Structure of mammalian endolysosomal TRPML1 channel in nanodiscs.
Transient receptor potential mucolipin 1 (TRPML1) is a cation channel located within endosomal and lysosomal membranes. Ubiquitously expressed in mammalian cells, its loss-of-function mutations are the direct cause of type IV mucolipidosis, an autosomal recessive lysosomal storage disease. Here we present the single-particle electron cryo-microscopy structure of the mouse TRPML1 channel embedded in nanodiscs. Combined with mutagenesis analysis, the TRPML1 structure reveals that phosphatidylinositol-3,5-bisphosphate (PtdIns(3,5)P2) binds to the N terminus of the channel-distal from the pore-and the helix-turn-helix extension between segments S2 and S3 probably couples ligand binding to pore opening. The tightly packed selectivity filter contains multiple ion-binding sites, and the conserved acidic residues form the luminal Ca2+-blocking site that confers luminal pH and Ca2+ modulation on channel conductance. A luminal linker domain forms a fenestrated canopy atop the channel, providing several luminal ion passages to the pore and creating a negative electrostatic trap, with a preference for divalent cations, at the luminal entrance. The structure also reveals two equally distributed S4-S5 linker conformations in the closed channel, suggesting an S4-S5 linker-mediated PtdInsP2 gating mechanism among TRPML channels.


Evidence for essential histidine and cysteine residues in calcium/calmodulin-sensitive cyclic nucleotide phosphodiesterase.
Ca/calmodulin-sensitive cyclic nucleotide phosphodiesterase (CaM-PDE) is an important enzyme regulating cGMP levels and relaxation of vascular smooth muscle. This modification study was conducted mostly with bovine brain CaM-PDE to identify essential functional groups involved in catalysis. The effect of pH on Vmax/Km indicates two essential residues with pKa values of 6.4 and 8.2. Diethyl pyrocarbonate (DEP), a histidine-modifying agent, inhibits CaM-PDE with a second-order rate constant of 130 M-1 min-1 at pH 7.0 and 30 degrees C. Activity is restored by NH2OH. The pH dependence of inactivation reveals that the essential residue modified by DEP has an apparent pKa of 6.5. The difference spectrum of the intact and DEP-treated enzyme shows a maximum between 230 and 240 nm, suggesting formation of carbethoxy derivatives of histidine. The enzyme is also inactivated by N-ethylmaleimide (NEM) and 5,5'-dithiobis-(2-nitrobenzoic acid), both sulfhydryl-modifying agents, with the latter effect reversed by dithiothreitol, which suggests inactivation resulting from modification of cysteine residue(s). Partial inactivation of the enzyme by DEP or NEM results in an apparent decrease in the Vmax without a change in the Km or the extent of CaM stimulation. The rate of inactivation by DEP is greater in the presence than in the absence of Ca/CaM. A substrate analogue, Br-cGMP, and the competitive inhibitor 3-isobutyl-1-methylxanthine partially protect the enzyme against inactivation by DEP or NEM, suggesting that the modification of histidine and cysteine residues occurs at or near the active site. DEP also inactivated porcine brain CaM-PDE.(ABSTRACT TRUNCATED AT 250 WORDS)


A novel NRF2/ARE inhibitor gossypol induces cytotoxicity and sensitizes chemotherapy responses in chemo-refractory cancer cells.
NRF2/ARE signaling pathway is a principal regulator of cellular redox homoeostasis. The stress-induced transcription factor, NRF2, can shield cells from the oxidative damages via binding to the consensus antioxidant-responsive element (ARE) and driving several cyto-protective genes expression. Increasing evidence indicated that aberrant activation of NRF2 in malignant cells may support their survival through various pathways to detoxify chemotherapy drugs, attenuate drug-induced oxidative stress, or induce drug efflux, all of which are crucial in developing drug resistance. Accordingly, NRF2 is a potential drug target for improving the effectiveness of chemotherapy and to reverse drug resistance in cancer cells. A stable ARE-driven reporter human head and neck squamous cell carcinoma (HNSCC) cell line, HSC3-ARE9, was established and utilized to screen novel NRF2 inhibitors from a compound library. The cotton plant derived phenolic aldehyde-gossypol was selected for further analyses. The effects of gossypol in cancer cells were determined by western blotting, RT-qPCR, clonogenic assay, and cell viability assays. The gossypol-responsive gene expression levels were assessed in the Oncomine database. The effects of gossypol on conferring chemo-sensitization were evaluated in etoposide-resistant and cisplatin-resistant cancer cells. Our study is the first to identify that gossypol is effective to reduce both basal and NRF2 activator tert-butylhydroquinone (t-BHQ)-induced ARE-luciferase activity. Gossypol diminishes NRF2 protein stability and thereby leads to the suppression of NRF2/ARE pathway, which resulted in decreasing the expression levels of NRF2 downstream genes in both time- and dose-dependent manners. Inhibition of NRF2 by gossypol significantly decreases cell viabilities in human cancer cells. In addition, we find that gossypol re-sensitizes topoisomerase II poison treatment in etoposide-resistant cancer cells via suppression of NRF2/ABCC1 axis. Moreover, gossypol suppresses NRF2-mediated G6PD expression thereby leads to induce synthetic lethality with cisplatin not only in parental cancer cells but also in cisplatin-resistant cancer cells. These findings suggest that gossypol is a novel NRF2/ARE inhibitor, and can be a potential adjuvant chemotherapeutic agent for treatment of chemo-refractory tumor.


Combined transthoracic and transtracheal closure of large bronchopleural fistulae.
Postoperative central bronchopleural fistulae (BPF) are difficult to close using percutaneous or endoscopic techniques. We devised an alternative method to treat BPF using a combined transthoracic and transtracheal approach with the use of a multifilamented polypropylene (Prolene) mesh patch. Two patients with large, central BPF after thoracic surgery and lobar resection had minimally invasive BPF closure using a transtracheal approach with catheterization of the fistula and thoracoscopically guided Prolene mesh placement over the bronchial stump defect. This technique was adopted after conservative management and multiple endobronchial interventions had failed in both patients. One patient had closure of his BPF within one week and remains symptom-free one year after chest tube removal. The other patient had a BPF and chest tube for two years prior to our procedure. His BPF initially closed, but recannalized 2 weeks later. He subsequently had two thoracotomies and continues to suffer a BPF which remains externalized to his chest wall. Post-thoracotomy central BPF that is resistant to nonsurgical treatments can be closed with a combined thoracoscopic and transtracheal placement of a polypropylene patch. The success of this repair seems to depend on early intervention and aggressive sterilization of the pleural space.


Whole heart magnetization-prepared steady-state free precession coronary vein MRI.
To compare two coronary vein imaging techniques using whole-heart balanced steady-state free precession (SSFP) and a targeted double-oblique spoiled gradient-echo (GRE) sequences in combination with magnetization transfer (MT) preparation sequence for tissue contrast improvement. Nine healthy subjects were imaged with the proposed technique. The results are compared with optimized targeted MT prepared GRE acquisitions. Both quantitative and qualitative analyses were performed to evaluate each imaging method. Whole-heart images were successfully acquired with no visible image artifact in the vicinity of the coronary veins. The anatomical features and visual grading of both techniques were comparable. However, the targeted small slab acquisition of the left ventricular lateral wall was superior to whole-heart acquisition for visualization of relevant information for cardiac resynchronization therapy (CRT) lead implantation. We demonstrated the feasibility of whole-heart coronary vein MRI using a 3D MT-SSFP imaging sequence. A targeted acquisition along the lateral left ventricular wall is preferred for visualization of branches commonly used in CRT lead implantation.


Coupled dynamics of body mass and population growth in response to environmental change.
Environmental change has altered the phenology, morphological traits and population dynamics of many species. However, the links underlying these joint responses remain largely unknown owing to a paucity of long-term data and the lack of an appropriate analytical framework. Here we investigate the link between phenotypic and demographic responses to environmental change using a new methodology and a long-term (1976-2008) data set from a hibernating mammal (the yellow-bellied marmot) inhabiting a dynamic subalpine habitat. We demonstrate how earlier emergence from hibernation and earlier weaning of young has led to a longer growing season and larger body masses before hibernation. The resulting shift in both the phenotype and the relationship between phenotype and fitness components led to a decline in adult mortality, which in turn triggered an abrupt increase in population size in recent years. Direct and trait-mediated effects of environmental change made comparable contributions to the observed marked increase in population growth. Our results help explain how a shift in phenology can cause simultaneous phenotypic and demographic changes, and highlight the need for a theory integrating ecological and evolutionary dynamics in stochastic environments.


An essential role for Frizzled5 in neuronal survival in the parafascicular nucleus of the thalamus.
Frizzled5 (Fz5), a putative Wnt receptor, is expressed in the retina, hypothalamus, and the parafascicular nucleus (PFN) of the thalamus. By constructing Fz5 alleles in which beta-galactosidase replaces Fz5 or in which Cre-mediated recombination replaces Fz5 with alkaline phosphatase, we observe that Fz5 is required continuously and in a cell autonomous manner for the survival of adult PFN neurons, but is not required for proliferation, migration, or axonal growth and targeting of developing PFN neurons. A motor phenotype associated with loss of Fz5 establishes a role for the PFN in sensorimotor coordination. Transcripts coding for Wnt9b, the likely Fz5 ligand in vivo, and beta-catenin, a mediator of canonical Wnt signaling, are both downregulated in the Fz5(-/-) PFN, implying a positive feedback mechanism in which Wnt signaling is required to maintain the expression of Wnt signaling components. These data suggest that defects in Wnt-Frizzled signaling could be the cause of neuronal loss in degenerative CNS diseases.


The dynamics of reaction of Cl atoms with tetramethylsilane.
Rotational state distributions and state-selected CM-frame angular distributions were measured for HCl (v' = 0, j') products from the reaction of Cl-atoms with tetramethylsilane (TMS) under single collision conditions at a collision energy, E(coll), of 8.2 +/- 2.0 kcal mol(-1). The internal excitation of these products was very low with only 2% of the total energy available partitioned into HCl rotation. A transition state with a quasi-linear C-H-Cl moiety structure was computed and used to explain this finding. A backward peaking differential cross section was also reported together with a product translational energy (T') distribution with a maximum at T' approximately E(coll). This scattering behaviour is accounted for by reactions proceeding through a tight transition state on a highly skewed potential energy surface, which favours collisions at low impact parameters with a strong kinematic constraint on the internal excitation of the products. The large Arrhenius pre-exponential factor previously reported for this reaction is reconciled with the tight differential scattering observed in our study by considering the large size of the TMS molecule.


Where Sounds Occur Matters: Context Effects Influence Processing of Salient Vocalisations.
The social context in which a salient human vocalisation is heard shapes the affective information it conveys. However, few studies have investigated how visual contextual cues lead to differential processing of such vocalisations. The prefrontal cortex (PFC) is implicated in processing of contextual information and evaluation of saliency of vocalisations. Using functional Near-Infrared Spectroscopy (fNIRS), we investigated PFC responses of young adults (N = 18) to emotive infant and adult vocalisations while they passively viewed the scenes of two categories of environmental contexts: a domestic environment (DE) and an outdoors environment (OE). Compared to a home setting (DE) which is associated with a fixed mental representation (e.g., expect seeing a living room in a typical house), the outdoor setting (OE) is more variable and less predictable, thus might demand greater processing effort. From our previous study in Azhari et al. (2018) that employed the same experimental paradigm, the OE context was found to elicit greater physiological arousal compared to the DE context. Similarly, we hypothesised that greater PFC activation will be observed when salient vocalisations are paired with the OE compared to the DE condition. Our finding supported this hypothesis: the left rostrolateral PFC, an area of the brain that facilitates relational integration, exhibited greater activation in the OE than DE condition which suggests that greater cognitive resources are required to process outdoor situational information together with salient vocalisations. The result from this study bears relevance in deepening our understanding of how contextual information differentially modulates the processing of salient vocalisations.


Fibrogenesis imperfecta ossium.
The clinical features, investigation, treatment and outcome of two adults with fibrogenesis imperfecta ossium are described. In this rare acquired disorder of bone, normal lamellar collagen is replaced by structurally unsound collagen-deficient tissue, which leads to extreme bone fragility and ununited fractures. Transmission microscopy and SEM showed striking ultrastructural changes in bone structure and mineralisation. Both patients had monoclonal IgG paraproteins in the plasma and one excreted monoclonal lambda light chains in the urine. No abnormal plasma cells were found in the bone marrow and there was no evidence of amyloid deposition in the tissues. In both patients initial treatment with 1 alpha-hydroxycholecalciferol appeared to be ineffective, but in one, repeated courses of melphalan and corticosteroids over three years together with 1 alpha-hydroxycholecalciferol produced striking clinical and histological improvement. The findings in these and other patients strongly suggest that paraproteinaemia is an integral feature of fibrogenesis imperfecta ossium, and this needs further investigation.


Interference with Ca(2+) release activated Ca(2+) (CRAC) channel function delays T-cell arrest in vivo.
Entry of lymphocytes into secondary lymphoid organs (SLOs) involves intravascular arrest and intracellular calcium ion ([Ca(2+)]i) elevation. TCR activation triggers increased [Ca(2+)]i and can arrest T-cell motility in vitro. However, the requirement for [Ca(2+)]i elevation in arresting T cells in vivo has not been tested. Here, we have manipulated the Ca(2+) release-activated Ca(2+) (CRAC) channel pathway required for [Ca(2+)]i elevation in T cells through genetic deletion of stromal interaction molecule (STIM) 1 or by expression of a dominant-negative ORAI1 channel subunit (ORAI1-DN). Interestingly, the absence of CRAC did not interfere with homing of naïve CD4(+) T cells to SLOs and only moderately reduced crawling speeds in vivo. T cells expressing ORAI1-DN lacked TCR activation induced [Ca(2+)]i elevation, yet arrested motility similar to control T cells in vitro. In contrast, antigen-specific ORAI1-DN T cells had a twofold delayed onset of arrest following injection of OVA peptide in vivo. CRAC channel function is not required for homing to SLOs, but enhances spatiotemporal coordination of TCR signaling and motility arrest.


Forced expression of hepatocyte-specific fibroblast growth factor 21 delays initiation of chemically induced hepatocarcinogenesis.
Inappropriate fibroblast growth factor (FGF) signaling is involved in most tissue-specific pathologies including cancer. Previously we showed that inappropriate expression and chronic activity of FGF receptor (FGFR) 1 in hepatocytes accelerated diethylnitrosamine (DEN)-initiated hepatocarcinogenesis. Here we showed that although widely expressed FGF1 and FGF2 are frequently upregulated in hepatocellular carcinoma (HCC), germline deletion of both FGF1 and FGF2 had no effect on DEN-initiated hepatocarcinogenesis. Thus overexpression of FGF1 or FGF2 may be a consequence rather than contributor to hepatoma progression. FGF21 is the first of 22 homologues whose expression has been reported to be preferentially in the liver. We showed that similar to FGF1 and FGF2, FGF21 mRNA was upregulated in neoplastic and regenerating liver after partial hepatectomy (PH) and CCl4 administration. In situ hybridization analysis confirmed that in contrast to FGF1 and FGF2, expression of FGF21 mRNA was limited to hepatocytes. Forced overexpression of FGF21 in hepatocytes by gene targeting had no apparent impact on normal liver development and compensatory response to injury. Surprisingly, overexpression of FGF21 delayed the appearance of DEN-induced liver tumors. At 8 and 10 mo, only 10% and 30% of transgenic mice, respectively, developed adenomas compared to 50% (all adenomas) and 80% (60% adenoma/20% HCC) in the wild-type (WT) mice. However, the incidence and burden of HCC at 10 mo and later was equal in the FGF21 transgenic and WT mice. We propose that FGF21 may delay development of adenomas through activation of resident hepatocyte FGFR4 at early times, but counteracts the delay by acceleration of progression to HCC through interaction with ectopic FGFR1 once it appears in hepatoma cells. This indicates a dual function of FGF21 that may reflect changes in FGFR isotype during progression of differentiated hepatoma cells.


Origins in the Womb: Potential Role of the Physical Therapist in Modulating the Deleterious Effects of Obesity on Maternal and Offspring Health Through Movement Promotion and Prescription During Pregnancy.
Maternal obesity and associated metabolic disease contribute to adverse outcomes in women and their offspring, and many of these outcomes have significant acute and chronic implications for both mother and neonate. Targeted movement (ie, physical activity or exercise training) during pregnancy has been shown to be safe and effective for improving many of these outcomes in women at a healthy weight and women who are obese. However, movement prescription and advice during pregnancy are often not addressed by health care providers; this situation creates a unique opportunity for physical therapists to use their expertise in movement with patients who are pregnant. The objective of this article is to briefly review the adverse maternal and neonatal outcomes associated with maternal obesity, the benefits of intentional maternal movement during pregnancy for women who are obese, the evidence-based guidelines for prescribing intentional movement during pregnancy for women who are obese, and the potential for physical therapists to become the driving force behind a necessary increase in movement levels in women who are pregnant. Physical therapists can play a significant role in encouraging movement in women who are healthy and women who have metabolic challenges during pregnancy and thus assist in combating the vicious cycle of obesity by improving maternal and offspring health.


Angina and Future Cardiovascular Events in Stable Patients With Coronary Artery Disease: Insights From the Reduction of Atherothrombosis for Continued Health (REACH) Registry.
The extent to which angina is associated with future cardiovascular events in patients with coronary artery disease has long been debated. Included were outpatients with established coronary artery disease who were enrolled in the REACH registry and were followed for 4 years. Angina at baseline was defined as necessitating episodic or permanent antianginal treatment. The primary end point was the composite of cardiovascular death, myocardial infarction, or stroke. Secondary end points included heart failure, cardiovascular hospitalizations, and coronary revascularization. The independent association between angina and first/total events was examined using Cox and logistic regression models. Out of 26 159 patients with established coronary artery disease, 13 619 (52%) had angina at baseline. Compared with patients without angina, patients with angina were more likely to be older, female, and had more heart failure and polyvascular disease (P<0.001 for each). Compared with patients without angina, patients with angina had higher rates of first primary end-point event (14.2% versus 16.3%, unadjusted hazard ratio 1.19, CI 1.11-1.27, P<0.001; adjusted hazard ratio 1.06, CI 0.99-1.14, P=0.11), and total primary end-point events (adjusted risk ratio 1.08, CI 1.01-1.16, P=0.03). Patients with angina were at increased risk for heart failure (adjusted odds ratio 1.17, CI 1.06-1.28, P=0.002), cardiovascular hospitalizations (adjusted odds ratio 1.29, CI 1.21-1.38, P<0.001), and coronary revascularization (adjusted odds ratio 1.23, CI 1.13-1.34, P<0.001). Patients with stable coronary artery disease and angina have higher rates of future cardiovascular events compared with patients without angina. After adjustment, angina was only weakly associated with cardiovascular death, myocardial infarction, or stroke, but significantly associated with heart failure, cardiovascular hospitalization, and coronary revascularization.


Investigating the Causal Relationship of C-Reactive Protein with 32 Complex Somatic and Psychiatric Outcomes: A Large-Scale Cross-Consortium Mendelian Randomization Study.
C-reactive protein (CRP) is associated with immune, cardiometabolic, and psychiatric traits and diseases. Yet it is inconclusive whether these associations are causal. We performed Mendelian randomization (MR) analyses using two genetic risk scores (GRSs) as instrumental variables (IVs). The first GRS consisted of four single nucleotide polymorphisms (SNPs) in the CRP gene (GRSCRP), and the second consisted of 18 SNPs that were significantly associated with CRP levels in the largest genome-wide association study (GWAS) to date (GRSGWAS). To optimize power, we used summary statistics from GWAS consortia and tested the association of these two GRSs with 32 complex somatic and psychiatric outcomes, with up to 123,865 participants per outcome from populations of European ancestry. We performed heterogeneity tests to disentangle the pleiotropic effect of IVs. A Bonferroni-corrected significance level of less than 0.0016 was considered statistically significant. An observed p-value equal to or less than 0.05 was considered nominally significant evidence for a potential causal association, yet to be confirmed. The strengths (F-statistics) of the IVs were 31.92-3,761.29 and 82.32-9,403.21 for GRSCRP and GRSGWAS, respectively. CRP GRSGWAS showed a statistically significant protective relationship of a 10% genetically elevated CRP level with the risk of schizophrenia (odds ratio [OR] 0.86 [95% CI 0.79-0.94]; p < 0.001). We validated this finding with individual-level genotype data from the schizophrenia GWAS (OR 0.96 [95% CI 0.94-0.98]; p < 1.72 × 10-6). Further, we found that a standardized CRP polygenic risk score (CRPPRS) at p-value thresholds of 1 × 10-4, 0.001, 0.01, 0.05, and 0.1 using individual-level data also showed a protective effect (OR < 1.00) against schizophrenia; the first CRPPRS (built of SNPs with p < 1 × 10-4) showed a statistically significant (p < 2.45 × 10-4) protective effect with an OR of 0.97 (95% CI 0.95-0.99). The CRP GRSGWAS showed that a 10% increase in genetically determined CRP level was significantly associated with coronary artery disease (OR 0.88 [95% CI 0.84-0.94]; p < 2.4 × 10-5) and was nominally associated with the risk of inflammatory bowel disease (OR 0.85 [95% CI 0.74-0.98]; p < 0.03), Crohn disease (OR 0.81 [95% CI 0.70-0.94]; p < 0.005), psoriatic arthritis (OR 1.36 [95% CI 1.00-1.84]; p < 0.049), knee osteoarthritis (OR 1.17 [95% CI 1.01-1.36]; p < 0.04), and bipolar disorder (OR 1.21 [95% CI 1.05-1.40]; p < 0.007) and with an increase of 0.72 (95% CI 0.11-1.34; p < 0.02) mm Hg in systolic blood pressure, 0.45 (95% CI 0.06-0.84; p < 0.02) mm Hg in diastolic blood pressure, 0.01 ml/min/1.73 m2 (95% CI 0.003-0.02; p < 0.005) in estimated glomerular filtration rate from serum creatinine, 0.01 g/dl (95% CI 0.0004-0.02; p < 0.04) in serum albumin level, and 0.03 g/dl (95% CI 0.008-0.05; p < 0.009) in serum protein level. However, after adjustment for heterogeneity, neither GRS showed a significant effect of CRP level (at p < 0.0016) on any of these outcomes, including coronary artery disease, nor on the other 20 complex outcomes studied. Our study has two potential limitations: the limited variance explained by our genetic instruments modeling CRP levels in blood and the unobserved bias introduced by the use of summary statistics in our MR analyses. Genetically elevated CRP levels showed a significant potentially protective causal relationship with risk of schizophrenia. We observed nominal evidence at an observed p < 0.05 using either GRSCRP or GRSGWAS-with persistence after correction for heterogeneity-for a causal relationship of elevated CRP levels with psoriatic osteoarthritis, rheumatoid arthritis, knee osteoarthritis, systolic blood pressure, diastolic blood pressure, serum albumin, and bipolar disorder. These associations remain yet to be confirmed. We cannot verify any causal effect of CRP level on any of the other common somatic and neuropsychiatric outcomes investigated in the present study. This implies that interventions that lower CRP level are unlikely to result in decreased risk for the majority of common complex outcomes.


Health education for musicians in the UK: a qualitative evaluation.
Musical training in higher education music institutions (e.g. conservatoires) has been associated with health-related issues among musicians. The Health Promotion in Schools of Music project in the USA and the Healthy Conservatoires project in the UK have therefore recommended health promotion at conservatoires. Few health education courses have been evaluated to date, however. A 5-month health education programme for first-year undergraduate students at a British conservatoire was introduced as part of the core curriculum in September 2016. The programme, which involved both lectures and seminars, was evaluated using quantitative and qualitative approaches. This article reports only the qualitative evaluation. Twenty semi-structured individual interviews were conducted either face-to-face or via Skype in April 2017. The data were transcribed verbatim and analysed thematically. Five themes were identified: (i) the programme as a catalyst for engagement with health; (ii) behavioural changes; (iii) barriers to engaging with the programme material and initiating changes; (iv) suggestions for improvement; and (v) misinformation. Generally, participants viewed the programme as relevant and informative, particularly appreciating the intimate nature of the seminars. They reported that the programme helped them take a broader perspective on musicianship and that they would welcome sessions that are more practical than theoretical. They also reported instances of change in their behaviours relating to both lifestyle and management of music practice. In conclusion, undergraduate music students viewed this health education programme positively. Their feedback illustrates the complex nature of health promotion in the conservatoire setting.


Practitioner based quality improvement: a review of the Royal College of Nursing's dynamic standard setting system.
To explore and describe the implementation of the Royal College of Nursing's approach to audit--the dynamic standard setting system--within the current context of health care, in particular to focus on how the system has developed since its inception in the 1980s as a method for uniprofessional and multiprofessional audit. Qualitative design with semistructure interviews and field visits. 28 sites throughout the United Kingdom that use the dynamic standard setting system. Quality and audit coordinators with a responsibility for implementing the system; clinical staff who practice the system. Experiences of the dynamic standard setting system, including reasons for selection, methods of implementation, and observed outcomes. Issues relating to four themes emerged from the data: practical experiences of the system as a method for improving patient care; issues of facilitation and training; strategic issues of implementation; and the use of the system as a method for multiprofessional audit. The development of clinical practice was described as a major benefit of the system and evidence of improved patient care was apparent. However, difficulties were experienced in motivating staff and finding time for audit, which in part related to the current format of the system and the level of training and support available for clinical staff. Diverse experiences were reported in the extent to which the system had been integrated at a strategic level of quality improvement and its successful application to multiprofessional clinical audit. The Royal College of Nursing's dynamic standard setting system can successfully be used as a method for clinical audit at both a uniprofessional and multiprofessional level. However, to capitalise on the strengths of the system, several issues need to be considered further. These include modifications to the system itself, as well as a more strategic focus on resources and support for audit, better integration of quality initiatives in health care, and a continuing focus on ways to achieve true multiprofessional collaboration and involvement of patients in clinical audit.


Noise and the evolution of neural network modularity.
We study the selective advantage of modularity in artificially evolved networks. Modularity abounds in complex systems in the real world. However, experimental evidence for the selective advantage of network modularity has been elusive unless it has been supported or mandated by the genetic representation. The evolutionary origin of modularity is thus still debated: whether networks are modular because of the process that created them, or the process has evolved to produce modular networks. It is commonly argued that network modularity is beneficial under noisy conditions, but experimental support for this is still very limited. In this article, we evolve nonlinear artificial neural network classifiers for a binary classification task with a modular structure. When noise is added to the edge weights of the networks, modular network topologies evolve, even without representational support.


Native Escherichia coli OmpF porin surfaces probed by atomic force microscopy.
Topographs of two-dimensional porin OmpF crystals reconstituted in the presence of lipids were recorded in solution by atomic force microscopy (AFM) to a lateral resolution of 10 angstroms and a vertical resolution of 1 angstrom. Protein-protein interactions were demonstrated on the basis of the AFM results and earlier crystallographic findings. To assess protein-lipid interactions, the bilayer was modeled with kinked lipids by fitting the head groups to contours determined with AFM. Finally, two conformations of the extracellular porin surface were detected at forces of 0.1 nanonewton, demonstrating the potential of AFM to monitor conformational changes with high resolution.


Positive correlation between histological tumor involvement and generation of tumor cell colonies in agarose in specimens taken directly from patients with small-cell carcinoma of the lung.
Twenty-seven specimens for in vitro agarose clonogenicity testing were obtained from 25 patients with small-cell carcinoma of the lung (SCCL). The specimens were obtained from bone marrows, pleural effusions, lymph nodes, and liver biopsies. Colony formation was seen in 14 of 15 specimens that were histologically involved with SCCL, but no colony growth was seen in the 12 patient specimens without histocytopathological evidence of SCCL, including seven bone marrow specimens. Cytological examination of the agarose colonies confirmed their SCCL origin. Colonies reached sizes of 50 to 1000 cells in 7 to 10 days, indicating an in vitro doubling time of less than 24 hr, remarkably shorter than the population doubling times measured in patients. None of the 100 clones picked from these specimens demonstrated the ability to continuously replicate in vitro. These results show an excellent correlation between agarose colony formation and histological tumor involvement and a more rapid in vitro doubling time than that seen in vivo and demonstrate that standard tissue culture conditions do not allow demonstration of a self-renewing stem cell in fresh tumor specimens of SCCL.


Determination of low urinary 8-hydroxy-2`-deoxyguanosine excretion with capillary electrophoresis and molecularly imprinted monolith solid phase microextraction.
As a sensitive biomarker, the level of 8-hydroxy-2`-deoxyguanosine (8-OHdG) indicates the extent of DNA oxidative damage of subject. However its determination in human urine is confounded by trace content and a complex matrix. In this study, a new approach of solid phase microextraction (SPME) coupled to capillary electrophoresis and electrochemical detection was developed with molecularly imprinted monolithic column as extraction sorbent. The tailor made monolith exhibited high extraction efficiency with the enrichment factor 73 for 8-OHdG owing to its special porous structure and inherent selectivity. Under optimal conditions, appreciable sensitivity had been achieved with a limit of detection of 2.61 nmol/L (signal/noise=3) and a limit of quantification of 8.63 nmol/L (signal/noise=10), respectively. As an alternative of precise 8-OHdG determination, wide range linearity had been achieved with 0.01-1.50 μmol/L and RSDs with 3.70% for migration time and 4.80% for peak current (10-1000 nmol/L). 106 samples with relative low extents of DNA oxidative damage were collected and analyzed which including coke plant workers, traffic policemen, taxi drivers and healthy students. Elevated urinary 8-OHdG excretions of subjects may correlate to working condition, occupational exposure, or lifestyle.


Mining twitter: a source for psychological wisdom of the crowds.
Over the last few years, microblogging has gained prominence as a form of personal broadcasting media where information and opinion are mixed together without an established order, usually tightly linked with current reality. Location awareness and promptness provide researchers using the Internet with the opportunity to create "psychological landscapes"--that is, to detect differences and changes in voiced (twittered) emotions, cognitions, and behaviors. In our article, we present iScience Maps, a free Web service for researchers, available from http://maps.iscience.deusto.es/ and http://tweetminer.eu/ . Technologically, the service is based on Twitter's streaming and search application programming interfaces (APIs), accessed through several PHP libraries, and a JavaScript frontend. This service allows researchers to assess via Twitter the effect of specific events in different places as they are happening and to make comparisons between cities, regions, or countries regarding psychological states and their evolution in the course of an event. In a step-by-step example, it is shown how to replicate a study on affective and personality characteristics inferred from first names (Mehrabian & Piercy, Personality and Social Psychology Bulletin, 19, 755-758 1993) by mining Twitter data with iScience Maps.Results from the original study are replicated in both world regions we tested (the western U.S. and the U.K./Ireland); we also discover base rate of names to be a confound that needs to be controlled for in future research.


Metformin Use Is Associated With Reduced Mortality in a Diverse Population With COVID-19 and Diabetes.
Coronavirus disease-2019 (COVID-19) is a growing pandemic with an increasing death toll that has been linked to various comorbidities as well as racial disparity. However, the specific characteristics of these at-risk populations are still not known and approaches to lower mortality are lacking. We conducted a retrospective electronic health record data analysis of 25,326 subjects tested for COVID-19 between 2/25/20 and 6/22/20 at the University of Alabama at Birmingham Hospital, a tertiary health care center in the racially diverse Southern U.S. The primary outcome was mortality in COVID-19-positive subjects and the association with subject characteristics and comorbidities was analyzed using simple and multiple linear logistic regression. The odds ratio of contracting COVID-19 was disproportionately high in Blacks/African-Americans (OR 2.6; 95% CI 2.19-3.10; p<0.0001) and in subjects with obesity (OR 1.93; 95% CI 1.64-2.28; p<0.0001), hypertension (OR 2.46; 95% CI 2.07-2.93; p<0.0001), and diabetes (OR 2.11; 95% CI 1.78-2.48; p<0.0001). Diabetes was also associated with a dramatic increase in mortality (OR 3.62; 95% CI 2.11-6.2; p<0.0001) and emerged as an independent risk factor in this diverse population even after correcting for age, race, sex, obesity, and hypertension. Interestingly, we found that metformin treatment prior to diagnosis of COVID-19 was independently associated with a significant reduction in mortality in subjects with diabetes and COVID-19 (OR 0.33; 95% CI 0.13-0.84; p=0.0210). Thus, these results suggest that while diabetes is an independent risk factor for COVID-19-related mortality, this risk is dramatically reduced in subjects taking metformin prior to diagnosis of COVID-19, raising the possibility that metformin may provide a protective approach in this high risk population.


The influence of latent toxoplasmosis on women's reproductive function: four cross-sectional studies.
Several studies have investigated the association between infection with Toxoplasma gondii (Nicolle et Manceaux, 1908), pregnancy and fertility, but the results of studies focused on the fertility are rather ambiguous. Here we report results of four new cross-sectional studies. The studies were performed in the General University Hospital, Prague (study A with n = 1 165, and study C with n = 317), in private clinics of the Centre of Reproductive Medicine, Prague (study B with n = 1 016), and in a population of Czech and Slovak volunteers from the Facebook page 'Guinea Pigs' willing to participate in various basic science studies (study D with n = 524). In studies A and B, the clinical records were used to assess the fertility problems, whereas in studies C and D, the women were asked to rate their fertility problems using a six-point scale. Pregnant T. gondii-infected women were older than T. gondii-free women (study A: 33.1 vs 31.2, P < 0.001; study B: 30.6 vs 29.6, P = 0.012) and more often used assisted reproductive technology to conceive (study A: 17.2% vs 12.4%, P = 0.041; study B: 13.4% vs 9.2%, P = 0.317). Pregnant T. gondii-infected primiparous women were older than T. gondii-free primiparas (study A: 31.1 vs 29.5, P < 0.001; study B: 29.7 vs 28.9, P = 0.064) and more often used assisted reproductive technology to conceive (study A: 24.7% vs 14.4%, P = 0.010; study B: 15.9% vs 15.5%, P = 0.888). T. gondii-infected women reported to take a longer time to conceive than T. gondii-free women (P = 0.015). They also claimed to have more fertility problems than T. gondii-free women (P < 0.0001). Our results suggest that 'asymptomatic' latent toxoplasmosis could be a more serious source of fertility problems and health-associated burden than more severe but far rarer congenital toxoplasmosis.


The importance of age and smoking in evaluating adverse cytogenetic effects of exposure to environmental agents.
Fluorescence in situ hybridization with chromosome-specific composite DNA probes (chromosome painting) is a reliable and efficient method for detecting structural chromosome aberrations. Painting is now being used to quantify chromosome damage in many human populations. In one such study we evaluated 91 unexposed people ranging in age from birth (cord bloods) to 79. We established a baseline frequency of stable aberrations that showed a highly significant curvilinear increase with age (p < 0.00001) that accounted for 70% of the variance among donors. The magnitude of this effect illustrates the importance of understanding the cytogenetic changes that occur with age, which is particularly important for quantifying the effects of prior adverse environmental, occupational, or accidental exposure. In this paper we use the data obtained in our previous study to characterize the distribution of stable aberrations by age and pack-years of cigarette smoking. We also provide estimates of the number of cell equivalents that need to be scored to detect a given increase in aberrations above the background level surveyed in this population.


Impact of left ventricular ejection fraction on clinical outcomes over five years after infarct-related coronary artery recanalization (from the Occluded Artery Trial [OAT]).
In the Occluded Artery Trial (OAT), percutaneous coronary intervention (PCI) of an infarct-related artery on days 3 to 28 after acute myocardial infarction was of no benefit compared to medical therapy alone. The present analysis was conducted to determine whether PCI might provide benefit to the subgroup of higher risk patients with a depressed ejection fraction (EF). Of 2,185 analyzed patients (age 58.6 +/- 11.0 years) with infarct-related artery occlusion on days 3 to 28 after acute myocardial infarction in the Occluded Artery Trial, 1,094 were assigned to PCI and 1,091 to medical therapy. The primary end point was a composite of death, reinfarction, and New York Heart Association class IV heart failure. The outcomes were analyzed by EF (first tertile, EF < or =44%, vs second and third tertiles combined, EF >44%). Interaction of the treatment effect with EF on the study outcomes were examined using the Cox survival model. The 5-year rates of the primary end point (death, reinfarction, or New York Heart Association class IV heart failure) were not different in either subgroup (PCI vs medical therapy, hazard ratio 1.25, 99% confidence interval 0.83 to 1.88, for EF < or =44%; hazard ratio 0.98, 99% confidence interval 0.64 to 1.50, for EF >44%). However, in patients with an EF >44%, PCI reduced the rate of subsequent revascularization (p = 0.004, interaction p = 0.05). In conclusion, optimal medical therapy remains the overall treatment of choice for stable patients with a persistent total occlusion of the infarct-related artery after acute myocardial infarction, irrespective of the baseline EF. In patients with normal or moderately impaired left ventricular contractility, PCI reduced the need for subsequent revascularization but did not otherwise improve outcomes.


Brazilian Child Protection Professionals' Resilient Behavior during the COVID-19 Pandemic.
The COVID-19 pandemic deeply affected child protection professionals. One potential area of concern is whether and how the pandemic has dampened these individuals' ability to engage in the resilient practices that are so vital to their wellbeing. Within the unique and understudied context of a developing economy facing the strain of an international pandemic, this study sought to expand our theoretical understanding of the individual and socio-ecological predictors of whether child protective services professionals engage in resilient behaviors. Three hundred and nine professionals working in child protection related fields (e.g., psychologists, social workers, professors, pediatricians, nurses, and other clinicians). Participants were surveyed as to their demographics, current work conditions, their engagement in resilient behaviors, and potential individual and socio-ecological predictors of those behaviors. Both job support for and individuals' beliefs of the importance of resilient behaviors predicted their engagement in such behaviors. Child protection professionals' resilience must be fostered by socio-ecological contexts, such as their workplace and employers, and additional supports are needed during the trying times of the COVID-19 pandemic.


A review of 40 years of enteric antimicrobial resistance research in Eastern Africa: what can be done better?
The emergence and persistence of antimicrobial resistance is driven by varied factors including the indiscriminate use of antibiotics and variable drug efficacy and presents a major threat to the control of infectious diseases. Despite the high burden of disease in sub-Saharan Africa and the potential health and economic consequences, the level of research on antimicrobial resistance in the region remains unknown. Little data exists to quantify the contribution of different factors to the current levels of antimicrobial resistance. To identify the factors that contribute most to the emergence, amplification, persistence and dissemination of antimicrobial resistance in humans and animals, we used the PRISMA 2009 guidelines to conduct a systematic review of studies on antibiotic-resistant enteric bacteria in Eastern Africa. We searched PubMed and Google Scholar databases and identified 2,155 probable articles, of which 89 studies on humans and 28 on animals remained after full-text review. These were articles from Kenya, Tanzania, Uganda, Ethiopia, Rwanda and Burundi, published between 1974 and 2013, that reported resistance in Salmonella, Shigella, Escherichia coli and Vibrio sp. The majority (98%) of human studies were based on hospital- (rather than community-wide) sampling and although they report high levels of antimicrobial resistance in the region, study design and methodological differences preclude conclusions about the magnitude and trends of antimicrobial resistance. To remedy this, we discuss and propose minimum reporting guidelines for the level of detail that should be explicitly provided for antimicrobial resistance study designs, testing of samples and reporting of results that would permit comparative inferences and enable meta-analyses. Further, we advocate for increased focus on community- rather than hospital-based sampling to provide a better indication of population-wide trends in antimicrobial resistance. This approach, together with the establishment of a robust regional surveillance network, should over time build a pool of evidence-based data useful for policy decisions and interventions aimed at controlling antimicrobial resistance.


Change in the chemical composition of infalling gas forming a disk around a protostar.
IRAS 04368+2557 is a solar-type (low-mass) protostar embedded in a protostellar core (L1527) in the Taurus molecular cloud, which is only 140 parsecs away from Earth, making it the closest large star-forming region. The protostellar envelope has a flattened shape with a diameter of a thousand astronomical units (1 AU is the distance from Earth to the Sun), and is infalling and rotating. It also has a protostellar disk with a radius of 90 AU (ref. 6), from which a planetary system is expected to form. The interstellar gas, mainly consisting of hydrogen molecules, undergoes a change in density of about three orders of magnitude as it collapses from the envelope into the disk, while being heated from 10 kelvin to over 100 kelvin in the mid-plane, but it has hitherto not been possible to explore changes in chemical composition associated with this collapse. Here we report that the unsaturated hydrocarbon molecule cyclic-C3H2 resides in the infalling rotating envelope, whereas sulphur monoxide (SO) is enhanced in the transition zone at the radius of the centrifugal barrier (100 ± 20 AU), which is the radius at which the kinetic energy of the infalling gas is converted to rotational energy. Such a drastic change in chemistry at the centrifugal barrier was not anticipated, but is probably caused by the discontinuous infalling motion at the centrifugal barrier and local heating processes there.


Heritability of C-reactive protein and association with apolipoprotein E genotypes in Japanese Americans.
Numerous studies have demonstrated that increased C-reactive protein (CRP) levels predict coronary heart disease, stroke, peripheral vascular disease, and diabetes, and are associated with features of the metabolic syndrome. Only three previous studies have investigated the heritability of CRP levels, primarily in samples of Caucasian families. The purpose of the present study was to estimate the magnitude of genetic influences on CRP levels, and to examine potential associations between variation in the APOE gene and CRP levels, using a sample of 562 individual Japanese Americans from 68 extended kindreds. In general, correlation coefficients between first-degree relatives for CRP were approximately 0.2, and spouse correlations did not differ from zero, consistent with genetic influences. Heritability estimates were approximately 0.3 (p < 0.01), even with adjustment for factors known to influence CRP levels. A significant relationship was seen between unadjusted CRP levels and APOE genotypes (p = 0.02), with the highest mean CRP level among epsilon2 carriers (1.20 mg/L), and nearly the same mean levels among epsilon3/epsilon3 subjects and epsilon4 carriers (0.72 and 0.74 mg/L, respectively). However, this relationship was diminished with adjustment for covariates (p = 0.07). These results demonstrate the presence of both genetic and environmental effects on CRP levels among Asian Americans, and additional studies are needed to determine if the APOE gene contributes to these genetic influences.


From research to national expansion: 20 years' experience of community-based management of childhood pneumonia in Nepal.
Pneumonia is a leading cause of mortality of children aged under five in Nepal. Research conducted by John Snow Inc. in the 1980s determined that pneumonia case management by community-based workers decreased under-five mortality by 28%. Female community health volunteers were selected as the national cadre to manage childhood pneumonia at community level using oral antibiotics. A technical working group composed of government officials, local experts and donor partners embarked on a process to develop a strategy to pilot the approach and expand it nationally. High under-five mortality rates, low access to peripheral health facilities and severe constraints in human resources led Nepal's Ministry of Health to test this innovative approach. Community-based management of pneumonia doubled the total number of cases treated compared with districts with facility-based treatment only. Over half of the cases were treated by the female community health volunteers. The programme was phased in over 14 years and now 69% of Nepal's under-five population has access to pneumonia treatment. Community-based management of pneumonia provides a medium-term solution to address a leading cause of child mortality while the efforts continue to strengthen and extend the reach of facility-based care. Trained community health workers can significantly increase the number of pneumonia cases receiving correct case management in resource-constrained settings, with appropriate health systems' support for logistics, supervision and monitoring. Community-based management of pneumonia can be scaled up and provides an effective approach to reducing child deaths in countries faced with insufficient human resources for health.


Visual traffic jam analysis based on trajectory data.
In this work, we present an interactive system for visual analysis of urban traffic congestion based on GPS trajectories. For these trajectories we develop strategies to extract and derive traffic jam information. After cleaning the trajectories, they are matched to a road network. Subsequently, traffic speed on each road segment is computed and traffic jam events are automatically detected. Spatially and temporally related events are concatenated in, so-called, traffic jam propagation graphs. These graphs form a high-level description of a traffic jam and its propagation in time and space. Our system provides multiple views for visually exploring and analyzing the traffic condition of a large city as a whole, on the level of propagation graphs, and on road segment level. Case studies with 24 days of taxi GPS trajectories collected in Beijing demonstrate the effectiveness of our system.


Sample Preparation and Warping Accuracy for Correlative Multimodal Imaging in the Mouse Olfactory Bulb Using 2-Photon, Synchrotron X-Ray and Volume Electron Microscopy.
Integrating physiology with structural insights of the same neuronal circuit provides a unique approach to understanding how the mammalian brain computes information. However, combining the techniques that provide both streams of data represents an experimental challenge. When studying glomerular column circuits in the mouse olfactory bulb, this approach involves e.g., recording the neuronal activity with in vivo 2-photon (2P) calcium imaging, retrieving the circuit structure with synchrotron X-ray computed tomography with propagation-based phase contrast (SXRT) and/or serial block-face scanning electron microscopy (SBEM) and correlating these datasets. Sample preparation and dataset correlation are two key bottlenecks in this correlative workflow. Here, we first quantify the occurrence of different artefacts when staining tissue slices with heavy metals to generate X-ray or electron contrast. We report improvements in the staining procedure, ultimately achieving perfect staining in ∼67% of the 0.6 mm thick olfactory bulb slices that were previously imaged in vivo with 2P. Secondly, we characterise the accuracy of the spatial correlation between functional and structural datasets. We demonstrate that direct, single-cell precise correlation between in vivo 2P and SXRT tissue volumes is possible and as reliable as correlating between 2P and SBEM. Altogether, these results pave the way for experiments that require retrieving physiology, circuit structure and synaptic signatures in targeted regions. These correlative function-structure studies will bring a more complete understanding of mammalian olfactory processing across spatial scales and time.


Cutting edge: hypoxia-inducible factor 1alpha and its activation-inducible short isoform I.1 negatively regulate functions of CD4+ and CD8+ T lymphocytes.
To evaluate the role of hypoxia-inducible factor 1alpha (HIF-1alpha) and its TCR activation-inducible short isoform I.1 in T cell functions, we genetically engineered unique mice with: 1) knockout of I.1 isoform of HIF-1alpha; 2) T cell-targeted HIF-1alpha knockdown; and 3) chimeric mice with HIF-1alpha gene deletion in T and B lymphocytes. In all three types of mice, the HIF-1alpha-deficient T lymphocytes, which were TCR-activated in vitro, produced more proinflammatory cytokines compared with HIF-1alpha-expressing control T cells. Surprisingly, deletion of the I.1 isoform, which represents < 30% of total HIF-1alpha mRNA in activated T cells, was sufficient to markedly enhance TCR-triggered cytokine secretion. These data suggest that HIF-1alpha not only plays a critical role in oxygen homeostasis but also may serve as a negative regulator of T cells.


Heterogeneous surface expression of EspA translocon filaments by Escherichia coli O157:H7 is controlled at the posttranscriptional level.
Type III secretion systems of enteric bacteria enable translocation of effector proteins into host cells. Secreted proteins of verotoxigenic Escherichia coli O157 strains include components of a translocation apparatus, EspA, -B, and -D, as well as "effectors" such as the translocated intimin receptor (Tir) and the mitochondrion-associated protein (Map). This research has investigated the regulation of LEE4 translocon proteins, in particular EspA. EspA filaments could not be detected on the bacterial cell surface when E. coli O157:H7 was cultured in M9 minimal medium but were expressed from only a proportion of the bacterial population when cultured in minimal essential medium modified with 25 mM HEPES. The highest proportions of EspA-filamented bacteria were detected in late exponential phase, after which filaments were lost rapidly from the bacterial cell surface. Our previous research had shown that human and bovine E. coli O157:H7 strains exhibit marked differences in EspD secretion levels. Here it is demonstrated that the proportion of the bacterial population expressing EspA filaments was associated with the level of EspD secretion. The ability of individual bacteria to express EspA filaments was not controlled at the level of LEE1-4 operon transcription, as demonstrated by using both beta-galactosidase and green fluorescent protein (GFP) promoter fusions. All bacteria, whether expressing EspA filaments or not, showed equivalent levels of GFP expression when LEE1-4 translational fusions were used. Despite this, the LEE4-espADB mRNA was more abundant from populations with a high proportion of nonsecreting bacteria (low secretors) than from populations with a high proportion of secreting and therefore filamented bacteria (high secretors). This research demonstrates that while specific environmental conditions are required to induce LEE1-4 expression, a further checkpoint exists before EspA filaments are produced on the bacterial surface and secretion of effector proteins occurs. This checkpoint in E. coli O157:H7 translocon expression is controlled by a posttranscriptional mechanism acting on LEE4-espADB mRNA. The heterogeneity in EspA filamentation could arise from phase-variable expression of regulators that control this posttranscriptional mechanism.


Evalution of Femicide Cases Committed Between the Years 1996-2005 in Antalya.
Criminal death incidence of women varies between developed and developing countries, and it has become a common public problem in almost every country. We do not have any data about femicide cases published in Turkey until today. In our study, we tried to call attention to femicide cases. In our study, we evaluated 141 cases of female homicides with an interval of 10 years between January 1996 and May 2005, retrospectively. Data retrieved were statistically evaluated using chi-square test. Most of the cases were between 21 and 35 years of age. Spouse murders are usually witnessed during the process of divorce. Establishment of institutions which will ensure the security of women during this period is of paramount importance. If we protect women during divorce and separation proceedings, we can prevent femicide cases. We also think that laws may be strengthened for protecting women who are vulnerable at such times.


Moving to productivity: The benefits of healthy buildings.
Health is a critical factor for the generation of value by workers. Companies bear substantial costs associated with absenteeism and presenteeism among their employees. This study investigates the impact of the environmental conditions in the workplace on the health and job satisfaction of employees, as core factors of productivity. We provide evidence based on a natural experiment, in which 70% of the workforce of a municipality in the Netherlands was relocated to a building with a design focused on sustainability and health and well-being. We construct a longitudinal dataset based on individual surveys of the entire municipality workforce and include measures before and after the move. The estimation results show a significant improvement in the perceived environmental conditions, as well as in the health and well-being of the relocated workers, measured by the drop in incidence of sick building syndrome symptoms. Results are heterogeneous based on age: older groups of employees enjoy larger health impacts. The relocation effects remain persistent in the medium term (two years after the moving date). Importantly, a mediation analysis suggests that the achieved improvements in health and well-being lead to significantly enhanced job satisfaction and a 2% reduction in the prevalence of sick leave.


Subaqueous explosive eruption and welding of pyroclastic deposits.
Silicic tuffs infilling an ancient submarine caldera, at Mineral King in California, show microscopic fabrics indicative of welding of glass shards and pumice at temperatures >500 degrees C. The occurrence indicates that subaqueous explosive eruption and emplacement of pyroclastic materials can occur without substantial admixture of the ambient water, which would cause chilling. Intracaldera progressive aggradation of pumice and ash from a thick, fast-moving pyroclastic flow occurred during a short-lived explosive eruption of approximately 26 cubic kilometers of magma in water >/=150 meters deep. The thickness, high velocity, and abundant fine material of the erupted gas-solids mixture prevented substantial incorporation of ambient water into the flow. Stripping of pyroclasts from upper surfaces of subaqueous pyroclastic flows in general, both above the vent and along any flow path, may be the main process giving rise to buoyant-convective subaqueous eruption columns and attendant fallout deposits.


What essential medicines for children are on the shelf?
To document the inclusion of key medicines for children in national essential medicines lists (EMLs) and standard treatment guidelines, and to assess the availability and cost of these medicines in 14 countries in central Africa. Surveys were conducted in 12 public and private sector medicine outlets in each country's capital city. Data were collected on medicine availability on the survey day and on the cost to the patient of the lowest priced medicine in stock. The proportion of survey medicines in national EMLs ranged from 50% to 90%. In only three countries were more than 50% of such medicines available from central medical stores (range: 15-75%). Availability in nongovernmental organization stores was not consistently better (range: 10-65%) but tended to be higher in teaching hospitals, although the range was similar (15-70%). District hospitals (range: 10-80%) had slightly better availability than teaching hospitals, while primary health care clinics generally had poorer availability (range: 18-48%). Retail or private pharmacies tended to have more survey medicines available (range: 38-62%). There was considerable variability in prices, which tended to be higher in retail pharmacies. The availability of key essential medicines for children was poor. Better understanding of the supply systems in the countries studied and of the pattern of demand for medicines is needed before improvements can be made. Medicines must be available, affordable and acceptable to patients. Substantial progress towards Millennium Development Goals will not occur without a major effort to improve access to medicines for children.


The Bath Adolescent Pain Questionnaire (BAPQ): development and preliminary psychometric evaluation of an instrument to assess the impact of chronic pain on adolescents.
Chronic pain causes significant problems in the lives of many adolescents, considerably affecting their physical, psychological and social functioning. The assessment of the multidimensional impact of chronic pain is an essential clinical task. This study reports on the development and psychometric evaluation of the Bath Adolescent Pain Questionnaire (BAPQ); an assessment tool designed specifically for use with adolescents who experience chronic pain. A sample of 222 adolescents (11-18 years) experiencing chronic pain completed the 109-item draft inventory. Participants were recruited from two different UK clinics. All participants responded to items using a 5-point frequency scale. Psychometric evaluation of the data resulted in a reduced inventory length of 61 items. Internal consistency of all seven questionnaire subscales was established using Cronbach's alpha. Comparative validity was undertaken by comparison of all individual subscales with existing validated measures (SCAS, CDI-S, FDI, Brief FAM. PCS and CASAFS). The temporal reliability of each inventory subscale was established using a sub-sample of 30 adolescent participants over a 17-day period. Psychometric evaluation of the data suggests the inventory yields both a reliable and valid assessment of the impact of chronic pain on the lives of adolescents. The BAPQ may offer a comprehensive way to assess the widespread deleterious impact of adolescent chronic pain in both a research and clinical setting. Further investigation is needed on the predictive validity of the subscales. Additional data from samples of patients with diagnoses that are not musculoskeletal in origin would be of great assistance.


Occupational exposure to zeranol, an animal growth promoter.
Zeranol (3,4,5,6,7,8,9,10,11,12-Decahydro-7,14,16-trihydroxy-3-methyl-1H-2 - benzoxacyclotetradecin-1-one) is a synthetic oestrogenic agent used as an animal growth promoter. The effects of occupational exposure to zeranol in 11 exposed workers from a pelletising plant and 14 nonexposed subjects were assessed. A questionnaire showed that more breast symptoms were reported by male and female plant workers compared with non-exposed subjects, although the difference was not statistically significant. Clinical assessment showed no cases of gynaecomastia in all the male participants. Blood samples analysed by high performance liquid chromatography for zeranol, its precursor zearalenone, and its main metabolites did not show any of these compounds above the laboratory limit of detection. Serum levels of follicle stimulating hormone (FSH), luteinising hormone (LH), prolactin, and oestradiol showed no striking differences between the exposed and the non-exposed subjects. Total and low density lipoprotein cholesterol (LDL cholesterol) levels did not significantly differ between the two groups but mean high density lipoprotein cholesterol (HDL cholesterol) levels were higher in the exposed group; this could be due to relatively high HDL cholesterol in two women exposed to zeranol or relatively low HDL cholesterol in three non-exposed men.


Resistance to fracture of dental roots obturated with different materials.
The aim of this study was to compare the vertical fracture resistance of roots obturated with different root canal filling materials and sealers. Crowns of 55 extracted mandibular premolar teeth were removed to provide root lengths of 13 mm. Five roots were saved as negative control group (canals unprepared and unfilled). Fifty root canals were instrumented and then five roots were saved as positive control group (canals prepared but unfilled). The remaining 45 roots were randomly divided into three experimental groups (n = 15 root/group) and obturated with the following procedures: in group 1, glass ionomer-based sealer and cone (ActiV GP obturation system); in group 2, bioceramic sealer and cone (EndoSequence BC obturation system); and in group 3, roots were filled with bioceramic sealer and cone (Smartpaste bio obturation system). All specimens were tested in a universal testing machine for measuring fracture resistance. For each root, the force at the time of fracture was recorded in Newtons. The statistical analysis was performed by using Kruskal-Wallis and post hoc test. There were no significant differences between the three experimental groups. The fracture values of three experimental and negative control groups were significantly higher than the positive control group. Within the limitations of this study, all materials increased the fracture resistance of instrumented roots.


Personalised risk communication for informed decision making about entering screening programs.
There is a trend towards greater patient involvement in health care decisions. Adequate discussion of the risks and benefits associated with different choices is often required if involvement is to be genuine and effective. Achieving adequate involvement of consumers and informed decision making are now seen as important goals for any screening programme. Individualised risk estimates have been shown to be effective methods of risk communication in general, but the effectiveness of different strategies has not previously been examined. To assess the effects of different types of individualised risk communication for consumers making decisions about participating in screening. We searched the Cochrane Consumers and Communication Review Group specialised register (searched March 2001), MEDLINE (1985 to 2001), EMBASE (1985 to 2001), CancerLit (1985 to 2001), CINAHL (1985 to 2001), ClinPSYC (1989 to 2001), and the Science Citation Index Expanded (searched March 2002). Follow-up searches involved hand searching Preventive Medicine, citation searches on seven authors, and searching reference lists of articles. Randomised controlled trials addressing the decision by consumers of whether or not to undergo screening, incorporating an intervention with a 'personalised risk communication element' and reporting cognitive, affective, or behavioural outcomes. A 'personalised risk communication element' is based on the individual's own risk factors for a condition (such as age or family history). It may be calculated from an individual's risk factors using formulae derived from epidemiological data, and presented as an absolute risk or as a risk score, or it may be categorised into, for example, high, medium or low risk groups. It may be less detailed still, involving a listing, for example, of a consumer's risk factors as a focus for discussion and intervention. Two reviewers independently assessed trial quality and extracted data. Data about the nature and setting of the intervention, and the relevant outcome data were extracted, along with items relating to methodological quality. Thirteen studies were included. Personalised risk communication (whether written, spoken or visually presented) was associated with increased uptake of screening tests (odds ratio (OR) 1.5 (95% confidence interval (CI) 1.11 to 2.03). There was no evidence from these studies that this increase in uptake of tests was related to informed decision making by consumers. More detailed personalised risk communication was associated with a smaller increase in uptake of tests. That is, for personalised risk communication which used and presented numerical calculations of risk, the OR for test uptake was 1.22 (95% CI 0.56 to 2.68). For risk estimates or calculations which were categorised into high, medium or low strata of risk, the OR was 1.42 (95% CI 1.07 to 1.88). For risk communication that simply listed risk personal risk factors the OR was 1.7 (95% CI 1.17 to 2.48). Most of the included studies addressed mammography programmes. These studies showed slightly smaller effects than the overall dataset, again with numerical calculated risk estimates being associated with lower ORs for uptake of tests (OR 1.13; 95% CI 0.98 to 1.29) than the other categories of (less detailed) personalised risk communication. The four studies examining risk communication in high risk individuals showed larger odds ratios for uptake of tests than the other studies. The OR for numerical calculated risk estimates was 1.48 (95% CI 1.06 to 2.07), compared to 4.66 (95% CI 2.24 to 9.69) for categorised risk estimates and 2.64 (95% CI 1.42 to 4.9) for listed personal risk factors. There were insufficient data from the included studies to report odds ratios on other key outcomes such as: intention to take tests, anxiety, satisfaction with decisions, decisional conflict, knowledge and risk perception. Personalised risk communication (as currently implemented in the included studies) is associated with increased uptake of screening programmes, but this may not be interpretable as evidence of informed decision making by consumers.


Association between religiosity and depression varies with age and sex among adults in South America: Evidence from the CESCAS I study.
Prior studies have suggest that religiosity mitigates symptoms of depression. However, population-based data in South America are limited. This study determines the prevalence of religiosity and explores its association with depression in four cities of the Southern cone of Latin-America. In the CESCAS I study 7524 participants aged between 35 and 74 years old were recruited between 2011 and 2012 from randomly selected samples in 4 cities (Bariloche and Marcos Paz, Argentina; Temuco, Chile; and Pando-Barros Blancos, Uruguay). Religiosity was assessed with a questionnaire from the Hispanic Community Health Study/Study of Latinos. Two dimensions were used: 1) recognition as belonging to a religion; and 2) frequency of participation in religious activities. Depression was measured using the PHQ-9. Prevalence of religiosity was described by sociodemographic characteristics. Association between religiosity and depression was examined through logistic regression models controlling for sex, age and other potential confounders. Weekly religious activities were reported by 32.3% (95% CI: 30.1, 33.6) of participants. Prevalence of major depressive episode (MDE) was 14.6% (95% CI: 13.6, 15.6). After controlling for confounders, older women (≥65 years) who reported religious affiliation had 70% lower likelihood of having MDE (OR: 0.3; 95% CI, 0.1, 0.8). Moreover, in this group, women participating in religious activities more than once per week compared with "never" had 50% lower likelihood of having a MDE (OR: 0.5; 95% CI: 0.3, 0.9). No association between religious activities and depression was found in men. Religiosity is highly prevalent among adults in four cities of South America. Our study found an inverse association between religiosity and depression only in women, stronger in olders. Although longitudinal studies are necessary to determine the true nature of these relationships, religiosity may be a relevant factor that health care providers could take into account when exploring depression in their patients.


Risk assessment as a tool for improving external biosecurity at farm level.
Biosecurity routines at herd level may reduce the probability of introduction of disease into the herd, but some measures may be regarded as expensive and cumbersome for the farmers. Custom-made measures based on individual farm characteristics may aid in improving the actual application of on-farm biosecurity. The aim of the study was to provide a tool for calculating the effects of different biosecurity measures and strategies on the individual farm level. A simple model was developed to assess the risk of disease introduction and the need for biosecurity measures in individual farms. To illustrate the general applicability of the tool, it was applied to theoretical examples of Swedish cattle and pig farms and diseases endemic in those animal species in the EU, in two scenarios with different between-farm contact patterns. The model illustrated that the most important factors affecting the risk, and the effect of biosecurity measures such as quarantine routines and protective clothing, were the frequency of between-farm contacts and prevalence of the disease. The risk of introduction as well as the effect of biosecurity measures differed between farm types and disease transmission routes. Adapting contact patterns to mitigate a specific disease risk was as important as biosecurity measures for some farm types, but the largest effect was seen when combining biosecurity measures with more planned contact patterns. The risk assessment model proved useful for illustrating the risk of introduction of endemic diseases and the mitigating effect of different biosecurity measures on farm level. Model outputs could be used to justify prioritisation of measures or adapting contact patterns. The theoretic exercise of adjusting model inputs and comparing outputs may help veterinary advisors to understand farm-specific risks and motivate farmers to improve biosecurity in their individual farm, as it can be tailored to each farmer's needs and preferences.


Cardiac plasticity influences aerobic performance and thermal tolerance in a tropical, freshwater fish at elevated temperatures.
Fishes faced with novel thermal conditions often modify physiological functioning to compensate for elevated temperatures. This physiological plasticity (thermal acclimation) has been shown to improve metabolic performance and extend thermal limits in many species. Adjustments in cardiorespiratory function are often invoked as mechanisms underlying thermal plasticity because limitations in oxygen supply have been predicted to define thermal optima in fishes; however, few studies have explicitly linked cardiorespiratory plasticity to metabolic compensation. Here, we quantified thermal acclimation capacity in the commercially harvested Nile perch (Lates niloticus) of East Africa, and investigated mechanisms underlying observed changes. We reared juvenile Nile perch for 3 months under two temperature regimes, and then measured a series of metabolic traits (e.g. aerobic scope) and critical thermal maximum (CTmax) upon acute exposure to a range of experimental temperatures. We also measured morphological traits of heart ventricles, gills and brains to identify potential mechanisms for compensation. We found that long-term (3 month) exposure to elevated temperature induced compensation in upper thermal tolerance (CTmax) and metabolic performance (standard and maximum metabolic rate, and aerobic scope), and induced cardiac remodeling in Nile perch. Furthermore, variation in heart morphology influenced variations in metabolic function and thermal tolerance. These results indicate that plastic changes enacted over longer exposures lead to differences in metabolic flexibility when organisms are acutely exposed to temperature variation. Furthermore, we established functional links between cardiac plasticity, metabolic performance and thermal tolerance, providing evidence that plasticity in cardiac capacity may be one mechanism for coping with climate change.


Left ventricular thrombus development during ventricular fibrillation and resolution during resuscitation in a swine model of sudden cardiac arrest.
Intracardiac thrombus is a well-known complication of low-flow cardiac states including acute myocardial infarction and atrial fibrillation. Little is known, however, about the formation of intracardiac (left ventricular [LV]) thrombus during the extreme low-flow state of cardiac arrest. Using a swine model of sudden cardiac arrest, we examined the sonographic development of LV thrombus over time after induction of ventricular fibrillation (VF) and resolution of thrombus with cardiopulmonary resuscitation (CPR). This observational study was IACUC approved. Forty-five Yorkshire swine were sedated, intubated, and instrumented under general anesthesia before VF was electrically induced. Sonographic data was collected immediately after VF induction and at 2-min intervals thereafter. Following 12min of untreated VF, resuscitation was initiated with closed chest compressions using an oxygen-powered mechanical resuscitation device. Observations were continued during attempted resuscitation. At the end of the experiment, the animals were euthanized while still at a surgical depth of anesthesia. The data was analyzed descriptively. Sonographic evidence of LV thrombus was observed in 43/45 animals (95.6% [95%CI: 85.2%, 98.8%]). Thrombus was detected within 6min in 39/45 (86.7% [95%CI: 73.8%, 93.8%]) animals that developed thrombus. Thrombus resolved within 2min after initiation of chest compressions in 31/43 (72.1% [95%CI: 57.3%, 83.3%]) animals. Similar to other low-flow cardiac states, LV thrombus develops early in the natural history of VF arrest and resolves quickly once forward flow is re-established by chest compressions. Institutional protocol number: 154600-8.


Prevalence of chronic autoimmune thyroiditis in the urban area neighboring a petrochemical complex and a control area in Sao Paulo, Brazil.
To evaluate the prevalence of chronic autoimmune thyroiditis in 2 urban areas of metropolitan São Paulo (Brazil): Polo Area neighboring a large petrochemical complex and São Bernardo Campo Area (control area). Subjects were randomly included from the adult population (20 to 70 years of age) of both genders (women 80%, men 20%) who voluntarily agreed to participate. From the Polo Area, in the vicinity of a large petrochemical industrial complex, 409 subjects were included; from the control area (São Bernardo Campo Area) 420 individuals were included. All subjects were clinically examined, and a detailed record of past thyroid illness and medications was obtained. Ultrasonographic studies were performed using a portable GE Medical Systems apparatus. Blood samples were obtained for free T4, serum TSH, and serum anti-thyroid peroxidase autoantibodies. Urine specimens were collected in Monovette syringes for assaying iodine content. Salt samples were collected at households, and the iodine content was measured. Chronic autoimmune thyroiditis was diagnosed both echographically (marked hypoechogenicity) and immunologically (presence of autoantibodies against thyroid peroxidase). In the Polo Area, 15.6% of the examined population had chronic autoimmune thyroiditis, and in the control area (São Bernardo Campo Area), 19.5% of the population had evidence of chronic autoimmune thyroiditis (P > 0.057, not significant). The prevalence of hypothyroidism was 4.9% in the Polo Area and 8.3% in the São Bernardo Campo Area (P = 0.0461 significant). Taking the 2 populations together, 6.6% had hypothyroidism (about one third of these patients were on L-T4 treatment). The mean thyroid volume was 11.2 mL. Domestic salt had a normal concentration of iodine (35.5 + 6.61 mg/kg). Urinary excretion of iodine was above 300 microg Iodine/L in 58.5% of the total population. The high iodine intake (above 300 microg Iodine/L of urine) that was present from 1998 through 2005 may be related to a higher prevalence of chronic autoimmune thyroiditis in both areas that were studied. There was no apparent or documented relationship of chronic autoimmune thyroiditis prevalence to the proximity to the petrochemical complex.


Gas-Phase Oxidation of the Protonated Uracil-5-yl Radical Cation.
This study targets the kinetics and product detection of the gas-phase oxidation reaction of the protonated 5-dehydrouracil (uracil-5-yl) distonic radical cation using ion-trap mass spectrometry. Protonated 5-dehydrouracil radical ions (5-dehydrouracilH+ radical ion, m/z 112) are produced within an ion trap by laser photolysis of protonated 5-iodouracil. Storage of the 5-dehydrouracilH+ radical ion in the presence of controlled concentration of O2 reveals two main products. The major reaction product pathway is assigned as the formation of protonated 2-hydroxypyrimidine-4,5-dione (m/z 127) + •OH. A second product ion (m/z 99), putatively assigned as a five-member-ring ketone structure, is tentatively explained as arising from the decarbonylation (-CO) of protonated 2-hydroxypyrimidine-4,5-dione. Because protonation of the 5-dehydrouracil radical likely forms a dienol structure, the O2 reaction at the 5 position is ortho to an -OH group. Following this addition of O2, the peroxyl-radical intermediate isomerizes by H atom transfer from the -OH group. The ensuing hydroperoxide then decomposes to eliminate •OH radical. It is shown that this elimination of •OH radical (-17 Da) is evidence for the presence of an -OH group ortho to the initial phenyl radical site, in good accord with calculations. The subsequent CO loss mechanism, to form the aforementioned five-member-ring structure, is unclear, but some pathways are discussed. By following the kinetics of the reaction, the room temperature second-order rate coefficient of the 5-dehydrouracilH+ distonic radical cation with molecular oxygen is measured at 7.2 × 10-11 cm3 molecule-1 s-1, Φ = 12% (with ±50% total accuracy). For aryl radical reactions with O2, the presence of the •OH elimination product pathway, following the peroxyl-radical formation, is an indicator of an -OH group ortho to the radical site.


RECORD: Reference-Assisted Genome Assembly for Closely Related Genomes.
Background. Next-generation sequencing technologies are now producing multiple times the genome size in total reads from a single experiment. This is enough information to reconstruct at least some of the differences between the individual genome studied in the experiment and the reference genome of the species. However, in most typical protocols, this information is disregarded and the reference genome is used. Results. We provide a new approach that allows researchers to reconstruct genomes very closely related to the reference genome (e.g., mutants of the same species) directly from the reads used in the experiment. Our approach applies de novo assembly software to experimental reads and so-called pseudoreads and uses the resulting contigs to generate a modified reference sequence. In this way, it can very quickly, and at no additional sequencing cost, generate new, modified reference sequence that is closer to the actual sequenced genome and has a full coverage. In this paper, we describe our approach and test its implementation called RECORD. We evaluate RECORD on both simulated and real data. We made our software publicly available on sourceforge. Conclusion. Our tests show that on closely related sequences RECORD outperforms more general assisted-assembly software.


Antithrombotic therapy for atrial fibrillation: Antithrombotic Therapy and Prevention of Thrombosis, 9th ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines.
The risk of stroke varies considerably across different groups of patients with atrial fibrillation (AF). Antithrombotic prophylaxis for stroke is associated with an increased risk of bleeding. We provide recommendations for antithrombotic treatment based on net clinical benefit for patients with AF at varying levels of stroke risk and in a number of common clinical scenarios. We used the methods described in the Methodology for the Development of Antithrombotic Therapy and Prevention of Thrombosis Guidelines: Antithrombotic Therapy and Prevention of Thrombosis, 9th ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines article of this supplement. For patients with nonrheumatic AF, including those with paroxysmal AF, who are (1) at low risk of stroke (eg, CHADS(2) [congestive heart failure, hypertension, age ≥ 75 years, diabetes mellitus, prior stroke or transient ischemic attack] score of 0), we suggest no therapy rather than antithrombotic therapy, and for patients choosing antithrombotic therapy, we suggest aspirin rather than oral anticoagulation or combination therapy with aspirin and clopidogrel; (2) at intermediate risk of stroke (eg, CHADS(2) score of 1), we recommend oral anticoagulation rather than no therapy, and we suggest oral anticoagulation rather than aspirin or combination therapy with aspirin and clopidogrel; and (3) at high risk of stroke (eg, CHADS(2) score of ≥ 2), we recommend oral anticoagulation rather than no therapy, aspirin, or combination therapy with aspirin and clopidogrel. Where we recommend or suggest in favor of oral anticoagulation, we suggest dabigatran 150 mg bid rather than adjusted-dose vitamin K antagonist therapy. Oral anticoagulation is the optimal choice of antithrombotic therapy for patients with AF at high risk of stroke (CHADS(2) score of ≥ 2). At lower levels of stroke risk, antithrombotic treatment decisions will require a more individualized approach.


Upregulation of integrin β4 promotes epithelial-mesenchymal transition and is a novel prognostic marker in pancreatic ductal adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDA) is a highly aggressive and often lethal malignant tumor. Several studies have shown that epithelial-mesenchymal transition (EMT) is frequently observed in clinical samples of PDA and is related to high metastatic rates and poor outcomes. To identify candidate molecules regulating EMT in PDA, we previously used cDNA microarray analysis and identified integrin β4 (ITGB4) as one of the genes upregulated in high-EMT xenografts derived from PDA patients. The aim of the current study was to clarify the clinicopathological and functional significance of ITGB4 overexpression in PDA. ITGB4 upregulation in high-EMT xenografts was confirmed by immunohistochemistry. Immunohistochemical analyses of 134 surgically resected PDA cases revealed intratumoral heterogeneity with respect to ITGB4 expression and showed that cancer cells undergoing EMT often display strong diffuse ITGB4 expression. High levels of ITGB4 expression were significantly correlated with the hallmarks of EMT (solitary cell infiltration, reduced E-cadherin expression, and increased vimentin expression), with high tumor grade, and with the presence of lymph node metastasis, and showed an independent prognostic effect. Immunocytochemical analyses of PDA cell lines revealed that localization of ITGB4 changed from regions of cell-cell contact to diffuse cytoplasm and cell edges with occasional localization in filopodia during EMT. Knockdown of ITGB4 reduced the migratory and invasive ability of PDA cells. Overexpression of ITGB4 promoted cell scattering and cell motility in combination with downregulation of E-cadherin and upregulation of vimentin expression. In conclusion, we elucidated the prognostic and clinicopathological significance of ITGB4 overexpression in PDA and also the potential role for ITGB4 in the regulation of cancer invasion and EMT.


Comparative pharmacology of cisatracurium (51W89), atracurium, and five isomers in cats.
Atracurium has four chiral centers and the marketed product is a mixture of ten optical and geometric isomers. Six of the isomers were prepared and evaluated for neuromuscular blocking activity and autonomic effects in anesthetized cats. This study reports the comparative pharmacology of the six isomers and atracurium that led to the selection of one isomer, cisatracurium (Nimbex, 51W89) for clinical development. Purpose bred cats, anesthetized with alpha-chloralose (80 mg/kg) and pentobarbital sodium (7 mg/kg) administered intraperitoneally, were used in this study. Neuromuscular blocking effects were assessed from the effects on the tibialis anterior twitch evoked at 0.15 Hz. Inhibition of the autonomic nervous system was assessed from the effects on the nictitating membrane contraction, in response to preganglionic sympathetic nerve stimulation and the bradycardia/vasodepressor responses to vagal nerve stimulation. Cardiovascular effects and plasma histamine concentrations were determined after a bolus injection of cisatracurium or atracurium. Like atracurium, all six isomers produced dose-dependent neuromuscular block (NMB). The calculated ED95NMB values varied approximately tenfold (43 +/- 2 microgram/kg -488 +/- 56 microgram/kg. The "R-series" isomers were more potent than the corresponding "S-series" isomers. With the exception of the S,Trans-S', Trans isomers, the NMB effects, i.e., onset times (range 2.6 +/- 0.2 min to 4.7 +/- 0.3 min) and total durations (range 9.9 +/- 1.4 min to 14 +/- 0.9 min), of the other five isomers were very similar to that atracurium. The former isomers had relatively short duration of action. The 25-75% recovery times after cisatracurium at 1 x ED95 (4.4 +/- 0.4 min), 4 x ED95 (4.5 +/- 0.4 min), and continuous infusions lasting at least 60 min that maintained 95-99% NMB (4.8% +/- 0.4 min) indicated a noncumulative effect. The vagal ID50:NMB ED95 ratios for atracurium and the six isomers ranged from 2 to 27. The sympathetic ID25:NMB ED95 ranged from 2.7 to 60. Atracurium and all of the isomers, except cisatracurium, produced cardiovascular effects after intravenous bolus administration at large doses (700-4,800 micrograms/kg). In contrast to atracurium, there were no changes in plasma histamine concentrations associated with the administration of doses of cisatracurium equivalent to 60X the NMB ED95 (62 +/- 8 micrograms/kg). Cisatracurium has neuromuscular blocking effects identical to those of atracurium, is more potent, and does not produce cardiovascular effects or increase plasma histamine concentrations.


Intracellular degradation of chemically functionalized carbon nanotubes using a long-term primary microglial culture model.
Chemically functionalized carbon nanotubes (f-CNTs) have been used in proof-of-concept studies to alleviate debilitating neurological conditions. Previous in vivo observations in brain tissue have suggested that microglia - acting as resident macrophages of the brain - play a critical role in the internalization of f-CNTs and their partial in situ biodegradation following a stereotactic administration in the cortex. At the same time, several reports have indicated that immune cells such as neutrophils, eosinophils and even macrophages could participate in the processing of carbon nanomaterials via oxidation processes leading to degradation, with surface properties acting as modulators of CNT biodegradability. In this study we questioned whether degradability of f-CNTs within microglia could be modulated depending on the type of surface functionalization used. We investigated the kinetics of degradation of multi-walled carbon nanotubes (MWNTs) functionalized via different chemical strategies that were internalized within isolated primary microglia over three months. A cellular model of rat primary microglia that can be maintained in cell culture for a long period of time was first developed. The Raman structural signature of the internalized f-CNTs was then studied directly in cells over a period of up to three months, following a single exposure to a non-cytotoxic concentration of three different f-CNTs (carboxylated, aminated and both carboxylated and aminated). Structural modifications suggesting partial but continuous degradation were observed for all nanotubes irrespective of their surface functionalization. Carboxylation was shown to promote more pronounced structural changes inside microglia over the first two weeks of the study.


Effects of severe global ischemia on N-acetylaspartate and other metabolites in the rat brain.
N-acetylaspartate (NAA) is found exclusively in neurons and their processes in the adult brain. Since the regional distribution of NAA may be imaged using magnetic resonance spectroscopic imaging (1H-MRSI), a regional measure of neuronal density may be noninvasively obtained. The technique may be particularly useful in the diagnosis of diseases where neurons are selectively injured, since these diseases do not result in definitive changes on conventional imaging studies. The goal of this study was to determine whether 1H-MRSI measurement of NAA defects neuronal loss following global ischemia. 1H-MRSI was performed in rats 24 h after global ischemia was induced by bilateral carotid occlusion plus hypotension. 1-H-MRSI showed that NAA was decreased by 28-74% in vulnerable regions, including the cortex, striatum, hippocampus, and, to a lesser extent, the thalamus. No change was observed in the brain stem or cerebellum. Regions where 1H-MRSI observed NAA was decreased also had histological evidence of selective neuronal necrosis and showed marked increase of lactate and alanine. These results show that 1H-MRSI detected loss of NAA in brain regions with selective neuronal loss, suggesting that 1H-MRSI measurements of NAA could detect neuronal loss in a variety of disease states where there is selective neuronal necrosis.


A Mathematical Model of Average Dynamics in a Stem Cell Hierarchy Suggests the Combinatorial Targeting of Cancer Stem Cells and Progenitor Cells as a Potential Strategy against Tumor Growth.
The cancer stem cell hypothesis states that tumors are maintained by a small subpopulation of stem-like cells, often called cancer stem cells (CSCs) or tumor initiating cells. CSCs can self-renew and give rise to more differentiated cells, which comprise the bulk of the tumor. In addition, CSCs are resistant to conventional therapy, which suggests that they are responsible for tumor relapse. This has led researchers to increase efforts to develop directed therapies against CSCs. However, some experiments in mice have shown that the elimination of CSCs might not ensure tumor eradication. This may be due to different events, such as residual CSCs after treatment, the plasticity of cells within the tumor, the presence of different CSCs having their own hierarchy within the same tumor, and the ability of more differentiated cells to maintain the disease, among others. Trying to decipher this complexity may benefit from dissecting the whole in its parts. Here, we hypothesize that tumor relapse after the selective targeting of CSCs may be due to intermediate progenitor (P) cells that can maintain the tumor volume. In order to support the hypothesis, we implemented a mathematical model derived using pseudo-reactions representing the events of each cell subpopulation within the tumor. We aimed to test if a minimal unidirectional hierarchical model consisting of CSCs, P, and terminally differentiated (D) cells could be adjusted to experimental data for selective CSC targeting. We further evaluated therapies ranging from nonselective to specifically directed and combination therapy. We found that selective killing of the CSC compartment has a delaying effect on the overall exponential tumor growth, but was not able to eliminate the disease. We show that therapy that targets both CSCs and intermediate progenitor (P) cells with a sufficient capacity to proliferate and differentiate could represent a more efficient treatment option for tumor depletion. Testing this hypothesis in vivo may allow us to discriminate within the array of possibilities of tumor relapse, and further open the idea of combination therapy against different subpopulations of tumor cells instead of segregating CSCs and bulk tumor cells.


Orthogonal frequency division multiplexing for high-speed optical transmission.
Optical Orthogonal frequency division multiplexing (OOFDM) is shown to outperform RZ-OOK transmission in high-speed optical communications systems in terms of transmission distance and spectral efficiency. The OOFDM in combination with the subcarrier multiplexing offers a significant improvement in spectral efficiency of at least 2.9 bits/s/Hz.