Exploring microbiota-gut-brain axis biomarkers linked to autism spectrum disorder in prenatally chlorpyrifos-exposed Fmr1 knock-out and wild-type male rats.

Fmr1 (fragile X messenger ribonucleoprotein 1)-knockout (KO) rats, modeling the human Fragile X Syndrome (FXS), are of particular interest for exploring the ASD-like phenotype in preclinical studies. Gestational exposure to chlorpyrifos (CPF) has been associated with ASD diagnosis in humans and ASD-like behaviors in rodents and linked to the microbiota-gut-brain axis. In this study, we have used both Fmr1-KO and wild-type male rats (F2 generation) at postnatal days (PND) 7 and 40 obtained after F1 pregnant females were randomly exposed to 1 mg/kg/mL/day of CPF or vehicle. A nuclear magnetic resonance (NMR) metabolomics approach together with gene expression profiles of these F2 generation rats were employed to analyze different brain regions (such as prefrontal cortex, hippocampus, and cerebellum), whole large intestine (at PND7) and gut content (PND40). The statistical comparison of each matrix spectral profile unveiled tissue-specific metabolic fingerprints. Significant variations in some biomarker levels were detected among brain tissues of different genotypes, including taurine, myo-inositol, and 3-hydroxybutyric acid, and exposure to CPF induced distinct metabolic alterations, particularly in serine and myo-inositol. Additionally, this study provides a set of metabolites associated with gastrointestinal dysfunction in ASD, encompassing several amino acids, choline-derived compounds, bile acids, and sterol molecules. In terms of gene expression, genotype and gestational exposure to CPF had only minimal effects on decarboxylase 2 (gad2) and cholinergic receptor muscarinic 2 (chrm2) genes.

NMR-based Metabolomics and Fatty Acid Profiles to Unravel Biomarkers in Preclinical Animal Models of Compulsive Behavior.

Compulsivity is a key manifestation of inhibitory control deficit and a cardinal symptom of psychopathological conditions such as obsessive-compulsive and attention-deficit hyperactivity disorders, in which metabolic alterations have raised attention as putative biomarkers for early identification. The present study assessed the metabolic profile in a preclinical model of a compulsive phenotype of rats. We used the schedule-induced polydipsia (SIP) method to classify male Wistar rats into high drinkers (HDs) or low drinkers (LDs) according to their compulsive drinking rate developed by exposure to a fixed-time 60 s (FT-60) schedule of reinforcement with water available ad libitum during 20 sessions. Before and after SIP, blood samples were collected for subsequent serum analysis by nuclear magnetic resonance spectroscopy coupled to multivariate analysis. Although no differences existed in the pre-SIP set, the compulsive drinking behavior induced remarkable metabolic alterations: HD rats selected by SIP exhibited a hyperlipidemic, hypoglycemic, and hyperglutaminergic profile compared with their low-compulsive counterparts. Interestingly, these alterations were not attributable to the mere exposure to reward pellets because a control experiment did not show differences between HDs and LDs after 20 sessions of pellet consumption without intermittent reinforcement. Our results shed light toward the implication of dietary and metabolic factors underpinning the vulnerability to compulsive behaviors.

RETINAL EMBOLIZATION AFTER CAROTID ECHO DOPPLER: A CASE REPORT.

PURPOSE: To report a case of symptomatic multiple retinal emboli during a carotid Doppler ultrasonography (CDU) procedure. METHODS: Case report documented with clinical records, fundus photography, optical coherence tomography angiography of the retina, and computed tomography angiography of the supraaortic vessels. RESULTS: A 60-year-old male patient presented to the emergency department with sudden vision loss in the left eye and pain in the left upper eyelid noticed during a CDU procedure. On the left eye, the best-corrected visual acuity was 20/200, and fundus observation revealed retinal arteriolar attenuation, retinal whitening with cherry point, and multiple intraarterial emboli in all arcades of the central retinal artery. Computed tomography angiography of the supraaortic vessels revealed an absence of filling throughout the cervical, petrous, and cavernous segment of the left internal carotid artery, resuming only in the ophthalmic segment and an important stenosis of the left external carotid artery. He had no indication to be submitted to left carotid endarterectomy and began ocular massage and oral anticoagulant. Four months later, ptosis was no longer present, best-corrected visual acuity improved to 20/20 with a residual scotoma, and emboli disappeared on fundus photography. CONCLUSION: Embolic occlusion of retinal arteries may be an extremely rare but serious complication associated with CDU, and patients and clinicians should be aware of it.

Development and Validation of an Instrument to Assess Horseback Mobility in Hippotherapy.

Improvement in rider mobility represents an important functional gain for people with disabilities undergoing hippotherapy. However, there is no validated measuring instrument to track and document the rider's progress in riding activities. In this study, we aimed to develop and establish validity evidence for an instrument to assess hippotherapy participants' mobility on horseback. We report on this development through the stages of: (a) content validation, (b) construct validation, (c) inter- and intra-rater reliability and (d) internal consistency analysis. We evaluated its factor structure with exploratory factor analyses, calculated values for inter- and intra-rater reliability using the intra-class correlation coefficient, and calculated its internal consistency using Cronbach's alpha. We followed recommendations by the Guidelines for Reporting Reliability and Agreement Studies. We found good inter-rater reliability (intra-class correlation coefficient - ICC = 0.991-0.999) and good intra-rater reliability (ICC = 0.997-1.0), and there was excellent internal consistency (Cronbach's α = 0.937-0.999). The instrument's factor structure grouped its three domains into one factor. As this instrument is theoretically consistent and has been found to be appropriate and reliable for its intended use, it is now available for the measurement of horseback mobility among hippotherapy riders.

Dissonant health transition in the states of Mexico, 1990-2013: a systematic analysis for the Global Burden of Disease Study 2013.

BACKGROUND: Child and maternal health outcomes have notably improved in Mexico since 1990, whereas rising adult mortality rates defy traditional epidemiological transition models in which decreased death rates occur across all ages. These trends suggest Mexico is experiencing a more complex, dissonant health transition than historically observed. Enduring inequalities between states further emphasise the need for more detailed health assessments over time. The Global Burden of Diseases, Injuries, and Risk Factors Study 2013 (GBD 2013) provides the comprehensive, comparable framework through which such national and subnational analyses can occur. This study offers a state-level quantification of disease burden and risk factor attribution in Mexico for the first time. METHODS: We extracted data from GBD 2013 to assess mortality, causes of death, years of life lost (YLLs), years lived with disability (YLDs), disability-adjusted life-years (DALYs), and healthy life expectancy (HALE) in Mexico and its 32 states, along with eight comparator countries in the Americas. States were grouped by Marginalisation Index scores to compare subnational burden along a socioeconomic dimension. We split extracted data by state and applied GBD methods to generate estimates of burden, and attributable burden due to behavioural, metabolic, and environmental or occupational risks. We present results for 306 causes, 2337 sequelae, and 79 risk factors. FINDINGS: From 1990 to 2013, life expectancy from birth in Mexico increased by 3·4 years (95% uncertainty interval 3·1-3·8), from 72·1 years (71·8-72·3) to 75·5 years (75·3-75·7), and these gains were more pronounced in states with high marginalisation. Nationally, age-standardised death rates fell 13·3% (11·9-14·6%) since 1990, but state-level reductions for all-cause mortality varied and gaps between life expectancy and years lived in full health, as measured by HALE, widened in several states. Progress in women's life expectancy exceeded that of men, in whom negligible improvements were observed since 2000. For many states, this trend corresponded with rising YLL rates from interpersonal violence and chronic kidney disease. Nationally, age-standardised YLL rates for diarrhoeal diseases and protein-energy malnutrition markedly decreased, ranking Mexico well above comparator countries. However, amid Mexico's progress against communicable diseases, chronic kidney disease burden rapidly climbed, with age-standardised YLL and DALY rates increasing more than 130% by 2013. For women, DALY rates from breast cancer also increased since 1990, rising 12·1% (4·6-23·1%). In 2013, the leading five causes of DALYs were diabetes, ischaemic heart disease, chronic kidney disease, low back and neck pain, and depressive disorders; the latter three were not among the leading five causes in 1990, further underscoring Mexico's rapid epidemiological transition. Leading risk factors for disease burden in 1990, such as undernutrition, were replaced by high fasting plasma glucose and high body-mass index by 2013. Attributable burden due to dietary risks also increased, accounting for more than 10% of DALYs in 2013. INTERPRETATION: Mexico achieved sizeable reductions in burden due to several causes, such as diarrhoeal diseases, and risks factors, such as undernutrition and poor sanitation, which were mainly associated with maternal and child health interventions. Yet rising adult mortality rates from chronic kidney disease, diabetes, cirrhosis, and, since 2000, interpersonal violence drove deteriorating health outcomes, particularly in men. Although state inequalities from communicable diseases narrowed over time, non-communicable diseases and injury burdens varied markedly at local levels. The dissonance with which Mexico and its 32 states are experiencing epidemiological transitions might strain health-system responsiveness and performance, which stresses the importance of timely, evidence-informed health policies and programmes linked to the health needs of each state. FUNDING: Bill & Melinda Gates Foundation, Instituto Nacional de Salud Pública.

Behaviours and opinions towards outdoor smoking bans and cigarette littering in Baja California, Mexico.

OBJECTIVE: Outdoor smoking bans have been implemented to reduce secondhand smoke exposure and cigarette littering. We aimed to explore outdoor smoking behaviours, cigarette littering, and support towards a recently enacted outdoor smoking ban in Baja California, Mexico. METHODS: From October 2013 to April 2014 a representative sample of 800 adults from urban areas of Baja California was surveyed to obtain information on demographics, smoking status, opinion towards outdoor smoking bans in general and in specific locations, and perceptions and habits regarding cigarette littering. Descriptive statistics and Poisson regression analyses were conducted to estimate prevalence ratios and corresponding 95% CI taking into account the complex survey design. RESULTS: Outdoor smoking was frequently observed, as 86.8% of the population reported having seen people smoking in parks and beaches and 94.4% in open venues such as concerts or stadiums. A smoking ban in all outdoor areas was supported by 71.8% of Baja California residents. Site-specific support ranged from 80% (stadiums) to 89% (public parks). Support for the ban was higher among never and ex-smokers than in current smokers. Cigarette littering was negatively perceived by 98% of the participants, yet 45% of all cigarettes smoked were thrown on the ground. Based on cigarette consumption and butt littering reported in the survey, we estimated that 27 million cigarette butts become litter every month in urban areas of Baja California. CONCLUSIONS: Outdoor smoking is still frequent in outdoor areas where people gather, and an important source of littering. We found substantial public support towards the outdoor smoking ban, which should translate into easier implementation and compliance in Baja California, Mexico.

Antibacterial activity and mode of action of selected glucosinolate hydrolysis products against bacterial pathogens.

Plants contain numerous components that are important sources of new bioactive molecules with antimicrobial properties. Isothiocyanates (ITCs) are plant secondary metabolites found in cruciferous vegetables that are arising as promising antimicrobial agents in food industry. The aim of this study was to assess the antibacterial activity of two isothiocyanates (ITCs), allylisothiocyanate (AITC) and 2-phenylethylisothiocyanate (PEITC) against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Listeria monocytogenes. The antibacterial mode of action was also characterized by the assessment of different physiological indices: membrane integrity, intracellular potassium release, physicochemical surface properties and surface charge. The minimum inhibitory concentration (MIC) of AITC and PEITC was 100 µg/mL for all bacteria. The minimum bactericidal concentration (MBC) of the ITCs was at least 10 times higher than the MIC. Both AITC and PEITC changed the membrane properties of the bacteria decreasing their surface charge and compromising the integrity of the cytoplasmatic membrane with consequent potassium leakage and propidium iodide uptake. The surface hydrophobicity was also non-specifically altered (E. coli and L. monocytogenes become less hydrophilic; P. aeruginosa and S. aureus become more hydrophilic). This study shows that AITC and PEITC have strong antimicrobial potential against the bacteria tested, through the disruption of the bacterial cell membranes. Moreover, phytochemicals are highlighted as a valuable sustainable source of new bioactive products.

Antimicrobial Activity of Selected Phytochemicals against Escherichia coli and Staphylococcus aureus and Their Biofilms.

Bacteria can be resistant to multiple antibiotics and we are fast approaching a time when antibiotics will not work on some bacterial infections. New antimicrobial compounds are urgently necessary. Plants are considered the greatest source to obtain new antimicrobials. This study aimed to assess the antimicrobial activity of four phytochemicals-7-hydroxycoumarin (7-HC), indole-3-carbinol (I3C), salicylic acid (SA) and saponin (SP)-against Escherichia coli and Staphylococcus aureus, either as planktonic cells or as biofilms. These bacteria are commonly found in hospital-acquired infections. Some aspects on the phytochemicals mode of action, including surface charge, hydrophobicity, motility and quorum-sensing inhibition (QSI) were investigated. In addition, the phytochemicals were combined with three antibiotics in order to assess any synergistic effect. 7-HC and I3C were the most effective phytochemicals against E. coli and S. aureus. Both phytochemicals affected the motility and quorum-sensing (QS) activity, which means that they can play an important role in the interference of cell-cell interactions and in biofilm formation and control. However, total biofilm removal was not achieved with any of the selected phytochemicals. Dual combinations between tetracycline (TET), erythromycin (ERY) and ciprofloxacin (CIP) and I3C produced synergistic effects against S. aureus resistant strains. The overall results demonstrates the potential of phytochemicals to control the growth of E. coli and S. aureus in both planktonic and biofilm states. In addition, the phytochemicals demonstrated the potential to act synergistically with antibiotics, contributing to the recycling of old antibiotics that were once considered ineffective due to resistance problems.

Current and emergent strategies for disinfection of hospital environments.

A significant number of hospital-acquired infections occur due to inefficient disinfection of hospital surfaces, instruments and rooms. The emergence and wide spread of multiresistant forms of several microorganisms has led to a situation where few compounds are able to inhibit or kill the infectious agents. Several strategies to disinfect both clinical equipment and the environment are available, often involving the use of antimicrobial chemicals. More recently, investigations into gas plasma, antimicrobial surfaces and vapour systems have gained interest as promising alternatives to conventional disinfectants. This review provides updated information on the current and emergent disinfection strategies for clinical environments.

Persister cells in a biofilm treated with a biocide.

This study investigated the physiology and behaviour following treatment with ortho-phthalaldehyde (OPA), of Pseudomonas fluorescens in both the planktonic and sessile states. Steady-state biofilms and planktonic cells were collected from a bioreactor and their extracellular polymeric substances (EPS) were extracted using a method that did not destroy the cells. Cell structure and physiology after EPS extraction were compared in terms of respiratory activity, morphology, cell protein and polysaccharide content, and expression of the outer membrane proteins (OMP). Significant differences were found between the physiological parameters analysed. Planktonic cells were more metabolically active, and contained greater amounts of proteins and polysaccharides than biofilm cells. Moreover, biofilm formation promoted the expression of distinct OMP. Additional experiments were performed with cells after EPS extraction in order to compare the susceptibility of planktonic and biofilm cells to OPA. Cells were completely inactivated after exposure to the biocide (minimum bactericidal concentration, MBC = 0.55 ± 0.20 mM for planktonic cells; MBC = 1.7 ± 0.30 mM for biofilm cells). After treatment, the potential of inactivated cells to recover from antimicrobial exposure was evaluated over time. Planktonic cells remained inactive over 48 h while cells from biofilms recovered 24 h after exposure to OPA, and the number of viable and culturable cells increased over time. The MBC of the recovered biofilm cells after a second exposure to OPA was 0.58 ± 0.40 mM, a concentration similar to the MBC of planktonic cells. This study demonstrates that persister cells may survive in biocide-treated biofilms, even in the absence of EPS.

Ressincronização cardíaca e terapia celular: existe terapia associativa? / Cardiac resynchronization and stem cell therapy: do associative benefits exist?

Nos últimos anos, a terapia de ressincronização cardíaca tem sido uma conduta bem definida para pacientes com insuficiência cardíaca (IC), classe funcional III ou IV, que não tiveram resposta à otimização terapêutica. Estudos já mostraram esta eficácia, mas um grupo bem definido de miocardiopatias dilatadas com áreas de fibrose não tem tido o mesmo sucesso (30 por cento-40 por cento). Por isto decidimos associar a estes pacientes o implante de células-tronco. A partir de 04/2005 iniciamos estes implantes em pacientes (pcs.) com IC, classe funcional III/IV, otmizados e fração de ejeção <35 por cento com dissincronia ventricular. Total de 25 pacientes (16 homens), idade entre 30-80anos (média de 58) e 44 por cento de etiologia chagásica (11 pcs). Ressincronização e terapia celular foram por via epicárdica (12 pcs) e endocárdica (13 pcs) e a seleção celular pela filtração por Ficoll e 3.0x10(8) de células infundidas CD34 + (5,0 x 10(6)) e CD 133 + (2,5 x 10(6)). Após 24 meses de evolução observamos que 84,8 por cento destes pcs permaneceram vivos e a mortalidade (15,2 por cento) não estava relacionada ao procedimento. A sobrevida dos portadores de marcapasso foi maior (52,4 por cento) e os chagásicos tiveram uma pior sobrevida, mas com teste de long-rank de 0,218 e 0,626, respectivamente. Nos casos de pcs com ressincronizador isoladamente, os resultados não parecem ser tão bons,e nós optamos pela associação de células-tronco como uma alternativa segura e adequada para estes graves pcs com insuficiência cardíaca e miocardiopatia dilatada.

In recent years cardiac resynchronization therapy has been a well defined approach for patients with functional class III or IV heart failure (HF) who do not respond to optimized therapy. Studies have shown efficacy, but one well defined group of dilated heart diseases with large areas of fibrosis has not had the same success rates (30-40 percent). Hence, we decided to associate stem cell transplant in these patients. Since April 2005 we started to transplant stem cells in 25 patients (16 men) with NYHA III/IV HF with ejection fraction < 35 percent and ventricle dyssynchrony. Their ages varied between 30 and 80, (mean age 58) and 44 percent (11 patients) had a chagasic etiology. Resynchronization and stem cell therapy were performed via epicardial (12) or endocardial (13) approaches. Cell selection was by Ficoll filtration with a mean number of 3.0 x 108 cells infused: CD 34+ (5.0 x 106) and CD 133+ (2.5 x 106). In 24 months of evolution we noticed that 84.8 percent of these patients survived and mortality (15.2 percent) was not related to the procedure. The survival of pacemaker recipients (52.4 percent) was higher than another similar group that did not receive pacemaker implants and chagasic patients survived less than the ischemic and the idiopathic patients (42.4 percent, 55.6 percent and 77.2 percent, respectively). In spite of these initial differences, there was no statistical difference between these groups (log-rank test of P=0.218 and P= 0.626, respectively). In cases that resynchronization was performed in isolation, the results do not seem to be satisfactory so we chose to associate stem cell therapy as a safe and adequate alternative for patients with HF and dilated myocardiopathy.