The impact of a decision aid about heart disease prevention on patients' discussions with their doctor and their plans for prevention: a pilot randomized trial.
Low utilization of effective coronary heart disease (CHD) prevention strategies may be due to many factors, but chief among them is the lack of patient involvement in prevention decisions. We undertook this study to test the effectiveness of an individually-tailored, computerized decision aid about CHD on patients' discussions with their doctor and their plans for CHD prevention. We conducted a pilot randomized trial in a convenience sample of adults with no previous history of cardiovascular disease to test the effectiveness of an individually-tailored, computerized decision aid about CHD prevention against a risk factor list that patients could present to their doctor. We enrolled 75 adults. Mean age was 53. 59% were female, 73% white, and 23% African-American. 66% had some college education. 43% had a 10-year CHD risk of 0-5%, 25% a risk of 6-10%, 24% a risk of 11-20%, and 5% a risk of > 20%. 78% had at least one option to reduce their CHD risk, but only 45% accurately identified the strategies best supported by evidence. 41 patients received the decision aid, 34 received usual care. In unadjusted analysis, the decision aid increased the proportion of patients who discussed CHD risk reduction with their doctor from 24% to 40% (absolute difference 16%; 95% CI -4% to +37%) and increased the proportion who had a specific plan to reduce their risk from 24% to 37% (absolute difference 13%; 95% CI -7% to +34%). In pre-post testing, the decision aid also appeared to increase the proportion of patients with plans to intervene on their CHD risk (absolute increase ranging from 21% to 47% for planned medication use and 5% to 16% for planned behavioral interventions). Our study confirms patients' limited knowledge about their CHD risk and effective risk reduction options and provides preliminary evidence that an individually-tailored decision aid about CHD prevention might be expected to increase patients' discussions about CHD prevention with their doctor and their plans for CHD risk reduction. These findings should be replicated in studies with a larger sample size and patients at overall higher risk of CHD.


Synergistic Effect of Colistin and Rifampin Against Multidrug Resistant Acinetobacter baumannii: A Systematic Review and Meta-Analysis.
The existence of infections caused by multidrug resistant (MDR) Acinetobacter baumannii is a growing problem because of the difficulty to treat them. We examined the published literature and focused our analysis on the investigation of the synergism of colistin and rifampin against MDR A. baumannii isolates via systematic review and meta-analysis. A systematic literature search was performed using the following 4 databases (PubMed, Scopus, EMBASE and ISI Web of Sciences). The related articles were evaluated during the period from December 2014 to January 2015. Information based on resistance and sensitivity to antibiotics, the minimum inhibitory concentration and the effects of two antibiotics on each other including synergism, antagonism, relative synergism and additive antagonism were extracted. A meta-analysis of 17 studies including 448 samples was brought into process and 2% (95% CI 0-4%) and 72% (95% CI 56-89%) resistance to colistin and rifampin were observed, respectively. 42% of all isolates showed MIC = 4 µg/ml (95% CI 14-69%) to rifampin and 30% MIC= 2 µg/ml to colistin (95% CI 3.8-78%). MIC50 and MIC90 for both rifampin and colistin were 2 µg/ml and 4 µg/ml, respectively. 63% of the strains demonstrated synergy (95% CI 37-90%), 7% were highlighted as relative synergism (95% CI 0.0- 13%), 3% showed an additive effect (95% CI -0.0-7%) and 14% were indifferent (95% CI 6-23%). The antagonistic effect was not observed in this combination. Synergy rates of time-kill assay in rifampin and colistin combinations were generally higher than those of check bored microdilution and E-test method. The results demonstrated that the combination therapy could be more useful when compared to monotherapy and that this strategy might reduce the resistance rate to rifampin in MDR A. baumannii isolates.


Patient and Provider Perspectives on a Decision Aid for Familial Hypercholesterolemia.
Familial Hypercholesterolemia (FH) is an inherited disorder associated with increased cardiovascular risk that requires patients to make multiple impactful decisions regarding the management of their condition. Patient decision aids (PDAs) can facilitate shared decision-making (SDM) and enable patients to make choices that are concordant with their goals and values. To inform the development of a PDA for patients with FH, we employed a qualitative inductive approach and focus group discussions with patients, physicians, and genetic counselors. We explored and analyzed the perceptions and understanding of these stakeholders related to a PDA for FH and identified important concepts to include in the PDA. Categories emerging from focus group discussions included: (a) perceptions of a PDA related to FH; (b) discussion about the content of a PDA related to FH; and (c) SDM. In general, stakeholders were in favor of developing tools which can inform and individualize discussion about genetic testing and treatment options for FH. Physicians valued a tool that facilitates knowledge transfer to FH patients. Patients desired a tool to help them understand the genetic aspects of and treatment options related to FH. Genetic counselors emphasized the inclusion of visual aids to support discussion with patients. Potential barriers to and facilitators of PDA implementation were identified. The input of various stakeholders will inform the development of a prototype tool that will be iteratively tested before implementation in the clinical setting.


Structural and catalytic properties of CMP kinase from Bacillus subtilis: a comparative analysis with the homologous enzyme from Escherichia coli.
CMP kinases from Bacillus subtilis and from Escherichia coli are encoded by the cmk gene (formerly known as jofC in B. subtilis and as mssA in E. coli). Similar in their primary structure (43% identity and 67% similarity in amino acid sequence), the two proteins exhibit significant differences in nucleotide binding and catalysis. ATP, dATP, and GTP are equally effective as phosphate donors with E. coli CMP kinase whereas GTP is a poor substrate with B. subtilis CMP kinase. While CMP and dCMP are the best phosphate acceptors of both CMP kinases, the specific activity with these substrates and ATP as donor are 7- to 10-fold higher in the E. coli enzyme; the relative Vm values with UMP and CMP are 0.1 for the B. subtilis CMP kinase and 0.01 for the E. coli enzyme. CMP increased the affinity of E. coli CMP kinase for ATP or for the fluorescent analog 3'-anthraniloyl dATP by one order of magnitude but had no effect on the B. subtilis enzyme. The differences in the catalytic properties of B. subtilis and E. coli CMP kinases might be reflected in the structure of the two proteins as inferred from infrared spectroscopy. Whereas the spectrum of B. subtilis CMP kinase is dominated by a band at 1633 cm-1 (representing beta type structures), the spectrum of the E. coli enzyme is dominated by two bands at 1653 and 1642 cm-1 associated with alpha-helical and unordered structures, respectively. CMP induced similar spectral changes in both proteins with a rearrangement of some of the beta-structures. ATP increases the denaturation temperature of B. subtilis CMP kinase by 9.3 degrees C, whereas in the case of the E. coli enzyme, binding of ATP has only a minor effect.


Effect of central nervous system metastases on treatment discontinuation and survival in older women receiving trastuzumab for metastatic breast cancer.
Background. Trastuzumab improves survival in HER2-positive women with metastatic breast cancer (MBC). The consequences of longer survival include a higher likelihood of additional metastases, including those in the central nervous system (CNS). The effect of CNS metastases on both trastuzumab discontinuation and survival in older patients has not been described. Patients and Methods. We used the Surveillance Epidemiology and End Results (SEER) Medicare data to identify a cohort of 562 women age 66 or older with MBC who were diagnosed between January 1, 2000 and December 31, 2005, free of CNS metastases, and initiated trastuzumab after MBC diagnosis. Time to discontinuation and time to death were analyzed using proportional hazards models. Results. Newly diagnosed CNS metastases were associated with both higher risk of trastuzumab discontinuation (relative hazard [RH] = 1.78, 95% CI 1.11-2.87) and higher risk of death (RH = 2.49, 95% CI 1.84-3.37). The incidence rate of new CNS metastases was comparable among various sites of metastasis (10.7 to 14.7 per 1,000 patient-months), except for bone which was higher (24.1 per 1,000). Conclusion. The diagnosis of CNS metastases was associated with an increase in both the likelihood of discontinuing trastuzumab therapy as well as the risk of death.


Long-term complementation of DNA repair deficient human primary fibroblasts by retroviral transduction of the XPD gene.
Due to their limited life time in culture and their relative resistance to DNA transfection, primary fibroblasts derived from UV-hypersensitive patients could not be used for cloning DNA repair gene and studying stable complementation with wild-type DNA repair genes. Primary cells were only used for complementation analysis after transient expression through cell fusion. DNA microinjection and transfection. We report the retroviral-mediated highly efficient transfer and stable expression of XPD/ERCC2 gene in fibroblast strains from eight different patients using the LXPDSN retroviral vector. Cells derived from skin biopsies of xeroderma pigmentosum and trichothiodystrophy patients were incubated with vector-containing suspension and selected with the neomycin-analog G418. LXPDSN vector specifically complemented cells belonging to the XP-D group. Long-term reversion of repair-deficient phenotype, monitored by UV survival and UDS analysis, has been achieved in these diploid fibroblasts. We demonstrate this methodology is a powerful tool to study phenotypic reversion of nucleotide excision repair-deficient cells such as cellular DNA repair properties and we suggest that it may be used to study other cellular parameters (cell cycle regulation, p53 stability or immunosurveillance-controlling factors) involved in UV-induced skin cancers and which reliability requires the use of untransformed cells.


The nonsteady state modeling of freeze drying: in-process product temperature and moisture content mapping and pharmaceutical product quality applications.
Theoretical models of the freeze-drying process are potentially useful to guide the design of a freeze-drying process as well as to obtain information not readily accessible by direct experimentation, such as moisture distribution and glass transition temperature, Tg, within a vial during processing. Previous models were either restricted to the steady state and/or to one-dimensional problems. While such models are useful, the restrictions seriously limit applications of the theory. An earlier work from these laboratories presented a nonsteady state, two-dimensional model (which becomes a three-dimensional model with an axis of symmetry) of sublimation and desorption that is quite versatile and allows the user to investigate a wide variety of heat and mass transfer problems in both primary and secondary drying. The earlier treatment focused on the mathematical details of the finite element formulation of the problem and on validation of the calculations. The objective of the current study is to provide the physical rational for the choice of boundary conditions, to validate the model by comparison of calculated results with experimental data, and to discuss several representative pharmaceutical applications. To validate the model and evaluate its utility in studying distribution of moisture and glass transition temperature in a representative product, calculations for a sucrose-based formulation were performed, and selected results were compared with experimental data. THEORETICAL MODEL: The model is based on a set of coupled differential equations resulting from constraints imposed by conservation of energy and mass, where numerical results are obtained using finite element analysis. Use of the model proceeds via a "modular software package" supported by Technalysis Inc. (Passage/ Freeze Drying). This package allows the user to define the problem by inputing shelf temperature, chamber pressure, container properties, product properties, and numerical analysis parameters required for the finite element analysis. Most input data are either available in the literature or may be easily estimated. Product resistance to water vapor flow, mass transfer coefficients describing secondary drying, and container heat transfer coefficients must normally be measured. Each element (i.e., each small subsystem of the product) may be assigned different values of product resistance to accurately describe the nonlinear resistance behavior often shown by real products. During primary drying, the chamber pressure and shelf temperature may be varied in steps. During secondary drying, the change in gas composition from pure water to mostly inert gas is calculated by the model from the instantaneous water vapor flux and the input pumping capacity of the freeze dryer. Comparison of the theoretical results with the experiment data for a 3% sucrose formulation is generally satisfactory. Primary drying times agree within two hours, and the product temperature vs. time curves in primary drying agree within about +/-1 degrees C. The residual moisture vs. time curve is predicted by the theory within the likely experimental error, and the lack of large variation in moisture within the vial (i.e., top vs. side vs. bottom) is also correctly predicted by theory. The theoretical calculations also provide the time variation of "Tg-T" during both primary and secondary drying, where T is product temperature and Tg is the glass transition temperature of the product phase. The calculations demonstrate that with a secondary drying protocol using a rapid ramp of shelf temperature, the product temperature does rise above Tg during early secondary drying, perhaps being a factor in the phenomenon known as "cake shrinkage." The theoretical results of in-process product temperature, primary drying time, and moisture content mapping and history are consistent with the experimental results, suggesting the theoretical model should be useful in process development and "trouble-shooting" applications.


Raman technologies in cancer diagnostics.
Despite significant effort, cancer still remains a leading cause of death worldwide. In order to reduce its burden, the development and improvement of noninvasive strategies for early detection and diagnosis of cancer are urgently needed. Raman spectroscopy, an optical technique that relies on inelastic light scattering arising from molecular vibrations, is one such strategy, as it can noninvasively probe cancerous markers using only endogenous contrast. In this review, spontaneous, coherent and surface enhanced Raman spectroscopies and imaging, as well as the fundamental principles governing the successful use of these techniques, are discussed. Methods for spectral data analysis are also highlighted. Utilization of the discussed Raman techniques for the detection and diagnosis of cancer in vitro, ex vivo and in vivo is described. The review concludes with a discussion of the future directions of Raman technologies, with particular emphasis on their clinical translation.


Comparable clinical efficacy and tolerability of 20 mg pantoprazole and 20 mg omeprazole in patients with grade I reflux oesophagitis.
Several clinical trials have shown that pantoprazole (40 mg) and omeprazole (40 or 20 mg) have similar efficacy and safety in the treatment of grade II-IV reflux oesophagitis (Savary-Miller classification). To compare the efficacy and safety of once-daily doses of pantoprazole (20 mg) and omeprazole (20 mg) with respect to symptom relief and healing of patients with grade I reflux oesophagitis. Patients with endoscopically established grade I reflux oesophagitis (non-confluent, patchy red lesions with/without white fibrin coating) were enrolled into this randomized, open, parallel-group, multicentre study. A total of 328 patients (n=166 in the pantoprazole group, n=162 in the omeprazole group) were recruited in 23 centres. Patients received 4 weeks of treatment. If the reflux oesophagitis was not completely healed, the treatment was extended to 8 weeks. After 2 and 4 weeks of treatment with either pantoprazole or omeprazole, the rate of symptom relief was similar (70% vs. 79% and 77% vs. 84%, respectively). High healing rates were observed after 4 and 8 weeks (pantoprazole: 84% and 90%, respectively; omeprazole: 89% and 95%, respectively). Both treatments were well tolerated. The most frequently reported adverse events on pantoprazole and omeprazole, respectively, were nausea (8% vs. 7%), diarrhoea (5% vs. 6%) and headache (6% vs. 3%). After 4 and 8 weeks of treatment with pantoprazole (20 mg) or omeprazole (20 mg), patients with mild gastro-oesophageal reflux disease (grade I) showed comparably high rates of symptom relief and healing. Both treatments were safe and well tolerated.


Enhanced activation of matrix metalloproteinase-9 correlates with the degree of papillary thyroid carcinoma infiltration.
To determine whether matrix metalloproteinase-9 (MMP-9) may be a useful adjunctive tool for predicting unfavorable biological behavior of papillary thyroid carcinoma (PTC) by evaluating the expression profile and proteolytic activity of MMP-9 in PTC by different techniques and correlating the findings with clinicopathological prognostic factors. Immunohistochemical localization of MMP-9 was analyzed with antibodies specific for either total or active MMP-9. Activation ratios of MMP-9 were calculated by quantifying gel zymography bands. Enzymatic activity of MMP-9 was localized by in situ zymography after inhibiting MMP-2 activity. Immunostaining of total and active MMP-9 was observed in tumor tissue and occasionally in non-neoplastic epithelium. Only active MMP-9 was significantly associated with extrathyroid invasion, lymph-node metastasis, and the degree of tumor infiltration (P<0.001, P=0.004, and P<0.001, respectively). Gelatin zymography revealed a correlation between the MMP-9 activation ratio and nodal involvement, extrathyroid invasion, and the degree of tumor infiltration. In situ zymography showed that gelatinases exerted their activity in tumor parenchymal and stromal cells. Moreover, after application of MMP-2 inhibitor, the remaining gelatinase activity, corresponding to MMP-9, was highest in cancers with the most advanced degree of tumor infiltration. This is the first report suggesting that the evaluation of active MMP-9 by immunohistochemistry and determination of its activation ratio by gelatin zymography may be a useful adjunct to the known clinicopathological factors in predicting tumor behavior. Most important, in situ zimography with an MMP-2 inhibitor for the first time demonstrated a strong impact of MMP-9 activity on the degree of tumor infiltration during PTC progression.


Applying n-dimensional hypervolumes for species delimitation: unexpected molecular, morphological, and ecological diversity in the Leaf-Toed Gecko Phyllodactylus reissii Peters, 1862 (Squamata: Phyllodactylidae) from northern Peru.
An integrative taxonomic approach based on morphology, molecular analyses, and climatic niche modeling was used to uncover cryptic diversity in the phyllodactylid gecko species Phyllodactylus reissii. At least three distinct species could be identified among the examined specimens from southern Ecuador and northern Peru. Phyllodactylus magister, described by Noble (1924) from arid Andean valleys of the Chinchipe and Marañón rivers in the Peruvian Department of Cajamarca and synonymized with P. reissii by Dixon & Huey (1970) is elevated from synonymy and a detailed redescription is provided. A new species of the genus Phyllodactylus from the Andean dry forest of the southern Marañón valley is identified and described herein. Phyllodactylus pachamama sp. nov. is differentiated from other South American congeners on the basis of mtDNA sequence divergence, morphological characters, and differences in the realized climatic niche. At least in Peru, P. reissii seems to primarily inhabit the northern coastal region west of the Andes, while the inter-Andean area along the Río Marañón and its tributaries seems to be inhabited mostly by other species of the genus, which are endemic to this area. The Andean valleys are underestimated in terms of biodiversity and lack thorough investigation and conservation actions.


Restoration and maintenance of intravenous insulin infusion in the vapor-pressure powered implantable infusion device.
Of the reasons to discontinue continuous implantable insulin pump therapy, flow rate reduction is the most common, occurring in 27 of 42 pumps in the University of Minnesota series. Thrombosis at the catheter tip appears to be a major reason for flow reduction. Six different procedures to restore flow in pumps were performed. Two of the procedures involve the infusion of an alkaline solution through the device, replacing the insulin/glycerol solution normally infused; the other four procedures are surgical ones involving manipulation of the catheter. In restoring flow the non-operative procedures achieved a success rate of 50%, having taken as long as 3 months to restore flow, and having allowed up to 1 year of further insulin pump therapy; flow in all the pumps so treated eventually decreased again. The operative procedures were nearly 100% successful, restored flow immediately, and allowed longer periods of adequate flow, but flow rate did again decrease. When flow rate reduction occurs further flow improvement procedures may be attempted without difficulty and are well tolerated. In implantable pumps the biocompatibility of the blood-catheter tip interface needs to be improved to deal with the recurrent problems of insulin infusion device flow rate decrease.


Cervical radiofrequency neurotomy reduces psychological features in individuals with chronic whiplash symptoms.
Individuals with chronic whiplash associated disorder (WAD) demonstrate various psychological features. It has previously been demonstrated that cervical radiofrequency neurotomy (cRFN) resolves psychological distress and anxiety. It is unknown if cRFN also improves or reduces a broader spectrum of psychological substrates now commonly identified in chronic whiplash, such as post-traumatic stress disorder (PTSD) and pain catastrophizing. To determine if reducing pain in the cervical spine (following cRFN) significantly reduces psychological features (distress, pain catastrophizing and post-traumatic stress symptoms) in individuals with chronic WAD. Tertiary spinal intervention centre in Calgary, Alberta, Canada. Prospective observational study of consecutive patients. Fifty-three individuals with chronic whiplash associated disorder symptoms (Grade 2). Cervical RFN following successful response to cervical facet joint blockade. Measures were made at 4 time points: 2 prior to RFN, and 1-month and 3-months post-RFN. Psychological measures included the General Health Questionnaire (GHQ-28); Pain Catastrophizing Scale (PCS) and the Post Traumatic Stress Diagnostic Scale (PDS). Self-reported pain (VAS) and disability (NDI) measures were also collected. Pain, disability, psychological distress and pain catastrophization significantly decreased at both 1-month and 3 months following cervical RFN. There was no significant change in post-traumatic stress symptom severity (P = 0.39). Reducing pain via cRFN was associated with significant improvement in psychological distress and pain catastrophizing, but not posttraumatic stress symptoms. Individual administering questionnaires was not blinded to aim(s) of the study. Other psychological features possibly present in WAD were not measured. Effective pain relief would seem a crucial element in the management of psychological features associated with chronic WAD. IRB Approval: University of Calgary Conjoint Health Research Ethics Board ID#: E-22082.


Impact of longitudinal power budget in coherent transmission systems employing digital back-propagation.
We report the impact of longitudinal signal power profile on the transmission performance of coherently-detected 112 Gb/s m-ary polarization multiplexed quadrature amplitude modulation system after compensation of deterministic nonlinear fibre impairments. Performance improvements up to 0.6 dB (Q(eff)) are reported for a non-uniform transmission link power profile. Further investigation reveals that the evolution of the transmission performance with power profile management is fully consistent with the parametric amplification of the amplified spontaneous emission by the signal through four-wave mixing. In particular, for a non-dispersion managed system, a single-step increment of 4 dB in the amplifier gain, with respect to a uniform gain profile, at ~2/3(rd) of the total reach considerably improves the transmission performance for all the formats studied. In contrary a negative-step profile, emulating a failure (gain decrease or loss increase), significantly degrades the bit-error rate.


Medical revalidation as professional regulatory reform: Challenging the power of enforceable trust in the United Kingdom.
For more than two decades, international healthcare crises and ensuing political debates have led to increasing professional governance and regulatory policy reform. Governance and policy reforms, commonly representing a shift from embodied trust in professionals to state enforceable trust, have challenged professional power and self-regulatory privileges. However, controversy remains as to whether such policies do actually shift the balance of power and what the resulting effects of policy introduction would be. This paper explores the roll-out and operationalisation of revalidation as medical regulatory reform within a United Kingdom National Health Service hospital from 2012 to 2013, and its impact upon professional power. Revalidation policy was subject to the existing governance and management structures of the organisation, resulting in the formal policy process being shaped at the local level. This paper explores how the disorganised nature of the organisation hindered rather than facilitated robust processes of professional governance and regulation, fostering formalistic rather than genuine professional engagement with the policy process. Formalistic engagement seemingly assisted the medical profession in retaining self-regulatory privileges whilst maintaining professional power over the policy process. The paper concludes by challenging the concept of state enforceable trust and the theorisation that professional groups are effectively regulated and controlled by means of national and organisational objectives, such as revalidation.


New global guidelines on sedentary behaviour and health for adults: broadening the behavioural targets.
In 2018, the World Health Organisation (WHO) commenced a program of work to update the 2010 Global Recommendations on Physical Activity for Health, for the first-time providing population-based guidelines on sedentary behaviour. This paper briefly summarizes and highlights the scientific evidence behind the new sedentary behaviour guidelines for all adults and discusses its strengths and limitations, including evidence gaps/research needs and potential implications for public health practice. An overview of the scope and methods used to update the evidence is provided, along with quality assessment and grading methods for the eligible new systematic reviews. The literature search update was conducted for WHO by an external team and reviewers used the AMSTAR 2 (Assessment of Multiple Systematic Reviews) tool for critical appraisal of the systematic reviews under consideration for inclusion. The Grading of Recommendations Assessment, Development and Evaluation (GRADE) method was used to rate the certainty (i.e. very low to high) of the evidence. The updated systematic review identified 22 new reviews published from 2017 up to August 2019, 14 of which were incorporated into the final evidence profiles. Overall, there was moderate certainty evidence that higher amounts of sedentary behaviour increase the risk for all-cause, cardiovascular disease (CVD) and cancer mortality, as well as incidence of CVD, cancer, and type 2 diabetes. However, evidence was deemed insufficient at present to set quantified (time-based) recommendations for sedentary time. Moderate certainty evidence also showed that associations between sedentary behaviour and all-cause, CVD and cancer mortality vary by level of moderate-to-vigorous physical activity (MVPA), which underpinned additional guidance around MVPA in the context of high sedentary time. Finally, there was insufficient or low-certainty systematic review evidence on the type or domain of sedentary behaviour, or the frequency and/or duration of bouts or breaks in sedentary behaviour, to make specific recommendations for the health outcomes examined. The WHO 2020 guidelines are based on the latest evidence on sedentary behaviour and health, along with interactions between sedentary behaviour and MVPA, and support implementing public health programmes and policies aimed at increasing MVPA and limiting sedentary behaviour. Important evidence gaps and research opportunities are identified.


Heterogeneity of depression. Classification of depressive subtypes by longitudinal course.
This paper describes the application of prospective longitudinal data from an epidemiological sample of young adults to define subtypes of major depression. Depression was classified on a spectrum from subthreshold manifestation of symptoms and duration at one end, to cases with recurrent episodes of depression meeting duration criteria for major depressive episodes at the other. There was a direct relationship between the severity of depression over the longitudinal course and both duration and recurrence of depressive episodes. The subgroup of depression with recurrence of both brief and longer duration episodes could be discriminated on most of the indicators of validity including symptoms, impairment, family history, and suicide attempts. In light of the young age of this cohort, the strong history of suicide attempts and other complications of depression among the subjects with recurrent depression was striking. These findings underscore the importance of employing course as a classification criterion of depression, and the inclusion of subthreshold episodes of depression in the characterization of course.


A Pharmacist Telephone Intervention to Identify Adherence Barriers and Improve Adherence Among Nonadherent Patients with Comorbid Hypertension and Diabetes in a Medicare Advantage Plan.
Patients with comorbid hypertension (HTN) and diabetes mellitus (DM) are at a high risk of developing macrovascular and microvascular complications of DM. Controlling high blood pressure can greatly reduce these complications. Angiotensin-converting enzyme inhibitors (ACEIs) or angiotensin II receptor blockers (ARBs) are recommended for patients with both DM and HTN by the American Diabetes Association guidelines, and their benefit and efficacy in reducing macrovascular and microvascular complications of DM have been well documented. Poor adherence, however, remains a significant barrier to achieving full effectiveness and optimal outcomes. To examine the effect of a brief pharmacist telephone intervention in identifying adherence barriers and improving adherence to ACEI/ARB medications among nonadherent patients with comorbid HTN and DM who are enrolled in a Medicare Advantage plan. Cigna-HealthSpring's medical claims data was used to identify patients with HTN and DM diagnoses by using ICD-9-CM codes 401 and 250, and at least 2 fills for ACEIs or ARBs between January 2013 and October 2013. Patients who failed to refill their medication for more than 1 day and had a proportion of days covered (PDC) < 0.8 were considered nonadherent and were contacted by a pharmacist by phone to identify adherence barriers. Two outcome variables were evaluated: The first was adherence to ACEIs/ARBs, defined as PDC during the 6 months following the phone call intervention. The second outcome variable was a categorical outcome of discontinuation versus continuation. Discontinuation was defined as not using ACEIs/ARBs during the 6-month post-intervention period. Patients who disenrolled from the plan in 2014 or were switched to another medication commonly used for treating DM and HTN were excluded from further analysis. Descriptive statistics were conducted to assess the frequency distribution of sample demographic characteristics at baseline. Multiple linear regression was conducted to assess the intervention effect on adherence during the 6 months post-intervention using the first outcome of post-intervention PDC, adjusting for baseline PDC and other covariates. Logistic regression was performed to assess the association between medication discontinuation and other baseline characteristics using the second outcome of discontinuation. Other control variables in the models included demographics (age, sex, language), physician specialty (primary care vs. specialist), health plan (low-income subsidy vs. other), Centers for Medicare & Medicaid risk score, Charlson Comorbidity Index, and number of distinct medications. In total, 186 hypertensive diabetic patients, nonadherent to ACEIs/ARBs (PDC < 0.8), were included in the study. Of the 186 patients, 87 received the pharmacist phone call intervention. Among these patients, forgetfulness (25.29%) and doctor issues, such as having difficulty scheduling appointments (16.79%), were the most commonly reported barriers. After excluding those who switched from ACEIs/ARBs to another medication, 157 patients were included in the logistic regression model. Of those, 131 had continued using ACEIs/ARBs and were included in the linear regression model. The mean (±SD) post-intervention PDC for the intervention group was 0.58 (±0.26) and for the control group 0.29 (±0.17). Intervention was a significant predictor of better adherence in the linear regression model after adjusting all the other baseline covariates (β = 0.3182, 95% CI = 0.19-0.38, P < 0.001). Other covariates were not significantly associated with better adherence. In the logistic regression model (discontinuation: 26 [yes]/131 [no]) for predicting medication discontinuation, patients who received intervention were more likely to continue using ACEIs/ARBs (OR = 3.56, 95% CI = 1.06-11.86), and those with a higher comorbidity index were less likely to continue using them (OR = 0.72, 95% CI = 0.53-0.99). The brief pharmacist telephone intervention resulted in significantly better PDCs during the 6 months following the intervention as well as lower discontinuation rates among a group of nonadherent patients with comorbid HTN and DM. The overall PDC rates in both the intervention and control groups were still lower than the recommended 80%. Improving adherence to clinically meaningful values may require more than a brief pharmacist phone call. Incorporating motivational interviewing techniques with follow-up calls to address adherence barriers may be more influential in forming sustainable behavioral change and enhancing medication adherence.


Synthetic Studies Toward the Skyllamycins: Total Synthesis and Generation of Simplified Analogues.
Herein, we report our synthetic studies toward the skyllamycins, a highly modified class of nonribosomal peptide natural products which contain a number of interesting structural features, including the extremely rare α-OH-glycine residue. Before embarking on the synthesis of the natural products, we prepared four structurally simpler analogues. Access to both the analogues and the natural products first required the synthesis of a number of nonproteinogenic amino acids, including three β-OH amino acids that were accessed from the convenient chiral precursor Garner's aldehyde. Following the preparation of the suitably protected nonproteinogenic amino acids, the skyllamycin analogues were assembled using a solid-phase synthetic route followed by a final stage solution-phase cyclization reaction. To access the natural products (skyllamycins A-C) the synthetic route used for the analogues was modified. Specifically, linear peptide precursors containing a C-terminal amide were synthesized via solid-phase peptide synthesis. After cleavage from the resin the N-terminal serine residue was oxidatively cleaved to a glyoxyamide moiety. The target natural products, skyllamycins A-C, were successfully prepared via a final step cyclization with concomitant formation of the unusual α-OH-glycine residue. Purification and spectroscopic comparison to the authentic isolated material confirmed the identity of the synthetic natural products.


Renal sympathetic responses to conflicting baroreceptor inputs: rapid ventricular pacing in dogs.
1. Ventricular tachycardia generates complex changes in baroreceptor input to the central nervous system: arterial baroreceptors are unloaded while cardiopulmonary receptors are stimulated. In humans with heart diseases, muscle sympathetic nerve activity increases during ventricular tachycardia. This suggests that arterial baroreceptor-mediated sympathoexcitation overrides cardiopulmonary receptor-mediated sympathoinhibition. However, the relative roles of each reflex are difficult to determine in humans. 2. We measured efferent renal sympathetic neural responses to simulated ventricular tachycardia, to determine what pathophysiological mechanisms are invoked when inputs from different baroreceptive areas change in opposite directions. In alpha-chloralose anaesthetized, mechanically ventilated dogs, we recorded the electrocardiogram, mean left atrial and arterial pressures and multifibre efferent renal sympathetic nerve activity (RSNA) during 1 min of right ventricular pacing at 214 beats min-1. Pacing was repeated after either sinoaortic or vagal cardiopulmonary denervation and again after both sinoaortic and cardiopulmonary denervation. 3. With all afferent baroreceptor pathways intact, right ventricular pacing elicited transient sympathoinhibition (delta RSNA, -19 +/- 10%, mean +/- S.E.M.). After sinoaortic denervation (cardiopulmonary receptors intact), right ventricular pacing elicited abrupt and sustained sympathoinhibition (delta RSNA, -53 +/- 8%, P < 0.05 vs. intact). After vagal cardiopulmonary denervation (sinoaortic receptors intact), right ventricular pacing elicited abrupt and sustained sympathoexcitation (delta RSNA, + 56 +/- 19%, P < 0.05 vs. intact). After both sinoaortic and vagal cardiopulmonary denervation, right ventricular pacing elicited a gradual increase in sympathetic outflow (delta RSNA, + 16 +/- 6%, P < 0.05 vs. intact). 4. We conclude that interactions between vagal cardiopulmonary and arterial baroreflexes determine renal sympathetic outflow during simulated ventricular tachycardia. In healthy anaesthetized dogs, the balance of the two opposing reflexes is weighted towards vagal cardiopulmonary-mediated sympathoinhibition.


Treatment of Tourette syndrome with delta-9-tetrahydrocannabinol (delta 9-THC): no influence on neuropsychological performance.
Previous studies provide evidence that marijuana (Cannabis sativa) and delta-9-tetrahydrocannabinol (Delta(9)-THC), the major psychoactive ingredient of marijuana, respectively, are effective in the treatment of tics and behavioral problems in Tourette syndrome (TS). It, therefore, has been speculated that the central cannabinoid receptor system might be involved in TS pathology. However, in healthy marijuana users there is an ongoing debate as to whether the use of cannabis causes acute and/or long-term cognitive deficits. In this randomized double-blind placebo-controlled study, we investigated the effect of a treatment with up to 10 mg Delta(9)-THC over a 6-week period on neuropsychological performance in 24 patients suffering from TS. During medication and immediately as well as 5-6 weeks after withdrawal of Delta(9)-THC treatment, no detrimental effect was seen on learning curve, interference, recall and recognition of word lists, immediate visual memory span, and divided attention. Measuring immediate verbal memory span, we even found a trend towards a significant improvement during and after treatment. Results from this study corroborate previous data suggesting that in patients suffering from TS, treatment with Delta(9)-THC causes neither acute nor long-term cognitive deficits. Larger and longer-duration controlled studies are recommended to provide more information on the adverse effect profile of THC in patients suffering from TS.


Assessment of genetic susceptibility to ethanol intoxication in mice.
Increased use of gene manipulation in mice (e.g., targeted or random mutagenesis) has been accompanied by increased reliance on a very few rapid and simple behavioral assays, each of which aspires to model a human behavioral domain. Yet, each assay comprises multiple traits, influenced by multiple genetic factors. Motor incoordination (ataxia), a common characteristic of many neurological disorders, may reflect disordered balance, muscle strength, proprioception, and/or patterned gait. Impaired motor performance can confound interpretation of behavioral assays of learning and memory, exploration, motivation, and sensory competence. The rotarod is one of the most commonly used tests to measure coordination in mice. We show here that exactly how the rotarod test is performed can markedly alter the apparent patterns of genetic influence both in undrugged performance and sensitivity to ethanol intoxication. However, when tested with well chosen parameters, the accelerating rotarod test showed very high inter- and intralaboratory reliability. Depending on test conditions, ethanol can either disrupt or enhance performance in some strains. Genetic contribution to performance on the accelerating versus the fixed-speed rotarod assay can be completely dissociated under some test conditions, and multiple test parameters are needed to assess the range of genetic influence adequately.


Seasonal variability of forest sensitivity to heat and drought stresses: A synthesis based on carbon fluxes from North American forest ecosystems.
Climate extremes such as heat waves and droughts are projected to occur more frequently with increasing temperature and an intensified hydrological cycle. It is important to understand and quantify how forest carbon fluxes respond to heat and drought stress. In this study, we developed a series of daily indices of sensitivity to heat and drought stress as indicated by air temperature (Ta ) and evaporative fraction (EF). Using normalized daily carbon fluxes from the FLUXNET Network for 34 forest sites in North America, the seasonal pattern of sensitivities of net ecosystem productivity (NEP), gross ecosystem productivity (GEP) and ecosystem respiration (RE) in response to Ta and EF anomalies were compared for different forest types. The results showed that warm temperatures in spring had a positive effect on NEP in conifer forests but a negative impact in deciduous forests. GEP in conifer forests increased with higher temperature anomalies in spring but decreased in summer. The drought-induced decrease in NEP, which mostly occurred in the deciduous forests, was mostly driven by the reduction in GEP. In conifer forests, drought had a similar dampening effect on both GEP and RE, therefore leading to a neutral NEP response. The NEP sensitivity to Ta anomalies increased with increasing mean annual temperature. Drier sites were less sensitive to drought stress in summer. Natural forests with older stand age tended to be more resilient to the climate stresses compared to managed younger forests. The results of the Classification and Regression Tree analysis showed that seasons and ecosystem productivity were the most powerful variables in explaining the variation of forest sensitivity to heat and drought stress. Our results implied that the magnitude and direction of carbon flux changes in response to climate extremes are highly dependent on the seasonal dynamics of forests and the timing of the climate extremes.


Apoptosis in rheumatoid arthritis synovium.
RA synovial tissue (ST) was studied to determine if and where apoptosis occurs in situ. Genomic DNA was extracted from 5 RA and 1 osteoarthritis ST samples. Agarose gel electrophoresis demonstrated DNA ladders characteristic for apoptosis from each tissue. In situ and labeling (ISEL) was used to identify DNA strand breaks consistent with apoptosis in frozen sections. 12 RA and 4 osteoarthritis ST were studied by ISEL and all were positive, but only 2 of 4 normal tissues were positive. The primary location of apopotic cells was the synovial lining. Some sublining cells were also positive, but lymphoid aggregate staining was conspicuously absent. Immunohistochemistry and ISEL were combined and showed that the lining cells with DNA strand breaks were mainly macrophages, although some fibroblastlike cells were also labeled. Sublining cells with fragmented DNA included macrophages and fibroblasts, but T cells in lymphoid aggregates, which expressed large amounts of bcl-2, were spared. DNA strand breaks in cultured fibroblastlike synoviocytes was assessed using ISEL. Apoptosis could be induced by actinomycin D, anti-fas antibody, IL-1, and TNF-alpha but not by IFN-gamma. Fas expression was also detected on fibroblast-like synoviocytes using flow cytometry. Therefore, DNA strand breaks occur in synovium of patients with arthritis. Cytokines regulate this process, and the cytokine profile in RA (high IL-1/TNF; low IFN-gamma) along with local oxidant injury might favor induction of apoptosis.


Primary structure and microtubule-interacting domain of the SP-H antigen: a mitotic MAP located at the spindle pole and characterized as a homologous protein to NuMA.
Using a human autoantibody, SP-H, we identified a 200-230 kDa mitotic MAP in a variety of mammalian cell lines which shows affinity for the minus end of microtubules and also becomes associated with the spindle pole during mitosis. To examine the detailed structure and functional organization of the protein, the gene coding for the end-specific MAP was isolated and characterized by screening a human placenta lambda gt11 expression library using SP-H as a probe. Overlapping cDNA clones, which covered the entire length of the coding region of the SP-H antigen, were obtained. Polyclonal antibodies raised against fusion proteins generated from non-overlapping cDNA fragments stained the HeLa SP-H antigen in interphase and mitotic cells, and recognized a single 215 kDa band on immunoblots, as did the original SP-H antibody. Analysis of the nucleotide sequence revealed a 7,091 nucleotide sequence with an open reading frame of 6,345 nucleotides encoding a 2,115 amino acid polypeptide with a calculated molecular mass of 238,376 Da. The predicted amino acid sequence showed the protein to be composed of an alpha-helical domain, flanked by globular domains located at the amino and carboxy termini. The sequence contained five repeats of the hypothetical leucine zipper motif: one is in the N-terminal globular domain, and four are in the central alpha-helical stalk. Comparison with other sequences in the database shows that the SP-H antigen is identical to the NuMA protein reported by Yang et al. (1992) J. Cell Biol. 116, 1303-1317, but there are differences between the SP-H antigen and NuMA sequence reported by Compton et al. (1992) J. Cell Biol. 116, 1395-1408. cDNA inserts of the truncated SP-H antigen were expressed in both insect Sf9 cells and in cultured mammalian cells. The recombinant protein corresponding to the C-terminal half of the protein was restricted to the nucleus, whereas the N-terminal half of the protein was localized in the cytoplasm, suggesting the presence of a nuclear translocation signal(s) in the C-terminal domain. The C-terminal polypeptide expressed in mitotic COS cells was shown to specifically localize at the spindle pole. Microtubule-binding assays using in vitro transcribed/translated polypeptide products from different domains of the SP-H antigen further suggested that the SP-H antigen interacts with microtubules through the globular domain at the C-terminus.


Spatially incoherent common-path off-axis color digital holography.
We describe a new method for recording spatially incoherent common-path off-axis color digital holograms. We present the theoretical and experimental evidence to demonstrate an incoherent common-path off-axis color digital holographic (ICOCH) system capable of capturing information from three-dimensional color objects under incoherent illumination, both in transmission and reflection modes. Fresnel incoherent correlation holography (FINCH), a common-path system, is a frequently used incoherent holography technique. Our proposed system is conceptually similar to an advanced form of FINCH; moreover, it has three advantages over this advanced form of FINCH. First, removal of the spatial light modulator makes our system simpler and more cost-effective. Second, removal of the polarizer or analyzer allows for greater light throughput. Third, the off-axis optical configuration enables separation of zero-order and twin images with only a single exposure per color rather than requiring three exposures per color for in-line holography FINCH. Therefore, we believe that this simple and cost-effective system with high light throughput can acquire incoherent holograms for different colors involving single exposure for each color, which makes the ICOCH system suitable for many applications.


The impact of ERI, burnout, and caring for SARS patients on hospital nurses' self-reported compliance with infection control.
Siegrist's (1996) Effort-Reward Imbalance (ERI) Model provided the theoretical basis for this secondary data analysis that examines the relationship between nurses' ERI and their self-reported compliance with infection control, between ERI and burnout and nurses' compliance, and between nurses' experience in caring for SARS patients and their compliance with infection control. Data for this study came from a collaborative interdisciplinary study examining the barriers and facilitators to implementing protective measures against SARS and other existing and emerging infections among hospital nurses in Ontario and British Columbia. This is the first study to examine the relationship between ERI and compliance with infection control, as well as the impact of nurses' experience in caring for SARS patients on their compliance behaviour with infection control. Hierarchical multiple linear regression analyses revealed that ERI is a significant predictor of decreased compliance with infection control (beta = -.15, p < .05). While ERI was shown to be associated with burnout (beta = .60, p < .001), the combined effect of these two variables did not significantly improve the prediction of compliance behaviour (beta = -.03, p = .63). Nurses who reported having directly cared for SARS patients were found to have increased compliance with infection control (beta = .15, p < .001) after controlling for demographic and work environment factors. These findings highlight how nurses' adverse workplace environments can affect their work and health and thus, can be used by nursing and hospital administrators to help develop interventions to lower occupational stress and improve health in the workplace.


Switching to sanitation: Understanding latrine adoption in a representative panel of rural Indian households.
Open defecation, which is still practiced by about a billion people worldwide, is one of the most compelling examples of how place influences health in developing countries. Efforts by governments and development organizations to address the world's remaining open defecation would be greatly supported by a better understanding of why some people adopt latrines and others do not. We analyze the 2005 and 2012 rounds of the India Human Development Survey (IHDS), a nationally representative panel of households in India, the country which is home to 60% of the people worldwide who defecate in the open. Among rural households that defecated in the open in 2005, we investigate what baseline properties and what changes over time are associated with switching to latrine use between 2005 and 2012. We find that households that are richer or better educated, that have certain demographic properties, or that improved their homes over this period were more likely to switch to using a latrine or toilet. However, each of these effect sizes is small; overall switching to latrine use from open defecation is low; and no ready household-level mechanisms are available for sanitation programs to widely influence these factors. Our research adds to a growing consensus in the literature that the social context should not be overlooked when trying to understand and bring about change in sanitation behavior.


Repeated movie viewings produce similar local activity patterns but different network configurations.
People seek novelty in everyday life, but they also enjoy viewing the same movies or reading the same novels a second time. What changes and what stays the same when re-experiencing a narrative? In examining this question with functional neuroimaging, we found that brain activity reorganizes in a hybrid, scale-dependent manner when individuals processed the same audiovisual narrative a second time. At the most local level, sensory systems (occipital and temporal cortices) maintained a similar temporal activation profile during the two viewings. Nonetheless, functional connectivity between these same lateral temporal regions and other brain regions was stronger during the second viewing. Furthermore, at the level of whole-brain connectivity, we found a significant rearrangement of network partition structure: lateral temporal and inferior frontal regions clustered together during the first viewing but merged within a fronto-parietal cluster in the second. Our findings show that repetition maintains local activity profiles. However, at the same time, it is associated with multiple network-level connectivity changes on larger scales, with these changes strongly involving regions considered core to language processing.


Genetic relationships of serologically nontypable and serotype b strains of Haemophilus influenzae.
A collection of 242 strains of Haemophilus influenzae, including 65 nontypable (unencapsulated) isolates and 177 encapsulated serotype b isolates recovered largely from children with invasive and noninvasive diseases in the United States, was characterized by the electrophoretic mobilities of 15 metabolic enzymes presumably encoded by chromosomal genes. All enzymes were polymorphic for three to seven electromorphs, and 94 distinctive multilocus genotypes (electrophoretic types [ETs]) were distinguished, among which mean genetic (allelic) diversity was 0.500. Isolates recovered from cases of invasive or noninvasive diseases did not differ significantly in level of genetic variation. The observation that 29 ETs were represented exclusively by serotype b isolates and that each of the 65 nontypable isolates was of a unique ET strongly confirmed the hypothesis that unencapsulated clinical isolates are not merely phenotypic variants of the common serotype b cell lines. Rather, the two types of isolates are distinctive subsets of the multilocus chromosomal genotypes of the species as a whole. Serotype b capsule occurred in three groups of isolates that are distantly related in multilocus enzyme genotype. Isolates of four closely related nontypable biotype IV ETs associated with obstetrical infections or neonatal bacteremia were highly divergent from all others examined and may be specifically distinct. A phylogenetic scenario was proposed in which the ancestor of H. influenzae was encapsulated and the nontypable clones arose by convergent evolutionary loss of the ability to synthesize or extracellularly express a polysaccharide capsule.


European Code against Cancer 4th Edition: Alcohol drinking and cancer.
Alcohol consumption is the third leading risk factor for disease and mortality in Europe. The International Agency for Research on Cancer (IARC) Monographs provide strengthened evidence that the consumption of alcoholic beverages is causally associated with cancers of the oral cavity, pharynx, larynx, oesophagus, liver, colorectum and female breast, even for low and moderate alcohol intakes. The risk of cancer increases in a dose-dependent manner, and the higher the amount of alcohol consumed, the higher the risk of developing cancer. Several biological mechanisms explain the carcinogenicity of alcohol; among them, ethanol and its genotoxic metabolite acetaldehyde play a major role. Taking all this evidence into account, a recommendation of the 4th edition of the European Code against Cancer (ECAC) is: "If you drink alcohol of any type, limit your intake. Not drinking alcohol is better for cancer prevention."


Selective dispersal of avian rhombomere cells in orthotopic and heterotopic grafts.
During hindbrain development, cells become segregated into segmental groups, rhombomeres, by mechanisms that are presently unknown. One contributory factor early in development may be an alternating periodicity in cell surface properties down the neuraxis. This possibility was previously suggested by experiments in which tissue from different segmental levels was apposed in the absence of a boundary. New boundaries were regenerated only when rhombomeres from adjacent positions or positions three rhombomeres distant from one another were apposed. Combinations of two odd-numbered or two even-numbered rhombomeres usually failed to generate a boundary. In order to pursue this phenomenon to the cellular level, we have used two approaches, both involving donor-to-host transplantation. First, quail rhombomeres were grafted at various hindbrain levels of a chick host. Apposition of rhombomere 4 (r4) with r3 was concomitant with negligible cell mixing across the interface. By contrast, combinations of r3 with r5 or with r3 tissue led to cell mixing that was more extensive in combinations of identical rhombomeres (r3 with r3) than between two alternate ones (r3 with r5). Secondly, we grafted small pieces of fluorescently prelabelled chick rhombomere tissue at various hindbrain levels of chick hosts. In most cases, cells dispersed widely when transplanted orthopically or two segments distant from that of their origin. Cells transplanted into an adjacent segment, however, showed a tendency to remain undispersed. Among the different graft combinations, furthermore, there was a variation in the extent of dispersal that showed an additional level of complexity not revealed in boundary regeneration experiments. The possibility is raised that the early partitioning of rhombomeres involves a hierarchy in the adhesive preferences of cell-cell interactions along the neuraxis.


The influence of species interactions on geographic range change under climate change.
The fossil record tells us that many species shifted their geographic distributions during historic climate changes, but this record does not portray the complete picture of future range change in response to climate change. In particular, it does not provide information on how species interactions will affect range shifts. Therefore, we also need modern research to generate understanding of range change. This paper focuses on the role that species interactions play in promoting or preventing geographic ranges shifts under current and future climate change, and we illustrate key points using empirical case studies from an integrated study system. Case studies can have limited generalizability, but they are critical to defining possible outcomes under climate change. Our case studies emphasize host limitation that could reduce range shifts and enemy release that could facilitate range expansion. We also need improvements in modeling that explicitly consider species interactions, and this modeling can be informed by empirical research. Finally, we discuss how species interactions have implications for range management by people.


A novel fluorescent sensitive assay for detection of differential T cell mediated lysis of multiple adherent target cells.
There are few studies that have investigated T cell mediated lysis of adherent cells. We have developed a novel, rapid and sensitive fluorescent dye-swap assay that allows efficient detection of adherent target cell lysis. The assay allows simultaneous use of multiple differentially sensitised targets and facilitates concomitant surface or intracellular effector cell phenotypic analysis.


How the sea squirt nucleus tells mesoderm not to be endoderm.
Sea squirts are simple invertebrate chordates. In this issue of Developmental Cell, Takatori et al. show nuclear migration within ascidian mesendodermal cells enables polarized localization of Not mRNA, which encodes a homeobox protein that distinguishes mesoderm from endoderm fates. The link between nuclear migration and mRNA localization suggests exciting parallels with protostomes.


Fragile X and autism: Intertwined at the molecular level leading to targeted treatments.
Fragile X syndrome (FXS) is caused by an expanded CGG repeat (> 200 repeats) in the 5' untranslated portion of the fragile mental retardation 1 gene (FMR1), leading to deficiency or absence of the FMR1 protein (FMRP). FMRP is an RNA carrier protein that controls the translation of several other genes that regulate synaptic development and plasticity. Autism occurs in approximately 30% of FXS cases, and pervasive developmental disorder, not otherwise specified (PDD-NOS) occurs in an additional 30% of cases. Premutation repeat expansions (55 to 200 CGG repeats) may also give rise to autism spectrum disorders (ASD), including both autism and PDD-NOS, through a different molecular mechanism that involves a direct toxic effect of the expanded CGG repeat FMR1 mRNA. RNA toxicity can also lead to aging effects including tremor, ataxia and cognitive decline, termed fragile X-associated tremor ataxia syndrome (FXTAS), in premutation carriers in late life. In studies of mice bearing premutation expansions, there is evidence of early postnatal neuronal cell toxicity, presenting as reduced cell longevity, decreased dendritic arborization and altered synaptic morphology. There is also evidence of mitochondrial dysfunction in premutation carriers. Many of the problems with cellular dysregulation in both premutation and full mutation neurons also parallel the cellular abnormalities that have been documented in autism without fragile X mutations. Research regarding dysregulation of neurotransmitter systems in FXS, including the metabotropic glutamate receptor (mGluR)1/5 pathway and γ aminobutyric acid (GABA)A pathways, have led to new targeted treatments for FXS. Preliminary evidence suggests that these new targeted treatments will also be beneficial in non-fragile X forms of autism.


Topical tacrolimus for the treatment of psoriasis on the face, genitalia, intertriginous areas and corporal plaques.
Tacrolimus is an immunosuppressive drug that has proved effective in the treatment of psoriasis when administered systemically. Topically, it seems only useful in thin psoriasis plaques located on the face, genitalia, and intertriginous areas. We present an open-label clinical trial to test the efficacy of 0.1% tacrolimus ointment in patients with psoriasis on the face, intertriginous areas, both, and in corporal plaques. Efficacy was assessed with the evaluation of erythema, desquamation, infiltration, reduction of the PASI, and reduction of itching. A total of 15 patients were enrolled in the study. In all the localizations evaluated, each of the signs (erythema, desquamation, and infiltration) showed a statistically significant improvement when compared to the baseline (p < .001). Itching also improved rapidly. PASI was also reduced from a mean of 12 at baseline to 2.2 at the end of the study. Of the 15 patients, only 2 experienced an adverse effect (13%), which was described as a warm sensation in facial lesions which was transient and self-limited. In conclusion, tacrolimus ointment may be an alternative to classical options for the treatment of psoriasis, not only for intertriginous, genital, and facial areas, but also for corporal plaques without occlusion, with good tolerance.


Robustness to mechanical perturbations of center-launching technique for transparent board-to-board and data server interconnects.
Center-launching technique appears as a promising method to allow single-mode propagation in multi-mode fibers, guaranteeing full transparency to the transmitted optical signal also for applications in board-to-board and data server interconnects. In this paper we show that this technique is robust to mechanical perturbations up to about 1 kHz, demonstrating that the vibrations do not affect the transmission performances. Different experimental configurations are tested in order to exclude multimode propagation and to confirm the only fundamental mode propagation. Finally, a theoretical discussion comments the experimental results.


Increasing incidence of non-Hodgkin's lymphoma in Karachi, 1995-2002.
This first population-based study of non- Hodgkin lymphoma (NHL) from any region in Pakistan, provides an overview of the incidence pattern and time trends in Karachi and generates hypotheses for future experimental research. Epidemiological data for 429 incident (1(st) Jan 1995 to 31(st) Dec 2002), microscopically verified nodal and extra-nodal NHL cases, registered at the Karachi Cancer Registry (KCR) for Karachi South, were reviewed. The age standardized incidence rate (ASIR) was 5.3/100,000 in males (M) and 4.1/100,000 in females (F), in 1995. A gradual increase in the annual incidence was observed during the study period, with NHL incidence rate increasing in 2002 to 8.4/100,000 in men and 6.5/100,000 in women, almost double the 1995 rates. NHL affected all age groups in both genders and for each group the ASIR was higher among men than women, with an overall gender ratio of 1.9. The mean ages of the patients were 41.5 years (95% CI 39.1; 43.8) in males and 44.0 years (95% CI 40.8; 47.1) in females. The adult to childhood ratios were 8.6 (M) and 10.7 (F). B-cell NHL comprised 81.0% of NHL in males and 87.3% in females. One fourth of the NHL cases were extra-nodal, the largest group was of gastrointestinal origin (54.1% M, 38.5% F). The gastric component was 21% M and 25.6% F. Odds Ratios for sex, age-groups, ethnicity, religion, and subdivision by socio-economic categories were calculated by considering all malignancies, except lymphoproliferative disorders as controls. The odds ratio (OR) in men was 2.2 (95% CI 0.6; 3.0). Children and adolescents were at the highest risk of developing NHL, especially the 5-9 year olds, in both genders. A marginally higher risk was observed for the lower socioeconomic categories and for ethnicities belonging to Northern and North Western Pakistan (Punjabi, Pushtu and Baluch) residing in Karachi South. The incidence rates of NHL registered in Karachi South are likely to be a reflection of non-AIDS-associated NHL. Estimated HIV/AIDS incidence was too low during the study period in this population to have an impact on NHL incidence. The preponderance of low and intermediate grade lymphomas, paucity of central nervous system NHL and a higher childhood NHL component support this hypothesis. As yet unpublished reports, however, are raising the alarm on rising HIV positivity. NHL correlation with HIV/AIDS status and studies identifying risk factors of non- HIV/AIDS associated NHL (childhood viral infections, Hepatitis C virus, and Helicobacter pylori) are potential areas for future experimental and epidemiological research.


Allograft-prosthesis composite reconstruction of the proximal part of the humerus: functional outcome and survivorship.
Limb salvage following resection of a tumor in the proximal part of the humerus poses many challenges. Reconstructive options are limited because of the loss of periarticular soft-tissue stabilizers of the glenohumeral joint in addition to the loss of bone and articular cartilage. The purpose of this study was to evaluate the functional outcome and survival of the reconstruction following use of a humeral allograft-prosthesis composite for limb salvage. An allograft-prosthesis composite was used to reconstruct a proximal humeral defect following tumor resection in thirty-six consecutive patients at one institution over a sixteen-year period. The reconstruction was performed at the time of a primary tumor resection in thirty cases, after a failure of a reconstruction following a previous tumor resection in five patients, and following excision of a local recurrence in one patient. The mean duration of follow-up of the living patients was five years. Glenohumeral stability, function, implant survival, fracture rate, and union rate following the reconstructions were measured. Functional outcome and implant survival were analyzed on the basis of the amount of deltoid resection, whether the glenohumeral resection had been extra-articular or intra-articular, and the length of the humerus that had been resected. One patient sustained a glenohumeral dislocation. Deltoid resection (partial or complete) resulted in a reduced postoperative range of motion in flexion and abduction but had no effect on the mean Musculoskeletal Tumor Society score. Extra-articular resections were associated with lower Musculoskeletal Tumor Society scores. All patients had either mild or no pain and normal hand function at the time of final follow-up. The overall estimated rate of survival of the construct, with revision as the end point, was 88% at ten years. There were three failures due to progressive prosthetic loosening that necessitated removal of the construct. Four patients required an additional bone-grafting procedure to treat a delayed union of the osteosynthesis site. An allograft-prosthesis composite used for limb salvage following tumor resection in the proximal part of the humerus is a durable construct associated with an acceptable complication rate. Deltoid preservation and intra-articular resection are associated with a greater range of shoulder motion and a superior functional outcome, respectively.


Alfalfa dodder (Cuscuta campestris) toxicity in horses: clinical, haematological and serum biochemical findings.
The objective of this observational study is to describe clinical, haematological and serum biochemical findings of horses affected with alfalfa dodder (Cuscuta campestris) toxicity. Twenty horses naturally exposed to alfalfa dodder toxicity were examined and information was collected on history and clinical signs. Physical examination was done on horses in the premises (n=20), and venous blood samples of 12 horses were submitted for haematology and serum biochemical examination for each horse. Abnormal clinical signs started around 36 hours after horses were fed the contaminated alfalfa. Abnormal signs were seen in 11 horses and those included diarrhoea (n=8), decreased appetite (n=7), neurological signs (n=4) and abdominal pain (n=1). Some horses had multiple clinical signs of the above. The results of complete blood cell count revealed leukocytopenia, neutropenia and thrombocytopenia. Serum biochemical analysis revealed decreased ALP, AST and CPK levels and increased direct bilirubin level. The used alfalfa was stopped immediately and a different alfalfa from a new container that did not contain any weeds was fed. Horses on the premises were observed closely, and the abnormal clinical signs resolved within three days. No treatment was implemented. Knowledge about toxicity of horses by Cuscuta species is scarce in the English veterinary literature and very limited.


Recovery of locomotion after injury in Drosophila melanogaster depends on proprioception.
Locomotion is necessary for survival in most animal species. However, injuries to the appendages mediating locomotion are common. We assess the recovery of walking in Drosophila melanogaster following leg amputation. Whereas flies pre-amputation explore open arenas in a symmetric fashion on average, foreleg amputation induces a strong turning bias away from the side of the amputation. However, we find that unbiased walking behavior returns over time in wild-type flies, while recovery is significantly impaired in proprioceptive mutants. To identify the biomechanical basis of this locomotor impairment and recovery, we then examine individual leg motion (gait) at a fine scale. A minimal mathematical model that links neurodynamics to body mechanics during walking shows that redistributing leg forces between the right and left side enables the observed recovery. Altogether, our study suggests that proprioceptive input from the intact limbs plays a crucial role in the behavioral plasticity associated with locomotor recovery after injury.


Multiwavelength-integrated local model fitting method for interferometric surface profiling.
The local model fitting (LMF) method is a useful single-shot surface profiling algorithm that features fast measurement speed and robustness against vibration. However, the measurement range of the LMF method (i.e., measurable height difference between two neighboring pixels) is limited up to a quarter of the light source wavelength. To cope with this problem, the multiwavelength-matched LMF(MM-LMF) method was proposed, where the plain LMF method is first applied individually to interference images obtained from multiple light sources with different wavelengths, and then the LMF solutions are matched to obtain a range-extended solution. Although the MM-LMF method was shown to provide high measurement accuracy under moderate noise, phase unwrapping errors can occur if individual LMF solutions are erroneous. In this paper, we propose the multiwavelength-integrated LMF (MI-LMF) method, which directly computes a range-extended solution from multiple interference images in an integrated way. The effectiveness of the proposed MI-LMF method is demonstrated through simulations and actual experiments.


Delinquency, parental involvement, early adult criminality, and sex: Evidence of moderated mediation.
One purpose of this study was to determine whether parental involvement, measured in late adolescence, mediates the relationship between delinquency in mid-adolescence and crime in early adulthood. This study's second purpose was to ascertain whether this relationship is moderated by sex, such that late adolescent parental involvement mediates the delinquency-crime relationship in females but not in males. A secondary analysis of data provided by 579 (272 males, 307 females) members of the National Longitudinal Survey of Youth-Child (NLSYC) was conducted in an effort to evaluate the possibility of moderated mediation in the relationship between delinquency at age 16, parental involvement at age 18, and criminality at age 24. Moderated mediation analysis, path analysis, and causal mediation analysis revealed the presence of a conditional indirect relationship between delinquency, parental involvement, and adult crime moderated by sex. These results are consistent with views on cumulative disadvantage and gendered pathways to crime.


Social regulation of gene expression in human leukocytes.
Social environmental influences on human health are well established in the epidemiology literature, but their functional genomic mechanisms are unclear. The present study analyzed genome-wide transcriptional activity in people who chronically experienced high versus low levels of subjective social isolation (loneliness) to assess alterations in the activity of transcription control pathways that might contribute to increased adverse health outcomes in social isolates. DNA microarray analysis identified 209 genes that were differentially expressed in circulating leukocytes from 14 high- versus low-lonely individuals, including up-regulation of genes involved in immune activation, transcription control, and cell proliferation, and down-regulation of genes supporting mature B lymphocyte function and type I interferon response. Promoter-based bioinformatic analyses showed under-expression of genes bearing anti-inflammatory glucocorticoid response elements (GREs; p = 0.032) and over-expression of genes bearing response elements for pro-inflammatory NF-kappaB/Rel transcription factors (p = 0.011). This reciprocal shift in pro- and anti-inflammatory signaling was not attributable to differences in circulating cortisol levels, or to other demographic, psychological, or medical characteristics. Additional transcription control pathways showing differential activity in bioinformatic analyses included the CREB/ATF, JAK/STAT, IRF1, C/EBP, Oct, and GATA pathways. These data provide the first indication that human genome-wide transcriptional activity is altered in association with a social epidemiological risk factor. Impaired transcription of glucocorticoid response genes and increased activity of pro-inflammatory transcription control pathways provide a functional genomic explanation for elevated risk of inflammatory disease in individuals who experience chronically high levels of subjective social isolation.


Three-dimensional printed models for surgical planning of complex congenital heart defects: an international multicentre study.
To evaluate the impact of 3D printed models (3D models) on surgical planning in complex congenital heart disease (CHD). A prospective case-crossover study involving 10 international centres and 40 patients with complex CHD (median age 3 years, range 1 month-34 years) was conducted. Magnetic resonance imaging and computed tomography were used to acquire and segment the 3D cardiovascular anatomy. Models were fabricated by fused deposition modelling of polyurethane filament, and dimensions were compared with medical images. Decisions after the evaluation of routine clinical images were compared with those after inspection of the 3D model and intraoperative findings. Subjective satisfaction questionnaire was provided. 3D models accurately replicate anatomy with a mean bias of -0.27 ± 0.73 mm. Ninety-six percent of the surgeons agree or strongly agree that 3D models provided better understanding of CHD morphology and improved surgical planning. 3D models changed the surgical decision in 19 of the 40 cases. Consideration of a 3D model refined the planned biventricular repair, achieving an improved surgical correction in 8 cases. In 4 cases initially considered for conservative management or univentricular palliation, inspection of the 3D model enabled successful biventricular repair. 3D models are accurate replicas of the cardiovascular anatomy and improve the understanding of complex CHD. 3D models did not change the surgical decision in most of the cases (21 of 40 cases, 52.5% cases). However, in 19 of the 40 selected complex cases, 3D model helped redefining the surgical approach.


Glucagon regulates hepatic lipid metabolism via cAMP and Insig-2 signaling: implication for the pathogenesis of hypertriglyceridemia and hepatic steatosis.
Insulin induced gene-2 (Insig-2) is an ER-resident protein that inhibits the activation of sterol regulatory element-binding proteins (SREBPs). However, cellular factors that regulate Insig-2 expression have not yet been identified. Here we reported that cyclic AMP-responsive element-binding protein H (CREBH) positively regulates mRNA and protein expression of a liver specific isoform of Insig-2, Insig-2a, which in turn hinders SREBP-1c activation and inhibits hepatic de novo lipogenesis. CREBH binds to the evolutionally conserved CRE-BP binding elements located in the enhancer region of Insig-2a and upregulates its mRNA and protein expression. Metabolic hormone glucagon and nutritional fasting activated CREBH, which upregulated expression of Insig-2a in hepatocytes and inhibited SREBP-1c activation. In contrast, genetic depletion of CREBH decreased Insig-2a expression, leading to the activation of SREBP-1c and its downstream lipogenic target enzymes. Compromising CREBH-Insig-2 signaling by siRNA interference against Insig-2 also disrupted the inhibitory effect of this signaling pathway on hepatic de novo triglyceride synthesis. These actions resulted in the accumulation of lipid droplets in hepatocytes and systemic hyperlipidemia. Our study identified CREBH as the first cellular protein that regulates Insig-2a expression. Glucagon activated the CREBH-Insig-2a signaling pathway to inhibit hepatic de novo lipogenesis and prevent the onset of hepatic steatosis and hypertriglyceridemia.


Microfluidic flow-through reactor and 3D Raman imaging for in situ assessment of mineral reactivity in porous and fractured porous media.
An in-depth understanding of dissolution and precipitation of minerals in porous and fractured porous media and the complex feedback on the transport of fluids is essential for various subsurface applications. In this context, we developed a novel non-destructive "lab-on-chip" approach for quantitative in situ assessments of mineralogical changes in porous media. Our experimental approach involves a microfluidic flow-through reactor of reactive homogeneous and heterogeneous (fractured) porous media coupled with high-resolution imaging. Here, the reactive medium consists of compacted celestine grains seeded in a reservoir within the microfluidic chip. This medium reacts with a barium chloride solution injected into the microreactor at a constant flow rate, leading to the dissolution of celestine and growth of barite. Various seeding processes of the mineral grains allow the creation of homogeneous reactive porous media or the introduction of large heterogeneities such as fractures. Hence, our approach enables high-resolution investigations of reactive transport in fractured porous media. The use of confocal Raman spectroscopic techniques enables the spatio-temporal visualization of the mineral transformation at the pore-scale in two- and three-dimensions. Moreover, advanced pore-scale modelling correlates the hydrological heterogeneities to the geochemical observations in the micro-reactor, which explains the observed discrepancies between homogeneous and heterogeneous reactive media. Eventually, the proposed methodology can be applied to other chemical systems to provide new insights into hydro-geochemical coupling in porous and fractured porous media as well as high-fidelity datasets to benchmark reactive transport codes that are currently under development.


Diffusive spatio-temporal noise in a first-passage time model for intracellular calcium release.
The intracellular release of calcium from the endoplasmic reticulum is controlled by ion channels. The resulting calcium signals exhibit a rich spatio-temporal signature, which originates at least partly from microscopic fluctuations. While stochasticity in the gating transition of ion channels has been incorporated into many models, the distribution of calcium is usually described by deterministic reaction-diffusion equations. Here we test the validity of the latter modeling approach by using two different models to calculate the frequency of localized calcium signals (calcium puffs) from clustered IP3 receptor channels. The complexity of the full calcium system is here limited to the basic opening mechanism of the ion channels and, in the mathematical reduction simplifies to the calculation of a first passage time. Two models are then studied: (i) a hybrid model, where channel gating is treated stochastically, while calcium concentration is deterministic and (ii) a fully stochastic model with noisy channel gating and Brownian calcium ion motion. The second model utilises the recently developed two-regime method [M. B. Flegg, S. J. Chapman, and R. Erban, "The two-regime method for optimizing stochastic reaction-diffusion simulations," J. R. Soc., Interface 9, 859-868 (2012)] in order to simulate a large domain with precision required only near the Ca(2+) absorbing channels. The expected time for a first channel opening that results in a calcium puff event is calculated. It is found that for a large diffusion constant, predictions of the interpuff time are significantly overestimated using the model (i) with a deterministic non-spatial calcium variable. It is thus demonstrated that the presence of diffusive noise in local concentrations of intracellular Ca(2+) ions can substantially influence the occurrence of calcium signals. The presented approach and results may also be relevant for other cell-physiological first-passage time problems with small ligand concentration and high cooperativity.


Chest palpitations in a teenager as an unusual presentation of Lyme disease: case report.
The incidence of Lyme disease (LD) in North America has increased substantially in the past two decades. Concomitant with the increased incidence of infection has been an enhancement in the recognition of LD complications. Here, we report a case of Lyme carditis complicated by heart block in a pediatric patient admitted to our children's hospital. What is unique about this case is that the complaint of chest palpitations is an infrequent presentation of LD, and what it adds to the scientific literature is an improved understanding of LD in the pediatric population. The patient was a 16-year-old male who presented with the main concerns of acute onset of palpitations and chest pain. An important clinical finding was Erythema migrans (EM) on physical exam. The primary diagnoses were LD with associated Lyme carditis, based on the finding of 1st degree atrioventricular heart block (AVB) and positive IgM and IgG antibodies to Borrelia burgdorferi. Interventions included echocardiography, electrocardiography (EKG), and intravenous antibiotics. The hospital course was further remarkable for transition to 2nd degree heart block and transient episodes of complete heart block. A normal sinus rhythm and PR interval were restored after antibiotic therapy and the primary outcome was that of an uneventful recovery. Lyme carditis occurs in < 5% of LD cases, but the "take-away" lesson of this case is that carditis can be the presenting manifestation of B. burgdorferi infection in pediatric patients. Any patient with suspected Lyme carditis manifesting cardiac symptoms such as syncope, chest pain, or EKG changes should be admitted for parenteral antibiotic therapy and cardiac monitoring. The most common manifestation of Lyme carditis is AVB. AVB may manifest as first-degree block, or may present as high-grade second or third-degree block. Other manifestations of Lyme carditis may include myopericarditis, left ventricular dysfunction, and cardiomegaly. Resolution of carditis is typically achieved through antibiotic administration, although pacemaker placement should be considered if the PR interval fails to normalize or if higher degrees of heart block, with accompanying symptoms, are encountered. With the rising incidence of LD, providers must maintain a high level of suspicion in order to promptly diagnose and treat Lyme carditis.


Pilot testing of a questionnaire for the evaluation of mental health services in family health team clinics in Ontario.
Family health teams (FHTs), regarded today as a premier model of provision of primary care services in North America, were introduced in 2004 to improve traditionally fragmented primary healthcare in Ontario. Physicians and healthcare providers from various disciplines team up under the same roof in FHTs to provide and coordinate care and to ensure adequate access to and continuity of care. Because many Canadians with mental health problems consult family physicians in primary care, routine evaluation of the delivery of primary mental health care services in FHTs is becoming important. The authors' goal was to develop and test an evaluation tool (containing a questionnaire for patients and a questionnaire for providers) for mental health services provided in FHTs with a focus on accessibility, availability, quality, continuity of care and coordination of services. They developed and pilot tested an English-French tailored evaluation instrument in several FHTs in South East, Champlain and North East Local Health Integration Networks across Ontario. A convenience sample of English- and French-speaking healthcare providers and patients using mental health services was recruited. Provider and patient questionnaires were developed and pilot-tested with 12 providers and 10 clients. Patient reviewers rated the patient questionnaire consistently as "good" or "very good." Provider reviewers found the provider questionnaire to be important and timely and the questions to be adequate and interesting. This instrument evaluates, from both the patient and provider perspectives, whether mental health services are structured to meet expectations set for FHTs, and enables healthcare providers, administrators and policy makers to learn about the benefits and the deficiencies of mental health care delivered through these clinics. This instrument can also be used to enhance future research and evaluation of FHTs. Further validation effort will be required to establish its validity and reliability.


Cross-talk between hypoxia and insulin signaling through Phd3 regulates hepatic glucose and lipid metabolism and ameliorates diabetes.
Signaling initiated by hypoxia and insulin powerfully alters cellular metabolism. The protein stability of hypoxia-inducible factor-1 alpha (Hif-1α) and Hif-2α is regulated by three prolyl hydroxylase domain-containing protein isoforms (Phd1, Phd2 and Phd3). Insulin receptor substrate-2 (Irs2) is a critical mediator of the anabolic effects of insulin, and its decreased expression contributes to the pathophysiology of insulin resistance and diabetes. Although Hif regulates many metabolic pathways, it is unknown whether the Phd proteins regulate glucose and lipid metabolism in the liver. Here, we show that acute deletion of hepatic Phd3, also known as Egln3, improves insulin sensitivity and ameliorates diabetes by specifically stabilizing Hif-2α, which then increases Irs2 transcription and insulin-stimulated Akt activation. Hif-2α and Irs2 are both necessary for the improved insulin sensitivity, as knockdown of either molecule abrogates the beneficial effects of Phd3 knockout on glucose tolerance and insulin-stimulated Akt phosphorylation. Augmenting levels of Hif-2α through various combinations of Phd gene knockouts did not further improve hepatic metabolism and only added toxicity. Thus, isoform-specific inhibition of Phd3 could be exploited to treat type 2 diabetes without the toxicity that could occur with chronic inhibition of multiple Phd isoforms.


Evaluation of transport media for Campylobacter jejuni in human fecal specimens.
It is not always possible to culture feces immediately, and appropriate methods for transport of human specimens, unlike those from animals, have not been fully evaluated. Therefore, we took serial subcultures in two phases from six transport media inoculated with human diarrheal stools known to be positive for Campylobacter jejuni. In phase 1, Cary-Blair medium and buffered glycerol saline did not preserve C. jejuni as well as did alkaline peptone-water (APW), modified Cary-Blair medium, thioglycolate broth (Thio), and Campy-Thio. The four best media (APW, Cary-Blair medium, Thio, and Campy-Thio) preserved 20 fecal samples with C. jejuni better at 4 degrees C (90% survival for 5 to 8 days) than at 25 degrees C (90% survival for 1.7 to 2 days). In phase 2, APW and Thio, along with four modifications of the best media in phase 1, were tested with 23 positive strains. The ranges of survival times with modified media at 25 degrees C were 1.3 to 2.2 days (90%) and 4.7 to 6.8 days (50%). APW with reducing agents preserved C. jejuni better than did APW alone, Thio plus ox bile, or Campy-Thio plus ox bile (P less than 0.05). Thio at pH 8.5 was better at preserving C. jejuni than was APW or Thio plus ox bile (P less than 0.05). If human fecal specimens cannot be refrigerated during transport or storage, we recommend the use of Thio at pH 8.5 or APW with reducing agents for preservation of C. jejuni at 25 degrees C.


Tumor regression after systemic administration of tocotrienol entrapped in tumor-targeted vesicles.
The therapeutic potential of tocotrienol, an extract of vitamin E with anti-cancer properties, is hampered by its failure to specifically reach tumors after intravenous administration, without secondary effects on normal tissues. We hypothesize that the encapsulation of tocotrienol-rich fraction (TRF) within vesicles bearing transferrin, whose receptors are overexpressed on many cancer cells, could result in a selective delivery to tumors after intravenous administration. The objectives of this study are therefore to prepare and characterize transferrin-targeted vesicles encapsulating TRF, and to evaluate their therapeutic efficacy in vitro and in vivo. The entrapment of TRF in transferrin-bearing vesicles led to a 3-fold higher TRF uptake and more than 100-fold improved cytotoxicity in A431 (epidermoid carcinoma), T98G (glioblastoma) and A2780 (ovarian carcinoma) cell lines compared to TRF solution. The intravenous administration of TRF encapsulated in transferrin-bearing vesicles led to tumor regression and improvement of animal survival in a murine xenograft model, contrary to that observed with controls. The treatment was well tolerated by the animals. This work corresponds to the first preparation of a tumor-targeted delivery system able to encapsulate tocotrienol. Our findings show that TRF encapsulated in transferrin-bearing vesicles is a highly promising therapeutic system, leading to tumor regression after intravenous administration without visible toxicity.


Automated joint space width quantification of hand and wrist joints: a proof of concept study.
To compare as proof of concept the sensitivity to change of automated quantification of radiographic wrist and hand joint space width (JSW) with scoring JSW according to the Sharp/van der Heijde scoring method (SHS) in two strategy groups of a treat-to-target and tight-control early rheumatoid arthritis (RA) study. Digital radiographs were assessed for JSW changes of 134 patients of the 236 patients participating in the second Computer Assisted Management in Early Rheumatoid Arthritis trial, of whom both baseline and year 2 radiographs were available (year 1 radiographs n=125). Of those 134 patients, 70 started with methotrexate and prednisone (MTX+Pred) and 64 with MTX and placebo (MTX+Plac). JSW change over 1 and 2 years of the hands and wrists was assessed, applying both the joint space narrowing (JSN) subscore of the SHS by 2 readers and the automated assessment with the JSW quantification software 'JSQ'. For both methods, progression of JSW change of the hand and wrist was analysed using linear mixed modelling (dependent variable 'JSW', factor 'strategy group', covariate 'follow-up time in years', interaction term 'strategy group*follow-up time'; radiographs of baseline, year 1 and year 2 were used). For each method the standardised mean difference (SMD) for the change in JSW from baseline to year 2 between the treatment strategies was obtained using a non-parametric method. Patient characteristics of the current subpopulation were similar to those of the whole study population. JSN of the hand and wrist according to SHS at 2 years was present in 16 vs. 23% in the MTX+Pred group vs. the MTX+Plac group. The mean yearly progression rates of JSW change of the hands and wrists using JSQ were -0.00mm (95% confidence interval (CI) -0.01; 0.01) for MTX+Pred vs. -0.02mm (95%CI -0.03; -0.01) for MTX+Plac, p=0.045, and using SHS JSN they were 0.19 units (95%CI 0.09; 0.30) vs. 0.30 units (95%CI 0.14; 0.45) for MTX+Pred vs. MTX+Plac, p=0.271. The SMD for the change from baseline to year 2 between the treatment strategies was 0.37 for JSQ and 0.13 for SHS JSN. In this proof of concept study the yearly progression rate of JSW change of hand and wrist joints, according to the automated JSW quantification software package 'JSQ', was higher in the group initiating MTX+Plac than in the group initiating MTX+Pred. A similar trend was seen with the JSN assessment according to the SHS method of the hand and wrist. However, JSN of the hand and wrist according to SHS, the current gold standard to assess radiographic progression, was seen in only about 20%. Therefore, further studies are needed to conclude firmly that JSQ should be incorporated into quantitative scoring of radiographs in RA.