Novel LC-ESI/MS/MS(n) method for the characterization and quantification of 2'-deoxyguanosine adducts of the dietary carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine by 2-D linear quadrupole ion trap mass spectrometry.
An accurate and sensitive liquid chromatography-electrospray ionization/multi-stage mass spectrometry (LC-ESI/MS/MS(n)) technique has been developed for the characterization and quantification of 2'-deoxyguanosine (dG) adducts of the dietary mutagen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). PhIP is an animal and potential human carcinogen that occurs in grilled meats. Following enzymatic digestion and adduct enrichment by solid-phase extraction (SPE), PhIP-DNA adducts were analyzed by MS/MS and MS(n) scan modes on a 2-D linear quadrupole ion trap mass spectrometer (QIT/MS). The major DNA adduct, N-(deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (dG-C8-PhIP), was detected in calf thymus (CT) DNA modified in vitro with a bioactivated form of PhIP and in the colon and liver of rats given PhIP as part of the diet. The lower limit of detection (LOD) was 1 adduct per 10(8) DNA bases, and the limit of quantification (LOQ) was 3 adducts per 10(8) DNA bases in both MS/MS and MS(3) scan modes, using 27 microg of DNA for analysis. Measurements were based on isotope dilution with the internal standard, N-(deoxyguanosin-8-yl)-2-amino-1-(trideutero)methyl-6-phenylimidazo[4,5-b]pyridine (dG-C8-[2H3C]-PhIP). The selected reaction monitoring (SRM) scan mode in MS/MS was employed to monitor the loss of deoxyribose (dR) from the protonated molecules of the adducts ([M + H - 116]+). The consecutive reaction monitoring (CRM) scan modes in MS(3) and MS(4) were used to measure and further characterize product ions of the aglycone ion (BH2+) (Guanyl-PhIP). The MS(3) scan mode was effective in eliminating isobaric interferences observed in the MS/MS scan mode and resulted in an improved signal-to-noise (S/N) ratio. Moreover, the product ion spectra obtained by the MS(n) scan modes provided rich structural information about the adduct and were used to corroborate the identity of dG-C8-PhIP. In addition, an isomeric dG-PhIP adduct was detected in vivo. This LC-ESI/MS/MS(n) method is the first reported application on the use of the MS(3) scan mode for the analysis of DNA adducts in vivo.


The hrpZ gene of Pseudomonas syringae pv. phaseolicola enhances resistance to rhizomania disease in transgenic Nicotiana benthamiana and sugar beet.
To explore possible sources of transgenic resistance to the rhizomania-causing Beet necrotic yellow vein virus (BNYVV), Nicotiana benthamiana plants were constructed to express the harpin of Pseudomonas syringae pv. phaseolicola (HrpZ(Psph)). The HrpZ protein was expressed as an N-terminal fusion to the PR1 signal peptide (SP/HrpZ) to direct harpin accumulation to the plant apoplast. Transgene integration was verified by mPCR in all primary transformants (T0), while immunoblot analysis confirmed that the protein HrpZ(Psph) was produced and the signal peptide was properly processed. Neither T0 plants nor selfed progeny (T1) showed macroscopically visible necrosis or any other macroscopic phenotypes. However, plants expressing the SP/HrpZ(Psph) showed increased vigor and grew faster in comparison with non-transgenic control plants. Transgenic resistance was assessed after challenge inoculation with BNYVV on T1 progeny by scoring of disease symptoms and by DAS-ELISA at 20 and 30 dpi. Transgenic and control lines showed significant differences in terms of the number of plants that became infected, the timing of infection and the disease symptoms displayed. Plants expressing the SP/HrpZ(Psph) developed localized leaf necrosis in the infection area and had enhanced resistance upon challenge with BNYVV. In order to evaluate the SP/HrpZ-based resistance in the sugar beet host, A. rhizogenes-mediated root transformation was exploited as a transgene expression platform. Upon BNYVV inoculation, transgenic sugar beet hairy roots showed high level of BNYVV resistance. In contrast, the aerial non-transgenic parts of the same seedlings had virus titers that were comparable to those of the seedlings that were untransformed or transformed with wild type R1000 cells. These findings indicate that the transgenically expressed SP/HrpZ protein results in enhanced rhizomania resistance both in a model plant and sugar beet, the natural host of BNYVV. Possible molecular mechanisms underlying the enhanced resistance and plant growth phenotypes observed in SP/HrpZ transgenic plants are discussed.


Comparison of estrogen receptors in human premenopausal and postmenopausal uteri: indication of biologically inactive receptor in postmenopausal uteri.
Analysis of the estrogen receptor (ER) from uteri of premenopausal and postmenopausal patients has largely been limited to quantitation. The present study examined the biologic activity of the receptors with regard to their nuclear binding capacities. A Scatchard analysis of myometrial cytosol indicated high-affinity and low-capacity estradiol binding (Kd = 10(-10 M). A 35% ammonium sulfate fraction of cytosol was characterized for steroid specificity, receptor concentration, isoelectric focusing patterns, and capacity of the ER to bind to nuclear acceptor sites in a cell-free assay. The ER from premenopausal uteri displayed an extensive but saturable binding to the nuclear acceptor sites. In contrast, the postmenopausal ER displayed minimal binding. These cell-free binding differences were supported by tissue culture studies. This difference may be accentuated by the fact that much of the receptor is bound by endogenous estrogen in the premenopausal uterine preparation, which would decrease the subsequent binding of the [3H]ER to nuclear sites. This receptor in the postmenopausal uterus is largely unbound and thus would not compete for nuclear sites. These studies suggest that a significant fraction of the ER in postmenopausal uteri is biologically inactive; for example, it cannot translocate and bind to nuclear acceptor sites. This loss in nuclear binding may represent a change in the ER occurring at menopause as a result of changing endocrine states.


3D microtumors in vitro supported by perfused vascular networks.
There is a growing interest in developing microphysiological systems that can be used to model both normal and pathological human organs in vitro. This "organs-on-chips" approach aims to capture key structural and physiological characteristics of the target tissue. Here we describe in vitro vascularized microtumors (VMTs). This "tumor-on-a-chip" platform incorporates human tumor and stromal cells that grow in a 3D extracellular matrix and that depend for survival on nutrient delivery through living, perfused microvessels. Both colorectal and breast cancer cells grow vigorously in the platform and respond to standard-of-care therapies, showing reduced growth and/or regression. Vascular-targeting agents with different mechanisms of action can also be distinguished, and we find that drugs targeting only VEGFRs (Apatinib and Vandetanib) are not effective, whereas drugs that target VEGFRs, PDGFR and Tie2 (Linifanib and Cabozantinib) do regress the vasculature. Tumors in the VMT show strong metabolic heterogeneity when imaged using NADH Fluorescent Lifetime Imaging Microscopy and, compared to their surrounding stroma, many show a higher free/bound NADH ratio consistent with their known preference for aerobic glycolysis. The VMT platform provides a unique model for studying vascularized solid tumors in vitro.


Coronavirus-Associated Coagulopathy: Lessons From SARS-CoV1 and MERS-CoV for the Current SARS-CoV2 Pandemic.
To date, several studies have suggested a severe acute respiratory syndrome coronavirus 2 (SARS-CoV2)-mediated hypercoagulability in the forms of pulmonary embolism, stroke, gangrene, "COVID toes," as well as other acute thrombotic complications, warranting the use of systemic anticoagulation. Currently, there are no definitive recommendations as to the timing and dosing of prophylactic or therapeutic anticoagulation in coronavirus disease 2019 (COVID-19) patients. In this manuscript, we report a case of SARS-CoV2-mediated hypercoagulability and review the literature pertaining to the incidence and pathophysiology of coronavirus-mediated coagulopathies. A 64-year-old female, with a medical history of hypothyroidism and remote tobacco abuse, presented to the ED with fever and nonproductive cough. She had multiple negative SARS-CoV2 nasopharyngeal PCR tests during her hospital stay, but chest imaging and elevated inflammatory markers were suggestive of SARS-CoV2 infection. Computed tomography showed a left upper lobe pulmonary embolism with associated right heart strain, and an enlargement of the main pulmonary artery, for which she was initiated on therapeutic anticoagulation with low molecular weight heparin. Despite the medical management of her pulmonary embolism and conservative management of her SARS-CoV2, her clinical condition worsened requiring intubation and mechanical ventilation. After seven days, she was successfully extubated and was transferred to the medical service where her clinical course remained stable and subsequently discharged home on apixaban. In patients with SARS-CoV1-, SARS-CoV2-, and the Middle East respiratory syndrome coronavirus (MERS-CoV)-mediated hypercoagulability, the risk of thrombosis appears to be multifactorial - direct viral cytopathological effects, a pro-inflammatory state, cytokine storm, hypoxia-inducible thrombosis, and endothelial inflammation culminating in the formation of intra-alveolar or systemic fibrin clots. While initial guidelines have been developed to assist clinicians in selecting appropriate chemoprophylaxis as well as therapeutic anticoagulation, a consensus statement remains lacking. Further studies are needed to evaluate the pathogenesis and treatment of coronavirus-induced thrombosis.


Understanding mechanisms of change in a family-based preventive mental health intervention for refugees by refugees in New England.
Transnational migration of refugees is associated with poor mental health, particularly among children. We conducted a pilot trial of the Family Strengthening Intervention for Refugees (FSI-R), using a community-based participatory research (CBPR) approach to deliver a home-based intervention "for refugees by refugees" to improve family functioning and child mental health. N = 80 refugee families in the Greater Boston area participated in the study (n = 40 Somali Bantu families; n = 40 Bhutanese families) with n = 41 families randomized to care-as-usual. Of the 39 families who received FSI-R, n = 36 caregivers and children completed qualitative exit interviews. We present findings from these interviews to identify the mechanisms through which a family-strengthening intervention for refugees can be acceptable, feasible, and effective at improving family functioning and children's mental health outcomes. Authors applied Grounded Theory to code interview transcripts and detailed field notes and used an iterative process to arrive at final codes, themes, and a theoretical framework. The greatest contributors to acceptability and feasibility included flexibility in scheduling intervention sessions, the interventionist being a community member, and improvements to family communication and time spent together. All of these factors were made possible by the CBPR approach. Our findings suggest that given the socio-political context within the U.S. and the economic challenges faced by refugee families, the successful implementation of such interventions hinges on culturally-grounding the intervention design process, drawing heavily on community input, and prioritizing community members as interventionists.


A Significant Increasing Risk Association between Cigarette Smoking and XPA and XPC Genes Polymorphisms.
Cigarette smoking (CS) is a major cause of various serious diseases due to tobacco chemicals. There is evidence suggesting that CS has been linked with the DNA damage repair system, as it can affect genomic stability, inducing genetic changes in the genes involved in the repair system, specifically the nucleotide excision repair (NER) pathway, affecting the function and/or regulation of these genes. Single nucleotide polymorphism (SNP), along with CS, can affect the work of the NER pathway and, therefore, could lead to different diseases. This study explored the association of four SNPs in both XPA and XPC genes with CS in the Saudi population. The Taq Man genotyping assay was used for 220 healthy non-smokers (control) and 201 healthy smokers to evaluate four SNPs in the XPA gene named rs10817938, rs1800975, rs3176751, and rs3176752 and four SNPs in the XPC gene called rs1870134, rs2228000, rs2228001, and rs2607775. In the XPA gene, SNP rs3176751 showed a high-risk association with CS-induced diseases with all clinical parameters, including CS duration, CS intensity, gender, and age of smokers. On the other hand, SNP rs1800975 showed a statistically significant low-risk association with all clinical parameters. In addition, rs10817938 showed a high-risk association only with long-term smokers and a low-risk association only with younger smokers. A low-risk association was found in SNP rs3176752 with older smokers. In the XPC gene, SNP rs2228001 showed a low-risk association only with female smokers. SNP rs2607775 revealed a statistically significant low-risk association with CS-induced diseases, concerning all parameters, except for male smokers. However, SNP rs2228000 and rs1870134 showed no association with CS. Overall, the study results demonstrated possible significant associations (effector/and protector) between CS and SNPs polymorphisms in DNA repair genes, such as XPA and XPC, except for rs2228000 and rs1870134 polymorphisms.


[Risk factors of mental disorders in epileptic patients: case-control study].
The aim of this study was to analyse the risk factors of mental disorders in epileptic patients. It was a case-control study concerning 200 epileptic out-patients examined in the department of Neurology of Monastir University Hospital during four months. Patients had been divided into two groups: the first (study-group) included 100 epileptic patients with mental disorders compared with a control-group of 100 epileptic patients without mental disorders. General seizures were significantly more frequent in the study-group (78%) than in the control-group (57%) (p=0,001). In the study-group, mental disorders were dominated by the behavioraland character disorders. Existence of temporal focus in electroencephalography or structural abnormality in tomodensitometry was more frequent in epileptic patients with mental disorders. The medium term course of epilepsy was significantly longer in the study-group (15.38 years versus 10?35 years; p=0.00007). Association of antiepileptic agents was significantly more frequent in study-group (38% versus 15%; p= 0,0002). However, no correlations were found between mental disorders and sex, age at onset of epileptic seizures and seizures frequency. The results of our study suggest that epilepsy characteristics and anti-epileptic treatment seem to be the principal factors of mental disorders in epileptic patients.


The biosynthetic pathway of vitamin C in higher plants.
Vitamin C (L-ascorbic acid) has important antioxidant and metabolic functions in both plants and animals, but humans, and a few other animal species, have lost the capacity to synthesize it. Plant-derived ascorbate is thus the major source of vitamin C in the human diet. Although the biosynthetic pathway of L-ascorbic acid in animals is well understood, the plant pathway has remained unknown-one of the few primary plant metabolic pathways for which this is the case. L-ascorbate is abundant in plants (found at concentrations of 1-5 mM in leaves and 25 mM in chloroplasts) and may have roles in photosynthesis and transmembrane electron transport. We found that D-mannose and L-galactose are efficient precursors for ascorbate synthesis and are interconverted by GDP-D-mannose-3,5-epimerase. We have identified an enzyme in pea and Arabidopsis thaliana, L-galactose dehydrogenase, that catalyses oxidation of L-galactose to L-galactono-1,4-lactone. We propose an ascorbate biosynthesis pathway involving GDP-D-mannose, GDP-L-galactose, L-galactose and L-galactono-1,4-lactone, and have synthesized ascorbate from GDP-D-mannose by way of these intermediates in vitro. The definition of this biosynthetic pathway should allow engineering of plants for increased ascorbate production, thus increasing their nutritional value and stress tolerance.


In vitro safety and performance evaluation of a seawater solution enriched with copper, hyaluronic acid, and eucalyptus for nasal lavage.
The common cold is a viral infectious disease with symptoms such as runny nose, sore throat, and mainly, nasal congestion. State-of-the-art therapeutic approaches focus on alleviating the symptoms of this disease by non-invasive and simple-to-use methods. Nasal irrigation is one of the most accepted approaches to ease nasal congestion which, if left untreated, has a negative impact on the quality of life of patients. In this study, the safety and efficacy of a novel hypertonic seawater solution for nasal lavage enriched with hyaluronic acids, eucalyptus oil, copper, and manganese salts (Stérimar Stop & Protect Cold and Flu; SSPCF) have been investigated in vitro. An in vitro 3D reconstituted human nasal epithelium tissue model, MucilAir™, has been used in this study to investigate the safety of SSPCF on nasal epithelium by measuring transepithelial electrical resistance (TEER), lactate dehydrogenase (LDH), and interleukin-8 (IL-8) secretion. The efficacy of SSPCF was measured by mucociliary clearance (MCC), ATP release, Alcian blue and aquaporin (AQP3) stainings. SSPCF treatment respected nasal epithelium tissue integrity and enhanced barrier function without inducing a cytotoxic response. Secreted LDH and IL-8 levels were similar to untreated controls. MCC rate was increased 2.5-fold and ATP release decreased 87% upon SSPCF treatment, indicating improved decongestion activity. SSPCF treatment after hypotonic stress helped recover cellular organization, as shown by Alcian blue and AQP3 staining assays. SSPCF appears as a safe and effective nasal irrigation formula that may alleviate the symptoms associated with common cold such as nasal congestion.


Genotypic and phenotypic characterization of the Drosophila melanogaster olfactory mutation Indifferent.
Two Drosophila melanogaster third chromosomes carrying the EMS-induced mutations IndifferentA (IndfA) and IndifferentB (IndfB), previously isolated from larvae showing an anosmia when stimulated with nonanol, were recombined with a multi-marked chromosome in order to localize the mutant character(s). Recombinant strains were tested for their larval olfactory responses and classed as either mutant or wild type; both Indf characters were found to be located on the right arm of the chromosome, between ebony and claret. Deletion mapping suggests that the Indifferent wild-type character is a haplo-insufficiency and that IndfA and IndfB are located in cytological region 96A2-7. Deficiencies and both mutant strains were tested with 14 closely related odors (alcohols, acetates, acids and methyl esters, between eight and 10 carbons long). When stimulated with methyl octanoate, IndfA and IndfB appeared recessive; noncomplementation was observed for this phenotype in IndfA/IndfB hybrids indicating that the two characters are allelic. The overall responses of IndfA, IndfB and the deficiencies indicate that Indf is involved in processing organic odors of between eight and 10 carbons in length.


Infectious disease hospitalizations among infants in the United States.
This study describes the burden and epidemiologic features of infectious disease hospitalizations among infants in the United States. Hospitalizations with an infectious disease listed as a primary diagnosis for infants (<1 year of age) in the United States during 2003 were examined by using the Kids' Inpatient Database. National estimates of infectious disease hospitalizations, hospitalization rates, and various hospital parameters were examined. During 2003, an estimated 286,739 infectious disease hospitalizations occurred among infants in the United States and accounted for 42.8% of all infant hospitalizations. The national infectious disease hospitalization rate was 7010.8 hospitalizations per 100,000 live births, or approximately 1 infectious disease hospitalization for every 14 infants. The median length of stay was 3 days, and stays totaled >1 million hospital days for infants. Infectious disease hospitalization rates were highest among boys and nonwhite infants. The most commonly listed diagnoses among the infant infectious disease hospitalizations included lower respiratory tract infections (59.0%), kidney, urinary tract, and bladder infections (7.6%), upper respiratory tract infections (6.5%), and septicemia (6.5%). The median cost of an infectious disease hospitalization was $2235, with total annual hospital costs of approximately $690 million, among infants in the United States. Infectious disease hospitalizations among infants account for substantial health care expenditures and hospital time in the United States, with respiratory disease hospitalizations constituting more than one half of all hospitalizations. Younger infants, boys, and nonwhite infants were at increased risk for infectious disease hospitalization. Measures to reduce racial disparities and the occurrence of respiratory tract infections should substantially decrease the infectious disease burden among infants.


Longitudinal observation of influence of "taspo" on smoking behavior among high school students.
A system with an adult discrimination IC card "taspo" was introduced in 2008 to prevent minors from purchasing cigarettes in Japan. This study aimed to elucidate the short-term change in smoking behavior among a cohort of high school students through the introduction of the taspo system. We conducted a questionnaire survey in students at one high school in the metropolitan area of Japan in 2008. In this area, the taspo system was introduced on July 1, and the survey was conducted before and after its introduction (June and September). Change in smoking behavior was examined by linking the two questionnaires using a unique identification number for each participant. The questionnaire included basic characteristics, smoking-related behavior, and means of obtaining tobacco. Of 133 students, 123 (response rate 84.7%) completed the before and after questionnaire forms and could be linked. The smoking rate was 22.8% in June and 25.2% in September, with no statistically significant change. Vending machines were the major means of obtaining tobacco in June, while the use of cigarette shops and supermarkets increased after the introduction of taspo. The introduction of taspo hardly influenced underage smoking behavior during the observation period in our study subjects. The only significant change was in the means of obtaining tobacco. To prevent underage smoking, the importance of comprehensive restriction of the procurement route was suggested.


[How to find a place in the world: exploring people's experiences recovering from serious mental disorders].
This historical, conceptual, and cultural review of the concept of recovery within the realm of mental health discusses the last decade's research into the subjective experiences of people in the process of recovering from serious mental disorders and analyzes the differences between the notions of rehabilitation, integration, and recovery. After presenting a historical introduction, the article describes recovery experiences from three angles: identity and control over one's own life; personal and community relationships; and material resources, work, and other social activities. The concept of recovery entails new challenges and tensions that must be taken into account in the reform of mental health systems in Europe and the Americas.


Transcranial Doppler pulsatility index: not an accurate method to assess intracranial pressure.
Transcranial Doppler sonography (TCD) assessment of intracranial blood flow velocity has been suggested to accurately determine intracranial pressure (ICP). We attempted to validate this method in patients with communicating cerebrospinal fluid systems using predetermined pressure levels. Ten patients underwent a lumbar infusion test, applying 4 to 5 preset ICP levels. On each level, the pulsatility index (PI) in the middle cerebral artery was determined by measuring the blood flow velocity using TCD. ICP was simultaneously measured with an intraparenchymal sensor. ICP and PI were compared using correlation analysis. For further understanding of the ICP-PI relationship, a mathematical model of the intracranial dynamics was simulated using a computer. The ICP-PI regression equation was based on data from 8 patients. For 2 patients, no audible Doppler signal was obtained. The equation was ICP = 23*PI + 14 (R = 0.22, P < .01, N = 35). The 95% confidence interval for a mean ICP of 20 mm Hg was -3.8 to 43.8 mm Hg. Individually, the regression coefficients varied from 42 to 90 and the offsets from -32 to +3. The mathematical simulations suggest that variations in vessel compliance, autoregulation, and arterial pressure have a serious effect on the ICP-PI relationship. The in vivo results show that PI is not a reliable predictor of ICP. Mathematical simulations indicate that this is caused by variations in physiological parameters.


Arcobacter butzleri is an opportunistic pathogen: recurrent bacteraemia in an immunocompromised patient without diarrhoea.
Arcobacter butzleri is attracting increasing interest due to its possible pathogenic properties. Researchers have described cases in which A. butzleri is isolated in stool samples from patients with gastrointestinal symptoms, mostly diarrhoea. The relevance of adding our case to the literature lies in its description of recurrent A. butzleri bacteraemia in a patient without diarrhoea. An immunocompromised patient was hospitalized three times within 12 months due to A. butzleri-induced bacteraemia. At no time did the patient experience diarrhoea even though examination of stool samples showed growth of A. butzleri . The isolate was susceptible to gentamicin, colistin and tetracyclines. The patient was successfully treated with doxycycline. For the first time in the literature we describe recurrent A. butzleri bacteraemia in a patient without diarrhoea. This case supports the classification of A. butzleri as an opportunistic pathogenic species, which clinical microbiology laboratories should be able to identify.


HDAC6: A Key Link Between Mitochondria and Development of Peripheral Neuropathy.
Peripheral neuropathy, which is the result of nerve damage from lesions or disease, continues to be a major health concern due to the common manifestation of neuropathic pain. Most investigations into the development of peripheral neuropathy focus on key players such as voltage-gated ion channels or glutamate receptors. However, emerging evidence points to mitochondrial dysfunction as a major player in the development of peripheral neuropathy and resulting neuropathic pain. Mitochondrial dysfunction in neuropathy includes altered mitochondrial transport, mitochondrial metabolism, as well as mitochondrial dynamics. The mechanisms that lead to mitochondrial dysfunction in peripheral neuropathy are poorly understood, however, the Class IIb histone deacetylase (HDAC6), may play an important role in the process. HDAC6 is a key regulator in multiple mechanisms of mitochondrial dynamics and may contribute to mitochondrial dysregulation in peripheral neuropathy. Accumulating evidence shows that HDAC6 inhibition is strongly associated with alleviating peripheral neuropathy and neuropathic pain, as well as mitochondrial dysfunction, in in vivo and in vitro models of peripheral neuropathy. Thus, HDAC6 inhibitors are being investigated as potential therapies for multiple peripheral neuropathic disorders. Here, we review emerging studies and integrate recent advances in understanding the unique connection between peripheral neuropathy and mitochondrial dysfunction through HDAC6-mediated interactions.


Impact of environmental and genetic factors on biofilm formation by the probiotic strain Lactobacillus rhamnosus GG.
Lactobacillus rhamnosus GG (ATCC 53103) is one of the clinically best-studied probiotic organisms. Moreover, L. rhamnosus GG displays very good in vitro adherence to epithelial cells and mucus. Here, we report that L. rhamnosus GG is able to form biofilms on abiotic surfaces, in contrast to other strains of the Lactobacillus casei group tested under the same conditions. Microtiter plate biofilm assays indicated that in vitro biofilm formation by L. rhamnosus GG is strongly modulated by culture medium factors and conditions related to the gastrointestinal environment, including low pH; high osmolarity; and the presence of bile, mucins, and nondigestible polysaccharides. Additionally, phenotypic analysis of mutants affected in exopolysaccharides (wzb), lipoteichoic acid (dltD), and central metabolism (luxS) showed their relative importance in biofilm formation by L. rhamnosus GG.


Metabolic and physiologic responses to video game play in 7- to 10-year-old boys.
To examine the metabolic, physiologic, and hemostatic responses to action video game play in a group of young boys. Comparison study. Laboratory of Clinical and Applied Physiology, University of Miami. Twenty-one boys aged 7 to 10 years. Blood pressure monitored before and during game play and blood glucose and lactate levels measured before and immediately after game play. Measurements were continuously recorded throughout game play. Dependent t tests were used to compare measurements recorded at baseline and during or after game play. Effect sizes using the Cohen d were examined for comparisons. Significant increases from baseline were found for heart rate (18.8%; P<.001), systolic (22.3%; P<.001) and diastolic (5.8%; P = .006) blood pressure, ventilation (51.9%; P<.001), respiratory rate (54.8%; P<.001), oxygen consumption (49.0%; P<.001), and energy expenditure (52.9%; P<.001). Effect sizes for these comparisons were medium or large. No significant changes were found from baseline to after video game play for lactate (18.2% increase; P = .07) and glucose (0.9% decrease; P = .59) levels. Video game play results in significant increases in various metabolic and physiologic variables in young children. Thus, it should not be combined with television viewing for the evaluation of sedentary activities. The magnitude of change, however, was lower than that observed during standard physical exercise and below national health recommendations. As such, video game play should not be considered a substitute for regular physical activities that significantly stress the metabolic pathways required for the promotion of cardiovascular conditioning.


Motion planning and coordination for robot systems based on representation space.
This paper proposes a general motion planning and coordination strategy for robot systems. The representation space (RS) of a robot system is constructed to describe the distributions of system attributes. The reachable area in the RS, denoting the attribute set that the system can be of, indicates the system's ability to accomplish tasks. Moreover, it also describes the influences of the internal and external constraints on the system's capability. Task realization is transformed to finding a trajectory in the RS for the system attributes to transit along under constraints. Meanwhile, the realizable conditions of a prescribed task by the robot system of specific configurations are discussed. If the task is realizable, the optimal strategy for task execution could further be figured out. Otherwise, it could be transformed to be realizable via task reassignment or system reconfigurations so that a connected path could be found for the transition of the system attributes from the starting point to the goal in the RS. The proposed scheme contributes to designing, planning, and coordination of the robotic tasks. Experiments on path planning of a robot manipulator and formation movement of a multirobot system, as well as coordination of a mobile manipulator system, are conducted to show the validity and generalization of the proposed method.


Prevalence of metastases in hepatocellular carcinoma: risk factors and impact on survival.
The purpose of this study was to determine the prevalence and risk factors of metastases in hepatocellular carcinoma (HCC) patients and analyze the effects of different locations of metastases on survival. Retrospective analysis was performed on 347 HCC patients who received a metastatic workup including bone scan and computed tomography scans of chest, abdomen, and pelvis. Clinical and tumor characteristics were evaluated as risk factors for metastasis by univariate and multivariate methods. Survival was analyzed by Kaplan-Meier and Cox regression methods. One hundred forty-five patients had metastases: 72 had thoracic, 57 had abdominal, and 34 had bone metastases. Significant differences were noted with weight loss, hepatitis C, tumor grade, tumor multifocality, size, and alkaline phosphatase levels between the metastases group and the nonmetastases group by univariate analysis. Poor differentiation, multilobar spread, and size (> or = 5 cm) were strongest predictors of metastatic disease by logistic regression. Patients with thoracic metastases had significantly poorer survival. HCC metastasis is prevalent on initial presentation. Evaluation for liver transplantation or curative resection requires a full metastatic workup. Poor differentiation, larger tumors, and those with multilobar spread have increased risk for metastasis. Patients with thoracic spread have poor prognosis as compared to other locations of metastasis.


Discovery of a new dihydroeugenol-chalcone hybrid with cytotoxic and anti-migratory potential: A dual-action hit for cancer therapeutics.
Cancer still represents a serious public health problem and one of the main problems related to the worsening of this disease is the ability of some tumors to develop metastasis. In this work, we synthesized a new series of chalcones and isoxazoles derived from eugenol and analogues as molecular hybrids and these compounds were evaluated against different tumor cell lines. This structural pattern was designed considering the cytotoxic potential already known for eugenol, chalcones and isoxazoles. Notably, chalcones 7, 9, 10, and 11 displayed significant activity (4.2-14.5 µM) against two cancer cell lines, surpassing the potency of the control drug doxorubicin. The reaction of chalcones with hydroxylamine hydrochloride provided the corresponding isoxazoles that were inactive against these cancer cells. The dihydroeugenol chalcone 7 showed the most promising results, demonstrating higher potency against HepG2 (CC50: 4.2 µM) and TOV-21G (CC50: 7.2 µM). Chalcone 7 was also three times less toxic than doxorubicin considering HepG2 cells, with a selectivity index greater than 11. Further investigations including clonogenic survival, cell cycle progression and cell migration assays confirmed the compelling antitumoral potential of chalcone 7, as it reduced long-term survival due to DNA fragmentation, inducing cell death and inhibiting HepG2 cells migration. Moreover, in silico studies involving docking and molecular dynamics revealed a consistent binding mode of chalcone 7 with metalloproteinases, particularly MMP-9, shedding light on its potential mechanism of action related to anti-migratory effects. These significant findings suggest the inclusion of compound 7 as a promising candidate for future studies in the field of cancer therapeutics.


Material properties of the dentate maxilla.
The aim of this study was to determine regional variability of material properties in the dentate maxilla. Cortical samples were removed from 15 sites of 15 adult dentate fresh-frozen maxillas. Cortical thickness, density, elastic properties, and the direction of greatest stiffness were obtained. Results showed that cortical bone in the alveolar region tended to be thicker, less dense, and less stiff. Cortical bone from the body of the maxilla was thinner, denser, and stiffer. Palatal cortical bone was intermediate in some features but overall was more similar to cortical bone from the alveolar region. The principal axes of stiffness varied regionally. The regions with the greatest consistency were the alveolar area and the frontomaxillary pillar, where the grain of the cortical bone was aligned vertically from the incisors to the medial external aspect of the orbit. Elastic properties in the human maxilla, especially the orientation of the principal axes of stiffness, were more variable than in the mandible. Incorporation of these properties into finite-element models should improve their accuracy and reliability.


Gomesin peptides prevent proliferation and lead to the cell death of devil facial tumour disease cells.
The Tasmanian devil faces extinction due to devil facial tumour disease (DFTD), a highly transmittable clonal form of cancer without available treatment. In this study, we report the cell-autonomous antiproliferative and cytotoxic activities exhibited by the spider peptide gomesin (AgGom) and gomesin-like homologue (HiGom) in DFTD cells. Mechanistically, both peptides caused a significant reduction at G0/G1 phase, in correlation with an augmented expression of the cell cycle inhibitory proteins p53, p27, p21, necrosis, exacerbated generation of reactive oxygen species and diminished mitochondrial membrane potential, all hallmarks of cellular stress. The screening of a novel panel of AgGom-analogues revealed that, unlike changes in the hydrophobicity and electrostatic surface, the cytotoxic potential of the gomesin analogues in DFTD cells lies on specific arginine substitutions in the eight and nine positions and alanine replacement in three, five and 12 positions. In conclusion, the evidence supports gomesin as a potential antiproliferative compound against DFTD disease.


A MultiTEP platform-based epitope vaccine targeting the phosphatase activating domain (PAD) of tau: therapeutic efficacy in PS19 mice.
Pathological tau correlates well with cognitive impairments in Alzheimer's disease (AD) patients and therefore represents a promising target for immunotherapy. Targeting an appropriate B cell epitope in pathological tau could in theory produce an effective reduction of pathology without disrupting the function of normal native tau. Recent data demonstrate that the N-terminal region of tau (aa 2-18), termed the "phosphatase activation domain (PAD)", is hidden within native Tau in a 'paperclip'-like conformation. Conversely, PAD is exposed in pathological tau and plays an essential role in the inhibition of fast axonal transport and tau polymerization. Thus, we hypothesized that anti-tau2-18 antibodies may safely and specifically reduce pathological tau and prevent further aggregation, which in turn would neutralize tau toxicity. Therefore, we evaluated the immunogenicity and therapeutic efficacy of our MultiTEP platform-based vaccine targeting tau2-18 formulated with AdvaxCpG adjuvant (AV-1980R/A) in PS19 tau transgenic mice. The AV-1980R/A induced extremely high antibody responses and the resulting sera recognized neurofibrillary tangles and plaque-associated dystrophic neurites in AD brain sections. In addition, under non-denaturing conditions AV-1980R/A sera preferentially recognized AD-associated tau. Importantly, vaccination also prevented age-related motor and cognitive deficits in PS19 mice and significantly reduced insoluble total and phosphorylated tau species. Taken together, these findings suggest that predominantly targeting misfolded tau with AV-1980R/A could represent an effective strategy for AD immunotherapy.


Racial variations in the incidence of congenital malformations.
Racial variation in the incidence of several common congenital malformations has been studied in Atlanta, Georgia. The white-to-black incidence ratios were substantially different from unity for single defects, while approximate equality of rates was noted for multiple defects. Amongst the specific malformations for which familial data were available, affected first- and second-degree relatives were found only in the families of probands with single defects. It is postulated that these findings may indicate a more prominent function of the environment in the genesis of multiple malformations than is the case for single defects, the occurrence of which may be more strongly affected by non-sporadic genetic factors.


Ultracryotomy of biological tissues to preserve membrane structure.
A vacuum transfer stage is described which permits visualization of ultracryotome sections without the considerable distortions found in sections which have either thawed or rehydrated after freeze drying. Membrane structure-nucleus, nucleolus, mitochondria with their cristae and plasma membrane-was observed only in cells at the surface of the tissue which had undergone the fastest freezing rates. Inspection of knife damage in sections through these superficial cells showed that glass or diamond knives which are perfectly adequate for sectioning resin-embedded tissues are less suited to sectioning frozen biological tissue. Deeper in the tissue, where the freezing rates were slower, ice crystal cavities destroyed all membranous structures.


Experience with the Strata valve in the management of shunt overdrainage.
The overdrainage of cerebrospinal fluid (CSF) in children with shunt-treated hydrocephalus may cause chronic disabling symptoms and require repeated surgery. Externally adjustable valves offer a noninvasive way of altering the valve opening pressure. The authors report on their experience with using the Strata valve in the management of symptomatic CSF overdrainage. The authors treated 24 patients with symptomatic CSF overdrainage by inserting a Strata valve. The severity of symptoms was graded, and the frequency of hospital visits and shunt operations was recorded before and after insertion of the valve. Additionally, results of brain imaging and intracranial pressure monitoring were reviewed. Nineteen patients (79.2%) had severe symptoms at the time of the insertion; 1 year after Strata valve insertion only one patient (4.17%) still suffered severe symptoms. The number of hospital admissions was 3.38/patient/year before placement and 1.21 for the 1st year, 1 for the 2nd, and 0.4 for the 3rd postoperative year. The number of operations was 3.42/patient/year during the year before placement of the valve, and then 0.71 for the 1st, 0.56 for the 2nd, and 0.25 for the 3rd postoperative years. During the 1st year after placement of the Strata valve, the settings were changed 2.79 times/patient/year, 1.29 for the 2nd, and 1.33 times/patient/year for the 3rd year. The Strata valve was effective in improving the symptoms of overdrainage in the majority of patients in this series. The number of hospital admissions and operations for valve malfunction was reduced.


Pancreatic cancer and autoimmune diseases: An association sustained by computational and epidemiological case-control approaches.
Deciphering the underlying genetic basis behind pancreatic cancer (PC) and its associated multimorbidities will enhance our knowledge toward PC control. The study investigated the common genetic background of PC and different morbidities through a computational approach and further evaluated the less explored association between PC and autoimmune diseases (AIDs) through an epidemiological analysis. Gene-disease associations (GDAs) of 26 morbidities of interest and PC were obtained using the DisGeNET public discovery platform. The association between AIDs and PC pointed by the computational analysis was confirmed through multivariable logistic regression models in the PanGen European case-control study population of 1,705 PC cases and 1,084 controls. Fifteen morbidities shared at least one gene with PC in the DisGeNET database. Based on common genes, several AIDs were genetically associated with PC pointing to a potential link between them. An epidemiologic analysis confirmed that having any of the nine AIDs studied was significantly associated with a reduced risk of PC (Odds Ratio (OR) = 0.74, 95% confidence interval (CI) 0.58-0.93) which decreased in subjects having ≥2 AIDs (OR = 0.39, 95%CI 0.21-0.73). In independent analyses, polymyalgia rheumatica, and rheumatoid arthritis were significantly associated with low PC risk (OR = 0.40, 95%CI 0.19-0.89, and OR = 0.73, 95%CI 0.53-1.00, respectively). Several inflammatory-related morbidities shared a common genetic component with PC based on public databases. These molecular links could shed light into the molecular mechanisms underlying PC development and simultaneously generate novel hypotheses. In our study, we report sound findings pointing to an association between AIDs and a reduced risk of PC.


[The functional state of platelets in kidney cancer].
Dynamic function of thrombocytes was studied in 75 cases of renal cancer. Enhanced thrombocytic hemostasis registered in patients with stage I-II tumor was matched by a low-level thrombocytic function with concomitant thrombocythemia at stages III-IV. These findings may be useful in prevention and treatment of thrombohemorrhagic complications in renal cancer.


Uncovering the Properties of Dicationic Ionic Liquid Nanodroplets through Ab Initio Molecular Dynamics Simulations.
The behavior of nanodroplets of an imidazolium-based dicationic ionic liquid, i.e., [C1(mim)2][PF6]2, was investigated in this study using ab initio molecular dynamics simulations. The vibrational features as well as the structural, interfacial, and dynamical properties of different sized droplets were analyzed and compared to the bulk phase system. Structural properties of the droplets, such as π-π stacking, radial distribution functions, structure factors, combined distribution functions, and angular distribution functions were analyzed to understand the interactions and orientations of their ions. The vibrational features and hydrogen bonding strength of droplets were studied by calculating their infrared (IR) and power spectra, determining the contribution of different types of hydrogen bonding to each vibrational mode. The calculated spectra showed good overall agreement with the experimental results. The interfacial properties of the droplets and the orientation of their ions were analyzed using density profiles and an exposed surface. The results showed that, in all systems studied, cations and anions were equally likely to exist in both inner and outer layers, and the cations tended to be oriented toward the center of droplets with obtuse angles. Additionally, the droplet densities were extrapolated to predict the bulk phase density with less than 2% deviation. The dynamical properties of hydrogen bonds, mean square displacement, and van Hove correlations of cations and anions were also analyzed. The results indicated that there was no regular trend in the dynamic properties of droplets with an increasing system size.


Intersectoriality in Danish municipalities: corrupting the social determinants of health?
Action on the social determinants of health (SDH) through intersectoral policymaking is often suggested to promote health and health equity. This paper argues that the process of intersectoral policymaking influences how the SDH are construed and acted upon in municipal policymaking. We discuss how the intersectoral policy process legitimates certain practices in the setting of Danish municipal health promotion and the potential impact this can have for long-term, sustainable healthy public policy. Based on ethnographic fieldwork, we show how the intention of intersectoriality produces a strong concern for integrating health into non-health sectors to ensure productive collaboration. To encourage this integration, health is often framed as a means to achieve the objectives of non-health sectors. In doing so, the intersectoral policy process tends to favor smaller-scale interventions that aim to introduce healthier practices into various settings, e.g. creating healthy school environments for increased physical activity and healthy eating. While other more overarching interventions on the health impacts of broader welfare policies (e.g. education policy) tend to be neglected. The interventions hereby neglect to address more fundamental SDH. Based on these findings, we argue that intersectoral policymaking to address the SDH may translate into a limited approach to action on so-called 'intermediary determinants' of health, and as such may end up corrupting the broader SDH. Further, we discuss how this corruption affects the intended role of non-health sectors in tackling the SDH, as it may impede the overall success and long-term sustainability of intersectoral efforts.


Pulmonary arteriovenous fistula manifesting as amaurosis fugax--case report.
Pulmonary arteriovenous fistula (PAVF) is a rare condition which occasionally causes neurological complications. A 43-year-old female with multiple PAVFs presented with several episodes of amaurosis fugax and transient right hemiparesis. She had no other vascular abnormality, and her human leukocyte antigen haplotype did not coincide with previous patients with hereditary hemorrhagic telangiectasia. She underwent PAVF embolization to prevent further neurological complications, and had an uneventful subsequent clinical course. Amaurosis fugax is a slight neurological symptom and may be an early important sign of PAVF. We stress that PAVFs should be considered in the differential diagnosis of patients with amaurosis fugax who complain of exertional dyspnea or demonstrate cyanosis.


Diabetes and healing: a review of the literature.
Blood sugar control is an important factor for healing in the diabetic surgical patient. Careful assessment preoperatively of all patients for glucose is essential. A surgical zone of serum glucose should be established for diabetics, and this zone should be maintained throughout the procedure and postoperative period. Blood sugar control should be maintained by the patient throughout the healing phase, as poor control may interfere with the healing process.


Kock continent ileostomy in Indian patients.
Reservoir (continent) ileostomy was performed in 16 of our cases who had undergone total proctocolectomy. The basic pathology in these cases was chronic ulcerative colitis. Four cases (25%) had to be re-operated for partial or complete extrusion of the nipple valve causing incontinence. Two of these cases needed a second revision. By improving the valve formation technique, the incidence of sliding of the exit-valve was reduced. Other postoperative complications also responded well to this treatment. The patients were followed for three to five years. Three cases need frequent evacuation, but the remaining 13 cases, including some who had undergone revisional surgery, are fully continent. This procedure is accepted well by our patients.


Stereoselective metabolism of 7-chlorobenz(a)anthracene by rat liver microsomes. Absolute configurations and optical purities of trans-dihydrodiol metabolites.
Metabolism of 7-chlorobenz(a)anthracene (7-Cl-BA) by liver microsomes from untreated rats, and from rats pretreated with either 3-methylcholanthrene or phenobarbital was studied. The metabolites were isolated by HPLC and characterized by UV-visible, mass, and proton NMR spectral analyses and identified as 7-Cl-BA trans-3,4-, 5,6-, and 8,9-dihydrodiols, 3-hydroxy-7-Cl-BA, and 4-hydroxy-7-Cl-BA. Proton NMR spectral analysis indicated that 7-Cl-BA trans-3,4-dihydrodiol preferentially adopted a quasidiequatorial conformation while trans-5,6- and 8,9-dihydrodiols preferentially adopted a quasidiaxial conformation. Comparison of circular dichroism spectra with those of 7-bromobenz(a)anthracene trans-dihydrodiol metabolites of known absolute stereochemistry indicated that the major enantiomeric 7-Cl-BA trans-3,4-, 5,6-, and 8,9-dihydrodiols had R,R absolute configuration. Application of chiral stationary phase HPLC for direct resolution of the trans-dihydrodiols and their hydrogenated and dechlorinated derivatives enabled determination of the optical purity of each dihydrodiol metabolite obtained from the three microsomal systems. In vitro incubation of 7-Cl-BA under molecular oxygen-18 produced 7-Cl-BA trans-3,4-, 5,6-, and 8,9-dihydrodiols, each containing one oxygen-18 atom. Mass spectral analysis of the dehydration products of the oxygen-18-containing trans-dihydrodiol metabolites indicated that 7-Cl-BA 3S,4R-epoxide and 7-Cl-BA 8R,9S-epoxide were the predominant enantiomeric intermediates.


Three dimensional visualization of the statically equivalent serial chain from kinect recording.
We develop and present a portable tool intended for real time estimation of the center of mass (CoM) in human subjects. Using the statically equivalent serial chain (SESC) method we can account for subject specificity after identification of the model's parameters. CoM position estimates are then available from measurements of the subject's limbs orientations. For portability, we make use of widely accessible sensors such as the Kinect and Wii balance board for identification. Use of the Kinect as a measurement device allows us to establish the SESC outside of the laboratory, without many special considerations on the environment. Only Kinect is used for CoM tracking after identification was performed. We present here an overview of the SESC concept and the identification procedure. The aspects involved in the visualization tool are discussed and results are shown in order to verify the performance.


Context-specific activation of hippocampus and SN/VTA by reward is related to enhanced long-term memory for embedded objects.
Animal studies indicate that hippocampal representations of environmental context modulate reward-related processing in the substantia nigra and ventral tegmental area (SN/VTA), a major origin of dopamine in the brain. Using functional magnetic resonance imaging (fMRI) in humans, we investigated the neural specificity of context-reward associations under conditions where the presence of perceptually similar neutral contexts imposed high demands on a putative hippocampal function, pattern separation. The design also allowed us to investigate how contextual reward enhances long-term memory for embedded neutral objects. SN/VTA activity underpinned specific context-reward associations in the face of perceptual similarity. A reward-related enhancement of long-term memory was restricted to the condition where the rewarding and the neutral contexts were perceptually similar, and in turn was linked to co-activation of the hippocampus (subfield DG/CA3) and SN/VTA. Thus, an ability of contextual reward to enhance memory for focal objects is closely linked to context-related engagement of hippocampal-SN/VTA circuitry.


The atrial and ventricular myocardial proteome of end-stage lamin heart disease.
Lamins A/C (encoded by LMNA gene) can lead to dilated cardiomyopathy (DCM). This pilot study sought to explore the postgenomic phenotype of end-stage lamin heart disease. Consecutive patients with end-stage lamin heart disease (LMNA-group, n = 7) and ischaemic DCM (ICM-group, n = 7) undergoing heart transplantation were prospectively enrolled. Samples were obtained from left atrium (LA), left ventricle (LV), right atrium (RA), right ventricle (RV) and interventricular septum (IVS), avoiding the infarcted myocardial segments in the ICM-group. Samples were analysed using a discovery 'shotgun' proteomics approach. We found that 990 proteins were differentially abundant between LMNA and ICM samples with the LA being most perturbed (16-fold more than the LV). Abundance of lamin A/C protein was reduced, but lamin B increased in LMNA LA/RA tissue compared to ICM, but not in LV/RV. Carbonic anhydrase 3 (CA3) was over-abundant across all LMNA tissue samples (LA, LV, RA, RV, and IVS) when compared to ICM. Transthyretin was more abundant in the LV/RV of LMNA compared to ICM, while sarcomeric proteins such as titin and cardiac alpha-cardiac myosin heavy chain were generally less abundant in RA/LA of LMNA. Protein expression profiling and enrichment analysis pointed towards sarcopenia, extracellular matrix remodeling, deficient myocardial energetics, redox imbalances, and abnormal calcium handling in LMNA samples. Compared to ICM, end-stage lamin heart disease is a biventricular but especially a biatrial disease appearing to have an abundance of lamin B, CA3 and transthyretin, potentially hinting to compensatory responses.


Fold-change threshold screening: a robust algorithm to unmask hidden gene expression patterns in noisy aggregated transcriptome data.
Transcriptomics is often used to investigate changes in an organism's genetic response to environmental contamination. Data noise can mask the effects of contaminants making it difficult to detect responding genes. Because the number of genes which are found differentially expressed in transcriptome data is often very large, algorithms are needed to reduce the number down to a few robust discriminative genes. We present an algorithm for aggregated analysis of transcriptome data which uses multiple fold-change thresholds (threshold screening) and p values from Bayesian generalized linear model in order to assess the robustness of a gene as a potential indicator for the treatments tested. The algorithm provides a robustness indicator (ROBI) as well as a significance profile, which can be used to assess the statistical significance of a given gene for different fold-change thresholds. Using ROBI, eight discriminative genes were identified from an exemplary dataset (Danio rerio FET treated with chlorpyrifos, methylmercury, and PCB) which could be potential indicators for a given substance. Significance profiles uncovered genetic effects and revealed appropriate fold-change thresholds for single genes or gene clusters. Fold-change threshold screening is a powerful tool for dimensionality reduction and feature selection in transcriptome data, as it effectively reduces the number of detected genes suitable for environmental monitoring. In addition, it is able to unmask patterns in altered genetic expression hidden by data noise and reduces the chance of type II errors, e.g., in environmental screening.


Identical clonal origin of synchronous and metachronous low-grade, noninvasive papillary transitional cell carcinomas of the urinary tract.
One of the most important biological features of papillary transitional cell carcinoma (pTCC) of the urinary tract is its multicentricity and its tendency for recurrences. Two possible mechanisms, field effect and intramucosal seeding/spreading, have been proposed. The former theory hypothesizes that carcinogenic agents cause synchronous or metachronous malignant transformation of multiple urothelial cells (independent clonal origin), and the latter speculates that synchronous and metachronous tumors are derived from implantation or direct spreading of tumor cells (identical clonal origin). We tested these hypotheses by analyzing the methylation patterns of the androgen receptor gene (HUMARA) located at the X-chromosome. Thirty-five metachronous and synchronous, low-grade (grade 1 or 2), noninvasive pTCCs of the urinary tract from 10 heterozygous female patients were successfully analyzed using formalin-fixed, paraffin-embedded tissue. These included 16 recurrent bladder tumors from 4 patients, 10 metachronous bladder and ureter/renal pelvis tumors from 4 patients, and 9 multifocal tumors from 2 patients. All tumors are monoclonal as indicated by unbalanced methylation of HUMARA. Furthermore, same methylated allele was detected in multiple recurrent or multifocal tumors from any given patient, indicating their identical clonal origin. We conclude that low-grade, noninvasive pTCCs are monoclonal in nature. Synchronous or metachronous pTCCs have an identical clonal origin, strongly supporting the intramucosal seeding/spreading hypothesis.


Spatial integration characteristics in motion detection and direction identification.
Spatial integration characteristics were assessed with drifting gratings for both detection and direction-identification contrast thresholds. Thresholds were measured while stimulus width, length or both were varied. It was found that: (1) the shape of the size/sensitivity functions changes with spatial, but not with temporal, frequency; (2) direction-identification thresholds diverge from the detection thresholds below 1 cycle but can be reliably measured for stimulus widths as small as 0.1275 cycles; (3) the integration characteristics are slightly anisotropic for the identification but not for the detection process, and (4) the two-dimensional spatial integration cannot be directly predicted from its one-dimensional characteristics. Width/sensitivity detection functions are well fitted by predictions of Wilson and Bergen's four-channel model. Predictions from a temporal covariance model provide a poor fit to the identification data. It is argued that classes of detection and direction-identification models must involve identical nonlinearities prior to their respective thresholds. It is concluded that the hypothesis according to which both performances are determined by the same spatial integration stage cannot be rejected.


Expression of human decay-accelerating factor on intestinal epithelium of transgenic mice does not facilitate infection by the enteral route.
In vitro, infection of polarized human intestinal epithelial cells by coxsackievirus B3 (CVB3) depends on virus interaction with decay-accelerating factor (DAF), a receptor expressed on the apical cell surface. Although mice are highly susceptible to CVB3 infection when virus is delivered by intraperitoneal injection, infection by the enteral route is very inefficient. Murine DAF, unlike human DAF, does not bind virus, and we hypothesized that the absence of an accessible receptor on the intestinal surface is an important barrier to infection by the oral route. We generated transgenic mice that express human DAF specifically on intestinal epithelium and measured their susceptibility to infection by a DAF-binding CVB3 isolate. Human DAF permitted CVB3 to bind to the intestinal surface ex vivo and to infect polarized monolayers of small-intestinal epithelial cells derived from DAF transgenic mice. However, expression of human DAF did not facilitate infection by the enteral route either in immunocompetent animals or in animals deficient in the interferon alpha/beta receptor. These results indicate that the absence of an apical receptor on intestinal epithelium is not the major barrier to infection of mice by the oral route. CVB3 infection of human intestinal epithelial cells depends on DAF at the apical cell surface, and expression of human DAF on murine intestinal epithelial cells permits their infection in vitro. However, expression of human DAF on the intestinal surface of transgenic mice did not facilitate infection by the oral route. Although the role of intestinal DAF in human infection has not been directly examined, these results suggest that DAF is not the critical factor in mice.


Sulfurihydrogenibium rodmanii sp. nov., a sulfur-oxidizing chemolithoautotroph from the Uzon Caldera, Kamchatka Peninsula, Russia, and emended description of the genus Sulfurihydrogenibium.
Four thermophilic, sulfur-oxidizing, chemolithoautotrophic strains with >99 % 16S rRNA gene sequence similarity were isolated from terrestrial hot springs in the Geyser Valley and the Uzon Caldera, Kamchatka, Russia. One strain, designated UZ3-5T, was characterized fully. Cells of UZ3-5T were Gram-negative, motile, slightly oval rods (about 0.7 microm wide and 1.0 microm long) with multiple polar flagella. All four strains were obligately microaerophilic chemolithoautotrophs and could use elemental sulfur or thiosulfate as electron donors and oxygen (1-14 %, v/v) as the electron acceptor. Strain UZ3-5T grew at temperatures between 55 and 80 degrees C (optimally at 75 degrees C; 1.1 h doubling time), at pH 5.0-7.2 (optimally at pH 6.0-6.3) and at 0-0.9 % NaCl (optimally in the absence of NaCl). The G+C content of the genomic DNA of strain UZ3-5T was 35 mol%. Phylogenetic analysis revealed that strain UZ3-5T was a member of the genus Sulfurihydrogenibium, its closest relative in culture being Sulfurihydrogenibium azorense Az-Fu1T (98.3 % 16S rRNA gene sequence similarity). On the basis of its physiological and molecular characteristics, strain UZ3-5T represents a novel species of the genus Sulfurihydrogenibium, for which the name Sulfurihydrogenibium rodmanii sp. nov. is proposed. The type strain is UZ3-5T (=OCM 900T =ATCC BAA-1536T =DSM 19533T).


Attenuation of DNA damage in the dermis and epidermis of the albino hairless mouse by chronic exposure to ultraviolet-A and -B radiation.
Mammalian skin is vulnerable to the photocarcinogenic and photoaging effects of solar UV radiation and defends itself using a variety of photoprotective responses including epidermal thickening, tanning and the induction of repair and antiradical systems. We treated Skh-1 albino hairless mice for 60 days with ultraviolet-A (UVA) or ultraviolet-B (UVB) radiation and measured the frequency of cyclobutane pyrimidine dimers and pyrimidine(6-4)pyrimidone photoproducts induced by a single acute sunburn dose of UVB at different stages of the chronic treatment. We found that both UVA and UVB exposure produced a photoprotective response in the dermis and epidermis and that the degree of photoproduct attenuation was dependent on dose, wavelength and the type of damage induced. Although epidermal thickening was important, our data suggest that UV protective compounds other than melanin may be involved in mitigating the damaging effects of sunlight in the skin.


Easily Regenerated Readily Deployable Absorbent for Heavy Metal Removal from Contaminated Water.
Although clean and abundant water is the keystone of thriving communities, increasing demand and volatile climate patterns are depleting rivers and aquifers. Moreover, the quality of such water sources is threatened by noxious contaminants, of which heavy metals represents an area of growing concern. Recently, graphene oxide (GO) has been suggested as an adsorbent; however, a support is desirable to ensure a high surface area and an immobile phase. Herein, we described the preparation and characterization of a supported-epoxidized carbon nanotube (SENT) via the growth of multi walled carbon nanotubes (MWNTs) onto a quartz substrate. Subsequent epoxidation provides sufficient functionality to enable adsorbent of heavy metals (Cd2+, Co2+, Cu2+, Hg2+, Ni2+, and Pb2+) from aqueous solution with initial concentrations (60-6000 ppm) chosen to simulate high industrial wastewater contamination. The SENT adsorption efficiency is >99.4% for all metals and the saturation concentration is significantly greater than observed for either GO or acid treated MWNTs. The SENT adsorbent may be readily regenerated under mild conditions using a globally available household chemical, vinegar. 1 g of SENT has the potential to treat 83,000 L of contaminated water down to WHO limits which would be sufficient for 11,000 people.


Neurological outcomes in children with and without amalgam-related mercury exposure: seven years of longitudinal observations in a randomized trial.
Although large-scale, randomized trials involving children have been completed and their results demonstrate an absence of neurobehavioral effects from clinical exposure to mercury amalgam, neurological findings from such studies have not been reported. The authors conducted a randomized, prospective trial examining the safety of dental amalgam in which 507 children aged 8 through 12 years were assigned to treatment with either amalgam or resin-based composite. During seven years of follow-up, the authors performed annual clinical neurological examinations, including an evaluation of neurological hard signs (NHSs), presence of tremor and neurological soft signs (NSSs). The authors found no significant differences between treatment groups in any of the neurological measures. Groups did not differ with respect to the presence or absence of NHSs or tremor, nor the presence or absence or severity of NSSs at any point. As expected, NSS severity scores diminished with increasing age. Even at the levels of amalgam exposure in this study (a mean of 7.7-10.7 amalgam surfaces per subject across the seven years of follow-up), the authors conclude that exposure to mercury from dental amalgam does not adversely affect neurological status. The current evidence is that potential neurobehavioral or neurological effects from dental amalgam mercury exposure in children are inconsequential.


Impact of fill-level in twin-screw granulation on critical quality attributes of granules and tablets.
In a previous study a change of the fill-level in the barrel exerted a huge influence on the twin-screw granulation (TSG) process of a high drug loaded, simplified formulation. The present work investigated this influence systematically. The specific feed load (SFL) indicating the mass per revolution as surrogate parameter for the fill-level was applied and the correlation to the real volumetric fill level of an extruder could be demonstrated by a newly developed method. A design of experiments was conducted to examine the combined influence of SFL and screw speed on the process and on critical quality attributes of granules and tablets. The same formulation was granulated at constant liquid level with the same screw configuration and led to distinctively different results by only changing the fill-level and the screw speed. The power consumption of the extruder increased at higher SFLs with hardly any influence of screw speed. At low SFL the median residence time was mainly fill-level dependent and at higher SFL mainly screw speed dependent. Optimal values for the product characteristics were found at medium values for the SFL. Granule size distributions shifted from mono-modal and narrow shape to broader and even bimodal distributions of larger median granule sizes, when exceeding or falling below a certain fill-level. Deviating from the optimum fill-level, tensile strength of tablets decreased by about 25% and disintegration times of tablets increased for more than one third. At low fill-levels, material accumulation in front of the kneading zone was detected by pressure measurements and was assumed to be responsible for the unfavored product performance. At high fill-levels, granule consolidation due to higher propensity of contact with the result of higher material temperature was accounted for inferior product performance. The fill-level was found to be an important factor in assessment and development of twin-screw granulation processes as it impacted process and product attributes enormously.


Efficacy and safety of bempedoic acid among patients with and without diabetes: prespecified analysis of the CLEAR Outcomes randomised trial.
Statins reduce LDL cholesterol and cardiovascular events among those with or without diabetes but have been reported to increase new-onset diabetes. The CLEAR Outcomes trial demonstrated that bempedoic acid reduced the risk of major adverse cardiovascular events among statin-intolerant patients at high cardiovascular risk. In this prespecified analysis, our dual aims were to evaluate the cardiovascular benefits of bempedoic acid, an ATP-citrate lyase inhibitor, in individuals with diabetes, and to evaluate the risk of new-onset diabetes and HbA1c among those without diabetes in the CLEAR Outcomes trial. CLEAR Outcomes was a randomised, double-blind, placebo-controlled trial conducted across 1250 primary care and outpatient sites in 32 countries. Patients with or without cardiovascular disease who were unwilling or unable to take guideline-recommended doses of statins and an LDL cholesterol of 2·59 mmol/L or more were randomly assigned (1:1) in a double-blinded manner to either bempedoic acid 180 mg once per day or placebo. In this prespecified analysis, the efficacy endpoint was a time-to-event analysis of four-component major adverse cardiovascular event (MACE-4), which is the composite of cardiovascular death, nonfatal myocardial infarction, nonfatal stroke, or coronary revascularisation, using the intention-to-treat population stratified by baseline glycaemia status. The prespecified analysis of risk of new-onset diabetes and HbA1c increase was evaluated in patients without diabetes at baseline. The CLEAR Outcomes trial was completed on Nov 7, 2022, and is registered with ClinicalTrials.gov (NCT02993406). Between Dec 22, 2016, and Nov 7, 2022, 13 970 patients were screened and randomly assigned; 6373 (45·6%) with diabetes, 5796 (41·5%) with prediabetes, and 1801 (12·9%) with normoglycaemia. Over a median of 3·4 years follow up, patients with diabetes had significant relative and absolute cardiovascular risk reductions in MACE-4 endpoints with bempedoic acid (HR 0·83; 95% CI 0·72-0·95; absolute risk reduction of 2·4%) compared to placebo, with no statistical evidence of effect modification across glycaemic strata (interaction p=0·42). The proportion of patients who developed new-onset diabetes were similar between the bempedoic acid and placebo groups, with 429 of 3848 (11·1%) with bempedoic acid versus 433 of 3749 (11·5%) with placebo (HR 0·95; 95% CI 0·83-1·09). HbA1c concentrations at month 12 and the end of the study were similar between randomised groups in patients who had prediabetes and normoglycaemia. Placebo-corrected LDL cholesterol concentrations and high-sensitivity C-reactive protein at 6 months were reduced in each glycaemic stratum (diabetes, prediabtes, and normoglycaemia) for patients randomly assigned to bempedoic acid (all p<0·001). Among patients with diabetes, bempedoic acid reduces LDL cholesterol and high-sensitivity C-reactive protein and risk of cardiovascular events. Patients without diabetes had no increase in new-onset diabetes or worsening HbA1c with bempedoic acid. The efficacy and cardiometabolic safety profile of bempedoic acid makes it a clinical option for those with and without diabetes. Esperion Therapeutics.


Cell surface heparan sulfate proteoglycans control adhesion and invasion of breast carcinoma cells.
Cell surface proteoglycans interact with numerous regulators of cell behavior through their glycosaminoglycan chains. The syndecan family of transmembrane proteoglycans are virtually ubiquitous cell surface receptors that are implicated in the progression of some tumors, including breast carcinoma. This may derive from their regulation of cell adhesion, but roles for specific syndecans are unresolved. The MDA-MB231 human breast carcinoma cell line was exposed to exogenous glycosaminoglycans and changes in cell behavior monitored by western blotting, immunocytochemistry, invasion and collagen degradation assays. Selected receptors including PAR-1 and syndecans were depleted by siRNA treatments to assess cell morphology and behavior. Immunohistochemistry for syndecan-2 and its interacting partner, caveolin-2 was performed on human breast tumor tissue arrays. Two-tailed paired t-test and one-way ANOVA with Tukey's post-hoc test were used in the analysis of data. MDA-MB231 cells were shown to be highly sensitive to exogenous heparan sulfate or heparin, promoting increased spreading, focal adhesion and adherens junction formation with concomitantly reduced invasion and matrix degradation. The molecular basis for this effect was revealed to have two components. First, thrombin inhibition contributed to enhanced cell adhesion and reduced invasion. Second, a specific loss of cell surface syndecan-2 was noted. The ensuing junction formation was dependent on syndecan-4, whose role in promoting actin cytoskeletal organization is known. Syndecan-2 interacts with, and may regulate, caveolin-2. Depletion of either molecule had the same adhesion-promoting influence, along with reduced invasion, confirming a role for this complex in maintaining the invasive phenotype of mammary carcinoma cells. Finally, both syndecan-2 and caveolin-2 were upregulated in tissue arrays from breast cancer patients compared to normal mammary tissue. Moreover their expression levels were correlated in triple negative breast cancers. Cell surface proteoglycans, notably syndecan-2, may be important regulators of breast carcinoma progression through regulation of cytoskeleton, cell adhesion and invasion.


Associations of birthweight and infant growth with body composition at age 15--the COMPASS study.
Size at birth and postnatal growth have been positively associated with obesity in adulthood. However, associations between postnatal growth and body composition later in life have rarely been studied. The overall purpose was to explore the associations between birthweight, weight gain during first year of life and height, weight, body mass index, fat free mass index (FFMI), fat mass index, % fat mass (FM) and waist circumference in adolescence. The COMPASS study is a population-based study of adolescents from a well-defined area in Stockholm County, Sweden. Birth characteristics and weight during childhood were collected from registers and child health centre records, and body composition at age 15 years was measured by bioelectric impedance by trained nurses. Complete data were available for 2453 adolescents. Associations between predictor and outcome variables were assessed with linear regression modelling. Birthweight was positively associated with all outcome variables, except for %FM among girls. FFMI increased by 0.49 kg/m(2)[95% CI 0.34, 0.63] (boys) and 0.25 kg/m(2)[0.12, 0.38] (girls) per 1 SD increase in birthweight. Increased weight gain in infancy showed strong, positive associations with all measures of body composition. FFMI increased by 0.73 kg/m(2)[0.60, 0.87] (boys) and 0.63 kg/m(2)[0.50, 0.76] (girls) per unit increase in weight z-score during first year of life. The effect of increased weight gain in infancy was not modified by birthweight. Birthweight and postnatal growth were both positively related to body composition in adolescence. Increased weight gain during the first year of life had stronger effect than prenatal growth, suggesting infancy to be a more critical period.


Molecular dynamics of a thermostable multicopper oxidase from Thermus thermophilus HB27: structural differences between the apo and holo forms.
Molecular dynamic (MD) simulations have been performed on Tth-MCO, a hyperthermophilic multicopper oxidase from thermus thermophilus HB27, in the apo as well as the holo form, with the aim of exploring the structural dynamic properties common to the two conformational states. According to structural comparison between this enzyme and other MCOs, the substrate in process to electron transfer in an outer-sphere event seems to transiently occupy a shallow and overall hydrophobic cavity near the Cu type 1 (T1Cu). The linker connecting the β-strands 21 and 24 of the second domain (loop (β21-β24)(D2)) has the same conformation in both states, forming a flexible lid at the entrance of the electron-transfer cavity. Loop (β21-β24)(D2) has been tentatively assigned a role occluding the access to the electron-transfer site. The dynamic of the loop (β21-β24)(D2) has been investigated by MD simulation, and results show that the structures of both species have the same secondary and tertiary structure during almost all the MD simulations. In the simulation, loop (β21-β24)(D2) of the holo form undergoes a higher mobility than in the apo form. In fact, loop (β21-β24)(D2) of the holo form experiences a conformational change which enables exposure to the electron-transfer site (open conformation), while in the apo form the opposite effect takes place (closed conformation). To confirm the hypothesis that the open conformation might facilitate the transient electron-donor molecule occupation of the site, the simulation was extended another 40 ns with the electron-donor molecule docked into the protein cavity. Upon electron-donor molecule stabilization, loops near the cavity reduce their mobility. These findings show that coordination between the copper and the protein might play an important role in the general mobility of the enzyme, and that the open conformation seems to be required for the electron transfer process to T1Cu.


Development of a Personalized Intestinal Fibrosis Model Using Human Intestinal Organoids Derived From Induced Pluripotent Stem Cells.
Intestinal fibrosis is a serious complication of Crohn's disease. Numerous cell types including intestinal epithelial and mesenchymal cells are implicated in this process, yet studies are hampered by the lack of personalized in vitro models. Human intestinal organoids (HIOs) derived from induced pluripotent stem cells (iPSCs) contain these cell types, and our goal was to determine the feasibility of utilizing these to develop a personalized intestinal fibrosis model. iPSCs from 2 control individuals and 2 very early onset inflammatory bowel disease patients with stricturing complications were obtained and directed to form HIOs. Purified populations of epithelial and mesenchymal cells were derived from HIOs, and both types were treated with the profibrogenic cytokine transforming growth factor β (TGFβ). Quantitative polymerase chain reaction and RNA sequencing analysis were used to assay their responses. In iPSC-derived mesenchymal cells, there was a significant increase in the expression of profibrotic genes (Col1a1, Col5a1, and TIMP1) in response to TGFβ. RNA sequencing analysis identified further profibrotic genes and demonstrated differential responses to this cytokine in each of the 4 lines. Increases in profibrotic gene expression (Col1a1, FN, TIMP1) along with genes associated with epithelial-mesenchymal transition (vimentin and N-cadherin) were observed in TGFβ -treated epithelial cells. We demonstrate the feasibility of utilizing iPSC-HIO technology to model intestinal fibrotic responses in vitro. This now permits the generation of near unlimited quantities of patient-specific cells that could be used to reveal cell- and environmental-specific mechanisms underpinning intestinal fibrosis.


Sella turcica atypical teratoid/rhabdoid tumor complicated with lung metastasis in an adult female.
Here we present the case of a 60-year-old woman with a rare sellar region atypical teratoid/rhabdoid tumor (AT/RT), complicated by lung metastasis and treated with neurosurgery, radiotherapy, and chemotherapy. The patient had recurrent headache associated with left cavernous sinus syndrome after a previous endonasal transsphenoidal resection for a presumptive pituitary macroadenoma. Pituitary magnetic resonance imaging showed a tumor regrowth in the original location with a haemorrhagic component involving the left cavernous sinus. A near complete transsphenoidal resection of the sellar mass was performed followed by 3 months of stereotactic radiotherapy. Because of a worsening of the general clinical conditions, respiratory failure, and asthenia, the patient underwent a contrast enhanced computer tomography of the whole body which showed the presence of lung metastasis. The histopathological diagnosis on samples from pituitary and lung tissues was AT/RT. The patient survived 30 months after diagnosis regardless chemotherapy. In the adult, the AT/RT should be considered as a possible rare, aggressive, and malignant neoplasm localized in the sella turcica.


Diagnostic utility of lymphocyte subset analysis in AIDS case finding.
Abnormalities of lymphocyte subsets, especially low absolute number of helper T cells, are characteristically present in acquired immune deficiency syndrome (AIDS). Similar abnormalities can be found in patients with persistent generalized lymphadenopathy (PGL) or AIDS-related complex (ARC) and, to a lesser degree, in asymptomatic people who have been exposed to human T-lymphotropic virus type III (HTLV-III). Nevertheless, there appears to be a widespread perception that lymphocyte subset analysis may be useful in AIDS case finding within high-risk groups. We evaluated the diagnostic utility of absolute number of helper T cells and ratio of helper to suppressor T cells in 33 patients with AIDS, 43 patients with PGL who had been referred for lymph node biopsy, 90 patients with PGL and 195 male homosexual controls. At conventional cutoff levels the tests did not appear to revise the probability of AIDS upward to any clinically significant degree when the pretest probability of AIDS was low. Lymphocyte subset analysis does not appear to be a cost-effective method of AIDS case finding in identified groups at risk in which the prevalence of AIDS is low.


Treatment of circadian rhythm sleep disorders with light.
The human circadian system is normally synchronised with the solar day, insuring that alertness and performance peak during daytime hours and consolidated sleep occurs during the night. In circadian rhythm sleep disorders, the pattern of sleep-wake is misaligned with the patient's circadian system or the external environment, resulting in insomnia, fatigue, and deterioration in performance. Appropriately-timed exposure to bright light can reset the timing of sleep and wake to the desired times, and improve sleep quality and daytime alertness. The efficacy of bright light therapy, however, is dependent on the time-of-day of the circadian cycle that the light is administered. In this article, we examine the physiological basis for bright light therapy, and we discuss the application of light in the treatment of circadian rhythm sleep disorders including advanced and delayed sleep-phase disorder, free-running disorder (nonentrained type), shiftwork disorder and jet lag disorder. We review the laboratory and field studies which have established bright light therapy as an effective treatment for sleep-wake and circadian misalignment, and we also provide guidelines for the appropriate timing and safe use of bright light therapy.


Zinc-imidazolate polymers (ZIPs) as a potential carrier to brain capillary endothelial cells.
Herein, we report the synthesis and characterization of nanospheres of a biodegradable zinc-imidazolate polymers (ZIPs) as a proof-of-concept delivery vehicle into human brain endothelial cells, the main component of the blood-brain barrier (BBB). The ZIP particles can readily encapsulate functional molecules such as fluorophores and inorganic nanoparticles at the point of synthesis producing stable colloidal dispersions. Our results show that these biodegradable particles are not cytotoxic, and are able to penetrate and release cargo species to human brain capillary endothelial cells in vitro thus exhibiting significant potential as a novel platform for brain targeting treatments.


Why Adolescents Attempt Suicide: A Qualitative Study of the Transition from Ideation to Action.
To ameliorate the public health problem of adolescent suicide, it is imperative to go beyond simply establishing which factors increase risk. Multiple factors often interact in such a way that escalates suicide risk, and some combinations may be particularly perilous. Therefore, it is essential to examine the sequence and interplay of these various contributors to understand how they interact to confer risk. To enhance our understanding of this process, we used qualitative in-depth interviews with 20 psychiatrically hospitalized adolescents who had recently made a suicide attempt to investigate the factors that contributed to their attempts. In this qualitative analysis we 1) identified 16 separate factors that contributed to their suicide attempts, and 2) focused on the sequence and interplay between these factors in order to better understand why adolescents attempt suicide, with specific attention to which factors facilitated the transition from suicide ideation to action. Findings demonstrated that the strain caused by historical, sociocultural, and interpersonal factors alone was not enough to result in adolescents attempting suicide. For all but two participants, it was the interaction of intrapersonal factors that appeared to distort cognitions and/or elevate emotions to the point where they became intolerable and suicide became a viable option. These intrapersonal factors appeared to be the catalyst in the process from suicide ideation to action. Our findings suggest the need for specific strategies that address cognitive distortions, emotion dysregulation, and feelings of invalidation and entrapment as potential targets for interventions and prevention practices with adolescents at risk for suicide.


Transport and Secretion of the Wnt3 Ligand by Motor Neuron-like Cells and Developing Motor Neurons.
The vertebrate neuromuscular junction (NMJ) is formed by a presynaptic motor nerve terminal and a postsynaptic muscle specialization. Cumulative evidence reveals that Wnt ligands secreted by the nerve terminal control crucial steps of NMJ synaptogenesis. For instance, the Wnt3 ligand is expressed by motor neurons at the time of NMJ formation and induces postsynaptic differentiation in recently formed muscle fibers. However, the behavior of presynaptic-derived Wnt ligands at the vertebrate NMJ has not been deeply analyzed. Here, we conducted overexpression experiments to study the expression, distribution, secretion, and function of Wnt3 by transfection of the motor neuron-like NSC-34 cell line and by in ovo electroporation of chick motor neurons. Our findings reveal that Wnt3 is transported along motor axons in vivo following a vesicular-like pattern and reaches the NMJ area. In vitro, we found that endogenous Wnt3 expression increases as the differentiation of NSC-34 cells proceeds. Although NSC-34 cells overexpressing Wnt3 do not modify their morphological differentiation towards a neuronal phenotype, they effectively induce acetylcholine receptor clustering on co-cultured myotubes. These findings support the notion that presynaptic Wnt3 is transported and secreted by motor neurons to induce postsynaptic differentiation in nascent NMJs.


Translations: effects of viewpoint, feature, and naming on identifying repeatedly copied drawings.
We explored the tension between bottom-up and top-down contributions to object recognition in a collaboration between a visual artist and a cognitive psychologist. Initial pictorial renderings of objects and animals from various viewpoints were iteratively copied, and a series of drawings that changed from highly concrete images into highly abstract images was produced. In drawing identification in which sets were shown in reverse order, participants were more accurate, more confident, and quicker to correctly identify the evolving image when it was originally displayed from a canonical viewpoint with all salient features present. In drawing identification in which images were shown in random order, more abstract images could be resolved as a result of previously identifying a more concrete iteration of the same drawing. The results raise issues about the influence of viewpoint and feature on the preservation of pictorial images and about the role of labelling in the interpretation of ambiguous stimuli. In addition, the study highlights a procedure in which visual stimuli can degrade without necessitating a substantial loss of complexity.


Near total calcanectomy with rotational flap closure of large decubitus heel ulcerations complicated by calcaneal osteomyelitis.
Osteomyelitis is a common late-stage sequela of deep heel ulceration and frequently results in leg amputation, because few options exist to effectively treat this challenging problem. A technique that has been successful at our institution is near total calcanectomy with rotational flap closure of large decubitus heel ulcers complicated by calcaneal osteomyelitis. A case series is presented of 3 patients with differing locations or "zones" of heel decubitus ulceration with acute osteomyelitis. A description of our preferred flap choice, given the zone of the ulceration, is demonstrated, as are the surgical principles for the treatment approach. This procedure is an effective option, provided specific principles are followed to achieve complete wound coverage and minimize pressure points both when in bed and when walking.


Agreement between physicians' and patients' ratings on the Montgomery-Åsberg Depression Rating Scale.
Self-rating scales developed for monitoring depression severity are potentially informative and cost effective tools. There is an increasing tendency to use the Montgomery-Åsberg Depression Rating Scale (MADRS) and the self-rating version (MADRS-S) interchangeably. 400 patients with major depressive disorder were included. Concordance between patient and physician ratings was measured by means of repeated MADRS and MADRS-S ratings during a six-month drug trial and one-year follow-up. Overall scores from patients and physicians show the same trends and both are sensitive to improvements. Our results, however, show only moderate to good agreement between patient and physician ratings. Intraclass coefficients ranged from 0.47 to 0.75 with highest agreement at week 8. Generalizability is restricted to outpatients in general practice with moderate to severe depression. MADRS-S and MADRS scale definitions are similar but not identical concerning language and are scaled differently, 0-6 vs. 0-3, respectively, which may have influenced the results. The exclusion criteria restricted the range of values for the item Suicidal thoughts/Zest for life, which may have reduced the correlations. MADRS-S is a suitable tool for following patients' symptoms on a regular basis over time and may also be used to compensate for bias in physicians' ratings in drug trials.


Isopropanol exposure: environmental and biological monitoring in a printing works.
Occupational exposure to isopropanol was studied in 12 workers by testing environmental air, alveolar air, venous blood, and urine during their work shift. Isopropanol, which ranged in environmental air between 7 and 645 mg/m3, was detected in alveolar air, where it ranged between 4 and 437 mg/m3, but not in blood or in urine. Alveolar isopropanol concentration (Ca) was significantly correlated with environmental isopropanol concentration (Ci) at any time of exposure. The value of the arithmetical Ca/ci ratio was 0.418 (SD 0.101). Acetone, which is a metabolite of isopropanol, was found in alveolar air, blood, and urine in concentrations that were higher during exposure than before. Alveolar and blood acetone concentrations were highly correlated with alveolar isopropanol concentrations at any time during exposure. Acetone ranged between 0.76 and 15.6 mg/l in blood, between 4 and 93 micrograms/l in alveolar air, and between 0.85 and 53.7 mg/l in urine. Alveolar (Ca) and blood (Cb) acetone concentrations were highly correlated (r = 0.67), with a Cb/Ca ratio of 101. Alveolar isopropanol uptake ranged between 0.03 and 6.8 mg/min and was highly correlated with environmental isopropanol concentration (r = 0.92). During exposure, acetone eliminated by the lungs ranged between 20 and 273 mg in seven hours and in urine between 0.3 and 9.6 mg in seven hours. Acetonuria was higher the next morning than at the end of exposure.


Diagnosing Depression in Chronic Pain Patients: DSM-IV Major Depressive Disorder vs. Beck Depression Inventory (BDI).
Diagnosing depression in chronic pain is challenging due to overlapping somatic symptoms. In questionnaires, such as the Beck Depression Inventory (BDI), responses may be influenced more by pain than by the severity of depression. In addition, previous studies have suggested that symptoms of negative self-image, a key element in depression, are uncommon in chronic pain-related depression. The object of this study is to assess the relationship of the somatic and cognitive-emotional items of BDI with the diagnosis of depression, pain intensity, and disability. One hundred consecutive chronic pain patients completed the Structured Clinical Interview for DSM Disorders (SCID) for the diagnosis of major depressive disorder (MDD) according to DSM-IV. Two subscales of BDI (negative view of self and somatic-physical function) were created according to the factor model presented by Morley. In the regression analysis, the somatic-physical function factor associated with MDD, while the negative view of self factor did not. Patients with MDD had higher scores in several of the BDI items when analysed separately. Insomnia and weight loss were not dependent on the depression diagnosis. The relatively small sample size and the selected patient sample limit the generalisability of the results. Somatic symptoms of depression are also common in chronic pain and should not be excluded when diagnosing depression in pain patients. Regardless of the assessment method, diagnosing depression in chronic pain remains a challenge and requires careful interpretation of symptoms.


Shine on: achieving career satisfaction as a registered nurse.
This phenomenological study focuses on the experience of career satisfaction among registered nurses. Potential participants were asked, "Do you love your work as a nurse?" A random sample of eight nurses who answered yes to this question was questioned further during semistructured conversations. Conversations were recorded and transcribed. Data collected were in the form of descriptions of times during the participants' careers when they felt most professionally fulfilled. Through narrative and poetic analysis, themes of "upholding the vulnerable," "going the extra mile," and "attending to the essential ordinary" were identified. Nurse educators play an important role facilitating career satisfaction for registered nurses. Practical implications for continuing education for educators and practicing nurses are addressed.