Novel influenza A (H1N1) infection vs. common influenza-like illness: a prospective study.
On June 11th, 2009 the World Health Organization (WHO) declared the first influenza pandemic of the 21st century. Data regarding the clinical characteristics and course of this viral infectious disease are still being assessed. The aim of this study was to investigate and compare the possible differences in clinical course and outcome between H1N1-positive [H1N1(+)] and negative [H1N1(-)] patients. This prospective study was conducted between July 2009 and January 2010 in a regional hospital in Greece. The study population consisted of 165 patients aged 14 years or older, with influenza-like illness (ILI) who, according to CDC recommendations, fulfilled the criteria for diagnostic influenza testing. Enrolled patients underwent a detailed diagnostic work-up. Infection by the H1N1 virus was diagnosed using real-time reverse transcriptase polymerase chain reaction, from pharyngeal swab specimens. We identified 81 H1N1 (+) (49%) patients. Statistical analysis revealed that H1N1(+) patients were significantly younger (median age 27 vs. 35 years, p<0.05), had a decreased white blood cell count (median 7.200 vs. 8.415, p<0.05) and an increased percentage of monocytes (55.6% vs. 27.4%, p<0.05) compared to the H1N1(-) patients. The clinical presentation at the emergency department, as well as the hospital admission and disease complication rate, were not significantly different between the 2 groups. The clinical characteristics of the new influenza virus appear to be mild and to resemble those of common influenza-like illnesses (ILI). The patients who tested positive for the H1N1 virus were younger and had an increased percentage of monocytes compared to the H1N1-negative patients.


Effects of oral intake of heat-killed Lactobacillus brevis SBC8803 (SBL88™) on dry skin conditions: A randomized, double-blind, placebo-controlled study.
Lactobacilli are important in intestinal homeostasis, which involves the regulation of immune function, digestive health, cholesterol absorption and intestinal tumor growth amongst others. Our previous investigations have suggested that oral intake of heat-killed Lactobacillus brevis (L. brevis) SBC8803 (SBL88™) suppresses dermatitis by modulating the immune function in an atopic dermatitis mouse model. The aim of the present study was to investigate the effect of heat-killed L. brevis SBC8803 intake on skin hydration conditions in humans. A randomized, double-blind, placebo-controlled study was conducted with volunteers with slightly higher levels of transepidermal water loss (TEWL) on the forearm. The subjects (126 people aged between 21 and 59 years) were randomly allocated to three groups so that the level of TEWL and the age were distributed equally among the groups. The subjects took placebo or heat-killed L. brevis SBC8803 at a daily dose of 25 or 50 mg for 12 weeks. Following the exclusion of eight subjects for plausible reasons (two withdrawals from the study, two for study violations, one for not meeting exclusion criteria and three due to their physical condition), 118 subjects were subjected to the analysis. The results of the present study revealed that following the analysis of the whole populations, marginal differences were observed in TEWL (for example, suppression of skin water loss) at the neck in the 25 mg/day group at week 8 and at the lower eye region in the 50 mg/day group at week 4 (P=0.05 and 0.09, respectively, compared with the placebo group analyzed by Dunnett's test). A significant increase in corneal hydration was also observed at the neck in the 25 mg/day group at week 12 (P=0.06, as compared with the placebo group as analyzed by Dunnett's test). In the analysis of the subpopulations whose habitual frequency of taking lactic fermentation products was less than once per week, the levels of corneal hydration at the neck (in the 50 mg/day group) and lower eye region (in the 25 mg/day group) were significantly increased at week 12 (P<0.05). In conclusion, the results of the present investigation suggest that oral intake of heat-killed L. brevis SBC8803 is effective at improving skin hydration conditions in populations with low habitual frequency of taking lactic fermentation products.


Analysis of learning deficits in aged rats on the W-track continuous spatial alternation task.
Young and aged animals were tested on a spatial alternation task that consisted of two interleaved components: (1) an "outbound" or alternation component (working memory) and (2) an "inbound" component, requiring the animal to remember to return to a central location in space (spatial memory). In the present study, aged rats made more outbound errors throughout testing, resulting in significantly more days to reach learning criterion, as compared to young rats. Furthermore, while all animals were able to learn the hippocampus-dependent inbound component of the task, most aged animals remained just above chance on the outbound component, even after extended testing days. Aged rats may be more impaired on the outbound part of the task because it requires cooperation of both the hippocampus and mPFC, each of which is compromised with age. In addition to presenting these results, we compare one commonly used analysis (repeated measures ANOVA) and two less common hierarchical modeling techniques (hierarchical generalized linear model and state-space random effects model) to determine the best method for comparing population learning over time. We found that hierarchical modeling is the most appropriate for this task and that a state-space model better captures the behavioral responses. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Free amino acid pool and muscle protein balance after resistance exercise.
The aim of this study was to assess the effects of a resistance exercise session (RES) on free amino acid concentrations and protein synthesis and breakdown of the vastus lateralis (VL) muscle during recovery in male subjects. Both the exercise group (EG) and the control group (CG) consisted of six healthy physically active men. On the experiment day in fasting conditions, a stable isotopic tracer of L-[ring-2H(5)] phenylalanine was infused and EG started a heavy 50-min hypertrophic RES for lower extremities after 55 min of infusion. At the same time, CG was at rest. During recovery of 195 min after RES, several blood samples were drawn from the femoral artery (FA) and the femoral vein (FV) and muscle samples from the VL muscle. The enrichment was analyzed by GC/MS and leg muscle amino acid kinetics determined by three-pool compartment model between FA, FV, and VL. During recovery at 60 min after RES, there was no difference in muscle protein synthesis or muscle protein breakdown between the groups, but at 195 min, both muscle protein synthesis (P < 0.05) and muscle protein breakdown (P < 0.05) were increased in EG compared with CG. The protein net balance was negative and similar in both groups. Simultaneously in serum concentrations, there was a decrease in leucine (P < 0.05) associated with an increase in aspartate (P < 0.05). Furthermore, the exercise-induced increase in alanine concentration decreased both in serum and muscle. In fasting conditions, protein net balance is negative and RES induces an increase in muscle protein synthesis and breakdown at 195 min but not yet at 60 min of recovery.


Ligand binding and protein dynamics in lactate dehydrogenase.
Recent experimental studies suggest that lactate dehydrogenase (LDH) binds its substrate via the formation of a LDH/NADH.substrate encounter complex through a select-fit mechanism, whereby only a minority population of LDH/NADH is binding-competent. In this study, we perform molecular dynamics calculations to explore the variations in structure accessible to the binary complex with a focus on identifying structures that seem likely to be binding-competent and which are in accord with the known experimental characterization of forming binding-competent species. We find that LDH/NADH samples quite a range of protein conformations within our 2.148 ns calculations, some of which yield quite facile access of solvent to the active site. The results suggest that the mobile loop of LDH is perhaps just partially open in these conformations and that multiple open conformations, yielding multiple binding pathways, are likely. These open conformations do not require large-scale unfolding/melting of the binary complex. Rather, open versus closed conformations are due to subtle protein and water rearrangements. Nevertheless, the large heat capacity change observed between binding-competent and binding-incompetent can be explained by changes in solvation and an internal rearrangement of hydrogen bonds. We speculate that such a strategy for binding may be necessary to get a ligand efficiently to a binding pocket that is located fairly deep within the protein's interior.


Congenital secretory diarrhoea caused by activating germline mutations in GUCY2C.
Congenital sodium diarrhoea (CSD) refers to a form of secretory diarrhoea with intrauterine onset and high faecal losses of sodium without congenital malformations. The molecular basis for CSD remains unknown. We clinically characterised a cohort of infants with CSD and set out to identify disease-causing mutations by genome-wide genetic testing. We performed whole-exome sequencing and chromosomal microarray analyses in 4 unrelated patients, followed by confirmatory Sanger sequencing of the likely disease-causing mutations in patients and in their family members, followed by functional studies. We identified novel de novo missense mutations in GUCY2C, the gene encoding receptor guanylate cyclase C (GC-C) in 4 patients with CSD. One patient developed severe, early-onset IBD and chronic arthritis at 4 years of age. GC-C is an intestinal brush border membrane-bound guanylate cyclase, which functions as receptor for guanylin, uroguanylin and Escherichia coli heat-stable enterotoxin. Mutations in GUCY2C were present in different intracellular domains of GC-C, and were activating mutations that enhanced intracellular cyclic guanosine monophosphate accumulation in a ligand-independent and ligand-stimulated manner, following heterologous expression in HEK293T cells. Dominant gain-of-function GUCY2C mutations lead to elevated intracellular cyclic guanosine monophosphate levels and could explain the chronic diarrhoea as a result of decreased intestinal sodium and water absorption and increased chloride secretion. Thus, mutations in GUCY2C indicate a role for this receptor in the pathogenesis of sporadic CSD.


Recirculatory spinal subarachnoid perfusions in dogs: a method for determining CSF dynamics under non-steady state conditions.
Open-ended ventriculocisternal perfusion techniques for determining cerebrospinal fluid production and absorption rates are severely restricted by the absolute requirement that steady state conditions be present. A new closed recirculatory spinal perfusion technique is described. Because steady state equilibrium is not necessary, numerous determinations at multiple intracranial pressures or under varied experimental conditions are possible within relatively brief perfusion periods. Cerebrospinal fluid (CSF) and nondiffusible albumin tracer are rapidly recirculated through the spinal subarachnoid space in a cephalad direction. The concentration of fluorescein-tagged albumin is continuously monitored as the CSF is circulated through a fluorometric flow cell. Measured continuously, intracranial pressure (ICP) is regulated by changing the volume of the external perfusion circuit with a syringe pump connected in series to the recirculatory spinal perfusion. CSF formation and absorption rates are calculated from measurements of albumin concentration, concentration changes with time, ICP, syringe pump infusion rate, and the external perfusion circuit volume. In dogs, data can be collected after only 45 minutes for mixing; perfusions at four or five intracranial pressures in addition to normal resting pressure can be completed within 2 to 3 hours. The data from 15 perfusions in 14 dogs are presented. The method provides normal resting pressure values of CSF production and absorption consistent with those values in the literature determined by traditional ventriculocisternal perfusion techniques. Determinations at multiple intracranial pressures suggest a proportional relationship between absorption and ICP. No consistent acute change in CSF formation with pressures to 50 mm Hg can be inferred from these data.


A Systematic Review on the Pharmacokinetics of Cannabidiol in Humans.
Background: Cannabidiol is being pursued as a therapeutic treatment for multiple conditions, usually by oral delivery. Animal studies suggest oral bioavailability is low, but literature in humans is not sufficient. The aim of this review was to collate published data in this area. Methods: A systematic search of PubMed and EMBASE (including MEDLINE) was conducted to retrieve all articles reporting pharmacokinetic data of CBD in humans. Results: Of 792 articles retireved, 24 included pharmacokinetic parameters in humans. The half-life of cannabidiol was reported between 1.4 and 10.9 h after oromucosal spray, 2-5 days after chronic oral administration, 24 h after i.v., and 31 h after smoking. Bioavailability following smoking was 31% however no other studies attempted to report the absolute bioavailability of CBD following other routes in humans, despite i.v formulations being available. The area-under-the-curve and Cmax increase in dose-dependent manners and are reached quicker following smoking/inhalation compared to oral/oromucosal routes. Cmax is increased during fed states and in lipid formulations. Tmax is reached between 0 and 4 h. Conclusions: This review highlights the paucity in data and some discrepancy in the pharmacokinetics of cannabidiol, despite its widespread use in humans. Analysis and understanding of properties such as bioavailability and half-life is critical to future therapeutic success, and robust data from a variety of formulations is required.


Quit Experiences among Primary Care Patients Enrolled in a Smoking Cessation Pilot RCT Early in the COVID-19 Pandemic.
The impact of the COVID-19 pandemic on US adults' smoking and quitting behaviors is unclear. We explored the impact of COVID-19 on smoking behaviors, risk perceptions, and reactions to text messages during a statewide stay-at-home advisory among primary care patients who were trying to quit. From May-June 2020, we interviewed smokers enrolled in a 12-week, pilot cessation trial providing text messaging and mailed nicotine replacement medication (NCT04020718). Twenty-two individuals (82% white, mean age 55 years), representing 88% of trial participants during the stay-at-home advisory, completed exit interviews; four (18%) of them reported abstinence. Interviews were thematically analyzed by two coders. COVID-19-induced environmental changes had mixed effects, facilitating quitting for some and impeding quitting for others. While stress increased for many, those who quit found ways to cope with stress. Generally, participants felt at risk for COVID-19 complications but not at increased risk of becoming infected. Reactions to COVID-19 and quitting behaviors differed across age groups, older participants reported difficulties coping with isolation (e.g., feeling disappointed when a text message came from the study and not a live person). Findings suggest that cessation interventions addressing stress and boredom are needed during COVID-19, while smokers experiencing isolation may benefit from live-person supports.


Automated linkage of proteins and payloads producing monodisperse conjugates.
Controlled protein functionalization holds great promise for a wide variety of applications. However, despite intensive research, the stoichiometry of the functionalization reaction remains difficult to control due to the inherent stochasticity of the conjugation process. Classical approaches that exploit peculiar structural features of specific protein substrates, or introduce reactive handles via mutagenesis, are by essence limited in scope or require substantial protein reengineering. We herein present equimolar native chemical tagging (ENACT), which precisely controls the stoichiometry of inherently random conjugation reactions by combining iterative low-conversion chemical modification, process automation, and bioorthogonal trans-tagging. We discuss the broad applicability of this conjugation process to a variety of protein substrates and payloads.


Health Promotion Interventions for Low-Income Californians Through Medi-Cal Managed Care Plans, 2012.
Prevention is the most cost-effective approach to promote population health, yet little is known about the delivery of health promotion interventions in the nation's largest Medicaid program, Medi-Cal. The purpose of this study was to inventory health promotion interventions delivered through Medi-Cal Managed Care Plans; identify attributes of the interventions that plans judged to have the greatest impact on their members; and determine the extent to which the plans refer members to community assistance programs and sponsor health-promoting community activities. The lead health educator from each managed care plan was asked to complete a 190-item online survey in January 2013; 20 of 21 managed care plans responded. Survey data on the health promotion interventions with the greatest impact were grouped according to intervention attributes and measures of effectiveness; quantitative data were analyzed using descriptive statistics. Health promotion interventions judged to have the greatest impact on Medi-Cal members were delivered in various ways; educational materials, one-on-one education, and group classes were delivered most frequently. Behavior change, knowledge gain, and improved disease management were cited most often as measures of effectiveness. Across all interventions, median educational hours were limited (2.4 h), and median Medi-Cal member participation was low (265 members per intervention). Most interventions with greatest impact (120 of 137 [88%]) focused on tertiary prevention. There were mixed results in referring members to community assistance programs and investing in community activities. Managed care plans have many opportunities to more effectively deliver health promotion interventions. Establishing measurable, evidence-based, consensus standards for such programs could facilitate improved delivery of these services.


Ras- and PI3K-dependent breast tumorigenesis in mice and humans requires focal adhesion kinase signaling.
Cancer cells require sustained oncogenic signaling in order to maintain their malignant properties. It is, however, unclear whether they possess other dependencies that can be exploited therapeutically. We report here that in a large fraction of human breast cancers, the gene encoding focal adhesion kinase (FAK), a core component of integrin signaling, was amplified and FAK mRNA was overexpressed. A mammary gland-specific deletion of Fak in mice did not seem to affect normal mammary epithelial cells, and these mice were protected from tumors initiated by the polyoma middle T oncoprotein (PyMT), which activates Ras and PI3K. FAK-deficient PyMT-transformed cells displayed both growth arrest and apoptosis, as well as diminished invasive and metastatic capacity. Upon silencing of Fak, mouse mammary tumor cells transformed by activated Ras became senescent and lost their invasive ability. Further, Neu-transformed cells also underwent growth arrest and apoptosis if integrin beta4-dependent signaling was simultaneously inactivated. Human breast cancer cells carrying oncogenic mutations that activate Ras or PI3K signaling displayed similar responses upon silencing of FAK. Mechanistic studies indicated that FAK sustains tumorigenesis by promoting Src-mediated phosphorylation of p130Cas. These results suggest that FAK supports Ras- and PI3K-dependent mammary tumor initiation, maintenance, and progression to metastasis by orchestrating multiple core cellular functions, including proliferation, survival, and avoidance of senescence.


Association of Somatic Driver Alterations With Prognosis in Postmenopausal, Hormone Receptor-Positive, HER2-Negative Early Breast Cancer: A Secondary Analysis of the BIG 1-98 Randomized Clinical Trial.
A range of somatic driver alterations has been described in estrogen receptor-positive, HER2-negative (ER+/HER2-) early breast cancer (BC); however, the clinical relevance is unknown. To investigate associations of driver alterations with prognosis and the role of PIK3CA mutations in prediction of benefit associated with endocrine therapy in postmenopausal patients with ER+/HER2- early BC treated with tamoxifen or letrozole. The Breast International Group (BIG) 1-98 trial randomized 8010 postmenopausal patients with hormone receptor-positive, operable, invasive BC to monotherapy with letrozole, tamoxifen, or a sequential strategy for 5 years. Driver alterations were characterized using next-generation sequencing in primary tumors from a subset of 764 patients from 7329 eligible patients with ER+/HER2- BC, with 841 distant recurrences after a median of 8.1 years of follow-up. To correct for the oversampling of distant recurrences, weighted analysis methods were used. This analysis was conducted from April 4, 2016, to November 30, 2016. The prevalence of driver alterations, associations with clinicopathologic factors, distant recurrence-free interval, and treatment interactions were analyzed. Multivariable analyses were performed to adjust for clinicopathologic factors. Of 764 samples, 538 (70.4%), including 140 distant recurrence events, were successfully sequenced. Nineteen driver alterations were observed with 5% or greater frequency, with a mean of 4 alterations (range, 0-15) per tumor. PIK3CA mutations were the most common (49%) and were significantly associated with reduction in the risk for distant recurrence (hazard ratio [HR], 0.57; 95% CI, 0.38-0.85; P = .006). TP53 mutations (HR, 1.92; 95% CI, 1.21-3.04; P = .006), amplifications on 11q13 (HR, 2.14; 95% CI, 1.36-3.37; P = .001) and 8p11 (HR, 3.02; 95% CI, 1.88-4.84; P < .001), and increasing number of driver alterations (HR per additional alteration, 1.18; 95% CI, 1.11-1.25; P < .001) were associated with significantly greater risk. Amplifications on 11q13 and 8p11 remained significant predictors in multivariable analysis, but not PIK3CA and TP53 mutations. Patients with tumors harboring kinase or helical domain PIK3CA mutations derived significantly greater benefit from letrozole over tamoxifen than patients whose tumors did not (P interaction = .002). In ER+/HER2- postmenopausal, early-stage BC, amplifications on 11q13 and 8p11 were significantly associated with increased risk for distant recurrence and PIK3CA mutations were predictive of greater magnitude of benefit from letrozole. With these findings, DNA-based classification may aid adjuvant treatment decision making in this setting. ClinicalTrials.gov Identifier: NCT00004205.


The transcription factor Mesp1 interacts with cAMP-responsive element binding protein 1 (Creb1) and coactivates Ets variant 2 (Etv2) gene expression.
Mesoderm posterior 1 (Mesp1) is well recognized for its role in cardiac development, although it is expressed broadly in mesodermal lineages. We have previously demonstrated important roles for Mesp1 and Ets variant 2 (Etv2) during lineage specification, but their relationship has not been defined. This study reveals that Mesp1 binds to the proximal promoter and transactivates Etv2 gene expression via the CRE motif. We also demonstrate the protein-protein interaction between Mesp1 and cAMP-responsive element binding protein 1 (Creb1) in vitro and in vivo. Utilizing transgenesis, lineage tracing, flow cytometry, and immunostaining technologies, we define the lineage relationship between Mesp1- and Etv2-expressing cell populations. We observe that the majority of Etv2-EYFP(+) cells are derived from Mesp1-Cre(+) cells in both the embryo and yolk sac. Furthermore, we observe that the conditional deletion of Etv2, using a Mesp1-Cre transgenic strategy, results in vascular and hematopoietic defects similar to those observed in the global deletion of Etv2 and that it has embryonic lethality by embryonic day 9.5. In summary, our study supports the hypothesis that Mesp1 is a direct upstream transactivator of Etv2 during embryogenesis and that Creb1 is an important cofactor of Mesp1 in the transcriptional regulation of Etv2 gene expression.


Pancreas transplantation: indications, clinical management, and outcomes.
Although many advances have been made, pancreas transplantation still poses several challenges to the surgeon, internist and patient. With success rates now above 80% and improving yearly, diabetic patients must make a major life-style decision when considering a pancreas transplant. The main concerns are will the benefits of insulin-independence off-set the risks of surgery and immunosuppression. For diabetics near dialysis and considering a kidney transplant, the decision may not be as difficult. However, for those patients who are failing insulin therapy (brittle control) and remain with good renal function, the options are limited. As the success of pancreas transplantation improves, the procedure may become routine at more centers and become accepted by more third-party carriers. However, as with other solid organs, the availability of pancreases is limited and the supply soon to be exhausted. Thus, further advances are required for the prevention and treatment of Type 1 diabetes. Hopefully, the new frontiers of the next century will allow physicians to identify and preventively treat those at risk for the development of diabetes. Thus, the population of patients suffering from the consequences of this dreadful disease will be greatly reduced. With new developments in immunosuppression and islet transplantation, diabetic patients of the future may be offered the option of a procedure with reduced risks, less morbidity, and improved long-term cure rates.


Factors affecting psychiatric inpatient hospitalization from a psychiatric emergency service.
As a gateway to the mental health system, psychiatric emergency services (PES) are charged with assessing a heterogeneous array of short-term and long-term psychiatric crises. However, few studies have examined factors associated with inpatient psychiatric hospitalization following PES in a racially diverse sample. We examine the demographic, service use and clinical factors associated with inpatient hospitalization and differences in predisposing factors by race and ethnicity. Three months of consecutive admissions to San Francisco's only 24-h PES (N = 1,305) were reviewed. Logistic regression was used to estimate the associations between demographic, service use, and clinical factors and inpatient psychiatric hospitalization. We then estimated separate models for Asians, Blacks, Latinos and Whites. Clinical severity was a consistent predictor of hospitalization. However, age, gender, race/ethnicity, homelessness and employment status were all significant related to hospitalization. Alcohol and drug use were associated with lower probability of inpatient admission, however specific substances appear particularly salient for different racial/ethnic groups. While clinical characteristics played an essential role in disposition decisions, these results point to the importance of factors external to PES. Individual and community factors that affect use of psychiatric emergency services merit additional focused attention.


Detection of infectious hematopoietic necrosis virus in an invertebrate (Callibaetis sp).
Infectious hematopoietic necrosis virus (IHNV) was detected in the common Mayfly (Callibaetis sp) by western immunoblot assay and was propagated in fish cells (CHSE-214) in culture. When propagated in cell culture, cytopathic effect characteristic of IHNV infection was observed. Antibody specific for IHNV was used to detect all of the major proteins of IHNV in the western immunoblot assay. When the crude lysate was subjected to electron microscopy, bullet-shaped particles (84 nm x 194 nm) characteristic of IHNV were observed. The data suggested that the Mayfly may be a factor in the dissemination of IHNV.


Multimetallic cooperativity in uranium-mediated CO₂ activation.
The metal-mediated redox transformation of CO2 in mild conditions is an area of great current interest. The role of cooperativity between a reduced metal center and a Lewis acid center in small-molecule activation is increasingly recognized, but has not so far been investigated for f-elements. Here we show that the presence of potassium at a U, K site supported by sterically demanding tris(tert-butoxy)siloxide ligands induces a large cooperative effect in the reduction of CO2. Specifically, the ion pair complex [K(18c6)][U(OSi(O(t)Bu)3)4], 1, promotes the selective reductive disproportionation of CO2 to yield CO and the mononuclear uranium(IV) carbonate complex [U(OSi(O(t)Bu)3)4(μ-κ(2):κ(1)-CO3)K2(18c6)], 4. In contrast, the heterobimetallic complex [U(OSi(O(t)Bu)3)4K], 2, promotes the potassium-assisted two-electron reductive cleavage of CO2, yielding CO and the U(V) terminal oxo complex [UO(OSi(O(t)Bu)3)4K], 3, thus providing a remarkable example of two-electron transfer in U(III) chemistry. DFT studies support the presence of a cooperative effect of the two metal centers in the transformation of CO2.


Pain acceptance and its impact on function and symptoms in fibromyalgia.
Objectives Fibromyalgia is a chronic widespread pain (CWP) syndrome of unknown etiology with substantial burden of illness and functional impairment. Pain acceptance has emerged as an interesting target of therapy in chronic pain populations, but few studies have yet been done on the effect of pain acceptance on patients with fibromyalgia. The aim of the present study was to examine the relationship between pain acceptance and its impact on function and symptoms in fibromyalgia with both a cross-sectional and longitudinal design. Methods Three hundred and sixty five participants aged 22-70 with fibromyalgia were recruited from the Norwegian Fibromyalgia Association (NFA). They filled out a questionnaire containing the Fibromyalgia Impact Questionnaire (FIQ), measurement of function and symptoms, and Chronic Pain Acceptance Questionnaire (CPAQ), measurement of pain acceptance, in addition to sociodemographic and clinical variables such as degree of fibromyalgia, depression and pain duration (T1 measures). One year after, 87 of the participants filled out the FIQ and clinical measures once again (T2 measures). Unadjusted and adjusted linear regression analyses were performed both for cross-sectional measures at T1 and for longitudinal measures from T1 to T2, with FIQ score as the outcome variable and CPAQ score at T1 as one of the main independent variables. Results Higher CPAQ score was significantly associated with a lower FIQ score at T1, also when adjusting for age, education, work, depression and Fibromyalgianess Score (p<0.01). Lower FIQ score indicate less impact of fibromyalgia on functioning. In addition, two adjusted linear regression models found higher pain acceptance (CPAQ score) at T1 to be associated with lower negative impact of fibromyalgia on function and symptoms (FIQ score) at T2 (p<0.01). Conclusions Higher pain acceptance is associated with better functional level and less symptoms in fibromyalgia, both cross-sectionally and when measurements are separated in time. Further research should include experimental studies with acceptance-based interventions for this patient group.


Impact of unintentional selective harvesting on the population dynamics of red grouse.
1. The effect of selective exploitation of certain age, stage or sex classes (e.g., trophy hunting) on population dynamics is relatively well studied in fisheries and sexually dimorphic mammals. 2. Harvesting of terrestrial species with no morphological differences visible between the different age and sex classes (monomorphic species) is usually assumed to be nonselective because monomorphicity makes intentionally selective harvesting pointless and impractical. But harvesting of the red grouse (Lagopus lagopus scoticus), a monomorphic species, was recently shown to be unintentionally selective. This study uses a sex- and age-specific model to explore the previously unresearched effects of unintentional harvesting selectivity. 3. We examine the effects of selectivity on red grouse dynamics by considering models with and without selectivity. Our models include territoriality and parasitism, two mechanisms known to be important for grouse dynamics. 4. We show that the unintentional selectivity of harvesting that occurs in red grouse decreases population yield compared with unselective harvesting at high harvest rates. Selectivity also dramatically increases extinction risk at high harvest rates. 5. Selective harvesting strengthens the 3- to 13-year red grouse population cycle, suggesting that the selectivity of harvesting is a previously unappreciated factor contributing to the cycle. 6. The additional extinction risk introduced by harvesting selectivity provides a quantitative justification for typically implemented 20-40% harvest rates, which are below the maximum sustainable yield that could be taken, given the observed population growth rates of red grouse. 7. This study shows the possible broad importance of investigating in future research whether unintentionally selective harvesting occurs on other species.


Bladder management and risk of bladder stone formation in spinal cord injured patients.
We determined by statistical analysis whether method of management is associated with risk of bladder stone formation in spinal cord injured patients. A retrospective cohort study was performed of 457 patients admitted to Stoke Mandeville Hospital Spinal Injuries Center between 1985 and 1990 with more than 6 months of followup. Analysis included Cox regression and Poisson regression. Relative to those patients treated with intermittent self-catheterization, the hazard ratio was 10.5 (p <0.0005, 95% confidence interval 4.0-27.5) for patients with suprapubic catheters and it was 12.8 (p <0.0005, 95% confidence interval 5.1-31.9) for those with indwelling urethral catheters. The absolute annual risk of stone formation in patients with a catheter was 4% compared with 0.2% for those on intermittent self-catheterization. However, having formed a stone, the risk of forming a subsequent stone quadrupled to 16% per year. Bladder stones were no more likely to form in patients with suprapubic catheters compared to those with indwelling urethral catheters (hazard ratio 1.2, p = 0.6). In spinal cord injured patients long-term catheterization is associated with a substantial increased risk of bladder stone formation. This increased risk occurs independently of age, sex and injury level. Degree of injury (complete or incomplete) was considered in the model. Catheter type (suprapubic or urethral) did not change this risk significantly if at all.


Increased expression of the Mr 72,000 type IV collagenase in human colonic adenocarcinoma.
Proteolytic enzymes, such as type IV collagenase, play an important role in tumor invasion and metastasis. To examine Mr 72,000 type IV collagenase expression in human colon carcinoma, blot hybridization of total RNA from 19 primary colon tumors were performed. These filters were probed with complementary DNA probes encoding the Mr 72,000 type IV collagenase metalloenzyme. The results were expressed as the ratio of the messenger RNA (mRNA) levels in the tumor tissue to that in the adjacent normal mucosa (R). The level of the 3.1-kilobase type IV collagenase mRNA was higher in the primary tumor than in the normal adjacent colonic mucosa in 13 of 18 (72%) cases with a diagnosis of adenocarcinoma. These cases were divided into high expression (R, 4.50 to 29.34) and intermediate expression (R, 2.54 to 3.31) subgroups. Both groups showed statistically significant (P less than 0.05) elevations when compared with the five cases showing the lowest levels of Mr 72,000 type IV collagenase mRNA expression (low expression subgroup; R, 0.96 to 1.48). With this demonstrated elevation of Mr 72,000 type IV collagenase mRNA in colorectal adenocarcinoma we examined concomitant expression at the protein level using immunohistochemical techniques. Immunohistochemical examination of 70 cases of colon tumors, including 30 benign adenomas, using anti-Mr 72,000 type IV collagenase antibodies demonstrated a significant correlation with Duke's classification (P less than 0.001). Our results suggest that enhanced expression of the Mr 72,000 type IV collagenase enzyme may be a marker of human colorectal tumor invasiveness.


Keratinocytes negatively regulate the N-cadherin levels of melanoma cells via contact-mediated calcium regulation.
In human skin, melanocytes and their neighboring keratinocytes have a close functional interrelationship. Keratinocytes, which represent the prevalent cell type of human skin, regulate melanocytes through various mechanisms. Here, we use a keratinocyte and melanoma co-culture system to show for the first time that keratinocytes regulate the cell surface expression of N-cadherin through cell-cell contact. Compared to mono-cultured human melanoma A375 cells, which expressed high levels of N-cadherin, those co-cultured with the HaCaT human keratinocyte cell line showed reduced levels of N-cadherin. This reduction was most evident in areas of A375 cells that underwent cell-cell contact with the HaCaT cells, whereas HaCaT cell-derived extracellular matrix and conditioned medium both failed to reduce N-cadherin levels. The intracellular level of calcium in co-cultured A375 cells was lower than that in mono-cultured A375 cells, and treatment with a cell-permeant calcium chelator (BAPTA) reduced the N-cadherin level of mono-cultured A375 cells. Furthermore, co-culture with HaCaT cells reduced the expression levels of transient receptor potential cation channel (TRPC) 1, -3 and -6 in A375 cells, and siRNA-mediated multi-depletion of TRPC1, -3 and -6 reduced the N-cadherin level in these cells. Taken together, these data suggest that keratinocytes negatively regulate the N-cadherin levels of melanoma cells via cell-to-cell contact-mediated calcium regulation.


He-Ne and cw CO2 laser long-path systems for gas detection.
This paper describes the design and testing of a laboratory prototype dual He-Ne laser system for the detection of methane leaks from underground pipelines and solid-waste landfill sites using differential absorption of radiation backscattered from topographic targets. A laboratory-prototype dual cw carbon dioxide laser system also using topographic backscatter is discussed, and measurement results for methanol are given. With both systems, it was observed that the time-varying differential absorption signal was useful in indicating the presence of a gas coming from a nearby source. Limitations to measurement sensitivity, especially the role of speckle and atmospheric turbulence, are described. The speckle results for hard targets are contrasted with those from atmospheric aerosols. The Appendix gives appropriate laser lines and values of absorption coefficients for the hydrazine fuel gases.


LRIG1 opposes epithelial-to-mesenchymal transition and inhibits invasion of basal-like breast cancer cells.
LRIG1 (leucine-rich repeat and immunoglobulin-like domain containing), a member of the LRIG family of transmembrane leucine-rich repeat-containing proteins, is a negative regulator of receptor tyrosine kinase signaling and a tumor suppressor. LRIG1 expression is broadly decreased in human cancer and in breast cancer and low expression of LRIG1 has been linked to decreased relapse-free survival. Recently, low expression of LRIG1 was revealed to be an independent risk factor for breast cancer metastasis and death. These findings suggest that LRIG1 may oppose breast cancer cell motility and invasion, cellular processes that are fundamental to metastasis. However, very little is known of LRIG1 function in this regard. In this study, we demonstrate that LRIG1 is downregulated during epithelial-to-mesenchymal transition (EMT) of human mammary epithelial cells, suggesting that LRIG1 expression may represent a barrier to EMT. Indeed, depletion of endogenous LRIG1 in human mammary epithelial cells expands the stem cell population, augments mammosphere formation and accelerates EMT. Conversely, expression of LRIG1 in highly invasive Basal B breast cancer cells provokes a mesenchymal-to-epithelial transition accompanied by a dramatic suppression of tumorsphere formation and a striking loss of invasive growth in three-dimensional culture. LRIG1 expression perturbs multiple signaling pathways and represses markers and effectors of the mesenchymal state. Furthermore, LRIG1 expression in MDA-MB-231 breast cancer cells significantly slows their growth as tumors, providing the first in vivo evidence that LRIG1 functions as a growth suppressor in breast cancer.


Aldose reductase inhibition increases CNTF-like bioactivity and protein in sciatic nerves from galactose-fed and normal rats.
The impact of exaggerated polyol pathway flux on ciliary neurotrophic factor (CNTF)-like bioactivity and expression of CNTF in rat sciatic nerve was examined after 2 months of galactose intoxication. Polyol content was elevated (P < 0.001) and motor nerve conduction velocity reduced (P < 0.05) in galactose-fed rats compared with control animals or control and galactose-fed rats treated with the aldose reductase inhibitor (ARI) Ponalrestat. CNTF-like bioactivity in the galactose-fed group was reduced to 30% of that assayed in the control group (P < 0.001). ARI treatment significantly increased CNTF-like bioactivity by 60% compared with the untreated galactose group (P < 0.05) but did not restore it to control levels. Unexpectedly, bioactivity in ARI-treated control animals was increased by nearly 250% compared with untreated controls (P < 0.005). In addition to the deficit in CNTF bioactivity in untreated galactose rats, the expression of protein, but not of mRNA, was reduced (P < 0.05). In ARI-treated control and galactose-fed rats, the expression of CNTF peptide was significantly enhanced above control levels (both P < 0.05). Concomitant with the reduction in CNTF levels, there was a shift in the axonal size-frequency distribution of myelinated fibers toward smaller axons in galactose-fed rats that was prevented by ARI treatment. Since galactose feeding has little impact on levels of CNTF mRNA, these observations suggest that deficits in CNTF-like bioactivity may result from a posttranscriptional modification of neurotrophic protein expression or turnover. Unlike other functional and structural disorders in galactose neuropathy, factors other than polyol accumulation may contribute to the deficit in CNTF-like bioactivity.


Perivascular oxygen tensions in a transplantable mammary tumor growing in a dorsal flap window chamber.
Fischer 344 rats with R3230 Ac mammary carcinomas implanted in dorsal flap window chambers served as a model to obtain measurements of perivascular and stromal oxygen tension in normal and tumor tissues using Whalen recessed-tip microelectrodes (3- to 6-microns tip). Perivascular measurements were made adjacent to vessels with continuous blood flow. Thus the measurements and models provided are reflective of conditions leading to chronic hypoxia. Perivascular oxygen tensions averaged 72 +/- 13 mmHg in normal tissue vessels adjacent to tumor, 26 +/- 5 mmHg in tumor periphery, and 12 +/- 3 mmHg in tumor central vessels. There was a significant trend toward lower perivascular oxygen tensions in the tumor center (Kruskal-Wallis test, P = 0.002). A similar tendency was seen with a limited number of stromal measurements. Krogh cylinder models, which incorporate these data for perivascular oxygen tension, along with morphometric data obtained from the same tumor model suggest that hypoxic regions will exist between tumor vessels in the tumor center unless O2 consumption rates are well below 0.6 ml/100 g/min. The low perivascular measurements observed near the tumor center combined with the theoretical considerations suggest, for this model at least, that tissue oxygenation may best be improved by increasing red cell velocity and input pO2 and reducing oxygen consumption. The low perivascular oxygen tensions observed near the center also suggest that conditions conducive to increased red cell rigidity exist, that drugs which can decrease red cell rigidity could improve tumor blood flow and oxygenation, and that the endothelium of those vessels may be susceptible to hypoxia-reoxygenation injury.


PLASMODIUM BERGHEI: CYCLICAL TRANSMISSIONS BY EXPERIMENTALLY INFECTED ANOPHELES QUADRIMACULATUS.
A number of strains of Plasmodium berghei were isolated from sporozoites of Anopheles dureni. Laboratory-bred Anopheles quadrimaculatus fed on carriers of the newly isolated strains showed overwhelming midgut infections and moderate or mild salivary gland infections. Successive cyclic transmissions by the bite of experimentally infected A. quadrimaculatus in laboratorybred tree rats (Thamnomys surdaster) were carried out.


Motives for participating in a web-based nutrition cohort according to sociodemographic, lifestyle, and health characteristics: the NutriNet-Santé cohort study.
In traditional epidemiological studies, participants are likely motivated by perceived benefits, feelings of accomplishment, and belonging. No study has explored motives for participation in a Web-based cohort and the associated participant characteristics, although such information is useful for enhancing recruitment and improving cohort retention. We aimed to evaluate the relationships between motives for participation and sociodemographic, health, and lifestyle characteristics of participants in the NutriNet-Santé Web-based cohort, designed to identify nutritional risk or protective factors for chronic diseases. The motives for participation were assessed using a specifically developed questionnaire administered approximately 2 years after baseline. A total of 6352 completed the motives questionnaire (43.34%, 6352/15,000 randomly invited cohort participants). We studied the associations between motives (dependent variables) and individual characteristics with multivariate multinomial logistic regression models providing odds ratios and 95% confidence intervals. In total, 46.45% (2951/6352) of participants reported that they would not have enrolled if the study had not been conducted on the Internet, whereas 28.75% (1826/6352) were not sure. Men (OR 1.21, 95% CI 1.04-1.42), individuals aged 26-35 years (OR 1.51, 95% CI 1.20-1.91), and obese participants (OR 1.30, 95% CI 1.02-1.65) were more inclined to be motivated by the Internet aspect. Compared with younger adults and managerial staff, individuals >55 years (OR 0.60, 95% CI 0.48-0.45) and employees/manual workers were less likely motivated by the Internet aspect (OR 0.77, 95% CI 0.63-0.92). Regarding reasons for participation, 61.37% (3898/6352) reported participating to help advance public health research on chronic disease prevention; 22.24% (1413/6352) to help advance nutrition-focused research; 6.89% (438/6352) in response to the call from the media, after being encouraged by a close friend/associate, or a medical provider. Individuals >45 years (vs younger participants) were more likely (OR 1.62, 95% CI 1.07-2.47), whereas overweight and obese participants (vs nonobese participants) were less likely to participate in the study for reasons related to helping public health research on chronic disease prevention (OR 0.72, 95% CI 0.58-0.89; OR 0.62, 95% CI 0.46-0.84; respectively). Exclusive public funding of the study was important for 67.02% (4257/6352) of the participants. Men (OR 1.37, 95% CI 1.17-1.61) and persons >55 years (OR 1.97, 95% CI 1.57-2.47) were more likely to consider the exclusive public funding as very important. The use of the Internet, the willingness to help advance public health research, and the study being publicly funded were key motives for participating in the Web-based NutriNet-Santé cohort. These motives differed by sociodemographic profile and obesity, yet were not associated with lifestyle or health status. These findings can help improve the retention strategies in Web-based cohorts, particularly during decisive study periods when principal exposure information is collected.


Audit of the Health Service Executive orthodontic referral pathway between 2009 and 2011 in the Dublin Mid-Leinster region.
An audit was undertaken in 2009 to determine the success of the new national orthodontic referral protocol introduced to the Health Service Executive (HSE) in 2007 and operated in the Dublin Mid-Leinster HSE region. It was repeated in 2011 to determine if the HSE austerity measures have had a bearing on the orthodontic service performance in the Dublin Mid-Leinster HSE region. The audit also measured the success of referring practitioners in identifying the correct Index of Orthodontic Treatment Need (IOTN) classification of the patient. In the 2011 audit, the figures were broken down to identify the occlusal variables that caused dental practitioners most difficulties in identification. The audit demonstrates a good referral to assessment timeframe in 2009 (85-80% compliance for IOTN 5 and 4 within three to six months, respectively), which deteriorates significantly in 2011 (26-4% for IOTN 5 and 4 within three to six months, respectively). The ability of dentists to identify the correct IOTN classification was better in 2009 (60% correct) compared to 2011 (51% correct), but both figures fell below the audit standard of 75% of referrals with correct IOTN classifications. The IOTN occlusal dental health components most readily identified by referring practitioners and meeting audit standards were 5a (overjet >9mm), 5i (impacted teeth) and 5h (extensive hypodontia). The remaining occlusal dental health components in the HSE IOTN fell below the audit standard. The audit clearly identifies a requirement for a continued educational effort to maintain the HSE IOTN skill base in primary care, and a need for additional resources to manage the demand for orthodontic assessments.


Two-Tactor Vibrotactile Navigation Information for the Blind: Directional Resolution and Intuitive Interpretation.
Lacking vision, blind people have to exploit other senses for navigation. Using the tactile rather than the auditory sense avoids masking important environmental information. Directional information is particularly important and traditionally conveyed through an array of tactors, each coding one direction. Here, we present a different approach to represent arbitrary directions with only two tactors. We tested intuitiveness, plasticity, and variability of direction perception in a behavioral experiment in 33 seeing participants.


Correction of radiation sensitivity in ataxia telangiectasia cells by a truncated I kappa B-alpha.
Cells from patients with ataxia telangiectasia (AT) are hypersensitive to ionizing radiation and are defective in the regulation of DNA synthesis. A complementary DNA that corrects the radiation sensitivity and DNA synthesis defects in fibroblasts from an AT group D patient was isolated by expression cloning and shown to encode a truncated form of I kappa B-alpha, an inhibitor of the nuclear factor kappa B (NF-kappa B) transcriptional activator. The parental AT fibroblasts expressed large amounts of the I kappa B-alpha transcript and showed constitutive activation of NF-kappa B. The AT fibroblasts transfected with the truncated I kappa B-alpha expressed normal amounts of the I kappa B-alpha transcript and showed regulated activation of NF-kappa B. These results suggest that aberrant regulation of NF-kappa B and I kappa B-alpha contribute to the cellular defect in AT.


Colib'read on galaxy: a tools suite dedicated to biological information extraction from raw NGS reads.
With next-generation sequencing (NGS) technologies, the life sciences face a deluge of raw data. Classical analysis processes for such data often begin with an assembly step, needing large amounts of computing resources, and potentially removing or modifying parts of the biological information contained in the data. Our approach proposes to focus directly on biological questions, by considering raw unassembled NGS data, through a suite of six command-line tools. Dedicated to 'whole-genome assembly-free' treatments, the Colib'read tools suite uses optimized algorithms for various analyses of NGS datasets, such as variant calling or read set comparisons. Based on the use of a de Bruijn graph and bloom filter, such analyses can be performed in a few hours, using small amounts of memory. Applications using real data demonstrate the good accuracy of these tools compared to classical approaches. To facilitate data analysis and tools dissemination, we developed Galaxy tools and tool shed repositories. With the Colib'read Galaxy tools suite, we enable a broad range of life scientists to analyze raw NGS data. More importantly, our approach allows the maximum biological information to be retained in the data, and uses a very low memory footprint.


Chinese in west Malaysia: the geography of beta thalassaemia mutations.
The overseas Chinese in West Malaysia are almost exclusively from the south-eastern provinces of China-Kwangtung, Fukien, and Kwangsi. To institute a comprehensive thalassaemia control programme for this region we have characterised the beta thalassaemia mutations in 16 Chinese patients from West Malaysia: 4 beta thalassaemia mutations were seen: a) an A----G substitution in the TATA box [-28 base pairs (bp)], an A----T substitution in codon 17 [17 A----T], c) a 4 base pairs - TCTT deletion in codon 41-42 [frameshift mutation (FSC 41-42)], and d) a C----T substitution at the second intervening sequence (IVS 11) position 654. Similar mutations have been described in patients from the south-eastern provinces of China. The delineation of the specific mutations present will enable effective prenatal diagnosis for beta thalassaemia of ethnic Chinese in West Malaysia to be instituted.


Linoleic acid esterified in low density lipoprotein serves as substrate for increased arachidonic acid synthesis in differentiating monocytic cells.
The cellular metabolism of albumin- and lipoprotein-bound 18:2(n - 6) during monocytic differentiation was examined in the human premonocytic U937 and Mono Mac 6 cells. Differentiation for 72 h of U937 cells with retinoic acid (RA, 1 microM) or 1,25-(OH)2-vitamin D3 (1,25-D3, 10 nM) and of Mono Mac 6 cells with RA (1 microM) or lipopolysaccharide (LPS, 10 ng/ml) increased the desaturation and elongation of [1-14C]18:2(n - 6) to [1-14C]20:4(n - 6). In undifferentiated U937 and Mono Mac 6 cells, incubations with human LDL (100 micrograms/ml, 18 h) resulted in a 2.5-fold increase in 18:2(n - 6) levels in the cellular phospholipids. Differentiation of U937 cells with RA or or of Mono Mac 6 cells with LPS prior to LDL addition. Significantly reduced 18:2(n - 6) and elevated 20:4(n - 6) levels in cellular phospholipids. This increase in 20:4(n - 6) was likely not due to an increased incorporation of preformed 20:4(n - 6) esterified in LDL, as the receptor-specific degradation of [125I]LDL was reduced in both the RA-treated U937 and LPS-treated Mono Mac 6 cells. In U937 cells incubated with [1-14C]18:2(n - 6), the synthesis of TXB2, PGE2 and HHT could be detected after differentiation with RA. suggesting the availability of [1-14C]20:4(n - 6), derived from [1-14C]18:2(n - 6), for cyclooxygenase metabolism. Our results show that the conversion of 18:2(n - 6) to 20:4(n - 6) increases during monocyte differentiation. The 18:2(n - 6) supplied to the cells via the receptor-mediated uptake of LDL was utilized as substrate for the increased 20:4(n - 6) synthesis.


Surface modulation of silicon nitride ceramics for orthopaedic applications.
Silicon nitride (Si3N4) has a distinctive combination of material properties such as high strength and fracture toughness, inherent phase stability, scratch resistance, low wear, biocompatibility, hydrophilic behavior, excellent radiographic imaging and resistance to bacterial adhesion, all of which make it an attractive choice for orthopaedic implants. Unlike oxide ceramics, the surface chemistry and topography of Si3N4 can be engineered to address potential in vivo needs. Morphologically, it can be manufactured to have an ultra-smooth or highly fibrous surface structure. Its chemistry can be varied from that of a silica-like surface to one which is predominately comprised of silicon-amines. In the present study, a Si3N4 bioceramic was subjected to thermal, chemical, and mechanical treatments in order to induce changes in surface composition and features. The treatments included grinding and polishing, etching in aqueous hydrofluoric acid, and heating in nitrogen or air. The treated surfaces were characterized using a variety of microscopy techniques to assess morphology. Surface chemistry and phase composition were determined using X-ray photoelectron and Raman spectroscopy, respectively. Streaming potential measurements evaluated surface charging, and sessile water drop techniques assessed wetting behavior. These treatments yielded significant differences in surface properties with isoelectric points ranging from 2 to 5.6, and moderate to extremely hydrophilic water contact angles from ∼65° to ∼8°. This work provides a basis for future in vitro and in vivo studies which will examine the effects of these treatments on important orthopaedic properties such as friction, wear, protein adsorption, bacteriostasis and osseointegration. Silicon nitride (Si3N4) exhibits a unique combination of bulk mechanical and surface chemical properties that make it an ideal biomaterial for orthopaedic implants. It is already being used for interbody spinal fusion cages and is being developed for total joint arthroplasty. Its surface texture and chemistry are both highly tunable, yielding physicochemical combinations that may lead to enhanced osseointegration and bacterial resistance without compromising bulk mechanical properties. This study demonstrates the ease with which significant changes to Si3N4's surface phase composition, charging, and wetting behavior can be induced, and represents an initial step towards a mechanistic understanding of the interaction between implant surfaces and the biologic environment.


CARD14 expression in dermal endothelial cells in psoriasis.
Mutations in the caspase recruitment domain, family member 14 (CARD14) gene have recently been described in psoriasis patients, and explain the psoriasis susceptibility locus 2 (PSORS2). CARD14 is a scaffolding protein that regulates NF-κB activation, and psoriasis-associated CARD14 mutations lead to enhanced NF-κB signaling. CARD14 is expressed mainly in epidermal keratinocytes, but also in unidentified dermal cells. In this manuscript, the identity of the dermal cell types expressing CARD14, as well the potential functional consequence of overactive CARD14 in these dermal cell types, was determined. Using two-color immunofluorescence, dermal CARD14 did not co-localize with T-cells, dendritic cells, or macrophages. However, dermal CARD14 did highly co-localize with CD31(+) endothelial cells (ECs). CARD14 was also expressed non-dermal endothelial cells, such as aortic endothelial cells, which may indicate a role of CARD14(+)ECs in the systemic inflammation and cardiovascular comorbidities associated with psoriasis. Additionally, phosphorylated NF-κB was found in psoriatic CARD14(+) CD31(+) ECs, demonstrating this pathway is active in dermal ECs in psoriasis. Transfection of dermal ECs with psoriasis-associated CARD14 mutations resulted in increased expression of several chemokines, including CXCL10, IL-8, and CCL2. These results provide preliminary evidence that CARD14 expression in ECs may contribute to psoriasis through increased expression of chemokines and facilitating recruitment of immune cells into skin.


Low-density lipoprotein receptor-related protein 1 promotes cancer cell migration and invasion by inducing the expression of matrix metalloproteinases 2 and 9.
The low-density lipoprotein receptor-related protein 1 (LRP1) is a multifunctional endocytic receptor involved in the metabolism of various extracellular ligands, including proteinases, that play critical roles in tumor invasion. Although several studies have shown an increased expression of LRP1 in cancer cells, its function in tumor development and progression remains largely unclear. Here, we reveal a novel mechanism by which LRP1 induces the expression of matrix metalloproteinase 2 (MMP2) and MMP9 and thereby promotes the migration and invasion of human glioblastoma U87 cells. Knockdown of LRP1 expression greatly decreased U87 cell migration and invasion, which was rescued by the forced expression of a functional LRP1 minireceptor. Inhibition of ligand binding to LRP1 by a specific antagonist, receptor-associated protein, also led to reduced cancer cell migration and invasion. Because MMPs play critical roles in cancer cell migration and invasion, we examined the expression of several MMPs and found that the expression of functional MMP2 and MMP9 was selectively decreased in LRP1 knockdown cells. More importantly, decreased cell migration and invasion of LRP1 knockdown cells were completely rescued by exogenous expression of MMP2 or MMP9, suggesting that these MMPs are likely downstream targets of LRP1-mediated signaling. We further show that the level of phosphorylated extracellular signal-regulated kinase (ERK) was significantly decreased in LRP1-silenced cells, suggesting that ERK is a potential mediator of LRP1-regulated MMP2 and MMP9 expression in U87 cells. Together, our data strongly suggest that LRP1 promotes glioblastoma cell migration and invasion by regulating the expression and function of MMP2 and MMP9 perhaps via an ERK-dependent signaling pathway.


Lessons from malaria control to help meet the rising challenge of dengue.
Achievements in malaria control could inform efforts to control the increasing global burden of dengue. Better methods for quantifying dengue endemicity-equivalent to parasite prevalence surveys and endemicity mapping used for malaria-would help target resources, monitor progress, and advocate for investment in dengue prevention. Success in controlling malaria has been attributed to widespread implementation of interventions with proven efficacy. An improved evidence base is needed for large-scale delivery of existing and novel interventions for vector control, alongside continued investment in dengue drug and vaccine development. Control of dengue is unlikely to be achieved without coordinated international financial and technical support for national programmes, which has proven effective in reducing the global burden of malaria.


Food color, flavor, and conditioned avoidance among red-winged blackbirds.
The relationship between food flavors and postingestive feedback enables mammalian herbivores to procure nutrients and avoid toxins within ever-changing environments. We conducted four experiments with red-winged blackbirds (Agelaius phoeniceus) in captivity to determine the relative roles of color and flavor cues paired with negative postingestive feedback. We first conducted baseline tests to assess preferences for colors and flavors. All blackbirds preferred red- to blue-colored food, and they preferred umami- (l-alanine) flavored to bitter/astringent food (tannic acid). We observed no difference in consumption of salty (NaCl) vs sour (citric acid) foods during baseline tests (i.e., neutral flavors). We then conditioned experimentally naïve blackbirds with intraperitoneal injections of lithium chloride (LiCl) to avoid food treated with red and l-alanine, or red and NaCl (n=30 birds per conditioning group). Subsequent to conditioning with LiCl, three test groups were established from each conditioned group to evaluate color and flavor preferences, and preferences for novel color-flavor pairings (e.g., red/tannic acid vs blue/l-alanine). Blackbirds avoided red and salty food throughout the 4-day test. Avoidance conditioned with LiCl extinguished for preferred flavors, but not for colors, of food. Conditioning affected indifference for the otherwise preferred flavor and avoidance for the otherwise neutral flavor. Relative to the neutral-flavor conditioning group, the group conditioned with a preferred flavor exhibited stronger conditioned avoidance of colored food. Unlike conditioned flavor avoidance, birds were conditioned to avoid red food only when blue food was made familiar prior to conditioning. Collectively, these results illustrate that blackbirds used affective processes (flavor-feedback relationships) to shift preference for both novel and familiar flavors, and cognitive associations (colors) to avoid food, subsequent to toxin exposure. We discuss the opportunities afforded by affective and cognitive processing for reducing agricultural damage caused by blackbirds.


Metal-coordination: Using one of nature's tricks to control soft material mechanics.
Growing evidence supports a critical role of dynamic metal-coordination crosslinking in soft biological material properties such as self-healing and underwater adhesion1. Using bio-inspired metal-coordinating polymers, initial efforts to mimic these properties have shown promise2. Here we demonstrate how bio-inspired aqueous polymer network mechanics can be easily controlled via metal-coordination crosslink dynamics; metal ion-based crosslink stability control allows aqueous polymer network relaxation times to be finely tuned over several orders of magnitude. In addition to further biological material insights, our demonstration of this compositional scaling mechanism should provide inspiration for new polymer material property-control designs.


Betaine Alleviates Cognitive Deficits in Diabetic Rats via PI3K/Akt Signaling Pathway Regulation.
Diabetes mellitus is a metabolic disease which also causes cognitive deficits. Betaine (N,N,N-trimethylglycine), also known as trimethylglycine, has been shown to ameliorate diabetic symptoms in diabetic animals and improve cognitive ability in Alzheimer disease (AD) animals. However, the effects of betaine on cognitive deficits in diabetic animals have not been described yet. Therefore, in the current study, the effects of betaine on cognition in diabetic rats were evaluated. We established a diabetic rat model by injecting streptozotocin (STZ) into rats and administrated betaine to these diabetic rats. We monitored the metabolism index, and glucose and insulin levels in blood and cerebrospinal fluid. We measured inflammatory cytokine levels, including TNF-α, IL-1β, and IL-6, in serum and hippocampus. We also monitored oxidative stress in the hippocampus by measuring malondialdehyde (MDA) level and superoxide dismutase (SOD) activity. We measured the learning and memory ability of diabetic rats using the Morris water and Y maze tests and tested the phosphatidylinositol 3-kinase (PI3K)/Akt activation and p-mTOR level in the hippocampus. Betaine improved glucose metabolism and suppressed the production of inflammatory cytokines, including TNF-α, IL-1β, and IL-6. Also, betaine decreased MDA concentration and increased SOD activity in the hippocampus of diabetic rats. Betaine ameliorated cognitive deficits in diabetic rats, and it promoted PI3K expression and Akt activation and decreased p-mTOR expression. Betaine alleviates cognitive deficits in STZ-induced diabetic rats via regulating the PI3K/Akt signaling pathway.


Erysipelothrix rhusiopathiae contamination in the poultry house environment during erysipelas outbreaks in organic laying hen flocks.
This study investigated organic laying hen farms for the presence of Erysipelothrix rhusiopathiae in the house environment and from potential carriers (i.e. insects and mice) during ongoing erysipelas outbreaks, and compared the obtained isolates with those from laying hens. The samples were investigated by selective culture followed by species-specific polymerase chain reaction on cultures. E. rhusiopathiae was isolated from the spleen, jejunal contents, manure, dust and swabs from water nipples. Three more samples from the house environment tested positive by polymerase chain reaction compared with selective culture alone. Selected isolates were investigated by pulsed-field gel electrophoresis (PFGE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). One farm was represented by isolates from laying hens only, and one of these isolates differed in one PFGE band from the others. Different banding patterns were observed for isolates from laying hens and manure on one farm. On the remaining two farms, the isolates from the house environment and laying hens were identical but differed between farms. Outbreaks reoccurred in the next flock on two of the farms, and different PFGE types were isolated from consecutive flocks. Our results suggest an external source of infection, which would explain the previously reported increased risk of outbreaks in free-range flocks. Contaminated manure and dust may represent sources of transmission. For the isolates, MALDI-TOF MS and biochemical typing results were in agreement but, since the type strain of Erysipelothrix tonsillarum was typed as E. rhusiopathiae using MALDI-TOF MS, further studies into this method are needed.


Molecular and epigenetic analysis of the fragile histidine triad tumour suppressor gene in equine sarcoids.
Sarcoids are peculiar equine benign tumours. Their onset is associated with Bovine Papillomavirus type -1 or -2 (BPV-1/2) infection. Little is known about the molecular interplay between viral infection and neoplastic transformation. The data regarding papillomavirus infections in human species show the inactivation of a number of tumour suppressor genes as basic mechanism of transformation. In this study the putative role of the tumour suppressor gene Fragile Histidine Triad (FHIT) in sarcoid tumour was investigated in different experimental models. The expression of the oncosuppressor protein was assessed in normal and sarcoid cells and tissue. Nine paraffin embedded sarcoids and sarcoid derived cell lines were analysed for the expression of FHIT protein by immunohistochemistry, immunofluorescence techniques and western blotting. These analyses revealed the absence of signal in seven out of nine sarcoids. The two sarcoid derived cell lines too showed a reduced signal of the protein. To investigate the causes of the altered protein expression, the samples were analysed for the DNA methylation profile of the CpG island associated with the FHIT promoter. The analysis of the 32 CpGs encompassing the region of interest showed no significative differential methylation profile between pathological tissues and cell lines and their normal counterparts. This study represent a further evidence of the role of a tumour suppressor gene in equine sarcoids and approaches the epigenetic regulation in this well known equine neoplasm. The data obtained in sarcoid tissues and sarcoid derived cell lines suggest that also in horse, as in humans, there is a possible involvement of the tumour suppressor FHIT gene in BPV induced tumours. DNA methylation seems not to be involved in the gene expression alteration. Further studies are needed to understand the basic molecular mechanisms involved in reduced FHIT expression.


Distal internal iliac artery embolization: a procedure to avoid.
Internal iliac artery (IIA) coil embolization as an adjunct to endovascular stent grafting (ESG) is common practice for treating abdominal aortic aneurysm (AAA) in patients with a substantially enlarged common iliac artery requiring extension of the stent-graft limb into the external iliac artery. The literature describing pelvic ischemia in association with IIA coil embolization contains conflicting reports of symptom severity. We studied IIA occlusion outcome as a function of coil placement in the IIA. From August 1997 to March 2002, 20 patients with AAA underwent ESG with unilateral IIA coil embolization. Coils were placed proximal to the first branch of the IIA in 8 patients and distal to the first branch in 12 patients. Symptoms of pelvic ischemia and mid-term outcome were studied. Patients included 18 men and 2 women with mean age of 70(1/2) years (range, 53-86 years). Mean diameter of AAA was 54.4 mm (range, 38-80 mm), and of common iliac artery was 24.2 mm (range, 15-48 mm). Ten patients (50%) had new onset of symptoms of pelvic ischemia after endograft procedures: 1 of 8 patients (13%) with proximal IIA embolization had buttock claudication, and 9 of 12 patients (75%) with distal IIA embolization had pelvic ischemic symptoms, including buttock claudication in 8 and impotence in 1 (P =.02, Fisher exact test). No colonic ischemia occurred in this series. At 12-month follow-up, 4 patients with distal IIA embolization were symptom-free. At further follow-up to 24 months, 4 patients remained significantly limited with symptoms of claudication. A significantly higher incidence of symptoms of pelvic ischemia occurred with more distal placement of coils for IIA embolization. Failure to control for extent of coil placement may account for the apparently conflicting results in published studies. IIA coil embolization should be performed as proximal as possible to prevent interference with pelvic collateral circulation.


Recombinant protein expression by targeting pre-selected chromosomal loci.
Recombinant protein expression in mammalian cells is mostly achieved by stable integration of transgenes into the chromosomal DNA of established cell lines. The chromosomal surroundings have strong influences on the expression of transgenes. The exploitation of defined loci by targeting expression constructs with different regulatory elements is an approach to design high level expression systems. Further, this allows to evaluate the impact of chromosomal surroundings on distinct vector constructs. We explored antibody expression upon targeting diverse expression constructs into previously tagged loci in CHO-K1 and HEK293 cells that exhibit high reporter gene expression. These loci were selected by random transfer of reporter cassettes and subsequent screening. Both, retroviral infection and plasmid transfection with eGFP or antibody expression cassettes were employed for tagging. The tagged cell clones were screened for expression and single copy integration. Cell clones producing > 20 pg/cell in 24 hours could be identified. Selected integration sites that had been flanked with heterologous recombinase target sites (FRTs) were targeted by Flp recombinase mediated cassette exchange (RMCE). The results give proof of principle for consistent protein expression upon RMCE. Upon targeting antibody expression cassettes 90-100% of all resulting cell clones showed correct integration. Antibody production was found to be highly consistent within the individual cell clones as expected from their isogenic nature. However, the nature and orientation of expression control elements revealed to be critical. The impact of different promoters was examined with the tag-and-targeting approach. For each of the chosen promoters high expression sites were identified. However, each site supported the chosen promoters to a different extent, indicating that the strength of a particular promoter is dominantly defined by its chromosomal context. RMCE provides a powerful method to specifically design vectors for optimized gene expression with high accuracy. Upon considering the specific requirements of chromosomal sites this method provides a unique tool to exploit such sites for predictable expression of biotechnologically relevant proteins such as antibodies.


Preventive Effect of Glycyrrhiza Glabra Extract on Oral Mucositis in Patients Under Head and Neck Radiotherapy: A Randomized Clinical Trial.
About two-thirds of cancer patients undergo radiotherapy. Oral mucositis represents a major complication of radiotherapy, causing morbidity and mortality and decreasing the quality of life of patients. This study aimed to assess the preventive effect of Glycyrrhiza aqueous extract on oral mucositis in cancer patients under head and neck radiotherapy. In this double-blind clinical trial, 37 head and neck cancer patients were divided into intervention (n=19) group receiving Glycyrrhiza aqueous extract and control (n=18) group receiving placebo. Patients in the test group used Glycyrrhiza aqueous extract topically twice a day from the first day of starting radiotherapy until the end of the second week. Patients were examined in the first day of radiotherapy for any type of wound before treatment and those with oral ulcers before radiotherapy were excluded from the study. The grade of mucositis was determined using the classification by the World Health Organization. ANCOVA was performed to assess any difference between the two groups with regard to oral mucosal irritation and wound size after the intervention while controlling for the covariates such as sex and age. Significant differences were found in the maximum grade of mucositis and oral mucosal irritation between the intervention and control groups (P<0.001). This study showed that aqueous extract of Glycyrrhiza can be effective for decreasing the severity of oral mucositis in head and neck cancer patients undergoing radiotherapy.


Multiple Health Behavior Programs in School Settings: Strategies to Promote Transfer-of-Learning Through Life Skills Education.
Typically, schools implement health promotion programs that focus on a single behavioral domain. Multiple related health topics may be addressed using separate interventions, potentially producing overlap in program content. However, integrative approaches in health promotion have the potential to produce interventions capable of improving multiple health behaviors. In particular, more research is needed to identify the conditions and the factors that can promote the transfer of learning to broaden the target outcomes of health promotion programs. The present study aims to identify the characteristics of an evidence-based life skills education program that can facilitate the transfer of learning to different health behaviors not initially targeted by the program, and the strategies for achieving successful transfer. A two round Delphi method was used with a diverse group of 21 experts in health promotion, life skills education, and methods of pedagogy for early adolescent students. Questionnaires with open and closed-ended questions were administered on-line. Content qualitative analysis was run, integrating codes, subcategories, and categories of the two rounds of the study. Results showed strong consensus among experts about the potential for promoting the transfer of skills from one health domain to another. Many elements were identified as important factors that facilitate the transfer of learning. Strategies for successful transfer were related to teaching methods, educational approaches, and consistency with the broader school curriculum. Findings suggest that the successful transfer of learning to a new health domain requires that educators recognize its importance and explicitly designate it as an educational aim.


Association of Anesthesiologist Handovers With Short-term Outcomes for Patients Undergoing Cardiac Surgery.
Complete handover of anesthesia care to a second anesthesiologist has been demonstrated to be associated with worse short-term adverse outcomes among cardiac surgery patients, but little information from multi-institutional studies is available. New York's cardiac surgery registry was used to identify patients who underwent cardiac surgery in New York between 2010 and 2016 with and without complete handovers of anesthesia care. A retrospective observational study with inverse probability treatment weighting (IPTW) based on the propensity score was used to adjust for differences in preoperative patient characteristics while comparing differences in the primary outcome (in-hospital/30 day mortality), major complications in the index admission or within 30 days of the index surgery, readmissions within 30 days, and length of stay. A total of 8.5% of the 103,102 cardiac surgery procedures involved complete handovers. After adjustment, there was a difference between patients with and without handovers in the primary outcome (2.86% vs 2.48%, adjusted risk ratio [ARR] = 1.15 [1.01-1.31]). There was no difference in readmissions within 30 days (13.7% vs 14.4%, ARR = 0.95 [0.90-1.00]), and the differences in complications and length of stay were not clinically meaningful (adjusted differences of <10%). Cardiac surgery patients in New York who had complete anesthesia handovers experienced higher short-term mortality rates, but there were no meaningful differences in other outcomes. Unnecessary handovers should be carefully monitored.


Low-molecular-weight heparins or heparinoids versus standard unfractionated heparin for acute ischaemic stroke.
Low-molecular-weight heparins and heparinoids are anticoagulants that may be associated with lower risks of haemorrhage and more powerful antithrombotic effects than standard unfractionated heparin. This is an updated version of the original Cochrane review first published in Issue 1, 1995 and previously updated in Issue 2, 2005. To compare the effects of low-molecular-weight heparins or heparinoids with those of unfractionated heparin in people with acute, confirmed or presumed, ischaemic stroke. We searched the Cochrane Stroke Group Trials Register (last searched June 2007). In addition we searched the Cochrane Central Register of Controlled Trials (The Cochrane Library Issue 2, 2007), MEDLINE (1966 to June 2007) and EMBASE (1980 to June 2007). For previous versions of this review we searched MedStrategy (1995) and also contacted pharmaceutical companies. Randomised trials comparing heparinoids or low-molecular-weight heparins with standard unfractionated heparin in people with acute ischaemic stroke. We only included trials where treatment was started within 14 days of stroke onset. Two review authors independently selected studies for inclusion, assessed trial quality and extracted the data. Nine trials involving 3137 people were included. Four trials compared a heparinoid (danaparoid), four trials compared low-molecular-weight heparin (enoxaparin or certoparin), and one trial compared an unspecified low-molecular-weight heparin with standard unfractionated heparin. Allocation to low-molecular-weight heparin or heparinoid was associated with a significant reduction in the odds of deep vein thrombosis compared with standard unfractionated heparin (odds ratio (OR) 0.55, 95% confidence interval (CI) 0.44 to 0.70). However, the number of more major events (pulmonary embolism, death, intracranial or extracranial haemorrhage) was too small to provide a reliable estimate of the benefits and risks of low-molecular-weight heparins or heparinoids compared with standard unfractionated heparin for these, arguably more important, outcomes. Insufficient information was available to assess effects on recurrent stroke or functional outcome. Since the last version of this review none of the three new relevant studies with 2397 participants have provided additional information to change the conclusions. Treatment with a low-molecular-weight heparin or heparinoid after acute ischaemic stroke appears to decrease the occurrence of deep vein thrombosis compared with standard unfractionated heparin, but there are too few data to provide reliable information on their effects on other important outcomes, including death and intracranial haemorrhage.


[Efficacy assessment of Nutrilarm®, a per os omega-3 and omega-6 polyunsaturated essential fatty acid dietary formulation versus placebo in patients with bilateral treated moderate dry eye syndrome].
Inflammation is one of the main mechanisms common to all forms of dry eye. Since polyunsaturated acids are known to show biological anti-inflammatory properties, the aim of this study was to evaluate the efficacy of dietary n-6 and n-3 fatty acids in patients suffering from ocular dryness. One hundred and eighty-one patients diagnosed with bilateral moderate dry eye who were already treated with lachrymal substitutes were randomized in a double-blind international study to receive placebo or Nutrilarm(®) capsules (combination of omega-3 and omega-6), twice a day for 6 months. In all subjects, dryness feeling, overall subjective comfort, and ocular symptoms (burning, stinging, sandy and/or gritty sensation, light sensitivity, reflex tearing, and ocular fatigue) were evaluated at each visit. Furthermore, fluorescein tests (break-uptime and Oxford scheme) and lissamine green test were performed at each visit. The Schirmer test was performed at inclusion and after 6 months of treatment. After 6 months of supplementation with Nutrilarm(®), both the BUT scores and ocular fatigue were significantly improved when compared with placebo (P=0.036 and P=0.044, respectively). There was a trend in favor of Nutrilarm(®) in terms of the efficacy evaluated by the investigator (P=0.061). Fewer patients experienced a feeling of severe dryness with Nutrilarm(®) compared with placebo after 6 months of treatment (2.5 and 9.3%, respectively), but the difference was not statistically significant. Oral administration of a double supplementation dietary n-6 and n-3 fatty acids present an additional therapeutic advantage in patients suffering from ocular dryness who were already treated with lachrymal substitutes.


Development and application of an aerosol screening model for size-resolved urban aerosols.
Predictive models of vehicular ultrafine particles less than 0.1 microm in diameter (UFPs*) and other urban pollutants with high spatial and temporal variation are useful and important in applications such as (1) decision support for infrastructure projects, emissions controls, and transportation-mode shifts; (2) the interpretation and enhancement of observations (e.g., source apportionment, extrapolation, interpolation, and gap-filling in space and time); and (3) the generation of spatially and temporally resolved exposure estimates where monitoring is unfeasible. The objective of the current study was to develop, test, and apply the Aerosol Screening Model (ASM), a new physically based vehicular UFP model for use in near-road environments. The ASM simulates hourly average outdoor concentrations of roadway-derived aerosols and gases. Its distinguishing features include user-specified spatial resolution; use of the Weather Research and Forecasting (WRF) meteorologic model for winds estimates; use of a database of more than 100,000 road segments in the Los Angeles, California, region, including freeway ramps and local streets; and extensive testing against more than 9000 hours of observed particle concentrations at 11 sites. After initialization of air parcels at an upwind boundary, the model solves for vehicle emissions, dispersion, coagulation, and deposition using a Lagrangian modeling framework. The Lagrangian parcel of air is subdivided vertically (into 11 levels) and in the crosswind direction (into 3 parcels). It has overall dimensions of 10 m (downwind), 300 m (vertically), and 2.1 km (crosswind). The simulation is typically started 4 km upwind from the receptor, that is, the location at which the exposure is to be estimated. As parcels approach the receptor, depending on the user-specified resolution, step size is decreased, and crosswind resolution is enhanced through subdivision of parcels in the crosswind direction. Hourly concentrations and size distributions of aerosols were simulated for 11 sites in the Los Angeles area with large variations in proximal traffic and particle number concentrations (ranging from 6000 to 41,000/cm3). Observed data were from the 2005-2007 Harbor Community Monitoring Study (HCMS; Moore et al. 2009), in Long Beach, California, and the Coronary Health and Air Pollution Study (CHAPS; Delfino et al. 2008), in the Los Angeles area. Meteorologic fields were extracted from 1-km-resolution meteorologic simulations, and observed wind direction and speed were incorporated. Using on-road and tunnel measurements, size-resolved emission factors ranging from 1.4 x 10(15) to 16 x 10(15) particles/kg fuel were developed specifically for the ASM. Four separate size-resolved emissions were used. Traffic and emission factors were separately estimated for heavy-duty diesel and light-duty vehicles (LDV), and both cruise and acceleration emission factors were used. The light-duty cruise size-resolved number emission factor had a single prominent mode at 12 nm. The diesel cruise size-resolved number emission factor was bimodal, with a large mode at 16 nm and a secondary mode at around 100 nm. Emitted particles were assumed to be nonvolatile. Data on traffic activity came from a 2008 travel-demand model, supplemented by data on diurnal patterns. Simulated ambient number size distributions and number concentrations were compared to observations taking into account estimated losses from particle transmission efficiency in instrument inlet tubing. The skill of the model in predicting number concentrations and size distributions was mixed, with some promising prediction features and some other areas in need of substantial improvement. For long-term (-15-day) average concentrations, the variability from site to site could be modeled with a coefficient of determination (r2) of 0.76. Model underprediction was more common than overprediction. The average of the absolute normalized bias was 0.30; in other words, long-term mean particle concentrations at each site were on average predicted to within 30% of the measured values. Observed 24-hour number concentrations were simulated to within a factor of 1.6 on 48% of days at HCMS sites and 81% at CHAPS sites, lower than the original design goal of 90%. Extensive evaluation of hourly concentrations, diurnal patterns, sizedistributions, and directional patterns was performed. At two sites with heavy freeway and heavy-duty-vehicle (HDV) influences and extensive size-resolved measurements, the ASM made significant errors in the diurnal pattern, concentration, and mode position of the aerosol size distribution. Observations indicated a shift in concentrations and size distributions corresponding to the afternoon development of offshore wind at the HCMS sites. The model did not reproduce the changes in particles associated with this wind shift and suffered from overprediction for particles of less than 15 nm and underprediction for particles of between 15 and 500 nm, raising doubt about the applicability of the HDV emission factors and the model's assumptions that particles were nonvolatile. The model's temporal prediction skill at individual monitoring sites was variable; the index of agreement (IOA) for hourly values at single sites ranged from 0.30 to 0.56. The model's ability to reproduce diurnal patterns in aerosol concentrations was site dependent; midday underprediction as well as underprediction for particle sizes greater than 15 nm were typical errors. Despite some problems in model skill, the number of time periods and locations evaluated as well as the extent of our qualitative and quantitative evaluations versus physical measurements well exceeded other published size-resolved modeling efforts. As a trial of a typical application, the sensitivity of the concentrations at each receptor site to LDV traffic, HDV traffic, and various road classes was evaluated. The sensitivity of overall particle numbers to all types of traffic ranged from 0.87 at the site with the heaviest traffic to 0.28 at the site with the lightest traffic, meaning that a 1% reduction in traffic could yield a reduction in particle number of 0.87% to 0.28%. Key conclusions and implications of the study are the following: 1. That variable-resolution (down to 10 m) modeling in a relatively simple framework is feasible and can support most of the applications mentioned above; 2. That model improvements will be required for some applications, especially in the areas of the HDV emission factor and the parameterization of meteorologic dispersion; 3. That particle loss from instrument transmission efficiency can be significant for particles smaller than 50 nm, and especially significant for particles smaller than 20 nm. In cases where loss corrections are not accounted for, or are inaccurate, this loss can cause disagreements in observation-model and observation-observation comparisons. 4. That LDV traffic exposures likely exceed HDV traffic exposures in some locations; 5. That variable step size and adaptive parcel width are critical to balancing computational efficiency and resolution; and 6. That the effects of roadways on air quality depend on both traffic volume and distance--in other words, low traffic volumes at close proximity need to be considered in health and planning studies just as much as do high traffic volumes at distances up to several kilometers. Future improvements to the model have been identified. They include improved emission factors; integration with the U.S. Environmental Protection Agency (EPA) Motor Vehicle Emission Simulator (MOVES) model; nesting with three-dimensional (3D) Eulerian models such as the Community Multi-scale Air Quality (CMAQ) model; increased emission dependence on acceleration, load, grade, and speed as well as evaporation and condensation of semivolatile aerosol species; and modeling of carbon dioxide (CO2) as an on-road and near-road dilution tracer. In addition, comparison with other statistically and physically based models would be highly beneficial.


Short-term, but not long-term, increased daytime workload leads to decreased night-time energetics in a free-living song bird.
Reproduction is energetically expensive and to obtain sufficient energy, animals can either alter their metabolic system over time to increase energy intake (increased-intake hypothesis) or reallocate energy from maintenance processes (compensation hypothesis). The first hypothesis predicts a positive relationship between basal metabolic rate (BMR) and energy expenditure (DEE) because of the higher energy demands of the metabolic system at rest. The second hypothesis predicts a trade-off between different body functions, with a reduction of the BMR as a way to compensate for increased daytime energetic expenditure. We experimentally manipulated the workload of wild pied flycatchers by adding or removing chicks when chicks were 2 and 11 days old. We then measured the feeding frequency (FF), DEE and BMR at day 11, allowing us to assess both short- and long-term effects of increased workload. The manipulation at day 2 caused an increase in FF when broods were enlarged, but no response in DEE or BMR, while the manipulation at day 11 caused an increase in FF, no change in DEE and a decrease in BMR in birds with more chicks. Our results suggest that pied flycatchers adjust their workload but that this does not lead to a higher BMR at night (no support for the increased-intake hypothesis). In the short term, we found that birds reallocate energy with a consequent reduction of BMR (evidence for the compensation hypothesis). Birds thus resort to short-term strategies to increase energy expenditure, which could explain why energy expenditure and hard work are not always correlated in birds.


Establishing an antimicrobial stewardship program.
Hospitals are faced with increasing challenges of antibiotic-resistant organisms and rising antimicrobial costs despite widespread attention to infection prevention and control measures. Government, professional organizations and accreditation bodies are all signalling an urgent need for the establishment of programs in hospitals to address antibiotic misuse. Although variations of such "antimicrobial stewardship programs" have been functioning in Canada for some time, a formal approach using change management and quality improvement principles has largely been lacking. We describe how we have established such a program in a teaching hospital, modelled on John Kotter's eight steps of leading change.


Angiotensinogen gene silencing reduces markers of lipid accumulation and inflammation in cultured adipocytes.
Inflammatory adipokines secreted from adipose tissue are major contributors to obesity-associated inflammation and other metabolic dysfunctions. We and others have recently documented the contribution of adipose tissue renin-angiotensin system to the pathogenesis of obesity, inflammation, and insulin resistance. We hypothesized that adipocyte-derived angiotensinogen (Agt) plays a critical role in adipogenesis and/or lipogenesis as well as inflammation. This was tested using 3T3-L1 adipocytes, stably transfected with Agt-shRNA or scrambled Sc-shRNA as a control. Transfected preadipocytes were differentiated and used to investigate the role of adipose Agt through microarray and PCR analyses and adipokine profiling. As expected, Agt gene silencing significantly reduced the expression of Agt and its hormone product angiotensin II (Ang II), as well as lipid accumulation in 3T3-L1 adipocytes. Microarray studies identified several genes involved in lipid metabolism and inflammatory pathways which were down-regulated by Agt gene inactivation, such as glycerol-3-phosphate dehydrogenase 1 (Gpd1), serum amyloid A 3 (Saa3), nucleotide-binding oligomerization domain containing 1 (Nod1), and signal transducer and activator of transcription 1 (Stat1). Mouse adipogenesis PCR arrays revealed lower expression levels of adipogenic/lipogenic genes such as peroxisome proliferator activated receptor gamma (PPARγ), sterol regulatory element binding transcription factor 1 (Srebf1), adipogenin (Adig), and fatty acid binding protein 4 (Fabp4). Further, silencing of Agt gene significantly lowered expression of pro-inflammatory adipokines including interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and monocyte chemotactic protein-1 (MCP-1). In conclusion, this study directly demonstrates critical effects of Agt in adipocyte metabolism and inflammation and further support a potential role for adipose Agt in the pathogenesis of obesity-associated metabolic alterations.