The molecular mechanism of mitochondria autophagy in yeast.
Mitochondria are critical for supplying energy to the cell, but during catabolism this organelle also produces reactive oxygen species that can cause oxidative damage. Accordingly, quality control of mitochondria is important to maintain cellular homeostasis. It has been assumed that autophagy is the pathway for mitochondrial recycling, and that the selective degradation of mitochondria via autophagy (mitophagy) is the primary mechanism for mitochondrial quality control, although there is little experimental evidence to support this idea. Recent studies in yeast identified several mitophagy-related genes and have uncovered components involved in the molecular mechanism and regulation of mitophagy. Similarly, studies of Parkinson disease and reticulocyte maturation reveal that Parkin and Nix, respectively, are required for mitophagy in mammalian cells, and these analyses have revealed important physiological roles for mitophagy. Here, we review the current knowledge on mitophagy, in particular on the molecular mechanism and regulation of mitophagy in yeast. We also discuss some of the differences between yeast and mammalian mitophagy.


Altered Expression of ORAI and STIM Isoforms in Activated Human Cardiac Fibroblasts.
Cardiac fibrotization is a well-known process characteristic of many cardiac pathological conditions. The key element is excessive activation of cardiac fibroblasts, their transdifferentiation into myofibroblasts, increased production, and accumulation of extracellular matrix proteins, resulting in cardiac stiffness. The exact cellular mechanisms and molecular components involved in the process are not fully elucidated, but the SOCE mechanism could play an important role. Its key molecules are the molecular sensor of calcium in ER/SR - STIM and the highly selective calcium channels Orai located in the plasma membrane. This study aims to evaluate selected SOCE-associated genes in the activation of HCF cell culture by several known substances (phenylephrine, isoprenaline) that represent cardiovascular overload. After cell cultivation, cell medium was collected to measure the soluble collagen content. From the harvested cells, qRT-PCR was performed to determine the mRNA levels of the corresponding genes. The activation of cells was based on changes in the relative expression of collagen genes as well as the collagen content in the medium of the cell culture. We detected an increase in the expression of the Orai2 isoform, a change in the Orai1/Orai3 ratio and also an increase in the expression of the STIM2 isoform. These results suggest an increased activation of the SOCE mechanism under stress conditions of fibroblasts, which supports the hypothesis of fibroblast activation in pathological processes by altering calcium homeostasis through the SOCE mechanism.


Changing women's lives: the primary prevention of violence against women.
Significant numbers of women in the United States experience violence and abuse in their intimate relationships that compromise the health and safety of the women and their children. This article discusses the problem of violence against women and outlines strategies for responding to violence and abuse in the primary care setting. Nurses can play a vital role in the primary prevention of abuse by implementing abuse screening and education on the issue of violence against women. This article presents strategies for the primary prevention of abuse and guidelines for mobilizing support for women. Collective prevention strategies aimed at eliminating violence against women at the institutional, community, professional, and political level are highlighted.


Zebrafish TRPA1 channels are required for chemosensation but not for thermosensation or mechanosensory hair cell function.
Transient receptor potential (TRP) ion channels have been implicated in detecting chemical, thermal, and mechanical stimuli in organisms ranging from mammals to Caenorhabditis elegans. It is well established that TRPA1 detects and mediates behavioral responses to chemical irritants. However, the role of TRPA1 in detecting thermal and mechanical stimuli is controversial. To further clarify the functions of TRPA1 channels in vertebrates, we analyzed their roles in zebrafish. The two zebrafish TRPA1 paralogs are expressed in sensory neurons and are activated by several chemical irritants in vitro. High-throughput behavioral analyses of trpa1a and trpa1b mutant larvae indicate that TRPA1b is necessary for behavioral responses to these chemical irritants. However, TRPA1 paralogs are not required for behavioral responses to temperature changes or for mechanosensory hair cell function in the inner ear or lateral line. These results support a role for zebrafish TRPA1 in chemical but not thermal or mechanical sensing, and establish a high-throughput system to identify genes and small molecules that modulate chemosensation, thermosensation, and mechanosensation.


Differential display of genes in normal and hypoplastic fetal murine lungs.
To evaluate lung development at the level of gene expression, a comparison was made between normal and hypoplastic murine fetal lungs by using the mRNA differential display technique. We focused on altered gene expressions at gestational day (Gd) 19 in normal and hypoplastic murine lungs. Hypoplastic fetal lungs were created by gavaging pregnant mice at Gd8 with 25 mg of nitrofen (2,4-dichlorophenyl-p-nitrophenyl ether) [1]. Normal as well as gavaged mice were euthanized by an overdose of halothane at Gd19, and fetuses were removed by laparotomy. Lungs were excised and total RNA was extracted from normal and hypoplastic fetal lungs. Differential display technique was carried out using the RNAimage kit (GenHunter Corp., TN). Each reverse transcription and polymerase chain reaction (RT-PCR) was performed using one specific anchor primer H-T11M (5'HindIII-T11A/C/G3') and one arbitrary primer. We have used a total of 3 different anchor primers and 24 arbitrary primers for each sample. There were 20 differentially expressed cDNA clones, either induced, inhibited, enhanced, or reduced in hypoplastic fetal lungs as compared to normal. Of these, one clone (NL2) with reduced expression in Gd19 hypoplastic lungs had 100% homology with mouse nucleosome assembly protein I gene. Another clone (NT5) with induced expression in hypoplastic lungs is an unknown gene. Further, analyses of Northern blots of lungs from various gestational ages showed that the expression of NT5 was induced in hypoplastic lungs at Gd18, whereas in normal lungs it was first expressed at the neonatal stage and was increasingly expressed into adulthood. There is a single hybridized band, approximately 400 bp long for NT5 message. Dexamethasone induced expression of NT5 in normal Gd14 pseudoglandular lungs cultured for 7 days; however, different growth factors did not. Northern blot hybridization of multiple adult mice tissues showed NT5 expression in the lung, intestine, and spleen. The thyromimetic action of nitrofen and the interactive functional pathways of dexamethasone with T3 are known. Therefore, we suggest that the isolation and characterization of NT5 may provide valuable information on the regulation of lung development.


Contrast dependence of perceptual grouping in brain-damaged patients with visual extinction.
Extinction is manifested in conditions of bilateral simultaneous stimulation, as a failure to detect the stimulus contra-lateral to the side of a cerebral lesion, while the same stimulus is correctly detected there when presented in isolation. The phenomenon is usually interpreted in terms of impaired mobilization of attention from an attended to an unattended object. We have recently shown, using pairs of Gabor patches as stimuli, that pair detection is maximally improved in conditions where the two stimuli presented simultaneously to the two halves of the visual field are co-oriented and co-axial and their location is not too eccentric. Here we add new information by showing that contrast isotropy of the stimulus pair is important in producing this orientation-similarity gain. The further advantage of co-oriented co-linear stimuli over co-oriented parallel (vertical) stimuli was shown exclusively with iso-contrast stimulus pairs, and was significantly enhanced when the contrast level of the stimulus pair was low. Stimulus properties producing reduced extinction seem to correlate with the selectivity pattern and contrast dependence of (a) spatial lateral facilitation observed in psychophysical studies with normal observers, and (b) long-range interactions observed in the primary visual cortex. Thus, two remote visual stimuli seem to be processed as a single object when the corresponding neuronal activities are linked via long-range lateral interactions. The present demonstration of contrast dependency in such processing, strengthens our previous conjecture that even in the presence of significant, extinction producing, parietal damage, the primary visual cortex preserves the capacity to encode, using long-range lateral interactions, an image description in which visual objects are already segregated from background.


HIV-1 DNA in peripheral blood mononuclear cells is strongly associated with HIV-1 disease progression in recently infected West African adults.
To analyze the association between the HIV-1 DNA level in peripheral blood mononuclear cells (PBMCs) and disease progression in recently infected West African adults. HIV-1 DNA levels were measured in the PBMCs of 200 adults in the French National Agency for Research on AIDS and viral Hepatitis (ANRS) 1220 cohort who had recently been infected with HIV-1. The association between baseline HIV-1 DNA levels and disease progression was analyzed using multivariate Cox regression. Disease progression was defined as the occurrence of any of the following outcomes: death, first World Health Organization stage 3-4 event, or CD4 count<200/mm. About 200 participants were followed for a median of 30 months. At baseline, the median time from HIV-1 seroconversion was 9 months, median CD4 T-cell count was 471/mm, median HIV-1 DNA level was 3.0 log10 copies/10 PBMCs, and median plasma HIV-1 RNA level was 4.6 log10 copies/mL. The 5-year probability of remaining free of any outcome was 0.74 [95% confidence interval (CI): 0.61 to 0.83] and 0.36 (95% CI: 0.23 to 0.49) in patients with baseline HIV-1 DNA<or=3.0 and >3.0 log10 copies/10 PBMCs, respectively (P<0.001). The adjusted hazard ratio of disease progression was 2.17 in patients with HIV-1 DNA>3.0 log10 copies/10 PBMCs compared with other patients (95% CI: 1.24 to 3.80, P=0.007). The only other factor associated with progression was follow-up CD4 count (hazard ratio=1.23 per 100 cells/mm decrease; 95% CI: 1.07 to 1.41, P=0.003). PBMC HIV-1 DNA level was strongly associated with HIV-1 disease progression, even after adjusting for HIV-1 RNA and CD4 T-cell count. Further studies should assess whether patients with high HIV-1 DNA levels should start antiretroviral therapy earlier than other patients.


Aseptic Loosening of Porous Metaphyseal Sleeves and Tantalum Cones in Revision Total Knee Arthroplasty: A Systematic Review.
Bone loss often complicates revision total knee arthroplasty (TKA). Management of metaphyseal defects varies, with no clearly superior technique. Two commonly utilized options for metaphyseal defect management include porous-coated metaphyseal sleeves and tantalum cones. A systematic review was conducted according to the international Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. We combined search terms "Total knee arthroplasty" AND/OR "Sleeve," "Cone" as either keywords or medical subject heading (MeSH) terms in multiple databases according to PRISMA recommendations. All retrieved articles were reviewed and assessed using defined inclusion and exclusion criteria. A total of 27 studies (12 sleeves and 15 cones) of revision TKAs were included. In the 12 studies on sleeve implantation in revision TKAs, 1,617 sleeves were implanted in 1,133 revision TKAs in 1,025 patients. The overall rate of reoperation was 110/1,133 (9.7%) and the total rate of aseptic loosening per sleeve was 13/1,617 (0.8%). In the 15 studies on tantalum cone implantation in revision TKAs, 701 cones were implanted into 620 revision TKAs in 612 patients. The overall rate of reoperation was 116/620 (18.7%), and the overall rate of aseptic loosening per cone was 12/701 (1.7%). Rates of aseptic loosening of the two implants were found to be similar, while the rate of reoperation was nearly double in revision TKAs utilizing tantalum cones. Variability in the selected studies and the likely multifactorial nature of failure do not allow for any definitive conclusions to be made. This review elucidates the necessity for additional literature examining revision TKA implants.


Acute Risk Change: An Innovative Measure of Operative Adverse Events and Perioperative Team Performance.
Cardiac surgical risk models predict mortality preoperatively, whereas intensive care unit (ICU) models predict mortality postoperatively. Finding a large difference between the 2 (an acute risk change [ARC]) may reflect an alteration in the status of the patient related to the surgery. An adverse ARC was associated with morbidity and mortality in an Australian population. The aims of this study were to validate ARC in a UK population and to investigate the possible mechanisms behind ARC. This was a retrospective case-control study. Single, high-volume cardiothoracic hospital. Data from 4,842 cardiac surgical patients were collected between 2013 and 2015. None. EuroSCORE was recalibrated to each preceding year's data. ARC was defined as postoperative minus preoperative percentage mortality risk. Association among ARC, morbidity, and mortality was tested. Cases with large adverse ARC (greater than +15%) were compared with cases with large favorable ARC (less than -10%) with regard to intraoperative adverse events, unmeasured patient risk factors, and postoperative events. Adverse ARC was associated with hospital mortality, ICU stay, ICU readmission, renal support, prolonged intubation and return to the operating room (p < 0.001). Intraoperative adverse events occurred in 23 of 33 patients with adverse ARC; however, only 2 of 17 patients with favorable ARC reported adverse events (p < 0.001). Unmeasured risk factors were present in 48% of patients in the adverse ARC group. ARC is a readily available and sensitive marker that correlates strongly with morbidity and mortality. The use of ARC in local and national quality monitoring could identify areas for improvement of the quality of cardiac surgical care.


Depression: the case for a monoamine deficiency.
The monoamine hypothesis of depression predicts that the underlying pathophysiologic basis of depression is a depletion in the levels of serotonin, norepinephrine, and/or dopamine in the central nervous system. This hypothesized pathophysiology appears to be supported by the mechanism of action of antidepressants: agents that elevate the levels of these neurotransmitters in the brain have all been shown to be effective in the alleviation of depressive symptoms. However, intensive investigation has failed to find convincing evidence of a primary dysfunction of a specific monoamine system in patients with major depressive disorders. Understanding of the etiology of depression has been hampered by the absence of direct measurements of monoamines in humans. However, the monoamine depletion paradigm, which reproduces the clinical syndrome, allows a more direct method for investigating the role of monoamines. Results from such studies show that antidepressant responses are transiently reversed, with the response being dependent on the class of antidepressant. In contrast, monoamine depletion does not worsen symptoms in depressed patients not taking medication, nor does it cause depression in healthy volunteers with no depressive illness. In conclusion, it is clear that antidepressant agents in current use do indeed require intact monoamine systems for their therapeutic effect. However, some debate remains as to the precise role that a deficiency in monoamine system(s) may play in depression itself.


Thyroid nodules, thyroid function and dietary iodine in the Marshall islands.
Thyroid nodules have been found to be common in the population of the Marshall Islands. This has been attributed to potential exposure of radioiodines from the nuclear weapons tests on Bikini and Eniwetok between 1946 and 1958. In order to get a full picture of thyroid pathology in the Marshallese population potentially exposed to radioactive fallout we performed a large thyroid screening programme using palpation, high resolution ultrasound and fine needle biopsies of palpable nodules. In addition, various parameters of thyroid function (free T3, free T4, thyroid stimulating hormone [TSH]) and anti-thyroid antibodies were examined in large proportions of the total population at risk. Since dietary iodine deficiency is an established risk factor for thyroid nodules, iodine concentration in urine samples of 362 adults and 119 children was measured as well as the iodine content of selected staple food products. The expected high prevalence of thyroid nodules was confirmed. There was no indication of an increased rate of impaired thyroid function in the Marshallese population. A moderate degree of iodine deficiency was found which may be responsible for some of the increased prevalence of thyroid nodules in the Marshallese population. Studies on the relationship between exposure to radioiodines and thyroid nodules need to take dietary iodine deficiency into account in the interpretation of findings.


Association Between Aluminum Exposure From Vaccines Before Age 24 Months and Persistent Asthma at Age 24 to 59 Months.
To assess the association between cumulative aluminum exposure from vaccines before age 24 months and persistent asthma at age 24 to 59 months. A retrospective cohort study was conducted in the Vaccine Safety Datalink (VSD). Vaccination histories were used to calculate cumulative vaccine-associated aluminum in milligrams (mg). The persistent asthma definition required one inpatient or 2 outpatient asthma encounters, and ≥2 long-term asthma control medication dispenses. Cox proportional hazard models were used to evaluate the association between aluminum exposure and asthma incidence, stratified by eczema presence/absence. Adjusted hazard ratios (aHR) and 95% confidence intervals (CI) per 1 mg increase in aluminum exposure were calculated, adjusted for birth month/year, sex, race/ethnicity, VSD site, prematurity, medical complexity, food allergy, severe bronchiolitis, and health care utilization. The cohort comprised 326,991 children, among whom 14,337 (4.4%) had eczema. For children with and without eczema, the mean (standard deviation [SD]) vaccine-associated aluminum exposure was 4.07 mg (SD 0.60) and 3.98 mg (SD 0.72), respectively. Among children with and without eczema, 6.0% and 2.1%, respectively, developed persistent asthma. Among children with eczema, vaccine-associated aluminum was positively associated with persistent asthma (aHR 1.26 per 1 mg increase in aluminum, 95% CI 1.07, 1.49); a positive association was also detected among children without eczema (aHR 1.19, 95% CI 1.14, 1.25). In a large observational study, a positive association was found between vaccine-related aluminum exposure and persistent asthma. While recognizing the small effect sizes identified and the potential for residual confounding, additional investigation of this hypothesis appears warranted.


PSYCHIATRIC UNITS IN GENERAL HOSPITALS-Problems in Development and Efficient Operation.
An adequate 25-bed psychiatric unit can be housed in a wing of a general hospital. Even more important than physical facilities are competent personnel, to be headed by a chief psychiatrist and a psychiatric nurse supervisor, for the unit. Incorporating teaching facilities into the unit helps to integrate psychiatry into the other disciplines of medicine in a continuing educational program. Having psychiatric units in general hospitals enables many voluntary patients to be treated in early stages of the disorder, with a high proportion of recoveries. Medicolegal aspects and the lack of adequate coverage of mental disorders by voluntary prepayment health plans are serious problems in the economy of a unit. Improved hospital administration, expanded training programs, educational work by local mental health societies and modification of laws on malpractice and commitment will go far to help solve these problems.


Clinical characteristics and organ system involvement in sarcoidosis: comparison of the University of Minnesota Cohort with other cohorts.
Sarcoidosis is a systemic granulomatous disease of unknown etiology. Clinical cohort studies of different populations are important to understand the high variability in clinical presentation and disease course of sarcoidosis. The aim of the study is to evaluate clinical characteristics, including organ involvement, pulmonary function tests, and laboratory parameters, in a sarcoidosis cohort at the University of Minnesota. We compare the organ system involvement of this cohort with other available cohorts. We conducted a retrospective data collection and analysis of 187 subjects with biopsy-proven sarcoidosis seen at a tertiary center. Organ system involvement was determined using the WASOG sarcoidosis organ assessment instrument. Clinical phenotype groups were classified using the Genomic Research in Alpha-1 Antitrypsin Deficiency and Sarcoidosis criteria. Mean subject age at diagnosis was 45.8 ± 12.4, with a higher proportion of males (55.1%), and a higher proportion of blacks (17.1%) compared to the racial distribution of Minnesota residents (5.95%). The majority (71.1%) of subjects required anti-inflammatory therapy for at least 1 month. Compared to the A Case Control Etiologic Study of Sarcoidosis cohort, there was a higher frequency of extra-thoracic lymph node (34.2% vs. 15.2%), eye (20.9% vs. 11.8%), liver (17.6% vs. 11.5%), spleen (20.9% vs. 6.7%), musculoskeletal (9.6% vs. 0.5%), and cardiac (10.7% vs. 2.3%) involvement in our cohort. A multisystem disease with at least five different organs involved was identified in 13.4% of subjects. A restrictive physiological pattern was observed in 21.6% of subjects, followed by an obstructive pattern in 17.3% and mixed obstructive and restrictive pattern in 2.2%. Almost half (49.2%) were Scadding stages II/III. Commonly employed disease activity markers, including soluble interleukin-2 receptor and angiotensin-converting enzyme, did not differ between treated and untreated groups. This cohort features a relatively high frequency of high-risk sarcoidosis phenotypes including cardiac and multiorgan disease. Commonly-utilized serum biomarkers do not identify subpopulations that require or do better with treatment. Findings from this study further highlight the high-variability nature of sarcoidosis and the need for a more reliable biomarker to predict and measure disease severity and outcomes for better clinical management of sarcoidosis patients.


Comparative efficacy of aspirin and acetaminophen in the reduction of fever in children.
Antipyretics should be employed in the pediatric population whenever it is the clinical judgment of the attending physician that fever should be lowered. Aspirin and acetaminophen are equally effective as antipyretics. The efficacy and safety of these two most common antipyretic agents are examined, and various studies with these agents are critically reviewed. Since acetaminophen has a greater margin of safety at antipyretic dosages, it is preferred to aspirin when an anti-inflammatory effect is not required. The efficacy and safety of combination therapy with acetaminophen and aspirin in pediatric patients--or an alternative treatment regimen with both these drugs--warrant investigation.


Changes in joint cartilage aggrecan after knee injury and in osteoarthritis.
To determine the concentrations of aggrecan fragments in synovial fluid from patients with knee joint injury, osteoarthritis (OA), or acute pyrophosphate arthritis (PPA; pseudogout), and to test their relative reactivity with the 846 epitope, a putative marker of cartilage aggrecan synthesis. Samples of knee joint fluid from 385 patients and 9 healthy-knee volunteers were obtained in a cross-sectional study. Study groups were acute PPA/ pseudogout (n = 60), anterior cruciate ligament (ACL) rupture (n = 159), meniscus lesion (n = 129), and primary knee OA (n = 37). The 846 epitope on aggrecan was assayed by competitive solution-phase radioimmunoassay. Aggrecan fragments were assayed by enzyme-linked immunosorbent assay using a monoclonal antibody (1-F21). Cartilage oligomeric matrix protein (COMP), C-propeptide of type II collagen (CPII), bone sialoprotein, matrix metalloproteinases 1 and 3, and tissue inhibitor of metalloproteinases 1 were previously quantified by immunoassays. Reactivity of the 846 epitope was increased in all study groups compared with the reference group, and was highest in patients with primary OA. The median levels (in microg fetal aggrecan equivalents/ml) of the epitope were 0.28 (range 0.24-0.47) in the reference group, 0.48 (range 0.26-1.32) in PPA/pseudogout, 0.61 (range 0.12-2.87) in ACL rupture, 0.53 (range 0.22-3.02) in meniscus lesion, and 0.68 (range 0.31-4.31) in primary OA. The 846 epitope reactivity per microg aggrecan fragments in the joint fluid was higher in late-stage OA than in early-stage OA. Epitope 846 reactivity correlated positively with several markers of matrix turnover, particularly with COMP (r(s) = 0.421) and CPII (r(s) = 0.307). The observed differences in 846 epitope reactivity in synovial fluid, and its concentration in relation to aggrecan and other markers of matrix turnover, were consistent with marked ongoing changes in aggrecan turnover after joint injury and in the development of OA. OA is thus a disease characterized by dynamic changes in tissue macromolecule turnover, which is reflected by measurable changes in aggrecan epitopes in the synovial fluid.


Uncertainty-Aware Principal Component Analysis.
We present a technique to perform dimensionality reduction on data that is subject to uncertainty. Our method is a generalization of traditional principal component analysis (PCA) to multivariate probability distributions. In comparison to non-linear methods, linear dimensionality reduction techniques have the advantage that the characteristics of such probability distributions remain intact after projection. We derive a representation of the PCA sample covariance matrix that respects potential uncertainty in each of the inputs, building the mathematical foundation of our new method: uncertainty-aware PCA. In addition to the accuracy and performance gained by our approach over sampling-based strategies, our formulation allows us to perform sensitivity analysis with regard to the uncertainty in the data. For this, we propose factor traces as a novel visualization that enables to better understand the influence of uncertainty on the chosen principal components. We provide multiple examples of our technique using real-world datasets. As a special case, we show how to propagate multivariate normal distributions through PCA in closed form. Furthermore, we discuss extensions and limitations of our approach.


Recurrent furunculosis in returning travelers: newly defined entity.
Bacterial skin infection is a common dermatologic problem in travelers, which usually resolves without sequela. In contrast, post-travel recurrent furunculosis (PTRF) is a new unique entity of a sequential occurrence of many furuncles seen after returning home from a trip to the Tropics. The objective of this study was to characterize the disease course and possible causes of PTRF. A retrospective study was conducted on a group of young, healthy individuals (16 males and 5 females), who presented with PTRF after returning from tropical countries. In all patients, the first furuncle appeared toward the end of the trip and continued for several months after returning home. The average duration of disease was 8.4 months with an average of 4.2 recurrences. Along the disease course, subsequent recurrences became shorter and milder with longer inter-recurrence intervals. Bacterial cultures most commonly grew methicillin-sensitive Staphylococcus aureus (MSSA, 76.5%). Nasal colonization was demonstrated in 47% of patients. There were neither companion travelers nor family members experiencing furuncles. PTRF should be defined as a clinical entity with prolonged travel to the Tropics being its major risk factor. In the author's opinion, a transient immune change in a subpopulation of travelers ignites a series of recurrent furuncles, resolving upon restoration of normal immunity.


Fetal tissue engineering from amniotic fluid.
We have recently shown, in an animal model, that amniotic fluid can be a source of cells for fetal tissue engineering. This study was aimed at determining whether fetal tissue constructs could also be engineered from cells normally found in human amniotic fluid. Cells obtained from the amniotic fluid of pregnant women at 15 to 19 weeks of gestation (n=6) were cultured in Dulbecco's Modified Eagle's medium (Sigma Chemical, St Louis, MO) containing 20% fetal bovine serum and 5 ng/mL basic fibroblast growth factor in a 95% humidified, 5% CO(2) chamber at 37 degrees C. A subpopulation of morphologically distinct cells was then mechanically isolated from the rest and selectively expanded. The lineage of this subpopulation of amniocytes was determined by immunofluorescent staining with antibodies against standard intermediate filaments and surface antigens. Cell proliferation rates were determined by oxidation assay. After cell expansion, colonies of amniocytes were statically and dynamically seeded onto both unwoven, 1-mm-thick polyglycolic acid polymer scaffold and acellular human dermis for 72 hours. The resulting constructs were analyzed by scanning electron microscopy. Amniocytes stained positively for smooth muscle actin, vimentin, cytokeratin 18, and fibroblast surface protein, and negatively for desmin, cluster of differentiation 31, and von Willebrand's factor (Dako, Carpenteria, CA). These findings are consistent with a mesenchymal, fibroblast-myofibroblast cell lineage. Mesenchymal amniocytes could be rapidly expanded in culture, based on results of the proliferation assay. Scanning electron microscopy of amniocyte constructs revealed dense, confluent layers of cells surrounding the polymer matrices and firm cell adhesion to both PGA and Alloderm (Lifecell Corp, Branchburg, NJ) scaffolds. No evidence of cell death was observed. Subpopulations of fetal mesenchymal cells can be consistently isolated from human amniotic fluid and rapidly expanded in vitro. Human mesenchymal amniocytes attach firmly to both polyglycolic acid polymer and acellular human dermis. The amniotic fluid can be a valuable and practical cell source for fetal tissue engineering.


Switchable self-protected attractions in DNA-functionalized colloids.
Surface functionalization with DNA is a powerful tool for guiding the self-assembly of nanometre- and micrometre-sized particles. Complementary 'sticky ends' form specific inter-particle links and reproducibly bind at low temperature and unbind at high temperature. Surprisingly, the ability of single-stranded DNA to form folded secondary structures has not been explored for controlling (nano) colloidal assembly processes, despite its frequent use in DNA nanotechnology. Here, we show how loop and hairpin formation in the DNA coatings of micrometre-sized particles gives us in situ control over the inter-particle binding strength and association kinetics. We can finely tune and even switch off the attractions between particles, rendering them inert unless they are heated or held together--like a nano-contact glue. The novel kinetic control offered by the switchable self-protected attractions is explained with a simple quantitative model that emphasizes the competition between intra- and inter-particle hybridization, and the practical utility is demonstrated by the assembly of designer clusters in concentrated suspensions. With self-protection, both the suspension and assembly product are stable, whereas conventional attractive colloids would quickly aggregate. This remarkable functionality makes our self-protected colloids a novel material that greatly extends the utility of DNA-functionalized systems, enabling more versatile, multi-stage assembly approaches.


Utilization of an Online Cash-Pay Pharmacy for Topical 5-Fluorouracil Prescriptions in Older Patients.
High cost of prescription medications presents a challenging issue for older patients with multimorbidities. Topical 5-fluorouracil (5-FU) is an effective treatment for actinic keratoses (AK), a highly prevalent condition among elderly populations, but it is often associated with unpredictable retail prices and high out-of-pocket costs. One online pharmacy offers branded prescription medications at fixed, low prices, but it may be less accessible to older patients for numerous reasons. To determine if the number of patients receiving topical 5-FU from an online pharmacy is proportionate to the national data on expected payment types for patients prescribed topical 5-FU for AK. We conducted a cross-sectional study using weighted pooled data from the National Ambulatory Medical Care Survey (NAMCS) on topical 5-FU prescriptions for AK from 2007-2016. Data regarding online pharmacy use were provided by Dermatology.com for the year 2019. Among patients with AK prescribed topical 5-FU, the most prevalent payment source was Medicare (54%) followed by private insurance (40%). On the online pharmacy, the majority of patients had commercial insurance (71%) followed by Medicaid (12%). Data from Dermatology.com are limited. Lower-cost medications from the online pharmacy site may improve adherence and outcomes in older adults and decrease total cost associated with AK treatment. However, the online pharmacy is underutilized by this population. J Drugs Dermatol. 2020;19(4): doi:10.36849/JDD.2020.4690.


Guggulsterone Mediated JAK/STAT and PPAR-Gamma Modulation Prevents Neurobehavioral and Neurochemical Abnormalities in Propionic Acid-Induced Experimental Model of Autism.
Autism spectrum disorder is a neurodevelopmental disorder marked by repetitive behaviour, challenges in verbal and non-verbal communication, poor socio-emotional health, and cognitive impairment. An increased level of signal transducer and activator of transcription 3 (STAT3) and a decreased level of peroxisome proliferator-activated receptor (PPAR) gamma have been linked to autism pathogenesis. Guggulsterone (GST) has a neuroprotective effect on autistic conditions by modulating these signalling pathways. Consequently, the primary objective of this study was to examine potential neuroprotective properties of GST by modulating JAK/STAT and PPAR-gamma levels in intracerebroventricular propionic acid (ICV PPA) induced experimental model of autism in adult rats. In this study, the first 11 days of ICV-PPA injections in rats resulted in autism-like behavioural, neurochemical, morphological, and histopathological changes. The above modifications were also observed in various biological samples, including brain homogenate, CSF, and blood plasma. GST was also observed to improve autism-like behavioural impairments in autistic rats treated with PPA, including locomotion, neuromuscular coordination, depression-like behaviour, spatial memory, cognition, and body weight. Prolonged GST treatment also restored neurochemical deficits in a dose-dependent manner. Chronic PPA administration increased STAT3 and decreased PPAR gamma in autistic rat brain, CSF, and blood plasma samples, which were reversed by GST. GST also restored the gross and histopathological alterations in PPA-treated rat brains. Our results indicate the neuroprotective effects of GST in preventing autism-related behavioural and neurochemical alterations.


Resistance to tobamoviruses in transgenic tobacco plants expressing the coat protein gene of pepper mild mottle virus (Korean isolate).
Red pepper, one of the most important vegetable crops in Korea, is severely affected by viral diseases causing 20-50% reduction in product yield. A pepper strain of tobacco mosaic virus (TMV-p) is the most common virus in red pepper. To study the molecular structure of the TMV-p virus, we generated cDNA clones of the viral genome. Partial sequencing of a few cDNA clones revealed that TMV-p shares a 98% identity at the nucleotide level with the Spanish isolate of pepper mild mottle virus (PMMV-s). This suggests that TMV-p should be reclassified as the Korean isolate of PMMV (PMMV-k). The coat protein (CP) gene together with the 3' untranslated region of the PMMV-k virus was obtained by reverse transcriptase-polymerase chain reaction (RT-PCR) using oligomers deduced from the sequence of PMMV-s. The sequence analysis of the CP gene and the 3' untranslated region further confirmed that PMMV-k is highly related to PMMV-s. The CP gene and the 3' untranslated region of PMMV-k were cloned into a plant expression vector and the construct was introduced into tobacco plants. The transgenic plants expressing the PMMV-k CP gene were delayed in developing systemic disease or failed to develop symptoms at all after inoculation with PMMV-k. Delay of symptoms was also observed when the plants were inoculated with TMV-OM which shares a 74% homology with PMMV-k in the amino acid sequence of the CP region. In a local lesion host, the CP expressing plants exhibited a greatly reduced number of necrotic lesions as compared to control plants after inoculation with TMV-OM. Our results show that CP-mediated viral resistance is readily applicable in the case of PMMV-k and can provide resistance to other viruses in the tobamovirus group.


Antiulcerogenic effect and cytotoxic activity of semi-synthetic crotonin obtained from Croton cajucara Benth.
Trans-dehydrocrotonin, the major diterpene isolated from the bark of Croton cajucara, has good antiulcerogenic activity which, however, is accompanied by toxic effects. On the basis of these results, a semi-synthetic crotonin, named 4SRC, was prepared to determine whether this substance has similar antiulcerogenic activity with lower or no toxicity. The natural crotonin was also isolated from the bark of C. cajucara but was not used due to the small amount obtained. The cytotoxic effect of semi-synthetic crotonin, expressed as cell viability, was assessed in (a) lung fibroblast cell line (V79) derived from Chinese hamsters, a system commonly used for cytotoxicity studies, and (b) rat hepatocytes isolated from male Wistar rats. After treatment, cell viability was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide reduction (MTT reduction), total acid content and neutral red uptake assays. To evaluate V79 cell viability, different concentrations of semi-synthetic crotonin were incubated with the cells. To evaluate the antiulcerogenic effects of semi-synthetic crotonin (50, 100 and 200 mg/kg), we used the models of gastric ulcer induced by ethanol/HCl, stress, indomethacin/bethanechol, and ethanol in male Swiss mice and male Wistar rats. The substance had an IC(50)=500 microM in the neutral red uptake and MTT reduction tests and an IC(50)=200 microM in the nucleic acid content test. With regard to hepatocyte viability after treatment with semi-synthetic crotonin at different concentrations, semi-synthetic crotonin had an IC(50)=10-500 microM in the nucleic acid content and MTT reduction tests and an IC(50)=120 microM in the neutral red uptake test. In another experiment, V79 cells were incubated with the metabolites produced by hepatocytes treated with different concentrations of semi-synthetic crotonin. After a 4-h incubation, semi-synthetic crotonin had an IC(50)=500 microM in the MTT reduction and neutral red uptake tests and an IC(50)=370 microM in nucleic acid content test. The substance had significant antiulcerogenic activity in all models studied, suggesting the presence of a possible antisecretory effect combined with a cytoprotective effect. For this reason, the effect of semi-synthetic crotonin was also evaluated on biochemical parameters of gastric juice and gastric wall mucus, both obtained from pylorus-ligated mice. No significant differences were observed in these parameters between semi-synthetic crotonin-treated and control animals. The results obtained with semi-synthetic crotonin are promising, with a significant preventive effect against gastric ulcer induced by different agents. Our data also show that semi-synthetic crotonin was less toxic than dehydrocrotonin and that the cytotoxic effects decreases with the time that isolated hepatocytes were in culture.


Pleiotropic effect of the SCO2127 gene on the glucose uptake, glucose kinase activity and carbon catabolite repression in Streptomyces peucetius var. caesius.
SCO2127 and SCO2126 (glkA) are adjacent regions located in Streptomyces coelicolor DNA. glkA encodes glucose kinase (Glk), which has been implicated in carbon catabolite repression (CCR) in the genus Streptomyces. In this work, the glkA and SCO2127 genes from S. coelicolor were used, either individually or together, to transform three mutants of Streptomyces peucetius var. caesius resistant to CCR. These mutants present decreased levels of Glk, and deficiency in glucose transport. When the mutants were transformed with a plasmid containing the SCO2127 sequence, glucose uptake and Glk activity values were increased to levels similar to or higher than those of the original strain, and each strain regained sensitivity to CCR. This result was surprising considering that the putative SCO2127 amino acid sequence does not seem to encode a glucose permease or a Glk. In agreement with these results, an increase in glkA mRNA levels was observed in a CCR-resistant mutant transformed with SCO2127 compared with those of the original strain and the CCR-resistant mutant itself. As expected, recombinants containing the glkA sequence reverted Glk to normal activity values, but glucose uptake remained deficient. The data suggest that the SCO2127 gene product enhances transcription of both genes, and support the first specific role for this region in Streptomyces species. The physiological consequence of this effect is an increase in the glucose catabolites that may be involved in eliciting CCR in this genus.


Bombesin: action on gut hormones and calcium in man.
Bombesin, a peptide with widespread biological actions, has been demonstrated in human tissues by immunological methods. To investigate its effect in man, synthetic bombesin was infused at low doses in six male volunteers. Bombesin at 2.4 pmol kg-1 min-1 produced significant rises in plasma insulin, glucagon, pancreatic polypeptide, gastrin, cholecystokinin, motilin, glucose-dependent insulinotropic polypeptide, neurotensin, enteroglucagon, vasoactive intestinal polypeptide, and serum calcium. In contrast, bombesin caused a profound fall in parathyroid hormone levels and reduced plasma glucose concentrations. A late rise in plasma calcitonin was also observed. Bombesin had no significant effect on the pituitary hormones, TSH, GH, PRL, or cortisol. No hormonal changes or alterations in calcium were noted during saline infusions. Bombesin has a marked stimulatory effect on gastrointestinal hormones, which is unique and opposite to the effect of somatostatin, a potent inhibitor of gut hormone release. Bombesin also influences calcium-regulating hormones, either directly or through its action on gut hormones. The bombesin concentrations achieved with the dosages used were low enough to indicate a possible physiological role for the endogenous peptide.


Tumor evolution and intratumor heterogeneity of an oropharyngeal squamous cell carcinoma revealed by whole-genome sequencing.
Head and neck squamous cell carcinoma (HNSCC) is characterized by significant genomic instability that could lead to clonal diversity. Intratumor clonal heterogeneity has been proposed as a major attribute underlying tumor evolution, progression, and resistance to chemotherapy and radiation. Understanding genetic heterogeneity could lead to treatments specific to resistant and metastatic tumor cells. To characterize the degree of intratumor genetic heterogeneity within a single tumor, we performed whole-genome sequencing on three separate regions of an human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma and two separate regions from one corresponding cervical lymph node metastasis. This approach achieved coverage of approximately 97.9% of the genome across all samples. In total, 5701 somatic point mutations (SPMs) and 4347 small somatic insertions and deletions (indels)were detected in at least one sample. Ninety-two percent of SPMs and 77% of indels were validated in a second set of samples adjacent to the discovery set. All five tumor samples shared 41% of SPMs, 57% of the 1805 genes with SPMs, and 34 of 55 cancer genes. The distribution of SPMs allowed phylogenetic reconstruction of this tumor's evolutionary pathway and showed that the metastatic samples arose as a late event. The degree of intratumor heterogeneity showed that a single biopsy may not represent the entire mutational landscape of HNSCC tumors. This approach may be used to further characterize intratumor heterogeneity in more patients, and their sample-to-sample variations could reveal the evolutionary process of cancer cells, facilitate our understanding of tumorigenesis, and enable the development of novel targeted therapies.


Continuous invasive cardiac output monitoring--the Baxter/Edwards Critical-Care Swan Ganz IntelliCath and Viligance system.
We evaluated the Baxter/Edwards Critical-Care Swan Ganz IntelliCath continuous cardiac output catheter and Vigilance continuous cardiac output monitor in critically ill adult intensive care patients, and compared cardiac output measurements obtained from this new system with those from a standard bolus thermodilution technique using cold normal saline. Nine Swan Ganz IntelliCath catheters were inserted into patients selected at random, following the decision that pulmonary artery catheter monitoring was required. A total of 100 comparisons were made in nine patients. We found the Swan Ganz IntelliCath catheter to be both easy to insert and position, and simple to use. There were no complications or technical difficulties. The mean cardiac output measured by the continuous system was 8.00 l/min (SD 1.66/min) compared to 8.02 1/min (SD 1.74 l/min) using bolus techniques. Continuous cardiac output values correlated well with bolus measurements (r = 0.87), there was excellent accuracy (bias of 0.02 l/min) and precision was acceptable (limits of agreement of -1.71 to 1.75 l/min). These larger than anticipated limits of agreement may not reflect any inaccuracy in the continuous measurement system, rather they may highlight the extent of well recognised potential errors inherent in the intermittent bolus technique. The availability of a system to measure cardiac output continuously provides a major improvement in intensive care monitoring. This system will permit the rapid and accurate assessment of the response of patients to therapy while providing increased diagnostic facilities and a new research tool.


Delayed DNA double-strand break repair following platin-based chemotherapy predicts treatment response in head and neck squamous cell carcinoma.
The aim of this study was to investigate if defective repair of DNA double-strand break (DSB) in head and neck squamous cell carcinoma (HNSCC) could be used as an early predictor of treatment response. Tumour biopsy 24-36 h following induction chemotherapy (IC) and pre-treatment biopsies were stained for RAD51 and geminin (S-phase marker) for immunofluorescence in patients with HNSCC. The difference between RAD51 score (percentage of geminin-positive cells that were also positive for RAD51) was calculated for the two specimens. Tumours with a percentage difference of⩽10% were deemed to have repaired IC-induced DSBs, and were classified as 'RAD51 negative'. Response at 3 months post treatment and human papilloma virus (HPV) status were assessed. Thirteen pairs of samples were available for analyses. Three samples were classified as RAD51 negative and 10 as RAD51 positive at 24 h post IC. All of the three patients with tumours classified as RAD51 negative had partial response or progressive disease and the 10 patients with tumours deemed RAD51 positive had a complete response. 100% of the HPV-positive tumours were RAD51 positive and had a complete response. We have demonstrated that impaired DSB DNA repair may underlie enhanced treatment sensitivity of HPV-positive HNSCC and repair capacity following platinum-induced DNA damage predicts response in HNSCC. This has potential as a biomarker for patient selection in trials of DNA damage response pathway modulation.


Testosterone replacement therapy in hypogonadal men: assessing benefits, risks, and best practices.
Hypogonadism is a common condition, especially among older men, but often goes undiagnosed and untreated. It can be associated with a number of signs and symptoms that affect health and quality of life, including feelings of low energy and fatigue; decreased sex drive and performance; decreased muscle mass and strength; decreased bone mineral density; and increased body fat, particularly abdominal fat, a putative risk factor for metabolic syndrome and type 2 diabetes mellitus. The evidence supporting testosterone replacement therapy (TRT) in improving these and related conditions is strong and consistent for body composition and sexual function; moderately consistent for bone mineral density; inconsistent for insulin sensitivity, glycemic control, and lipid profiles; and weak and inconsistent for mood and cognitive function. The concern of some physicians about the potential for TRT to stimulate prostate cancer is not supported by decades of data accumulated to date, though studies of longer duration (eg, 10 years or more) would be even more convincing. Other research needs are discussed. As the front line of health care delivery, primary care physicians need to be vigilant in diagnosing and treating symptomatic hypogonadism. Based on current guidelines, we recommend assessing testosterone levels when an adult man exhibits signs of hypogonadism, and as part of normal medical screening in men starting at age 40 to 50 years, to establish a baseline. A physician should discuss the possibility of TRT with symptomatic patients who have a serum total testosterone level < 300 ng/dL. If TRT is initiated, a patient's response and adverse events should be assessed every 3 to 6 months, and therapy adjusted accordingly.


Topiramate and Metformin Are Effective Add-On Treatments in Controlling Antipsychotic-Induced Weight Gain: A Systematic Review and Network Meta-Analysis.
Background: Antipsychotic drugs may lead to side effects such as obesity, diabetes, dyslipidemia, and cardiovascular disease. The current systematic review and network meta-analysis analyzes and provides an update on the clinical performance of these add-ons in comparison to placebo on body weight and body mass index (BMI) reductions. Methods: A comprehensive literature search was performed on electronic databases: PubMed (1946-), Embase (1974-), Cochrane library (1992-), and OpenGrey (2000-) until 31 July 2018. Network meta-analyses, comparing the body weight change, BMI change and withdrawn due to adverse events of different pharmacological add-ons, was performed using a multivariate meta-regression model with random-effects, adopting a frequentist approach. To rank the prognosis for all add-ons, we used surface under the cumulative ranking (SUCRA) values. Outcomes: From 614 potential studies identified, 27 eligible studies (n = 1,349 subjects) were included. All the studies demonstrated low to moderate risk of bias. For the analysis of body weight change, all add-ons except Ranitidine showed significant weight reductions comparing to placebo. The effectiveness rank based on SUCRA results from highest to lowest was Sibutramine, Topiramate, Metformin, Reboxetine, Ranitidine, and placebo. A similar pattern was seen for BMI change. The analysis of safety outcome did not detect significantly increased withdrawn number from the add-ons. Current evidence showed relatively good tolerance and safety of using the pharmacological add-ons. Interpretation: Topiramate and Metformin are effective add-on treatments in controlling antipsychotic-induced weight gain, comparing to placebo. They are well tolerated in short-term period. Although Sibutramine has the highest rank of the effectiveness, its license has been withdrawn in many countries due to its adverse effects. Hence, Sibutramine should not be adopted to treat antipsychotic-induced weight gain.


Left atrial myxoma. A morphologic and histogenetic study.
Seven cases of excised left atrial myxoma are described. These are compared with organizing and organized thrombi. The tumors occurred at ages ranging from 13 to 48 years with male preponderance. There were distinct histological and histochemical differences between myxomas and thrombi. Myxomas were lobulated, soft and gelatinous. The cells were arranged singly, in syncytium, or in gland like manner and around blood vessels. Scant to abundant acid mucopolysaccharide rich stroma with strong metachromasia was present. The cells on histochemical staining revealed a wide range of differentiation and contained small amounts of neutral mucopolysaccharides. Thrombi, on the other hand, were firm, laminated, and the cells were arranged in several stratified layers. Metachromasia was absent. Neutral polysaccharides were absent within the cells of thrombi, These features support the concept that cardiac myxoma is a neoplasm and arises possibly from multipotential mesenchymal cells.


The stability of insulin solutions in syringes is improved by ensuring lower molecular weight silicone lubricants are absent.
Protein drugs such as insulin are almost universally delivered via glass syringes lubricated with silicone oil. It is not uncommon for prefilled syringes (PFS) to become cloudy, which may affect bioavailability or total drug dose. To examine the role, if any, of the silicone oil lubricant in this process, a systematic evaluation of the degree of insulin denaturation and aggregation as a function of silicone oils of different molecular weights was undertaken. The former was measured using fluorescence changes of aqueous insulin/silicone dispersions, while the latter examined changes in turbidity as a function of mixing and silicone oil type; the results were confirmed at two different insulin concentrations and agitation speeds. Lower molecular weight silicones led to the most rapid denaturation and aggregation, and when examined in blends of silicones at a fixed viscosity of 1000 cSt, commonly used for syringe lubrication, more rapid denaturation/aggregation was noted in blends of silicones containing the largest fractions of low molecular weight materials. As a consequence, the molecular weight profile of silicone lubricants should be established prior to the preparation of prefilled syringes.


Fatigue in Residency Education: Understanding the Influence of Work Hours Regulations in Europe.
Although one proposed solution to the problem of fatigued medical trainees is the implementation of work hours regulations, concerns about the effectiveness of these regulations are growing. Canada remains one of the few Western jurisdictions without legislated regulation. Recent research suggests that fatigue is a complex social construct, rather than simply a lack of sleep; thus, the authors explored how regulations and fatigue are understood in countries with established work hours frameworks to better inform other jurisdictions looking to address trainee fatigue. Using constructivist grounded theory methodology, the authors conducted individual, semistructured interviews in 2015-2016 with 13 postgraduate medical trainees from four European countries with established work hours regulations. Data collection and analysis proceeded iteratively, and the authors used a constant comparative approach to analysis. Trainees reported that they were commonly fatigued and that they violated the work hours restrictions for various reasons, including educational pursuits. Although they understood the regulations were legislated specifically to ensure safe patient care and optimize trainee well-being, they also described implicit meanings (e.g., monitoring for trainee efficiency) and unintended consequences (e.g., losing a sense of vocation). Work hours regulations carry multiple, conflicting meanings for trainees that are captured by three predominant rhetorics: the rhetoric of patient safety, of well-being, and of efficiency. Tensions within each of those rhetorics reveal that managing fatigue within clinical training environments is complex. These findings suggest that straightforward solutions are unlikely to solve the problem of fatigue, assure patient safety, and improve trainee well-being.


Distribution study of Chlamydia trachomatis serovars among high-risk women in China performed using PCR-restriction fragment length polymorphism genotyping.
This was one of the first epidemiological studies in China focused on genital Chlamydia trachomatis serotype distribution in high-risk female populations using omp1 gene-based restriction fragment length polymorphism analysis. One thousand seven hundred seventy cervical swab samples from women attending sexually transmitted disease clinics and female sex workers in six cities in China (Shenzhen and Guangzhou in southern China, Nanjing and Shanghai in eastern China, and Nanning and Chengdu in southwestern China) were subjected to serovar genotyping. The proportion of omp1 genes successfully amplified in 240 C. trachomatis plasmid-positive samples was 94.2% (226/240). Serotypes E (n = 63; 27.9%), F (n = 53; 23.5%), G (n = 28; 12.4%), and D (n = 25; 11.1%) were most prevalent. Though there was no significant difference in the geographic distribution of C. trachomatis, serotype E was predominant in the South (32.1%) and East (27.1%), while serotype F was predominant in the Southwest (28.3%). Serotype F infection was associated with young age and single status. Serovar G was associated with lower abdominal pain; 47.5% of asymptomatic patients were infected with serovar E. These results provide information on distribution of genital C. trachomatis serotypes among high-risk women in China and indicate that high-risk women, including those who are asymptomatic, can be infected with multiple serovars of C. trachomatis, revealing exposure to multiple sources of infection. Although the scope for generalizations is limited by our small sample size, our results showing clinical correlations with genotypes are informative.


Endemic chronic wasting disease causes mule deer population decline in Wyoming.
Chronic wasting disease (CWD) is a fatal transmissible spongiform encephalopathy affecting white-tailed deer (Odocoileus virginianus), mule deer (Odocoileus hemionus), Rocky Mountain elk (Cervus elaphus nelsoni), and moose (Alces alces shirasi) in North America. In southeastern Wyoming average annual CWD prevalence in mule deer exceeds 20% and appears to contribute to regional population declines. We determined the effect of CWD on mule deer demography using age-specific, female-only, CWD transition matrix models to estimate the population growth rate (λ). Mule deer were captured from 2010-2014 in southern Converse County Wyoming, USA. Captured adult (≥ 1.5 years old) deer were tested ante-mortem for CWD using tonsil biopsies and monitored using radio telemetry. Mean annual survival rates of CWD-negative and CWD-positive deer were 0.76 and 0.32, respectively. Pregnancy and fawn recruitment were not observed to be influenced by CWD. We estimated λ = 0.79, indicating an annual population decline of 21% under current CWD prevalence levels. A model derived from the demography of only CWD-negative individuals yielded; λ = 1.00, indicating a stable population if CWD were absent. These findings support CWD as a significant contributor to mule deer population decline. Chronic wasting disease is difficult or impossible to eradicate with current tools, given significant environmental contamination, and at present our best recommendation for control of this disease is to minimize spread to new areas and naïve cervid populations.


A case of manslaughter by heat-stroke.
In the course of his treatment by a herbalist who had undertaken to cure his disability, a mildly spastic retarded four-year-old child was immersed in a heap of fermenting horse manure for 40 minutes. He was unconscious when removed and died two days later despite intensive care in a children's hospital. When medical aid was sought a false history was given and the true nature of the child's illness, heat stroke, was not determined until after death. The herbalist was subsequently convicted of manslaughter.


Methyl (2Z)-2-bromo-meth-yl-3-(3-chloro-phen-yl)prop-2-enoate.
There are two independent mol-ecules (A and B) in the asymmetric unit of the title compound C11H10BrClO2, which represents the Z isomer. The methyl-acrylate moieties are essentially planar, within 0.084 (2) and 0.027 (5) Å in mol-ecules A and B, respectively. The benzene ring makes dihedral angles of 13.17 (7) and 27.89 (9)° with the methyl-acrylate moiety in mol-ecules A and B, respectively. The methyl-bromide moiety is almost orthogonal to the benzene ring, making dihedral angles of 81.46 (16)° in mol-ecule A and 79.61 (16)° in mol-ecule B. The methyl-acrylate moiety exhibits an extended trans conformation in both mol-ecules. In the crystal, pairs of C-H⋯O hydrogen bonds result in the formation of quasi-centrosymmetric R 2 (2)(14) AB dimers.


Let off the leash: kala-azar following the use of tumour necrosis factor antibodies.
A 74-year-old woman with a history of psoriatic arthritis was referred to the Hospital for Tropical Diseases following investigation of a skin lesion that had failed to heal after a visit to Malta 2 years previously. Skin biopsy had revealed invasion of Leishmania amastigotes. She reported a recent history of weight loss, dry cough and dyspnoea, and was investigated for pancytopenia and hepatosplenomegaly. Bone marrow biopsy confirmed the diagnosis of visceral leishmaniasis and she responded well to treatment with intravenous liposomal amphotericin B. Recent rheumatological treatment with adalimumab, a monoclonal antibody to tumour necrosis factor α, was thought to be the factor responsible for causing the cutaneous lesion to become disseminated. This case highlights an unexpected adverse effect of novel immunosuppressants. As the use of biologics becomes widespread, there is an increasing need for clinical surveillance.


[Studies of transgenic tobacco plants expressing beta-1,3-glucanase and chitinase genes and their potential for fungal resistance].
Plant bivalent expression vector pBLGC for basic tobacco beta-1, 3-glucanase and bean chitinase genes were constructed, and the tobacco tissue was transformed. Transgenic tobacco plants were carried out for analysis by PCR, PCR-Southern blot, Dot blot, Western blot, etc., and the results showed that some transgenic plants gave strong positive signal, it suggested that foreign genes have been integrated into tobacco genome, and expressed correctly. Fungal challenge with Alternaria alternata showed that transgenic plants were much more resistant to this pathogenic fungus.


Anxiety and depression among healthcare workers during the COVID-19 pandemic: a systematic umbrella review of the global evidence.
To summarise the evidence relating to the prevalence of anxiety and depression among healthcare workers (HCWs) during the COVID-19 pandemic. An umbrella review of systematic reviews was undertaken using the Joanna Briggs Institute (JBI) methods. The Cochrane database of systematic reviews, JBI Evidence Synthesis, MEDLINE, Web of Science, PsycINFO, Embase and CINAHL were searched in March 2021 for reviews published in English. Systematic reviews reporting the prevalence of anxiety and depression among HCWs during the COVID-19 pandemic. Two researchers screened each abstract and independently reviewed full text articles. Study quality was assessed using the JBI critical appraisal tool for systematic reviews, and the degree of overlap in primary studies was calculated. Ten systematic reviews (100 unique studies), including 169 157 HCWs from 35 countries were included. The prevalence of anxiety among all HCWs ranged from 22.2% (95% CI 21.3% to 23.1%) to 33.0% (95% CI 31.9% to 34.1%). The prevalence of anxiety among physicians (n=5820) was reported to be between 17% and 19.8% and for nurses (n=14 938) between 22.8% and 27%. The prevalence of depression among all HCWs ranged from 17.9% (95% CI 17.1% to 18.8%) to 36% (95% CI 34.9% to 37.1%). The prevalence of depression among physicians (n=643) and nurses (n=8063) was reported to be 40.4% and 28%, respectively. There is wide variation evident in the presence of anxiety and depression among HCWs. In particular, the prevalence of depression among physicians was high. Strategies to reduce the incidence of anxiety and depression are urgently required. CRD42021238960.


A mathematical model of in vivo bovine blastocyst developmental to gestational Day 15.
Bovine embryo growth involves a complex interaction between the developing embryo and the growth-promoting potential of the uterine environment. We have previously established links between embryonic factors (embryo stage, embryo gene expression), maternal factors (progesterone, body condition score), and embryonic growth to 8 d after bulk transfer of Day 7 in vitro-produced blastocysts. In this study we recovered blastocysts on Days 7 and 15 after artificial insemination to test the hypothesis that in vivo and in vitro embryos follow a similar growth program. We conducted our study using 4 commercial farms and repeated our study over 2 yr (2014, 2015), with data available from 2 of the 4 farms in the second year. Morphological and gene expression measurements (196 candidate genes) of the Day 7 embryos were measured and the progesterone concentration of the cows were measured throughout the reproductive cycle as a reflection of the state of the uterine environment. These data were also used to assess the interaction between the uterine environment and the developing embryo and to examine how well Day 7 embryo stage can be predicted from the Day 7 gene expression profile. Progesterone was not a strong predictor of in vivo embryo growth to Day 15. This contrasts with a range of Day 7 embryo transfer studies which demonstrated that progesterone is a very good predictor of embryo growth to Day 15. Our analysis demonstrates that in vivo embryos are 3 times less sensitive to progesterone than in vitro-transferred embryos (up to Day 15). This highlights that caution must be applied when extrapolating the results of in vitro embryo transfer studies to the in vivo situation. The similar variance in measured and predicted (based on Day 15 length) Day 7 embryo stage indicate low stochastic perturbations for in vivo embryo growth (large stochastic growth effects would generate a significantly larger standard deviation in measured embryo length on Day 15). We also identified that Day 7 embryo stage could be predicted based on the Day 7 gene expression profile (58% overall success rate for classification of 5 embryo stages). Our analysis also associated genes with each developmental stage and demonstrates the high level of temporal regulation of genes that occurs during early embryonic development.


Coculture of mouse IL-3-dependent mast cells with 3T3 fibroblasts stimulates synthesis of globopentaosylceramide (Forssman glycolipid) by fibroblasts and surface expression on both populations.
Mouse IL-3-dependent bone marrow culture-derived mast cells (BMMC) and mouse 3T3 fibroblasts, cultured separately or together, were examined for their cell surface expression and biosynthesis of globopentaosylceramide, a marker of the mouse serosal mast cell. As assessed by flow cytometric analysis, BMMC cultured for up to 7 wk in 50% WEHI 3-conditioned medium containing IL-3 did not bind the B1.1 anti-globopentaosylceramide mAb (six experiments). A total of 10 +/- 4% (mean +/- SD, three experiments) of 3T3 fibroblasts that had reached confluence in medium without IL-3 bound B1.1 antibody and, after an additional approximately 28 days of culture in that medium or in 50% WEHI 3-conditioned medium, 12 +/- 3% (mean +/- SD, five experiments) and 16 +/- 7% (mean +/- SD, three experiments) of the cells, respectively, bound the antibody. After coculture of BMMC and confluent 3T3 fibroblasts for 28 days in 50% WEHI 3-conditioned medium, followed by dispersal and purification of the cells, 92 +/- 18% of the mast cells and 92 +/- 16% (mean +/- SD, seven experiments) of the fibroblasts were B1.1+. Whereas the increase in the expression of the epitope bound by B1.1 antibody on fibroblasts was noted by day 14 of coculture, expression of the epitope on mast cells did not occur until day 21 (three experiments). Biosynthesis of globopentaosylceramide was assessed by intrinsic radiolabeling of each cell population and identification of the extracted neutral glycosphingolipids by TLC and autoradiography. Synthesis of globopentaosylceramide was not detected in extracts of 9 x 10(6) BMMC, 1 x 10(6) confluent 3T3 fibroblasts cultured alone for 28 days, or 9 x 10(6) mast cells purified from 28-day cocultures but was readily detected in extracts of 3 x 10(5) fibroblasts purified from the same cocultures. These findings indicate that BMMC stimulate an increase in the synthesis and expression of globopentaosylceramide on 3T3 fibroblasts and suggest that the subsequent appearance of this neutral glycosphingolipid on the surface of the mast cells is due to its secretion by fibroblasts and adsorption to the mast cell surface. Thus, the interactions between mast cells and fibroblasts during coculture alter the biochemical and Ag phenotypes of both populations.


Novel monorail infusion catheter for volumetric coronary blood flow measurement in humans: in vitro validation.
The aim of this study is to validate a novel monorail infusion catheter for thermodilution-based quantitative coronary flow measurements. Based on the principles of thermodilution, volumetric coronary flow can be determined from the flow rate of a continuous saline infusion, the temperature of saline when it enters the coronary artery, and the temperature of the blood mixed with the saline in the distal part of the coronary artery. In an in vitro set-up of the systemic and coronary circulation at body temperature, coronary flow values were varied from 50-300 ml/min in steps of 50 ml/min. At each coronary flow value, thermodilution-based measurements were performed at infusion rates of 15, 20, and 30 ml/min. Temperatures and pressures were simultaneously measured with a pressure/temperature sensor-tipped guidewire. Agreement of the calculated flow and the measured flow as well as repeatability were assessed. A total of five catheters were tested, with a total of 180 measurements. A strong correlation (ρ=0.976, p<0.0001) and a difference of -6.5±15.5 ml/min were found between measured and calculated flow. The difference between two repeated measures was 0.2%±8.0%. This novel infusion catheter used in combination with a pressure/temperature sensor-tipped guidewire allows accurate and repeatable absolute coronary flow measurements. This opens a window to a better understanding of the coronary microcirculation.


Carlactone-independent seedling morphogenesis in Arabidopsis.
Strigolactone hormones are derived from carotenoids via carlactone, and act through the α/β-hydrolase D14 and the F-box protein D3/MAX2 to repress plant shoot branching. While MAX2 is also necessary for normal seedling development, D14 and the known strigolactone biosynthesis genes are not, raising the question of whether endogenous, canonical strigolactones derived from carlactone have a role in seedling morphogenesis. Here, we report the chemical synthesis of the strigolactone precursor carlactone, and show that it represses Arabidopsis shoot branching and influences leaf morphogenesis via a mechanism that is dependent on the cytochrome P450 MAX1. In contrast, both physiologically active Z-carlactone and the non-physiological E isomer exhibit similar weak activity in seedlings, and predominantly signal through D14 rather than its paralogue KAI2, in a MAX2-dependent but MAX1-independent manner. KAI2 is essential for seedling morphogenesis, and hence this early-stage development employs carlactone-independent morphogens for which karrikins from wildfire smoke are specific surrogates. While the commonly employed synthetic strigolactone GR24 acts non-specifically through both D14 and KAI2, carlactone is a specific effector of strigolactone signalling that acts through MAX1 and D14.


APOBEC3G Is a p53-Dependent Restriction Factor in Respiratory Syncytial Virus Infection of Human Cells Included in the p53/Immune Axis.
Identifying and understanding genetic factors that influence the propagation of the human respiratory syncytial virus (RSV) can lead to health benefits and possibly augment recent vaccine approaches. We previously identified a p53/immune axis in which the tumor suppressor p53 directly regulates the expression of immune system genes, including the seven members of the APOBEC3 family of DNA cytidine deaminases (A3), which are innate immune sentinels against viral infections. Here, we examined the potential p53 and A3 influence in RSV infection, as well as the overall p53-dependent cellular and p53/immune axis responses to infection. Using a paired p53 model system of p53+ and p53- human lung tumor cells, we found that RSV infection activates p53, leading to the altered p53-dependent expression of A3D, A3F, and A3G, along with p53 site-specific binding. Focusing on A3G because of its 10-fold-greater p53 responsiveness to RSV, the overexpression of A3G can reduce RSV viral replication and syncytial formation. We also observed that RSV-infected cells undergo p53-dependent apoptosis. The study was expanded to globally address at the transcriptional level the p53/immune axis response to RSV. Nearly 100 genes can be directly targeted by the p53/immune axis during RSV infection based on our p53BAER analysis (Binding And Expression Resource). Overall, we identify A3G as a potential p53-responsive restriction factor in RSV infection. These findings have significant implications for RSV clinical and therapeutic studies and other p53-influenced viral infections, including using p53 adjuvants to boost the response of A3 genes.


Public Perspectives on Anti-Diabetic Drugs: Exploratory Analysis of Twitter Posts.
Diabetes mellitus is a major global public health issue where self-management is critical to reducing disease burden. Social media has been a powerful tool to understand public perceptions. Public perception of the drugs used for the treatment of diabetes may be useful for orienting interventions to increase adherence. The aim of this study was to explore the public perceptions of anti-diabetic drugs through the analysis of health-related tweets mentioning such medications. This study uses an infoveillance social listening approach to monitor public discourse using Twitter data. We coded 4000 tweets from January 1, 2019 to October 1, 2019 containing key terms related to anti-diabetic drugs by using qualitative content analysis. Tweets were coded for whether they were truly about an anti-diabetic drug and whether they were health-related. Health-related tweets were further coded based on who was tweeting, which anti-diabetic drug was being tweeted about, and the content discussed in the tweet. The main outcome of the analysis was the themes identified by analyzing the content of health-related tweets on anti-diabetic drugs. We identified 1664 health-related tweets on 33 anti-diabetic drugs. A quarter (415/1664) of the tweets were confirmed to have been from people with diabetes, 17.9% (298/1664) from people posting about someone else, and 2.7% (45/1664) from health care professionals. However, the role of the tweeter was unidentifiable in two-thirds of the tweets. We identified 13 themes, with the health consequences of the cost of anti-diabetic drugs being the most extensively discussed, followed by the efficacy and availability. We also identified issues that patients may conceal from health care professionals, such as purchasing medications from unofficial sources. This study uses an infoveillance approach using Twitter data to explore public perceptions related to anti-diabetic drugs. This analysis gives an insight into the real-life issues that an individual faces when taking anti-diabetic drugs, and such findings may be incorporated into health policies to improve compliance and efficacy. This study suggests that there is a fear of not having access to anti-diabetic drugs due to cost or physical availability and highlights the impact of the sacrifices made to access anti-diabetic drugs. Along with screening for diabetes-related health issues, health care professionals should also ask their patients about any non-health-related concerns regarding their anti-diabetic drugs. The positive tweets about dietary changes indicate that people with type 2 diabetes may be more open to self-management than what the health care professionals believe.


Case report and review of clenbuterol cardiac toxicity.
Clenbuterol is an oral β2-agonist utilized as an illicit substance for performance-enhancement or weight loss. We report a case of a 23-year-old male who presented with anxiety and chest tightness after intentional ingestion of 5000 μg of clenbuterol (125 times the recommended adult dose) to lose weight. His electrocardiogram showed sinus tachycardia and diffuse nonspecific repolarization abnormalities with mild inferolateral ST-segment depression. Bloodwork revealed a potassium of 2.0 mmol/L, peak lactate of 9.4 mmol/L, and peak troponin of 5.39 μg/L. A transthoracic echocardiogram was normal except for hyperdynamic left ventricular function. He was treated with intravenous fluids and oral metoprolol. His tachycardia and electrocardiogram abnormalities resolved after 48 h. Clenbuterol has gained notoriety in recent years as a drug of abuse and cases of toxicity will likely continue to increase due to its relative attainability and readily accessible online dosing information. Patients often present with agitation, palpitations, tachycardia, hypokalemia, and hyperglycemia. Treatment is supportive with intravenous fluids, β-blockers, and potassium supplementation. <Learning objective: Clenbuterol, an oral β2-agonist, can be utilized as an illicit substance for performance-enhancement or weight loss. Cardiac toxicity and type II myocardial infarction can occur with clenbuterol overdoses. Associated symptoms include agitation, palpitations, tachycardia, hypokalemia, and hyperglycemia. Treatment is supportive primarily with intravenous fluid, β-blockers, and potassium supplementation.>.


Brain imaging studies of cocaine abuse: implications for medication development.
Contemporary in vivo brain imaging techniques confer the ability to assess brain function and structure noninvasively, and thereby can yield information to help guide the development of new treatments for substance abuse. The advantages and limitations of the major imaging modalities (positron emission tomography [PET], single photon emission computed tomography [SPECT], structural and functional magnetic resonance imaging [MRI, fMRI, respectively]) are discussed with respect to their applicability to research on cocaine abuse. The effects of acute administration of cocaine have been studied using PET and fMRI, with PET manifesting decreases in cerebral glucose metabolism and blood flow, and fMRI revealing regional effects that are correlated temporally with subjective responses. In addition, studies of drug abusers, abstinent from cocaine for various lengths of time, have revealed persistent differences in brain function and structure, especially in the frontal cortex, when compared with parameters in the brains of subjects who do not use illicit drugs of abuse. PET studies also have revealed abnormalities in markers for dopaminergic and opioid systems during withdrawal from cocaine. Moreover, studies of cue-elicited craving for cocaine demonstrate a connection between the response to drug-related stimuli and neural elements of cognition and emotion. The future directions of in vivo brain imaging to identify functional and structural alterations in the brains of cocaine abusers are discussed in relation to the development of medications to treat cocaine dependence.


The effects of anxiety and exercise-induced fatigue on shooting accuracy and cognitive performance in infantry soldiers.
Operational performance in military settings involves physical and mental skills that are generally investigated separately in lab settings, leading to reduced ecological validity. Therefore, we investigated the effects of anxiety and exercise-induced fatigue, separately and in combination, on cognitive and shooting performance of 22 soldiers in a real-world setting. Findings indicated that soldiers' shooting accuracy and decision-making and mathematical skills decreased significantly under anxiety. Whether exercise-induced fatigue was beneficial or detrimental to task performance depended on the task at hand. The increased arousal levels through exercise prevented shooting accuracy from deteriorating in the decision task. In contrast, cognitive performance suffered from the increased arousal: participants more often failed to shoot when being fired at by an opponent and also math performance seemed to decrease. We conclude that anxiety can deteriorate soldier performance and that exercise-induced fatigue may improve or deteriorate performance in combination with anxiety depending on the nature of the task. Soldiers encounter anxiety and exercise-induced fatigue. We investigated to what degree these factors influence soldiers' shooting and cognitive performance. Experimental manipulation of anxiety and exercise during a representative field course indicated decreased performance under anxiety. Exercise prevented shooting accuracy from deteriorating under anxiety, although cognitive performance was negatively affected after exercise.


Predictors of Quality of Life Among Omani Family Caregivers of Patients With Traumatic Brain Injury.
This study aimed to explore the quality of life (QoL) of Omani family caregivers (FCs) of patients with traumatic brain injury (TBI). After acute care, the burden of caring for patients with TBI is mainly shouldered by FCs. This cross-sectional study was conducted at Khoula Hospital, Muscat, Oman. Patient data were collected at the time of discharge and 8 weeks post-discharge between April 2019 and December 2021. The SF-12 General Health Survey and Preparedness for Caregiving scale were used to measure the caregivers' QoL and preparedness, respectively. The Disability Rating and TBI Symptom scales were used to measure the patients' disability and symptoms, respectively. A total of 36 FCs and patients with TBI were recruited. Most caregivers were the parent (41.7%) or child (27.8%) of the patient with TBI. Overall, the caregivers had good physical QoL (PQoL) and mental health QoL (MHQoL) but low caregiving preparedness at the time of discharge. At 8 weeks post-discharge, there were significant improvements in caregiving preparedness (P <0.01) and patient disability (P <0.05) but a depreciation in caregivers' MHQoL (P <0.05), with no change in their PQoL. The caregiver's employment status and the severity of the patient's sleep and mood problems were modifiable predictors of the caregivers' PQoL. Caregiver's preparedness, the patient's inability to live independently, and the severity of the patient's mood and behavioural problems were predictors of caregivers' MHQoL. Caring for patients with TBI negatively impacts the QoL of Omani FCs; this correlates with the physical, emotional, and mental health symptoms of the patient.


Primary superficial vein reflux with competent saphenous trunk.
because reflux in superficial vein tributaries is most often collectively reported with the main saphenous veins, its importance remains largely unrecognised. This study was designed to identify the distribution and extent of non-truncal superficial venous reflux and its association with the signs and symptoms of chronic venous disease (CVD). eighty-four limbs in 62 patients with signs and symptoms of CVD and evidence of reflux on continuous-wave Doppler were subsequently examined with colour-flow duplex imaging. Incompetent superficial vein tributaries were imaged throughout their extent and both ends were identified. Limbs with reflux in the main trunk of the saphenous veins or the deep, perforator or muscular veins, superficial or deep vein thrombosis, injection sclerotherapy, varicose-vein surgery, arterial disease and inflammation of non-venous origin were excluded from the study. The CEAP classification system was used for staging clinical severity of CVD. the prevalence of tributary reflux alone was 9.7% (84/860). Reflux was detected in 171 tributaries. The number of incompetent tributaries ranged from 1 to 5 per limb. Most prevalent were the tributaries to the greater saphenous (111, 65%<0. 0001), followed by those of lesser saphenous (33, 19%) or a combination of both (12, 7%). Incompetent non-saphenous tributaries were uncommon (15, 9%). Among the named tributaries in the lower limb the posterior arch vein was most often incompetent (46, 27%) followed by the anterolateral vein of the thigh (30, 18%), the medial accessory vein (16, 9%) and the anterior arch vein (14, 8%). Reflux in above-the-knee tributaries alone was found in 18 limbs (21%), in below the knee in 23 (28%) and in both sites in 43 (51%). The vast majority of the limbs (71%,p <0.0001) belonged to CVD class 2, 14% in class 3, 9% in class 1 and only 6% in class 4. Class 3 and 4 patients tended to have a longer duration of signs and symptoms, higher number of incompetent tributaries per limb and also a higher prevalence of combined above- and below-knee reflux. these data indicate that reflux confined to superficial tributaries is found throughout the lower limb. Because this reflux is present without greater and lesser saphenous trunk, perforator and deep-vein incompetence or proximal obstruction, it shows that reflux can develop in any vein without an apparent feeding source. Greater saphenous tributaries are affected significantly more often than those of lesser saphenous, while non-saphenous reflux is uncommon. Most limbs have signs and symptoms of CVD class 2 and 15% belong in classes 3 and 4.


Translating the Results of Discrete Choice Experiments into p-/e-/m-Health Decision Support Tools.
The rapidly growing number of health-related Discrete Choice Experiments (DCEs) has not been matched by studies of their impact on decision or policymaking. However, it is widely assumed that this impact has been very limited, despite the potential relevance of the resulting average preferences to group policy development. The main, but at the moment essentially speculative, explanation offered, focuses on the methodological quality of the DCEs and their reporting. An alternative explanation, equally speculative, lies in the research-practice gap created by the conceptualisation of the DCE as a purely research exercise, not supplemented by any attempt to translate the findings into analytic decision support form. This also applies in the clinical decision context, where there are frequent claims that DCE results can assist in an individual's decision making. In the absence of suggestions as to how group results can analytically facilitate preference-sensitive care (and legally informed consent), we propose a generic add-on for DCEs with 'real' options, attributes, and attribute levels. This takes the form of a multi-criteria analysis-based decision support tool. Exemplars, showing how preference-sensitive individualised opinions can be derived from published DCEs for Heavy Menstrual Bleeding and Prostate Cancer Screening, may be consulted online.


WT1 (Wilms' tumor gene 1): biology and cancer immunotherapy.
Wilms' tumor gene WT1 encodes a transcription factor and plays an important role in cell growth and differentiation. The WT1 gene is highly expressed in leukemia and various types of solid tumors, whereas WT1 is a tumor marker convenient for the detection of minimal residual disease of leukemia. The WT1 gene was originally defined as a tumor suppressor gene, but we proposed that it was, on the contrary, an oncogene. Furthermore, the WT1 protein has proven to be a promising tumor-associated antigen, in which many human leukocyte antigen class I- or II-restricted WT1 epitopes have been identified. Clinical trials of WT1-targeted immunotherapy have confirmed its safety and clinical efficacy. WT1-specific cytotoxic T lymphocytes and WT1 antibodies are spontaneously induced in tumor-bearing patients, probably because of high immunogenicity of the WT1 protein. WT1-specific cytotoxic T lymphocytes make a major contribution to the graft-versus-leukemia effect after allogenic stem cell transplantation. When 75 cancer antigens including WT1 were prioritized according to several criteria such as therapeutic function and immunogenicity, WT1 was ranked as the top antigen. These findings suggest that a new era of WT1 immunotherapy is imminent.


Dietary patterns and asthma in the E3N study.
The aim of the present study was to determine dietary patterns and investigate their association with asthma incidence, current asthma and frequent asthma exacerbations. Dietary habits and asthma data were collected from the large E3N study (of French females, mostly teachers). Of the 54,672 females followed-up in 2003, 2,634 reported ever-adulthood asthma, 1,063 reported current asthma, 206 reported frequent asthma attacks (one or more a week), and 628 reported asthma onset between 1993 and 2003. Using principal component analysis, three dietary patterns were identified: the "prudent" pattern (fruits and vegetables); the "Western" pattern (pizza/salty pies, dessert and cured meats); and the "nuts and wine" pattern. Pattern scores were categorised into tertiles, and the incidence and prevalence of asthma was compared between tertiles. After adjustment for confounders, no association of dietary patterns was observed with asthma incidence, ever-asthma or current asthma. The Western pattern was associated with an increased risk of reporting frequent asthma attacks (highest versus lowest tertile odds ratio (OR) 1.79, 95% confidence interval (CI) 1.11-3.73). Increasing scores of the nuts and wine pattern were associated with a decreased risk of reporting frequent asthma attacks (highest versus lowest tertile OR 0.65, 95% CI 0.31-0.96). The results of the present study suggest that overall diet could be involved in frequent asthma exacerbations, one aspect of asthma severity.


Treatment of the Pregnant Patient with Breast Cancer.
Pregnancy-associated breast cancer is defined as invasive breast cancer diagnosed during gestation, within 1 year postpartum, or during lactation. Of particular interest is the treatment of invasive breast cancer during gestation; standard treatment protocols must take into account the health of the fetus. This article reviews the literature and emerging data regarding the treatment of pregnancy-associated breast cancer. Existing staging and treatment practices need slight modification in the setting of pregnancy. The timing of surgery and the administration of cytotoxic chemotherapy must take into account age of gestation, but these modalities are safe in pregnancy.


Metatarsal Stress Fractures.
Metatarsal stress fractures are also called "march fractures" or "marcher's foot." They most commonly occur in the distal second and third metatarsals. The second and third metatarsals receive the majority of stress during ambulation and are less mobile compared with the other metatarsals. The predominant risk factor is excessive overuse with repetitive motions with little recovery time such as marching activities in the military. Any element that has a detrimental effect on bone density can predispose an individual to stress fractures. There is often a delay of 2 or more weeks from the onset of symptoms until visible fractures are able to be seen on radiographs. Initial treatment strategies involve rest, ice, nonweight bearing, and avoidance of exercise to prevent fracture displacement, nonunion, and other complications. Orthopedic referral will guide definitive care.


Systematic review: body composition in adults with inflammatory bowel disease.
There is a paucity of data on body composition in patients with inflammatory bowel disease (IBD). Alterations of fat and muscle may affect bone health, muscle performance, quality of life (QoL) and overall morbidity. To systematically review the literature on body composition in adults with IBD, and to discuss potential contributory factors and associations. A systematic search was performed in July 2012 of OVID SP MEDLINE, OVID EMBASE and National Library of Medicine's PubMed Central Medline (Limitations: English, humans, from 1992). A total of 19 articles comparing body composition in patients with IBD with healthy age- and sex-matched control populations were included in the primary analysis. A total of 631 patients with Crohn's disease (CD) and 295 with ulcerative colitis (UC), mean age 37.1 (s.d. ± 9.2) years; 485 (52%) female, were reported upon. Data were heterogeneous and methodology varied. Compared with controls, a statistically significant reduction in body mass index (BMI) was reported in 37% of CD and 20% of UC patients; reduced fat-free mass in 28% CD and 13% UC patients, and reduced fat mass in 31% CD and 13% UC patients. There was no consistent association between body composition and disease activity, duration, extent or therapies. BMI did not accurately predict body composition. Current data, although heterogeneous, suggest that many patients with IBD are affected by aberrations in fat and lean mass, which may not be detected during routine clinical assessment. The prevalence and impact of altered body composition amongst this population warrant further investigation.


Strontium isotopic aspects of Paranthropus robustus teeth; implications for habitat, residence, and growth.
The strontium isotopic ratio 87Sr/86Sr has been studied in the Sterkfontein Valley of South Africa to infer both habitat usage and residence for a number of early hominins. This paper examines the existing 87Sr/86Sr data collectively derived from three studies of Paranthropus robustus teeth with the aim of exploring whether the dataset as a whole may provide deeper insight into habitat, mobility, and growth for this species. 87Sr/86Sr from seven Swartkrans Member I third molars varies in a well defined narrow range, and while some canines were consistent with this range, a number of P. robustus canines and first and second molars were not, and therefore represent individuals who had arrived from other localities. A first and third molar 87Sr/86Sr was found to differ in TM1517c, the holotype P. robustus specimen from Kromdraai, suggesting this individual had moved to the locality sometime after the first molar and before the third molar had completely mineralized. While early forming teeth vary widely, the relatively low variation and absence of exogenous 87Sr/86Sr in third molars suggest that these teeth mineralized relatively late when compared to life history events bearing on higher primate residence patterns. The implications for further study of habitat, residence, and growth are discussed.


Transduction pathways regulating the trophic effects of Saccharomyces boulardii in rat intestinal mucosa.
Saccharomyces boulardii is a probiotic yeast that is widely prescribed in lyophilized form; it determines several effects in human and rat small intestine including endoluminal secretion of enzymes and of polyamines, stimulation of microvillous enzymes, of sIgA, increased production of the receptor for polymeric immunoglobulins by crypt cells, and enhanced d-glucose uptake. The objective of this study was to determine the pathway(s) by which these effects generated by the yeast are transduced into mucosal cells. Litters of six growing Wistar rats each were treated with S. boulardii (50 microg/gram body weight) or with saline between days 30 and 34 postpartum. For each animal, the cytosol was prepared from the whole mucosa after the fat cake was discarded. Several known intestinal substrates were immunoprecipitated and immunoblotted using specific antibodies recognizing the non-, mono-, or diphosphorylated forms of these substrates. The signals were detected using Echochemiluminoscence (ECL) and were measured by optodensitometry. Treatment with S. boulardii markedly enhanced the RAS-GAP-RAF-ERK(1,2) pathway with participation of growth receptor bound 2 protein, SHC, SOS, and CRKII. Unit p85alpha of phosphatidylinositol 3 kinase, tested in its phosphorylated form, was also enhanced by the probiotic compared to control samples. In rats treated with an inhibitor of RAF-1 and of ERK(1,2) (PD098059) the expression of mucosal disaccharidases was inhibited by about 50%. The probiotic S. boulardii generates in vivo mitogen and metabolic signals that are transduced into intestinal mucosal cells, downstream from the apical membrane to the nuclei, using recruitment substrates and serine, threonine, or tyrosine kinases.