Interleukin-35 Expression in Non-Small Cell Lung Cancer is Associated with Tumor Progression.
Lung cancer continues to be the leading cause of cancer related deaths worldwide due to its high incidence, malignant behavior and lack of major advancements in treatment strategy. The occurrence and development of lung cancer is closely related to inflammation. Thus, we conducted the present study to investigate the effects of IL-35 (Interleukin 35), a newly identified anti-inflammatory factor, on non-small cell lung cancer (NSCLC), which accounts for about 85% of all lung cancers. We first evaluated the IL-35 expression in 384 pairs of NSCLC samples and their adjacent normal mucosa by realtime PCR, ELISA (Enzyme-linked immunoassay) and tissue microarrays. Then the role of IL-35 on patient survival rates, cancer progression and their sensitivity to chemotherapy drugs were assessed. IL-35 was barely expressed in the NSCLC tissues but highly expressed in the adjacent normal tissues. The down-regulation of IL-35 was significantly correlated with the results of American Joint Committee on Cancer stage, differentiation and it was also shown to be an independent prognostic indicator of disease-free survival and overall survival for patients with NSCLC. Overexpression of IL-35 in NSCLC cells suppressed cell migration, invasion, proliferation, colony formation through suppressing β-catenin. IL-35 inhibited NSCLC formation in the mice model and sensitize the cancer cells to chemotherapy drugs. Our results showed that IL-35 plays an inhibitory role in NSCLC development and function as a novel prognostic indicator and a potential therapeutic target.


Column selection for comprehensive multidimensional ion chromatography.
This paper discusses the selection of ion chromatography (IC) columns for use in comprehensive multidimensional ion chromatography (IC x IC). First, a single number was determined for a wide range of anions (one number for each anion) using the linear solvent strength model. These numbers were then used to compare the column selectivity characteristics for five different columns. Principal component analysis was used to illustrate selectivity differences between columns. Dionex AS16 and AS20 columns were selected for use in the development of an IC x IC method for the separation of ten anions. To achieve the required speed of analysis in both the first and second separation dimensions, custom column lengths were packed in-house. The use of an eluent suppressor between the first and second columns permits a relatively low flow ratio regime of only <1:20 in the first and second dimensions, respectively, which reduces dilution effects common in comprehensive multidimensional LC. Selection of the second dimension eluent conditions was aided by the development of a spreadsheet based on the linear solvent strength model.


Ethics of antibiotic allergy.
Antibiotic allergies are commonly reported among patients, but most do not experience reactions on rechallenge with the same agents. These reported allergies complicate management of infections in patients labelled as having penicillin allergy, including serious infections where penicillin-based antibiotics are the first-line (most effective and least toxic) treatment option. Allergy labels are rarely questioned in clinical practice, with many clinicians opting for inferior second-line antibiotics to avoid a perceived risk of allergy. Reported allergies thereby can have significant impacts on patients and public health, and present major ethical challenges. Antibiotic allergy testing has been described as a strategy to circumvent this dilemma, but it carries limitations that often make it less feasible in patients with acute infections or in community settings that lack access to allergy testing. This article provides an empirically informed ethical analysis of key considerations in this clinical dilemma, using Staphylococcus aureus bacteraemia in patients with penicillin allergies as a case study. We argue that prescribing first-line penicillin-based antibiotics to patients with reported allergies may often present a more favourable ratio of benefits to risks, and may therefore be more ethically appropriate than using second-line drugs. We recommend changes to policy-making, clinical research and medical education, in order to promote more ethically acceptable responses to antibiotic allergies than the status quo.


A case report of a fetus with mosaic autosomal variegated aneuploidies and literature review.
Mosaic variegated aneuploidy (MVA) is a recessive condition characterized by mosaic aneuploidies, predominantly trisomies and monosomies, involving multiple chromosomes and tissues. The phenotype of MVA syndrome includes severe microcephaly and growth deficiency, central nervous system anomalies, mental retardation, mild physical anomalies, and predisposition to cancer. We report a case of true fetal mosaicism for variegated aneuploidies detected in amniotic fluid cells. A 33-year-old primigravida woman at 5 weeks 1 day of gestation was referred to our tertiary hospital because of a high-risk pregnancy associated with IgA nephropathy. In a quadruple screening test performed at the 15(th) week of gestation, alpha fetoprotein was 73.4 IU/mL (2.792 MoM), suggesting that she was at high risk of neural tube defect. Following amniocentesis performed at the 17 weeks' gestation, chromosome examination of amniocyte culture showed premature chromatic separation in 63% of the metaphases (58/92) and a high frequency of gain and loss of chromosomes. Repeat amniocentesis at 21 weeks' gestation consistently showed the presence of multiple mosaic autosomal variegated aneuploidies. Ultrasonography at 21 weeks' gestation revealed relatively small head circumference for gestational age (<3%) and vermis defect, suggesting that the fetus would have microcephaly and Dandy-Walker malformation. Cytogenetic analysis with peripheral blood of the parents showed normal karyotype. In summary, we hereby report the cytogenetic analysis and prenatal findings of MVA.


Brown adipose tissue regulates glucose homeostasis and insulin sensitivity.
Brown adipose tissue (BAT) is known to function in the dissipation of chemical energy in response to cold or excess feeding, and also has the capacity to modulate energy balance. To test the hypothesis that BAT is fundamental to the regulation of glucose homeostasis, we transplanted BAT from male donor mice into the visceral cavity of age- and sex-matched recipient mice. By 8-12 weeks following transplantation, recipient mice had improved glucose tolerance, increased insulin sensitivity, lower body weight, decreased fat mass, and a complete reversal of high-fat diet-induced insulin resistance. Increasing the quantity of BAT transplanted into recipient mice further improved the metabolic effects of transplantation. BAT transplantation increased insulin-stimulated glucose uptake in vivo into endogenous BAT, white adipose tissue (WAT), and heart muscle but, surprisingly, not skeletal muscle. The improved metabolic profile was lost when the BAT used for transplantation was obtained from Il6-knockout mice, demonstrating that BAT-derived IL-6 is required for the profound effects of BAT transplantation on glucose homeostasis and insulin sensitivity. These findings reveal a previously under-appreciated role for BAT in glucose metabolism.


Hepatitis B and hepatitis C virus replication upregulates serine protease inhibitor Kazal, resulting in cellular resistance to serine protease-dependent apoptosis.
Hepatitis B and C viruses (HBV and HCV, respectively) are different and distinct viruses, but there are striking similarities in their disease potential. Infection by either virus can cause chronic hepatitis, liver cirrhosis, and ultimately, liver cancer, despite the fact that no pathogenetic mechanisms are known which are shared by the two viruses. Our recent studies have suggested that replication of either of these viruses upregulates a cellular protein called serine protease inhibitor Kazal (SPIK). Furthermore, the data have shown that cells containing HBV and HCV are more resistant to serine protease-dependent apoptotic death. Since our previous studies have shown that SPIK is an inhibitor of serine protease-dependent apoptosis, it is hypothesized that the upregulation of SPIK caused by HBV and HCV replication leads to cell resistance to apoptosis. The evasion of apoptotic death by infected cells results in persistent viral replication and constant liver inflammation, which leads to gradual accumulation of genetic changes and eventual development of cancer. These findings suggest a possibility by which HBV and HCV, two very different viruses, can share a common mechanism in provoking liver disease and cancer.


Organochlorine pesticides (OCPs) in South Asian region: a review.
Organochlorine pesticides (OCPs) are semi volatile organic compounds of global concern. During the last decades, their distribution, sources, transformation, toxicity and accumulation in the terrestrial and aquatic ecosystems have gained significant attention. Many of these chemicals are characterized by long range atmospheric transport potential, and their occurrence in remote areas is attributed to transport of chemicals from places where OCPs are still in use. The South Asia region is a place where primarily emissions are still taking place and thus it is important to assess the status of OCPs pollution. This document provides the historical overview and country specific environmental legislation of OCPs from the South Asian region in the context of their illegal use and storage for extended periods and still until to date. In addition, the current review discusses the existing knowledge on the levels and distribution of OCPs in different environmental compartments of South Asian region. Data on OCPs also highlights the risk assessment of these organic contaminants in the regional environment and spans the long range atmospheric transport phenomena based on Himalayas and Northern mountainous glaciers. Paradoxically in the scientific literature sources, distribution and transport of these organic pollutants in South Asian region are very limited compared to the rest of the world.


Enzyme-linked immunosorbent assay (ELISA) for the anthropogenic marker isolithocholic acid in water.
Bile acids are promising chemical markers to assess the pollution of water samples with fecal material. This study describes the optimization and validation of a direct competitive enzyme-linked immunosorbent assay for the bile acid isolithocholic acid (ILA). The quantification range of the optimized assay was between 0.09 and 15 μg/L. The assay was applied to environmental water samples. Most studies until now were focused on bile acid fractions in the particulate phase of water samples. In order to avoid tedious sample preparation, we undertook to evaluate the dynamics and significance of ILA levels in the aqueous phase. Very low concentrations in tap and surface water samples made a pre-concentration step necessary for this matrix as well as for wastewater treatment plant (WWTP) effluent. Mean recoveries for spiked water samples were between 97% and 109% for tap water and WWTP influent samples and between 102% and 136% for WWTP effluent samples. 90th percentiles of intra-plate and inter-plate coefficients of variation were below 10% for influents and below 20% for effluents and surface water. ILA concentrations were quantified in the range of 33-72 μg/L in influent, 21-49 ng/L in effluent and 18-48 ng/L in surface water samples. During wastewater treatment the ILA levels were reduced by more than 99%. ILA concentrations of influents determined by ELISA and LC-MS/MS were in good agreement. However, findings in LC-ELISA experiments suggest that the true ILA levels in concentrated samples are lower due to interfering effects of matrix compounds and/or cross-reactants. Yet, the ELISA will be a valuable tool for the performance check and comparison of WWTPs and the localization of fecal matter input into surface waters.


Chronic selective glycogen synthase kinase-3 inhibition enhances glucose disposal and muscle insulin action in prediabetic obese Zucker rats.
Increasing evidence supports a negative role of glycogen synthase kinase-3 (GSK-3) in regulation of skeletal muscle glucose transport. We assessed the effects of chronic treatment of insulin-resistant, prediabetic obese Zucker (fa/fa) rats with a highly selective GSK-3 inhibitor (CT118637) on glucose tolerance, whole body insulin sensitivity, plasma lipids, skeletal muscle insulin signaling, and in vitro skeletal muscle glucose transport activity. Obese Zucker rats were treated with either vehicle or CT118637 (30 mg/kg body wt) twice per day for 10 days. Fasting plasma insulin and free fatty acid levels were reduced by 14 and 23% (P < 0.05), respectively, in GSK-3 inhibitor-treated animals compared with vehicle-treated controls. The glucose response during an oral glucose tolerance test was reduced by 18% (P < 0.05), and whole body insulin sensitivity was increased by 28% (P < 0.05). In vivo insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation (50%) and IRS-1-associated phosphatidylinositol-3' kinase (79%) relative to fasting plasma insulin levels were significantly elevated (P < 0.05) in plantaris muscles of GSK-3 inhibitor-treated animals. Whereas basal glucose transport in isolated soleus and epitrochlearis muscles was unaffected by chronic GSK-3 treatments, insulin stimulation of glucose transport above basal was significantly enhanced (32-60%, P < 0.05). In summary, chronic treatment of insulin-resistant, prediabetic obese Zucker rats with a specific GSK-3 inhibitor enhances oral glucose tolerance and whole body insulin sensitivity and is associated with an amelioration of dyslipidemia and an improvement in IRS-1-dependent insulin signaling in skeletal muscle. These results provide further evidence that selective targeting of GSK-3 in muscle may be an effective intervention for the treatment of obesity-associated insulin resistance.


BOD1 Is Required for Cognitive Function in Humans and Drosophila.
Here we report a stop-mutation in the BOD1 (Biorientation Defective 1) gene, which co-segregates with intellectual disability in a large consanguineous family, where individuals that are homozygous for the mutation have no detectable BOD1 mRNA or protein. The BOD1 protein is required for proper chromosome segregation, regulating phosphorylation of PLK1 substrates by modulating Protein Phosphatase 2A (PP2A) activity during mitosis. We report that fibroblast cell lines derived from homozygous BOD1 mutation carriers show aberrant localisation of the cell cycle kinase PLK1 and its phosphatase PP2A at mitotic kinetochores. However, in contrast to the mitotic arrest observed in BOD1-siRNA treated HeLa cells, patient-derived cells progressed through mitosis with no apparent segregation defects but at an accelerated rate compared to controls. The relatively normal cell cycle progression observed in cultured cells is in line with the absence of gross structural brain abnormalities in the affected individuals. Moreover, we found that in normal adult brain tissues BOD1 expression is maintained at considerable levels, in contrast to PLK1 expression, and provide evidence for synaptic localization of Bod1 in murine neurons. These observations suggest that BOD1 plays a cell cycle-independent role in the nervous system. To address this possibility, we established two Drosophila models, where neuron-specific knockdown of BOD1 caused pronounced learning deficits and significant abnormalities in synapse morphology. Together our results reveal novel postmitotic functions of BOD1 as well as pathogenic mechanisms that strongly support a causative role of BOD1 deficiency in the aetiology of intellectual disability. Moreover, by demonstrating its requirement for cognitive function in humans and Drosophila we provide evidence for a conserved role of BOD1 in the development and maintenance of cognitive features.


Agalma: an automated phylogenomics workflow.
In the past decade, transcriptome data have become an important component of many phylogenetic studies. They are a cost-effective source of protein-coding gene sequences, and have helped projects grow from a few genes to hundreds or thousands of genes. Phylogenetic studies now regularly include genes from newly sequenced transcriptomes, as well as publicly available transcriptomes and genomes. Implementing such a phylogenomic study, however, is computationally intensive, requires the coordinated use of many complex software tools, and includes multiple steps for which no published tools exist. Phylogenomic studies have therefore been manual or semiautomated. In addition to taking considerable user time, this makes phylogenomic analyses difficult to reproduce, compare, and extend. In addition, methodological improvements made in the context of one study often cannot be easily applied and evaluated in the context of other studies. We present Agalma, an automated tool that constructs matrices for phylogenomic analyses. The user provides raw Illumina transcriptome data, and Agalma produces annotated assemblies, aligned gene sequence matrices, a preliminary phylogeny, and detailed diagnostics that allow the investigator to make extensive assessments of intermediate analysis steps and the final results. Sequences from other sources, such as externally assembled genomes and transcriptomes, can also be incorporated in the analyses. Agalma is built on the BioLite bioinformatics framework, which tracks provenance, profiles processor and memory use, records diagnostics, manages metadata, installs dependencies, logs version numbers and calls to external programs, and enables rich HTML reports for all stages of the analysis. Agalma includes a small test data set and a built-in test analysis of these data. In addition to describing Agalma, we here present a sample analysis of a larger seven-taxon data set. Agalma is available for download at https://bitbucket.org/caseywdunn/agalma. Agalma allows complex phylogenomic analyses to be implemented and described unambiguously as a series of high-level commands. This will enable phylogenomic studies to be readily reproduced, modified, and extended. Agalma also facilitates methods development by providing a complete modular workflow, bundled with test data, that will allow further optimization of each step in the context of a full phylogenomic analysis.


Visual context and practice change the distribution of attention in touch.
It has often been shown that people experience difficulty restricting their tactile attention to one hand when both hands are touched simultaneously, especially if the hands are in close spatial proximity. Taken in conjunction with a growing body of evidence that the sensations arising from stimulation of the skin are referred to locations in external space rather than their positions on the body, these tactile attention findings provide evidence for the spatial distribution of attention in touch. However, the effect of spatial distance between the arms on the distribution of attention has been shown in only a limited number of studies and in single testing sessions. In a set of experiments we examined the stability of the arm separation effect by measuring performance over two sessions and with varying visual information about arm location. Tactile selection was more efficient when the stimulated hands were placed far apart compared with when they were adjacent but the effect was short lived (session 1 only) and only occurred when participants were tested with their eyes open. These findings suggest that somatosensory attention is a flexible, dynamic process, based on coordinates that can vary over time.


The carotid bifurcation plaque: pathologic findings associated with cerebral ischemia.
Embolization from or decreased flow through cervical carotid and vertebral arteries causes ischemic stroke syndromes. Specific pathologic findings were studied in 50 symptomatic patients who underwent 69 carotid endarterectomies. Detailed analyses of their carotid plaques included correlations between photographs of gross specimens, microscopic findings, angiograms, preoperative symptoms and long-term postoperative follow up. Carotid plaques were primarily fibrous with significant (greater than 70%) stenoses encountered in 70% of the arteries. Stenoses were due to simple fibrous thickening in only 20%; the remainder due to intraplaque hemorrhage, atheromatous debris and, least often, luminal thrombus with or without ulceration. Intramural hemorrhage was frequent in plaques associated with focal neurologic symptoms and may have preceded localized collections of atheromatous debris. Ulceration occurred in 1/3 of all plaques, symptomatic or not. It is concluded that the carotid plaques start as fibrointimal thickening evolving to symptomatic stages by the occurrence of one or more of a number of pathologic changes, intraplaque hemorrhage being prominent. A single rational therapeutic regimen seems impossible until patients can be classified according to their pathologic changes diagnosed non-invasively.


Randomised placebo-controlled trial of outpatient (at home) cervical ripening with isosorbide mononitrate (IMN) prior to induction of labour--clinical trial with analyses of efficacy and acceptability. The IMOP study.
To determine whether isosorbide mononitrate (IMN), self-administered vaginally by women at home, improves the process of induction of labour. Randomised double blind placebo-controlled trial. Large UK maternity hospital. Nulliparous women with a singleton pregnancy, cephalic presentation > or = 37 weeks gestation, requiring cervical ripening prior to induction of labour. IMN (n = 177) or placebo (n = 173) self-administered vaginally at home at 48, 32 and 16 hours prior to the scheduled time of admission for induction. Admission to delivery interval and women's experience of induction of labour. IMN did not shorten the admission to delivery interval as compared with placebo [mean difference of -1.6 hours (95% CI -5.1,1.9, P = 0.37)], despite being more effective than placebo in inducing a change in Bishop score [mean difference of 0.65 (95% CI 0.14,1.17, P = 0.013)]. While both groups found the overall experience of home treatment to be positive, (mean score of 3.8/10 +/- 2.3/10 for the IMN group, where 1 = extremely good and 10 = not at all good) women in the placebo group found it marginally more positive than those in the IMN group (just over half a unit on a 10-point scale, P = 0.043). There were no differences between the groups in the pain or anxiety experienced or willingness to take the treatment in a subsequent pregnancy. IMN self-administered vaginally at home does not shorten admission to delivery interval despite a significant effect on cervical ripeness assessed using the Bishop score. However, women report positive views on cervical ripening at home, and the setting deserves further investigation.


Distribution, Cultivar Susceptibility, and Epidemiology of Apium virus Y on Celery in Coastal California.
Apium virus Y (ApVY) is a potyvirus that was recently found to cause crop loss to celery (Apium graveolens) in California. Symptoms on leaves exhibit varying forms of chlorosis and necrosis. Depending on the cultivar, celery petioles could also exhibit extensive necrotic, sunken, elongated lesions. Severely affected plants were unmarketable. Disease incidence surveys found that a susceptible celery (cv. 414) showed 55% (2007) and 71% (2008) disease. Because it was noted that the Apiaceae weed poison hemlock (Conium maculatum) was present in almost all areas where ApVY affected celery, a 4-year survey collected overwintered hemlock from six coastal county regions and tested composite samples for ApVY using reverse transcription-polymerase chain reaction (RT-PCR) and ApVY-specific primers. These plants were consistently positive for ApVY. Seeds collected from these plants were also positive when tested with the same RTPCR method. However, when ApVY-positive hemlock seeds were germinated and the resulting seedlings tested, all results were negative. The failure of ApVY to be transmitted from hemlock seeds to seedlings was further documented by collecting newly germinated hemlock seedlings from the field and testing them with RT-PCR. All such seedlings were negative for ApVY even though large, adjacent, overwintered hemlock plants tested positive. Two crops of celery seed were produced from ApVY-positive mother plants; celery seed from these infected plants likewise tested positive for ApVY, but seedlings grown from the seed lots were negative for ApVY. Twenty-one celery and celeriac cultivars were inoculated with ApVY using viruliferous aphids, planted in a replicated field trial, and then grown to maturity. Seven cultivars remained symptomless, tested negative for ApVY, and showed signs of possible resistance. The epidemiology of disease caused by ApVY in California evidently involves poison hemlock as a common overwintering host with subsequent vectoring of the virus from hemlock to celery via aphids. ApVY was not seedborne in this weed host or in celery in our experiments. Our data suggest that growers can manage this disease by controlling poison hemlock weed populations and by planting celery cultivars that are not susceptible to ApVY.


Perinatal Exposure to Traffic-Related Air Pollution and Atopy at 1 Year of Age in a Multi-Center Canadian Birth Cohort Study.
The role of traffic-related air pollution (TRAP) exposure in the development of allergic sensitization in children is unclear, and few birth cohort studies have incorporated spatiotemporal exposure assessment. We aimed to examine the association between TRAP and atopy in 1-year-old children from an ongoing national birth cohort study in four Canadian cities. We identified 2,477 children of approximately 1 year of age with assessment of atopy for inhalant (Alternaria, Der p, Der f, cat, dog, cockroach) and food-related (milk, eggs, peanuts, soy) allergens. Exposure to nitrogen dioxide (NO2) was estimated from city-specific land use regression models accounting for residential mobility and temporal variability in ambient concentrations. We used mixed models to examine associations between atopy and exposure during pregnancy and the first year of life, including adjustment for covariates (maternal atopy, socioeconomic status, pets, mold, nutrition). We also conducted analyses stratified by time-location patterns, daycare attendance, and modeled home ventilation. Following spatiotemporal adjustment, TRAP exposure after birth increased the risk for development of atopy to any allergens [adjusted odds ratio (aOR) per 10 μg/m3 NO2 = 1.16; 95% CI: 1.00, 1.41], but not during pregnancy (aOR = 1.02; 95% CI: 0.86, 1.22). This association was stronger among children not attending daycare (aOR = 1.61; 95% CI: 1.28, 2.01) compared with daycare attendees (aOR = 1.05; 95% CI: 0.81, 1.28). Trends to increased risk were also found for food (aOR = 1.17; 95% CI: 0.95, 1.47) and inhalant allergens (aOR = 1.28; 95% CI: 0.93, 1.76). Using refined exposure estimates that incorporated temporal variability and residential mobility, we found that traffic-related air pollution during the first year of life was associated with atopy. Sbihi H, Allen RW, Becker A, Brook JR, Mandhane P, Scott JA, Sears MR, Subbarao P, Takaro TK, Turvey SE, Brauer M. 2015. Perinatal exposure to traffic-related air pollution and atopy at 1 year of age in a multi-center Canadian birth cohort study. Environ Health Perspect 123:902-908; http://dx.doi.org/10.1289/ehp.1408700.


Spatial correlations and degree of polarization in homogeneous electromagnetic fields.
We consider stationary electromagnetic fields modeled as superpositions of unpolarized and angularly uncorrelated plane waves and show that in anisotropic case the electric cross-spectral tensor is proportional to the imaginary part of the Green tensor. This is as for blackbody radiation, but here the field need not be in thermal equilibrium. We also evaluate the degree of polarization for a homogeneous but nonisotropic field for which the plane waves propagate within a cone of angles. The results are compared with the known polarization properties of blackbody radiation.


Feasibility of Establishing a Comprehensive Yoga Program and its Dose-Effect Relationship on Cardiovascular Risk Factors and Wellness Parameters: A Pilot Study.
We sought to study the feasibility of establishing a comprehensive, mostly self-directed yoga program in a hospital and its dose-effect relationship on cardiovascular risk factors and quality of life (QoL) measures over six months. Yoga-based techniques (Advanced Yoga Practices; AYP; advancedyogapractices.com) were taught in 12 biweekly group sessions and self-directed practice at home was emphasized. Cardiovascular risk factors were elucidated by interview and review of medical history. Quality of life (QoL) outcomes included the SF-36, the Cohen Perceived Stress Scale (CPSS), and the Hospital Anxiety and Depression Scale (HADS). Risk factors and QoL measures were compared in participants at baseline and six months, as well as between those practicing ≥ 7 times versus < 7 times per week. A total of 22 individuals (19 women, mean age 59 ± 8.7 years) completed the study. At six months, changes were noted in the Mental Component Scale (MCS) of the SF-36 (p=0.0004) and the CPSS (p = 0.022). A greater improvement in CPSS was noted in those practicing ≥ 7 times versus < 7 times a week (p=0.045). No changes were noted in cardiovascular risk factors. The prescription of a self-directed yoga program was feasible in a hospital setting and resulted in improvement in QoL measures at six months. Practicing more than seven times per week correlated with greater improvement in the perception of stress. Thus, at least a once-daily dose of AYP techniques for a significant improvement in perceived stress is an appropriate dose to employ and study in hospital settings.


Mithramycin A inhibits DNA methyltransferase and metastasis potential of lung cancer cells.
Abnormal CpG island hypermethylation of multiple tumor-suppressor genes (TSGs) can lead to the initiation and progression of human cancer. The cytosine of the CpG island on the promoter region is methylated by 5'-cytosine-methyltransferases (DNMTs). Pharmacologic inhibitors of CpG island methylation provide a rational approach to reactivate the TSGs in tumor cells and to restore the critical cellular pathways in cancer cells. Mithramycin A (MMA) is known to be a GC- and CG-rich DNA-binding agent. We sought to determine whether MMA could inhibit CpG island methylation and DNMT expression in lung cancer cells. We found that MMA reduced the CpG island methylation of antimetastasis TSGs, including SLIT2 and TIMP-3 genes, and was associated with the prevention of metastasis. When highly metastatic CL1-5 lung cancer cells were treated with low doses (10 nmol/l) of MMA for 14 days, they reexpressed mRNA levels for these genes. MMA also inhibited the invasion phenotypes of CL1-5 cells as indicated by its inhibition of cancer cell migration using wound-healing and transwell assays. Molecular docking of MMA onto the DNMT1 catalytic domain revealed that MMA might interact with the catalytic pocket of DNMT1. Western blots showed that DNMT1 protein levels were depleted after MMA. These data support the idea that MMA has demethylation and antimetastasis effects on lung cancer cells. This mechanism might be mediated by the interaction of MMA and DNMT1, leading to the depletion of the DNMT1 protein and the reversal of the metastasis phenotype in lung cancer cells.


[Poliomyelitis vaccine contaminated with SV40 and prior exposure to asbestos: cognitive study in a group of car repair workers].
Pleural and pulmonary malignancies are usually associated with previous asbestos exposure and the presence of simian virus 40 (SV40) has been detected in these neoplasms. Our study aimed to investigate the health situation among mechanics servicing buses, ex-exposed to asbestos, who received polio vaccines contaminated by SV 40. We conducted a descriptive study and so we recruited, on the basis of birth date and duration in the current job, 39 mechanics, born since 1950 until 1965, with length of service above 15 years. Of all subjects a clinical examination, a lung function test and a chest radiograph were obtained. More than 30% of sample showed pharynx and larynx clinical alterations and radiological signs of previous exposure to asbestos. We didn't find pleural or pulmonary malignancies; besides 4 doubtful neoplasms required further investigations. Although exiguity of sample, these findings provide a lack of mesothelioma and lung cancer among mechanics, previously exposed to asbestos and infected by SV40.


Phase II study of fluorouracil and its modulation in advanced colorectal cancer: a Southwest Oncology Group study.
A variety of fluorinated pyrimidine-based regimens for the treatment of disseminated colorectal cancer have been presented in the medical literature. The Southwest Oncology Group designed a screening trial of seven regimens of fluorouracil (5-FU) to assess efficacy and toxicity afforded by biochemical modulation or schedule variations. Six hundred twenty patients were entered into this trial between August 1989 and January 1993. Eligible patients were classified as having recurrent or disseminated disease that was measurable or nonmeasurable. All eligible patients were evaluated for toxicity and survival; patients with measurable disease were evaluated for response according to standard criteria. No regimen achieved a higher response rate than single-agent bolus 5-FU. Eighty-four percent of patients have been monitored until death. The median survival time for the entire cohort is 14 months. Survival hazards ratios showed a positive trend in favor of the unmodulated infusion regimens, while high-grade toxicities occurred more frequently in the 5-FU bolus arms. The major high-grade toxicities were granulocytopenia and diarrhea. In this trial, no regimen provided substantial improvement relative to 5-FU bolus or single-agent therapy for either response or survival in the treatment of disseminated colorectal cancer. The single-agent infusion regimens demonstrated the most encouraging results with a favorable toxicity profile and a 2-month longer survival duration than 5-FU bolus therapy.


Diabetic macular edema: changing treatment paradigms.
To review the current management and recent changes in treatment paradigm for diabetic macular edema (DME). During the review period (1 year), several prospective studies analyzed the beneficial effect of anti-vascular endothelial growth factor agents in the management of DME. An exploratory analysis concluded that intravitreal ranibizumab appears to be associated with a reduced risk of diabetic retinopathy worsening. A randomized, controlled, multicenter, double-masked, parallel-group, 12-month trial to evaluate a dexamethasone intravitreal implant (DEX implant) combined with laser photocoagulation compared with laser alone for treatment of DME concluded that there was no significant between-group difference at month 12. A multicenter, prospective, observational study found that in eyes with diabetic retinopathy without concurrent central-involved DME, presence of noncentral-involved DME immediately prior to cataract surgery or history of DME treatment may increase the risk of developing central-involved macular edema after cataract extraction. Another randomized trial to evaluate whether intravitreal ranibizumab injection at cataract surgery prevents postoperative DME concluded that intravitreal ranibizumab injection at cataract surgery may prevent the postoperative worsening of macular edema. The results of clinical trials have shown the superiority of some of these anti-vascular endothelial growth factor agents to laser therapy. However, with the availability of several of these newer agents, it may be difficult to individualize treatment options, especially if DME patients respond differently to various therapies.


Exaggerated status of "novel" and "pathogenic" mtDNA sequence variants due to inadequate database searches.
Given its relative ease, screening the entire mitochondrial DNA (mtDNA) for heteroplasmic or novel homoplasmic mutations has become part of the routine diagnostic workup for the molecular geneticist confronted with a disease case exhibiting clinical and biochemical features of mitochondrial dysfunction. "Novelty" of a given mtDNA variant is most often equated with nonregistration in the extensive MITOMAP database (www.mitomap.org). This practice has led to a number of spurious findings and wrong conclusions concerning the pathogenic status of specific mtDNA mutations, especially in the absence of proper evaluation and pathogenicity scoring. We demonstrate by way of real cases targeting the mt-tRNA(Cys) (MT-TC) gene and a stretch within the MT-ND3 gene, that a straightforward Google search can identify twice as many previously observed mutations than any MITOMAP query could achieve. Further, we reassess the recent rediscovery of m.15287T>C by listing all known occurrences and, where possible, providing the haplogroup context, shedding new light on the potential pathogenicity status of m.15287T>C.


Heteromeric channel formation and Ca(2+)-free media reduce the toxic effect of the weaver Kir 3.2 allele.
Weaver mice have a severe hypoplasia of the cerebellum with an almost complete loss of the midline granule cells. Recent genetic studies of weaver mice have identified a mutation resulting in an amino acid substitution (G156S) in the pore of the inwardly rectifying potassium channel subunit Kir 3.2. When expressed in Xenopus oocytes the weaver mutation alters channel selectivity from a potassium-selective to a nonspecific cation-selective pore. In this study we confirm by cell-attached patch-clamp recording that the mutation produces a non-selective cation channel. We also demonstrate that the cell death induced by weaver expression may be prevented by elimination of calcium from the extracellular solution as well as by coexpression with the wild-type Kir 3.2 allele, or other members of the Kir 3.0 subfamily. These results suggest that the weaver defect in Kir 3.2 may cause cerebellar cell death by cell swelling and calcium overload. Cells which express the weaver subunit, but which normally survive, may do so because of heteromeric subunit assembly with wild-type subunits of the Kir 3.0 subfamily.


T Lymphocytes infiltrating various tumour types express the MHC class II ligand lymphocyte activation gene-3 (LAG-3): role of LAG-3/MHC class II interactions in cell-cell contacts.
The product of the Lymphocyte Activation Gene-3 (LAG-3, CD223) is a high affinity MHC class II ligand expressed by activated CD4(+) and CD8(+) T cells, which can associate with the T cell receptor (TCR) and downregulate TCR signalling in vitro. We have also reported that a soluble mLAG-3Ig fusion protein works as a vaccine adjuvant in vivo in mice, enhancing Th1 and CD8 T cell responses. Here, we report that LAG-3 expression was found, using fluorescent activated cell sorting (FACS) analysis, on 11-48% of human tumour-infiltrating lymphocytes (TILs) isolated from eight freshly dissociated renal cell carcinomas (RCCs), and was restricted mostly to CD8(+) cells. Immunohistochemical analysis confirmed LAG-3 expression by TILs in 9/11 RCCs, as well as in tumours of different origins, such as melanomas (3/5) and lymphomas (7/7). Since not only antigen presenting cells (APCs), but also TILs themselves strongly express major histocompatibility complex (MHC) class II, we firstly investigated whether LAG-3/MHC class II T-T cell contacts might influence tumour cell recognition. However, cytotoxicity inhibition was not observed in two RCC-specific CD8(+) T cell clones in the presence of the LAG-3-specific MAb, and there was also no observed difference in the recognition of LAG-3-transfected or wild-type RCC by these cytotoxic T lymphocytes (CTLs). In contrast, MHC class II engagement by LAG-3Ig was found to enhance the capacity of immature dendritic cells to stimulate naive T cell proliferation and IL-12-dependent IFN-gamma production by T cells in vitro. These results therefore provide support for a role for TIL-expressed LAG-3 in the engagement of class II molecules on APCs, thereby contributing to APC activation and Th1/Tc1 commitment, without downregulating cytotoxicity.


Coping with toothache pain: a qualitative study of low-income persons and minorities.
This study examined the behavioral impact of toothache pain as well as self-care strategies for pain relief among minority and low-income individuals. Eight focus group sessions were conducted with 66 participants drawn from low-income non-Hispanic White, non-Hispanic Black, and Hispanic adults over the age of 20 who had experienced a toothache during the previous 12-month period and who had utilized self-care or care from a nondentist. Toothache pain was described as intense, throbbing, miserable, or unbearable. Focus group participants indicated that toothache pain affected their ability to perform normal activities, such as their job, housework, social activities, sleeping, talking, and eating, as well as making them depressed and affecting their social interactions. Numerous prescription and nonprescription medications as well as home remedies and self-care strategies were used for pain relief, although these were generally of limited and uncertain benefit. While receiving care at a dental office was the most preferable option for care, most participants reported multiple barriers, including the cost of dental care that resulted in long delays in seeking dental care. The main reason for eventually seeking dental care was the severity of the pain. Although removing financial barriers alone may not lead to preventive dental visits, it would facilitate more timely visits to dentists to treat toothache pain.


Specificity of color connectivity between primate V1 and V2.
To examine the functional interactions between the color and form pathways in the primate visual cortex, we have examined the functional connectivity between pairs of color oriented and nonoriented V1 and V2 neurons in Macaque monkeys. Optical imaging maps for color selectivity, orientation preference, and ocular dominance were used to identify specific functional compartments within V1 and V2 (blobs and thin stripes). These sites then were targeted with multiple electrodes, single neurons isolated, and their receptive fields characterized for orientation selectivity and color selectivity. Functional interactions between pairs of V1 and V2 neurons were inferred by cross-correlation analysis of spike firing. Three types of color interactions were studied: nonoriented V1/nonoriented V2 cell pairs, nonoriented V1/oriented V2 cell pairs, and oriented V1/nonoriented V2 cell pairs. In general, interactions between V1 and V2 neurons are highly dependent on color matching. Different cell pairs exhibited differing dependencies on spatial overlap. Interactions between nonoriented color cells in V1 and V2 are dependent on color matching but not on receptive field overlap, suggesting a role for these interactions in coding of color surfaces. In contrast, interactions between nonoriented V1 and oriented V2 color cells exhibit a strong dependency on receptive field overlap, suggesting a separate pathway for processing of color contour information. Yet another pattern of connectivity was observed between oriented V1 and nonoriented V2 cells; these cells exhibited interactions only when receptive fields were far apart and failed to interact when spatially overlapped. Such interactions may underlie the induction of color and brightness percepts from border contrasts. Our findings thus suggest the presence of separate color pathways between V1 and V2, each with differing patterns of convergence and divergence and distinct roles in color and form vision.


The impact of using computer decision-support software in primary care nurse-led telephone triage: interactional dilemmas and conversational consequences.
Telephone triage represents one strategy to manage demand for face-to-face GP appointments in primary care. Although computer decision-support software (CDSS) is increasingly used by nurses to triage patients, little is understood about how interaction is organized in this setting. Specifically any interactional dilemmas this computer-mediated setting invokes; and how these may be consequential for communication with patients. Using conversation analytic methods we undertook a multi-modal analysis of 22 audio-recorded telephone triage nurse-caller interactions from one GP practice in England, including 10 video-recordings of nurses' use of CDSS during triage. We draw on Goffman's theoretical notion of participation frameworks to make sense of these interactions, presenting 'telling cases' of interactional dilemmas nurses faced in meeting patient's needs and accurately documenting the patient's condition within the CDSS. Our findings highlight troubles in the 'interactional workability' of telephone triage exposing difficulties faced in aligning the proximal and wider distal context that structures CDSS-mediated interactions. Patients present with diverse symptoms, understanding of triage consultations, and communication skills which nurses need to negotiate turn-by-turn with CDSS requirements. Nurses therefore need to have sophisticated communication, technological and clinical skills to ensure patients' presenting problems are accurately captured within the CDSS to determine safe triage outcomes. Dilemmas around how nurses manage and record information, and the issues of professional accountability that may ensue, raise questions about the impact of CDSS and its use in supporting nurses to deliver safe and effective patient care.


Women's health content validity of the Family Medicine In-training Examination.
Women's health is the field of medicine that addresses the special health care needs of women. It includes conditions more common, different, or unique to women. The primary means of knowledge assessment for family medicine residents is the Family Medicine In-training Examination (FMITE). This study describes the degree to which the FMITE tests residents for knowledge of women's health. This was a content validity study using item analysis to identify FMITE women's health questions. The curriculum standard was the 2004 American Academy of Family Physicians (AAFP) Women's Health Curriculum Guidelines. Two independent reviewers verified the method of identifying women's health questions. The test sample included 3,460 questions (1996-2005 FMITEs). Women's health questions made up 23% of the sample. Content areas and topics from the AAFP curriculum were sampled unevenly. The exam emphasized reproductive health and failed to assess knowledge of older women's health. The FMITE does test resident knowledge of the traditional subjects of obstetrics and gynecology but does not provide an adequate assessment of the breadth of knowledge of women's health.


A high oxidative stress index predicts endothelial dysfunction in young male smokers.
Experimental studies have shown that smoking was related to endothelial dysfunction via oxidative stress. However, the degree of oxidative stress to be associated with endothelial dysfunction is unknown. Oxidative stress index (OSI) might be a useful and easy way of determining the endothelial dysfunction. Hence, we aimed to evaluate the relationship between OSI and flow mediated dilatation (FMD) in smoking healthy male volunteers. Eighty smoking healthy male volunteers were enrolled in the study. Participants were classified as having normal and abnormal FMD response. In an univariate analysis; systolic and diastolic blood pressures, C-reactive protein (CRP), low-density lipoprotein cholesterol, OSI and lipid peroxidation (LPO) levels were predictive for abnormal FMD response. In a multivariable logistic regression analysis with forward stepwise method, OSI (OR: 3.194, 95% CI: 1.710-5.966, p<0.001) and CRP (OR: 2.082, 95% CI: 1.101-3.939, p 0.024) were found to be independent parameters for predicting abnormal FMD response in young male smokers. The optimal cut-off value of OSI for detecting abnormal FMD response was found to be >3.35, with 100 % sensitivity and 84.1 % specificity. We have shown that critical endothelial dysfunction can easily be detected by OSI in individuals, at risk for developing coronary artery disease, such as smokers (Tab. 3, Fig. 3, Ref. 30). Text in PDF www.elis.sk.


Breakthrough invasive candidiasis in patients on micafungin.
For Candida species, a bimodal wild-type MIC distribution for echinocandins exists, but resistance to echinocandins is rare. We characterized isolates from patients with invasive candidiasis (IC) breaking through >or=3 doses of micafungin therapy during the first 28 months of its use at our center: MICs were determined and hot-spot regions within FKS genes were sequenced. Eleven of 12 breakthrough IC cases identified were in transplant recipients. The median duration of micafungin exposure prior to breakthrough was 33 days (range, 5 to 165). Seventeen breakthrough isolates were recovered: FKS hot-spot mutations were found in 5 C. glabrata and 2 C. tropicalis isolates; of these, 5 (including all C. glabrata isolates) had micafungin MICs of >2 microg/ml, but all demonstrated caspofungin MICs of >2 microg/ml. Five C. parapsilosis isolates had wild-type FKS sequences and caspofungin MICs of 0.5 to 1 microg/ml, but 4/5 had micafungin MICs of >2 microg/ml. The remaining isolates retained echinocandin MICs of <or=2 microg/ml and wild-type FKS gene sequences. Breakthrough IC on micafungin treatment occurred predominantly in severely immunosuppressed patients with heavy prior micafungin exposure. The majority of cases were due to C. glabrata with an FKS mutation or wild-type C. parapsilosis with elevated micafungin MICs. MIC testing with caspofungin identified all mutant strains. Whether the naturally occurring polymorphism within the C. parapsilosis FKS1 gene responsible for the bimodal wild-type MIC distribution is also responsible for micafungin MICs of >2 microg/ml and clinical breakthrough or an alternative mechanism contributes to the nonsusceptible echinocandin MICs in C. parapsilosis requires further study.


Prevalence of macrolides-lincosamides-streptogramin B resistance and erm gene classes among clinical strains of staphylococci and streptococci.
A total of 332 staphylococcal and 263 streptococcal isolates from three hospital microbiology laboratories were tested with erythromycin, clindamycin, and vernamycin B alpha to determine the prevalence of macrolides-lincosamides-streptogramin B resistance. Constitutive resistance was detected in 28 Staphylococcus aureus isolates (15.5%), 53 coagulase-negative staphylococci (35.1%), and 20 streptococci (7.6%). Inducible resistance was observed in 13 S. aureus isolates (7.2%), 25 coagulase-negative staphylococci (16.6%), and 2 streptococci (0.8%). Eleven coagulase-negative staphylococci (7.3%) exhibited a novel phenotype, namely inducible resistance to erythromycin and vernamycin B alpha but not clindamycin. Among the staphylococci, two variants of the inducible phenotype detected with the agar diffusion assay correlated with the presence of classical ermA or ermC genes, respectively, by dot-blot analysis. The prevalence of the staphylococcal phenotypes were different in the hospitals surveyed, and there was an apparent inverse correlation between the resistance observed and the use of erythromycin in each hospital.


VHL Type 2B gene mutation moderates HIF dosage in vitro and in vivo.
Von Hippel-Lindau (VHL) disease is caused by germline mutations in the VHL tumor suppressor gene, with Type 2B missense VHL mutations predisposing to renal cell carcinoma, hemangioblastoma and pheochromocytoma. Type 2B mutant pVHL is predicted to be defective in hypoxia inducible factor (HIF)-alpha regulation. Murine embryonic stem (ES) cells in which the endogenous wild-type Vhl gene was replaced with the representative Type 2B VHL hotspot mutation R167Q (Vhl(2B/2B)) displayed preserved physiological regulation of both HIF factors with slightly greater normoxic dysregulation of HIF-2alpha. Differentiated Vhl(2B/2B)-derived teratomas overexpressed joint HIF targets Vegf and EglN3 but not the HIF-1alpha-specific target Pfk1. Vhl(2B/2B) teratomas additionally displayed a growth advantage over Vhl(-/-)-derived teratomas, suggestive of a tight connection between perturbations in the degree and ratio of HIF-1alpha and HIF-2alpha stabilization and cell growth. Vhl(2B/2B) mice displayed mid-gestational embryonic lethality, whereas adult Vhl(2B/+) mice exhibited susceptibility to carcinogen-promoted renal neoplasia compared with wild-type littermates at 12 months. Our experiments support a model in which the representative Type 2B R167Q mutant pVhl produces a unique profile of HIF dysregulation, thereby promoting tissue-specific effects on cell growth, development and tumor predisposition.


Cytoplasmic extraction: polyribosomes and heterogenous ribonucleoproteins without associated DNA.
Two methods are described for preparing cytoplasmic extracts from sea urchin embryos. One method, involving homogenization, yields DNA structures that cosediment with polyribosomes and subribosomal ribonucleoproteins. In addition this method also yields extraneous structures containing RNA that cosediment with polyribosomes. The DNA is not associated with polyribosomes, as shown by buoyant density analysis. Furthermore, this DNA appears to be spurious, because its release into a cytoplasmic extract does not occur when a different method of cell disruption, involving passage of embryos through a hypodermic needle, is used. With this second method, polyribosomes are obtained without extraneous cosedimenting RNA structures and subribosomal ribonucleo-proteins are obtained in the virtual absence of DNA.


Actual causes of death in Chaoyang District of Beijing, China, 2007.
To identify and quantify major external (non-genetic) factors that contribute to death in Chaoyang District of Beijing, China in 2007. The death registration data reported to the Center of Disease Control and Prevention of Chaoyang District of Beijing, China, during the year 2007, were obtained. The analysis was conducted in 2009 using the health risk factors identified by the World Health Report 2002 and the population attributable fractions of mortality from Global burden of disease and risk factors. The estimates of actual causes of death attributable to each risk factor were calculated by multiplying the population attributable fractions of mortality by the corresponding number of deaths of the subgroup or total population. The five leading actual causes of death in Chaoyang District of Beijing, China in 2007 were high blood pressure (2159 deaths, 18%), smoking (990, 8%), low fruit and vegetable consumption (968, 8%), high cholesterol (891, 7%), and physical inactivity (629, 5%). The pattern and ordering of these leading causes vary with sex and age specific subgroups. More than half of the total number of deaths in Chaoyang District in 2007 could be attributed to a few major preventable risk factors. Although the study focused on only one district of Beijing in one single year, and is by no means comprehensive, its findings suggest that public health policies and programmes in China should address these public health concerns by focusing on these largely preventable risk factors for primary prevention.


Programmed cell death (apoptosis) in lymphoid and myeloid cell lines during zinc deficiency.
Three human cell lines of lymphoid (Molt-3 and Raji) or myeloid (HL-60) origin were maintained in vitro under zinc-sufficient or zinc-deficient conditions. Under these conditions, cell proliferation, viability and mode of death (apoptotic or necrotic) were assessed. All three cell types decreased their proliferative capacity and viability under conditions of zinc deficiency. Cell death in the HL-60 and Raji cultures occurred primarily via apoptosis, while most cells in zinc-deficient Molt-3 cultures died via necrosis. Apoptosis in zinc-deficient cultures of HL-60 and Raji cells was characterized by a slow decline in culture viability as cells with condensed and fragmented nuclear DNA appeared. These morphological changes were accompanied by an increase in cell buoyant density, which allowed separation of viable apoptotic cells from their non-apoptotic counterparts by means of percoll stepdensity gradients. Necrosis in zinc-deficient Molt-3 cultures was characterized by rapid loss of cell culture viability as these cells underwent direct lysis. Intact necrotic cells were easily identified by the flocculated state of their chromatin as well as the decreased basophilia of their cytoplasm. Analysis of DNA from apoptotic HL-60 and Raji cells revealed that internucleosomal DNA degradation, indicative of endogenous endonuclease activation, had occurred, whereas the nuclear DNA of necrotic Molt-3 cells remained relatively unfragmented. The different modes of cell death evoked may reflect the relative sensitivities of cells of these lineages to zinc levels in vivo.


Medical audit in general practice. I: Effects on doctors' clinical behaviour for common childhood conditions. North of England Study of Standards and Performance in General Practice.
To estimate the effects of medical audit, particularly setting clinical standards, on general practitioners' clinical behaviour. Before and after study strengthened by a replicated Latin square. 62 training general practices in the north of England. 92 general practitioner trainers, 84 (91%) of whom completed the study; random sample of 3500 children consulting one of these trainers for any of five conditions--acute cough, acute vomiting, bedwetting, itchy rash, and recurrent wheezy chest--stratified by doctor consulted, condition, and age. Clinical standard set by each of 10 small groups of general practitioner trainers for one randomly selected childhood condition. Each group also experienced a different type of medical audit, randomly selected, for each of the four other study conditions (receiving a clinical standard set by another trainer group, tabulated data comparing clinical performance with that of all other groups, tabulated data from only their own group, and nothing ("control" condition)). Content of initial consultation divided into: history, examination, investigation, diagnosis, and management (abstracted from medical records and "enhancement forms" completed by doctors). There was increased prescribing of bronchodilators for acute cough, oral rehydration fluids for acute vomiting, antibiotics for itchy rash, and bronchodilators and oral steroids for recurrent wheezy chest and reduced prescribing of antibiotics for acute cough and recurrent wheezy chest and tricyclic antidepressants for bedwetting. Fewer children were "discharged." Each change was consistent with the standard and either limited to doctors who set a standard for that condition or significantly greater for them than all other doctors. Setting clinical standards improved prescribing and follow up.


Improving outcomes after pediatric cardiac arrest - the ICU-Resuscitation Project: study protocol for a randomized controlled trial.
Quality of cardiopulmonary resuscitation (CPR) is associated with survival, but recommended guidelines are often not met, and less than half the children with an in-hospital arrest will survive to discharge. A single-center before-and-after study demonstrated that outcomes may be improved with a novel training program in which all pediatric intensive care unit staff are encouraged to participate in frequent CPR refresher training and regular, structured resuscitation debriefings focused on patient-centric physiology. This ongoing trial will assess whether a program of structured debriefings and point-of-care bedside practice that emphasizes physiologic resuscitation targets improves the rate of survival to hospital discharge with favorable neurologic outcome in children receiving CPR in the intensive care unit. This study is designed as a hybrid stepped-wedge trial in which two of ten participating hospitals are randomly assigned to enroll in the intervention group and two are assigned to enroll in the control group for the duration of the trial. The remaining six hospitals enroll initially in the control group but will transition to enrolling in the intervention group at randomly assigned staggered times during the enrollment period. To our knowledge, this is the first implementation of a hybrid stepped-wedge design. It was chosen over a traditional stepped-wedge design because the resulting improvement in statistical power reduces the required enrollment by 9 months (14%). However, this design comes with additional challenges, including logistics of implementing an intervention prior to the start of enrollment. Nevertheless, if results from the single-center pilot are confirmed in this trial, it will have a profound effect on CPR training and quality improvement initiatives. ClinicalTrials.gov, NCT02837497 . Registered on July 19, 2016.


Schizophrenia through the carers' eyes: results of a European cross-sectional survey.
Schizophrenia is a serious mental disorder affecting approximately 29 million people worldwide. The ideal treatment and care of patients with schizophrenia should be provided by a multidisciplinary 'team' involving psychiatrists, nurses and other healthcare professionals, together with carers and patients. In light of the key role carers play in the care of patients with schizophrenia, the present survey was designed to assess the opinions of family members and friends of patients with schizophrenia across Europe and to ascertain their attitudes towards the illness, medication and adherence to medication. Among carers participating in this survey, there was widespread awareness of the issues involved in supporting patients with schizophrenia and the importance of their role in improving poor adherence to medication. Three differences in opinion emerged between the views of carers and psychiatrists; psychiatrists rely more on the patient themselves when assessing adherence than carers would recommend; in contrast to psychiatrists, many carers believe the illness itself contributes to non-adherence; two thirds of carers think that schizophrenia medication damages health (higher than estimated by psychiatrists). The findings from the present survey, taken together with the results from the Adherencia Terapéutica en la Esquizofrenia surveys of psychiatrists and nurses, support the need for a collaborative approach to the issue of treatment nonadherence. In particular, healthcare professionals should recognize the valuable contribution that family carers can make to improve treatment adherence and consequently clinical outcomes for patients with schizophrenia. Schizophrenia carries a significant burden for families providing care. The Adherencia Terapéutica en la Esquizofrenia (ADHES) carers' survey was designed to assess the opinions of family and friends of patients with schizophrenia across Europe and ascertain their attitudes towards the illness, medication and adherence to medication. A questionnaire-based cross-sectional survey of 138 carers across 16 European countries. Interpretation of results was based on a descriptive comparison of responses. Carers recognized the importance of medication to help patients get better (76%) and improve their quality of life (76%) and relationships (74%). Sixty-seven per cent believed medication damages general health. Sixty-five per cent reported that treatment adherence was a burden for patients. Thirty-eight per cent indicated that it was a daily struggle to get patients to take their medication. Fifty per cent perceived that medication administered every few weeks rather than daily was quite/very important. Ninety-three per cent agreed on the importance of family support to boost adherence, with education and information deemed important for families and patients. Carers rely less on the patient themselves when assessing adherence than psychiatrists. The burden faced by carers and patients in taking medication provides an opportunity for healthcare professionals to provide support in a multidisciplinary 'team' involving psychiatrists, nurses and carers.


Anti-protozoal activity of extracts from chicory (Cichorium intybus) against Cryptosporidium parvum in cell culture.
Cryptosporidium spp. are responsible for severe public health problems and livestock production losses. Treatment options are limited to only one drug available for human and bovine cryptosporidiosis, respectively, and both drugs exhibit only partial efficacy. Sesquiterpene lactones (SL) are plant bioactive compounds that function as a defence mechanism against herbivores. SL have demonstrated anti-parasitic properties against a range of parasitic taxa but knowledge about their anti-Cryptosporidium efficacy is limited. The effect of SL-rich leaf and root extracts from chicory (Cichorium intybus cv. Spadona) was investigated using human colon adenocarcinoma (HCT-8) cells infected with Cryptosporidium parvum. C. parvum oocysts were inoculated onto the cell monolayer and i) incubated for 4 hours with extracts (leaf and root extracts 300, 150, 75, 37.5, 18.75 and 9.375 μg/mL) in triplicates followed by incubation in bioactive free media (sporozoite invasion assays) or ii) incubated for 4 hours in bioactive free media followed by 48-hours incubation with extracts (growth inhibition assays). Extract toxicity on HCT-8 cells was assessed via water-soluble tetrazolium (WST)-1 assay prior to quantifying parasitic growth via immunofluorescence. Both extracts demonstrated dose-dependent inhibition in the growth inhibition assays (p = < 0.0001 for both extracts) but not in the invasion assays. Anti-parasitic activity did not appear to be solely related to SL content, with the extract with lower SL content (leaf) exhibiting higher inhibition at 300 μg/ml. However, given the limited treatment options available for Cryptosporidium spp., our study encourages further investigation into the use of chicory extracts to identify novel active compound(s) inhibiting these protozoa.


Synthesis of enantiopure chloroalcohols by enzymatic kinetic resolution.
3-Alkenyl and heteroaryl chloroalcohols have been obtained in excellent enantiomeric excess (>99%) by enzymatic kinetic resolution using the haloalcohol dehalogenase HheC. Yields were close to the theoretical maximum for all substrates employed. Furthermore, the applicability of this methodology on multigram scale has been established.


Pyometra presenting in conjunction with bowel cancer in a post-menopausal women: a case report.
This case describes a 71 year old, post-menopausal woman who developed vaginal discharge. This complaint ultimately led to the discovery of bowel cancer in conjunction with a large sterile pyometra.The pyometra was not due to genital malignancy. The most likely conclusion is that the pyometra may have arisen as an inflammatory response to the adjacent bowel pathology. This case report highlights the need for clinicians to consider non-gynaecological cancer as a possible cause for otherwise unexplained pyometra.


Adaptation to discomfort in personal protective devices: an example with safety helmets.
Discomfort in the use of personal protective devices (PPD) has been one of the chief causes of their non-use. A field trial using industrial helmets was carried out to ascertain whether by training and repeated wearing subjects could experience a significant adaptation to discomfort. Ten subjects took part in the trial in a tropical environment by wearing helmets repeatedly (6 h a day) for one month. Subjective evaluations of discomfort were made at the end of the 1st, 3rd, 5th, 14th and 30th day. It was revealed that complaints of discomfort, viz. hotness, heaviness, bad fit, etc., decreased markedly throughout the 30 day period. Though positive responses of adaptation to discomfort seem to reach an optimum towards 30 days, it is difficult to draw any conclusions on the optimum period of adaptation for each discomfort factor. In relation to inherent discomforts that are extremely difficult to overcome without compromising the protection efficiency of a PPD, the principle of adaptation seems to be a very important facet which has to be developed for an effective PPD programme.


Serendipitous Identification of a Covalent Activator of Liver Pyruvate Kinase.
Enzymes are effective biological catalysts that accelerate almost all metabolic reactions in living organisms. Synthetic modulators of enzymes are useful tools for the study of enzymatic reactions and can provide starting points for the design of new drugs. Here, we report on the discovery of a class of biologically active compounds that covalently modifies lysine residues in human liver pyruvate kinase (PKL), leading to allosteric activation of the enzyme (EC50 =0.29 μM). Surprisingly, the allosteric activation control point resides on the lysine residue K282 present in the catalytic site of PKL. These findings were confirmed by structural data, MS/MS experiments, and molecular modelling studies. Altogether, our study provides a molecular basis for the activation mechanism and establishes a framework for further development of human liver pyruvate kinase covalent activators.


Site specific insertion of a type I rDNA element into a unique sequence in the Drosophila melanogaster genome.
We describe a cloned segment of unique DNA from the Oregon R strain of Drosophila melanogaster that contains a short type I insertion of the kind principally found within rDNA. The predominant type I rDNA insertion is 5kb in length, but there are also a co-terminal sub-set of shorter type I elements that share a common right hand junction with the rDNA. The insertion that we now describe is another member of this sub-set. The right hand junction of the type I sequence with the unique DNA is identical to the right hand junction of the type I sequences with rDNA. There is no significant feature within the insertion sequence that could have determined the position of the left junction with the sequence into which it is inserted. Like the corresponding short type I insertions in rDNA, the insertion into the unique DNA is flanked on both sides by a duplicated sequence, which in this case is 10 base pairs long. The cloning of a sequence corresponding to the uninterrupted unique location was facilitated by the observation that the Karsnas strain of D. melanogaster contains only uninterrupted sequences of this kind. The duplicated sequence at the target site for the insertion is only present as a single copy in the uninterrupted DNA. The sequence of the target site for the insertion (ACTGTTCT) in the unique segment shows a striking homology to the target in rDNA (ACTGTCCC).


Serum matrix metalloproteinase 9 and colorectal neoplasia: a community-based evaluation of a potential diagnostic test.
A blood test may be a more acceptable routine colorectal cancer (CRC) screening test than faecal occult blood test, flexible sigmoidoscopy or colonoscopy, and could be safer and cheaper. We evaluated the accuracy of a serum matrix metalloproteinase (MMP9) test for CRC in a non-presenting symptomatic population. A cohort, aged 50-69 with lower gastrointestinal symptoms, was identified by community-based survey. Accuracy of serum MMP9 was assessed by comparison with colonoscopy. Logistic regression identified predictors of neoplasia and receiver operating characteristic curve analyses determined the cutoff to maximise the sensitivity. Data were available for 748 patients. Overall, 46 cases of neoplasia were identified. Univariate analysis demonstrated that demographic characteristics, behavioural factors, clinical symptoms and raised serum MMP9 concentration were all significantly associated with the presence of neoplasia. Our final logistic regression model had a sensitivity of 79% and specificity of 70%. We demonstrated a significant association between serum MMP9 concentration and the presence of neoplasia. Serum MMP9 levels are raised in those with cancer and high-risk adenomas, although MMP9 estimation is likely to have the greatest predictive utility when used as part of a panel of biomarkers. Further work is required to identify biomarkers that are sufficiently accurate for implementing into routine practice.


The incidence of MCI differs by subtype and is higher in men: the Mayo Clinic Study of Aging.
Although incidence rates for mild cognitive impairment (MCI) have been reported, few studies were specifically designed to measure the incidence of MCI and its subtypes using published criteria. We estimated the incidence of amnestic MCI (aMCI) and nonamnestic MCI (naMCI) in men and women separately. A population-based prospective cohort of Olmsted County, MN, residents ages 70-89 years on October 1, 2004, underwent baseline and 15-month interval evaluations that included the Clinical Dementia Rating scale, a neurologic evaluation, and neuropsychological testing. A panel of examiners blinded to previous diagnoses reviewed data at each serial evaluation to assess cognitive status according to published criteria. Among 1,450 subjects who were cognitively normal at baseline, 296 developed MCI. The age- and sex-standardized incidence rate of MCI was 63.6 (per 1,000 person-years) overall, and was higher in men (72.4) than women (57.3) and for aMCI (37.7) than naMCI (14.7). The incidence rate of aMCI was higher for men (43.9) than women (33.3), and for subjects with ≤12 years of education (42.6) than higher education (32.5). The risk of naMCI was also higher for men (20.0) than women (10.9) and for subjects with ≤12 years of education (20.3) than higher education (10.2). The incidence rates for MCI are substantial. Differences in incidence rates by clinical subtype and by sex suggest that risk factors for MCI should be investigated separately for aMCI and naMCI, and in men and women.


A critical analysis of Atoh7 (Math5) mRNA splicing in the developing mouse retina.
The Math5 (Atoh7) gene is transiently expressed during retinogenesis by progenitors exiting mitosis, and is essential for ganglion cell (RGC) development. Math5 contains a single exon, and its 1.7 kb mRNA encodes a 149-aa polypeptide. Mouse Math5 mutants have essentially no RGCs or optic nerves. Given the importance of this gene in retinal development, we thoroughly investigated the possibility of Math5 mRNA splicing by Northern blot, 3'RACE, RNase protection assays, and RT-PCR, using RNAs extracted from embryonic eyes and adult cerebellum, or transcribed in vitro from cDNA clones. Because Math5 mRNA contains an elevated G+C content, we used graded concentrations of betaine, an isostabilizing agent that disrupts secondary structure. Although approximately 10% of cerebellar Math5 RNAs are spliced, truncating the polypeptide, our results show few, if any, spliced Math5 transcripts exist in the developing retina (<1%). Rare deleted cDNAs do arise via RT-mediated RNA template switching in vitro, and are selectively amplified during PCR. These data differ starkly from a recent study (Kanadia and Cepko 2010), which concluded that the vast majority of Math5 and other bHLH transcripts are spliced to generate noncoding RNAs. Our findings clarify the architecture of the Math5 gene and its mechanism of action. These results have implications for all members of the bHLH gene family, for any gene that is alternatively spliced, and for the interpretation of all RT-PCR experiments.


First case of non-exposed endoscopic wall-inversion surgery with sentinel node basin dissection for early gastric cancer.
Non-exposed endoscopic wall-inversion surgery (NEWS) is a novel technique of endoscopic full-thickness resection without transluminal access mainly designed to treat gastric cancer. Here, we report a successful case of NEWS with sentinel node basin dissection (SNBD) for early gastric cancer (EGC) with the risk of lymph node metastasis. A 55-year-old female patient with a 2-cm, diffuse-type intramucosal EGC with ulceration was referred to our hospital for a less invasive gastrectomy based on sentinel node navigation surgery. After obtaining informed consent, NEWS with SNBD was applied. After placing mucosal markings, indocyanine green solution was injected endoscopically into the submucosa around the lesion to examine sentinel nodes (SNs). The SN basin (the area of the left gastric artery), including three stained SNs(#3), was dissected, and an intraoperative pathological diagnosis confirmed that no metastasis had occurred. Subsequently, NEWS was performed for the primary lesion. Serosal markings were placed laparoscopically, submucosal injection was added endoscopically, and circumferential sero-muscular incision and suturing were performed laparoscopically, with the lesion inverted toward the inside of the stomach. Finally, the circumferential mucosal incision was performed, and the lesion was retrieved perorally. The operation was finished in 270 min without complications. The patient was uneventfully discharged 10 days after the procedure. The final pathological diagnosis was coincident with the pre- and intraoperative assessment. We demonstrated the feasibility and safety of NEWS with SNBD with a favorable result. This surgical concept is expected to become a promising, minimally invasive, function-preserving surgery to cure cases of EGC that are possibly node-positive.


Assessing the effects of social environment on blood pressure and heart rates of baboons.
The objective of this publication is to report on the feasibility of using a tether system for obtaining data on blood pressure and heart rates of socially housed primates and to evaluate the extent to which housing environment alters cardiovascular responses (mean arterial blood pressure and heart rate). Blood pressure and heart rates of adult male baboons (Papio cynocephalus hamadryas) were evaluated over a 6 week period under three different housing conditions: social companion, individual, and socially unfamiliar. Social environment was manipulated in a specially designed cage that incorporated removable panels of either woven wire or solid sheet metal. The design of the cage permitted nonhuman primates to engage in species-typical social behaviors such as grooming and aggression. Using a tether and catheter system, we monitored cardiovascular physiology. We tested the hypothesis that individual housing, housing with social companions, and housing with social strangers would produce different mean arterial blood pressure and heart rate responses. Individual housing and housing with strangers produced resting mean arterial blood pressures that were elevated relative to blood pressure responses with social companions. Individual housing and housing with social strangers produced different patterns of cardiovascular response. Individual housing resulted in lowered heart rates and elevated blood pressures relative to the social companion condition. Housing with social strangers resulted in both elevated blood pressure and elevated heart rate, relative to the social companion condition. Responses observed during this study demonstrated the sensitivity of blood pressure and heart rates to differences in social environment.


The Prediction of Radiotherapy Toxicity Using Single Nucleotide Polymorphism-Based Models: A Step Toward Prevention.
Radiotherapy is a mainstay of cancer treatment, used in either a curative or palliative manner to treat approximately 50% of patients with cancer. Normal tissue toxicity limits the doses used in standard radiation therapy protocols and impedes improvements in radiotherapy efficacy. Damage to surrounding normal tissues can produce reactions ranging from bothersome symptoms that negatively affect quality of life to severe life-threatening complications. Improved ways of predicting, before treatment, the risk for development of normal tissue toxicity may allow for more personalized treatment and reduce the incidence and severity of late effects. There is increasing recognition that the cause of normal tissue toxicity is multifactorial and includes genetic factors in addition to radiation dose and volume of exposure, underlying comorbidities, age, concomitant chemotherapy or hormonal therapy, and use of other medications. An understanding of the specific genetic risk factors for normal tissue response to radiation has the potential to enhance our ability to predict adverse outcomes at the treatment-planning stage. Therefore, the field of radiogenomics has focused upon the identification of genetic variants associated with normal tissue toxicity resulting from radiotherapy. Innovative analytic methods are being applied to the discovery of risk variants and development of integrative predictive models that build on traditional normal tissue complication probability models by incorporating genetic information. Results from initial studies provide promising evidence that genetic-based risk models could play an important role in the implementation of precision medicine for radiation oncology through enhancing the ability to predict normal tissue reactions and thereby improve cancer treatment.


The relation between personal characteristics of physicians and practice location in Manitoba.
A questionnaire survey of 562 physicians in Manitoba who had graduated from the University of Manitoba was carried out to assess the effect of personal characteristics on choosing a practice location. The results closely resemble those of studies performed in the United States: the choice of a nonurban practice location is significantly more likely if the physicians and their spouses have nonurban backgrounds and if the physicians have had a nonurban preceptorship during undergraduate medical education. In this study practitioners who were male and whose fathers were farmers or health care professionals were also more likely to practise in nonurban areas. These findings will help in making physician distribution more equitable.


Human thymic epithelial cells express an endogenous lectin, galectin-1, which binds to core 2 O-glycans on thymocytes and T lymphoblastoid cells.
Thymic epithelial cells play a crucial role in the selection of developing thymocytes. Thymocyte-epithelial cell interactions involve a number of adhesion molecules, including members of the integrin and immunoglobulin superfamilies. We found that human thymic epithelial cells synthesize an endogenous lectin, galectin-1, which binds to oligosaccharide ligands on the surface of thymocytes and T lymphoblastoid cells. Binding of T lymphoblastoid cells to thymic epithelial cells was inhibited by antibody to galectin-1 on the epithelial cells, and by two antibodies, T305 and 2B11, that recognize carbohydrate epitopes on the T cell surface glycoproteins CD43 and CD45, respectively. T lymphoblastoid cells and thymocytes bound recombinant galectin-1, as demonstrated by flow cytometric analysis, and lectin binding was completely inhibited in the presence of lactose. The degree of galectin-1 binding to thymocytes correlated with the maturation stage of the cells, as immature thymocytes bound more galectin-1 than did mature thymocytes. Preferential binding of galectin-1 to immature thymocytes may result from regulated expression of preferred oligosaccharide ligands on those cells, since we found that the epitope recognized by the T305 antibody, the core 2 O-glycan structure on CD43, was expressed on cortical, but not medullary cells. The level of expression of the UDP-GlcNAc:Gal beta 1,3GalNAc-R beta 1, 6GlcNAc transferase (core 2 beta 1, 6 GlcNAc transferase, or C2GnT), which creates the core 2 O-glycan structure, correlated with the glycosylation change between cortical and medullary cells. Expression of mRNA encoding the C2GnT was high in subcapsular and cortical thymocytes and low in medullary thymocytes, as demonstrated by in situ hybridization. These results suggest that galectin-1 participates in thymocyte-thymic epithelial cell interactions, and that this interaction may be regulated by expression of relevant oligosaccharide ligands on the thymocyte cell surface.


Intraarterial papaverine for the treatment of vasospasm.
The authors describe the use of intraarterial papaverine to treat vasospasm following subarachnoid hemorrhage. Two cases are reported: a 40-year-old woman with a posterior communicating artery aneurysm and a 67-year-old man with a posterior cerebral artery aneurysm. Both patients developed symptomatic, angiographically demonstrated vasospasm that responded to papaverine infusion.


Trust in micro-health insurance: an exploratory study in Rwanda.
Micro-health insurance (MHI) has been proposed as an alternative to user fees to improve access to medical care without the threat of financial impoverishment. To enrol in health insurance and pay a premium, consumers should be able to trust that insurers use their funds to reimburse providers who will deliver quality care when needed. In 1998, shortly after the re-introduction of user fees, the Rwandan Ministry of Health in close collaboration with the local population developed and implemented MHI in three districts. Member benefits cover preventive and curative care in health centres and ambulance transport to the district hospital where a limited package of services is also covered by MHI. This paper examines trust-building structures and practices in MHI in Rwanda. It is an exploratory, descriptive analysis using information collected from focus groups. Findings suggest that MHI managers, providers and policy-makers need to think systematically about a wide range of initiatives that enhance trust and caring, and to design trust-building structures and practices in the consumer-insurance-provider arrangement.


Dissecting structural transitions in the HIV-1 dimerization initiation site RNA using 2-aminopurine fluorescence.
A highly conserved 35 nucleotide RNA stem-loop, the dimerization initiation site (DIS), in the 5' untranslated region (UTR) of the human immunodeficiency virus type I (HIV-1) genome has been identified as the sequence primarily responsible for initiation of viral genome dimerization. The DIS initiates viral genome dimerization through a loop-loop 'kissing' interaction and is converted from an intermediate 'kissing' to a more thermodynamically stable extended duplex dimer in a conformational rearrangement that is chaperoned by the HIV-1 nucleocapsid protein (NCp7). Here we describe fluorescence methods designed to probe local RNA dynamics and structural transitions associated with the DIS dimer formation and its NCp7 chaperoned structural conversion. These methods take advantage of the exquisite sensitivity of the quantum yield of the fluorescent nucleotide base analog, 2-aminopurine (2-AP), to its immediate structural and dynamic environment. The 2-AP fluorescence methods described allow a detailed kinetic and thermodynamic examination of this type of RNA-RNA interaction, as well as an analysis of the molecular mechanism of NCp7 chaperone activity.


Mammalian S-phase checkpoint integrity is dependent on transformation status and purine deoxyribonucleosides.
In eukaryotic cells arrested in S-phase, checkpoint controls normally restrain mitosis until after replication. We have identified an array of previously unsuspected factors that modulate this restraint, using transformed hamster cells in which cycle controls are known to be altered in S-phase arrest. Arrested cells accumulate cyclin B, the regulatory partner of the mitotic p34(cdc2) kinase, which is normally not abundant until late G(2) phase; treatment of arrested cells with caffeine produces rapid S-phase condensation. We show here that such S-phase checkpoint slippage, as visualised through caffeine-dependent S-phase condensation, correlates with rodent origin and transformed status, is opposed by reverse transformation, and is favoured by c-src and opposed by wnt1 overexpression. Slippage is also dependent on a prolonged replicative arrest, and is favoured by arrest with hydroxyurea, which inhibits ribonucleotide reductase. This last is a key enzyme in deoxyribonucleotide synthesis, recently identified as a determinant of malignancy. Addition of deoxyribonucleosides shows that rapid S-phase condensation is suppressed by a novel checkpoint mechanism: purine (but not pyrimidine) deoxyribonucleosides, like reverse transformation, suppress cyclin B/p34(cdc2) activation by caffeine, but not cyclin B accumulation. Thus, ribonucleotide reductase has an unexpectedly complex role in mammalian cell cycle regulation: not only is it regulated in response to cycle progression, but its products can also reciprocally influence cell cycle control kinase activation.


A highly conserved domain of the maize activator transposase is involved in dimerization.
Previous studies have presented indirect evidence that the transposase of the maize transposable element Activator (TPase) is active as an oligomer and forms inactive macromolecular complexes expressed in large amounts. Here, we have identified and characterized a dimerization domain at the C terminus of the protein. This domain is the most highly conserved region in the transposases of elements belonging to the Activator superfamily (hAT element superfamily) and contains a characteristic signature motif. The isolated dimerization domain forms extremely stable dimers in vitro. Interestingly, mutations in five of the six conserved residues of the signature motif do not affect in vitro dimerization, whereas mutations in other, less strictly conserved residues of the signature motif do. Loss of dimerization in vitro correlates with loss of TPase activity in vivo. As revealed by in situ immunofluorescence staining of mutant TPase proteins, the dimerization domain also is involved in forming inactive macromolecular aggregates when overexpressed, and the TPase contains one or more additional interaction functions.


Characterisation of the murine Hox-3.3 gene and its promoter.
The murine Hox-3.3 homeobox containing gene and its Xenopus homologue (XlHbox 1) produce two embryonic transcripts from two distinct promoters located approximately 9 kb apart. In order to begin to characterise one of these promoter regions (PRII), we have sequenced 3 kb of DNA immediately upstream of the transcription start site of the PRII transcript and analyzed the sequence for sequences known to bind transcription factors. Within this region are located a number of sequences that match known cis-elements. We have analysed the ability of two of these sequences that match to the Drosophila hunchback and Antennapedia/fushi-tarazu consensus binding sequences to specifically bind proteins extracted from embryos and from adult tissues. Using gel retention assays with oligonucleotides derived from these sequences, we show that both sequences specifically bind proteins present in extracts of mouse embryos and some, but not all extracts of various adult tissues. Protein binding cannot, however, be correlated with the known spatial domains of Hox-3.3 expression, suggesting that binding to these sequences is not simply related to activation of Hox-3.3 expression. A two base pair change in the most conserved region of the hunchback-like binding sequence completely abolishes protein binding. The presence of these highly conserved cis-acting elements that are known to be involved in regulation of the hunchback, even-skipped and engrailed genes in Drosophila suggests that these sequences may also be involved in the regulation of expression of Hox-3.3 and furthermore that regulation may in part at least involve binding of hunchback-like proteins (i.e. zinc-finger proteins) and Antennapedia-like homeobox-containing proteins.


Diffusion tensor smoothing through weighted Karcher means.
Diffusion tensor magnetic resonance imaging (MRI) quantifies the spatial distribution of water Diffusion at each voxel on a regular grid of locations in a biological specimen by Diffusion tensors- 3 × 3 positive definite matrices. Removal of noise from DTI is an important problem due to the high scientific relevance of DTI and relatively low signal to noise ratio it provides. Leading approaches to this problem amount to estimation of weighted Karcher means of Diffusion tensors within spatial neighborhoods, under various metrics imposed on the space of tensors. However, it is unclear how the behavior of these estimators varies with the magnitude of DTI sensor noise (the noise resulting from the thermal e!ects of MRI scanning) as well as the geometric structure of the underlying Diffusion tensor neighborhoods. In this paper, we combine theoretical analysis, empirical analysis of simulated DTI data, and empirical analysis of real DTI scans to compare the noise removal performance of three kernel-based DTI smoothers that are based on Euclidean, log-Euclidean, and affine-invariant metrics. The results suggest, contrary to conventional wisdom, that imposing a simplistic Euclidean metric may in fact provide comparable or superior noise removal, especially in relatively unstructured regions and/or in the presence of moderate to high levels of sensor noise. On the contrary, log-Euclidean and affine-invariant metrics may lead to better noise removal in highly structured anatomical regions, especially when the sensor noise is of low magnitude. These findings emphasize the importance of considering the interplay of sensor noise magnitude and tensor field geometric structure when assessing Diffusion tensor smoothing options. They also point to the necessity for continued development of smoothing methods that perform well across a large range of scenarios.


Perceived speed differences explain apparent compression in slit viewing.
When a figure moves behind a stationary narrow slit, observers often report seeing the figure as an integrated whole, a phenomenon known as slit viewing or anorthoscopic perception. Interestingly, in slit viewing, the figure is perceived compressed along the axis of motion, e.g., a circle is perceived as an ellipse. Underestimation of the speed of the moving object was offered as an explanation for this apparent compression. We measured perceived speed and compression in anorthoscopic perception and found results that are inconsistent with this hypothesis. We found evidence for an alternative hypothesis according to which apparent compression results from perceived speed differences between different parts of the figure, viz., the trailing parts are perceived to move faster than the leading parts. These differences in the perceived speeds of the trailing and the leading edges may be due to differences in the visibilities of the leading and trailing parts. We discuss our findings within a non-retinotopic framework of form analysis for moving objects.


MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes.
Antidepressants (ADs) are the most common treatment for major depressive disorder (MDD). However, only ∼30% of patients experience adequate response after a single AD trial, and this variability remains poorly understood. Here, we investigated microRNAs (miRNAs) as biomarkers of AD response using small RNA-sequencing in paired samples from MDD patients enrolled in a large, randomized placebo-controlled trial of duloxetine collected before and 8 weeks after treatment. Our results revealed differential expression of miR-146a-5p, miR-146b-5p, miR-425-3p and miR-24-3p according to treatment response. These results were replicated in two independent clinical trials of MDD, a well-characterized animal model of depression, and post-mortem human brains. Furthermore, using a combination of bioinformatics, mRNA studies and functional in vitro experiments, we showed significant dysregulation of genes involved in MAPK/Wnt signalling pathways. Together, our results indicate that these miRNAs are consistent markers of treatment response and regulators of the MAPK/Wnt systems.


Quantitative breast mass classification based on the integration of B-mode features and strain features in elastography.
Elastography is a new sonographic imaging technique to acquire the strain information of tissues and transform the information into images. Radiologists have to observe the gray-scale distribution of tissues on the elastographic image interpreted as the reciprocal of Young's modulus to evaluate the pathological changes such as scirrhous carcinoma. In this study, a computer-aided diagnosis (CAD) system was developed to extract quantitative strain features from elastographic images to reduce operator-dependence and provide an automatic procedure for breast mass classification. The collected image database was composed of 45 malignant and 45 benign breast masses. For each case, tumor segmentation was performed on the B-mode image to obtain tumor contour which was then mapped to the elastographic images to define the corresponding tumor area. The gray-scale pixels around tumor area were classified into white, gray, and black by fuzzy c-means clustering to highlight stiff tissues with darker values. Quantitative strain features were then extracted from the black cluster and compared with the B-mode features in the classification of breast masses. The performance of the proposed strain features achieved an accuracy of 80% (72/90), a sensitivity of 80% (36/45), a specificity of 80% (36/45), and a normalized area under the receiver operating characteristic curve, Az=0.84. Combining the strain features with the B-mode features obtained a significantly better Az=0.93, p-value<0.05. Summarily, the quantified strain features can be combined with the B-mode features to provide a promising suggestion in distinguishing malignant from benign tumors.