[Osteoporosis: multidisciplinary and radiologic aspects].
Osteoporosis in one of the most common diseases among the elderly individuals. Bone hardness is reduced to the critically low level. Even the smallest trauma makes previously healthy individual severely ill patient. In the USA, 1.6 million people sustain fractures due to osteoporosis each year, while approximately 14 billion dollars is spent on their treatment. The costs of osteoporosis treatment exceed the costs of prevention. The studies on the incidence, prevalence and socioeconomic aspects of osteoporosis, its prevention and treatment are expected to be carried out in our country in the future. Prevention of osteoporosis is not an individual problem, but it has wide social implications. Osteoporosis may be prevented and further loss of bone substance may be discontinued.


Binding and functional effects of atrial natriuretic factor in isolated rat kidney.
A new methodological approach was developed to study the relationship between specific binding and dose-response curves of the renal effects of atrial natriuretic factor (ANF) in isolated perfused rat kidneys (IK). IK were perfused with 125I-labeled and unlabeled ANF 1-28 (4 pM to 1 microM) to determine the following: 1) distribution, capacity (Cmax), and apparent affinity (S50) of specific binding of ANF 1-28 in cortex, outer medulla, and papilla and 2) dose-response curves of the effects of ANF 1-28 on renal hemodynamics and excretion of fluid and electrolytes. The kidney had a very high density of high-affinity binding sites for ANF. Cortex had greater than 90% of total binding sites (Cmax = 6.8 pmol/g tissue; S50 = 54 pM), whereas papilla had less than 2% of total binding sites with a 10-fold lower apparent affinity (S50 = 860 pM) than in cortex. ANF-induced increases in glomerular filtration rate and excretion of fluid and electrolytes were detectable at 10-100 pM and maximal effects occurred at 1-10 nM ANF. Below 1 nM there was no dissociation between the renal hemodynamic and natriuretic effects of ANF. There was a close agreement between dose-response and binding curves of ANF to cortex. Results demonstrate that binding site occupancy in kidney cortex and renal effects of ANF occur at near physiological concentrations of the hormone.


Convergent effects of mouse Pet-1 deletion and human PET-1 variation on amygdala fear and threat processing.
Serotonin is critical for shaping the development of neural circuits regulating emotion. Pet-1 (FEV-1) is an ETS-domain transcription factor essential for differentiation and forebrain targeting of serotonin neurons. Constitutive Pet-1 knockout (KO) causes major loss of serotonin neurons and forebrain serotonin availability, and behavioral abnormalities. We phenotyped Pet-1 KO mice for fear conditioning and extinction, and on a battery of assays for anxiety- and depression-related behaviors. Morphology of Golgi-stained neurons in basolateral amygdala (BLA) and prelimbic cortex was examined. Using human imaging genetics, a common variant (rs860573) in the PET-1 (FEV) gene was tested for effects on threat-related amygdala reactivity and psychopathology in 88 Asian-ancestry subjects. Pet-1 KO mice exhibited increased acquisition and expression of fear, and elevated fear recovery following extinction, relative to wild-type (WT). BLA dendrites of Pet-1 KO mice were significantly longer than in WT. Human PET-1 variation associated with differences in amygdala threat processing and psychopathology. This novel evidence for the role of Pet-1 in fear processing and dendritic organization of amygdala neurons and in human amygdala threat processing extends a growing literature demonstrating the influence of genetic variation in the serotonin system on emotional regulation via effects on structure and function of underlying corticolimbic circuitry.


Combined chemoradiotherapy with gemcitabine in patients with locally advanced inoperable transitional cell carcinoma of the urinary bladder and/or in patients ineligible for surgery: a phase I trial.
We conducted a phase I trial of gemcitabine (gem) with concurrent radiotherapy in patients with muscle-invasive bladder cancer (BC) ineligible for surgery or cisplatin or refusing organ loss. Patients with urothelial cancer, cT2-T4, cN0-1, M0, ineligible for surgery due to local tumor extension, PS, age or co-morbidities or who refused surgery were included. After maximal transurethral resection, the treatment schedule included: twice-weekly i.v. infusion of gem [dose levels (DL) 1-6: 20, 27, 30, 33, 50 and 40 mg/m(2), respectively] for 30 min and concurrent radiotherapy (RT) to the bladder with 55.5 Gy. The primary end point was to determine the maximum-tolerated dose (MTD) and the dose recommended (RD) for further studies of this gem schedule. The secondary end point was late toxicity. The MTD was defined by dose-limiting toxicity (DLT) in 2 or more of 6 patients, discontinuation of RT and/or gem for >1 week in 2 or more of 6 patients due to grade (G) 3/4 acute and/or late toxicity in more than 2 of 18 patients. Thirty-five of 44 patients were assessable for toxicity and thus the primary end point. DLTs occurred in two of five patients at dose level 5: one G3 alanine aminotransferase elevation and one G3 fatigue. The MTD, therefore, was 50 mg/m(2) gem twice weekly. At DL 6 with 40 mg/m(2), the RD was established: only one of six patients developed G3 fatigue and diarrhea. Late toxicity was rare and of low grade (only G1-2). The 2-year locoregional failure rate was 32% (9/28); 10 of 28 patients (38%) were alive with an intact bladder and no evidence of recurrent disease, 9 patients developed distant metastases and 6 died of their disease. Gemcitabine in combination with RT is well tolerated in BC patients ineligible for surgery and/or cisplatin. The RD of gemcitabine for subsequent trials is 40 mg/m(2) twice weekly with concurrent radiation.


Effect of acute doses of 2-acetylaminofluorene on the capacity of rat liver to repair methylated purines in DNA in vivo and in vitro.
Male Wistar rats were given various doses of 2-acetylaminofluorene (AAF) at doses of 0.74, 2.22, 6.67, or 20 mg/kg i.p. 24 hr before administration of [14C]dimethylnitrosamine (1 or 2 mg/kg i.p.). Analysis of liver DNA isolated from animals killed 5 hr later showed variations between groups treated with different amounts of AAF in the amounts of 3-methyladenine, 7-methylguanine, and O6-methylguanine (O6-mGua). However, the relative amounts of these products were unchanged by AAF pretreatment except after 20 mg/kg when the reduced O6-mGua:7-methylguanine ratio indicated enhanced O6-mGua repair. Specific enhancement of O6-mGua repair was also found 5 hr after administration of [14C]methylnitrosourea (11.5 mg/kg) to animals pretreated with AAF (20 mg/kg), while the amounts of O6-mGua in liver ribosomal RNA afer [14C]dimethylnitrosamine were unaffected by this AAF dose. Pretreatment of rats with AAF 29 hr earlier increased the capacity of cell-free liver extracts to remove O6-mGua from [3H]methylnitrosourea-methylated DNA in vitro. The increase was detectable after 2.22 mg/kg and reached a maximum 3.5-fold increase after AAF, (60 mg/kg). 7-Methylguanine and 3-methyladenine-DNA glycosylase activities were also increased, but this was independent of the dose of AAF. AAF pretreatment produced a slight (3- to 4-fold) increase in incorporation of [3H]thymidine or labeled one-carbon breakdown products of [14C]dimethylnitrosamine into liver DNA which appeared to parallel in vitro O6-mGua repair enhancement, but the increased [3H]thymidine uptake was statistically significant only after the 60-mg/kg dose.


Comparison of Urine and Plasma Biomarker Concentrations Measured by Aptamer-Based versus Immunoassay Methods in Cardiac Surgery Patients.
Protein detection assays are invaluable tools in the field of biomarker discovery. However, only immunoassays are widely used and can measure 10-20 analytes per biosample. The novel SOMAmer-based assay uses nucleotide aptamer technology to measure over 1300 analytes per biosample. We compared the SOMAmer-based platform to traditional approaches to quantify analytes in a clinical setting with paired samples before and after cardiac surgery. In a substudy of the Translational Research Investigating Biomarker Endpoints in Acute Kidney Injury cohort, 54 individuals with acute kidney injury after cardiac surgery were identified. Preoperative and postoperative plasma and urine samples that had been previously evaluated for biomarker concentrations via immunoassays were analyzed via SOMAmer-based assay. Spearman correlations were estimated when >50% of biomarker values were within detectable ranges by immunoassay (plasma biomarkers: preoperative, 26/33; postoperative, 31/33; urine biomarkers: preoperative, 13/16; postoperative, 16/16). Overall, 27% of reportable plasma preoperative biomarkers displayed correlations ≥0.75 between immunoassay and SOMAmer measurements; 23% displayed correlations of 0.50-0.75, and 50% displayed correlations <0.50. In urine these values were 15%, 39%, and 46%, respectively. Forty-two percent of reportable plasma postoperative biomarkers displayed correlations ≥0.75, 16% displayed correlations 0.50-0.75, and 42% displayed correlations <0.50. In urine, these values were 19%, 25%, and 56%, respectively. In cardiac surgery patients, the SOMAmer-based assay detects proteins with moderate to strong correlation to current immunoassay methods. The correlations in urine are weaker than those in plasma. SOMAmer-based assay technology should be further evaluated in multiple settings as a high-throughput screening tool for biomarker discovery.


Reduced susceptibility to ventricular tachyarrhythmias in rats selectively bred for high aerobic capacity.
Reperfusion after a brief period of cardiac ischemia can lead to potentially lethal arrhythmias. Human epidemiological studies and experimental work with animals indicate that regular physical activity is associated with reductions in cardiovascular disease (CVD) risk factors and sudden cardiac death. Similarly, artificial selection of rats for high aerobic treadmill-running capacity (high-capacity runners; HCR) has been shown to reduce CVD risk factors relative to rats selected as low-capacity runners (LCR). Therefore, we tested the hypothesis that HCR, relative to LCR rats, would be less susceptible to ischemia-reperfusion-mediated ventricular tachyarrhythmias. To test this hypothesis, we measured the susceptibility to ventricular tachyarrhythmias produced by 3 min of occlusion and reperfusion of the left main coronary artery in conscious LCR and HCR rats. Results document a significantly lower incidence of ventricular tachyarrhythmias in HCR (3 of 11, 27.3%) relative to LCR (6 of 7, 85.6%) rats. The decreased susceptibility to tachyarrhythmias in HCR rats was associated with a reduced cardiac metabolic demand during ischemia (lower rate-pressure product and ST segment elevation) as well as a wider range for the autonomic control of heart rate. The HCR and LCR represent a unique substrate for evaluation of the mechanisms underlying ischemia-mediated cardiac arrhythmogenesis.


Deletions involving long-range conserved nongenic sequences upstream and downstream of FOXL2 as a novel disease-causing mechanism in blepharophimosis syndrome.
The expression of a gene requires not only a normal coding sequence but also intact regulatory regions, which can be located at large distances from the target genes, as demonstrated for an increasing number of developmental genes. In previous mutation studies of the role of FOXL2 in blepharophimosis syndrome (BPES), we identified intragenic mutations in 70% of our patients. Three translocation breakpoints upstream of FOXL2 in patients with BPES suggested a position effect. Here, we identified novel microdeletions outside of FOXL2 in cases of sporadic and familial BPES. Specifically, four rearrangements, with an overlap of 126 kb, are located 230 kb upstream of FOXL2, telomeric to the reported translocation breakpoints. Moreover, the shortest region of deletion overlap (SRO) contains several conserved nongenic sequences (CNGs) harboring putative transcription-factor binding sites and representing potential long-range cis-regulatory elements. Interestingly, the human region orthologous to the 12-kb sequence deleted in the polled intersex syndrome in goat, which is an animal model for BPES, is contained in this SRO, providing evidence of human-goat conservation of FOXL2 expression and of the mutational mechanism. Surprisingly, in a fifth family with BPES, one rearrangement was found downstream of FOXL2. In addition, we report nine novel rearrangements encompassing FOXL2 that range from partial gene deletions to submicroscopic deletions. Overall, genomic rearrangements encompassing or outside of FOXL2 account for 16% of all molecular defects found in our families with BPES. In summary, this is the first report of extragenic deletions in BPES, providing further evidence of potential long-range cis-regulatory elements regulating FOXL2 expression. It contributes to the enlarging group of developmental diseases caused by defective distant regulation of gene expression. Finally, we demonstrate that CNGs are candidate regions for genomic rearrangements in developmental genes.


How co-locating public mental health interventions in community settings impacts mental health and health inequalities: a multi-site realist evaluation.
Public mental health interventions are non-clinical services that aim to promote wellbeing and prevent mental ill health at the population level. In England, the health, social and community system is characterised by complex and fragmented inter-sectoral relationships. To overcome this, there has been an expansion in co-locating public mental health services within clinical settings, the focus of prior research. This study evaluates how co-location in community-based settings can support adult mental health and reduce health inequalities. A qualitative multi-site case study design using a realist evaluation approach was employed. Data collection took place in three phases: theory gleaning, parallel testing and refining of theories, and theory consolidation. We collected data from service users (n = 32), service providers (n = 32), funders, commissioners, and policy makers (n = 11), and members of the public (n = 10). We conducted in-depth interviews (n = 65) and four focus group discussions (n = 20) at six case study sites across England, UK, and two online multi-stakeholder workshops (n = 20). Interview guides followed realist-informed open-ended questions, adapted for each phase. The realist analysis used an iterative, inductive, and deductive data analysis approach to identify the underlying mechanisms for how community co-location affects public mental health outcomes, who this works best for, and understand the contexts in which co-location operates. Five overarching co-location theories were elicited and supported. Co-located services: (1) improved provision of holistic and person-centred support; (2) reduced stigma by creating non-judgemental environments that were not associated with clinical or mental health services; (3) delivered services in psychologically safe environments by creating a culture of empathy, friendliness and trust where people felt they were being treated with dignity and respect; (4) helped to overcome barriers to accessibility by making service access less costly and more time efficient, and (5) enhance the sustainability of services through better pooling of resources. Co-locating public mental health services within communities impacts multiple social determinants of poor mental health. It has a role in reducing mental health inequalities by helping those least likely to access services. Operating practices that engender inter-service trust and resource-sharing are likely to support sustainability.


[(35)S]TBPS binding to purified benzodiazepine-GABA-ionophore receptor complex: the effect of GABA on the modulation exerted by benzodiazepine site ligands.
Benzodiazepine GABA-ionophore receptor complex ligands showed persistent modulation of the chloride ionophore, labeled by [(35)S]TBPS, even after receptor complex extensive purification. GABA caused inhibition of [(35)S]TBPS binding, while benzodiazepine agonists increased and benzodiazepine inverse agonists decreased the specific [(35)S]TBPS binding to the purified receptor. When GABA binding sites were occupied by the neurotransmitter benzodiazepine receptor agonists and antagonists reversed their effects clonazepam in fact inhibited and ?-carboline ethyl ester increased [(35)S]TBPS binding in the presence of GABA. The antagonist flumazenil showed no effect both in the presence or absence of GABA.


Alcohol, But Not Depression or IPV, Reduces HIV Adherence Among South African Mothers Living with HIV Over 5 Years.
Alcohol, depression, and intimate partner violence (IPV) are endemic in sub-Saharan Africa. This article examines whether and how these conditions affect mothers living with HIV (MLH), compared to mothers without HIV (MWOH). In particular, we assess the influence of these comorbidities on engagement in HIV care and adherence to antiretroviral therapies (ARV) among MLH. Data on maternal HIV care are typically based on clinic samples, with substantial loss to follow-up. This study fills that gap by including all mothers in specified areas. A cohort study examines MLH in Cape Town, South Africa recruited in pregnancy and followed repeatedly for 5 years, compared to MWOH. Almost all (98%) pregnant women in 12 neighborhoods (N = 594) were recruited in pregnancy. Mothers and children were reassessed five times over 5 years with high retention rates at each of the six assessments, from 98.7% at 2 weeks to 82.8% at 5 years post-birth. MLH's uptake and adherence to HIV care was evaluated over time associated with maternal comorbidities of alcohol use, depressed mood, and IPV using mixed effects logistic regression. MLH have fewer resources (income, food, education) and are more likely to face challenges from alcohol, depression, and having seropositive partners over time than MWOH. Only 22.6% of MLH were consistently engaged in HIV care from 6 months to 5 years post-birth. At 5 years, 86.7% self-reported engaged in HIV care, 76.9% were receiving ARVs and 87% of those on ARV reported consistent ARV adherence. However, data on viral suppression are unavailable. Alcohol use, but not depressed mood or IPV, was significantly related to reduced uptake of HIV care and adherence to ARV over time. Adherence to lifelong ARV by MLH requires a combination of structural and behaviorally-focused interventions. Alcohol abuse is not typically addressed in low and middle-income countries, but is critical to support MLH.


Sexual Assault Among Women in College: Immediate and Long-Term Associations With Mental Health, Psychosocial Functioning, and Romantic Relationships.
The current study sought to examine immediate and long-term consequences of college sexual assault (C-SA) among women with no prior sexual assault history. While much is known regarding the short-term negative impact of C-SA, the current study examines whether C-SA is associated with immediate academic and psychosocial consequences as well as long-term poorer mental health (depression, posttraumatic stress [PTS], anxiety) and interpersonal functioning (relationship quality, sexual and emotional intimacy). In addition, the current study explores potential moderators of these associations, including race, the nature of the assault, resulting injury, relation to perpetrator, and whether the assault was reported. A stratified design was used comparing women who experienced C-SA (n = 201) to women with no C-SA history (n = 203) controlling for age, education, race, and ethnicity. Results from a series of repeated-measures analyses of variance (ANOVAs) demonstrated that across race and ethnicity, women with a history of C-SA reported lower grade-point averages, more missed classes, and fewer serious romantic relationships in college following the assault. Furthermore, results from a series of linear and logistic regression revealed that approximately 9 years later, women who experienced C-SA reported greater symptoms of depression, anxiety, and PTS as well as lower emotional and sexual intimacy. These associations differed by a number of assault variables (assault type, relation to perpetrator, amount of fear reported, physical injuries sustained, whether the assault was reported, whether medical treatment was sought). The current study further confirms the significant and pervasive impact of C-SA associated with women's health and functioning, warranting further intervention to both reduce the incidence of C-SA and expand the reach of existing mental health interventions to survivors.


[Artificial embolization of cerebral arteriovenous aneurysms via the vertebro-basilar system].
Data are presented on artificial embolization of arterio-venous aneurysms of the brain via the vertebro-basilar system in 12 patients. In one case the embolization was performed by way of vertebral artery catheterization through the axillary artery, in the remaining II cases--through the femoral artery. The embolization was carried out with the aid of polystyrene balls, 2 to 3 mm in diameter, numbering 8 to 40. In some cases the needed direction of the emboli movement was achieved by applying a magnetic field. No complications were encountered in the embolization via the vertebral system.


Topical application of mitomycin C prevents epineural scar formation in rats.
The role of topically applied mitomycin C in preventing postoperative perineural fibrosis was examined by gross anatomical dissection and histological analysis in rats. The sciatic nerve was exposed bilaterally in 24 Wistar adult male rats, and an abrasion injury was produced on the exposed surface of the biceps femoris muscle in all animals. In the experimental group, cotton pads soaked with mitomycin C (0.5 mg/ml) were placed around the nerves for 5 min, whereas cotton pads soaked with saline were applied to the control group. Four weeks after surgery, the neurolysis sites were evaluated by blinded surgical dissection. Perineural adhesions were graded using a numerical grading scheme. The scar tissue formation index was also calculated, and a grading was made according to the number of fibroblasts/fibrocytes counted around the epineurium in histological evaluation. Mitomycin C-treated nerves showed significantly less perineural adhesions than controls. Quantification of the dense connective tissue surrounding the nerves revealed a statistically significant reduction around nerves treated with mitomycin C, and the number of fibroblast/fibrocytes was also significantly reduced. Application of topical mitomycin C might be effective in preventing epineural scar formation after neurolysis of peripheral nerves.


[A method of objective evaluation of pain].
Methods of objective evaluation of pains suggested before require the use of different outfits. In some cases this circumstance makes it difficult to employ them in diagnostic practice. In view of this the author suggests a new method deprived of the above-indicated shortcoming. It is reduced to the application of drops of iodine solution in dimexide to the painful area and outside it. Asynchronous changes in the intensity of the staining of the spots formed as a result of drop smearing on the skin surface in the painful area and outside it confirm the presence of pains.


Role of calcium in the interaction of alpha and beta adrenoceptor-mediated renin release in isolated, constant pressure perfused rabbit kidneys.
These experiments were designed to study the role of calcium in the modulation of renin secretion by alpha and beta adrenoceptors. Rabbit kidneys were isolated and single-pass perfused with a modified Ringer's solution. Renal perfusion pressure was precisely controlled by an electronic servocontrol system. Tubular events were minimized by ligation of the ureter before initiating the studies. Under these conditions the predominant factor modifying renin secretion was assumed to originate directly on the juxtaglomerular cells. Isoproterenol infused at 0.1, 0.5, 1.0 and 5.0 nM/min/g of kidney weight increased renin secretion in a dose-dependent manner whereas phenylephrine infused at identical molar doses did not. In addition, phenylephrine (5.0 nM/min/g of kidney weight) blocked the usual response to isoproterenol. Removal of calcium from the perfusing medium had no effect on either the response to isoproterenol or the lack of a response to phenylephrine. On the other hand, when calcium is removed from the perfusate or when D-600, a calcium channel blocker, is added to calcium-containing medium, phenylephrine failed to block the usual response to isoproterenol. We conclude that the suppression of beta adrenoceptor stimulation of renin release by alpha adrenoceptor agonists is calcium dependent by a final mechanism as yet undefined, but probably involving movement of calcium into the juxtaglomerular cells.


Cell migration in the Xenopus gastrula.
Xenopus gastrulation movements are in large part based on the rearrangement of cells by differential cell-on-cell migration within multilayered tissues. Different patterns of migration-based cell intercalation drive endoderm and mesoderm internalization and their positioning along their prospective body axes. C-cadherin, fibronectin, integrins, and focal contact components are expressed in all gastrula cells and play putative roles in cell-on-cell migration, but their actual functions in this respect are not yet understood. The gastrula can be subdivided into two motility domains, and in the vegetal, migratory domain, two modes of cell migration are discerned. Vegetal endoderm cells show ingression-type migration, a variant of amoeboid migration characterized by the lack of locomotory protrusions and by macropinocytosis as a mechanism of trailing edge resorption. Mesendoderm and prechordal mesoderm cells use lamellipodia in a mesenchymal mode of migration. Gastrula cell motility can be dissected into traits, such as cell polarity, adhesion, mobility, or protrusive activity, which are controlled separately yet in complex, combinatorial ways. Cells can instantaneously switch between different combinations of traits, showing plasticity as they respond to substratum properties. This article is categorized under: Early Embryonic Development > Gastrulation and Neurulation.


Hierarchically structured bioactive foams based on polyvinyl alcohol-sepiolite nanocomposites.
Hierarchically structured polymer-clay foams comprised of sepiolite and polyvinyl alcohol (PVA) are functionalized with enzymatic and conductive properties to be employed as bioreactors and 3D bioelectrodes. The concept of nanoarchitectonics was followed in the design of this hierarchical and multifunctional cellular material through tailoring of the properties at different length scales, interfacial properties of the sepiolite fibres through adsorption of lipid molecules, bioactivity by supramolecular assembly of urease enzymes on the sepiolite-lipid bio-nanohybrid, and meso- and macroporosity controlled by incorporation of the clay fibres into a PVA matrix, and further cross-linking with borax and ice-templating. The resultant bionanocomposite foams show high structural integrity, tailored multi-level porosity, and biocompatibility which is manifested in sustained enzymatic activity. Exploratory studies show that additional doping with carbon black rendered conductive foams that could hence find usage in bioelectrocatalysis.


Regional chromosome mapping of human collagen genes alpha 2(I) and alpha 1(I) (COLIA2 and COLIA1).
For the assignment of the genes for the pro-alpha 2(I) (COLIA2) and the pro-alpha 1(I) (COLIA1) collagens, cDNA and genomic DNA probes were used in in situ hybridization experiments on human prometaphase chromosomes. An improved staining method is reported for the simultaneous identification of chromosomes and the autoradiographic grains after the hybridization procedures. With this procedure more cells with higher resolution could be used for the assignment of genes by in situ hybridization. Statistical analysis of the grains located on respectively 660 and 302 metaphases using pro-alpha 2(I) and pro alpha 1(I) DNA probes, confirmed the assignment of these genes to human chromosomes 7 and 17. Analysis of the grain distribution on prometaphase chromosomes showed that the location of the pro-alpha 2(I) collagen gene is in the region 7q21.3-22.1. The location of the pro-alpha 1(I) collagen gene was found to be in band 17q21.31-2205.


[Prevalence and factors associated with conducting screening tests for prostate cancer in the elderly in Juiz de Fora in the state of Minas Gerais, Brazil].
Little is known about screening practices for prostate cancer in elderly Brazilians. The aim of this study was to estimate the prevalence of screening tests for prostate cancer in the elderly in Juiz de Fora in the state of Minas Gerais and to analyze the associated factors. This is a cross-sectional study with 2825 men aged 60 years or older who participated in the vaccination against influenza campaign in 2006. Social and demographic variables and others related to health conditions and the use of health services were analyzed. Poisson regression multivariate analysis was used to analyze associations between the covariates and the dependent variables and the prevalence of the tests was estimated. The median age was 70.0 (± 7.2) years. The prevalence of performing digital rectal examination was 61.0% and 75.5% for PSA. The "family history of prostate cancer," "type of health service," "marital status," "regular use of medication" and "education" variables were independent factors associated with performing digital rectal examination. The same variables, with the exception of "marital status" remained in the multiple model for PSA. The study showed that many seniors have adopted the practice of screening and the need to measure and describe this process in view of its possible repercussions on public health.


HIV-positive in the darkness of a correctional facility: more vulnerable and less treated.
In French Guiana, 4.5-5.0% of HIV-infected persons experienced an incarceration between 2007 and 2013. A cross-sectional survey was performed to describe the epidemiology and treatment outcomes of a population of HIV-infected inmates in French Guiana. The study population was patients released between 1/2007 and 12/2013, after >30 days of incarceration (n = 147). A secondary objective aimed to identify its main specificities, relative to both the general inmate population and the nonincarcerated HIV population. The socioeconomic situation of HIV-infected inmates was particularly precarious, relative to other detainees: 58.1% had never attended school (versus 5.5%, p < 0.01), 31.0% were homeless (versus 8.5%, p < 0.01), 63.9% were repeat offenders (versus 46.6%, p < 0.01), 33.3% were crack cocaine users (versus 9.8-12%, p < 0.01). The frequency of hypertension and chronic B hepatitis was also higher. Only 50.3% of inmates were on antiretroviral therapy (ART) versus 92.6% in the hospital HIV population (p < 0.001). Among untreated patients, 15.1% refused ART. Among those treated, 81.3% were virologically suppressed. Although comparisons were biased, HIV-positive inmates had more psychosocial vulnerabilities than the general inmate population. Despite ART availability and excellent treatment outcomes, undertreatment was a fact not completely explained by patient refusal. HIV-infected inmates should benefit from increased attention by health care and social workers.


Renal cell carcinoma, occupational pesticide exposure and modification by glutathione S-transferase polymorphisms.
This study investigated associations between occupational pesticide exposure and renal cell carcinoma (RCC) risk. To follow-up on a previous report by Buzio et al., we also considered whether this association could be modified by glutathione S-transferase M1 and T1 (GSTM1 and GSTT1) genotypes. About 1097 RCC cases and 1476 controls from Central and Eastern Europe were interviewed to collect data on lifetime occupational histories. Occupational information for jobs held for at least 12 months duration was coded for pesticide exposures and assessed for frequency and intensity of exposure. GSTM1 and GSTT1 gene deletions were analyzed using TaqMan assays. A significant increase in RCC risk was observed among subjects ever exposed to pesticides [odds ratio (OR): 1.60; 95% confidence interval (CI): 1.00-2.55]. After stratification by genotypes, increased risk was observed among exposed subjects with at least one GSTM1 active allele (OR: 4.00; 95% CI: 1.55-10.33) but not among exposed subjects with two GSTM1 inactive alleles compared with unexposed subjects with two inactive alleles (P-interaction: 0.04). Risk was highest among exposed subjects with both GSTM1 and GSTT1 active genotypes (OR: 6.47; 95% CI: 1.82-23.00; P-interaction: 0.02) compared with unexposed subjects with at least one GSTM1 or T1 inactive genotype. In the largest RCC case-control study with genotype information conducted to date, we observed that risk associated with pesticide exposure was exclusive to individuals with active GSTM1/T1 genotypes. These findings further support the hypothesis that glutathione S-transferase polymorphisms can modify RCC risk associated with occupational pesticide exposure.


The Prototypes of Tobacco Users Scale (POTUS) for Cigarette Smoking and E-Cigarette Use: Development and Validation.
Endorsing prototypes of cigarette smokers predicts cigarette smoking, but less is known about prototypes of users of other tobacco products. Our study sought to establish the reliability and validity of a measure of prototypes of smokers and e-cigarette users. Participants were from a national survey of smokers and non-smokers (n = 1414), a randomized clinical trial (RCT) of adult smokers (n = 2149), and adolescent children of adults in the trial (n = 112). The Prototypes of Tobacco Users Scale (POTUS) has four positive adjectives (cool, sexy, smart, and healthy) and four negative adjectives (disgusting, unattractive, immature, and inconsiderate) describing cigarette smokers and e-cigarette users. Confirmatory factor analyses identified a two-factor solution. The POTUS demonstrated strong internal consistency reliability in all three samples (median α = 0.85) and good test-retest reliability among adults in the RCT (median r = 0.61, 1-4 weeks follow-up). In the RCT, smokers more often agreed with negative prototypes for smokers than for e-cigarette users (mean = 2.03 vs. 1.67, p < 0.05); negative prototypes at baseline were also associated with more forgoing of cigarettes and making a quit attempt at the end of the trial (Week 4 follow-up). The POTUS may be useful to public health researchers seeking to design interventions that reduce tobacco initiation or cessation through the manipulation of tobacco user prototypes.


The role of miRNAs in cancer: from pathogenesis to therapeutic implications.
Cancer is still one of the dominating causes of deaths worldwide, although there have been important enhancements for detection and diagnosis of cancer recently. miRNAs are shown to participate in carcinogenesis of several types of tumors and their aberrant expression of miRNAs has been detected in cell lines, xenografts and clinical samples. miRNAs are thought to target and modulate the expression of more than 60% of human genes, which makes the expressional regulation by miRNAs the most abundant post-transcriptional regulation mode. Here, we have reviewed the most current literature to shed a light on the functions of miRNAs on human carcinogenesis. Possible roles of miRNAs in oncogenesis through both genetic and epigenetic changes occurring during cancer initiation, progression, invasion or metastasis are summarized.


Clear air turbulence: detection by infrared observations of water vapor.
"Forward-looking" infrared measurements of water vapor from the C-141A Kuiper Airborne Observatory of the National Aeronautics and Space Administration Ames Research Center show large, distinctly identifiable, signal anomalies from 4 to 10 minutes in advance of subsequent encounters with clear air turbulence (CAT). These anomalies are characteristically different from the signals not followed by CAT encounters. Results of airborne field trials in which the infrared radiometer was used indicate that, out of 51 situations, 80 percent were CAT alerts followed by CAT encounters, 12 percent were "false alarms" (CAT alerts not followed by CAT encounters), and 8 percent were CAT encounters not preceded by an infrared signal anomaly or CAT alert.


Ionic strength perturbation kinetics of gene 32 protein dissociation from its complex with single-stranded DNA.
Equilibrium and kinetic studies of the interaction of gene 32 protein of T4 phage with single-stranded fd DNA were performed monitoring the changes in protein fluorescence. From the fluorescence titrations, it was estimated that a monomer of gene 32 protein covered six nucleotide bases on the DNA and the lower limit for the apparent association constant was 1.9 x 10(8) M-1 with a cooperative parameter of 10(3) in 0.1 M 2-amino-2-hydroxymethyl-1,3-propanediol hydrochloride (pH 7) at 25 degrees C. When an ionic strength jump was applied to the gene 32 protein-fd DNA complex using a stopped-flow apparatus, the complex underwent a dissociation into its individual components accompanied by an increase in protein fluorescence. The kinetics of the dissociation are not consistent with a single first-order process. The data, however, can be analyzed in terms of a model in which gene 32 protein molecules release cooperatively starting from either one or both ends of a cluster of proteins bound to fd DNA. This type of dissociation of gene 32 protein from single-stranded DNA is very efficient and has interesting implications: it could provide a way to facilitate a rapid "zippering" of the two complementary DNA strands during DNA replication and genetic recombination.


Evolution of the microstructure and the drug release upon annealing the drug loaded lipid-surfactant microspheres.
The present study investigates the drug release-governing microstructural properties of melt spray congealed microspheres encapsulating the drug crystals in the matrix of glyceryl behenate and poloxamer (pore former). The solid-state, morphology, and micromeritics of the microspheres were characterized, before and after annealing, using calorimetry, X-ray scattering, porosimetry, scanning electron microscopy, and, NMR diffusometry. The in vitro drug release from and water uptake by the microspheres were obtained. The extent and the rate of drug release from the microspheres increased with a high poloxamer content and at higher annealing temperature and RH. All the drug release profiles were describable using the Higuchi release kinetics pointing towards the diffusion controlled release, both before and after annealing. The annealing process led to the polymorphic conversion of lipid and the increase in the pore size, predominantly at a higher temperature and humidity and for a high poloxamer content. The poloxamer domain increased from an initial 300 nm, up to 2000 nm upon annealing. The water diffusion rate inside the annealed microsphere was twice as fast as for unannealed counterparts. The findings relate the overall phase and pore structure change of the microsphere to the increased drug release induced by annealing. This work serves as a basis for the rational understanding of the modification of the in vitro performance by annealing, a widely used post-process for solid lipid products.


Studies on the bivalent-cation-activated ATPase activities of highly purified human platelet surface and intracellular membranes.
Membrane-bound Ca2+-ATPases are responsible for the energy-dependent transport of Ca2+ across membrane barriers against concentration gradients. Such enzymes have been identified in sarcoplasmic reticulum of muscle tissues and in non-muscle cells in both surface membranes and endoplasmic-reticulum-like intracellular membrane complexes. In a previous study using membrane fractionation by density-gradient and free-flow electrophoresis, we reported that the intracellular membranes of human blood platelets were a major storage site for Ca2+ and involved in maintaining low cytosol [Ca2+] in the unactivated cell. In the present report we demonstrated that the intracellular membranes also exhibit a high-affinity Ca2+-ATPase which appears to be kinetically associated with the Ca2+-sequestering process. We found that both the surface membrane and the intracellular membrane exhibited a basal Mg2+-ATPase activity, but Ca2+ activation of this enzyme was confined only to the intracellular membrane. Use of Ca2+-EGTA buffers to control the extravesicle [Ca2+] allowed a direct comparison of the Ca2+-ATPase and the Ca2+-uptake process over a Ca2+ range of 0.01 microM to 1.0 mM, and it was found that both properties were maximally expressed in the range of external [Ca2+] 1-50 microM, with concentrations greater than 100 microM showing substantial inhibition. Double-reciprocal plots for the Ca2+-ATPase activity and Ca2+ uptake gave apparent Km values for Ca2+ of 0.15 and 0.13 microM respectively. However, similar plots for ATP with the enzyme revealed a discontinuity (two affinity sites, with Km 20 and 145 microM), whereas plots for the Ca2+ uptake gave a single Km value for Ca2+, 1.1 microM. Phosphorylation studies during Ca2+ uptake using [gamma-32P]ATP revealed two components of 90 and 95 kDa phosphorylated at extravesicle [Ca2+] of 3 microM. The Ca2+-ATPase activity, Ca2+ uptake and phosphorylation were all almost completely inhibited in the presence of 500 microM-Ca2+. Similar studies using mixed membranes revealed four other phosphoproteins (50, 40, 20 and 18 kDa) formed in addition to the 90 and 95 kDa components. The findings are discussed in the context of platelet Ca2+ mobilization for function and the mechanisms whereby Ca2+ homoeostasis is controlled in the unactivated cell.


Practices and risks associated with operation of tie-down lashings in the vehicle transport industry.
Load restraint systems in automobile transport utilise tie-down lashings placed over the car's tyres, which are tensioned manually by the operator using a ratchet assembly. This process has been identified as a significant manual handling injury risk. The aim of this study was to gain insight on the current practices associated with tie-down lashings operation, and identify the gaps between current and optimal practice. We approached this with qualitative and quantitative assessments and one numerical simulation to establish: (i) insight into the factors involved in ratcheting; (ii) the required tension to hold the car on the trailer; and (iii) the tension achieved by drivers in practice and associated joint loads. We identified that the method recommended to the drivers was not used in practice. Drivers instead tensioned the straps to the maximum of their capability, leading to over-tensioning and mechanical overload at the shoulder and elbow. We identified the postures and strategies that resulted in the lowest loads on the upper body during ratcheting (using both hands and performing the task with their full body). This research marks the first step towards the development of a training programme aiming at changing practice to reduce injury risks associated with the operation of tie-down lashings in the automobile transport industry. Practitioner Summary: The study investigated current practice associated with the operation of tie-down lashings through qualitative (interviews) and quantitative (biomechanical analysis) methods. Operators tended to systematically over-tension the lashings and consequently overexert, increasing injury risks.


A hierarchical Bayesian approach to multiple testing in disease mapping.
We propose a Bayesian approach to multiple testing in disease mapping. This study was motivated by a real example regarding the mortality rate for lung cancer, males, in the Tuscan region (Italy). The data are relative to the period 1995-1999 for 287 municipalities. We develop a tri-level hierarchical Bayesian model to estimate for each area the posterior classification probability that is the posterior probability that the municipality belongs to the set of non-divergent areas. We show also the connections of our model with the false discovery rate approach. Posterior classification probabilities are used to explore areas at divergent risk from the reference while controlling for multiple testing. We consider both the Poisson-Gamma and the Besag, York and Mollié model to account for extra Poisson variability in our Bayesian formulation. Posterior inference on classification probabilities is highly dependent on the choice of the prior. We perform a sensitivity analysis and suggest how to rely on subject-specific information to derive informative a priori distributions. Hierarchical Bayesian models provide a sensible way to model classification probabilities in the context of disease mapping.


Genetic and neural dissociation of individual responses to emotional expressions in human infants.
Interacting with others by interpreting and responding to their facial expressions is an essential and early developing social skill in humans. We examined whether and how variation in catechol-O-methyltransferase (COMT) and serotonin transporter (5-HTTLPR) genes is associated with 7-month-old infants' electrocortical responses to facial expressions. The results revealed that COMT variants are associated with differences in infants' brain responses to fearful faces over centro-parietal regions, whereas 5-HTTLPR variants are associated with differences in infants' brain responses to happy faces over fronto-temporal regions. Further support for differential associations of these gene variants with emotional processing came from our analysis of infant behavioral temperament: variation in COMT was associated with differences in infants' recovery from distress, whereas variation in 5-HTTLPR was associated with infants' smiling and laughter. This pattern of findings indicates that, in infancy, these genetic variants influence distinct brain systems involved in the processing of either positive or negative emotions. This has wide reaching implications for our understanding of how genetic variation biases specific brain mechanisms, giving rise to individual differences in emotional sensitivity and temperament.


UBE3C Facilitates the ER-Associated and Peripheral Degradation of Misfolded CFTR.
The ubiquitin E3 ligase UBE3C promotes the proteasomal degradation of cytosolic proteins and endoplasmic reticulum (ER) membrane proteins. UBE3C is proposed to function downstream of the RNF185/MBRL ER-associated degradation (ERAD) branch, contributing to the ERAD of select membrane proteins. Here, we report that UBE3C facilitates the ERAD of misfolded CFTR, even in the absence of both RNF185 and its functional ortholog RNF5 (RNF5/185). Unlike RNF5/185, UBE3C had a limited impact on the ubiquitination of misfolded CFTR. UBE3C knockdown (KD) resulted in an additional increase in the functional ∆F508-CFTR channels on the plasma membrane when combined with the RNF5/185 ablation, particularly in the presence of clinically used CFTR modulators. Interestingly, although UBE3C KD failed to attenuate the ERAD of insig-1, it reduced the ERAD of misfolded ∆Y490-ABCB1 and increased cell surface expression. UBE3C KD also stabilized the mature form of ∆F508-CFTR and increased the cell surface level of T70-CFTR, a class VI CFTR mutant. These results suggest that UBE3C plays a vital role in the ERAD of misfolded CFTR and ABCB1, even within the RNF5/185-independent ERAD pathway, and it may also be involved in maintaining the peripheral quality control of CFTR.


Production of interspecies chimeras by transplanting rosy barb (Puntius conchonius) embryonic cells to zebrafish (Danio rerio) embryos.
Establishment of a cell-mediated gene transfer system has potential as a new breeding technique for commercially valuable fishes. As an important step toward developing an inter-species chimera, cells from blastula-stage embryos of rosy barb (Puntius conchonius) were transplanted into zebrafish (Danio rerio) blastula-stage embryos to observe the development of the recipient. From the total of 473 transplants obtained only a fraction of 13 chimeras appeared perfectly normal after one month. Over two in normal 13 chimeras showed some characters from the donor cells with scarce pigmentation. This is the first successful inter-species study on zebrafish by using blastula cell transplants from rosy barb both belonging to the same family cyprinidae.


Assessing ploidy levels and karyotype structure of the fire ant Solenopsissaevissima Smith, 1855 (Hymenoptera, Formicidae, Myrmicinae).
The family Formicidae is composed of ants that organize themselves into castes in which every individual has a joint organizational function. Solenopsis Westwood, 1840 is an ant genus with opportunistic and aggressive characteristics, known for being invasive species and stings that cause burning in humans. This genus is particularly difficult to classify and identify since its morphology provides few indications for species differentiation. For this, a tool that has been useful for evolutionary and taxonomic studies is cytogenetics. Here, we cytogenetically studied Solenopsissaevissima Smith, 1855 from Ouro Preto, Minas Gerais, Brazil. We evaluated the occurrence of polyploid cells in individuals and colonies by conventional cytogenetics. A total of 450 metaphases were analyzed and counted. Chromosome counts of individuals and colonies showed varied numbers of ploidies, from n = 16 to 8n = 128. The karyomorphometrical approach allowed determination of the following karyotypes: n = 10 m + 4 sm + 2 st, 2n = 20 m + 8 sm + 4 st, and 4n = 40 m + 16 sm + 8 st. Polyploidy can be found naturally in individuals and colonies and may represent an adaptative trait related to widespread distribution and invasion ability of new habitats.


Medicinal and injection therapies for mechanical neck disorders.
Medicinal therapies and injections are commonly recommended for neck pain, yet controversy persists over their effectiveness. To determine the effect of medicines and injections on pain, function/disability, patient satisfaction and range of motion in participants with mechanical neck disorders (MND). We searched CENTRAL (Issue 4, 2002), and MEDLINE, EMBASE, MANTIS, CINHAL from their start to March 2003. We scrutinized reference lists for other trials. We included randomized controlled trials with adults with MND, with or without associated headache or radicular findings. We considered medicinal and injection therapies, regardless of route of administration. Two authors independently selected articles, abstracted data and assessed methodological quality using the Jadad criteria. Consensus was used to resolve disagreements. When clinical heterogeneity was absent, we combined studies using random-effects meta-analysis models. We found 32 trials that examined the effects of oral NSAIDs, psychotropic agents, injections of steroids, and anaesthetic agents. Overall, methodological quality had a mean of 3.2/5 on the Jadad Scale. For acute whiplash, administering intravenous methylprednisolone within eight hours reduced pain at one week, and sick leave but not pain at six months compared to placebo. For chronic MND at short-term follow-up, intramuscular injection of lidocaine was superior to placebo or dry needling, but similar to ultrasound. In chronic MND with radicular findings, epidural methylprednisolone and lidocaine reduced neck pain and improved function at one-year follow-up compared to the intramuscular route. In subacute/chronic MND, we found conflicting evidence of pain reduction for oral psychotropic agents compared to placebo or control. Single trials of eperison hydrochloride and tetrazepam showed positive results. Results for cyclobenzaprine were mixed. Diazepam did not show benefit. Other treatments including NSAIDS and nerve blocks had unclear or limited evidence of benefit. In participants with chronic MND with or without radicular findings or headache, there was moderate evidence from five high quality trials showing that Botox A intramuscular injections were not better than saline in improving pain (pooled SMD: -0.39 (95%CI: -1.25 to 0.47), disability or global perceived effect. Intra-muscular injection of lidocaine for chronic MND and intravenous injection of methylprednisolone for acute whiplash were effective treatments. There was limited evidence of effectiveness of epidural injection of methylprednisolone and lidocaine for chronic MND with radicular findings. Oral psychotropic agents had mixed results. There was moderate evidence that Botox A intramuscular injections for chronic MND were no better than saline. Other medications, including NSAIDs, had contradictory or limited evidence of effectiveness.


Short communication: Processed bovine colostrum milk protein concentrate increases epithelial barrier integrity of Caco-2 cell layers.
Colostrum plays an important role in initiating the development of the intestinal barrier in newborn mammals. Given its bioactivity, there is much interest in the potential use of bovine colostrum to improve human gastrointestinal health throughout the life span. There is evidence that bovine colostrum is effective at improving small intestinal barrier integrity and some indication that it may alter colonic motility. However, for colostrum to be used as a product to improve intestinal health, it needs to be bioactive after processing. The aim of this study was to determine whether industrial processing of bovine colostrum affects its ability to improve small intestinal barrier integrity or alter distal colon motility. Three colostrum sample types were compared; raw whole colostrum powder (WCP), raw skim colostrum powder (SCP), and industrially produced colostrum milk protein concentrate (CMPC). To determine whether these colostrum powders had different effects on small intestinal barrier integrity, their effects on the transepithelial electrical resistance across an in vitro intestinal epithelial layer (Caco-2 cells) were measured, both with and without a challenge from the proinflammatory cytokine tumor necrosis factor-α. These results showed that CMPC enhanced transepithelial electrical resistance across unchallenged epithelial cell layers, whereas the raw colostrum samples, WCP and SCP, did not have an effect. The colostrum samples were also compared to determine how they affect contractility in the distal colon isolated from the rat. Skim colostrum powder was the only sample to act directly on colonic tissue to modulate motility, increasing the amplitude of contractions. The results show that bovine colostrum is able to improve small intestinal barrier integrity and alter colon motility, and they implicate different components. The barrier integrity enhancement was apparent only in the industrial CMPC, which may have been due to the increase in protein concentration or the release of small peptides as a result of processing. The ability to alter colon motility was present in SCP but absent in WCP, again implying that an increase in protein concentration is responsible for the effect. However, this effect was not apparent for the industrially processed CMPC, suggesting denaturation or degradation of the active component. The beneficial effect of colostrum on small intestinal barrier integrity was present after processing, confirming that it is feasible to industrially produce an active product for gut health.


Overexpression of miR-381-3p promotes the recovery of spinal cord injury.
To study the effects of miR-381-3p on spinal cord injury and its underlying mechanism. After the spinal cord injury rat model of was established, Sprague Dawley (SD) rats were randomly divided into the control group and the acute spinal cord injury (ASCI) group. Microglial BV2 cells were used as experimental cells, and the cells were divided into the control group and the lipopolysaccharide (LPS) group. The mRNA and protein expression level of miR-381-3p, IKKβ, inflammatory factors, and p-p65 were detected by quantitative Reverse Transcriptase-Polymerase Chain Reaction (qRT-PCR) and Western blot, respectively. Dual-luciferase reporter gene assay and Western blot were used to detect the regulatory effect of IKKβ on miR-381-3p. Changes in grip ability and rotary performance of rats in the ASCI group were evaluated after miR-381-3p overexpression in vivo. The expression of miR-381-3p was downregulated in rats of the ASCI group, while the expression of IKKβ and p-p65 were upregulated. In vitro experiments demonstrated that LPS could inhibit the expression of miR-381-3p and promote the upregulation of IKKβ and p-p65. Overexpression of miR-381-3p could inhibit the mRNA and protein expression of IKKβ. The upregulated expression of IKKβ, p-p65, tumor necrosis factor-alpha (TNF-α), and interleukins-1β (IL-1β) induced by LPS in BV2 cells were reversed by miR-381-3p mimic transfection. Besides, upregulated TNF-α and IL-1β induced by miR-381-3p inhibitor in BV2 cells were reversed by IKKβ inhibitor (BMS-345541). Results of animal experiments indicated that miR-381-3p was overexpressed in rats of the ASCI group. The protein levels of IKKβ and p-p65, and the mRNA expression levels of inflammatory cytokines TNF-α and IL-1β were remarkably decreased in the ASCI group than those of the control group. The grip ability, coordination, and anti-fatigue performance of rats in the ASCI group recovered quicker than those of the control group. MiR-381-3p was downregulated in ASCI rats. The overexpression of miR-381-3p could recover the motor ability of rats in the ASCI group earlier and might inhibit injury aggravation by inhibiting inflammatory responses via the IKKβ-NF-κB pathway.


Nutritional status of institutionalized elderly Brazilians: a study with the Mini Nutritional Assessment.
To assess the nutritional status of elderly living in nursing homes in the city of Salvador, Brazil and associated factors. Cross-sectional study performed with 359 individuals of both sexes, ages equal or over 60 years old, located in Nursing Homes in the urban area of the city of Salvador, Bahia, Brazil. Regarding nutritional status according to Mini Nutritional Assessment (MNA), 66.3% of the evaluated elderly were malnourished and at risk of malnutrition. When comparing sexes, it has been observed that among men the prevalence of this condition (76.6%) was higher than in women (62.4%). It has been observed, as a result of the multivariate analysis, that only the variable functional capacity for Activities of Daily Living (ADL) was statistically significant. There was moderate correlation between MNA and Mini-Mental State Examination (r=0.454; p<0.0001), as well as between MNA and the ADL scale (r=0.569; p<0.0001). There was weak negative correlation between MNA total score and age (r=0.159; p=0.002). Malnutrition and malnutrition risk were conditions of remarkable importance, with almost two-thirds of the elderly in this situation. ADL functional capacity must be monitored given their close relationship with the nutritional status of the elderly. An interdisciplinary approach in the context of institutionalization is needed due to the association between nutritional status and variables of different dimensions.


The effect of manual therapy on masseter muscle pain and spasm.
Orofacial pain and pain in the muscles of mastication are frequent symptoms of temporomandibular disorder. The masseter is the closet masticatory muscle to the surface and has the function of raising and retracting the mandible. This muscle has considerable strength and is one of the main muscles involved in the shredding of food It is therefore of utmost importance in the masticatory cycle and generally the most affected by pain and spasms. The aim of the present study was to analyze the effect of manual therapy with transversal and circular movements on pain and spasm in the masseter muscle, using electromyography and a visual analogue pain scale (VAPS). Eight women who experienced pain upon palpation of the masseter greater than 6 on the VAPS were selected for participation in the study, which employed electromyography and a VAPS for assessment, followed by manual oral physiotherapy and reevaluation. The statistical analysis revealed a reduction in pain, but there was no significant difference in electromyographic activity (p < 0.05). It was concluded that massage therapy was effective on pain symptoms, but was not capable of altering the electrical activity of the masseter muscle.


Patency of Individual and Sequential Coronary Artery Bypass in Patients with Ischemic Heart Disease: A Meta-analysis.
To evaluate the patency of individual and sequential coronary artery bypass in patients with ischemic heart disease. We searched PubMed, Cochrane Library, Excerpta Medica Database, and ClinicalTrials.gov databases for controlled trials. Endpoints included graft patency, anastomosis patency, occluded rates in left anterior descending (LAD) system and right coronary artery (RCA) system, in-hospital mortality, and follow-up mortality. Pooled risk ratios (RRs) and standardized mean difference (SMD) were used to assess the relative data. Nine cohorts, including 7100 patients and 1440 grafts under individual or sequential coronary artery bypass. There were no significant differences between individual and sequential coronary artery bypass in the graft patency (RR=0.96; 95% CI=0.91-1.02; P=0.16; I2=87%), anastomosis patency (RR=0.95; 95% CI=0.91-1.00; P=0.05; I2=70%), occluded rate in LAD system (RR=1.03; 95% CI=0.92-1.16; P=0.58; I2=37%), occluded rate in RCA system (RR=1.36; 95% CI=0.72-2.57; P=0.35; I2=95%), in-hospital mortality (RR=1.57; 95% CI=0.92-2.69; P=0.10; I2=0%), and follow-up mortality (RR=0.96; 95% CI=0.36-2.53; P=0.93; I2=0%). No significant differences on clinical data were observed regarding anastomosis patency, occluded rate in LAD system, occluded rate in RCA system, in-hospital mortality, and follow-up mortality, indicating that the patency of individual and the patency of sequential coronary artery bypass are similar to each other.


Bioluminescent methods for determining metabolites in micro-samples of pig plasma.
A highly sensitive and simplified method for the luminometric determination of plasma metabolites has been developed. Furthermore, the technique has been automated for the Dynatech ML2250 Microtiter Plate Luminometer and can be applied to the measurement of any plasma metabolite which may be coupled to a reaction involving the reduction of NAD+. Assays are described for lactose/galactose, beta-hydroxybutyrate and D-lactate, and have been validated with plasma samples. The assays require 1-2 microliters of plasma, and are capable of detecting concentrations below 5 mumol/l. Since luminometry is based on the kinetics of the luciferase/oxidoreductase enzyme system, components of complex biological samples may interfere with the rate of the reaction; necessitating the use of internal standards for individual samples. However, the need for internal standards to account for sample to sample variation in the luminescent response, has been eliminated with the present technique.


In vitro evaluation of the antibacterial potential and modification of antibiotic activity of the Eugenia uniflora L. essential oil in association with led lights.
Due to the great biodiversity of its flora, Brazil provides combat tools against bacterial resistance with the utilization of natural products with vegetable origin. Therefore, the present study had as its objective to evaluate the antibacterial potential of the Eugenia uniflora essential oil (EuEO) in vitro, as well as to analyze the modulatory effect of the oil against antibiotics by gaseous contact and to compare them when associated with a LED apparatus. The chemical components were characterised by gas chromatography which revealed the presence of the isoflurane-germacrene, considered the major component (61.69%). The MIC obtained from the EuEO was ≥256 μg/mL for S. aureus and ≥1024 μg/mL for E. coli. When combined with antibiotics, the EuEO presented synergism reducing the MIC when associated, with the exception of gentamicin against E. coli, where an antagonistic effect was observed. The was an interference of the EuEO over the activity of ciprofloxacin when associated with red and blue LED lights, increasing the inhibition halos against S. aureus and E. coli. Norfloxacin presented similar results to ciprofloxacin against S. aureus bacteria. When combined, norfloxacin and the EuEO presented synergism against S. aureus, which did not occur in the combination with ciprofloxacin. Interference occurred only with blue light for E. coli. Thus, it was observed that the EuEO causes changes in the activity of antibiotics, the same occurring with the use of LED lights, without significant differences in the association of the oil and the lights with the antibiotics tested. Further research is needed to elucidate the modulatory effects of the EuEO, as well as its association with LED lights.


CD46 expression in the central nervous system of male and female pubescent mice.
CD46 is a complementary regulatory protein ubiquitously expressed in human cells, controlling complement system activation. CD46 has further been identified to have several other functions including regulatory T cell induction and intestinal epithelial (IEC) barrier regulation. Activation of CD46 in the IEC can impact intestinal barrier permeability and immune system functioning. CD46 has only been identified in the spermatozoa and retina of mice. In other murine cells, the homologue CRRY is identified to function as the complementary regulator. Due to the identification of CRRY across other wild-type mouse cells and the development of mouse strains transgenic for human CD46, no recent research has been conducted to determine if CD46 is present in non-transgenic mouse strains. Therefore, the current study investigated if CD46 is expressed in the substantia nigra (SN) and caudate putamen (CP) of pubescent CD1 mice and examined the acute effects of pubertal antimicrobial and lipopolysaccharide (LPS) treatment on CD46 expression in the brain. As of 5 weeks of age, mice were administered mixed antimicrobial solution or water with oral gavage twice daily for 7 days. At 6 weeks of age, mice received an intraperitoneal injection of LPS or saline. Mice were euthanized 8 h post-injection and brain samples were collected. Our results indicate that pubescent CD-1 mice express CD46 in the SN and CP. However, LPS-treated mice displayed significantly less CD46 expression in the SN in comparison to saline-treated mice. Furthermore, males displayed more CD46 in the CP compared to females, regardless of LPS and antimicrobial treatments. Our data suggest CD46 is present in CD1 mice and that LPS and antimicrobial treatments impact CD46 protein expression in a sex-dependent manner. These results have important implications for the expression of CD46 in the mouse brain and the understanding of its role in immune system regulation.


Digital kids of the Heisei era: experiment at Toyonaka Bunka Kindergarten.
I wonder what most people think when they hear of small children playing on computers in kindergarten. One can almost hear the responses: "Why should kindergarten children have to use computers?" Small children are using computers? It's much too earlyl" "At kindergarten children should be singing, playing games, and making mud pies!" A computer is the epitome of the artificial. If we really make a mess of things, humans will become slaves to computers. What can they be thinking, putting such things in kindergartens as a child's toy? Many people who think in this way have never touched a computer, and it often seems the case that these "emotional opponents" are opposed to them only because of their own preconceptions. There is still a preconception that" a computer = a square machine like a TV with lots of difficult-looking keys". Computers are now in virtually all of the electronic appliances we use every day, refrigerators, washing machines, vacuums and televisions, and we think nothing of it. We live each day using computers, but only the "square" computer invites such contempt. Why is this the case? On the other side, there are the "proactive endorsers", who think "Computers have spread thus far in society and schools, so we must let children become familiar with them from a very young age!" These people often seem to want to teach everyone everything. There is much to know about the image and use of computers. They seem to think that if adults don't teach them, children won't understand anything. On this point, they are at the same level as the "emotional opponents", in that they conceive of computers as being something out of the ordinary.


Exploiting Preexisting Immunity to Enhance Oncolytic Cancer Immunotherapy.
Because of the high coverage of international vaccination programs, most people worldwide have been vaccinated against common pathogens, leading to acquired pathogen-specific immunity with a robust memory T-cell repertoire. Although CD8+ antitumor cytotoxic T lymphocytes (CTL) are the preferred effectors of cancer immunotherapy, CD4+ T-cell help is also required for an optimal antitumor immune response to occur. Hence, we investigated whether the pathogen-related CD4+ T-cell memory populations could be reengaged to support the CTLs, converting a weak primary antitumor immune response into a stronger secondary one. To this end, we used our PeptiCRAd technology that consists of an oncolytic adenovirus coated with MHC-I-restricted tumor-specific peptides and developed it further by introducing pathogen-specific MHC-II-restricted peptides. Mice preimmunized with tetanus vaccine were challenged with B16.OVA tumors and treated with the newly developed hybrid TT-OVA-PeptiCRAd containing both tetanus toxoid- and tumor-specific peptides. Treatment with the hybrid PeptiCRAd significantly enhanced antitumor efficacy and induced TT-specific, CD40 ligand-expressing CD4+ T helper cells and maturation of antigen-presenting cells. Importantly, this approach could be extended to naturally occurring tumor peptides (both tumor-associated antigens and neoantigens), as well as to other pathogens beyond tetanus, highlighting the usefulness of this technique to take full advantage of CD4+ memory T-cell repertoires when designing immunotherapeutic treatment regimens. Finally, the antitumor effect was even more prominent when combined with the immune checkpoint inhibitor anti-PD-1, strengthening the rationale behind combination therapy with oncolytic viruses. SIGNIFICANCE: These findings establish a novel technology that enhances oncolytic cancer immunotherapy by capitalizing on pre-acquired immunity to pathogens to convert a weak antitumor immune response into a much stronger one.


Differential expression and regulation of the c-src and c-fgr protooncogenes in myelomonocytic cells.
To study the expression of src-related protooncogenes during the development of myeloid cells and the regulation of these genes by the colony-stimulating factors that control myelopoiesis, normal monocytic cells at distinct stages of differentiation were derived from murine bone marrow with the monocytic lineage colony-stimulating factor CSF-1. Protooncogene expression was also examined in uncultured human myeloid leukemia cells. While c-src transcripts were detected in myeloid leukemia cells representative of all stages of differentiation, the highly related gene c-fgr was expressed at high levels only at later developmental stages, both in normal cells committed to the monocytic lineage and in leukemic cells with a differentiated myelomonocytic phenotype. When bone marrow-derived monocytic cells were synchronized and stimulated to proliferate with CSF-1, c-fgr transcripts (but not transcripts from the highly related genes c-src or c-yes) were induced 8 hr after the addition of CSF-1 and decreased to low levels by 20 hr as the monocytic cells entered S phase. The selective induction of c-fgr mRNA by CSF-1 suggests that this tyrosine kinase may have a unique function in normal monocytic cells, distinct from other src-related tyrosine kinases.


Assessment of autozygosity in Nellore cows (Bos indicus) through high-density SNP genotypes.
The use of relatively low numbers of sires in cattle breeding programs, particularly on those for carcass and weight traits in Nellore beef cattle (Bos indicus) in Brazil, has always raised concerns about inbreeding, which affects conservation of genetic resources and sustainability of this breed. Here, we investigated the distribution of autozygosity levels based on runs of homozygosity (ROH) in a sample of 1,278 Nellore cows, genotyped for over 777,000 SNPs. We found ROH segments larger than 10 Mb in over 70% of the samples, representing signatures most likely related to the recent massive use of few sires. However, the average genome coverage by ROH (>1 Mb) was lower than previously reported for other cattle breeds (4.58%). In spite of 99.98% of the SNPs being included within a ROH in at least one individual, only 19.37% of the markers were encompassed by common ROH, suggesting that the ongoing selection for weight, carcass and reproductive traits in this population is too recent to have produced selection signatures in the form of ROH. Three short-range highly prevalent ROH autosomal hotspots (occurring in over 50% of the samples) were observed, indicating candidate regions most likely under selection since before the foundation of Brazilian Nellore cattle. The putative signatures of selection on chromosomes 4, 7, and 12 may be involved in resistance to infectious diseases and fertility, and should be subject of future investigation.


Antibody response of captive cheetahs to modified-live feline virus vaccine.
The antibody response of cheetahs (Acinonyx jubatus) to modified live virus vaccine against feline panleukopenia (FPLV), herpes (FHV) and calici (FCV) viruses was assessed by means of an enzyme-linked immunosorbent assay (ELISA). In the first year of study, 82 cheetahs were bled pre-vaccination. Of these, antibody levels to FPLV were found in 100% of the animals. Only 54% were found to have antibodies to FHV and 99% had antibodies to FCV. One month after booster vaccination with the same vaccine, increased antibodies to FPLV, FHV and FCV were seen in 19 (58%), 18 (55%) and 25 (76%) of these animals, respectively (n = 33). In the second year of study, 65 cheetahs were bled pre-vaccination. Fifty three of these animals were negative for antibodies to FPLV while 28 were positive for FHV and 64 were positive for FCV. These animals were then bled 1, 2 and 6 mo post booster vaccination. The antibody levels to the various viruses showed different trends with time.


HIF-1alpha mRNA and protein upregulation involves Rho GTPase expression during hypoxia in renal cell carcinoma.
The small G proteins of the Rho family are involved in reorganization of the actin cytoskeleton, cell migration and in the regulation of gene transcription. Hypoxia-induced ATP depletion results in the disruption of actin organization which could affect Rho functions. In solid tumors, regions with low oxygen tension stimulate angiogenesis in order to increase oxygen and nutrient supply. This process is mediated by stabilization of the transcriptional factor hypoxia inducible factor 1 (HIF-1), which increases vascular endothelial growth factor (VEGF) production. In this study, we investigated the activities of Rho proteins, which are key regulators of cytoskeleton organization during hypoxia in renal cell carcinoma. Caki-1 cells were exposed to hypoxia (1% O2) and exhibited increased Cdc42, Rac1 and RhoA protein expression. Immunoprecipitation of metabolically labelled RhoA showed that overexpression was at least due to neo-synthesis. The Rho GTPases overexpressed during hypoxia were mainly located at membranes and pull-down assays demonstrated that they were active since they bound GTP. RT-PCR analysis indicated that the increase in RhoA protein expression was also reflected at the mRNA level. Overexpression and activation of Rho proteins were downstream of, and dependent on, the production of reactive oxygen species (ROS) since, in the presence of an inhibitor, both the rise of ROS and upregulation of Rho proteins were abolished. Importantly, preincubation of cells with the toxin C3, which inhibits RhoA, reduced HIF-1alpha protein accumulation by 84% during hypoxia. Together, these results support a model where ROS upregulate Rho protein expression and where active RhoA is required for HIF-1alpha accumulation during hypoxia.


Altered behavioural responses and functional recovery in rats following sciatic nerve compression and early vs late decompression.
The aim of the study was to examine sensory behaviour and functional recovery in rats during nerve compression and after decompression. Compression injury is a far more common condition than nerve transection. The condition is characterised by numbness and a tingling/burning sensation, and some patients experience pain and allodynia during compression or after decompression treatment. The aetiology is in many cases unknown. Thus, further studies are of great importance for the understanding of this condition. In the present study, behavioural responses to tactile stimulation, thermal pain, as well as functional sensorimotor behaviour were investigated in rats before, during severe compression, and after decompression. The sciatic nerve of the rats was experimentally compressed for 3 or 28 days, whereafter surgical release, i.e. decompression, of the nerve was performed and the rats were examined up to ∼9 weeks. An altered behaviour was found in response to compression injury, which is mitigated after early decompression treatment. These findings indicate that early intervention during severe compression injuries is of great importance for recovery and restoration of nerve function and, thus, should have an impact on clinical routines regarding treatment of compression injuries.


Application of sulfobutylether-beta-cyclodextrin with specific degrees of substitution for the enantioseparation of pharmaceutical mixtures by capillary electrophoresis.
In this research the separation of the enantiomers of the basic drug bidisomide (SC-40230) from five closely related known process impurities was investigated using several neutral and anionic sulfobutylether beta-cyclodextrins (SBE-beta-CDs) as isomer selectors. Several novel sulfobutylether derivative mixtures and purified charge types having a specific degree of substitution were used to study the effect of selector charge on the efficiency and selectivity of both chiral and achiral separations. The effects of run buffer pH, selector type, and selector concentration on the chiral separation of bidisomide and the achiral separation of the related process impurities was also investigated. The related process impurity, SC-47500, displayed significant peak tailing with SBE-beta-CD mixtures which contained mono- to deca-substituted cyclodextrins. This problem was explored using isolated SBE-beta-CD charge types having degrees of substitution from one to seven. Peak tailing increased as the charge on the selector increased, suggesting that the distortion was due to electrodispersion and the large countercurrent mobility of the negatively charged complexes. Pure charge types having a lower degree of substitution provided adequate chiral and achiral selectivity, while eliminating the severe peak distortion caused by electrodispersion. The complete analysis of the bidisomide enantiomers and the related impurities was achieved with a pH 2.5 running buffer containing 5-10 mM of the isolated sulfobutylether charge types SBE[2]ds(1)sr-beta-CD or SBE[3]ds(1)sr-beta-CD. These conditions gave baseline resolution of bidisomide enantiomers and all five impurities, thus allowing both chiral and achiral purity to be determined in a single run.


Coat colour in mouse populations selected for weight gain: support for hitchhiking, not pleiotropy.
With many molecular markers in many species, research efforts in quantitative genetics have focused on dissecting these traits and understanding the importance of factors such as correlated response due to hitchhiking or pleiotropy. Here, in an examination of long-term selection experiments in mice, the evidence strongly supports the primary importance of hitchhiking on the coat colour loci brown and dilute in mice selected for high weight gain. First, the amount of observed change in coat colour allele frequency could not be explained by genetic drift alone, implying that selection was of high importance. Second, the allele frequency changes included reversals in the direction change, but there were still positive correlations in the early generations with differences in weight gain between the phenotypes. Third, the correlation between the change in allele frequencies and phenotypic difference in weight gain declined over time, consistent with the decay expected from linkage associations. Fourth, the changes at both loci in a short-term selection experiment for low weight gain were in the opposite direction than the changes in the contemporaneous related population selected for high weight gain.


Role of chloroplast trienoic fatty acids in plant disease defense responses.
Trienoic fatty acids (TAs) are the major polyunsaturated fatty acid species in the membrane lipids in plant cells. TAs are crucial for the adaptation to abiotic stresses, especially low- or high-temperature stress. We show that TAs in chloroplast membrane lipids are involved in defense responses against avirulent bacterial pathogens. Avirulent pathogen invasion of plants induces a transient production of reactive oxygen intermediates (ROI), programmed cell death and subsequent disease resistance. The Arabidopsis fad7fad8 mutation, which prevents the synthesis of TAs in chloroplast lipids, caused the reduction in ROI accumulation in leaves inoculated with Pseudomonas syringae pv. tomato DC3000 (avrRpm1). Linolenic acid, the most abundant TA, activated the NADPH oxidase that is responsible for ROI generation. TAs were transferred from chloroplast lipids to extrachloroplast lipids coincident with ROI accumulation after inoculation with Pst DC3000 (avrRpm1). Furthermore, the fad7fad8 mutant exhibited reduced cell death and was compromised in its resistance to several avirulent P. syringae strains. These results suggest that TAs derived from chloroplast lipids play an important role in the regulation of plant defense responses.


The hydrolysis of algal galactans by enzymes from a Cytophaga species.
1. Two bacteria were isolated from sea water by the enrichment culture technique, both of which could utilize the galactan sulphate, porphyran, as sole source of carbon. 2. From the cells of one bacterium, classified as a Cytophaga sp., hydrolytic enzymes were isolated. 3. Partial purification of the enzymes is described and some of the properties of the principal enzymes have been studied. 4. The action of the enzymes on several galactan sulphates of red algae suggests that an agarase is present in the mixture.


Recombinant human liver betaine-homocysteine S-methyltransferase: identification of three cysteine residues critical for zinc binding.
Betaine-homocysteine S-methyltransferase (BHMT; EC 22.214.171.124) catalyzes the transfer of an N-methyl group from betaine to homocysteine to produce dimethylglycine and methionine, respectively. The enzyme is found in the pathway of choline oxidation and is abundantly expressed in liver and kidney. We have recently shown that human BHMT is a zinc metalloenzyme [Millian, N. S., and Garrow, T. A. (1998) Arch. Biochem. Biophys. 356, 93-98]. To facilitate the rapid purification of human BHMT for further physical and mechanistic studies, including characterizing its metal binding properties, we have overexpressed the enzyme in E. coli as a fusion construct which facilitated its subsequent purification by a self-cleavable affinity tag system (IMPACT T7). Using this expression and purification system in conjunction with site-directed mutagenesis, we have identified Cys217, Cys299, and Cys300 as zinc ligands. Mutating any of these Cys residues to Ala results in the complete loss of activity and a significant reduction in the ability of the protein to bind zinc. Comparing the regions of BHMT amino acid sequence surrounding these Cys residues with similar amino acid sequences retrievable from protein databases, we have identified the following motif: G[ILV]NCX(20,100)[ALV]X(2)[ILV]GGCCX(3)PX(2)I, which we propose to be a signature for a family of zinc-dependent methyltransferases that utilize thiols or selenols as methyl acceptors. Some of the members of this family include the vitamin B(12)-dependent methionine synthases, E. coli S-methylmethionine-S-homocysteine methyltransferase, and A. bisulcatus S-methylmethionine-selenocysteine methyltransferase.


The Role of Radiotherapy in Orbital Pseudotumor: A Systematic Review of Literature.
Orbital pseudotumor (or pseudotumor orbitae) is a benign entity, also known as idiopathic orbital inflammatory syndrome (IOIS), which encompasses a wide range of non-tumoral, space occupying lesions of the orbit. In selected cases of refractory disease or presence of side effects or even comorbidities that limit systemic therapies, radiotherapy could play a role in the management because it has been demonstrated that irradiation is effective in suppressing the inflammatory process. A systematic review of the literature about the main scientific databases was launched and the time interval included all published articles present in the databases from their inception until September 2020. We were able to identify 19 studies eligible for inclusion in this review from 1978 to 2018. Overall the data of 241 patients were collected and are presented in this systematic review. The response rate varied between 74% and 100% with a median recurrence rate of 10%. The median total dose was 20 Gy whereas the mean total dose was 21 Gy with a range from 4 to 36 Gy. Regarding the fractionation, 2 Gy/fraction daily was the most widely used. Radiotherapy seemed to achieve good response rates however, in most of the studies, inclusion criteria and outcome parameters are not uniform and therefore the results are difficult to compare. Often important parameters such as chronic pain and permanent functional deficits are not assessed in the outcome. Therefore, prospective studies, with good cohort characteristics and a clear definition of the outcome, are required.


[A randomized study of efficacy of regimes of large-fraction irradiation for metastatic lesions to the skeleton. (I) Single factor analysis].
A randomized study included 167 patients with metastases to the bone (breast cancer--73%). 117 patients received distance radiotherapy on an area-by-area basis: 2 areas--30, and 3 or more areas--20. Radiotherapy was not used until chemo-hormonal proved useless. The following four regimes were used (1) 4 fractions of 6.5 Gy each (overall dosage--26 Gy) (2 fractions per week); (2) 5 fractions of 6.5 Gy each (overall dosage--32.5 Gy) (2 fractions per week); (3) 4 daily fractions of 6.5 Gy each (overall dosage--26 Gy), and (4) 23 fractions (overall dosage--46 Gy) (5 weekly fractions). Overall efficacy was 96.7% (complete response--66.7%; pain syndrome in the exposed area--4.2%). No differences between large-fraction irradiation regimes were reported. Late-onset radiation-induced lesions (soft-tissue fibrosis stage II-III) were significantly more frequent after standard low-dosage fractionated irradiation (28.6+/-17.0%).


Pneumomediastinum in a female track and field athlete: a case report.
To present the case of an elite female track and field athlete who suffered a pneumomediastinum resulting from a Valsalva maneuver performed while throwing the javelin. Episodes of chest pain and labored breathing in athletes may be alarming. Accurate, early diagnosis is enhanced by an awareness of those relatively rare conditions that may cause these symptoms. Bronchial injury/fracture, retropharyngeal abscess, acute pulmonary disease, pneumomediastinum, pneumothorax, cardiac disease, allergic reaction. The athlete was given intravenous morphine for pain, prescribed oral pain medication, and restricted from strenuous activity for 6 weeks. Aerobic exercise was allowed after pain and air in the neck subsided, which was estimated at 1 week postinjury. This is a rarely reported case of a pneumomediastinum in a female and a track and field athlete. Medical personnel must be aware of the possibility of pneumomediastinum in track and field athletes and in female athletes and must be knowledgeable in the followup care and the safe return of the athlete to activity.


[Experimental study, using a pulmonary model, of the pathogenic properties of genetically related strains of Sh. flexneri, differing in their ability to cause keratoconjunctivitis].
Complex microbiological and morphological study of the genetically connected Sh. flexneri strains, avirulent by keratoconjunctival test showed that the greatest extent of the virulent loss was revealed in shigellae with mutation in the area glpK-gene: they lost the capacity to penetrate into the cells of the bronchial epithelium, to resist local leukocytic reaction and to produce an injurious action on the pulmonary vessels. Shigellae with the replaced kcpA- gene area mostly lost the capacity to penetrate into the epithelial cells of mouse bronchi and were eliminated from the epithelium by the 9th hour after the administration. Xyl--avir-hybrids obtained from crossing of Sh flexneri with the streptomycin-resistant E. coli K12 donors were characterized by the loss of the capacity to prolonged reproduction in the organism of mice with the retention of the capacity to penetration into the epithelium, of the influence on the development of the leukocytic reaction and of the damaging effect on the pulmonary capillaries.


Evidence for a qualitative hierarchy within the Hoechst-33342 'side population' (SP) of murine bone marrow cells.
In vitro cobblestone area (CA)-forming cell (CAFC) and in vivo (short-term and competitive repopulation) assays demonstrate that a qualitative hierarchy exists within the Hoechst-33342-defined side population (SP) in murine bone marrow (BM). Consistent with and extending previous studies, we demonstrate that (i) hematopoietic activity found in whole BM (WBM) is concentrated within the SP, rather than the non-SP (NSP); and (ii) within the SP, those cells that more strongly efflux the dye (lower SP, LSP) are qualitatively different from those that less strongly efflux the dye (upper SP, USP). Qualitative differences are highlighted by evidence that (i) CA derived from LSP CAFC persist in culture significantly longer than CA derived from USP CAFC; (ii) short-term, multilineage repopulation of lethally irradiated mice by LSP cells is more rapid than that in mice receiving USP, NSP, whole SP (WSP), or WBM cells and (iii) LSP cells out-compete USP cells in the multilineage hematopoietic repopulation of lethally irradiated recipients. These data suggest that LSP cells are of higher quality than USP cells and potentially provide a means by which qualitative changes in primitive hematopoietic progenitors occurring naturally with aging, or clinically as a consequence of therapeutic manipulation, can be assessed.


The chemokine receptor CCR4 promotes tumor growth and lung metastasis in breast cancer.
Increasing evidence has shown that chemokines and chemokine receptors are associated with tumor growth and metastasis. CCR4, an important chemokine receptor for regulating immune homeostasis, is thought to be involved in hematologic malignancies and has also recently implicated in some solid tumors, such as gastric cancer. The possible role of CCR4 in breast cancer has not been well elucidated. In this study, we show that CCR4 is differentially expressed in human breast cancer cell lines. Specifically, we find that CCR4 is overexpressed in breast cancer cell lines with high metastatic potential. More importantly, we used a combination of overexpression and RNA interference to demonstrate that CCR4 promotes breast tumor growth and lung metastasis in mice. Furthermore, we find that microvessel density is significantly increased in tumors formed by CCR4-overexpressing cells and decreased in those formed by CCR4-knockdown cells. We find that overexpression of CCR4 can enhance the chemotactic response of breast cancer cells to CCL17. However, the expression of CCR4 does not affect the proliferation of breast cancer cells in vitro. Furthermore, we show that CCR4 expression is positively correlated with HER2 expression, tumor recurrence and lymph node, lung and bone metastasis (P < 0.05). Multivariate analysis showed that CCR4 expression is a significant independent prognostic factor for overall survival (P = 0.036) but not for disease-free survival in patients with breast cancer (P = 0.071). Survival analysis indicated a strong association between CCR4 expression and lower overall survival (P = 0.0001) and disease-free survival (P = 0.016) in breast cancer.


MUC4 potentiates invasion and metastasis of pancreatic cancer cells through stabilization of fibroblast growth factor receptor 1.
MUC4 is a type-1 transmembrane mucin differentially expressed in multiple cancers and has previously been shown to potentiate progression and metastasis of pancreatic cancer. In this study, we investigated the molecular mechanisms associated with the MUC4-induced invasion and metastasis in pancreatic cancer. Stable silencing of MUC4 in multiple pancreatic cancer cells resulted in the downregulation of N-cadherin and its interacting partner fibroblast growth factor receptor 1 (FGFR1) through downregulation of partly by pFAK, pMKK7, pJNK and pc-Jun pathway and partly through PI-3K/Akt pathway. The downregulation of FGFR1 in turn led to downregulation of pAkt, pERK1/2, pNF-κB, pIkBα, uPA, MMP-9, vimentin, N-cadherin, Twist, Slug and Zeb1 and upregulation of E-cadherin, Occludin, Cytokeratin-18 and Caspase-9 in MUC4 knockdown BXPC3 and Capan1 cells compared with scramble vector transfected cells. Further, downregulation of FGFR1 was associated with a significant change in morphology and reorganization of the actin-cytoskeleton, leading to a significant decrease in motility (P < 0.00001) and invasion (P < 0.0001) in vitro and decreased tumorigenicity and incidence of metastasis in vivo upon orthotopic implantation in the athymic mice. Taken together, the results of the present study suggest that MUC4 promotes invasion and metastasis by FGFR1 stabilization through the N-cadherin upregulation.


Comparative effects between electronic cigarette and tobacco cigarette smoke on oxidative markers in cultured immune cells isolated from rats.
Tobacco cigarette smoke (TCS) was previously demonstrated to affect the innate and adaptive immune responses as a consequence of oxidant generation which play a pivotal role in neutrophilic airway inflammation. Aim of this paper was to investigate whether electronic cigarette smoke (ECS) generates reactive oxygen species (ROS) similarly to cigarette smoke. By means of a house made apparatus, ECS and TCS were collected in fetal bovine serum (FBS) which was used to grow immune cells isolated from rats. As index of oxidative products nitrite, superoxide, and thiobarbituric acid-reactive substances (TBARS) were determined in the medium before and after cell growth. The results showed that: i) ECS caused a remarkable increase of nitrites and TBARS although in lesser extension than TCS; ii) the spleen and lymph node cells grown in ECS and TCS-exposed medium were able to reduce TBARS but not nitrites present in the medium; iii) PBMC in TCS-exposed medium were able to reduce nitrites and TBARS more efficiently than spleen and lymph node cells, but released more superoxide anion; iv) TCS and ECS not influence the PBMC and spleen T cell subtype populations (CD4+, CD8+). As ECS nicotine-free gave the same results of unexposed medium, we can support the hypothesis that the increase of ROS in ECS exposed medium was prevalently due to nicotine.


MAVS self-association mediates antiviral innate immune signaling.
The innate immune system recognizes nucleic acids during viral infection and stimulates cellular antiviral responses. Intracellular detection of RNA virus infection is mediated by the RNA helicases RIG-I (retinoic acid inducible gene I) and MDA-5, which recognize viral RNA and signal through the adaptor molecule MAVS (mitochondrial antiviral signaling) to stimulate the phosphorylation and activation of the transcription factors IRF3 (interferon regulatory factor 3) and IRF7. Once activated, IRF3 and IRF7 turn on the expression of type I interferons, such as beta interferon. Interestingly, unlike other signaling molecules identified in this pathway, MAVS contains a C-terminal transmembrane (TM) domain that is essential for both type I interferon induction and localization of MAVS to the mitochondrial outer membrane. However, the role the MAVS TM domain plays in signaling remains unclear. Here we report the identification of a function for the TM domain in mediating MAVS self-association. The activation of RIG-I/MDA-5 leads to the TM-dependent dimerization of the MAVS N-terminal caspase recruitment domain, thereby providing an interface for direct binding to and activation of the downstream effector TRAF3 (tumor necrosis factor receptor-associated factor 3). Our results reveal a role for MAVS self-association in antiviral innate immunity signaling and provide a molecular mechanism for downstream signal transduction.