Pellino-3 promotes endotoxin tolerance and acts as a negative regulator of TLR2 and TLR4 signaling.
Development of endotoxin tolerance in macrophages during sepsis reprograms Toll-like receptor 4 signaling to inhibit proinflammatory cytokines without suppressing anti-inflammatory and antimicrobial mediators and protects the host from excessive inflammation and tissue damage. However, endotoxin tolerance renders septic patients immunocompromised and unable to control secondary infections. Although previous studies have revealed the importance of several negative regulators of Toll-like receptor signaling in endotoxin tolerance, the role of Pellino proteins has not been addressed. The present report shows that the induction of endotoxin tolerance in vivo in mice and in vitro in human monocytes and THP-1 and MonoMac-6 macrophages increases the expression of Pellino-3. Overexpression of Pellino-3 in human embryonic kidney 293/Toll-like receptor 2 or 293/Toll-like receptor 4/myeloid differentiation factor-2 cells inhibited Toll-like receptor 2/4-mediated activation of nuclear factor-κB and induction of CXCL-8 mRNA, and Pellino-3 ablation increased these responses. Pellino-3-deficient THP-1 cells had elevated Toll-like receptor 2/4-driven tumor necrosis factor-α, interleukin-6 mRNA, and Toll-like receptor 4-driven CCL5 gene expression in response to Toll-like receptor agonists and heat-killed Escherichia coli and Staphylococcus aureus, cytokines controlled by the MyD88 and Toll-interleukin-1R domain-containing protein inducing interferon-β-mediated pathways, respectively. In addition, deficiency in Pellino-3 slightly increased phagocytosis of heat-killed bacteria. Transfected Pellino-3 inhibited nuclear factor-κB activation driven by overexpression of MyD88, TIR domain-containing adapter inducing interferon-β, interleukin-1R-associated kinase-1, and tumor necrosis factor receptor activator of nuclear factor-κB-binding kinase-1, TGF-β-activated kinase 1, and tumor necrosis factor receptor-associated factor-6, and inhibited interleukin-1R-associated kinase 1 modifications and tumor necrosis factor receptor activator of nuclear factor-κB-binding kinase 1 phosphorylation. Finally, Pellino-3 ablation in THP-1 decreased the extent of endotoxin tolerization. Thus, Pellino-3 is involved in endotoxin tolerance and functions as a negative regulator of Toll-like receptor 2/4 signaling.


Influence of architecture of β-tricalcium phosphate scaffolds on biological performance in repairing segmental bone defects.
Although three-dimensional (3D) β-tricalcium phosphate (β-TCP) scaffolds serve as promising bone graft substitutes for the segmental bone defect treatment, no consensus has been achieved regarding their optimal 3D architecture. In this study, we has systematically compared four types of β-TCP bone graft substitutes with different 3D architectures, including two types of porous scaffolds, one type of tubular scaffolds and one type of solid scaffolds, for their efficacy in treating segmental bone defect in a rabbit model. Our study has demonstrated that when compared to the traditional porous and solid scaffolds, tubular scaffolds promoted significantly higher amount of new bone formation in the defect regions as shown by X-ray, micro CT examinations and histological analysis, restored much greater mechanical properties of the damaged bone evidenced by the biomechanical testing, and eventually achieved the complete union of segmental defect. Moreover, the implantation of tubular scaffolds enhanced the neo-vascularization at the defect region with higher bone metabolic activities than others, as indicated by the bone scintigraphy assay. This study has further the current knowledge regarding the profound influence of overall 3D architecture of β-TCP scaffolds on their in vivo defect healing performance and illuminated the promising potential use of tubular scaffolds as effective bone graft substitute in treating large segmental bone defects.


Mechanical characterization of porcine liver properties for computational simulation of indentation on cancerous tissue.
An accurate characterization of soft biological tissue properties is essential for a realistic simulation of surgical procedures. Unconfined uniaxial compression tests with specimens affixed to the fixtures are often performed to characterize the stress-stretch curves of soft biological tissues, with which the material parameters can be obtained. However, the constrained boundary condition causes non-uniform deformation during the uniaxial test, posing challenges for accurate measurement of tissue deformation. In this study, we measured the deformation locally at the middle of liver specimens and obtained the corresponding stress-stretch curves. Since the effect of the constrained boundary condition on the local deformation of specimen is minimized, the stress-stretch curves are thus more realistic. Subsequently, we fitted the experimental stress-stretch curves with several constitutive models and found that the first-order Ogden hyperelastic material model was most suitable for characterizing the mechanical properties of porcine liver tissues. To further verify the characterized material properties, we carried out indentation tests on porcine liver specimens and compared the experimental data with computational results by using finite element simulations. A good agreement was achieved. Finally, we constructed computational models of liver tissue with a tumor and investigated the effect of the tumor on the mechanical response of the tissue under indentation. The computational results revealed that the liver specimen with tumor shows a stiffer response if the distance between the tumor and the indenter is small.


Diagnostic Yield of Lateral Decubitus Digital Subtraction Myelogram Stratified by Brain MRI Findings.
To assess the diagnostic yield of lateral decubitus digital subtraction myelography (LDDSM) and stratify LDDSM diagnostic yield by the Bern spontaneous intracranial hypotension (SIH) score of preprocedure brain MRI. This retrospective diagnostic study included consecutive adult patients investigated for SIH who underwent LDDSM. Patients without preprocedure brain and spine MRI and patients with extradural fluid collection on spine MRI (type 1 leak) were excluded. LDDSM images and brain MRIs were assessed by 2 independent blinded readers; a third reader adjudicated any discrepancies. Diagnostic yield of LDDSM was assessed, both overall and stratified by Bern SIH scoring. Of the 62 patients included in this study, 33 (53.2%) had a CSF leak identified on LDDSM. Right-sided leaks were more common (70.6%), and the most commonly identified levels of leaks were at T6, T7, and T10. No leak was found in any of the 9 patients with Bern SIH score of 2 or less. Of the 11 patients with Bern SIH score of 3-4, 5 (45.5%) had a CSF leak identified; of the 42 patients with Bern SIH score of 5 or higher, 28 (66.7%) had a CSF leak identified. LDDSM has a high diagnostic yield for finding the exact location of spinal CSF leak, and the diagnostic yield increases with higher Bern SIH score. No leaks were found in patients with Bern SIH score of 2 or less, suggesting that foregoing invasive testing such as LDDSM in these patients may be appropriate unless accompanied by high clinical suspicion. This study provides Class II evidence that for patients with suspected SIH, higher Bern SIH scores are associated with a greater likelihood of LDDSM-identified CSF leaks.


Institutionalising ELSA in the moment of breakdown?
This article discusses outcomes of a dialogue conference on 'The road ahead for ELSA in Norway: Issues of quality, influence and network cooperation' held in Oslo in December 2012. Norwegian researchers in the field of ethical, legal and social aspects of technologies (ELSA) were invited to discuss conceptual and strategic issues, as well as the setup of a researcher network. In the article I take an institutionalist approach and discuss challenges in institutionalising an ELSA network at a time when a designated ELSA funding programme is coming to an end. The research question is how the Norwegian ELSA network can succeed as a persistent network in times of greater uncertainties. The article claims that the network needs to gain legitimacy, outlines different dimensions of legitimacy and interprets the conference discussions in light of these dimensions. Central challenges and success factors facing the ELSA network are discussed and the article concludes with reflections on the potential future of ELSA in Norway. Although the article has a Norwegian context, the discussions in the article are likely to be relevant for researchers all across Europe, as similar developments are taking place also elsewhere in the European research funding context.


Enhanced mid-infrared chemical imaging (IRCI) detection of DNA microarrays.
We report on the optimization of a recently proposed mid-infrared chemical imaging (IRCI) detection method for the analysis of DNA microarrays. The improved protocol allowed for a ten-fold reduction in the time needed to generate a mosaic image of an entire microarray and the production of IR images with high contrast that would facilitate data analysis and interpretation. Advantages of using this protocol were evaluated by applying it to the analysis of four virulence genes in the genomes of 19 strains of the food bacterial pathogen Yersinia enterocolitica.


Abnormal error processing in depressive states: a translational examination in humans and rats.
Depression has been associated with poor performance following errors, but the clinical implications, response to treatment and neurobiological mechanisms of this post-error behavioral adjustment abnormality remain unclear. To fill this gap in knowledge, we tested depressed patients in a partial hospital setting before and after treatment (cognitive behavior therapy combined with medication) using a flanker task. To evaluate the translational relevance of this metric in rodents, we performed a secondary analysis on existing data from rats tested in the 5-choice serial reaction time task after treatment with corticotropin-releasing factor (CRF), a stress peptide that produces depressive-like signs in rodent models relevant to depression. In addition, to examine the effect of treatment on post-error behavior in rodents, we examined a second cohort of rodents treated with JDTic, a kappa-opioid receptor antagonist that produces antidepressant-like effects in laboratory animals. In depressed patients, baseline post-error accuracy was lower than post-correct accuracy, and, as expected, post-error accuracy improved with treatment. Moreover, baseline post-error accuracy predicted attentional control and rumination (but not depressive symptoms) after treatment. In rats, CRF significantly degraded post-error accuracy, but not post-correct accuracy, and this effect was attenuated by JDTic. Our findings demonstrate deficits in post-error accuracy in depressed patients, as well as a rodent model relevant to depression. These deficits respond to intervention in both species. Although post-error behavior predicted treatment-related changes in attentional control and rumination, a relationship to depressive symptoms remains to be demonstrated.


Therapeutic effects of sequential chemoradiotherapy with pemetrexed and cisplatin on locally advanced laryngeal cancer.
To explore the therapeutic effects of sequential chemoradiotherapy with pemetrexed and cisplatin on locally advanced laryngeal cancer (LALC). Fifty LALC patients who were treated in our hospital between January 2010 and January 2012 were selected and randomly divided into an observation group and a control group (n=25). The two groups were given conventional radiotherapy in the same manner, before which two cycles of chemotherapy were performed. The observation group intravenously infused with 500 mg/m2 pemetrexed on d1 and 25 mg/m2 cisplatin on d1-3, with 28 days as a cycle. The control group was intravenously infused with 25 mg/m2 cisplatin on d1-3 and 400 mg/m2 fluorouracil, with 28 days as a cycle. The short-term effects and adverse reactions of both groups were observed after treatment, and their survival was observed by follow-up for five years. The response rate was 84% (21/25) in the observation group and 64% (16/25) in the control group, between which the difference was statistically significant (P<0.05). The differences in the incidence rates of short-term adverse reactions such as grade III-IV gastrointestinal reactions and bone marrow suppression were not statistically significant between PC regimen (pemetrexed combined with cisplatin) and PF regimen (cisplatin combined with fluorouracil) (P>0.05). The incidence of long-term adverse reactions such as grade III-IV laryngeal edemas, laryngeal cartilage inflammation and laryngeal cartilage necrosis showed no significant differences between the two groups (P>0.05). The median survival was 3.3 years after PC chemotherapy and 2.8 years after PF chemotherapy, between which the difference was not statistically significant (P>0.05). The levels of serum tumor markers significantly decreased after PC and PF treatments compared with those before (P<0.05). Combining PC chemotherapy with radiotherapy has satisfactory short-term therapeutic effects on LALC, and the resulting adverse effects can be tolerated. Therefore, this strategy is worthy of promotion and application in clinical practice.


The openEHR Java reference implementation project.
The openEHR foundation has developed an innovative design for interoperable and future-proof Electronic Health Record (EHR) systems based on a dual model approach with a stable reference information model complemented by archetypes for specific clinical purposes.A team from Sweden has implemented all the stable specifications in the Java programming language and donated the source code to the openEHR foundation. It was adopted as the openEHR Java Reference Implementation in March 2005 and released under open source licenses. This encourages early EHR implementation projects around the world and a number of groups have already started to use this code. The early Java implementation experience has also led to the publication of the openEHR Java Implementation Technology Specification. A number of design changes to the specifications and important minor corrections have been directly initiated by the implementation project over the last two years. The Java Implementation has been important for the validation and improvement of the openEHR design specifications and provides building blocks for future EHR systems.


Lipid lowering agents, cognitive decline, and dementia: the three-city study.
The aim of this prospective cohort study was to evaluate the effects of lipid lowering agent (LLA) intake on cognitive function in 6,830 community-dwelling elderly persons. Cognitive performance (global cognitive functioning, visual memory, verbal fluency, psychomotor speed, and executive function), clinical diagnosis of dementia, and fibrate and statin use, were evaluated at baseline, and 2, 4, and 7 year follow-up. Multivariate Cox models were stratified by gender and adjusted for sociodemographic characteristics, mental and physical health including vascular risk factors, and genetic vulnerability (apolipoprotein E and cholesteryl ester transfer protein). For women but not men, fibrate use was specifically associated with an increased risk over 7 years of decline in visual memory only (HR = 1.29, 95% CI = 1.09-1.54, p = 0.004), and did not increase risk for incident dementia. This association was independent of genetic vulnerability related to apolipoprotein E and cholesteryl exchange transfer protein polymorphisms and occurred only in women with higher low density lipoprotein (LDL)-cholesterol levels and treated with fibrate (HR = 1.39, 95% CI = 1.08-1.79, p = 0.01) and not in those with lower LDL-cholesterol levels irrespective of fibrate treatment. For both genders, no significant associations were found between statins (irrespective of their lipophilicity) and either cognitive decline or dementia incidence. This prospective study, adjusting for multiple confounders, found no evidence that LLA given in late life reduced the risk of cognitive decline and dementia, but did raise the possibility that women with treatment-resistant high LDL-cholesterol may be at increased risk of decline in visual memory.


Dynamic multi-path WDM routing in a monolithically integrated 8 × 8 cross-connect.
We demonstrate for the first time WDM multi-path routing through a monolithically integrated InP/InGaAsP 8 × 8 space and wavelength selective cross-connect. Data channels are dynamically routed from four input ports to the same output port with excellent OSNR from 27.0 to 31.1 dB. Representative data paths are evaluated in terms of optical power penalty. Data routing experiments are performed using round-robin scheduling with nanosecond time-scale switching times.


Pharmacogenomics and gemcitabine.
Approximately half of lung cancer patients present with metastases, and a large proportion will develop recurrent disease, with median survival to cisplatin-based chemotherapy of 11 months. No predictive factor of response to cisplatin-based chemotherapy is yet available in clinical practice. The nucleotide excision repair system plays a major role in repairing a variety of distorting lesions, notably platinum-induced DNA adducts. ERCC1 is a leading gene in repairing cisplatin DNA damage. We carried out three different studies examining individually the role of ERCC1, RRM1, and then both, mRNA expression in paraffin-embedded pretreatment bronchial biopsies from gemcitabine/cisplatin-treated advanced non-small-cell lung cancer (NSCLC) patients. Median survival was significantly prolonged in patients with low levels of ERCC1 or RRM1. BRCA1 is involved in homologous recombination repair, and we observed that low levels of BRCA1 mRNA significantly increased survival in gemcitabine/cisplatin-treated patients. Our observations lead us to recommend that tumors be regularly assessed for ERCC1 and BRCA1 mRNA expression in order to customize gemcitabine/cisplatin treatment.


Comparison of Content on the American Board of Internal Medicine Maintenance of Certification Examination With Conditions Seen in Practice by General Internists.
Success on the internal medicine (IM) examination is a central requirement of the American Board of Internal Medicine's (ABIM's) Maintenance of Certification program (MOC). Therefore, it is important to understand the degree to which this examination reflects conditions seen in practice, one dimension of content validity, which focuses on the match between content in the discipline and the topics on the examination questions. To assess whether the frequency of questions on IM-MOC examinations were concordant with the frequency of conditions seen in practice. The 2010-2013 IM-MOC examinations were used to calculate the percentage of questions for 186 medical condition categories from the examination blueprint, which balances examination content by considering importance and frequency of conditions seen in practice. Nationally representative estimates of conditions seen in practice by general internists were estimated from the primary diagnosis for 13 832 office visits (2010-2013 National Ambulatory Medical Care Surveys) and 108 472 hospital stays (2010 National Hospital Discharge Survey). Prevalence of conditions included on the IM-MOC examination questions. The outcome measure was the concordance between the percentages of IM-MOC examination questions and the percentages of conditions seen in practice during either office visits or hospital stays for each of 186 condition categories (eg, diabetes mellitus, ischemic heart disease, liver disease). The concordance thresholds were 0.5 SD of the weighted mean percentages of the applicable 186 conditions seen in practice (0.74% for office visits; 0.51% for hospital stays). If the absolute differences between the percentages of examination questions and the percentages of office visit conditions or hospital stay conditions seen were less than the applicable concordance threshold, then the condition category was judged to be concordant. During the 2010-2013 IM-MOC examination periods, 3600 questions (180 questions per examination form) were administered and 3461 questions (96.1%) were mapped into the 186 study conditions (mean, 18.6 questions per condition). Comparison of the percentages of 186 categories of medical conditions seen in 13 832 office visits and 108 472 hospital stays with the percentages of 3461 questions on IM-MOC examinations revealed that 2389 examination questions (69.0%; 95% CI, 67.5%-70.6% involving 158 conditions) were categorized as concordant. For concordance between questions and office visits only, 2010 questions (58.08%; 95% CI, 56.43%-59.72% of all examination questions) involving 145 conditions were categorized as concordant. For concordance between questions and hospital stays only, 1456 questions (42.07%; 95% CI, 40.42%-43.71% of all examination questions) involving 122 conditions were categorized as concordant. Among questions on IM-MOC examinations from 2010-2013, 69% were concordant with conditions seen in general internal medicine practices, although some areas of discordance were identified.


Risk and prognosis of secondary bladder cancer after post-operative radiotherapy for gynecological cancer.
The aim of this study was to investigate the impacts of radiation therapy (RT) on the occurrence risk of secondary bladder cancer (SBC) and on the patients' survival outcome after being diagnosed with gynecological cancer (EC). The data was obtained from the SEER database between 1973 and 2015. Chi-squared test was used to compare the clinicopathological characteristics among the different groups. Fine and Gray's competing risk model was used to assess the cumulative incidence and occurrence risk of SBC in GC survivors. Kaplan-Meier method was utilized for survival analysis. A total of 123,476 GC patients were included, among which 31,847 (25.8%) patients received RT while 91629 (74.2%) patients did not. The cumulative incidence of SBC was 1.59% or 0.73% among patients who had received prior GC specific RT or not, respectively. All EBRT (standardized incidence ratio (SIR) =2.49, 95% CI [2.17-2.86]), brachytherapy (SIR =1.96, 95% CI [1.60-2.38]), and combinational RT modality groups (SIR =2.73, 95% CI [2.24-3.28]) had dramatically higher SBC incidence as compared to the US general population. Receiving EBRT (HR = 2.83, 95% CI [2.34-3.43]), brachytherapy (HR = 2.17, 95% CI [1.67-2.82]), and combinational RT modality (HR = 2.97, 95% CI [2.34-3.77]) were independent risk factors for SBC development. Survival detriment was observed in SBC patients who received RT after GC diagnosis, as compared to those who did not receive RT. In conclusion, patients who underwent RT after GC had an increased risk of developing bladder as a secondary primary cancer. A long-term surveillance for SBC occurrence is necessary for GC patients who have received prior RT.


Bonobos and chimpanzees exhibit human-like framing effects.
Humans exhibit framing effects when making choices, appraising decisions involving losses differently from those involving gains. To directly test for the evolutionary origin of this bias, we examined decision-making in humans' closest living relatives: bonobos (Pan paniscus) and chimpanzees (Pan troglodytes). We presented the largest sample of non-humans to date (n = 40) with a simple task requiring minimal experience. Apes made choices between a 'framed' option that provided preferred food, and an alternative option that provided a constant amount of intermediately preferred food. In the gain condition, apes experienced a positive 'gain' event in which the framed option was initially presented as one piece of food but sometimes was augmented to two. In the loss condition, apes experienced a negative 'loss' event in which they initially saw two pieces but sometimes received only one. Both conditions provided equal pay-offs, but apes chose the framed option more often in the positive 'gain' frame. Moreover, male apes were more susceptible to framing than were females. These results suggest that some human economic biases are shared through common descent with other apes and highlight the importance of comparative work in understanding the origins of individual differences in human choice.


Intakes and adequacy of potentially important nutrients for cognitive development among 5-year-old children in the Seychelles Child Development and Nutrition Study.
To assess the nutritional adequacy of Seychellois children in relation to nutrients reported to be important for cognitive development. Dietary intakes were assessed by 4 d weighed food diaries and analysed using dietary analysis software (WISP version 3·0; Tinuviel Software, UK). Individual nutrient intakes were adjusted to usual intakes and, in order to investigate adequacy, were compared with the UK Estimated Average Requirements for children aged 4-6 years. Children 5 years old were followed up as part of the Seychelles Child Development Nutrition Study (SCDNS), located in the high-fish-consuming population of Mahé, Republic of Seychelles. Analysis was carried out on a sample of 229 children (118 boys, 111 girls). Children consumed a diet of which fortified cereal and milk products contributed the most to nutrient intakes. The majority (≥80 %) of children met requirements for several nutrients important for child development including Fe, folate and Se. Adjusted dietary intakes of Cu, Zn, iodine, niacin and vitamin A were below the Estimated Average Requirement or Recommended Nutrient Intake. Mean adjusted energy intakes (boys 4769 kJ/d (1139·84 kcal/d), girls 4759 kJ/d (1137·43 kcal/d)) were lower than the estimated energy requirement (boys 5104 kJ/d (1220 kcal/d), girls 5042 kJ/d (1205 kcal/d)) for 88 % of boys and 86 % of girls. Nutrition was adequate for most children within the SCDNS cohort. Low intakes of some nutrients (including Zn, niacin and vitamin A) could reflect nutritional database inaccuracies, but may require further investigation. The study provides valuable information on the adequacy of intakes of nutrients which could affect the growth and development of Seychellois children.


Snake and bird predation drive the repeated convergent evolution of correlated life history traits and phenotype in the Izu Island Scincid lizard (Plestiodon latiscutatus).
Predation may create strong natural selection pressure on the phenotype and life history characteristics of prey species. The Izu scincid lizards (Plestiodon latiscutatus) that inhabit the four Japanese Izu Islands with only bird predators are drab brown, mature later, lay small clutches of large eggs, and hatch large neonates. In contrast, skinks on seven islands with both snake and bird predators are conspicuously colored, mature early, lay large clutches of small eggs, and hatch small neonates. We test the hypothesis that these suites of traits have evolved independently on each island via natural selection pressures from one of two predator regimes--birds-only and birds + snakes. Using two mtDNA genes and a nuclear locus, we infer a time-calibrated phylogeny of P. latiscutatus that reveals a basal split between Mikura and all islands south, and Miyake, all islands north, and the Izu Peninsula. Populations inhabiting Miyake, Niijima, Shikine, and Toshima are not monophyletic, suggesting either multiple colonizations or an artifact of incomplete lineage sorting (ILS). We therefore developed novel phylogenetic comparative analyses that assume either a multiple colonization or more restrictive single colonization ILS scenario and found 1) statistically significant support for the of different suites of phenotypic and life history characteristics with the presence of bird-only or bird + snake predator assemblages, and 2) strong phylogenetic support for at least two independent derivations of either the "bird-only" or "snakes + birds" phenotypes regardless of colonization scenario. Finally, our time-calibrated phylogeographic analysis supports the conclusion that the ancestor to modern Izu Island P. latiscutatus dispersed from the mainland to the Izu proto-islands between 3-7.6 million years ago (Ma). These lineages remained present in the area during successive formation of the islands, with one lineage re-colonizing the mainland 0.24-0.7 Ma.


Functional outcome and service engagement in major depressive disorder with psychotic features: comparisons with schizophrenia, schizoaffective disorder and bipolar disorder in a 6-year follow-up of the Cavan-Monaghan First Episode Psychosis Study (CAMFEPS).
While long-term outcome following a first psychotic episode is well studied in schizophrenia (SZ), schizoaffective disorder (SA), and bipolar disorder (BD), major depressive disorder with psychotic features (MDDP) has received less investigation. This study compares MDDP with SZ, SA, and BD at 6-year follow-up. At 6 years after a first psychotic episode, follow-up data on psychopathology, functioning, quality of life, and service engagement were obtained for 27 cases of MDDP in comparison to 60 SZ, 27 SA, and 35 BD. Positive psychotic symptoms were less prominent in MDDP and BD than in SZ and SA. Negative symptoms, impaired functioning, and reduction in objectively determined quality of life were less prominent in MDDP and BD, intermediate in SA and most prominent in SZ. However, subjectively determined quality of life was indistinguishable across diagnoses. Service engagement was highest for MDDP, intermediate for SA and BD, and lowest for SZ. At 6-year follow-up, these diagnoses are characterized by quantitative rather than qualitative differences in psychopathology, functionality, quality of life, and service engagement, with considerable overlap between them. These findings suggest that MDDP should join SZ, SA, and BD in a milieu of psychosis that transcends arbitrary boundaries.


Study on the removal efficiency of UF membranes using bacteriophages in bench-scale and semi-technical scale.
To determine the removal efficiency of ultrafiltration (UF) membranes for nano-particles in the size range of viruses the state of the art uses challenge tests with virus-spiked water. This work focuses on bench-scale and semi-technical scale experiments. Different experimental parameters influencing the removal efficiency of the tested UF membrane modules were analyzed and evaluated for bench- and semi-technical scale experiments. Organic matter in the water matrix highly influenced the removal of the tested bacteriophages MS2 and phiX174. Less membrane fouling (low ΔTMP) led to a reduced phage reduction. Increased flux positively affected phage removal in natural waters. The tested bacteriophages MS2 and phiX174 revealed different removal properties. MS2, which is widely used as a model organism to determine virus removal efficiencies of membranes, mostly showed a better removal than phiX174 for the natural water qualities tested. It seems that MS2 is possibly a less conservative surrogate for human enteric virus removal than phiX174. In bench-scale experiments log removal values (LRV) for MS2 of 2.5-6.0 and of 2.5-4.5 for phiX174 were obtained for the examined range of parameters. Phage removal obtained with differently fabricated semi-technical modules was quite variable for comparable parameter settings, indicating that module fabrication can lead to differing results. Potting temperature and module size were identified as influencing factors. In conclusion, careful attention has to be paid to the choice of experimental settings and module potting when using bench-scale or semi-technical scale experiments for UF membrane challenge tests.


Docosahexaenoic acid attenuates Western diet-induced hepatic fibrosis in Ldlr-/- mice by targeting the TGFβ-Smad3 pathway.
DHA (22:6,ω3), but not EPA (20:5,ω3), attenuates Western diet (WD)-induced hepatic fibrosis in a Ldlr(-/-) mouse model of nonalcoholic steatohepatitis. We examined the molecular basis for the differential effect of dietary EPA and DHA on WD-induced hepatic fibrosis. DHA was more effective than EPA at preventing WD-induced effects on hepatic transcripts linked to fibrosis, including collagen 1A1 (Col1A1), transforming growth factor-β (TGFβ) signaling and proteins involved in remodeling the extracellular matrix, including metalloproteases, tissue inhibitors of metalloproteases, and lysyl oxidase subtypes. Examination of the TGFβ pathway showed that mice fed the WD supplemented with either olive oil or EPA had a significant (≥2.5-fold) increase in hepatic nuclear abundance of phospho-mothers against decapentaplegic homolog (Smad)3 when compared with mice fed the reference diet (RD); Smad3 is a key regulator of Col1A1 expression in stellate cells. In contrast, mice fed the WD supplemented with DHA had no increase in phospho-Smad3 when compared with mice fed the RD. Changes in hepatic phospho-Smad3 nuclear content correlated with proCol1A1 mRNA and protein abundance. Pretreatment of human LX2 stellate cells with DHA, but not other unsaturated fatty acids, blocked TGFβ1-mediated induction of Col1A1. In conclusion, DHA attenuates WD-induced fibrosis by targeting the TGFβ-Smad3-Col1A1 pathway in stellate cells.


Nanophase-separated Ni3Nb as an automobile exhaust catalyst.
Catalytic remediation of automobile exhaust has relied on precious metals (PMs) including platinum (Pt). Herein, we report that an intermetallic phase of Ni and niobium (Nb) (i.e., Ni3Nb) exhibits a significantly higher activity than that of Pt for the remediation of the most toxic gas in exhaust (i.e., nitrogen monoxide (NO)) in the presence of carbon monoxide (CO). When subjected to the exhaust-remediation atmosphere, Ni3Nb spontaneously evolves into a catalytically active nanophase-separated structure consisting of filamentous Ni networks (thickness < 10 nm) that are incorporated in a niobium oxide matrix (i.e., NbO x (x < 5/2)). The exposure of the filamentous Ni promotes NO dissociation, CO oxidation and N2 generation, and the NbO x matrix absorbs excessive nitrogen adatoms to retain the active Ni0 sites at the metal/oxide interface. Furthermore, the NbO x matrix immobilizes the filamentous Ni at elevated temperatures to produce long-term and stable catalytic performance over hundreds of hours.


Intrauterine insemination in natural cycle may give better results in older women.
Controlled ovarian hyper-stimulation (COH) in combination with intrauterine insemination (IUI) has been shown to result in significantly higher pregnancy rates compared to un-stimulated (natural cycle) IUI. This may however not be true in all ages. We performed a retrospective cohort study and analysed data collected prospectively on 1759 IUI cycles in couples with unexplained infertility. The results were analysed to show the outcome of IUI with COH, and IUI in natural cycle (unstimulated), in younger women compared to their older counterparts. In women age 37 and younger, COH resulted in a significantly higher pregnancy rate (13.0% vs 6.5%) and live-birth rate (10.7% vs 5.2%) compared to natural cycle IUI (p = 0.025, p = 0.045 respectively). However for older women age >37 years, natural cycle (unstimulated) IUI, resulted in a significantly higher pregnancy rate (12.0% vs 8.5%) live-birth rate (7.5%vs 3.5%) than IUI with COH ((p = 0.0037). This difference is even more significant when COH was performed with clomiphene citrate (7.5% vs 2.1%) (p = 0.0017). COH was associated with a lower live birth rate in older women, irrespective of the agent used, and it seems to be worse when the anti-oestrogenic drug clomiphene citrate was used for COH. Older women may benefit more from natural cycle (unstimulated) IUI. A randomised controlled trial is required to confirm this observation.


Consequences of early chilling stress in two Triticum species: plastic responses and adaptive significance.
Phenotypic plasticity of two primitive wheat species (Triticum monococcum L. and Triticum dicoccum S.) was studied in response to early chilling stress. Selection pressure differentials, gradients and plasticity costs on plant morphogenesis, growth and reserve carbohydrate consumption were estimated. Regression analysis was applied to investigate differential developmental changes and patterns between treatments. Four-day-old seedlings of T. monococcum and T. dicoccum, differing in plant stature and reserve carbohydrates, were given an early chilling temperature (4 °C for 42 day) and compared with control plants grown at 23 °C. Early chilling stress resulted in a significant increase in leaf mass ratio (LMR) and relative growth rate (RGR), a reduction in flag leaf size, total biomass, specific leaf area (SLA) and reserve carbohydrate storage at flowering, together with advanced onset of flowering. Selection pressure within the early chilling environment favoured early flowering, smaller SLA, higher LMR and lower reserve carbohydrates, suggesting the observed responses were adaptive. Furthermore, a regression of daily cumulative plant biomass derived from a crop growth simulation model (CERES-Wheat) on crop vegetation period revealed a divergent developmental pattern in early-chilled plants. Using selection pressure gradient analysis, we found similar responses among these traits, except for SLA and sucrose, indicating that these two traits have indirect effects on fitness. Thus, the total effects of SLA and reserve sucrose on relative fitness seem to be buffered via the rapid growth rate in chilled plants. While lower SLA may reduce early chilling stress effects at an individual leaf level, a higher LMR and use of reserve carbohydrates indicated that compensatory growth of chilled plants during the recovery period relied on the concerted action of altered resource allocation and reserve carbohydrate consumption. However, a significant cost of plasticity was evident only for flowering time, LMR and fructan levels in the early chilling environment. Our results demonstrate that morphological and intrinsic developmental (ontogenetic) patterns in two Triticum species respond to early chilling stress.


Chlamydia pneumoniae and the lung.
Chlamydia pneumoniae is a frequently occurring respiratory pathogen affecting all age groups. It may cause 5-20% of community-acquired pneumonias in adults and children. The organism has also been implicated as an infectious trigger for asthma. Furthermore, new studies suggest that it may play a role in the pathogenesis of several chronic diseases including atherosclerosis. However, despite the growing significance of C. pneumoniae as a pathogen, progress is hampered by the lack of standardized diagnostic methods including serology and polymerase chain reaction. This makes it practically impossible for the practitioner to make a specific microbiological diagnosis. The lack of standardized methods has also had an adverse effect on treatment trials. The dependence on serology for diagnosis in treatment studies has generated some questionable results. Unless cultures are performed, microbiological efficacy cannot be assessed and it may never be possible to survey for or document the emergence of resistance.


A Procedural Framework for Benchmarking Biofoundry Capabilities.
Benchmarking compares the performance of a product or service with a competitor. In a biofoundry context, capability benchmarking enables more effective use of development resources and furthering business development efforts. Biofoundries considering benchmarking activities are immediately faced with many implementation questions and decisions. While differing circumstances between biofoundries may lead to different answers to those same questions, a common framework for the benchmarking process is desirable. Perhaps the framework described here, and developed for the United States Department of Energy Agile BioFoundry, will be useful to other biofoundries around the world.


HIV-related stigma, social norms, and HIV testing in Soweto and Vulindlela, South Africa: National Institutes of Mental Health Project Accept (HPTN 043).
HIV testing is necessary to curb the increasing epidemic. However, HIV-related stigma and perceptions of low likelihood of societal HIV testing may reduce testing rates. This study aimed to explore this association in South Africa, where HIV rates are extraordinarily high. Data were taken from the Soweto and Vulindlela, South African sites of Project Accept, a multinational HIV prevention trial. Self-reported HIV testing, stigma, and social norms items were used to study the relationship between HIV testing, stigma, and perceptions about societal testing rates. The stigma items were broken into 3 factors: negative attitudes, negative perceptions about people living with HIV, and perceptions of fair treatment for people living with HIV (equity). Results from a univariate logistic regression suggest that history of HIV testing was associated with decreased negative attitudes about people living with HIV/AIDS, increased perceptions that people living with HIV/AIDS experience discrimination, and increased perceptions that people with HIV should be treated equitably. Results from a multivariate logistic regression confirm these effects and suggest that these differences vary according to sex and age. Compared with people who had never tested for HIV, those who had previously tested were more likely to believe that the majority of people have tested for HIV. Data suggest that interventions designed to increase HIV testing in South Africa should address stigma and perceptions of societal testing.


Effect of vaccine dose on the safety and immunogenicity of a candidate TB vaccine, MVA85A, in BCG vaccinated UK adults.
A non-randomised, open-label, Phase I safety and immunogenicity dose-finding study to assess the safety and immunogenicity of the candidate TB vaccine Modified Vaccinia virus Ankara expressing Antigen 85A (MVA85A) from Mycobacterium tuberculosis (MTB) in healthy adult volunteers previously vaccinated with BCG. Healthy BCG-vaccinated volunteers were vaccinated with either 1×10(7) or 1×10(8)PFU of MVA85A. All adverse events were documented and antigen specific T cell responses were measured using an ex vivo IFN-γ ELISPOT assay. Safety and immunogenicity were compared between the 2 dose groups and with a previous trial in which a dose of 5×10(7)PFU MVA85A had been administered. There were no serious adverse events recorded following administration of either 1×10(7) or 1×10(8)PFU of MVA85A. Systemic adverse events were more frequently reported following administration of 1×10(8)PFU of MVA85A when compared to either 5×10(7) or 1×10(7)PFU of MVA85A but were mild or moderate in severity and resolved completely within 7 days of immunisation. Antigen specific T cell responses as measured by the IFN-γ ELISPOT were significantly higher following immunisation in adults receiving 1×10(8)PFU compared to the 5×10(7) and 1×10(7) doses. Additionally, a broader range of Ag85A epitopes are detected following 1×10(8)PFU of MVA85A. A higher dose of 1×10(8)PFU of MVA85A is well-tolerated, increases the frequency of IFN-γ secreting T cells detected following immunisation and broadens the range of Ag85A epitopes detected.


Calpain-10 interacts with plasma saturated fatty acid concentrations to influence insulin resistance in individuals with the metabolic syndrome.
Calpain-10 protein (intracellular Ca(2+)-dependent cysteine protease) may play a role in glucose metabolism, pancreatic β cell function, and regulation of thermogenesis. Several CAPN10 polymorphic sites have been studied for their potential use as risk markers for type 2 diabetes and the metabolic syndrome (MetS). Fatty acids are key metabolic regulators that may interact with genetic factors and influence glucose metabolism. The objective was to examine whether the genetic variability at the CAPN10 gene locus is associated with the degree of insulin resistance and plasma fatty acid concentrations in subjects with MetS. The insulin sensitivity index, glucose effectiveness, insulin resistance [homeostasis model assessment of insulin resistance (HOMA-IR)], insulin secretion (disposition index, acute insulin response, and HOMA of β cell function), plasma fatty acid composition, and 5 CAPN10 single nucleotide polymorphisms (SNPs) were determined in a cross-sectional analysis of 452 subjects with MetS participating in the LIPGENE dietary intervention cohort. The rs2953171 SNP interacted with plasma total saturated fatty acid (SFA) concentrations, which were significantly associated with insulin sensitivity (P < 0.031 for fasting insulin, P < 0.028 for HOMA-IR, and P < 0.012 for glucose effectiveness). The G/G genotype was associated with lower fasting insulin concentrations, lower HOMA-IR, and higher glucose effectiveness in subjects with low SFA concentrations (below the median) than in subjects with the minor A allele (G/A and A/A). In contrast, subjects with the G/G allele with the highest SFA concentrations (above the median) had higher fasting insulin and HOMA-IR values and lower glucose effectiveness than did subjects with the A allele. The rs2953171 polymorphism at the CAPN10 gene locus may influence insulin sensitivity by interacting with the plasma fatty acid composition in subjects with MetS. This trial was registered at clinicaltrials.gov as NCT00429195.


The anti-apoptotic activity of XIAP is retained upon mutation of both the caspase 3- and caspase 9-interacting sites.
The X-linked mammalian inhibitor of apoptosis protein (XIAP) has been shown to bind several partners. These partners include caspase 3, caspase 9, DIABLO/Smac, HtrA2/Omi, TAB1, the bone morphogenetic protein receptor, and a presumptive E2 ubiquitin-conjugating enzyme. In addition, we show here that XIAP can bind to itself. To determine which of these interactions are required for it to inhibit apoptosis, we generated point mutant XIAP proteins and correlated their ability to bind other proteins with their ability to inhibit apoptosis. partial differential RING point mutants of XIAP were as competent as their full-length counterparts in inhibiting apoptosis, although impaired in their ability to oligomerize with full-length XIAP. Triple point mutants, unable to bind caspase 9, caspase 3, and DIABLO/HtrA2/Omi, were completely ineffectual in inhibiting apoptosis. However, point mutants that had lost the ability to inhibit caspase 9 and caspase 3 but retained the ability to inhibit DIABLO were still able to inhibit apoptosis, demonstrating that IAP antagonism is required for apoptosis to proceed following UV irradiation.


The role of amylin in the control of energy homeostasis.
Amylin is an important player in the control of nutrient fluxes. Amylin reduces eating via a meal size effect by promoting meal-ending satiation. This effect seems to depend on a direct action in the area postrema (AP), which is an area rich in amylin receptors. Subsequent to the activation of AP neurons, the neural signal is conveyed to the forebrain via relays involving the nucleus of the solitary tract (NTS) and the lateral parabrachial nucleus (lPBN) to the lateral hypothalamic area (LHA) and other hypothalamic nuclei. While the NTS and lPBN seem to be necessary for amylin's eating inhibitory effect, the role of the LHA has not yet been fully investigated. Amylin may also act as an adiposity signal. Plasma levels of amylin are higher in obese individuals, and chronic infusion of amylin into the brain reduces body weight gain and adiposity; chronic infusion of an amylin receptor antagonist into the brain increases body adiposity. Amylin increases energy expenditure in rats; this effect occurs under various experimental conditions after peripheral and central administration. Together, these animal data, but also clinical data in humans, indicate that amylin is a promising candidate for the treatment of obesity; effects are most pronounced when amylin is combined with leptin. Finally, recent findings indicate that amylin acts as a neurotrophic factor in specific brain stem areas. Whether this effect may be relevant under physiological conditions requires further studies.


Generalized approach for modeling minimally invasive surgery as a stochastic process using a discrete Markov model.
Minimally invasive surgery (MIS) involves a multidimensional series of tasks requiring a synthesis between visual information and the kinematics and dynamics of the surgical tools. Analysis of these sources of information is a key step in defining objective criteria for characterizing surgical performance. The Blue DRAGON is a new system for acquiring the kinematics and the dynamics of two endoscopic tools synchronized with the endoscopic view of the surgical scene. Modeling the process of MIS using a finite state model [Markov model (MM)] reveals the internal structure of the surgical task and is utilized as one of the key steps in objectively assessing surgical performance. The experimental protocol includes tying an intracorporeal knot in a MIS setup performed on an animal model (pig) by 30 surgeons at different levels of training including expert surgeons. An objective learning curve was defined based on measuring quantitative statistical distance (similarity) between MM of experts and MM of residents at different levels of training. The objective learning curve was similar to that of the subjective performance analysis. The MM proved to be a powerful and compact mathematical model for decomposing a complex task such as laparoscopic suturing. Systems like surgical robots or virtual reality simulators in which the kinematics and the dynamics of the surgical tool are inherently measured may benefit from incorporation of the proposed methodology.


Physical activity as a determinant of fasting and 2-h post-challenge glucose: a prospective cohort analysis of the NAVIGATOR trial.
To investigate whether previous physical activity levels are associated with blood glucose levels in individuals with impaired glucose tolerance in the context of an international pharmaceutical trial. Data were analysed from the NAVIGATOR trial, which involved 9306 individuals with impaired glucose tolerance and high cardiovascular risk from 40 different countries, recruited in the period 2002-2004. Fasting glucose, 2-h post-challenge glucose and physical activity (pedometer) were assessed annually. A longitudinal regression analysis was used to determine whether physical activity levels 2 years (t-2 ) and 1 year (t-1 ) previously were associated with levels of glucose, after adjusting for previous glucose levels and other patient characteristics. Those participants with four consecutive annual measures of glucose and two consecutive measures of physical activity were included in the analysis. The analysis included 3964 individuals. Change in physical activity from t-2 to t-1 and activity levels at t-2 were both associated with 2-h glucose levels after adjustment for previous glucose levels and baseline characteristics; however, the associations were weak: a 100% increase in physical activity was associated with a 0.9% reduction in 2-h glucose levels. In addition, previous physical activity only explained an additional 0.05% of the variance in 2-h glucose over the variance explained by the history of 2-h glucose alone (R(2) = 0.3473 vs. 0.3468). There was no association with fasting glucose. In the context of a large international clinical trial, previous physical activity levels did not meaningfully influence glucose levels in those with a high risk of chronic disease, after taking into account participants' previous trajectory of glucose control.


Symptoms of mood disorders in family carers of older people with dementia who experience caregiver burden: a network approach.
informal carers of people with dementia are at greater risk of anxiety and depressive disorders if they find caregiving to be a burden. The aim of this study was to use a network analysis of cross-sectional data to investigate the relationships between anxiety and depressive symptoms in family carers of older people with dementia who experience burden. sixty family carers exhibiting high levels of burden using the Zarit Burden Interview were included in the study. Participants completed the Hospital Anxiety and Depression Scale. The network analysis identified the depression and anxiety symptom network using features including a topological graph, network centrality metrics and community analysis. The network was estimated through the graphical LASSO technique in combination with a walktrap algorithm to obtain the clusters within the network and the connections between the nodes (symptoms). A directed acyclic graph was generated to model symptom interactions. the resulting network architecture shows important bridges between depression and anxiety symptoms. Lack of pleasure and loss of enjoyment were identified as potential gateway symptoms to other anxiety and depression symptoms and represent possible therapeutic targets for psychosocial interventions. Fear and loss of optimism were highly central symptoms, indicating their importance as warning signs of more generalised anxiety and depression. this network analysis of depressive and anxiety symptoms in overburdened family carers provides important insights as to what symptoms may be the most important targets for behavioural interventions.


Transcriptional transactivation by selected short random peptides attached to lexA-GFP fusion proteins.
Transcriptional transactivation is a process with remarkable tolerance for sequence diversity and structural geometry. In studies of the features that constitute transactivating functions, acidity has remained one of the most common characteristics observed among native activation domains and activator peptides. We performed a deliberate search of random peptide libraries for peptides capable of conferring transcriptional transactivation on the lexA DNA binding domain. Two libraries, one composed of C-terminal fusions, the other of peptide insertions within the green fluorescent protein structure, were used. We show that (i) peptide sequences other than C-terminal fusions can confer transactivation; (ii) though acidic activator peptides are more common, charge neutral and basic peptides can function as activators; and (iii) peptides as short as 11 amino acids behave in a modular fashion. These results support the recruitment model of transcriptional activation and, combined with other studies, suggest the possibility of using activator peptides in a variety of applications, including drug development work.


Morphological recovery in the reattached retina.
After experimental retinal detachment in the cat, a number of morphological changes take place in retinal and RPE cells. Following reattachment, the ultrastructural relationship between the photoreceptors and the RPE is re-established, but it does not return to the predetachment state even after short detachment episodes coupled with prolonged recovery periods. All of the reattached retinae show some degree of abnormality, ranging from subtle changes in photoreceptor ultrastructure to dramatic degenerative effects in the outer retina. Abrupt transitions in morphology from one reattached area to an adjacent area are not unusual. Photoreceptor recovery varies widely between animals, and between adjacent regions within the same retina. Ensheathment of outer segments by RPE apical processes is abnormal. In some reattached areas rod outer segment dimensions and disc structure are near normal as is the displacement rate of rod outer segment discs. In others, especially in areas of RPE or Müller cell proliferation and hypertrophy, the outer segments are shortened or absent completely, and there is a reduction of cell bodies in the outer nuclear layer. In some retinae, recovery in cones is inferior to that in rods. At short detachment durations (less than 1 wk) morphological recovery in the reattached retina is optimal while at long intervals (greater than 1 month) recovery is poor. The changes at the photoreceptor-RPE interface identified in the reattached cat retina probably have adverse effects on visual recovery when they occur within the human macula.


Global adaptation in networks of selfish components: emergent associative memory at the system scale.
In some circumstances complex adaptive systems composed of numerous self-interested agents can self-organize into structures that enhance global adaptation, efficiency, or function. However, the general conditions for such an outcome are poorly understood and present a fundamental open question for domains as varied as ecology, sociology, economics, organismic biology, and technological infrastructure design. In contrast, sufficient conditions for artificial neural networks to form structures that perform collective computational processes such as associative memory/recall, classification, generalization, and optimization are well understood. Such global functions within a single agent or organism are not wholly surprising, since the mechanisms (e.g., Hebbian learning) that create these neural organizations may be selected for this purpose; but agents in a multi-agent system have no obvious reason to adhere to such a structuring protocol or produce such global behaviors when acting from individual self-interest. However, Hebbian learning is actually a very simple and fully distributed habituation or positive feedback principle. Here we show that when self-interested agents can modify how they are affected by other agents (e.g., when they can influence which other agents they interact with), then, in adapting these inter-agent relationships to maximize their own utility, they will necessarily alter them in a manner homologous with Hebbian learning. Multi-agent systems with adaptable relationships will thereby exhibit the same system-level behaviors as neural networks under Hebbian learning. For example, improved global efficiency in multi-agent systems can be explained by the inherent ability of associative memory to generalize by idealizing stored patterns and/or creating new combinations of subpatterns. Thus distributed multi-agent systems can spontaneously exhibit adaptive global behaviors in the same sense, and by the same mechanism, as with the organizational principles familiar in connectionist models of organismic learning.


Coupled RNA polymerase II transcription and 3' end formation with yeast whole-cell extracts.
RNA polymerase II (RNAP II) transcription and pre-mRNA 3' end formation are linked through physical and functional interactions. We describe here a highly efficient yeast in vitro system that reproduces both transcription and 3' end formation in a single reaction. The system is based on simple whole-cell extracts that were supplemented with a hybrid Gal4-VP16 transcriptional activator and supercoiled plasmid DNA templates encoding G-less cassette reporters. We found that the coupling of transcription and processing in vitro enhanced pre-mRNA 3' end formation and reproduced requirements for poly(A) signals and polyadenylation factors. Unexpectedly, however, we show that in vitro transcripts lacked m⁷G-caps. Reconstitution experiments with CF IA factor assembled entirely from heterologous components suggested that the CTD interaction domain of the Pcf11 subunit was required for proper RNAP II termination but not 3' end formation. Moreover, we observed reduced termination activity associated with extracts prepared from cells carrying a mutation in the 5'-3' exonuclease Rat1 or following chemical inhibition of exonuclease activity. Thus, in vitro transcription coupled to pre-mRNA processing recapitulates hallmarks of poly(A)-dependent RNAP II termination. The in vitro transcription/processing system presented here should provide a useful tool to further define the role of factors involved in coupling.


Cloning, expression, purification, crystallization and preliminary X-ray diffraction studies of a 12R-LOX-chaperone complex.
Lipoxygenases are a family of nonheme iron-containing dioxygenases. An Escherichia coli expression system producing the bacterial chaperones GroES and GroEL was engineered and successfully used to produce large quantities of recombinant human 12R-LOX (LOXR; MW 80.34 kDa; 701 amino-acid residues). The co-overproduction of the two chaperones with 12R-LOX resulted in increased solubility of 12R-LOX and allowed the purification of milligram amounts of active enzyme for structural studies by X-ray diffraction. The lipoxygenase protein was purified on an affinity column and a gel-filtration column with chaperone protein (MW 57.16 kDa). The LOXR-chaperone complex was crystallized with ligand by the hanging-drop vapor-diffusion method using 1.5 M ammonium hydrogen phosphate as precipitant. The crystals belonged to the monoclinic system, space group P2(1), with unit-cell parameters a = 138.97, b = 266.11, c = 152.26 Å, β = 101.07°. Based on the calculated Matthews coefficient (3.1 Å(3) Da(-1)), it is estimated that one molecule of LOXR complexed with two molecules of chaperone is present in the asymmetric unit of the crystal lattice. X-ray diffraction data were collected to 4 Å resolution using synchrotron radiation.


Insulin-like growth factor 1 synergizes with bone morphogenetic protein 7-mediated anabolism in bovine intervertebral disc cells.
We undertook this study to assess the therapeutic benefits of intervertebral disc matrix repair and regeneration by evaluating the potential synergistic effect of insulin-like growth factor 1 (IGF-1) and bone morphogenetic protein 7 (BMP-7) on bovine spine discs and by elucidating the relevant molecular/cellular mechanisms. Bovine nucleus pulposus (NP) cells were treated with BMP-7 and IGF-1. The subsequent anabolic effects driven by NP cells were assessed for proteoglycan (PG) synthesis by (35) S-sulfate incorporation and for PG accumulation by dimethylmethylene blue assays. Matrix formation was visualized by particle exclusion assay. Key matrix components and transcription factors were analyzed by real-time reverse transcription-polymerase chain reaction to determine the signaling pathways by which IGF-1 suppresses noggin, a potent inhibitor of BMP-7. Western blotting and nuclear translocation experiments were performed to assess the activation of Smad proteins. Stimulation of bovine NP cells by both IGF-1 and BMP-7 greatly potentiated anabolism through complementary and synergistic mechanisms on matrix formation compared with treatment with either growth factor alone. The exogenously added decoy ligand, noggin, attenuated the anabolic effects of BMP-7, and noggin was substantially increased by BMP-7, suggesting a negative feedback regulatory mechanism. In contrast, IGF-1 significantly suppressed noggin expression via the phosphatidylinositol 3-kinase/Akt pathway and thus potentiated BMP-7 signaling in bovine NP cells. Upon combination treatment, IGF-1 activated Smad2, while BMP-7 activated Smad1/5/8 and Smad3, thus inducing all Smad signaling pathways and mimicking the effects of the combination of transforming growth factor β and BMP-7 Combination growth factor therapy using BMP-7 and IGF-1 may have considerable promise in the treatment of spine disc degeneration.


In vivo near-realtime volumetric optical-resolution photoacoustic microscopy using a high-repetition-rate nanosecond fiber-laser.
Optical-resolution photoacoustic microscopy (OR-PAM) is capable of achieving optical-absorption-contrast images with micron-scale spatial resolution. Previous OR-PAM systems have been frame-rate limited by mechanical scanning speeds and laser pulse repetition rate (PRR). We demonstrate OR-PAM imaging using a diode-pumped nanosecond-pulsed Ytterbium-doped 532-nm fiber laser with PRR up to 600 kHz. Combined with fast-scanning mirrors, our proposed system provides C-scan and 3D images with acquisition frame rate of 4 frames per second (fps) or higher, two orders of magnitude faster than previously published systems. High-contrast images of capillary-scale microvasculature in a live Swiss Webster mouse ear with ~6-µm optical lateral spatial resolution are demonstrated.


[Mobile miniature X-ray evaluation and pneumoconiosis: the role of the Clinica del Lavoro in Milan (1941-1948)].
Since the end of the 19th century, X-rays have been used to detect lung diseases. In Italy, 207,096 miniature chest radiographs were taken from 1941 to 1948. Traditional radiographs gave better results, but miniature chest radiographs were useful for screening. Indeed, the development of mobile miniature chest radiography units resulted in an improvement in mass X-rays screening for the detection of penumoconiosis. These mobile miniature units were mounted on a bus chassis, a solution that allowed to easily reach workers. The authors analyze some models of X-ray wagon units used by the "Clinica del Lavoro" in Milan in the 1950s. From the point of view of medical museology, the preservation of these devices requires appropriate spaces.


Inferring Connectivity Range in Submerged Aquatic Populations (Ruppia L.) Along European Coastal Lagoons From Genetic Imprint and Simulated Dispersal Trajectories.
Coastal salt- and brackish water lagoons are unique shallow habitats characterized by beds of submerged seagrasses and salt-tolerant Ruppia species. Established long-term and large-scale patterns of connectivity in lagoon systems can be strongly determined by patterns of nearshore and coastal currents next to local bird-mediated seed dispersal. Despite the importance of dispersal in landscape ecology, characterizing patterns of connectivity remains challenging in aquatic systems. Here, we aimed at inferring connectivity distances of Ruppia cirrhosa along European coastal lagoons using a population genetic imprint and modeled dispersal trajectories using an eddy-resolving numerical ocean model that includes tidal forcing. We investigated 1,303 individuals of 46 populations alongside subbasins of the Mediterranean (Balearic, Tyrrhenian, Ionian) and the Atlantic to Baltic Sea coastline over maximum distances of 563-2,684 km. Ten microsatellite loci under an autotetraploid condition revealed a mixed sexual and vegetative reproduction mode. A pairwise FST permutation test of populations revealed high levels of historical connectivity only for distance classes up to 104-280 km. Since full range analysis was not fully explanatory, we assessed connectivity in more detail at coastline and subbasin level using four approaches. Firstly, a regression over restricted geographical distances (300 km) was done though remained comparable to full range analysis. Secondly, piecewise linear regression analyses yielded much better explained variance but the obtained breakpoints were shifted toward greater geographical distances due to a flat slope of regression lines that most likely reflect genetic drift. Thirdly, classification and regression tree analyses revealed threshold values of 47-179 km. Finally, simulated ocean surface dispersal trajectories for propagules with floating periods of 1-4 weeks, were congruent with inferred distances, a spatial Bayesian admixed gene pool clustering and a barrier detection method. A kinship based spatial autocorrelation showed a contemporary within-lagoon connectivity up to 20 km. Our findings indicate that strong differentiation or admixtures shaped historical connectivity and that a pre- and post LGM genetic imprint of R. cirrhosa along the European coasts was maintained from their occurrence in primary habitats. Additionally, this study demonstrates the importance of unraveling thresholds of genetic breaks in combination with ocean dispersal modeling to infer patterns of connectivity.


Denaturalizing scarcity: a strategy of enquiry for public- health ethics.
Most scarcities that underpin health disparities within and among countries are not natural; rather, they result from policy choices and the operation of social institutions. Using examples from the United States of America: the Chicago heat wave and hurricane Katrina, this paper develops "denaturalizing scarcity" as a strategy for enquiry to inform public-health ethics in an interconnected world. It first describes some of the resource scarcities that are of greatest concern from a public-health perspective, and then outlines two (not mutually exclusive) lines of ethical reasoning that demonstrate their importance. One of these involves the multiple relationships that link rich and poor across national borders in today's interconnected world. The paper then briefly describes ways in which globalization and the associated institutions are linked to health-threatening scarcities. The paper concludes that denaturalizing scarcity represents a valuable alternative to mainstream health ethics, directing our attention instead to why some settings are "resource poor" and others are not.


Gene expression profiling of peripheral blood from patients with untreated new-onset systemic juvenile idiopathic arthritis reveals molecular heterogeneity that may predict macrophage activation syndrome.
Systemic juvenile idiopathic arthritis (JIA) is frequently associated with the development of macrophage activation syndrome. This study was undertaken to better understand the relationship between systemic JIA and macrophage activation syndrome. Gene expression profiles were examined in 17 patients with untreated new-onset systemic JIA, 5 of whom showed evidence of subclinical macrophage activation syndrome (of whom 2 eventually developed overt macrophage activation syndrome). Peripheral blood mononuclear cells (PBMCs) were separated on Ficoll gradients, and purified RNA was analyzed using Affymetrix GeneChip expression arrays. A fraction of the PBMCs were used for flow cytometry to define the cellular composition of the samples. Two hundred twenty-five differentially expressed genes (P < 0.05) that distinguished patients with systemic JIA from healthy controls (n = 30) were identified. Clustering analysis indicated that expression patterns correlated with serum ferritin levels. Three main clusters distinguished systemic JIA patients with highly elevated ferritin levels (including those with subclinical macrophage activation syndrome) from those with normal or only moderately elevated ferritin levels. The first cluster comprised genes involved in the synthesis of hemoglobins and structural proteins of erythrocytes. This transcriptional profile was consistent with immature nucleated red blood cells, likely reflective of high red blood cell turnover. Also included were transcripts indicating immature granulocytes. The second cluster was enriched for genes involved in cell cycle regulation. The third cluster was enriched for genes involved in innate immune responses, including those involved in the negative regulation of Toll-like receptor/interleukin-1 receptor-triggered inflammatory cascades and markers of the alternative pathway of macrophage differentiation. Additional differentially expressed genes of interest were those involved in the cytolytic pathway, including SH2D1A and Rab27a. These data indicate that gene expression profiling can be a useful tool for identifying early macrophage activation syndrome in patients with systemic JIA.


Using realist review to inform intervention development: methodological illustration and conceptual platform for collaborative care in offender mental health.
This paper reports how we used a realist review, as part of a wider project to improve collaborative mental health care for prisoners with common mental health problems, to develop a conceptual platform. The importance of offenders gaining support for their mental health, and the need for practitioners across the health service, the criminal justice system, and the third sector to work together to achieve this is recognised internationally. However, the literature does not provide coherent analyses of how these ambitions can be achieved. This paper demonstrates how a realist review can be applied to inform complex intervention development that spans different locations, organisations, professions, and care sectors. We applied and developed a realist review for the purposes of intervention development, using a three-stage process. (1) An iterative database search strategy (extending beyond criminal justice and offender health) and groups of academics, practitioners, and people with lived experience were used to identify explanatory accounts (n = 347). (2) From these accounts, we developed consolidated explanatory accounts (n = 75). (3) The identified interactions between practitioners and offenders (within their organisational, social, and cultural contexts) were specified in a conceptual platform. We also specify, step by step, how these explanatory accounts were documented, consolidated, and built into a conceptual platform. This addresses an important methodological gap for social scientists and intervention developers about how to develop and articulate programme and implementation theory underpinning complex interventions. An integrated person-centred system is proposed to improve collaborative mental health care for offenders with common mental health problems (near to and after release) by achieving consistency between the goals of different sectors and practitioners, enabling practitioners to apply scientific and experiential knowledge in working judiciously and reflectively, and building systems and aligning resources that are centred on offenders' health and social care needs. As part of a broader programme of work, a realist review can make an important contribution to the specification of theoretically informed interventions that have the potential to improve health outcomes. Our conceptual platform has potential application in related systems of health and social care where integrated, and person-centred care is a goal.


Multi-level considerations for optimal implementation of long-acting injectable antiretroviral therapy to treat people living with HIV: perspectives of health care providers participating in phase 3 trials.
Long-acting injectable antiretroviral therapy (LA ART) has been shown to be non-inferior to daily oral ART, with high patient satisfaction and preference to oral standard of care in research to date, and has recently been approved for use in the United States and Europe. This study examined the perspectives of health care providers participating in LA ART clinical trials on potential barriers and solutions to LA ART roll-out into real world settings. This analysis draws on two data sources: (1) open-ended questions embedded in a structured online survey of 329 health care providers participating in the ATLAS-2 M trial across 13 countries; and (2) in-depth interviews with 14 providers participating in FLAIR/ ATLAS/ATLAS-2 M trials in the United States and Spain. Both assessments explored provider views and clinic dynamics related to the introduction of LA ART and were analyzed using thematic content analysis. The Consolidated Framework for Implementation Research (CFIR) was drawn on as the conceptual framework underpinning development of a model depicting study findings. Barriers and proposed solutions to LA ART implementation were identified at the individual, clinic and health system levels. Provider perceptions of patient level barriers included challenges with adhering to frequent injection appointments and injection tolerability. Proposed solutions included patient education, having designated staff for clinic visit retention, and clinic flexibility with appointment scheduling. The main provider concern was identifying appropriate candidates for LA ART; proposed solutions focused on patient provider communication and decision making. Clinic level barriers included the need for additional skilled individuals to administer injections, shifts in workflow as demand increases and the logistics of cold-chain storage. Proposed solutions included staff hiring and training, strategic planning around workflow and logistics, and the possibility of offering injections in other settings, including the home. Health system level barriers included cost and approvals from national regulatory bodies. Potential solutions included governments subsidizing treatment, ensuring cost is competitive with oral ART, and offering co-pay assistance. Results suggest the importance of multi-level support systems to optimize patient-provider communication and treatment decision-making; clinic staffing, workflow, logistics protocols and infrastructure; and cost-related factors within a given health system.


Building blocks, cohort genetic algorithms, and hyperplane-defined functions.
Building blocks are a ubiquitous feature at all levels of human understanding, froin perception through science and innovation. Genetic algorithms are designed to exploit this prevalence. A new, more robust class of genetic algorithms, cohort genetic algorithms (cGA's), provides substantial advantages in exploring search spaces for building blocks while exploiting building blocks already found. To test these capabilities, a new, general class of test functions, the hyperplane-defined functions (hdf's), has been designed. Hdf's offer the means of tracing the origin of each advance in performance; at the same time hdf's are resistant to reverse engineering, so that algorithms cannot be designed to take advantage of the characteristics of particular examples.


Daily text messaging for weight control among racial and ethnic minority women: randomized controlled pilot study.
Daily self-monitoring of diet and physical activity behaviors is a strong predictor of weight loss success. Text messaging holds promise as a viable self-monitoring modality, particularly among racial/ethnic minority populations. This pilot study evaluated the feasibility of a text messaging intervention for weight loss among predominantly black women. Fifty obese women were randomized to either a 6-month intervention using a fully automated system that included daily text messages for self-monitoring tailored behavioral goals (eg, 10,000 steps per day, no sugary drinks) along with brief feedback and tips (n=26) or to an education control arm (n=24). Weight was objectively measured at baseline and at 6 months. Adherence was defined as the proportion of text messages received in response to self-monitoring prompts. The average daily text messaging adherence rate was 49% (SD 27.9) with 85% (22/26) texting self-monitored behavioral goals 2 or more days per week. Approximately 70% (16/23) strongly agreed that daily texting was easy and helpful and 76% (16/21) felt the frequency of texting was appropriate. At 6 months, the intervention arm lost a mean of 1.27 kg (SD 6.51), and the control arm gained a mean of 1.14 kg (SD 2.53; mean difference -2.41 kg, 95% CI -5.22 to 0.39; P=.09). There was a trend toward greater text messaging adherence being associated with greater percent weight loss (r=-.36; P=.08), but this did not reach statistical significance. There was no significant association between goal attainment and text messaging adherence and no significant predictors of adherence. Given the increasing penetration of mobile devices, text messaging may be a useful self-monitoring tool for weight control, particularly among populations most in need of intervention. Clinicaltrials.gov: NCT00939081; http://clinicaltrials.gov/show/NCT00939081 (Archived by WebCite at http://www.webcitation.org/6KiIIcnk1).


SARS-CoV-2 Cumulative Incidence and Period Seroprevalence: Results From a Statewide Population-Based Serosurvey in California.
California has reported the largest number of coronavirus disease 2019 (COVID-19) cases of any US state, with more than 3.5 million confirmed as of March 2021. However, the full breadth of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) transmission in California is unknown as reported cases only represent a fraction of all infections. We conducted a population-based serosurvey, utilizing mailed, home-based SARS-CoV-2 antibody testing along with a demographic and behavioral survey. We weighted data from a random sample to represent the adult California population and estimated period seroprevalence overall and by participant characteristics. Seroprevalence estimates were adjusted for waning antibodies to produce statewide estimates of cumulative incidence, the infection fatality ratio (IFR), and the reported fraction. California's SARS-CoV-2 weighted seroprevalence during August-December 2020 was 4.6% (95% CI, 2.8%-7.4%). Estimated cumulative incidence as of November 2, 2020, was 8.7% (95% CrI, 6.4%-11.5%), indicating that 2 660 441 adults (95% CrI, 1 959 218-3 532 380) had been infected. The estimated IFR was 0.8% (95% CrI, 0.6%-1.0%), and the estimated percentage of infections reported to the California Department of Public Health was 31%. Disparately high risk for infection was observed among persons of Hispanic/Latinx ethnicity and people with no health insurance and who reported working outside the home. We present the first statewide SARS-CoV-2 cumulative incidence estimate among adults in California. As of November 2020, ~1 in 3 SARS-CoV-2 infections in California adults had been identified by public health surveillance. When accounting for unreported SARS-CoV-2 infections, disparities by race/ethnicity seen in case-based surveillance persist.


The social determinants of health of the child-adolescent immigration and its influence on the nutritional status: systematic review.
to review the social determinants of health more characteristic of the child and adolescents of immigrants, by reviewing the literature and assess its effect on nutritional status. a systematic review was performed using Medical Subject Headings (MeSH) in PubMed (Medline) and The Cochrane Library, in order to identify undetected studies; articles bibliographic lists were examined. The final election was done according to inclusion and exclusion criteria. No restrictions on sex and ethnicity of the participants. STROBE checkpoints were used for an information and methodological quality control. As Social Determinants of Health (SDH); social, demographic and economic conditions were considered for the study of their effect on the nutritional status. from 31 identified articles 18 are included in this study, 17 (94.4%) had a good or excellent quality. Hispanic and African were the most studied ethnicities; birth place and parent's residence period were used for generational classification. Alimentary culture and linguistic isolation of the first generation have a protective effect preventing from overweight and obesity risk while it decrease in second and third generation due to the experienced acculturation process equalizing their weight gain to natives; which prevalence is higher among Hispanics. No relation was found for nutritional status differences between sexes neither alimentary aids protective effect hypothesis was confirmed. the SDH with greater influence on child-adolescent immigrants were the socio-demographic conditions; among them: residence period distinguish the three identified generations while linguistic barrier and ethnic background are truly influential on the biological response to the experimented change caused by the acculturation process, establishing differences in the nutritional status.


Cognitive assessment across the continuum of care: the importance of occupational performance-based assessment for individuals post-stroke and traumatic brain injury.
When working with individuals following stroke or traumatic brain injury, an important role of the occupational therapist is to assess the impact of cognitive impairment on their ability to engage in occupations and resume important life roles. The aim of this study was to survey therapists' reasons for selection of and challenges with using various cognitive assessment approaches, across the continuum of care, when working with individuals following stroke and traumatic brain injury. A cross-sectional survey, completed via post or online, with responses from 209 Australian occupational therapists was conducted. Participants included clinicians working in acute, inpatient rehabilitation and community settings. Occupational performance-based assessments were ranked as the most important assessment method, with 69% of participants reporting using these assessments for more than 75% of their clients with cognitive impairment. Participants identified the lack of quantitative data provided by these assessments as a frequent challenge. The identification of cognitive deficits was the highest ranked reason for using cognitive screens and batteries. Challenges identified with using cognitive screens and batteries included difficulty linking assessment results to occupational performance, and difficulty using results to generate intervention strategies. The majority of participants reported using a combined approach to assessment, and used screens and batteries to support findings of occupational performance-based assessments. Targeted efforts to further incorporate standardised occupational performance-based methods into clinical practice, research, and ongoing professional development is required to enhance occupational therapy services when working with individuals with cognitive impairment.


Evaluation of a high school peer group intervention for at-risk youth.
The purpose of this paper is to examine the effectiveness of Reconnecting Youth, a prevention program for at-risk high school youth. Data are from a large, independently evaluated effectiveness trial in two diverse urban school districts. A total of 1,218 students participated; 50% were male; average age was 15. We tested whether positive efficacy trial effects could be replicated, and whether any negative behavioral effects occur when clustering high-risk youth. Although mixed program effects were observed at immediate post-intervention, only negative effects were found at 6-month follow-up. These effects included less optimal scores on measures of GPA, Anger, School Connectedness, Conventional Peer Bonding, and Peer High-Risk Behaviors. Overall, we found little support for the use of this social-influence--model intervention aimed at increasing school connectedness for high-risk youth. Further, this study provides evidence that clustering high-risk youth in preventive interventions has the potential for iatrogenic effects.


Radial pressure waves mediate apoptosis and functional angiogenesis during wound repair in ApoE deficient mice.
This study aims to quantify by intravital microscopy and histological wound scoring the effect of radial pressure wave treatment (RPWT) on murine incisional wound healing. The dorsal skinfold chamber in mice was used for intravital microscopy, whereby an incisional wound was created within the chamber. RPWT to the wound was carried out using a ballistic pressure wave source (EMS Swiss DolorClast). Animals received a dose of 500 pulses at an energy flux rate of 0.1mJ/mm(2) and a frequency of 3Hz at day 1, 3, 5, 7, 9, and 11 post wounding. RPW treated and untreated ApoE depleted mice (ApoE(-/-)) were compared to normal healing wild type animals (WT). The microcirculation of the wound was analyzed quantitatively in vivo using epi-illumination intravital fluorescence microscopy. Tissue samples were examined ex vivo for wound scoring and immunohistochemistry. Upon RPWT total wound score in ApoE(-/-) mice was increased by 13% (not significant) on day 3, by 37% on day 7 (P<0.05), and by 39% on day 13 (P<0.05) when compared to untreated ApoE(-/-) mice. Improved wound healing was associated with an increase of functional angiogenetic density by 23% (not significant) on day 5, by 36% on day 7 (P<0.05), and by 41% on day 9 (P<0.05). Following RPWT, on day three we observed enhanced expression of capase-3 (2-fold), proliferating cell nuclear antibody (PCNA, 1,6-fold), and endothelial nitric oxide synthase (eNOS, 2.6-fold), all P<0.05. In conclusion repetitive RPWT accelerated wound healing in ApoE(-/-) mice by increasing functional neovascular density. In addition our findings strongly suggest that RPW may facilitate the linear progression of wound healing phases by fostering apoptosis.


Long noncoding RNA as modular scaffold of histone modification complexes.
Long intergenic noncoding RNAs (lincRNAs) regulate chromatin states and epigenetic inheritance. Here, we show that the lincRNA HOTAIR serves as a scaffold for at least two distinct histone modification complexes. A 5' domain of HOTAIR binds polycomb repressive complex 2 (PRC2), whereas a 3' domain of HOTAIR binds the LSD1/CoREST/REST complex. The ability to tether two distinct complexes enables RNA-mediated assembly of PRC2 and LSD1 and coordinates targeting of PRC2 and LSD1 to chromatin for coupled histone H3 lysine 27 methylation and lysine 4 demethylation. Our results suggest that lincRNAs may serve as scaffolds by providing binding surfaces to assemble select histone modification enzymes, thereby specifying the pattern of histone modifications on target genes.


Lost in translation: reproductive health care experiences of Somali Bantu women in Hartford, Connecticut.
Reproductive health problems are the leading cause of women's morbidity and mortality worldwide. In the United States, officially sponsored refugee women continue to face challenges in accessing reproductive health programs despite having access to health insurance. The objective of this study was to explore the reproductive health experiences of 1 such population--Somali Bantu women in Connecticut--to identify potential barriers to care experienced by marginalized populations. The study was qualitative, consisting of key informant interviews, a focus group session, and a semistructured survey. Although all the women in the study reported having access to reproductive health care services, they also reported having unmet health needs resulting from barriers to care that included ethnic distinction/language barriers, passive acceptance of incorrect care, cultural discordance in family planning services, patient-provider sex discordance, and desire but limited scope for ownership in health care outcomes. The root cause of the various types of patient-provider discordance was the lack of recognition that the Somali Bantu are distinct in culture, language, and solidarity from ethnic Somalis, resulting in Language Line translation services being conducted in a Somali language that the Somali Bantu women did not understand. The results of the study primarily highlight the larger issue of information asymmetry within the health care system that, if left unaddressed, will persist as new vulnerable populations of refugees arrive in the United States.


Is the time right for quantitative public health guidelines on sitting? A narrative review of sedentary behaviour research paradigms and findings.
Sedentary behaviour (SB) has been proposed as an 'independent' risk factor for chronic disease risk, attracting much research and media attention. Many countries have included generic, non-quantitative reductions in SB in their public health guidelines and calls for quantitative SB targets are increasing. The aim of this narrative review is to critically evaluate key evidence areas relating to the development of guidance on sitting for adults. We carried out a non-systematic narrative evidence synthesis across seven key areas: (1) definition of SB, (2) independence of sitting from physical activity, (3) use of television viewing as a proxy of sitting, (4) interpretation of SB evidence, (5) evidence on 'sedentary breaks', (6) evidence on objectively measured sedentary SB and mortality and (7) dose response of sitting and mortality/cardiovascular disease. Despite research progress, we still know little about the independent detrimental health effects of sitting, and the possibility that sitting is mostly the inverse of physical activity remains. Unresolved issues include an unclear definition, inconsistencies between mechanistic and epidemiological studies, over-reliance on surrogate outcomes, a very weak epidemiological evidence base to support the inclusion of 'sedentary breaks' in guidelines, reliance on self-reported sitting measures, and misinterpretation of data whereby methodologically inconsistent associations are claimed to be strong evidence. In conclusion, public health guidance requires a consistent evidence base but this is lacking for SB. The development of quantitative SB guidance, using an underdeveloped evidence base, is premature; any further recommendations for sedentary behaviour require development of the evidence base and refinement of the research paradigms used in the field.


Recognition of an endothelial determinant for CD 18-dependent human neutrophil adherence and transendothelial migration.
Human neutrophil (PMN) attachment to human umbilical vein endothelial cells (HUVEC) was evaluated in vitro using two MAbs, R6-5-D6 and RR1/1, that recognize intercellular adhesion molecule-1 (ICAM-1), and one MAb, TS1/18, that recognizes CD18. Pretreatment of the HUVEC with anti-ICAM-1 MAbs produced greater than 50% inhibition of attachment to HUVEC, and IL-1 (0.5 U/ml)- or lipopolysaccharide (LPS) (10 ng/ml)-stimulated HUVEC, and greater than 99% inhibition of f-Met-Leu-Phe (0.5 nM) enhanced adherence. Anti-ICAM-1 MAbs also inhibited by greater than 85% the transendothelial migration induced by 4-h IL-1 (0.5 U/ml) and LPS (10 ng/ml) activation of the HUVEC. That these effects involved a CD18-dependent mechanism is supported by the following results: pretreatment of PMN with TS1/18 produced the same degree of inhibition of attachment and migration as seen with R6-5-D6. In addition, the use of both MAbs together did not further increase the inhibition of cell attachment to stimulated HUVEC. The attachment of PMN from patients with CD18 deficiency to stimulated HUVEC was not reduced by R6-5-D6, and both R6-5-D6 and TS1/18 revealed the same time course for appearance and disappearance of an adherence component on stimulated HUVEC not blocked by either MAb. These results demonstrate that attachment and transendothelial migration of PMN in vitro depend substantially on both CD18 on the PMN and ICAM-1 on the endothelial cell.


Induction of cell cycle-dependent cytotoxicity and apoptosis by new heterodinucleoside phosphate dimers of 5-fluorodeoxyuridine in PC-3 human prostate cancer cells.
Fluorodeoxyuridine (5-FdUrd) is an antineoplastic agent with clinical activity against different types of solid tumours. To enhance the effectiveness of this drug, we have synthesised new heterodinucleoside phosphate dimers of 5-FdUrd. These dimers were compared to 5-FdUrd for their cytotoxic effect and the cell cycle dependence of cytotoxicity, as well as for their capacity to induce apoptosis and inhibit thymidylate synthetase (TS) in androgen-independent human PC-3 prostate tumour cells. Incubation of the cells with the dimers N(4)-palmitoyl-2'-deoxycytidylyl-(3'-->5')-5-fluoro-2'-deoxyuri din e (dCpam-5-FdUrd) and 2'-deoxy-5-flourouridylyl-(3'-->5')-2'-deoxy-5-fluoro-N(4)-octa decylc ytidine (5-FdUrd-5-FdC18) resulted in a marked cytotoxicity with IC(50) values of 4 microM, similar to 5-FdUrd. In contrast to 5-FdUrd, 100% toxicity was achieved with concentrations of 100-200 microM 5-FdUrd-5-FdC18. Flow cytometric analysis revealed an increase in the cell population in S-phase after treatment with 5-FdUrd, 5-FdUrd-5-FdC18, and dCpam-5-FdUrd from 36 to 63%, 50%, and 77%, respectively. dCpam-5-FdUrd was more potent than 5-FdUrd in arresting the cell cycle. Significant S-phase arrest was indicated by a decreased proportion of cells in G1- and G2/M-phases. Cell cycle arrest and inhibition of cell proliferation were followed by apoptosis, as shown by a 6- to 8-fold increased binding of Apo2.7 antibody, a 9- to 11-fold increase in caspase-3 activity, DNA fragmentation, and by cell morphology showing the appearance of apoptotic bodies. Importantly, 5-FdUrd-5-FdC18 increased the number of apoptotic cells to 160% compared to 5-FdUrd under the same conditions. As with 5-FdUrd, the two dimers also inhibited TS in a time- and concentration-dependent manner, although requiring 100-fold higher concentrations. In conclusion, dCpam-5-FdUrd and 5-FdUrd-5-FdC18 exert stronger cytotoxicity and induce more S-phase arrest and apoptosis than does 5-FdUrd in PC-3 cells, suggesting their potential role in the treatment of human prostate cancer.


Menage a quoi? Optimal number of peer reviewers.
Peer review represents the primary mechanism used by funding agencies to allocate financial support and by journals to select manuscripts for publication, yet recent Cochrane reviews determined literature on peer review best practice is sparse. Key to improving the process are reduction of inherent vulnerability to high degree of randomness and, from an economic perspective, limiting both the substantial indirect costs related to reviewer time invested and direct administrative costs to funding agencies, publishers and research institutions. Use of additional reviewers per application may increase reliability and decision consistency, but adds to overall cost and burden. The optimal number of reviewers per application, while not known, is thought to vary with accuracy of judges or evaluation methods. Here I use bootstrapping of replicated peer review data from a Post-doctoral Fellowships competition to show that five reviewers per application represents a practical optimum which avoids large random effects evident when fewer reviewers are used, a point where additional reviewers at increasing cost provides only diminishing incremental gains in chance-corrected consistency of decision outcomes. Random effects were most evident in the relative mid-range of competitiveness. Results support aggressive high- and low-end stratification or triaging of applications for subsequent stages of review, with the proportion and set of mid-range submissions to be retained for further consideration being dependent on overall success rate.