Primary lysis of eosinophils in severe desquamative asthma.
Primary lysis of eosinophils liberates free eosinophil granules (FEGs) releasing toxic proteins in association with bronchial epithelial injury repair. Eosinophil lysis may be significantly pathogenic. Bronchial mucosal FEGs are associated with uncontrolled asthma, severe asthma, aspirin-sensitive asthma, and lethal asthma. FEGs in the bronchial wall may characterize severe asthma without sputum eosinophilia. Excessive numbers of sputum FEGs occur in severe exacerbations of asthma and are reduced along with clinical improvement. Occurrence of FEGs affects interpretation of other sputum biomarkers including numbers of eosinophils, ECP, and eosinophil-stained macrophages. Thus, eosinophil lysis produces FEGs as bronchial biomarkers of severe asthma. Blood eosinophils in severe asthma seem primed exhibiting a propensity to lyse that is greater the more severe the asthma. Proclivity of blood eosinophils to lyse also distinguished three levels of severity among children with exacerbations of asthma. Numerous FEGs releasing toxic proteins occur in association with grave derangement and shedding of epithelium in severe asthma. Subepithelial FEGs correlate negatively with intact bronchial epithelium in clinically uncontrolled asthma. Significant correlations between sputum ECP, Creola bodies, and severity of asthma exacerbations have also been demonstrated. Hence, eosinophil lysis apparently causes epithelial desquamation in severe asthma. Exaggerated epithelial repair in turn would contribute to inflammatory and remodelling features of severe asthma. Perseverance of FEGs together with maintained disease activity, despite treatment with 'eosinophil-depleting' steroids and anti-IL5 biologicals, agrees with the possibility that eosinophil lysis is worthy target for novel anti-asthma drugs. Priming and lysis of eosinophils, and protein release from FEGs, are regulated and can be targeted. Eosinophil lysis and FEGs belong to the disease picture of severe asthma and need consideration in asthma studies concerned with phenotypes, biomarkers, roles of epithelial injury/repair, and targeting novel drugs.


Role of kinetic intermediates in the folding of leech carboxypeptidase inhibitor.
The oxidative folding and reductive unfolding pathways of leech carboxypeptidase inhibitor (LCI; four disulfides) have been characterized in this work by structural and kinetic analysis of the acid-trapped folding intermediates. The oxidative folding of reduced and denatured LCI proceeds rapidly through a sequential flow of 1-, 2-, 3-, and 4-disulfide (scrambled) species to reach the native form. Folding intermediates of LCI comprise two predominant 3-disulfide species (designated as III-A and III-B) and a heterogeneous population of scrambled isomers that consecutively accumulate along the folding reaction. Our study reveals that forms III-A and III-B exclusively contain native disulfide bonds and correspond to stable and partially structured species that interconvert, reaching an equilibrium prior to the formation of the scrambled isomers. Given that these intermediates act as kinetic traps during the oxidative folding, their accumulation is prevented when they are destabilized, thus leading to a significant acceleration of the folding kinetics. III-A and III-B forms appear to have both native disulfides bonds and free thiols similarly protected from the solvent; major structural rearrangements through the formation of scrambled isomers are required to render native LCI. The reductive unfolding pathway of LCI undergoes an apparent all-or-none mechanism, although low amounts of intermediates III-A and III-B can be detected, suggesting differences in protection against reduction among the disulfide bonds. The characterization of III-A and III-B forms shows that the former intermediate structurally and functionally resembles native LCI, whereas the III-B form bears more resemblance to scrambled isomers.


Primary non-Hodgkin's T-cell lymphoma of the thyroid gland complicating Hashimoto's thyroiditis: case report.
This case report presents an extremely rare case of primary non-Hodgkin's T-cell lymphoma of the thyroid gland complicating Hashimoto's thyroiditis and discusses the clinical history, findings, treatment, and prognosis. Although the place of surgery in the treatment of thyroid lymphoma is controversial, in this case, surgery followed by three rounds of chemotherapy with cyclophosphamide, doxorubicin, vincristine, and prednisone, and radiation therapy to neck and mediastinum were a very effective treatment for the disease so that no relapse has been detected during 3-year follow-up.


Increasing use of intraoperative cholangiogram in Australia: is it evidence-based?
The role of routine intraoperative cholangiograms (IOCs) for prevention of bile duct injury (BDI) is contentious. There are recent reports of limited utility of IOC in preventing BDI. In Australia, IOCs are used more frequently than internationally. This study aimed to evaluate the rate of IOC use in Australia and explore potential changes in practice in light of evolving evidence for the utility of IOC. Data were collated using service item numbers in Medicare Benefits Scheme records on the Australian Government Medicare website, for services claimed between 1 January 2001 and 31 December 2019. These data were used to analyse trends in rates of IOC, cholecystectomy and BDI repair. Data were age-standardized to account for changes in the population over time. The number of IOCs claimed increased by 31.8% and cholecystectomies by 7.0% over the study period. Age-standardized service rates per 100 000 persons increased by 5.5 and 32.6, respectively. Rates of IOC per 100 000 cholecystectomies steadily increased across the study period, while BDI repair rates remained low and erratic. Increasing use of IOC over the last 20 years reflects a trend towards routine rather than selective IOC; however, there is little discernible change in the number of BDIs requiring repair procedures. This suggests that routine IOC use to prevent or minimize BDI is unwarranted. Further investigation is required into the selective IOC use in high-risk patients rather than mandatory use in all patients.


Combined levothyroxine plus liothyronine compared with levothyroxine alone in primary hypothyroidism: a randomized controlled trial.
Standard therapy for patients with primary hypothyroidism is replacement with synthetic thyroxine, which undergoes peripheral conversion to triiodothyronine, the active form of thyroid hormone. Within the lay population and in some medical communities, there is a perception that adding synthetic triiodothyronine, or liothyronine, to levothyroxine improves the symptoms of hypothyroidism despite insufficient evidence to support this practice. To evaluate the benefits of treating primary hypothyroidism with levothyroxine plus liothyronine combination therapy vs levothyroxine monotherapy. Randomized, double-blind, placebo-controlled trial conducted from May 2000 to February 2002 at a military treatment facility that serves active duty and retired military personnel and their family members. The trial included a total of 46 patients aged 24 to 65 years with at least a 6-month history of treatment with levothyroxine for primary hypothyroidism. Patients received either their usual dose of levothyroxine (n = 23) or combination therapy (n = 23), in which their usual levothyroxine dose was reduced by 50 micro g/d and substituted with liothyronine, 7.5 micro g, taken twice daily for 4 months. Scores on a hypothyroid-specific health-related quality-of-life (HRQL) questionnaire, body weight, serum lipid levels, and 13 neuropsychological tests measured before and after treatment. Serum thyrotropin levels remained similar and within the normal range in both treatment groups from baseline to 4 months. Body weight and serum lipid levels did not change. The HRQL questionnaire scores improved significantly in both the control group (23%; P<.001) and the combination therapy group (12%; P =.02), but these changes were statistically similar (P =.54). In 12 of 13 neuropsychological tests, outcomes between groups were not significantly different; the 1 remaining test (Grooved Peg Board) showed better performance in the control group. Compared with levothyroxine alone, treatment of primary hypothyroidism with combination levothyroxine plus liothyronine demonstrated no beneficial changes in body weight, serum lipid levels, hypothyroid symptoms as measured by a HRQL questionnaire, and standard measures of cognitive performance.


Under pressure: The paradox of autonomy and social norms in drug education.
Much contemporary youth drug education operates from the assumption that young people's decisions to use alcohol and other drugs often stem from a misunderstanding that youth consumption is so common as to constitute a 'social norm'. Known as the 'social norms' approach, the stated aim of this form of drug education is to empower young people to avoid consumption by constituting it as uncommon and, therefore, abnormal. Taking the relationship between autonomy and norms as my primary concern, I examine key assumptions of the social norms approach through an analysis of a dataset of 23 'evidence-based' drug education texts currently recommended for use in Australian secondary schools. Drawing on Rasmussen's (2011) analysis of autonomy as a 'paradox' in which young people are compelled to demonstrate their autonomy by submitting to external authority, I argue that drug education constitutes young people's (lack of) autonomy as the key cause of and solution to youth drug use through three strategies: (1) decision-making exercises that position consumption as the result of an inability to make the rational choice; (2) activities that equate drug consumption with succumbing to peer pressure and failing to demonstrate autonomy; and (3) deployment of population level data on youth drug use that constitutes it as atypical. Together these strategies suggest that while drug education often purports to empower young people to make empowered decisions, it operates as a broader social intervention that seeks to produce compliant rather than autonomous subjects.


Risk factors for thyroid abnormalities after neck irradiation for childhood cancer.
Thyroid evaluations were performed in 95 patients who received radiotherapy to the neck region for childhood cancer five to 34 years earlier. Fifty-six patients (61 percent) had at least one abnormality of serum free thyroxine index, serum thyroid-stimulating hormone (thyrotropin), or thyroid palpation. Seven had subnormal free thyroxine index and 40 had elevated thyrotropin concentrations. Thyroidal radiation doses of 3,000 or more rads and lymphangiography independently increased the risk (p less than or equal to 0.01) of an elevated serum thyrotropin concentration (present in 11 percent of patients with neither risk factor, 50 percent of those who underwent lymphangiography and received less than 3,000 rads, 46 percent of those who had 3,000 or more rads and no lymphangiography, and 76 percent of those with both), but duration of follow-up did not. Twenty-six patients had thyroid nodules and six others had diffuse thyroid enlargement. The frequency of palpable abnormalities increased with the follow-up time after radiation (30 percent of patients followed up less than 10 years had abnormalities versus 43 percent of those followed up 10 or more years, p = 0.03), but was not related to the serum thyrotropin level, radiation dose, or lymphangiography. Among 10 patients who had surgery for nodules, three had localized papillary thyroid carcinomas.


Sweat gland reactivity to local thermal stimulation in dysautonomia.
An attempt has been made to elucidate the cause of hyperhidrosis in dysautonomia by testing sweat gland responsiveness to local application of heat. Heat was applied to the volar surface of the forearm with temperatures of 41.5 C and less for one to ten minutes. For each subject, the minimal duration of heating at the lowest temperature capable of evoking sweating was determined (threshold). Thresholds were significantly lower in all subjects with dysautonomia. Since one control subject, the father of a dysautonomia subject, had the highest threshold, these tests do not permit detection of the heterozygote. A reasonable explanation for enhanced sweating reactivity to local heat application is an increased excitatory state of reflex centers. These patients appear permanently on the verge of generalized sweating likely to result from minimal additional stimulation.


Effects of antegrade and retrograde machine perfusion preservation on cardiac function after transplantation in canines.
Most studies investigating machine perfusion preservation for heart transplantation perfuse through the aortic root (antegrade), but the coronary sinus (retrograde) is a potential option. We hypothesized that retrograde machine perfusion provides better functional protection than static storage, while avoiding the potential irregular perfusion seen when aortic insufficiency occurs with antegrade perfusion. Eighteen canine donor hearts were arrested, procured, and stored in modified Celsior solution for 4 hours by using either static storage at 0°C to 4°C (n = 6) or machine perfusion preservation at 5°C via the aortic root (antegrade, n = 6) or coronary sinus (retrograde, n = 6). Lactate and myocardial oxygen consumption were measured in perfused hearts. Hearts were reimplanted and reperfused for 6 hours with hourly function calculated by using the preload recruitable stroke work (PRSW) relation. Myocardial water content was determined at the end of the experiment. Storage lactate levels and myocardial oxygen consumption were comparable in both perfused groups. The PRSW was increased immediately after bypass in the antegrade group (120.6 ± 19.1 mm Hg) compared with the retrograde (75.0 ± 11.3 mm Hg) and static (78.1 ± 10.5 mm Hg) storage groups (P < .05). At the end of reperfusion, PRSW was higher in the retrograde group (69.8 ± 7.4 mm Hg) compared with the antegrade (40.1 ± 6.8 mm Hg) and static (39.9 ± 10.9 mm Hg) storage groups (P < .05). Myocardial water content was similar among groups. Both antegrade and retrograde perfusion demonstrated excellent functional preservation, at least equivalent to static storage. Initial function was superior in the antegrade group, but the retrograde hearts displayed better function late after reperfusion. Neither perfused group developed significant edema. Machine perfusion preservation is a promising technique for improving results of cardiac transplantation.


GRIN1 variants associated with neurodevelopmental disorders reveal channel gating pathomechanisms.
N-methyl-d-aspartate (NMDA) receptors are expressed at synaptic sites, where they mediate fast excitatory neurotransmission. NMDA receptors are critical to brain development and cognitive function. Natural variants to the GRIN1 gene, which encodes the obligatory GluN1 subunit of the NMDA receptor, are associated with severe neurological disorders that include epilepsy, intellectual disability, and developmental delay. Here, we investigated the pathogenicity of three missense variants to the GRIN1 gene, p. Ile148Val (GluN1-3b[I481V]), p.Ala666Ser (GluN1-3b[A666S]), and p.Tyr668His (GluN1-3b[Y668H]). Wild-type and variant-containing NMDA receptors were expressed in HEK293 cells and primary hippocampal neurons. Patch-clamp electrophysiology and pharmacology were used to profile the functional properties of the receptors. Receptor surface expression was evaluated using fluorescently tagged receptors and microscopy. Our data demonstrate that the GluN1(I481V) variant is inhibited by the open pore blockers ketamine and memantine with reduce potency but otherwise has little effect on receptor function. By contrast, the other two variants exhibit gain-of-function molecular phenotypes. Glycine sensitivity was enhanced in receptors containing the GluN1(A666S) variant and the potency of pore block by memantine and ketamine was reduced, whereas that for MK-801 was increased. The most pronounced functional deficits, however, were found in receptors containing the GluN1(Y668H) variant. GluN1(Y668H)/2A receptors showed impaired surface expression, were more sensitive to glycine and glutamate by an order of magnitude, and exhibited impaired block by extracellular magnesium ions, memantine, ketamine, and MK-801. These variant receptors were also activated by either glutamate or glycine alone. Single-receptor recordings revealed that this receptor variant opened to several conductance levels and activated more frequently than wild-type GluN1/2A receptors. Our study reveals a critical functional locus of the receptor (GluN1[Y668]) that couples receptor gating to ion channel conductance, which when mutated may be associated with neurological disorder.


Curcumin regulates miR-21 expression and inhibits invasion and metastasis in colorectal cancer.
Curcumin has promising potential in cancer prevention and therapy by interacting with proteins and modifying their expression and activity, which includes transcription factors, inflammatory cytokines and factors of cell survival, proliferation and angiogenesis. miR-21 is overexpressed in many tumours, promoting progression and metastasis. In the present study, we examined the potential of curcumin to regulate miR-21, tumour growth, invasion and in vivo metastasis in colorectal cancer. In Rko and HCT116 cells, we identified two new transcriptional start sites of the miR-21 gene and delineated its promoter region. PMA stimulation induced miR-21 expression via motifs bound with AP-1 (activator protein 1) transcription factors. Curcumin treatment reduced miR-21 promoter activity and expression in a dose-dependent manner by inhibiting AP-1 binding to the promoter, and induced the expression of the tumour suppressor Pdcd4 (programmed cell death protein 4), which is a target of miR-21. Curcumin-treated Rko and HCT116 cells were arrested in the G2/M phase with increasing concentrations. Furthermore, curcumin inhibited tumour growth, invasion and in vivo metastasis in the chicken-embryo-metastasis assay [CAM (chorionallantoic membrane) assay]. Additionally, curcumin significantly inhibited miR-21 expression in primary tumours generated in vivo in the CAM assay by Rko and HCT116 cells (P<0.00006 and P<0.035 respectively). Taken together, this is the first paper to show that curcumin inhibits the transcriptional regulation of miR-21 via AP-1, suppresses cell proliferation, tumour growth, invasion and in vivo metastasis, and stabilizes the expression of the tumour suppressor Pdcd4 in colorectal cancer.


Medical schools seeking new ways to cope with funding cutbacks.
Cuts in government funding mean that Canada's medical schools have to seek new ways to raise funds. Susan Thorne examines some of the ways faculties of medicine are coping with change. In the brave new world of medical education, schools are combining classes for medical students and other health professionals, seeking business alliances, encouraging attendance by full-tuition students from other countries and diversifying revenue bases through new programs, such as McGill's new 5-year MD-MBA degree.


Effective Perturbations of the Amplitude, Gating, and Hysteresis of IK(DR) Caused by PT-2385, an HIF-2α Inhibitor.
PT-2385 is currently regarded as a potent and selective inhibitor of hypoxia-inducible factor-2α (HIF-2α), with potential antineoplastic activity. However, the membrane ion channels changed by this compound are obscure, although it is reasonable to assume that the compound might act on surface membrane before entering the cell´s interior. In this study, we intended to explore whether it and related compounds make any adjustments to the plasmalemmal ionic currents of pituitary tumor (GH3) cells and human 13-06-MG glioma cells. Cell exposure to PT-2385 suppressed the peak or late amplitude of delayed-rectifier K+ current (IK(DR)) in a time- and concentration-dependent manner, with IC50 values of 8.1 or 2.2 µM, respectively, while the KD value in PT-2385-induced shortening in the slow component of IK(DR) inactivation was estimated to be 2.9 µM. The PT-2385-mediated block of IK(DR) in GH3 cells was little-affected by the further application of diazoxide, cilostazol, or sorafenib. Increasing PT-2385 concentrations shifted the steady-state inactivation curve of IK(DR) towards a more hyperpolarized potential, with no change in the gating charge of the current, and also prolonged the time-dependent recovery of the IK(DR) block. The hysteretic strength of IK(DR) elicited by upright or inverted isosceles-triangular ramp voltage was decreased during exposure to PT-2385; meanwhile, the activation energy involved in the gating of IK(DR) elicitation was noticeably raised in its presence. Alternatively, the presence of PT-2385 in human 13-06-MG glioma cells effectively decreased the amplitude of IK(DR). Considering all of the experimental results together, the effects of PT-2385 on ionic currents demonstrated herein could be non-canonical and tend to be upstream of the inhibition of HIF-2α. This action therefore probably contributes to down-streaming mechanisms through the changes that it or other structurally resemblant compounds lead to in the perturbations of the functional activities of pituitary cells or neoplastic astrocytes, in the case that in vivo observations occur.


Preventable drug-related hospital admissions.
To estimate the prevalence of preventable drug-related hospital admissions (PDRAs) and to explore if selected study characteristics affect prevalence estimates. Keyword search of MEDLINE (1966-December 1999), International Pharmaceutical Abstracts (1970-December 1999), and hand search. Two reviewers independently selected studies published in peer-reviewed journals and extracted crude prevalence estimates and study characteristics. Trials had to specifically address consequences of drug therapy requiring hospital admission and include a quantitative preventability assessment. Stratified analysis and meta-regression were used to explore the association between study characteristics and prevalence estimates. Fifteen studies reported a median PDRA prevalence of 4.3% (interquartile range [IQR] 3.1-9.5%). The median preventability rate of drug-related admissions was 59% (IQR 50-73%). No evidence of publication bias related to study size could be determined. Because the individual study results were highly heterogeneous (Cochran's Q = 176, df = 14; p < 0.001), no meta-analytic summary estimate was computed. Stratified analysis suggested an association between prevalence estimates and 3 study characteristics: exclusion of first admissions (readmission studies: average PDRA prevalence of 14.0 %, estimated prevalence OR = 3.7); mean age of admissions >70 (OR = 2.1); and inclusion of "indirect" drug-related morbidity, such as omission errors or therapeutic failure (OR = 1.9). There was little evidence of other associations with prevalence estimates, such as selection of specific hospital units, exclusion/inclusion of planned admissions, country, and specified methods of PDRA case ascertainment. Drug-related morbidity is a significant healthcare problem, and a great proportion is preventable. Study methods in prevalence reports vary and should be considered when interpreting findings or planning future research.


Origin of hepatic very-low-density lipoprotein triacylglycerol: the contribution of cellular phospholipid.
When rat hepatocytes were cultured for 24 h in the absence of exogenous fatty acid, the amount of very-low-density lipoprotein (VLDL) triacylglycerol (TAG) secreted (114 +/- 14 micrograms/mg of cell protein) could not be accounted for by the mass of TAG lost from the cells (29 +/- 6.1 micrograms/mg of cell protein) during this period (n = 12). Of the balance (85 +/- 14 micrograms/mg; 94 +/- 15 nmol/mg), a maximum of only 37 nmol/mg of cell protein of TAG could be accounted for by fatty acids synthesized de novo. When labelled exogenous oleate (initial concentration, 0.75 nM) was present in the culture medium, the net gain in cellular plus VLDL TAG (253 +/- 38 micrograms/mg of cell protein per 24 h) was greater than that contributed by the exogenous fatty acid (155 +/- 18.2 micrograms/mg of cell protein, n = 5). Again, the balance (98.8 +/- 18.2 micrograms/mg of cell protein per 24 h) was too great to be accounted for by fatty acid synthesis de novo. In experiments in which cellular glycerolipids were prelabelled with [9, 10(n)-3H]oleic acid, following removal of the labelled fatty acid, there was a net increase in labelled cellular plus VLDL TAG over the next 24 h. That cellular phospholipids are the source of a substantial part of the excess TAG synthesized is supported by the following evidence. (1) The loss of prelabelled cellular phospholipid during culture was greater than could be accounted for by secretion into the medium. (2) During culture of cells prelabelled with 1,2-di-[l-14C]palmitoyl phosphatidylcholine, a substantial amount of label was secreted as VLDL TAG. (3) In pulse-chase experiments, the kinetics of labelled phospholipid turnover were consistent with conversion into a non-phospholipid pool. The enzymology involved in the transfer of phospholipid fatty acids into TAG is probably complex, but the present results suggest that this pathway may represent an important route by which extracellular fatty acids are channelled into VLDL TAG.


Relative inequalities in geographic distribution of health care resources in Kermanshah province, Islamic Republic of Iran.
This study aimed to evaluate inequalities in the geographical distribution of human and physical resources in the health sector of Kermanshah province, Islamic Republic of Iran. In a retrospective, cross-sectional study, data from the Statistical Centre of Iran were used to calculate inequality measures (Gini coefficient and index of dissimilarity) over the years 2005-11. The highest Gini coefficient for human resources was observed for pharmacists in 2005 (0.75) and the lowest for paramedics in 2010 and 2011 (0.10). The highest indices of dissimilarity were also for pharmacists in 2005 (29%) and paramedics in 2011 (3%). For physical resources, the highest and lowest Gini coefficients were for rehabilitation centres in 2010 (0.59) and health houses in 2011 (0.12) respectively. Generally, inequalities in the distribution of health care resources were lower at the end of the study period, although there was potential for more equitable distribution of pharmacists, specialists, health houses and beds.


Hair growth in patients alopecia areata totalis after treatment with simvastatin and ezetimibe.
Alopecia areata is an autoimmune disorder characterized by the sudden development of a circumscribed patch of non-scarring hair loss on the scalp or any hair-bearing surface.The presentation of this disorder can be hair loss in a single circumscribed patch, complete loss of hair on the scalp (alopecia totalis) or complete loss of hair on the entire body (alopecia universalis). The following cases involve two patients with treatment-refractory alopecias that benefited significantly after treatment with a combination of ezetimibe and simvastatin, in addition to the continuation of intra-lesional corticosteroid injections. In this report, the known immunomodulatory effects of statins in combination with ezetimibe are discussed along with the known histopathologic findings of autoimmune alopecia. Major histocompatibility complex class II (MHC-II) and intracellular adhesion molecule-1 (ICAM-1) appear to be involved in both the immunomodulatory effects of statins and the pathophysiology of autoimmune alopecia.


Influence of environmental and anthropogenic factors on the composition, concentration and spatial distribution of microplastics: A case study of the Bay of Brest (Brittany, France).
The concentration and spatial distribution of microplastics in the Bay of Brest (Brittany, France) was investigated in two surveys. Surface water and sediment were sampled at nine locations in areas characterized by contrasting anthropic pressures, riverine influences or water mixing. Microplastics were categorized by their polymer type and size class. Microplastic contamination in surface water and sediment was dominated by polyethylene fragments (PE, 53-67%) followed by polypropylene (PP, 16-30%) and polystyrene (PS, 16-17%) microparticles. The presence of buoyant microplastics (PE, PP and PS) in sediment suggests the existence of physical and/or biological processes leading to vertical transfer of lightweight microplastics in the bay. In sediment (upper 5 cm), the percentage of particles identified by Raman micro-spectroscopy was lower (41%) than in surface water (79%) and may explain the apparent low concentration observed in this matrix (0.97 ± 2.08 MP kg-1 dry sediment). Mean microplastic concentration was 0.24 ± 0.35 MP m-3 in surface water. We suggest that the observed spatial MP distribution is related to proximity to urbanized areas and to hydrodynamics in the bay. A particle dispersal model was used to study the influence of hydrodynamics on surface microplastic distribution. The outputs of the model showed the presence of a transitional convergence zone in the centre of the bay during flood tide, where floating debris coming from the northern and southern parts of the bay tends to accumulate before being expelled from the bay. Further modelling work and observations integrating (i) the complex vertical motion of microplastics, and (ii) their point sources is required to better understand the fate of microplastics in such a complex coastal ecosystem.


Evaluating primary care research networks: a review of currently available tools.
An increasing number of primary care research networks (PCRNs) are being developed around the world. Despite the fact that they have existed for a long time in some countries, little is known about what they have actually achieved. There is an ongoing debate in the literature about the appropriate framework for the evaluation of PCRNs. Here, we aim to provide an overview of the tools that are currently available for measuring the performance of PCRNs and practices involved in PCRNs or research. We performed electronic searches in bibliographic databases and several additional searches. We composed a checklist to evaluate the design, content, and methodological quality of the tools. We identified 4 tools for the evaluation of PCRNs or the measurement of primary care practices involved in PCRNs or research. The results of our study showed that various methods, areas of interest, dimensions, and indicators for the evaluation of PCRNs have been proposed. However, no generic and validated tool that enables meaningful comparison between different network models has been developed. It is, therefore, time to reflect on the appropriateness and effectiveness of PCRNs and determine the desired outcomes (ends) of PCRNs and how we can best achieve them in the future (means). To open up the "black box" of the effectiveness of the PCRNs, it may be relevant to observe the effects of network and research participation on those involved in networks.


The CD1d-natural killer T cell axis in atherosclerosis.
A key role for 'lipid-sensing' CD1-restricted natural killer T (NKT) cells in the pathogenesis of atherosclerosis has been suggested. However, the biology of NKT cells remains poorly characterized, as in different experimental settings their activation was reported to both stimulate and suppress innate and adaptive immune responses. Most of the data from experimental models suggest that NKT cells are proatherogenic; however, it is debated whether the increase in atherosclerosis observed following NKT cell stimulation is a consequence of the inability to induce functional NKT cells rather than the proatherogenic nature of NKT cells. CD1d-expressing antigen-presenting cells and NKT cells were detected in mouse and human atherosclerotic lesions. Furthermore, several lysophospholipids and glycosphingolipids, known to accumulate in atherosclerotic plaques, are antigenic for human NKT cell clones. Lipid transfer proteins, such as apolipoprotein E and microsomal triglyceride transfer protein, are central to NKT cell responses. All these data suggest a profound relation between lipid metabolism, CD1d-NKT cell axis activation and atherosclerosis. In this review, we summarize the advances and gaps in our knowledge of NKT cell biology in the context of atherosclerosis as well as the possibility of influencing NKT cell polarization toward an atheroprotective phenotype.


Downward economic mobility and preterm birth: an exploratory study of Chicago-born upper class White mothers.
A paucity of published data exists on the factors underlying the relatively poor birth outcome of non-Hispanic White women in the United States. To determine whether downward economic mobility is a risk factor for preterm birth (<37 weeks, PTB) among upper class-born White women. Stratified and multilevel logistic regression analyses were performed on an Illinois transgenerational dataset of non-Hispanic White infants (1989-1991) and their women (1956-1976) with appended US census income information. The study sample was restricted to singleton births of Chicago-born upper-class (defined by early-life residence in affluent neighborhoods) non-Hispanic White women. Upper class-born White women (n = 4,891) who did not experience downward economic mobility by the time of delivery had a PTB rate of 5.4 %. Those women who experienced slight (n = 5,112), moderate (n = 2,158), or extreme (n = 339) downward economic mobility had PTB rates of 6.5, 8.5, and 10.1 %, respectively; RR (95 % CI) = 1.2 (1.0-4.0), 1.6 (1.3-1.9), and 1.9 (1.3-2.6), respectively. Maternal downward economic mobility was also associated with an increased prevalence of biologic, medical, and behavioral risk factors. Interestingly, the relationship between moderate to extreme downward mobility and preterm birth was stronger among former low birth weight (<2500 g, LBW) than non-LBW women: 2.8 (1.4-5.8) versus 1.6 (1.3-1.9), respectively. In multilevel logistic regression models, the adjusted odds ratio of preterm birth for former LBW and non-LBW women who experienced any downward mobility (compared to those women with lifelong upper class status) equaled 2.4 (1.1-5.3) and 1.1 (1.0-1.1), respectively. Downward economic mobility is associated with an increased risk of preterm birth among upper class-born White urban women; this phenomenon is strongest among former low birth weight women.


Risk factors for rhegmatogenous retinal detachment in the submacular surgery trials: SST report No. 22.
To identify risk factors associated with the development of rhegmatogenous retinal detachment (RRD) in patients enrolled in the Submacular Surgery Trials. One thousand fifteen patients with eligible subfoveal neovascular lesions in the study eye were assigned randomly to observation or to surgery. Eyes were examined at 3 months, 6 months, 12 months, and 24 months after enrollment to assess study outcomes and adverse events, including RRDs. Adverse events also were reported at other times as clinical personnel became aware of them. Potential risk factors for the development of RRD in study eyes were evaluated using recursive partitioning and logistic regression analysis. Among 506 eyes assigned to surgery, RRD developed in 44 (8.7%) compared with 4 (0.8%) of 509 eyes assigned to observation. Of the 44 eyes in which RRD developed, 27 had age-related macular degeneration (AMD) and large (>3.5 MPS disk areas) hemorrhagic subfoveal neovascular lesions at baseline and represented 16.1% of all eyes with such lesions assigned to surgery. Eyes with AMD and larger hemorrhagic lesions (>16 MPS disk areas) together with relatively poor visual acuity (best-corrected visual acuity < or =20/1280) had a higher risk of RRD (odds ratio = 6.2, 95% confidence interval: 2.2-16.7) compared with those with smaller lesions and better visual acuity at baseline. Poor visual acuity and very large, predominantly hemorrhagic subfoveal neovascular AMD lesion type were the greatest risk factors for RRD after submacular surgery. Submacular surgery should be undertaken in such eyes with full awareness of the risk of RRD during subsequent follow-up.


Do predators influence the behaviour of bats?
Many aspects of animal behaviour are affected by real-time changes in the risk of predation. This conclusion holds for virtually all taxa and ecological systems studied, but does it hold for bats? Bats are poorly represented in the literature on anti-predator behaviour, which may reflect a lack of nocturnal predators specialized on bats. If bats actually experience a world with minimal anti-predator concerns, then they will provide a unique contrast within the realm of vertebrate ecology. Alternatively, such predator-driven behaviour in bats may not yet be fully understood, given the difficulties in working with these highly mobile and nocturnal animals. We provide a wide-ranging exploration of these issues in bat behaviour. We first cover the basic predator-prey information available on bats, both on potential predators and the ways in which bats might perceive predators and respond to attacks. We then cover work relevant to key aspects of bat behaviour, such as choice of daytime roosts, the nature of sleep and torpor, evening roost departures, moonlight avoidance, landscape-related movement patterns, and habitat selection. Overall, the evidence in favour of a strong influence of predators on bat behaviour is equivocal, with the picture clouded by contradictory results and a lack of information on potential predators and the perception of risk by bats. It seems clear that day-active bats run a considerable risk of being killed by diurnal raptors, which are able to capture bats with relative ease. Thus, bats taking advantage of a pulse of insects just prior to sunset are likely taking risks to gain much-needed energy. Further, the choice of daytime roosts by bats is probably strongly influenced by roost safety. Few studies, however, have directly addressed either of these topics. As a group, insectivorous temperate-zone bats show no clear tendency to avoid apparently risky situations, such as activity on moonlit nights. However, some observations are consistent with the idea that predation risk affects choice of movement paths and feeding areas by temperate-zone bats, as well as the timing of roost departures. The behaviour of tropical bats, on the other hand, seems more generally influenced by predators; this is especially true for tropical nectarivores and frugivores, but also for insectivorous bats. Presumably there are more serious predators on bats in the tropics (e.g. specialized raptors or carnivorous bats), but the identity of these predators is unclear. More information is needed to assess fully the influence of predators on bat behaviour. There is much need for work on the ways in which bats perceive predators via auditory, visual, and olfactory cues, and whether bats have some knowledge of the risks posed by different predators. Also needed is information on how predators attack bats and how bats react to attacking predators. Difficult to obtain, but of critical value, will be information on the nature of the predation risk experienced by bats while away from roosts and during the full darkness of night.


Effect of Detecting and Isolating Clostridium difficile Carriers at Hospital Admission on the Incidence of C difficile Infections: A Quasi-Experimental Controlled Study.
Clostridium difficile infection (CDI) is a major cause of health care-associated infection worldwide, and new preventive strategies are urgently needed. Current control measures do not target asymptomatic carriers, despite evidence that they can contaminate the hospital environment and health care workers' hands and potentially transmit C difficile to other patients. To investigate the effect of detecting and isolating C difficile asymptomatic carriers at hospital admission on the incidence of health care-associated CDI (HA-CDI). We performed a controlled quasi-experimental study between November 19, 2013, and March 7, 2015, in a Canadian acute care facility. Admission screening was conducted by detecting the tcdB gene by polymerase chain reaction on a rectal swab. Carriers were placed under contact isolation precautions during their hospitalization. Changes in HA-CDI incidence level and trend during the intervention period (17 periods of 4 weeks each) were compared with the preintervention control period (120 periods of 4 weeks each) by segmented regression analysis and autoregressive integrated moving average (ARIMA) modeling. Concomitant changes in the aggregated HA-CDI incidence at other institutions in Québec City, Québec (n = 6) and the province of Québec (n = 94) were also examined. Overall, 7599 of 8218 (92.5%) eligible patients were screened, among whom 368 (4.8%) were identified as C difficile carriers. During the intervention, 38 patients (3.0 per 10 000 patient-days) developed an HA-CDI compared with 416 patients (6.9 per 10 000 patient-days) during the preintervention control period (P < .001). There was no immediate change in the level of HA-CDIs on implementation (P = .92), but there was a significant decrease in trend over time of 7% per 4-week period (rate ratio, 0.93; 95% CI, 0.87-0.99 per period; P = .02). ARIMA modeling also detected a significant effect of the intervention, represented by a gradual progressive decrease in the HA-CDI time series by an overall magnitude of 7.2 HA-CDIs per 10 000 patient-days. We estimated that the intervention had prevented 63 of the 101 (62.4%) expected cases. By contrast, no significant decrease in HA-CDI rates occurred in the control groups. Detecting and isolating C difficile carriers was associated with a significant decrease in the incidence of HA-CDI. If confirmed in subsequent studies, this strategy could help prevent HA-CDI.


Vancomycin-resistant enterococci (VRE) and the role of the healthcare worker.
Antibiotic resistance has increased dramatically within the last decade. The spread of antibiotic resistant strains of bacteria has become a threat within hospitals. Vancomycin-resistant enterococci (VRE) has emerged as one of these strains. VRE is a robust microorganism and can survive for long periods of time on environmental surfaces. VRE spreads quickly from patient to patient through contact with health care workers. This strain can increase the mortality rate in immuno-compromised patients. Hospital health care workers have an important role to play in the prevention and control of VRE. Proper, and frequent, hand-washing significantly contribute to preventing and controlling the spread of VRE. Providing health care workers with education and resources is also a key factor. The health belief model helps to explain how to approach and implement changes to practice.


Extracellular proteins organize the mechanosensory channel complex in C. elegans touch receptor neurons.
Specialized extracellular matrix (ECM) is associated with virtually every mechanosensory system studied. C. elegans touch receptor neurons have specialized ECM and attach to the surrounding epidermis. The mec-1 gene encodes an ECM protein with multiple EGF and Kunitz domains. MEC-1 is needed for the accumulation of the collagen MEC-5 and other ECM components, attachment, and, separately, for touch sensitivity. MEC-1 and MEC-5 bind to touch processes uniformly and in puncta. These puncta colocalize with and localize the mechanosensory channel complex in the touch neurons. In turn, the production of the MEC-1 and MEC-5 puncta appears to rely on interactions with the neighboring epidermal tissue. These and other observations lead us to propose that extracellular, but not cytoskeletal, tethering of the degenerin channel is needed for mechanosensory transduction. Additionally, our experiments demonstrate an important role of the ECM in organizing the placement of the channel complex.


Brains over brawn: experience overcomes a size disadvantage in fish social hierarchies.
Life experiences can alter cognitive abilities and subsequent behavior. Here we asked whether differences in experience could affect social status. In hierarchical animal societies, high-ranking males that typically win aggressive encounters gain territories and hence access to mates. To understand the relative contributions of social experience and physical environment on status, we used a highly territorial African cichlid fish species, Astatotilapia burtoni, that lives in a dynamic lek-like social hierarchy. Astatotilapia burtoni males are either dominant or submissive and can switch status rapidly depending on the local environment. Although dominant males are innately aggressive, we wondered whether they modulated their aggression based on experience. We hypothesized that as males mature they might hone their fighting tactics based on observation of other males fighting. We compared males of different ages and sizes in distinctly different physical environments and subsequently tested their fighting skills. We found that a size difference previously thought negligible (<10% body length) gave a significant advantage to the larger opponent. In contrast, we found no evidence that increasing environmental complexity affected status outcomes. Surprisingly, we found that males only a few days older than their opponents had a significant advantage during territorial disputes so that being older compensated for the disadvantage of being smaller. Moreover, the slightly older winners exploited a consistent fighting strategy, starting with lower levels of aggression on the first day that significantly increased on the second day, a pattern absent in younger winners. These data suggest that experience is an advantage during fights for status, and that social learning provides more relevant experience than the physical complexity of the territory.


On expedient properties of common biological score functions for multi-modality, adaptive and 4D dose optimization.
Frequently, radiotherapy treatments are comprised of several dose distributions computed or optimized in different patient geometries. Therefore, the need arises to compute the comprehensive biological effect or physical figure of merit of the combined dose of a number of distinct geometry instances. For that purpose the dose is typically accumulated in a reference geometry through deformation fields obtained from deformable image registration. However, it is difficult to establish precise voxel-by-voxel relationships between different anatomical images in many cases. In this work, the mathematical properties of commonly used score functions are exploited to derive an upper boundary for the maximum effect for normal tissue and a lower boundary for the minimum effect for the target of accumulated doses on multiple geometry instances.


Articulation in early and late bilinguals' two languages: evidence from functional magnetic resonance imaging.
The network of cortical and subcortical regions that contribute to articulation was examined in bilinguals using functional magnetic resonance imaging. Participants were all fluent in French and English: half were bilingual from birth and half were 'late bilinguals' who had learned French after the age of 12. Overt articulation resulted in the bilateral activation of the motor cortex, basal ganglia and cerebellum, and also the supplementary motor area, independent of the language spoken. Furthermore, the threshold and extent of the network involved in articulation was identical for the two bilingual groups with the exception of greater variation in the left putamen for the late bilinguals. These data challenge claims that age of acquisition results in fundamental differences in the neural substrates that subserve language in bilinguals.


Flexibility and governance in eukaryotic DNA replication.
Eukaryotic DNA replication begins at numerous but often poorly characterized sequences called origins, which are distributed fairly regularly along chromosomes. The elusive and idiosyncratic nature of origins in higher eukaryotes is now understood as resulting from a strong epigenetic influence on their specification, which provides flexibility in origin selection and allows for tailoring the dynamics of chromosome replication to the specific needs of cells. By contrast, the factors that assemble in trans to make these origins competent for replication and the kinases that trigger initiation are well conserved. Genome-wide and single-molecule approaches are being developed to elucidate the dynamics of chromosome replication. The notion that a well-coordinated progression of replication forks is crucial for many aspects of the chromosome cycle besides simply duplication begins to be appreciated.


G protein-coupled estrogen receptor 1-mediated effects in the rat myometrium.
G protein-coupled estrogen receptor 1 (GPER), also named GPR30, has been previously identified in the female reproductive system. In this study, GPER expression was found in the female rat myometrium by reverse transcriptase-polymerase chain reaction and immunocytochemistry. Using GPER-selective ligands, we assessed the effects of the GPER activation on resting membrane potential and cytosolic Ca(2+) concentration ([Ca(2+)](i)) in rat myometrial cells, as well as on contractility of rat uterine strips. G-1, a specific GPER agonist, induced a concentration-dependent depolarization and increase in [Ca(2+)](i) in myometrial cells. The depolarization was abolished in Na(+)-free saline. G-1-induced [Ca(2+)](i) increase was markedly decreased by nifedipine, a L-type Ca(2+) channel blocker, by Ca(2+)-free or Na(+)-free saline. Intracellular administration of G-1 produced a faster and transitory increase in [Ca(2+)](i), with a higher amplitude than that induced by extracellular application, supporting an intracellular localization of the functional GPER in myometrial cells. Depletion of internal Ca(2+) stores with thapsigargin produced a robust store-activated Ca(2+) entry; the Ca(2+) response to G-1 was similar to the constitutive Ca(2+) entry and did not seem to involve store-operated Ca(2+) entry. In rat uterine strips, administration of G-1 increased the frequency and amplitude of contractions and the area under the contractility curve. The effects of G-1 on membrane potential, [Ca(2+)](i), and uterine contractility were prevented by pretreatment with G-15, a GPER antagonist, further supporting the involvement of GPER in these responses. Taken together, our results indicate that GPER is expressed and functional in rat myometrium. GPER activation produces depolarization, elevates [Ca(2+)](i) and increases contractility in myometrial cells.


Expanding Therapeutic Opportunities for Extrapulmonary Neuroendocrine Carcinoma.
Poorly differentiated neuroendocrine carcinomas (PD-NEC) are rare cancers garnering interest as they become more commonly encountered in the clinic. This is due to improved diagnostic methods and the increasingly observed phenomenon of "NE lineage plasticity," whereby nonneuroendocrine (non-NE) epithelial cancers transition to aggressive NE phenotypes after targeted treatment. Effective treatment options for patients with PD-NEC are challenging for several reasons. This includes a lack of targetable, recurrent molecular drivers, a paucity of patient-relevant preclinical models to study biology and test novel therapeutics, and the absence of validated biomarkers to guide clinical management. Although advances have been made pertaining to molecular subtyping of small cell lung cancer (SCLC), a PD-NEC of lung origin, extrapulmonary (EP)-PD-NECs remain understudied. This review will address emerging SCLC-like, same-organ non-NE cancer-like and tumor-type-agnostic biological vulnerabilities of EP-PD-NECs, with the potential for therapeutic exploitation. The hypotheses surrounding the origin of these cancers and how "NE lineage plasticity" can be leveraged for therapeutic purposes are discussed. SCLC is herein proposed as a paradigm for supporting progress toward precision medicine in EP-PD-NECs. The aim of this review is to provide a thorough portrait of the current knowledge of EP-PD-NEC biology, with a view to informing new avenues for research and future therapeutic opportunities in these cancers of unmet need.


The pro- and antiangiogenic effects of statins.
Clinical studies indicate that 3-hydroxyl-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor (statin) therapy has a cardiovascular protective activity that may result from an improvement in endothelial function. Experimental studies have shown that statins protect against ischaemia-reperfusion injury of the heart and stimulate the growth of new blood vessels in ischemic limbs of normocholesterolemic animals. The mechanisms underlying these serum lipid-independent effects of statins are not completely understood, but there is increasing evidence that they improve endothelial function through molecular mechanisms that mediate an increase in endothelium-derived nitric oxide. Recent research has revealed a link between statins and the serine/threonine protein kinase Akt that regulates multiple angiogenic processes in endothelial cells, including the generation of nitrous oxide. In contrast to these data, it has also been reported that higher doses of statins inhibit endothelial cell migration and angiogenesis. Thus, further studies on the actions of statins may lead to the identification of new pharmacological targets for the control of blood vessel growth.


PCA as a practical indicator of OPLS-DA model reliability.
Principal Component Analysis (PCA) and Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) are powerful statistical modeling tools that provide insights into separations between experimental groups based on high-dimensional spectral measurements from NMR, MS or other analytical instrumentation. However, when used without validation, these tools may lead investigators to statistically unreliable conclusions. This danger is especially real for Partial Least Squares (PLS) and OPLS, which aggressively force separations between experimental groups. As a result, OPLS-DA is often used as an alternative method when PCA fails to expose group separation, but this practice is highly dangerous. Without rigorous validation, OPLS-DA can easily yield statistically unreliable group separation. A Monte Carlo analysis of PCA group separations and OPLS-DA cross-validation metrics was performed on NMR datasets with statistically significant separations in scores-space. A linearly increasing amount of Gaussian noise was added to each data matrix followed by the construction and validation of PCA and OPLS-DA models. With increasing added noise, the PCA scores-space distance between groups rapidly decreased and the OPLS-DA cross-validation statistics simultaneously deteriorated. A decrease in correlation between the estimated loadings (added noise) and the true (original) loadings was also observed. While the validity of the OPLS-DA model diminished with increasing added noise, the group separation in scores-space remained basically unaffected. Supported by the results of Monte Carlo analyses of PCA group separations and OPLS-DA cross-validation metrics, we provide practical guidelines and cross-validatory recommendations for reliable inference from PCA and OPLS-DA models.


Mediastinal teratoma in a free-ranging American black bear (Ursus americanus).
A 2.75-yr-old female American black bear (Ursus americanus) was found emaciated and unable to rise. It was euthanized due to the perceived poor prognosis. An approximately 18 x 8 x 10-cm, multiloculated, well-demarcated mass that extended from the bifurcation of the trachea, laterally displacing the lung lobes and caudally displacing the heart. The mass contained fibrous connective tissue, cartilage, bone, nervous tissue, smooth muscle, and a variety of epithelial structures. A mediastinal teratoma was diagnosed based on the microscopic features of the neoplasm.


Clinical correlates to laboratory measures for use in non-contact anterior cruciate ligament injury risk prediction algorithm.
Prospective measures of high knee abduction moment during landing identify female athletes at high risk for non-contact anterior cruciate ligament injury. Biomechanical laboratory measurements predict high knee abduction moment landing mechanics with high sensitivity (85%) and specificity (93%). The purpose of this study was to identify correlates to laboratory-based predictors of high knee abduction moment for use in a clinic-based anterior cruciate ligament injury risk prediction algorithm. The hypothesis was that clinically obtainable correlates derived from the highly predictive laboratory-based models would demonstrate high accuracy to determine high knee abduction moment status. Female basketball and soccer players (N=744) were tested for anthropometrics, strength and landing biomechanics. Pearson correlation was used to identify clinically feasible correlates and logistic regression to obtain optimal models for high knee abduction moment prediction. Clinical correlates to laboratory-based measures were identified and predicted high knee abduction moment status with 73% sensitivity and 70% specificity. The clinic-based prediction algorithm, including (Odds Ratio: 95% confidence interval) knee valgus motion (1.43:1.30-1.59 cm), knee flexion range of motion (0.98:0.96-1.01 degrees ), body mass (1.04:1.02-1.06 kg), tibia length (1.38:1.25-1.52 cm) and quadriceps to hamstring ratio (1.70:1.06-2.70) predicted high knee abduction moment status with C statistic 0.81. The combined correlates of increased knee valgus motion, knee flexion range of motion, body mass, tibia length and quadriceps to hamstrings ratio predict high knee abduction moment status in female athletes with high sensitivity and specificity. Utilization of clinically obtainable correlates with the prediction algorithm facilitates high non-contact anterior cruciate ligament injury risk athletes' entry into appropriate interventions with the greatest potential to prevent injury.


[Telomeres and biological ageing in cardiovascular disease].
The striking variability in the age of onset of and the manifestation/ absence of manifestation of cardiovascular diseases is inadequately explained by conventional risk factors, but may be explained by variation in biological age. Telomere length is possibly a reliable marker of biological age, shorter telomeres reflecting more advanced age. The initial telomere length ofa person is mainly determined by genetic factors. Moreover, the telomere length shortens with each cell division, and exposition to harmful environmental factors also results in shorter telomeres. Leukocytes of patients with atherosclerosis and heart failure display remarkably shorter telomeres compared to leukocytes of healthy subjects of similar age. Conventional cardiovascular risk factors are also associated with telomere length. If telomeres are indeed causally involved in the pathogenesis of cardiovascular disease, this might provide new avenues for future preventive and therapeutic strategies.


Clinical use of cardiac PET/MRI: current state-of-the-art and potential future applications.
Combined PET/MRI is a novel imaging method integrating the advances of functional and morphological MR imaging with PET applications that include assessment of myocardial viability, perfusion, metabolism of inflammatory tissue and tumors, as well as amyloid deposition imaging. As such, PET/MRI is a promising tool to detect and characterize ischemic and non-ischemic cardiomyopathies. To date, the greatest benefit may be expected for diagnostic evaluation of systemic diseases and cardiac masses that remain unclear in cardiac MRI, as well as for clinical and scientific studies in the setting of ischemic cardiomyopathies. Diagnosis and therapeutic monitoring of cardiac sarcoidosis has the potential of a possible 'killer-application' for combined cardiac PET/MRI. In this article, we review the current evidence and discuss current and potential future applications of cardiac PET/MRI.


Endocrines and their relation to influenza virus infection.
Treatment with testosterone increases proliferation of influenza virus as well as protein anabolism. A relative lack of testosterone caused by castration is associated with a diminished rate of virus growth. When protein catabolism is increased by ACTH or cortisone, the rate of virus proliferation decreases. These results suggest the existence of a correlation between alterations of protein metabolism and virus proliferation.


Novel behavioral indicator of explicit awareness reveals temporal course of frontoparietal neural network facilitation during motor learning.
Deficits in sequential motor learning have been observed in many patient populations. Having an understanding of the individual neural progression associated with sequential learning in healthy individuals may provide valuable insights for effective interventions with these patients. Due to individual variability in motor skill acquisition, the temporal course of such learning will be vary, suggesting a need for a more individualized approach. Knowing when a subject becomes aware of movement patterns may provide a marker with which to identify each individual's learning time course. To avoid interfering with the incidental nature of discovery during learning, such an indicator requires an indirect, behaviorally-based approach. In Part I, our study aimed to identify a reliable behavioral indicator predictive of the presence of incidental explicit awareness in a sequential motor learning task. Part II, utilized the predictive indicator and EEG to provide neural validation of perceptual processing changes temporally correlated with the indicator. Results of Part I provide a reliable predictive indicator for the timing of explicit awareness development. Results from Part II demonstrates strong classification reliability, as well as a significant neural correlation with behavior for subjects developing awareness (EXP), not observed with subjects without awareness (NOEXP). Additionally, a temporal correlation of peak activation between neural regions was noted over frontoparietal regions, suggesting that the incidental discovery of motor patterns may involve a facilitative network during awareness development. The proposed indicator provides a tool in which to further examine potential impacts of awareness associated with incidental, or exploratory, motor learning, while the individual nature of the indicator provides a tool for monitoring progress in rehabilitative, exploratory motor learning paradigms.


Novel function for intestinal intraepithelial lymphocytes. Murine CD3+, gamma/delta TCR+ T cells produce IFN-gamma and IL-5.
Intestinal intraepithelial lymphocytes (IEL) from mice are greater than 80% CD3+ T cells and could be separated into four subsets according to expression of CD4 and CD8. In our studies designed to assess the functions of IEL, namely, cytokine production, it was important to initially characterize the various subsets of T cells that reside in IEL. The major subset was CD4-, CD8+ (75% of CD3+ T cells), which contained approximately 45 to 65% gamma/delta TCR+ and 35 to 45% alpha/beta TCR+ T cells. Approximately 7.5% of IEL T cells were CD4-, CD8- (double negative) and gamma/delta+ population. On the other hand, CD4+, CD8+ (double positive) and CD4+, CD8- fractions represented 10% and 7.5% of CD3+ T cells, respectively, which were all alpha/beta TCR+. Inasmuch as CD3+, CD4-, CD8+ T cells are a major subset of IEL which contain both gamma/delta TCR or alpha/beta TCR-bearing cells, the present study was focused on the capability of this subset of IEL T cells to produce the cytokines IFN-gamma and IL-5. Both gamma/delta TCR+ and alpha/beta TCR+ IEL spontaneously produced IFN-gamma and IL-5, although higher frequencies of cytokine spot-forming cells were associated with the alpha/beta TCR+ subset. Approximately 30% of CD8+, gamma/delta TCR+ cells produced both cytokines, whereas approximately 90% of alpha/beta TCR+ T cells produced either IFN-gamma or IL-5. Both gamma/delta TCR+ and alpha/beta TCR+ IEL possessed large quantities of cytokine-specific mRNA, clearly showing that these IEL were programmed for cytokine production. When IEL were activated with anti-gamma/delta or anti-CD8 antibodies, higher numbers of IFN-gamma and IL-5 spot-forming cells were noted. The present study has provided direct evidence that a major function of IEL involves cytokine production, and this is the first evidence that gamma/delta TCR+ cells in IEL possess the capability of producing both IL-5 and IFN-gamma.


Association between Hospital Birth Volume and Maternal Morbidity among Low-Risk Pregnancies in Rural, Urban, and Teaching Hospitals in the United States.
Objectives This study aims to examine the relationship between hospital birth volume and multiple maternal morbidities among low-risk pregnancies in rural hospitals, urban non-teaching hospitals, and urban teaching hospitals, using a representative sample of U.S. hospitals. Study Design Using the 2011 Nationwide Inpatient Sample from 607 hospitals, we identified 508,146 obstetric deliveries meeting low-risk criteria and compared outcomes across hospital volume categories. Outcomes include postpartum hemorrhage (PPH), chorioamnionitis, endometritis, blood transfusion, severe perineal laceration, and wound infection. Results Hospital birth volume was more consistently related to PPH than to other maternal outcomes. Lowest-volume rural (< 200 births) and non-teaching (< 650 births) hospitals had 80% higher odds (adjusted odds ratio [AOR] = 1.80; 95% CI = 1.56-2.08) and 39% higher odds (AOR = 1.39; 95% CI = 1.26-1.53) of PPH respectively, than those in corresponding high-volume hospitals. However, in urban teaching hospitals, delivering in a lower-volume hospital was associated with 14% lower odds of PPH (AOR = 0.86; 95% CI = 0.80-0.93). Deliveries in rural hospitals had 31% higher odds of PPH than urban teaching hospitals (AOR = 1.31; 95% CI = 1.13-1.53). Conclusions Low birth volume was a risk factor for PPH in both rural and urban non-teaching hospitals, but not in urban teaching hospitals, where higher volume was associated with greater odds of PPH.


Dental students, social policy students and learning disability: do differing attitudes exist?
To examine the attitudes of dental students and social policy students towards learning disabilities in order to identify whether attitudinal differences exist and to suggest recommendations in the dental undergraduate curriculum commensurate with Government legislation in the United Kingdom. A cross-sectional survey of all undergraduate dental students at Queen's University, Belfast and all undergraduate social policy students at University of Ulster. A convenience sample of all undergraduate dental and social policy students was obtained. The students completed a questionnaire to assess attitude towards learning disability. The data were analysed using Cronbach's alpha, Student's t-test and analysis of variance (one-way fixed effect model). The level of statistical significance was set at 5%. The response rate was 83% for dental students and 97% for social policy students. Dental students had significantly lower mean scores and hence less favourable attitudes to learning disability compared with social policy students. Female dental students had significantly higher mean scores and hence more favourable attitudes to learning disability compared with male students. The findings show that dental undergraduates compared with social policy students had less favourable attitudes towards those with learning disability. Dental students should receive training in learning disability and undergraduate programmes should be conceptualised as a spiral curriculum. It is proposed that social policy theory should be introduced into undergraduate dental curricula, that early exposure to learning disability in a community setting should be incorporated into the first undergraduate years and in later undergraduate clinical years students should treat patients with learning disability in order to promote experiential learning and reflective practice.


Mechanisms of pertussis toxin-induced myelomonocytic cell adhesion: role of Mac-1(CD11b/CD18) and urokinase receptor (CD87).
Stimulation of monoblastic U937 cells with transforming growth factor beta 1 and 1,25-(OH)2 vitamin D3 (TGF-beta 1/D3) upregulates urokinase receptor (uPAR) and confers urokinase-dependent adhesiveness to the cells for serum- or vitronectin-coated surfaces. Recent studies show that uPAR itself is a high-affinity adhesion receptor for vitronectin and that urokinase (uPA) is an activator of this adhesive function. In the course of exploring possible G-protein involvement in this adhesion it was observed that TGF-beta 1/D3-primed U937 cells became adhesive to vitronectin in an uPAR-dependent manner when exposed to pertussis toxin (PTX). The adherent response is concentration- and time-dependent, and was not due to the ADP-ribosyltransferase activity of the toxin because the purified B-subunit of PTX was equally effective. Although promoting adhesion to serum- or vitronectin-coated surfaces, PTX blocked spontaneous cell adhesion to fibrinogen, an endogenous ligand for the Mac-1 receptor (CD11b/CD18). Flow cytometry study showed that expression of the alpha-subunit of Mac-1 (CD11b) on primed cells was increased by nearly threefold. Monoclonal antibody to CD11b abolished the PTX-induced cell adhesion and the binding of the primed cells to PTX-coated plates. Activation of Mac-1 receptor by its endogenous ligand fibrinogen induced cell adherent response similar to PTX. PTX, but not uPA, triggered a rapid rise in [Ca2+]i in primed U937 cells, and PTX-induced adhesion was significantly attenuated by 1,2-bis-(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid/acetoxy-methyl ester (BAPTA/AM), a selective membrane-permeant [Ca2+]i chelator. PTX-induced cell adhesion was also prevented by antibodies to uPAR and by conditioned medium containing soluble uPAR. Together these data indicate that PTX B-subunit may bind to Mac-1 integrin, which leads to a rapid rise in [Ca2+]i and subsequent activation of uPAR for adherence to vitronectin, suggesting a functional link between Mac-1 and activation of uPAR important to cellular trafficking and host defence in response to Bordetella pertussis infection.


Immunosuppressive Effects of Annona muricata L. Leaf Extract on Cellular and Humoral Immune Responses in Male Wistar Rats.
Annona muricata</i> L. (Annonaceae) (AM)'s remarkable anti-inflammatory and anti-cancer activities make it a targeted plant to be explored for its immunomodulatory properties. Traditional practitioners have employed various components of AM to cure a variety of ailments, including cancer, diabetes, and inflammation. The present study evaluated the immunosuppressive effects of 80% ethanol extract of of AM leaves in male Wistar rats on different parameters of humoral and cellular immune responses. AM leaf extract (AMLE) was analyzed using UHPLC-MS/MS to profile its secondary metabolites. AMLE was rich in polyphenols which include (epi)catechin-(epi)catechin-(epi) catechin, caffeic acid, coumaroylquinic acid, hyperin, kaempferol, quinic acid and rutin. The rats were administered 100, 200 and 400 mg/kg bw of the extract daily for 14 days. The effects of AMLE on innate immune responses were determined by evaluating phagocytosis, neutrophils migration, reactive oxygen species (ROS) release, CD11b/CD18 integrin expression, and ceruloplasmin, lysozyme and myeloperoxidase (MPO) levels. The adaptive immune parameters were evaluated by immunizing the rats with sheep red blood cells (sRBC) on day 0 and administered orally with AMLE for 14 days. AMLE established significant immunosuppressive effects on the innate immune parameters by inhibiting the neutrophil migration, ROS production, phagocytic activity and expression of CD11b/CD18 integrin in a dose-dependent pattern. AMLE also suppressed ceruloplasmin, MPO and lysozyme expressions in the rat plasma dose-dependently. AMLE dose-dependently inhibited T and B lymphocytes proliferation, Th1 and Th2 cytokine production, CD4+ and CD8+ co-expression in splenocytes, immunoglobulins (IgM and IgG) expression and the sRBC-induced swelling rate of rat paw in delayed-type hypersensitivity (DTH). The strong inhibitory effects on the different parameters of humoral and cellular responses indicate that AMLE has potential to be an important source of effective immunosuppressive agents.


Knowledge of and attitude toward attention-deficit/ hyperactivity disorder (ADHD) assessed among primary schoolteachers of Makkah City in Saudi Arabia.
One of the most prevalent pediatric medical conditions is attention-deficit/hyperactivity disorder (ADHD). It is characterized by symptoms of hyperactivity, impulsivity, and/or inattention. The first people to notice a child's ADHD symptoms are typically their teachers, who may then offer data that are typically helpful in its diagnosis. The purpose of this study was to assess teachers' knowledge of ADHD, its causes and etiology, as well as the key demographic factors that influence said knowledge. The study was conducted among schoolteachers of primary schools located in different regions of Makkah City, Kingdom of Saudi Arabia. We used a stratified random sampling technique to collect our sample, with the inclusion criteria being teachers who were working in a primary school and who were directly involved in teaching students. Data analysis was performed using the IBM SPSS Statistics software version 21. We received 335 responses to our questionnaire. The age of the participants ranged from 18 to 59 years, with the mean age being 40.2 ± 13.8 years. About 66.9% of teachers were women. Most of the teachers in this sample had a bachelor's degree (73.4%), and only 21.8% had diplomas. Regarding the years of teaching experience, 34.6% had teaching experience exceeding 20 years, whereas 20.6% had less than five years of teaching experience. Regarding general knowledge of the teachers, (5.4%) of the teachers had a good level of knowledge. Only 63.6% of teachers were aware that ADHD students frequently struggled with task and activity organization, and only 7.5% of teachers had good knowledge of ADHD treatment, compared to 40.9% who had a good awareness of symptoms and diagnosis. Only the teachers' education level was significantly associated with their knowledge level of ADHD: 25% of teachers with a postgraduate degree had an overall good level of knowledge versus 2.7% of those with a diploma (P = 0.006). The findings of this study lead to the conclusion that elementary school instructors have an inadequate understanding of ADHD. Some teachers misunderstand the signs and general facts associated with ADHD, thereby indicating the need for further training in this area.


Smoothly clipped absolute deviation (SCAD) regularization for compressed sensing MRI using an augmented Lagrangian scheme.
Compressed sensing (CS) provides a promising framework for MR image reconstruction from highly undersampled data, thus reducing data acquisition time. In this context, sparsity-promoting regularization techniques exploit the prior knowledge that MR images are sparse or compressible in a given transform domain. In this work, a new regularization technique was introduced by iterative linearization of the non-convex smoothly clipped absolute deviation (SCAD) norm with the aim of reducing the sampling rate even lower than it is required by the conventional l1 norm while approaching an l0 norm. The CS-MR image reconstruction was formulated as an equality-constrained optimization problem using a variable splitting technique and solved using an augmented Lagrangian (AL) method developed to accelerate the optimization of constrained problems. The performance of the resulting SCAD-based algorithm was evaluated for discrete gradients and wavelet sparsifying transforms and compared with its l1-based counterpart using phantom and clinical studies. The k-spaces of the datasets were retrospectively undersampled using different sampling trajectories. In the AL framework, the CS-MRI problem was decomposed into two simpler sub-problems, wherein the linearization of the SCAD norm resulted in an adaptively weighted soft thresholding rule with a sparsity enhancing effect. It was demonstrated that the proposed regularization technique adaptively assigns lower weights on the thresholding of gradient fields and wavelet coefficients, and as such, is more efficient in reducing aliasing artifacts arising from k-space undersampling, when compared to its l1-based counterpart. The SCAD regularization improves the performance of l1-based regularization technique, especially at reduced sampling rates, and thus might be a good candidate for some applications in CS-MRI.


Open reduction internal fixation vs non-operative management in proximal humerus fractures: a prospective, randomized controlled trial protocol.
Proximal humerus fractures are the third most common fracture in the elderly population and are expected to increase due to the aging population. Surgical fixation with locking plate technology has increased over the last decade despite a lack of proven superiority in the literature. Three previous randomized controlled trials have not shown a difference in patient-centered outcomes when comparing non-operative treatment with open reduction and internal fixation. Low patient enrollment and other methodological concerns however limit the generalizability of these conclusions and as a result, management of these fractures remains a controversy. By comparing the functional outcomes of locked plate surgical fixation versus non-operative treatment of displaced three and four-part proximal humerus fractures in the elderly population with a large scale, prospective, multi-centered randomized controlled trial, the optimal management strategy for this common injury may be determined. We will conduct a prospective, single blind randomized controlled parallel arm trial to compare non-operative management of proximal humerus fractures with open reduction and internal fixation using locked plating technology. One-hundred and sixty patients > age 60 with acute 3- or 4- part proximal humerus fractures will be randomized to either open reduction and internal fixation with locked plating technology or non-operative management treatment arms. The primary outcome measure is the Constant Score at 24 months post-operative. Secondary outcome measures include the American Shoulder and Elbow Surgeon's Score (ASES), EuroQol EQ-5D-5 L Health Questionnaire Score, short form PROMIS upper extremity score and IPAQ for the elderly score. Further outcome measures include assessment of the initial classification, displacement and angulation and the quality of surgical reduction via a standard computed tomography (CT) scan; rates of non-union, malunion, arthrosis, osteopenia or other complications including infection, nerve injury, intra-articular screw penetration, reoperation rates and hospital re-admission rates. The results of this trial will provide Level 1 evidence to guide decision-making in the treatment of proximal humerus fractures in the elderly population. ClinicalTrials.gov NCT02362100 . Registered 5 Feb 2015.


Age dependence of spontaneous plaque forming cells in human peripheral blood in Graves disease, rheumatoid arthritis and normal subjects.
A protein A plaque forming cell (PFC) assay has been used to assess immunoglobulin synthesis by peripheral blood lymphocytes (PBL) from normal subjects and patients with Graves' disease and rheumatoid arthritis. In normal subjects the number of PFC detected immediately after lymphocyte isolation showed a marked decrease with increasing age. In addition, PBL from patients with Graves' disease or rheumatoid arthritis contained similar numbers of PFC to age-matched control subjects. These findings suggest that patients with these immunological disorders do not show polyclonal B cell activation.


Resection for Chronic Hepatic Abscess due to Infected Embolization Coils After Liver Injury.
Liver injuries after blunt abdominal trauma are very common. Non-operative approaches to management are now the standard of care for many patients with up to and including grade V liver injuries. However, the long-term complications associated with coil embolization can be challenging to manage. We present the case of a 29-year-old male who presented with a chronic liver abscess which contained the coils following embolization of a grade IV liver injury and the subsequent transhepatic embolization of the pseudoaneurysm. In addition, the patient developed a fistula draining the abscess through the previously placed drain site that traversed the diaphragm. A multidisciplinary discussion was held between trauma surgery, hepatobiliary surgery, thoracic surgery, and interventional radiology to discuss the best treatment plan. The patient subsequently underwent liver resection, fistula tract resection, and diaphragm repair. This case presents a definitive management strategy for these complex patients.


Five types of olfactory receptor neurons in the strawberry blossom weevil Anthonomus rubi: selective responses to inducible host-plant volatiles.
Plants release hundreds of volatiles that are important in the interaction with herbivorous animals, but which odorants are detected by which species? In this study, single receptor neurons on the antenna of the oligophagous strawberry blossom weevil Anthonomus rubi were screened for sensitivity to naturally produced plant compounds by the use of gas chromatography linked to electrophysiological recordings from single cells. The narrow tuning of the neurons was demonstrated by responses solely to a few structurally related sesquiterpenes, aromatics or monoterpene hydrocarbons out of hundreds of plant constituents tested. We present five olfactory receptor neuron types, identified according to one primary odorant i.e. the compound to which the neurons are most sensitive. These odorants, (-)-germacrene D, (-)-beta-caryophyllene, methyl salicylate, E-beta-ocimene and (3E)-4,8-dimethyl-1,3,7-nonatriene, present in the intact strawberry plant, are induced in higher amounts by weevil feeding. This suggests that these compounds can provide information about the presence of conspecifics. We used protocols especially designed to allow comparison with previously investigated species. Striking similarities, but also differences, are demonstrated between receptor neuron specificity in the strawberry weevil and moths.


[Comparative analysis of the immune response to DNA constructions encoding hepatitis C virus nonstructural proteins].
A promising approach to construction of antiviral vaccines consists in activation of cellular immunity with the DNA vaccines. The goal of this work was to evaluate the efficacy of genetic immunization of mice with DNA pcNS3-NS5B encoding five hepatitis C virus (HCV) nonstructural proteins: NS3, NS4A, NS4B, NS5A, and NS5B in comparison with plasmids containing genes of same individual nonstructural proteins. The DNA constructions were injected intramuscularly in DBA mice three times. The humoral immune response was assessed with ELISA; cellular immune response--in blast transformation reaction, by quantitation of CD4+ and CD8+ T cell proliferation using flow cytofluorometry, by intracellular synthesis and secretion of IFN-gamma and IL-2 in ELISpot and ELISA. It was found that the functionally active T cell response was achieved to antigens presenting NS3, NS4, NS5A, and NS5B epitopes of different HCV genotypes in response to pcNS3-NS5B plasmid and was stronger than that to plasmids carrying individual genes. A high proliferation rate of CD4+ T cells, secretion of IL-2 and IFN-gamma, induction of anti-NS3 and anti-NS5B IgG2a were demonstrated. These findings indicate that DNA construction pcNS3-NS5B is one of promising candidates for anti-HCV vaccine developing.


Ways of understanding parental chronic pain: a typology.
Chronic non-cancer pain (CNCP) is a prevalent occurrence and is experienced by adults in their child-rearing years. Communication within the family about parental illness can be formidable, and family members are often uninformed about illness details. To date, there is no research exploring how children and adolescents understand parental chronic pain, a very complex phenomenon, and its related disability. The aim of this study was to develop a substantive theory that describes how adolescents manage the experience of living with a parent suffering with CNCP and environmental factors that help or hinder this process. Grounded theory was used, the focus of which was adolescents' processes of dealing with parental illness in the interpersonal and environmental contexts of their daily lives. A sample of 30 young adults was recruited in northeastern Ohio using theoretical sampling. During open-ended interviews, participants were asked to look back on their adolescence and talk about how they managed living with parental chronic pain. Interview transcripts and field notes were analyzed using constant comparative methods. Six ways of understanding parental chronic pain emerged from the data: noticing something is different, wrestling with not knowing, searching for answers, questioning the validity of pain, developing insight into the complexity of pain, and learning important life lessons. Findings shed light on how adolescents understand and attach meaning and significance to parental chronic pain and disability and serve as the basis for the development of personalized family interventions.


Predictors of future research productivity among ophthalmology residents.
To investigate the characteristics of first-year ophthalmology residents associated with high research productivity in twelve years. All first-year ophthalmology residents in the United States (US) in 2009 listed by institutional websites were identified. Publications were obtained from Scopus and Google Scholar, h-indices were calculated, and medical school characteristics such as research productivity, region, rank, and average USMLE Steps 1 and 2 scores were obtained from National Institutes of Health, US Census Bureau, and US News. Data were obtained on 70% (316/453) of residents; most were male, had Doctor of Medicine (MD) degrees, and graduated from medical schools in the US or Canada. Having an additional graduate degree [OR: 3.05, 95% CI: 1.07-8.67], between 1 and 3 publications [OR: 4.16, 95% CI: 2.22-7.79], or 4 or more publications [OR: 14.27, 95% CI: 3.13-64.94] were associated with higher future research productivity. Graduate degrees and publication count were key predictors of future research productivity among ophthalmology residents in the US.


Hemodynamic, antivasoconstrictor, and antiatherosclerotic effects of calcium antagonists in animal models of atherosclerosis.
Calcium antagonists retard the development of atherosclerosis in cholesterol-fed rabbits and modestly enhance regression after their return to a normal diet. Proliferative lesions following endothelial damage (from, for example, balloon catheter, electrical stimulation) are also diminished. Many mechanisms for these effects have been proposed and their relative importance is not yet clear. However, changes in blood lipid levels do not play an important role. Only a few investigations into how atherosclerosis affects the hemodynamic actions of calcium antagonists have been carried out. Thus, the effects of isradipine were compared in atherosclerotic and normal rabbits. Isradipine increased heart rate and cardiac output less in atherosclerotic rabbits than in normal ones while having no effect on the surface electrocardiogram (ECG). In contrast, the arteriolar vasodilator, dihydralazine, induced ST-segment depression with similar falls in blood pressure, partly explainable by reflex tachycardia and intramyocardial maldistribution of coronary blood flow. Flow to the brain increased with isradipine and decreased with dihydralazine. In atherosclerotic animals, the pressor effects of norepinephrine, phenylephrine, and angiotensin II (Ang II) were amplified. Isradipine partly corrected this enhanced responsiveness. Calcium antagonists thus elicit beneficial hemodynamic and antivasoconstrictor effects in atherosclerotic experimental animals, in addition to having a long-term prophylactic antiatherosclerotic action.


A liquid biopsy signature of circulating extracellular vesicles-derived RNAs predicts response to first line chemotherapy in patients with metastatic colorectal cancer.
Colorectal cancer (CRC) is one of the most threatening tumors in the world, and chemotherapy remains dominant in the treatment of metastatic CRC (mCRC) patients. The purpose of this study was to develop a biomarker panel to predict the response of the first line chemotherapy in mCRC patients. Totally 190 mCRC patients treated with FOLFOX or XEOLX chemotherapy in 3 different institutions were included. We extracted the plasma extracellular vesicle (EV) RNA, performed RNA sequencing, constructed a model and generated a signature through shrinking the number of variables by the random forest algorithm and the least absolute shrinkage and selection operator (LASSO) algorithm in the training cohort (n = 80). We validated it in an internal validation cohort (n = 62) and a prospective external validation cohort (n = 48). We established a signature consisted of 22 EV RNAs which could identify responders, and the area under the receiver operating characteristic curve (AUC) values was 0.986, 0.821, and 0.816 in the training, internal validation, and external validation cohort respectively. The signature could also identify the progression-free survival (PFS) and overall survival (OS). Besides, we constructed a 7-gene signature which could predict tumor response to first-line oxaliplatin-containing chemotherapy and simultaneously resistance to second-line irinotecan-containing chemotherapy. The study was first to develop a signature of EV-derived RNAs to predict the response of the first line chemotherapy in mCRC with high accuracy using a non-invasive approach, indicating that the signature could help to select the optimal regimen for mCRC patients.


Differential modulation of Sonic-hedgehog-induced cerebellar granule cell precursor proliferation by the IGF signaling network.
The molecular mechanisms regulating organ growth and size remain unclear. Sonic hedgehog (SHH) signaling is a major player in the regulation of cerebellar development: SHH is secreted by Purkinje neurons and acts on the proliferation of granule cell precursors (GCPs) in the external germinal layer. These then become postmitotic and form the internal granular layer but do so in the presence of SHH ligand, begging the question of how the proliferative response to SHH signaling is downregulated in differentiating GCPs. Here, we have determined the precise cellular localization of the expression of insulin-like growth factor (IGF) network components in the developing mouse cerebellum and show that this network modulates the proliferative effects of SHH signaling on GCPs. IGF1 and IGF2 are potent mitogens for GCPs and both synergize with SHH in inducing GCP proliferation. Whereas the proliferative activity of IGF1 or IGF2 on GCPs does not require intact SHH signaling, aspects of SHH activity on GCP proliferation require signaling through the IGF receptor 1. Moreover, we find that 3 of the IGF-binding proteins, IGFBP2, IGFBP3 and IGFBP5, inhibit IGF1/2-induced cell proliferation, whereas IGFBP5 also inhibits SHH-induced GCPs proliferation. This novel function of IGFBP5 that we have uncovered demonstrates the exquisite regulation of SHH signaling by different components of the IGF network.


Human leukemia inhibitory factor produced by the ExpressTec method from rice (Oryza sativa L.) is active in human neural stem cells and mouse induced pluripotent stem cells.
Stem cell-based therapy has the potential to treat an array of human diseases. However, to study the therapeutic potential and safety of these cells, a scalable cell culture medium is needed that is free of human or bovine-derived serum proteins. Thus, cost-effective recombinant serum proteins and cytokines are needed to produce such mediums. One such cytokine, leukemia inhibitory factor (LIF), has been shown to be a critical paracrine factor that maintains stem cell pluripotency in murine embryonic stem cells and human naïve stem cells while simultaneously inhibiting differentiation. We recently produced recombinant human LIF (rhLIF) in a rice-based protein expression system known as ExpressTec. (12) We described expression of rice-derived rhLIF and demonstrated its biological equivalency to E. coli-derived rhLIF in traditional and embryonic mouse stem cell systems. Here we describe the expression yield of rice-derived rhLIF and the scale up production capacity. We provide further evidence of the efficacy of rice-derived rhLIF in additional stem cell systems including human neural stem cells and mouse induced pluripotent stem (iPS) cells. The expression level, biological activity, and potential for production at commercial scale of rice-derived rhLIF provides a proof-of-principal for ExpressTec-derived proteins to produce regulatory-friendly, high performance, and dependable stem cell media.


A gas chromatographic determination of residues of eleven insecticides and two metabolites on olive tree leaves.
A gas chromatographic (GC) method was developed and statistically validated for the simultaneous determination of residues of pyrethroid, endosulfan, and organophosphorus insecticides and some of their metabolites on olive tree leaves. Pesticide residues were extracted by static extraction with acetone-dichloromethane. After evaporation of the extract to dryness and redissolution in acetone, the organophosphorus insecticides were determined by GC with nitrogen-phosphorus detection. Another portion of the extract, after solvent change to acetonitrile, was cleaned up on an Alumina-N cartridge and analyzed for insecticides sensitive to electron-capture detection (ECD), i.e., pyrethroids and endosulfan and its metabolite. Recoveries of the organophosphorus insecticides ranged from 80.7 to 93.3% with relative standard deviations (RSDs) of < or = 7.2%; recoveries of the ECD-sensitive insecticides ranged from 71.6 to 89.5% with RSDs of < or = 11.6%. The method was used to analyze 26 samples of olive tree leaves from organic olive groves all over Greece, and the results confirmed the viability of the method for routine analysis. Residues of fenthion and fenthion sulfoxide were found in one and 3 samples, respectively, and their identities were confirmed by GC with mass spectrometry.


Attitudes of the general population towards preconception expanded carrier screening for autosomal recessive disorders including inborn errors of metabolism.
A substantial number of severely debilitating and often ultimately fatal inborn errors of metabolism (IEMs) still lack an effective disease-modifying treatment. Informing couples before a pregnancy about an increased risk of having a child with an inherited disorder is now feasible by preconception expanded carrier screening (ECS). While knowledge about carrier status enhances reproductive autonomy, it may also result in ethical dilemmas. The purpose of this study was to assess the attitudes of the general Dutch population towards preconception ECS and to investigate which factors influence these attitudes. Data collection was carried out in collaboration with a market research agency. In total, 1188 Dutch individuals of reproductive age (18-45 years) were invited by email to complete an online ECS questionnaire in 2016. Prior to the start of the questionnaire, a written explanation of the concepts of autosomal recessive (AR) inheritance, carrier status and ECS was presented. The questionnaire was completed by 781 individuals (65.7%), of whom 31% indicated they would take an ECS test themselves. In addition, 55% agreed that ECS should be offered to all prospective parents. The most frequently selected argument in favor of ECS (47.2%) was that participants want to spare a child from a life with a severe hereditary disorder. The reason most often mentioned not to participate in ECS (48%) was that participants reported not having a hereditary disorder in the family. The majority preferred receiving individual test results above a couple-based disclosure method in which participants receive the carrier status results only when they are a carrier couple of the same disorder. Participants with religious beliefs were less likely to participate in ECS, whereas participants who were considering a (future) pregnancy were more likely to participate. Our study demonstrates an overall positive attitude among participants of reproductive age in the general Dutch population towards preconception ECS. A striking misconception is that many of the participants believe that ECS is of interest only for those with a positive family history of one of the hereditary disorders. This finding emphasizes the importance of providing understandable, balanced information and education to the general public regarding the concepts of inheritance when presenting the option of carrier screening. Our results provide valuable insights that can be used in the debate about the responsible implementation of preconception ECS for AR disorders, including IEMs.