentorial brain metastases preferably develop in areas with low perfusion. These results demonstrate that hemodynamic profiles at metastatic sites regulate key steps of extravasation preceding metastatic outgrowth.


Omega 6 fatty acids for the primary prevention of cardiovascular disease.
Omega 6 plays a vital role in many physiological functions but there is controversy concerning its effect on cardiovascular disease (CVD) risk. There is conflicting evidence whether increasing or decreasing omega 6 intake results in beneficial effects. The two primary objectives of this Cochrane review were to determine the effectiveness of:1. Increasing omega 6 (Linoleic acid (LA), Gamma-linolenic acid (GLA), Dihomo-gamma-linolenic acid (DGLA), Arachidonic acid (AA), or any combination) intake in place of saturated or monounsaturated fats or carbohydrates for the primary prevention of CVD.2. Decreasing omega 6 (LA, GLA, DGLA, AA, or any combination) intake in place of carbohydrates or protein (or both) for the primary prevention of CVD. We searched the following electronic databases up to 23 September 2014: the Cochrane Central Register of Controlled Trials (CENTRAL) on the Cochrane Library (Issue 8 of 12, 2014); MEDLINE (Ovid) (1946 to September week 2, 2014); EMBASE Classic and EMBASE (Ovid) (1947 to September 2014); Web of Science Core Collection (Thomson Reuters) (1990 to September 2014); Database of Abstracts of Reviews of Effects (DARE) and Health Technology Assessment Database, and Health Economics Evaluations Database on the Cochrane Library (Issue 3 of 4, 2014). We searched trial registers and reference lists of reviews for further studies. We applied no language restrictions. Randomised controlled trials (RCTs) of interventions stating an intention to increase or decrease omega 6 fatty acids, lasting at least six months, and including healthy adults or adults at high risk of CVD. The comparison group was given no advice, no supplementation, a placebo, a control diet, or continued with their usual diet. The outcomes of interest were CVD clinical events (all-cause mortality, cardiovascular mortality, non-fatal end points) and CVD risk factors (changes in blood pressure, changes in blood lipids, occurrence of type 2 diabetes). We excluded trials involving exercise or multifactorial interventions to avoid confounding. Two review authors independently selected trials for inclusion, extracted the data, and assessed the risk of bias in the included trials. We included four RCTs (five papers) that randomised 660 participants. No ongoing trials were identified. All included trials had at least one domain with an unclear risk of bias. There were no RCTs of omega 6 intake reporting CVD clinical events. Three trials investigated the effect of increased omega 6 intake on lipid levels (total cholesterol, low density lipoprotein (LDL-cholesterol), and high density lipoprotein (HDL-cholesterol)), two trials reported triglycerides, and two trials reported blood pressure (diastolic and systolic blood pressure). Two trials, one with two relevant intervention arms, investigated the effect of decreased omega 6 intake on blood pressure parameters and lipid levels (total cholesterol, LDL-cholesterol, and HDL-cholesterol) and one trial reported triglycerides. Our analyses found no statistically significant effects of either increased or decreased omega 6 intake on CVD risk factors.Two studies were supported by funding from the UK Food Standards Agency and Medical Research Council. One study was supported by Lipid Nutrition, a commercial company in the Netherlands and the Dutch Ministry of Economic Affairs. The final study was supported by grants from the Finnish Food Research Foundation, Finnish Heart Research Foundation, Aarne and Aili Turnen Foundation, and the Research Council for Health, Academy of Finland. We found no studies examining the effects of either increased or decreased omega 6 on our primary outcome CVD clinical endpoints and insufficient evidence to show an effect of increased or decreased omega 6 intake on CVD risk factors such as blood lipids and blood pressure. Very few trials were identified with a relatively small number of participants randomised. There is a need for larger well conducted RCTs assessing cardiovascular events as well as cardiovascular risk factors.


Dynamics of Cellulose Nanocrystal Alignment during 3D Printing.
The alignment of anisotropic particles during ink deposition directly affects the microstructure and properties of materials manufactured by extrusion-based 3D printing. Although particle alignment in diluted suspensions is well described by analytical and numerical models, the dynamics of particle orientation in the highly concentrated inks typically used for printing via direct ink writing (DIW) remains poorly understood. Using cellulose nanocrystals (CNCs) as model building blocks of increasing technological relevance, we study the dynamics of particle alignment under the shear stresses applied to concentrated inks during DIW. With the help of in situ polarization rheology, we find that the time period needed for particle alignment scales inversely with the applied shear rate and directly with the particle concentration. Such dependences can be quantitatively described by a simple scaling relation and qualitatively interpreted in terms of steric and hydrodynamic interactions between particles at high shear rates and particle concentrations. Our understanding of the alignment dynamics is then utilized to estimate the effect of shear stresses on the orientation of particles during the printing process. Finally, proof-of-concept experiments show that the combination of shear and extensional flow in 3D printing nozzles of different geometries provides an effective means to tune the orientation of CNCs from fully aligned to core-shell architectures. These findings offer powerful quantitative guidelines for the digital manufacturing of composite materials with programmed particle orientations and properties.