Effects of Calendula officinalis extract on liver histopathology, lipid profile, and oxidative stress in rats submitted to a diet rich in cholesterol and carbohydrates.

PURPOSE: To evaluate the modulatory properties of Calendula officinalis L. (Asteraceae) (C. officinalis) extract on cafeteria diet-fed rats. METHODS: A cafeteria diet was administered ad libitum for 45 days to induce dyslipidemia. Then, the rats were treated with the formulations containing C. officinalis in the doses of 50, 100, and 150 mg/kg or only with the vehicle formulation; the control group received a commercial ration. RESULTS: The cafeteria diet decreased glutathione S-transferase activity and high-density lipoprotein plasmatic levels and damaged the hepatic architecture. The C. officinalis extract was able to reduce lipid infiltration in liver tissue and to modulate oxidative stress and lipid profile markers. CONCLUSIONS: The correlations between the variables suggest a pathological connection between oxidative stress markers and serum lipid profile.

Brain-derived neurotrophic factor (BDNF) levels in children and adolescents before and after stimulant use a systematic review and metanalysis.

BACKGROUND: Attention deficit hyperactivity disorder (ADHD) is a neurodevelopmental disorder associated with cognitive, social, and academic impairment. Neurotrophins, particularly brain-derived neurotrophic factor (BDNF), have been implicated in the pathophysiology of ADHD and response to stimulant treatment. This review aims to investigate the relationship between BDNF levels in ADHD before and after treatment with stimulants in childhood. METHODS: This systematic review followed PRISMA-P guidelines and included 19 studies from PubMed, EMBASE, Cochrane, Capes Periodic, and Lilacs databases. The studies were evaluated for risk of bias and level of evidence. RESULTS: There was no significant difference in peripheral BDNF levels in ADHD children before or after methylphenidate treatment. Additionally, there was no statistically significant difference in BDNF levels between children with ADHD and controls. DISCUSSION: Understanding the role of BDNF in ADHD may provide insight into the disorder's pathophysiology and facilitate the development of biological markers for clinical use. CONCLUSION: Our findings suggest that BDNF levels are not significantly affected by methylphenidate treatment in ADHD children and do not differ from controls. SYSTEMATIC REVIEW REGISTRATION: "Brain-derived neurotrophic factor (BDNF) levels in children and adolescents before and after stimulant use: a systematic review". Number CRD42021261519.

Effects of Calendula officinalis extract on liver histopathology, lipid profile, and oxidative stress in rats submitted to a diet rich in cholesterol and carbohydrates

ABSTRACT Purpose: To evaluate the modulatory properties of Calendula officinalis L. (Asteraceae) (C. officinalis) extract on cafeteria diet-fed rats. Methods: A cafeteria diet was administered ad libitum for 45 days to induce dyslipidemia. Then, the rats were treated with the formulations containing C. officinalis in the doses of 50, 100, and 150 mg/kg or only with the vehicle formulation; the control group received a commercial ration. Results: The cafeteria diet decreased glutathione S-transferase activity and high-density lipoprotein plasmatic levels and damaged the hepatic architecture. The C. officinalis extract was able to reduce lipid infiltration in liver tissue and to modulate oxidative stress and lipid profile markers. Conclusions: The correlations between the variables suggest a pathological connection between oxidative stress markers and serum lipid profile.

Morphophysiological alterations in fruit-eating bats after oral exposure to deltamethrin.

Deltamethrin (DTM) is a synthetic pyrethroid widely used in the cultivation and management of several crops due to its insecticidal action. Application to crops of pyrethroids such as DTM can result in the exposure of water and fruit consumed by fruit bats having a high pyrethroid content which may be harmful. Therefore the objective of this study was to evaluate the effects of short-term oral exposure of the fruit-eating bats (Artibeus lituratus) to two concentrations of DTM (0.02 and 0.04 mg/kg of papaya) on histopathology of the intestine, liver and kidney. The intestine of the animals exposed to both concentrations showed inflammatory infiltrate, degeneration, necrosis and goblet cell hyperplasia as the most frequent pathologies. Besides, the acid mucins showed an increase in the frequency of non-viable cells. The liver showed hepatocyte vacuolizatio and nuclear enlargement, as well as inflammatory infiltrate and steatosis. The kidneys of the exposed animals showed and inflammatory infiltrate, benign nephrosclerosis, vacuolization and necrosis. Also, DTM reduced nitric oxide synthesis, decreased glomerular diameter and increased glycogen percentage in the proximal tubules. Our results suggest that acute exposure to DTM at low concentrations has the potential to induce pronounced histopathological changes in vital organs, such as intestine, liver and kidney of fruit-eating bats.

Protective immunity triggered by ectonucleoside triphosphate diphosphohydrolase-based biopharmaceuticals attenuates cardiac parasitism and prevents mortality in Trypanosoma cruzi infection.

Chagas disease is a potentially fatal infection in 21 endemic Latin America countries for which the effectiveness of reference antiparasitic chemotherapy is limited. Thus, we developed three biopharmaceuticals and evaluated the effectiveness of different immunization strategies (recombinant protein NTPDase-1 [rNTPDase-1], DNA plasmid encoding Trypanosoma cruzi NTPDase-1 [TcNTPDase-1] and DNA-NTPDase-1 prime/rNTPDase-1 boost [Prime-boost]) based on the surface ecto-nucleoside triphosphate diphosphohydrolase (ecto-NTPDase) enzyme of T. cruzi in animals challenged with a virulent strain (Y) of this parasite. BALB/c mice were immunized three times at 30 days intervals, challenged with T. cruzi 15 days after the last immunization, and euthanized 30 days after T. cruzi challenge. Our results showed limited polarization of specific anti-ecto-NTPDase immunoglobulins in mice receiving both immunization protocols. Conversely, the Prime-boost strategy stimulated the Th1 protective phenotype, upregulating TNF-α and downregulating IL-10 production while increasing the activation/distribution of CD3+/CD8+, CD4+/CD44hi and CD8+/CD44hi/CD62L cells in immunized and infected mice. Furthermore, IL-6 and IL10 levels were reduced, while the distribution of CD4+/CD44hi and CD3+/CD8+ cells was increased from rNTPDase-1 and DNA-NTPDase1-based immunization strategies. Animals receiving DNA-NTPDase1 and Prime-boost protocols before T. cruzi challenged exhibited an enhanced immunological response associated with IL-17 upregulation and remarkable downregulation of heart parasitism (T. cruzi DNA) and mortality. These findings indicated that NTPDase-1 with Prime-boost strategy induced a protective and sustained Th17 response, enhancing host resistance against T. cruzi. Thus, ecto-NTPDase is a potentially relevant and applicable in the development of biopharmaceuticals with greater immunoprophylactic potential for Chagas disease.

Is There a Connection between the Metabolism of Copper, Sulfur, and Molybdenum in Alzheimer's Disease? New Insights on Disease Etiology.

Alzheimer's disease (AD) and other forms of dementia was ranked 3rd in both the Americas and Europe in 2019 in a World Health Organization (WHO) publication listing the leading causes of death and disability worldwide. Copper (Cu) imbalance has been reported in AD and increasing evidence suggests metal imbalance, including molybdenum (Mo), as a potential link with AD occurrence.We conducted an extensive literature review of the last 60 years of research on AD and its relationship with Cu, sulfur (S), and Mo at out of range levels.Weanalyzed the interactions among metallic elements' metabolisms;Cu and Mo are biological antagonists, Mo is a sulfite oxidase and xanthine oxidase co-factor, and their low activities impair S metabolism and reduce uric acid, respectively. We found significant evidence in the literature of a new potential mechanism linking Cu imbalance to Mo and S abnormalities in AD etiology: under certain circumstances, the accumulation of Cu not bound to ceruloplasmin might affect the transport of Mo outside the blood vessels, causing a mild Mo deficiency that might lowerthe activity of Mo and S enzymes essential for neuronal activity. The current review provides an updated discussion of the plausible mechanisms combining Cu, S, and Mo alterations in AD.

Optimal Cutoff of the TG/HDL-c ratio for Cardiovascular Risk in Hypertensive and Diabetic Patients Monitored by Primary Health Care in a city in Minas Gerais

Abstract Background The analysis of the atherogenic potential of the lipid profile for biomarkers, such as the TG/HDL-c ratio, predicts cardiovascular risk better than isolated lipids. Objective To identify the TG/HDL-c cutoff points for multiple risks (hypertension, Diabetes Mellitus, obesity) and to evaluate the association between sociodemographic, clinical, laboratory, anthropometric, and life habit variables and the TG/HDL-c ratio in hypertensive and/or diabetic individuals in the context of Primary Health Care. Methods This was a cross-sectional study with 833 hypertensive and/or diabetic patients, conducted between August 2017 and April 2018. The cutoff point of the TG/HDL-c were obtained by the ROC curve. Cardiovascular risk was discriminated by TG/HDL-c, categorized by the cutoff and evaluated in relation to multiple risks. The magnitude of the association between TG/HDL-c and independent variables was estimated by logistic regression. The significance level of p<0.05 was adopted for all tests. Results The cutoff values of TG/HDL-c (3.26 for men and 2.72 for women) were more sensitive and less specific than those in the literature. Women (OR=1.90 and 95% CI 1.13-3.20) and men (OR=4.58 and 95% CI 1.78-11.76) with multiple risks, and white men, alcohol users, with a history of stroke, had a higher chance of altered GT/HDL-c. Increases in glycosylated hemoglobin, glycemia, and phosphorus in women, and cholesterol, glycemia, and microalbuminuria in men increased the chances of altered TG/HDL-c. Being a former smoker and black reduced the chance of altered TG/HDL-c in women. Conclusions TG/HDL-c proved to be a good indicator for habitual use in Primary Care.

Conhecimento geral, atitude psicológica e sua associação com a concentração de HbA1C em pacientes portadores de Diabetes Mellitus tipo 2 / General knowledge, psychological attitude and its association with the concentration of HbA1C in patients with Type 2 Diabetes Mellitus

Objetivo: Diabetes mellitus (DM) é uma doença relacionada a várias complicações e alta morbimortalidade, com comprometimento significativo da qualidade de vida do paciente, afetando sua saúde física e mental, bem como sua motivação ao autocuidado. Este estudo objetivou avaliar o conhecimento geral e a atitude psicológica para o DM, correlacionando a concentração de hemoglobina glicada (HbA1C) com escores do DKN-A e ATT-19 de pacientes portadores de DM tipo 2 (DM2). Métodos: Estudo observacional transversal com amostra de 65 participantes, 24,6% homens e 75,4% mulheres, com idade média de 57±12, avaliados entre novembro/2016 e fevereiro/2017. A avaliação do conhecimento geral e da atitude psicológica para o DM foi realizada pela aplicação do Diabetes Knowledge Questionnaire (DKN-A) e do Diabetes Attitude Questionnaire (ATT-19). O SPSS 20.0 foi utilizado para as análises estatísticas. Resultados: Para os escores de conhecimento (r=0,005; p=0,966) e de atitude (r=0,034; p=0,790) não houve correlação significante com a HbA1C, sendo que (94%) dos pacientes não atingiram o ponto de corte do ATT-19. Considerando o grupo com pontuação >8 para o escore de conhecimento, foi observada diferença negativa e significativa entre este escore e o de atitude (r=-0,444; p=0,0018). Conclusão: O conhecimento em relação à doença não traduziu em melhor controle da glicemia na população estudada, sugerindo que o controle eficiente da HbA1C não depende exclusivamente do conhecimento sobre a doença, mas sim de diversos outros fatores, que também precisam ser considerados na conduta do DM2.

Objective: Diabetes mellitus (DM) is a disease related to several complications and high morbidity and mortality, with significant impairment of the patient's quality of life, affecting his physical and mental health, as well as his motivation to self-care. The aim of this study was to evaluate the knowledge and attitude about type 2 DM (DM2) in its patients, correlating Diabetes Knowledge Questionnaire (DKN-A) and Diabetes Attitudes Questionnaire (ATT-19) scores with the concentration of glycated hemoglobin (HbA1c). Methods: Cross-sectional observational study with a sample of 65 participants, 24.6% male and 75.4% female, with a mean age of 57 ± 12, with low schooling, between November / 2016 and February / 2017. The evaluation of the knowledge and attitude about DM was carried out by the application of the DKN-A and ATT-19. SPSS 20.0 was used for statistical analysis. Results: There were no statistically significant correlations with HbA1c (r = 0.005, p = 0.966) and attitude (r = 0.034, p = 0.790), and 61 patients (94%) did not reach the cutoff point of ATT-19. Considering the group with scores higher than 8 points for the knowledge score, a negative and significant correlation was observed between this score and the attitude score (r = -0.444; p = 0.0018). Conclusion: Knowledge and attitude towards the disease did not translate into better glycemic control in the study population. The efficient control of HbA1c is multifactorial, involving genetic, comorbid, behavioral, drugs, health services, among others, that should be considered in the conduct.

Incidence and Prediction of Unstable Blood Glucose Level among Critically Ill Patients: A Cohort Study.

PURPOSE: To evaluate the incidence and the prediction of unstable blood glucose level among critically ill patients hospitalized in an intensive care unit. METHODS: A cohort study was conducted with 62 adult patients hospitalized at an intensive care unit of a hospital located in Minas Gerais, Brazil, between March and July of 2017. Patient's demographic information, along with scores for Simplified Acute Physiology Score III, primary medical diagnosis, discharge status, diagnosis of diabetes and/or sepsis, length of stay, glycemic variability, type of nutrition, types of medications and treatments, and oxygen therapy were collected daily. A daily venous blood sample was collected to measure blood glucose levels during the patient's hospitalization period. Bivariate analysis was used to explore the association among the potential diagnostic indicators and the outcome of unstable blood glucose levels. Multivariate Cox regression was used to identify the potential predictors for the outcome. FINDINGS: Of the total of 62 participants, 45.1% (n=28) had unstable blood glucose level. Among the 28 patients with unstable blood glucose levels, half of them (n=14, 50%) had hypoglycemia and the other half had hyperglycemia (n=14, 50%). Decreased number of days hospitalized and the use of intensive glucose control with regular insulin were associated with decreased odds of developing hyperglycemia. The presence of mechanical ventilation was associated with a higher risk for the development of hypoglycemia. CONCLUSIONS: This study provides knowledge and evidence of diagnostic indicators for unstable blood glucose levels that are not currently included in the NANDA-International terminology for the nursing diagnosis Risk for unstable blood glucose level (00179). IMPLICATIONS FOR NURSING PRACTICE: This study identified important diagnostic indicators that nurses can observe during the assessment to identify patients that are at risk for developing unstable blood glucose level and provide the appropriate care.

OBJETIVO: Avaliar a incidência e predição de glicemia instável em pacientes adultos internados em uma Unidade de Terapia Intensiva. MÉTODO: Estudo de coorte concorrente, realizada entre março e julho de 2017, com amostra de 62 pacientes. As variáveis independentes coletadas da admissão do paciente a unidade foram sexo, raça, idade, Simplified Acute Physiology Score III, diagnóstico de internação, tipo de saída, diabetes, sepse, tempo de permanência, variabilidade glicêmica, tipo de nutrição, medicamentos e terapêutica, e oxigenoterapia (cateter nasal e ventilação mecânica). Os dados foram analisados por estatística descritiva e análise de sobrevida com teste de Log-rank e regressão multivariada de Cox. RESULTADOS: Dos 62 pacientes, 45,1% (n=28) desenvolveram a glicemia instável. Dos 28 pacientes com glicemia instável, metade (n=14, 50%) apresentou hipoglicemia, e a outra metade apresentou hiperglicemia (n=14, 50%). Menor tempo de permanência e controle estrito da glicemia foram fatores de proteção para desenvolvimento de hiperglicemia. Ventilação mecânica foi associada ao desenvolvimento de hipoglicemia. CONCLUSÃO: Este estudo fornece conhecimento e evidência sobre indicadores diagnósticos para o desenvolvimento de glicemia instável, que atualmente não estão incluídos na terminologia da NANDA-Internacional para o diagnóstico de enfermagem Risco de glicemia instável. IMPLICAÇÕES PARA ENFERMAGEM: Este trabalho contribui para o aperfeiçoamento da Taxonomia da NANDA-Internacional e, por conseguinte, da linguagem padronizada de enfermagem.

Neuromuscular activation analysis of the trunk muscles during hippotherapy sessions.

INTRODUCTION: Hippotherapy allows the development of affective, sensory, motor, and cognitive areas, besides providing the practitioner with several movements and stimuli necessary for therapeutic progress. However, there is a limited amount of scientific evidence regarding the suitability of the mount material, mount type, and hippotherapy session duration, as well as regarding the activation of specific muscle groups during the practice and its applicability to activities of daily living. OBJECTIVE: This study aimed to study the neuromuscular activation behavior of the iliocostalis, longissimus, multifidus, and upper trapezius muscles of children during four hippotherapy session time points using a functional task. It also compared two different mount materials for riding. METHODOLOGY: A total of 30 children were randomly assigned to one of three groups: Saddle Hippotherapy Group, Blanket Hippotherapy Group, and Control Group. Data were collected with an electromyograph in a functional task that comprised trunk movements to pick up an object. Assessments took place at four times during the session. RESULTS: There was a significant increase in the neuromuscular activation of the iliocostalis, longissimus, and multifidus muscles after a 30-min session. The trapezius muscle showed increased neuromuscular activation after only 10 min. It continued to increase (but without a statistical difference) after and 20 and 30 min. CONCLUSION: Hippotherapy promoted neuromuscular activation of the trunk muscles in children, assessed through a functional task, and was influenced by both session time and mount material. Specifically, the greater neuromuscular performance occurred when an exercise was performed using saddle and stirrup and lasted 30 min.

Oxidative stress markers in cognitively intact patients with diabetic neuropathy.

Various forms of vascular injury are frequently associated with type-2 diabetes mellitus (DM2). Macro-angiopathy has alarming signs and symptoms such as those seen with stroke or heart attack, however the presentation of small vessel disease is generally more subtle and therefore usually unnoticed for a long period of time. While it may affect any organ, complications involving the nervous system such as diabetic poly-neuropathy (DPN) are especially debilitating, and it may also be a risk factor for other brain disorders such as dementia. The underlying mechanisms are likely to be multi-faceted, but piling evidence indicates oxidative stress as one of the crucial factors. Here we evaluate the oxidative profile of patients with DM2. The total anti-oxidant capacity appears to be reduced in DM2 with or without complications. Of the specific bio-markers studied, the levels of tissue-damage indicator malon-dialdehyde (MDA) were significantly lower in the DM2 + DPN population only. These results suggest that diabetic patients present with wavering oxidative status, and the low MDA concentrations in patients with complications such as DPN may represent either an exhausted anti-oxidative defense system or a response to anti-inflammatory medications. The findings may also support the use of anti-oxidants such as vitamins A and E.

Prevalence of vitamin D deficiency and its relationship with factors associated with recurrent wheezing.

OBJECTIVE: To determine the prevalence of vitamin D deficiency/insufficiency in children 0-18 years of age with recurrent wheezing and/or asthma residing in the microregion of Viçosa, Minas Gerais, Brazil, and treated at a referral center, and to determine its association with major risk factors for wheezing. METHODS: A cross-sectional study was performed using a semi-structured questionnaire, which was administered by trained interviewers to the legal guardians of the study participants. Data were obtained regarding general characteristics of recurrent wheezing; general sociodemographic, environmental, and biologic factors; and atopy-related factors. The magnitude of the statistical association was assessed by calculating ORs and their corresponding 95% CIs by using multiple logistic regression. RESULTS: We included 124 children in the study. The prevalence of vitamin D deficiency/insufficiency in the sample was 57.3%. Vitamin D deficiency/insufficiency was found to be associated with wheezing in the first year of life, personal history of atopic dermatitis, environmental pollution, and vitamin D supplementation until 2 years of age. CONCLUSIONS: The prevalence of vitamin D deficiency/insufficiency was high in our sample. Vitamin D concentrations were directly associated with vitamin D supplementation until 2 years of age and were inversely associated with wheezing events in the first year of life, personal history of atopic dermatitis, and environmental pollution.

Prevalence of vitamin D deficiency and its relationship with factors associated with recurrent wheezing / Prevalência de deficiência de vitamina D e sua relação com fatores associados à sibilância recorrente

ABSTRACT Objective: To determine the prevalence of vitamin D deficiency/insufficiency in children 0-18 years of age with recurrent wheezing and/or asthma residing in the microregion of Viçosa, Minas Gerais, Brazil, and treated at a referral center, and to determine its association with major risk factors for wheezing. Methods: A cross-sectional study was performed using a semi-structured questionnaire, which was administered by trained interviewers to the legal guardians of the study participants. Data were obtained regarding general characteristics of recurrent wheezing; general sociodemographic, environmental, and biologic factors; and atopy-related factors. The magnitude of the statistical association was assessed by calculating ORs and their corresponding 95% CIs by using multiple logistic regression. Results: We included 124 children in the study. The prevalence of vitamin D deficiency/insufficiency in the sample was 57.3%. Vitamin D deficiency/insufficiency was found to be associated with wheezing in the first year of life, personal history of atopic dermatitis, environmental pollution, and vitamin D supplementation until 2 years of age. Conclusions: The prevalence of vitamin D deficiency/insufficiency was high in our sample. Vitamin D concentrations were directly associated with vitamin D supplementation until 2 years of age and were inversely associated with wheezing events in the first year of life, personal history of atopic dermatitis, and environmental pollution.

RESUMO Objetivo: Determinar a prevalência da deficiência/insuficiência de vitamina D em indivíduos com sibilância recorrente e/ou asma com idade de 0-18 anos e residentes na microrregião de Viçosa (MG) atendidos em um centro de referência e determinar sua associação com os principais fatores de risco para sibilância. Métodos: Estudo transversal utilizando um questionário semiestruturado por entrevistadores treinados, aplicado aos responsáveis pelos participantes do estudo. Foram obtidas informações sobre características gerais da sibilância recorrente, fatores sociodemográficos, ambientais e biológicos gerais e aqueles relacionados à atopia. A magnitude da associação estatística foi avaliada por meio do cálculo da OR e IC95% obtidos por regressão logística múltipla. Resultados: Foram incluídos 124 indivíduos no estudo. A prevalência da deficiência/insuficiência de vitamina D na amostra foi de 57,3%. Observaram-se associações da deficiência/insuficiência de vitamina D com sibilância no primeiro ano de vida, antecedentes pessoais de dermatite atópica, poluição ambiental e suplementação de vitamina D até os 2 anos de idade. Conclusões: A prevalência de deficiência/insuficiência de vitamina D foi alta em nossa amostra. As concentrações de vitamina D foram associadas diretamente com a suplementação de vitamina D até os 2 anos de idade e inversamente com eventos de sibilância no primeiro ano de vida, antecedentes pessoais de dermatite atópica e poluição ambiental.

Análise da patogênese da doença de Alzheimer: revisão narrativa da literatura / Analysis of Alzheimer's disease pathogenesis: narrative literature review

Introdução: A patogênese de doenças neurodegenerativas, como a doença de Alzheimer (DA), têm sido cada vez mais objeto de estudo em inúmeras pesquisas científicas. Objetivo: A presente revisão narrativa teve por objetivo descrever a contribuição de fatores como o estresse oxidativo e o processo de netose para a patogênese de doenças neurodegenerativas, como a DA. Material e Métodos: Realizou-se revisão de literatura de forma não sistemática nos bancos de dados das plataformas PubMed e Scielo com os seguintes descritores: Alzheimer + oxidative stress, Alzheimer + netosis e Alzheimer + inflammation, em periódicos nacionais e internacionais de 2000 a 2018. Ao final da seleção foram utilizados 26 artigos nesta revisão. Resultados: Diversos estudos demostraram importantes componentes ligados à neuroinflamação como peças-chave para o entendimento do acometimento neurológico causado pela DA, tais como, o estresse oxidativo, respostas inflamatórias por parte de neutrófilos e da micróglia, alterações vasculares e da barreira hematoencefálica. Conclusão: Em função de tais descobertas, faz-se necessário a continuidade e o avanço no entendimento da fisiopatologia da DA bem como da busca por eficazes abordagens terapêuticas para esta doença.

Introduction: The pathogenesis of neurodegenerative diseases, such as Alzheimer's disease (AD), has been increasingly studied in numerous scientific studies. Objective: The purpose of this review was to describe the contribution of factors such as oxidative stress and the netosis process for the pathogenesis of neurodegenerative diseases such as AD. Material and Methods: A non-systematic bibliographic review was carried out in the databases PubMed and Scielo with the following descriptors: Alzheimer + oxidative stress, Alzheimer + netosis and Alzheimer + inflammation, in national and international scientific journals from 2000 to 2018. At the end of the selection 26 articles were used in this review. Results: Several studies have demonstrated important components linked to neuroinflammation as key components for the understanding of neurological involvement caused by AD, such as oxidative stress, neutrophil and microglial inflammatory responses, vascular and blood-brain barrier changes. Conclusion: Due to these findings, it is necessary to continue and advance the understanding of the pathophysiology of AD as well as the search for effective therapeutic approaches for this disease.

Bacteriophage Isolated from Sewage Eliminates and Prevents the Establishment of Escherichia Coli Biofilm.

Purpose: Biofilm growth exerts a negative impact on industry and health, necessitating the development of strategies to control. The objective of this work was study the lytic activity of the phage isolated from the sewage network in the formation and degradation of Escherichia coli biofilms. Methods: E. coli cultures were incubated in 96-well polystyrene microplates under controlled conditions to evaluate the biofilm formation. The E. coli cultures and established biofilms were treated with the suspensions of the vB_EcoM-UFV017 (EcoM017) bacteriophage obtained from sewage for 24 hours. The E. coli bacterial density was measured using absorbance at 600 nm and the biofilms were measured by crystal violet staining. Polystyrene coupons were used as support for Scanning Electron Microscopy and Confocal Microscopy to evaluate biofilm formation. Results: The E. coli strains formed biofilms in polystyrene microplates after 48 hours' incubation. The highest EcoM017 phage titer, in the prevention and degradation experiments, reduced the bacterial growth and the quantity of biofilm formed by E. coli in 90.0% and 87.5%, respectively. The minimum dose capable of reducing the biofilms of this bacterium was 101 PFU/mL after 24 hours. The preformed E. coli biofilm mass was reduced 79% post exposure to the phage in the degradation assay. Microscopic analysis confirmed the results obtained in the plates assays. Conclusion: The EcoM017 phage prevented biofilm formation and degraded the E. coli-established ones. The EcoM017 phage isolated from sewage can reduce bacterial attachment and lyse the E. coli associated biofilm cells, offering biotechnological potential applicability for this phage.

Exposure to deltamethrin induces oxidative stress and decreases of energy reserve in tissues of the Neotropical fruit-eating bat Artibeus lituratus.

Deltamethrin (DTM) is a synthetic pyrethroid insecticide highly used by farmers and home users. This pesticide has lipophilic properties that facilitate a high absorption and can cause toxicity in non-target organisms. During foraging, the fruit-eating bats Artibeus lituratus are exposed to pesticides. However, the knowledge of the toxicity of pesticides on the physiology of bats is relatively scarce. This study aimed to check the toxicity of short-term exposure to low concentration of DTM on fruit-eating bat A. lituratus. After seven days of exposure to two doses of DTM (0.02 and 0.04mg/kg of papaya), the fruit bats showed an increase in the enzyme aspartate aminotransferase, alanine aminotransferase, and hyperglycemia. The liver and pectoral muscle presented oxidative stress. In the liver, the hydrogen peroxide (H2O2) and nitric oxide (NO) were increased as well as the antioxidant glutathione (GSH), the activity of glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase (CAT) but in a lesser extent. Yet, total lipids were increased while hepatic glycogen content is reduced. The pectoral muscle showed NO, SOD, CAT, malondialdehyde (MDA), and carbonyl increased protein levels in both concentrations of DTM. All these results show that low doses of DTM can cause hepatic and muscular toxicity and induce changes in carbohydrate metabolism. Physiological changes caused by exposure to DTM in bats may have direct consequences in flight capacity, reproduction, and metabolism of these animals.

Prevalência de conjuntivite adenoviral em clínica oftalmológica no município de Viçosa (MG) / Prevalence of adenoviral conjunctivitis at the ophthalmologic clinic on municipality of Viçosa/MG

Objetivo: Avaliar a prevalência de Adenovírus como agente etiológico da conjuntivite, em clínica médica oftalmológica especializada, em Viçosa, Minas Gerais, Brasil. Métodos: Amostras da secreção conjuntival de 91 pacientes clinicamente diagnosticados com conjuntivite foram submetidos à reação em cadeia da polimerase (PCR), utilizando primers degenerados para a região codificadora do gene da proteína estrutural II. Posteriormente as amostras positivas foram submetidas a sequenciamento e genotipagem. Resultados: A análise dos resultados de PCR revelou prevalência de 36,3% de Adenovírus. Não havendo distinção entre os sexos e com maior prevalência na faixa etária de 26 a 65 anos com 60,60% dos casos positivos. O sequenciamento dos casos positivos por Adenovírus revelaram a presença dos sorotipos 3, 4, 7, 8 e 34 circulante na região. Conclusão: No município de Viçosa, dois em cada cinco casos de conjuntivite são de etiologia adenoviral.

Objective: The aim of this study was to evaluate the prevalence of Adenovirus as a etiologic agent of conjunctivitis on a ophthalmic clinic in Viçosa, Minas Gerais, Brazil. Methods: Samples of conjunctival secretion from 91 patients clinically diagnosed with conjunctivitis were subjected to polymerase chain reaction (PCR) using degenerate primers targeted to the gene encoding the structural protein II. Positive samples were subsequently subjected to sequencing and genotyping. Results: PCR results showed 36.3% prevalence of Adenovirus. No differences between the sexes and was found to be higher in the age group 26-65 years with 60.60% of the positive cases. Sequencing of positive cases showed the presence of Adenovirus serotypes 3, 4, 7, 8, and 34 circulating in the region. Conclusion: In Viçosa two in five cases of conjunctivitis has Adenovirus as etiologic agent.

Construction of recombinant Kluyveromyces marxianus UFV-3 to express dengue virus type 1 nonstructural protein 1 (NS1).

The yeast Kluyveromyces marxianus is a convenient host for industrial synthesis of biomolecules. However, despite its potential, there are few studies reporting the expression of heterologous proteins using this yeast. Here, we report expression of a dengue virus protein in K. marxianus for the first time. The dengue virus type 1 nonstructural protein 1 (NS1) was integrated into the K. marxianus UFV-3 genome at the LAC4 locus using an adapted integrative vector designed for high-level expression of recombinant protein in Kluyveromyces lactis. The NS1 gene sequence was codon-optimized to increase the level of protein expression in yeast. The synthetic gene was cloned in frame with K. lactis α-mating factor signal peptide, and the recombinant plasmid obtained was used to transform K. marxianus UFV-3 by electroporation. The transformed cells, selected in yeast extract peptone dextrose containing 200 µg mL(-1) Geneticin, were mitotically stable. Analysis of recombinant strains by RT-PCR and protein detection using blot analysis confirmed both transcription and expression of extracellular NS1 polypeptide. After induction with galactose, the NS1 protein was analyzed by sodium dodecyl sulfate-PAGE and immunogenic detection. Protein production was investigated under two conditions: with galactose and biotin pulses at 24-h intervals during 96 h of induction and without galactose and biotin supplementation. Protease activity was not detected in post-growth medium. Our results indicate that recombinant K. marxianus is a good host for the production of dengue virus NS1 protein, which has potential for diagnostic applications.

Leishmania chagasi heparin-binding protein: Cell localization and participation in L. chagasi infection.

Visceral leishmaniasis is a fatal human disease caused by the intracellular protozoan parasite Leishmania chagasi that is captured by host cells in a process involving classics receptors mediated phagocytosis. The search for molecules involved in this process is important to design strategies to disease control. In this work, we verified the presence of heparin-binding protein (HBP) in L. chagasi promastigotes forms. HBP is a lectin of the group of ubiquitous proteins, whose main characteristic is to bind to carbohydrates present in glycoproteins or glycolipids, which is poorly studied in Leishmania species. L. chagasi HBP (HBPLc) was purified by affinity chromatography using heparin-agarose column in FPLC automated system. Its localization in the parasite was assessed by immunolabeling and electronic transmission microscopy tests using anti-HBPLc polyclonal antibodies, which showed HBP spread over the parasite outer surface and internally next to the kynetoplast. In addition, we verified that HBPLc participates in the process of parasite infection, since its blocking with heparin generated a partial reduction in the internalization of Leishmania by RAW macrophages "in vitro". According to these results, it is believed that, in further "in vivo" studies, interference on this parasitic protein may provide us prophylactic and therapeutic alternatives against visceral leishmaniasis.

Nasal vaccination with attenuated Salmonella expressing VapA: TLR2 activation is not essential for protection against R. equi infection.

Virulent strains of Rhodococcus equi have a large plasmid of 80-90kb, which encodes several virulence-associated proteins (Vap), including VapA, a lipoprotein highly associated with disease. We have previously demonstrated that oral immunisation with attenuated Salmonella enterica Typhimurium strain expressing the antigen VapA (STM VapA+) induces specific and long-term humoral and cellular immunity against R. equi. It was shown that VapA activates Toll-like receptor 2 (TLR2) on macrophages by establishing an interaction that ultimately favours immunity against R. equi infection. The purpose of this study was to evaluate the immune response triggered by nasal immunisation with STM VapA+ and to determine whether TLR2 supports the vaccine effect. We developed an optimised protocol for a single nasal immunisation that conferred protection against R. equi infection in mice, which was manifested by efficient R. equi clearance in challenged animals. Nasal vaccination with STM VapA+ has also induced protection in Tlr2(-/-) mice and mice with non-functional TLR4. Moreover, spleen cells of vaccinated mice augmented T-bet expression, as well as the production of IL-12, IFN-γ, nitric oxide and hydrogen peroxide. Notably, the population of CD4(+) T cells with memory phenotype significantly increased in the spleens of vaccinated mice challenged 1 or 5 months after immunisation. In these animals, the spleen bacterial burden was also reduced. When similar experimental procedures were performed in TLR2 knockout mice, an increase in CD4(+) T cells with memory phenotype was not observed. Consequently, we conclude that nasal vaccination with attenuated Salmonella expressing the R. equi virulence factor VapA confers long-lasting protection against experimental rhodoccocosis and that TLR2 engagement was not crucial to induce this protection but may be required for a long-term immune response.