Psychotropic drug use and suggestive depression symptoms associated with NR3C1 DNA methylation / Uso de drogas psicotrópicas e sintomas sugestivos de depressão associados à metilação do DNA do NR3C1

Introduction: Psychiatric disorders have become a global problem that leads millions of people to use psychotropic medications, especially benzodiazepines. The effects of these substances are widely known regarding tolerance and chemical dependence, however, from epigenetics perspective, there are still little known.Objective: To evaluate the association between psychotropic drug use, NR3C1 gene methylation and its relation with symptoms suggestive of depression in adult individuals assisted in the public health system.Methods: 385 adult volunteers (20-59 years) users of the Brazilian Unified Health System were recruited to evaluate socioeconomic, health, lifestyle conditions in a cross sectional study. BDI-II evaluated symptoms suggestive of depression and pyrosequencing evaluated NR3C1 DNA methylation. Bivariate and multivariate Poisson regression model with robust variance (p < 0.05) evaluated the association between psychotropic drug use and NR3C1 gene methylation.Results: Specific depressive symptoms such as irritability, insomnia and fatigability were associated with psychotropic drug use. Symptoms of past failure, indecision and loss of appetite were associated with hypermethylation patterns in CpGs 40 to 47 of NR3C1 gene. Moreover, psychotropic drug use is associated with 50% reduction in NR3C1 gene methylation, through model adjusted with socioeconomic, health and lifestyle confounding variables.Conclusions: Psychotropic drug use and depressive symptoms was associated with changes in NR3C1 DNA methylation. In this context, epigenetic modification resulting from psychotropic drug use and depressive symptoms could be considered, mainly in population studies with epigenetic evaluation, where these factors may be influencing the findings of future studies.

Introdução: os distúrbios psiquiátricos tornaram-se um problema global que leva milhões de pessoas ao uso de medicamentos psicotrópicos. Os efeitos dessas substâncias são amplamente conhecidos quanto à tolerância e dependência química, porém, do ponto de vista epigenético, ainda são pouco conhecidos.Objetivos: avaliar a associação entre o uso de drogas psicotrópicas, metilação do gene NR3C1 e sua relação com sintomas sugestivos de depressão em indivíduos entre 20 a 59 anos usuários da rede pública de saúde.Método: 385 voluntários de 20-59 anos, usuários do Sistema Único de Saúde brasileiro foram recrutados para avaliação das condições socioeconômicas, de saúde e de estilo de vida em estudo transversal. O BDI-II avaliou sintomas sugestivos de depressão e o pirosequenciamento avaliou a metilação do DNA de NR3C1. Modelo de regressão de Poisson bivariado e multivariado com variância robusta (p < 0,05) avaliou a associação entre o uso de drogas psicotrópicas e metilação do gene NR3C1.Resultados: sintomas depressivos específicos como irritabilidade, insônia e fadiga foram associados ao uso de medicamentos psicotrópicos. Sintomas de fracasso passado, indecisão e perda de apetite foram associados a padrões de hipermetilação nos CpGs 40 a 47 do gene NR3C1. Além disso, o uso de psicofármacos está associado à redução de 50% na metilação do gene NR3C1, por meio de modelo ajustado com variáveis de confusão socioeconômicas, de saúde e estilo de vida.Conclusão: o uso de drogas psicotrópicas e sintomas específicos depressivos foram associados a alterações na metilação do DNA de NR3C1.

Electrocardiographic changes and changes in cardiac lactate and troponin I levels associated with search and rescue physical activity in military dogs / Alterações eletrocardiográficas e nos níveis de lactato e troponina I cardíaca associadas à atividade física de busca e salvamento em cães militares

Rescue and recovery dogs intercalate the activity intensity developed, which also triggers significant metabolic changes in cardiac physiology. Thus, we evaluated the changes that search simulation causes in glucose, lactate, and cardiac troponin I level (cTnI) and the electrocardiographic and heart rate during the activity and recovery phase to predict the physiological adaptation to the exercise. Five healthy adult dogs from the Rescue and Recovery Service of Military Firefighters Corps were submitted to 60 minutes search operation simulation in the woods. They covered a forest area of approximately 50,000 m2. The dogs were loose and accompanied by their driver, and they could perform any physical activity. Were evaluated serum biochemical analysis of glucose, lactate, cardiac troponin I, electrocardiographic, and heart rate (rest, exercise phase, and recovery time). No changes in glucose levels, heart rate, and cardiac rhythm were detected. In comparison to baseline values, there is an increase: in lactate at the end of the exercise phase [EXER] (60'EXER), and in the recovery phase [RCT] at 30'RCT and 60'RCT, and cTnI at 60'RCT, 120'RCT, and 4hRCT. P wave duration was significantly higher at 60'EXER, 15'RCT, and 30'RCT, with no alterations in wave amplitude. QRS interval duration significantly increased at 30'RCT, and the ST segment presented a significant difference at 60'EXER, 15'RCT, and 60'RCT compared to the rest moment. The moderate alterations in lactate and cTnI and few alterations in the electrocardiographic and heart rate maintenance suggest the adaptation of rescue and recovery dogs to the type, intensity, and duration of search operation simulation performed.(AU)

Cães de busca e resgate intercalam a intensidade da atividade desenvolvida que desencadeia alterações metabólicas significativas, bem como na fisiologia cardíaca. Assim, foram avaliadas as alterações que a simulação de busca produz nos níveis de glicose, lactato, troponina I cardíaca (cTnI), bem como na frequência cardíaca e atividade eletrocardiográfica durante a fase de atividade e recuperação, a fim de predizer a adaptação fisiológica ao exercício. Cinco cães adultos saudáveis do Serviço de Resgate e Salvamento do Corpo de Bombeiros Militares foram submetidos à simulação de operação de busca de 60 minutos na mata e cobriram uma área florestal de aproximadamente 50.000 m2. Os cães estavam soltos, acompanhados pelo condutor e estavam livres para realizar qualquer tipo de atividade física. Foram avaliados os níveis séricos de glicose, lactato e troponina I cardíaca, atividade eletrocardiográfica e frequência cardíaca em repouso, na fase de exercício e no tempo de recuperação. Não foram detectadas alterações nos níveis de glicose, frequência cardíaca e ritmo cardíaco. Em comparação com os valores basais houve aumento de lactato ao final da fase de exercício [EXER] (60'EXER) e na fase de recuperação [RCT] aos 30'RCT e 60'RCT; e cTnI aos 60'RCT, 120'RCT e 4hRCT. Duração da onda P foi significativamente maior em 60'EXER, 15'RCT e 30'RCT, sem alterações na amplitude da onda. Duração do intervalo QRS teve aumento significativo em 30'RCT e o segmento ST apresentou diferença significativa em 60'EXER, 15'RCT e 60'RCT quando comparado ao basal. As alterações moderadas nos níveis de lactato e cTnI, bem como a pouca alteração na atividade eletrocardiográfica e manutenção da frequência cardíaca sugerem boa adaptação dos cães de busca e resgate ao tipo, intensidade e duração da operação de busca simulada realizada.(AU)

PAI-1 expression in intratumoral inflammatory infiltrate contributes to lymph node metastasis in oral cancer: A cross-sectional study.

INTRODUCTION: Immune cells contribute with mediators in the protein expression profile of the tumor microenvironment. Levels of plasminogen activator inhibitor-1 (PAI-1) are elevated in non-malignant inflammatory conditions; however, the association between PAI-1 expression and inflammation remains uncertain in oral squamous cell carcinoma (OSCC). This study aimed to investigate PAI-1 expression in mononuclear inflammatory cell infiltrate in OSCC and its role as a prognostic marker. METHODS: Samples were collected from patients with OSCC, treated surgically, and followed for 24 months after the procedure. Thirty-nine tumoral tissue were analyzed using immunohistochemistry. Correlation between protein expression, clinicopathological parameters, and the prognosis was investigated. RESULTS: Positive PAI-1 expression in mononuclear inflammatory cell infiltrate was significantly associated with lymph node status (p = 0.009) and with the cytoplasmic expression of vascular endothelial growth factor A (VEGFA) (p = 0.028). Multivariate analysis revealed weak PAI-1 expression as an independent marker for lymph node metastases, with approximately 8-fold increased risk compared to strong expression (OR = 8.60; CI = 1.54-48.08; p = 0.014). CONCLUSION: Our results suggest that the strong PAI-1 expression in intratumoral inflammatory infiltrate is an indicator of a better prognosis for patients diagnosed with oral squamous cell carcinoma.

Alcohol consumption, depression, overweight and cortisol levels as determining factors for NR3C1 gene methylation.

The NR3C1 glucocorticoid receptor (GR) gene is a component of the stress response system, which can be regulated by epigenetic mechanisms. NR3C1 methylation has been associated with trauma and mental issues, including depression, post-traumatic stress, anxiety, and personality disorders. Previous studies have reported that stressful events are involved in NR3C1 gene methylation, suggesting that its regulation under environmental effects is complex. The present study aimed to analyze associations involving stressors such as socioeconomic status, health conditions, and lifestyle in relation to NR3C1 methylation in adults. This study included 386 individual users of the Brazilian Public Unified Health System (SUS), and evaluated socioeconomic and health conditions, body mass index, cortisol levels, and lifestyle. Data were correlated with NR3C1 methylation, determined using DNA pyrosequencing. The results showed that alcohol consumption, overweight, and high cortisol levels were related to NR3C1 demethylation, while depression was related to its methylation. Habits, lifestyle, and health status may influence NR3C1 gene regulation via methylation, revealing the complexity of environmental impacts on NR3C1 methylation.

Elemental characterization of oral cavity squamous cell carcinoma and its relationship with smoking, prognosis and survival.

Oral cancer squamous cell carcinoma (OCSCC) mainly affects individuals aged between 50 and 70 years who consume tobacco and alcohol. Tobacco smoke contains hundreds of known toxic and carcinogenic molecules, and a few studies have sought to verify the relationship of such trace elements as risk or prognostic factors for head and neck cancer. We obtained 78 samples of tumor tissues from patients with OCSCC, and performed a qualitative elemental characterization using the micro X-Ray Fluorescence technique based on synchrotron radiation. We found the presence of magnesium, phosphorus, sulfur, chlorine, potassium, calcium, chromium, manganese, iron, zinc, cobalt, nickel, copper, arsenic and bromine in OCSCC samples. Magnesium, chlorine, chromium, manganese, nickel, arsenic and bromine are associated with smoking. We observed a significant association between relapse and chlorine and chromium. The presence of chlorine in the samples was an independent protective factor against relapse (OR = 0.105, CI = 0.01-0.63) and for best disease-free survival (HR = 0.194, CI = 0.04-0.87). Reporting for the first time in oral cancer, these results suggest a key relationship between smoking and the presence of certain elements. In addition, chlorine proved to be important in the context of patient prognosis and survival.

Morphometric and vascular analyses and MMP-2 expression in bladders of animals with bovine enzootic haematuria

Background: Bovine enzootic haematuria is a chronic, non-infectious disease caused by the ingestion of bracken Pteridiumaquilinum that contain ptaquiloside, substance responsible for the development of neoplasms in combination with bovinepapillomavirus infection. Studies examining metalloproteinases have shown that these enzymes may be useful as prognosticmarkers and many studies have correlated the intratumoural microvessel density and incidence of metastasis in a varietyof cancers, including bladder cancer. This study aimed to evaluate the urothelial morphology and vasculature and quantifythe expression of the enzyme metalloproteinase-2 in the bladder lesions of bovine with enzootic haematuria.Materials, Methods & Results: Forty bladders with macroscopic lesions were subjected to routine histological processing. The material was stained using the Hematoxylin-Eosin (HE) and Weigerts resorcin-fuchsin staining methods. Themorphometric analysis included the measurement of the total thickness of the urothelium. Microscopically, the lesionswere differentiated as neoplastic or non-neoplastic, and the vascular changes were characterised as vascular proliferation,ectasia, dilation and thickening and the material stained using the Weigerts resorcin-fuchsin method was used to evaluatesclerosis and the vascular fibroelastosis. The metalloproteinase expression was evaluated using an anti-metalloproteinase-2antibody. The main non-neoplastic lesions found in the urothelium included clear cell metaplasia, dysplasia, hyperplasia,haemorrhage, cystitis cystica and Brunns nests and the neoplastic lesions were haemangioma, myxoma, transitional cellcarcinoma, adenocarcinoma, in situ carcinoma and haemangiosarcoma. The urothelium measurements revealed differencesbetween the epithelial thicknesses of bladders with and without neoplasms. The bladder vasculature evaluation revealed...(AU)

Morphometric and vascular analyses and MMP-2 expression in bladders of animals with bovine enzootic haematuria

Background: Bovine enzootic haematuria is a chronic, non-infectious disease caused by the ingestion of bracken Pteridiumaquilinum that contain ptaquiloside, substance responsible for the development of neoplasms in combination with bovinepapillomavirus infection. Studies examining metalloproteinases have shown that these enzymes may be useful as prognosticmarkers and many studies have correlated the intratumoural microvessel density and incidence of metastasis in a varietyof cancers, including bladder cancer. This study aimed to evaluate the urothelial morphology and vasculature and quantifythe expression of the enzyme metalloproteinase-2 in the bladder lesions of bovine with enzootic haematuria.Materials, Methods & Results: Forty bladders with macroscopic lesions were subjected to routine histological processing. The material was stained using the Hematoxylin-Eosin (HE) and Weigert’s resorcin-fuchsin staining methods. Themorphometric analysis included the measurement of the total thickness of the urothelium. Microscopically, the lesionswere differentiated as neoplastic or non-neoplastic, and the vascular changes were characterised as vascular proliferation,ectasia, dilation and thickening and the material stained using the Weigert’s resorcin-fuchsin method was used to evaluatesclerosis and the vascular fibroelastosis. The metalloproteinase expression was evaluated using an anti-metalloproteinase-2antibody. The main non-neoplastic lesions found in the urothelium included clear cell metaplasia, dysplasia, hyperplasia,haemorrhage, cystitis cystica and Brunn’s nests and the neoplastic lesions were haemangioma, myxoma, transitional cellcarcinoma, adenocarcinoma, in situ carcinoma and haemangiosarcoma. The urothelium measurements revealed differencesbetween the epithelial thicknesses of bladders with and without neoplasms. The bladder vasculature evaluation revealed...

Biometry of adrenal glands of mixed-breed dogs

Background: Differences of the size of the adrenal glands, in healthy dogs or in endocrinopathies, undermine correct diagnosis of endocrine disorders and evaluations of the adrenal cortex in relation to its size and possible correlation with endocrinopathies are rare. The aim of the present study was to perform measurements of the length, width, thickness and weight of the adrenal glands of young, adult and elderly mixed-breed dogs and correlate them with the age, sex and weight of animals. In addition, the areas occupied by the zona glomerulosa, zona fasciculata, and zona reticularis of the adrenal cortex were measured in order to establish a microscopic biometric pattern. Material, Methods & Results: The right and left adrenal glands of 12 young (six females and six males), 12 adults (six females and six males), and 12 elderly (six females and six males), all mongrels, derived from routine and necropsied in the Animal Pathology Sector of the Veterinary Hospital of the Federal University of Espírito Santo (HOVET-UFES) were weighed and the length, width, and thickness were measured. For the microscopic measurement of the adrenal cortex, 10 randomly selected samples were submitted to routine histological processing and the microscope slides were observed under a light microscope at 5× and 10× objectives, photodocumented and measurements were obtained from a random portion of the adrenal cortex and the zones composing the cortex were measured in triplicate with the aid of the computerized image analysis software. The left adrenal showed a greater average length than the right adrenal in young, adult, and elderly dogs. The size of the glands in the young and adult dogs is not influenced by the sex of the animals, but in older dogs the females had a greater mean...

Juçara (Euterpe edulis Mart.) Supplementation Reduces Aberrant Crypt Foci and Increases SOD1 Expression in the Colorectal Mucosa of Carcinogenesis-Induced Rats.

Antioxidants present in food can act as a protective factor against the development of colorectal cancer (CRC) by reducing the development of aberrant crypt foci (ACF). This study aimed to analyze the effects of supplementation with juçara fruit pulp on the number of ACF and the SOD1 expression in an experimental model of CRC. Colorectal carcinogenesis was induced with 1,2-dimethylhydrazine (DMH) in 16 young female rats (Rattus norvegicus) given a diet supplemented with either juçara fruit pulp (DMH+/juçara+) or control (DMH+/juçara-). Five animals were used as a negative control (DMH-/juçara-). The (DMH+/juçara+) group received 14 days of supplementation (100 ml/animal/day) at 2-day intervals for 1 month. The number of ACF, area of positive staining for SOD1, and SOD1 expression score were evaluated. The (DMH+/juçara+) group presented a lower number of ACF, ACF > 3 crypts, and greater SOD1 expression in the colorectal mucosa. Based on the reduction in the number of lesions and possible positive impact on antioxidant enzymes, juçara fruit pulp appears to support the prevention of CRC, opening new possibilities for its use in dietary supplementation, as well as in the development of products and medications for the prevention and treatment of CRC.

PAI-1, CAIX, and VEGFA expressions as prognosis markers in oral squamous cell carcinoma.

BACKGROUND: In oral squamous cell carcinoma (OSCC), the HIF-1 complex promotes the expression of genes involved in specific mechanisms of cell survival under hypoxic conditions, such as plasminogen activator inhibitor-1 (PAI-1), carbonic anhydrase 9 (CAIX), and vascular endothelial growth factor A (VEGFA). The study aimed to investigate the presence and prognostic value of PAI-1, CAIX, and VEGFA in OSCC. MATERIALS AND METHODS: Immunohistochemistry was used to analyze the expressions of these proteins in 52 tumoral tissue samples of patients with OSCC, surgically treated and followed by a minimum of 24 months after surgery. The correlations between protein expressions and clinicopathological parameters and prognosis were analyzed. RESULTS: Positive PAI-1 membrane expression was significantly associated with local disease relapse (P = .027). Multivariate analysis revealed that the positive PAI-1 membrane expression is an independent marker for local disease relapse, with approximately 14-fold increased risk when compared to negative expression (OR = 14.49; CI = 1.40-150.01, P = .025). Strong PAI-1 cytoplasmic expression was significantly associated with the less differentiation grade (P = .027). Strong CAIX membrane expression was significantly associated with local disease-free survival (P = .038). Positive CAIX cytoplasmic expression was significantly associated with lymph node affected (P = .025) and with disease-specific survival (P = .022). Multivariate analysis revealed that the positive CAIX cytoplasmic expression is an independent risk factor for disease-related death, increasing their risk approximately 3-fold when compared to negative expression (HR = 2.84; CI = 1.02-7.87, P = .045). Positive VEGFA cytoplasmic expression was significantly associated with less differentiation grade (P = .035). CONCLUSION: Our results suggest a potential role for these expressions profiles as tumor prognostic markers in OSCC patients.

JMJD1A, H3K9me1, H3K9me2 and ADM expression as prognostic markers in oral and oropharyngeal squamous cell carcinoma.

AIMS: Jumonji Domain-Containing 1A (JMJD1A) protein promotes demethylation of histones, especially at lysin-9 of di-methylated histone H3 (H3K9me2) or mono-methylated (H3K9me1). Increased levels of H3 histone methylation at lysin-9 (H3K9) is related to tumor suppressor gene silencing. JMJD1A gene target Adrenomeduline (ADM) has shown to promote cell growth and tumorigenesis. JMJD1A and ADM expression, as well as H3K9 methylation level have been related with development risk and prognosis of several tumor types. METHODS AND RESULTS: We aimed to evaluate JMJD1A, ADM, H3K9me1 and H3K9me2expression in paraffin-embedded tissue microarrays from 84 oral and oropharyngeal squamous cell carcinoma samples through immunohistochemistry analysis. Our results showed that nuclear JMJD1A expression was related to lymph node metastasis risk. In addition, JMJD1A cytoplasmic expression was an independent risk marker for advanced tumor stages. H3K9me1 cytoplasmic expression was associated with reduced disease-specific death risk. Furthermore, high H3K9me2 nuclear expression was associated with worse specific-disease and disease-free survival. Finally, high ADM cytoplasmic expression was an independent marker of lymph node metastasis risk. CONCLUSION: JMJD1A, H3K9me1/2 and ADM expression may be predictor markers of progression and prognosis in oral and oropharynx cancer patients, as well as putative therapeutic targets.

Alterações clínicas e laboratoriais como indicadores para o tratamento anti-helmíntico em ovinos experimentalmente infectados com Haemonchus contortus / Clinical and laboratorial alterations as indicators for anti-helmintic treatment in sheep experimentally infected with Haemonchus contortus

A intensidade da anemia e hipoproteinemia são indicadores da gravidade da hemoncose. Este estudo objetivou verificar as alterações de peso, hematócrito e proteína plasmática, bem como a carga parasitária que indiquem a necessidade de intervenção anti-helmíntica em ovinos experimentalmente infectados com Haemonchus contortus. Foram utilizados 14 ovinos machos mestiços Santa Inês, 42,5 (±7,5) Kg, livres de nematoides, divididos em dois grupos contendo sete animais cada. Os animais do grupo infectado receberam 10.000 larvas de H. contortus cada. Todos os animais foram acompanhados aos 21, 28, 35, 42, 49, 56, 63 e 70 dias pós-infecção e avaliados segundo os parâmetros: OPG, Famacha©, peso, proteínas plasmáticas totais e hematócrito. No grupo infectado, o OPG variou e atingiu seu ápice aos 42 dias de acompanhamento, o hematócrito foi significativamente menor a partir da infecção experimental, o escore Famacha© variou de 1 a 2 e significativamente maior aos 21, 28, 35 e 42 dias, não houve alteração no nível de proteína plasmática nem no peso corpóreo em relação ao grupo controle. A infecção com até 10.000 larvas de H. contortus e OPG até 2500 não afeta significativamente o peso, hematócrito e proteína plasmática de ovinos mestiços Santa Inês, não havendo necessidade de intervenção anti-helmíntica.(AU)

The intensity of anemia and hypoproteinemia are indicators of the seriousness of haemonchosis. The objective of this study was to verify the changes in weight, hematocrit, and plasma protein, as well as the parasite load, indicating the need for anthelmintic intervention in sheep experimentally infected with Haemonchus contortus. Fourteen crossbred Santa Inês male sheep were used, weighing 42.5 (± 7.5) kg, nematodes-free. The animals were divided into two groups of seven animals each. The group of infected animals received 10,000 H. contortus larvae each. All animals were monitored after 21, 28, 35, 42, 49, 56, 63, and 70 days post-infection and the following parameters were assessed: EPG, Famacha©, weight, total plasma proteins, and hematocrit. In the infected group, the EPG varied and peaked on the 42nd follow-up day. The hematocrit became significantly lower after the experimental infection. Famacha score© ranged from 1 to 2 and it was significantly higher at 21, 28, 35 and 42 days. No change was observed in plasma protein level and body weight in the control group. Infection with up to 10,000 larvae of H. contortus. and EPG up to 2500 did not significantly affect weight, hematocrit, and plasma protein of crossbred Santa Inês sheep with no need for anthelmintic intervention.(AU)

ALTERAÇÕES CLÍNICAS E LABORATORIAIS COMO INDICADORES PARA O TRATAMENTO ANTI-HELMÍNTICO EM OVINOS EXPERIMENTALMENTE INFECTADOS COM Haemonchus contortus

Abstract The intensity of anemia and hypoproteinemia are indicators of the seriousness of haemonchosis. The objective of this study was to verify the changes in weight, hematocrit, and plasma protein, as well as the parasite load, indicating the need for anthelmintic intervention in sheep experimentally infected with Haemonchus contortus. Fourteen crossbred Santa Inês male sheep were used, weighing 42.5 (± 7.5) kg, nematodes-free. The animals were divided into two groups of seven animals each. The group of infected animals received 10,000 H. contortus larvae each. All animals were monitored after 21, 28, 35, 42, 49, 56, 63, and 70 days post-infection and the following parameters were assessed: EPG, Famacha©, weight, total plasma proteins, and hematocrit. In the infected group, the EPG varied and peaked on the 42nd follow-up day. The hematocrit became significantly lower after the experimental infection. Famacha score© ranged from 1 to 2 and it was significantly higher at 21, 28, 35 and 42 days. No change was observed in plasma protein level and body weight in the control group. Infection with up to 10,000 larvae of H. contortus. and EPG up to 2500 did not significantly affect weight, hematocrit, and plasma protein of crossbred Santa Inês sheep with no need for anthelmintic intervention.

Resumo A intensidade da anemia e hipoproteinemia são indicadores da gravidade da hemoncose. Este estudo objetivou verificar as alterações de peso, hematócrito e proteína plasmática, bem como a carga parasitária que indiquem a necessidade de intervenção anti-helmíntica em ovinos experimentalmente infectados com Haemonchus contortus. Foram utilizados 14 ovinos machos mestiços Santa Inês, 42,5 (±7,5) Kg, livres de nematoides, divididos em dois grupos contendo sete animais cada. Os animais do grupo infectado receberam 10.000 larvas de H. contortus cada. Todos os animais foram acompanhados aos 21, 28, 35, 42, 49, 56, 63 e 70 dias pós-infecção e avaliados segundo os parâmetros: OPG, Famacha©, peso, proteínas plasmáticas totais e hematócrito. No grupo infectado, o OPG variou e atingiu seu ápice aos 42 dias de acompanhamento, o hematócrito foi significativamente menor a partir da infecção experimental, o escore Famacha© variou de 1 a 2 e significativamente maior aos 21, 28, 35 e 42 dias, não houve alteração no nível de proteína plasmática nem no peso corpóreo em relação ao grupo controle. A infecção com até 10.000 larvas de H. contortus e OPG até 2500 não afeta significativamente o peso, hematócrito e proteína plasmática de ovinos mestiços Santa Inês, não havendo necessidade de intervenção anti-helmíntica.

Alterações clínicas e laboratoriais como indicadores para o tratamento anti-helmíntico em ovinos experimentalmente infectados com Haemonchus contortus / Clinical and laboratorial alterations as indicators for anti-helmintic treatment in sheep experimentally infected with Haemonchus contortus

A intensidade da anemia e hipoproteinemia são indicadores da gravidade da hemoncose. Este estudo objetivou verificar as alterações de peso, hematócrito e proteína plasmática, bem como a carga parasitária que indiquem a necessidade de intervenção anti-helmíntica em ovinos experimentalmente infectados com Haemonchus contortus. Foram utilizados 14 ovinos machos mestiços Santa Inês, 42,5 (±7,5) Kg, livres de nematoides, divididos em dois grupos contendo sete animais cada. Os animais do grupo infectado receberam 10.000 larvas de H. contortus cada. Todos os animais foram acompanhados aos 21, 28, 35, 42, 49, 56, 63 e 70 dias pós-infecção e avaliados segundo os parâmetros: OPG, Famacha©, peso, proteínas plasmáticas totais e hematócrito. No grupo infectado, o OPG variou e atingiu seu ápice aos 42 dias de acompanhamento, o hematócrito foi significativamente menor a partir da infecção experimental, o escore Famacha© variou de 1 a 2 e significativamente maior aos 21, 28, 35 e 42 dias, não houve alteração no nível de proteína plasmática nem no peso corpóreo em relação ao grupo controle. A infecção com até 10.000 larvas de H. contortus e OPG até 2500 não afeta significativamente o peso, hematócrito e proteína plasmática de ovinos mestiços Santa Inês, não havendo necessidade de intervenção anti-helmíntica.

The intensity of anemia and hypoproteinemia are indicators of the seriousness of haemonchosis. The objective of this study was to verify the changes in weight, hematocrit, and plasma protein, as well as the parasite load, indicating the need for anthelmintic intervention in sheep experimentally infected with Haemonchus contortus. Fourteen crossbred Santa Inês male sheep were used, weighing 42.5 (± 7.5) kg, nematodes-free. The animals were divided into two groups of seven animals each. The group of infected animals received 10,000 H. contortus larvae each. All animals were monitored after 21, 28, 35, 42, 49, 56, 63, and 70 days post-infection and the following parameters were assessed: EPG, Famacha©, weight, total plasma proteins, and hematocrit. In the infected group, the EPG varied and peaked on the 42nd follow-up day. The hematocrit became significantly lower after the experimental infection. Famacha score© ranged from 1 to 2 and it was significantly higher at 21, 28, 35 and 42 days. No change was observed in plasma protein level and body weight in the control group. Infection with up to 10,000 larvae of H. contortus. and EPG up to 2500 did not significantly affect weight, hematocrit, and plasma protein of crossbred Santa Inês sheep with no need for anthelmintic intervention.

Lavado vesical de bovinos com hematúria enzoótica: padronização de técnica de colheita, obtenção de amostras e avaliação citopatológica

ABSTRACT The bovine enzootic hematuria is a chronic disease that causes tumors in the bladder, and the cytophatologic test could assist in the early diagnosis. Aimed to standardize the technique of harvesting, obtaining samples and cytophatologic evaluation of bovine urinary bladder washing with bovine enzootic hematuria, 10 adult cows were divided into two groups: A (all the liquid infused in the bladder was recovered) and B (only the latter liquid was recovered). Liquids recovered were subjected to cytophatological evaluation. It was observed that the final volume of bladder liquid was higher in group A, however, in relation to the number of cells, no significant difference (p > 0.05) was observed between groups. The amount of inflammatory cells and epithelial cells obtained per sample revealed that in both groups all animals had more inflammatory cells than epithelial cells, however, there was no difference between the type of washing done. The epithelial cells were found in 60% of cases, the alterations observed were discretes and it was not possible to classify any sample as hyperplastic or neoplastic. Data from this study showed that the cytophatological examination of bovine urinary bladder washing may aid in the diagnosis of bovine enzootic hematuria and the two harvesting methods employed were adequate for obtaining viable samples. Cytopathological evaluation allowed the identification of non-neoplastic lesions predominantly inflammatory. It is believed that the use of molecular markers in cytological samples is important for early detection of pre-neoplastic or neoplastic lesions in these animals.

RESUMO A hematúria enzoótica bovina é uma doença crônica que causa neoplasias na bexiga, e o exame citopatológico poderia auxiliar no diagnóstico precoce. Objetivou-se padronizar a técnica de colheita, obtenção de amostras e avaliação citopatológica do lavado vesical de bovinos com hematúria enzoótica bovina. Foram utilizadas 10 vacas, adultas, distribuídas em dois grupos. No grupo A foi recuperado todo o líquido vesical infundido, no grupo B foi recuperado apenas o último lavado. Os lavados foram submetidos à avaliação citopatológica. Apesar do volume final de líquido vesical ter sido maior no grupo A, em relação ao número de células, não houve diferença significativa (p > 0,05) entre os grupos. A quantidade de células inflamatórias e células epiteliais obtidas por amostra revelou que nos dois grupos todos os animais apresentavam mais células inflamatórias do que epiteliais, entretanto, não houve diferença entre o tipo de colheita realizada. As células epiteliais foram encontradas em 60% dos casos e as alterações morfológicas observadas foram discretas, não sendo possível classificar nenhuma amostra como hiperplásica ou neoplásica. Os dados deste estudo permitiram concluir que o exame citopatológico do lavado vesical de bovinos pode auxiliar no diagnóstico da hematúria enzoótica bovina e que os dois métodos de colheita empregados mostraram-se adequados para obtenção de amostras viáveis. A avaliação citopatológica permitiu a identificação de lesões não neoplásicas predominantemente inflamatórias. Acredita-se que a utilização de técnicas moleculares com biomarcadores em amostras citológicas seria importante para detectar precocemente lesões pré-neoplásicas ou neoplásicas nesses animais.

Bovine urinary bladder washing with enzootic hematuria: standardization of harvesting technique, sample collection and cytopathological evaluation / Lavado vesical de bovinos com hematúria enzoótica: padronização de técnica de colheita, obtenção de amostras e avaliação citopatológica

The bovine enzootic hematuria is a chronic disease that causes tumors in the bladder, and the cytophatologic test could assist in the early diagnosis. Aimed to standardize the technique of harvesting, obtaining samples and cytophatologic evaluation of bovine urinary bladder washing with bovine enzootic hematuria, 10 adult cows were divided into two groups: A (all the liquid infused in the bladder was recovered) and B (only the latter liquid was recovered). Liquids recovered were subjected to cytophatological evaluation. It was observed that the final volume of bladder liquid was higher in group A, however, in relation to the number of cells, no significant difference (p  >  0.05) was observed between groups. The amount of inflammatory cells and epithelial cells obtained per sample revealed that in both groups all animals had more inflammatory cells than epithelial cells, however, there was no difference between the type of washing done. The epithelial cells were found in 60% of cases, the alterations observed were discretes and it was not possible to classify any sample as hyperplastic or neoplastic. Data from this study showed that the cytophatological examination of bovine  urinary bladder washing may aid in the diagnosis of bovine enzootic hematuria and the two harvesting methods employed were adequate for obtaining viable samples. Cytopathological evaluation allowed the identification of non-neoplastic lesions predominantly inflammatory. It is believed that the use of molecular markers in cytological samples is important for early detection of pre-neoplastic or neoplastic lesions in these animals.(AU)

A hematúria enzoótica bovina é uma doença crônica que causa neoplasias na bexiga, e o exame citopatológico poderia auxiliar no diagnóstico precoce. Objetivou-se padronizar a técnica de colheita, obtenção de amostras e avaliação citopatológica do lavado vesical de bovinos com hematúria enzoótica bovina. Foram utilizadas 10 vacas, adultas, distribuídas em dois grupos. No grupo A foi recuperado todo o líquido vesical infundido, no grupo B foi recuperado apenas o último lavado. Os lavados foram submetidos à avaliação citopatológica. Apesar do volume final de líquido vesical ter sido maior no grupo A, em relação ao número de células, não houve diferença significativa (p > 0,05) entre os grupos. A quantidade de células inflamatórias e células epiteliais obtidas por amostra revelou que nos dois grupos todos os animais apresentavam mais células inflamatórias do que epiteliais, entretanto, não houve diferença entre o tipo de colheita realizada. As células epiteliais foram encontradas em 60% dos casos e as alterações morfológicas observadas foram discretas, não sendo possível classificar nenhuma amostra como hiperplásica ou neoplásica. Os dados deste estudo permitiram concluir que o exame citopatológico do lavado vesical de bovinos pode auxiliar no diagnóstico da hematúria enzoótica bovina e que os dois métodos de colheita empregados mostraram-se adequados para obtenção de amostras viáveis. A avaliação citopatológica permitiu a identificação de lesões não neoplásicas predominantemente inflamatórias. Acredita-se que a utilização de técnicas moleculares com biomarcadores em amostras citológicas seria importante para detectar precocemente lesões pré-neoplásicas ou neoplásicas nesses animais.(AU)

Bovine urinary bladder washing with enzootic hematuria: standardization of harvesting technique, sample collection and cytopathological evaluation / Lavado vesical de bovinos com hematúria enzoótica: padronização de técnica de colheita, obtenção de amostras e avaliação citopatológica

The bovine enzootic hematuria is a chronic disease that causes tumors in the bladder, and the cytophatologic test could assist in the early diagnosis. Aimed to standardize the technique of harvesting, obtaining samples and cytophatologic evaluation of bovine urinary bladder washing with bovine enzootic hematuria, 10 adult cows were divided into two groups: A (all the liquid infused in the bladder was recovered) and B (only the latter liquid was recovered). Liquids recovered were subjected to cytophatological evaluation. It was observed that the final volume of bladder liquid was higher in group A, however, in relation to the number of cells, no significant difference (p  >  0.05) was observed between groups. The amount of inflammatory cells and epithelial cells obtained per sample revealed that in both groups all animals had more inflammatory cells than epithelial cells, however, there was no difference between the type of washing done. The epithelial cells were found in 60% of cases, the alterations observed were discretes and it was not possible to classify any sample as hyperplastic or neoplastic. Data from this study showed that the cytophatological examination of bovine  urinary bladder washing may aid in the diagnosis of bovine enzootic hematuria and the two harvesting methods employed were adequate for obtaining viable samples. Cytopathological evaluation allowed the identification of non-neoplastic lesions predominantly inflammatory. It is believed that the use of molecular markers in cytological samples is important for early detection of pre-neoplastic or neoplastic lesions in these animals.

A hematúria enzoótica bovina é uma doença crônica que causa neoplasias na bexiga, e o exame citopatológico poderia auxiliar no diagnóstico precoce. Objetivou-se padronizar a técnica de colheita, obtenção de amostras e avaliação citopatológica do lavado vesical de bovinos com hematúria enzoótica bovina. Foram utilizadas 10 vacas, adultas, distribuídas em dois grupos. No grupo A foi recuperado todo o líquido vesical infundido, no grupo B foi recuperado apenas o último lavado. Os lavados foram submetidos à avaliação citopatológica. Apesar do volume final de líquido vesical ter sido maior no grupo A, em relação ao número de células, não houve diferença significativa (p > 0,05) entre os grupos. A quantidade de células inflamatórias e células epiteliais obtidas por amostra revelou que nos dois grupos todos os animais apresentavam mais células inflamatórias do que epiteliais, entretanto, não houve diferença entre o tipo de colheita realizada. As células epiteliais foram encontradas em 60% dos casos e as alterações morfológicas observadas foram discretas, não sendo possível classificar nenhuma amostra como hiperplásica ou neoplásica. Os dados deste estudo permitiram concluir que o exame citopatológico do lavado vesical de bovinos pode auxiliar no diagnóstico da hematúria enzoótica bovina e que os dois métodos de colheita empregados mostraram-se adequados para obtenção de amostras viáveis. A avaliação citopatológica permitiu a identificação de lesões não neoplásicas predominantemente inflamatórias. Acredita-se que a utilização de técnicas moleculares com biomarcadores em amostras citológicas seria importante para detectar precocemente lesões pré-neoplásicas ou neoplásicas nesses animais.

Lavado vesical de bovinos com hematúria enzoótica: padronização de técnica de colheita, obtenção de amostras e avaliação citopatológica

ABSTRACT The bovine enzootic hematuria is a chronic disease that causes tumors in the bladder, and the cytophatologic test could assist in the early diagnosis. Aimed to standardize the technique of harvesting, obtaining samples and cytophatologic evaluation of bovine urinary bladder washing with bovine enzootic hematuria, 10 adult cows were divided into two groups: A (all the liquid infused in the bladder was recovered) and B (only the latter liquid was recovered). Liquids recovered were subjected to cytophatological evaluation. It was observed that the final volume of bladder liquid was higher in group A, however, in relation to the number of cells, no significant difference (p > 0.05) was observed between groups. The amount of inflammatory cells and epithelial cells obtained per sample revealed that in both groups all animals had more inflammatory cells than epithelial cells, however, there was no difference between the type of washing done. The epithelial cells were found in 60% of cases, the alterations observed were discretes and it was not possible to classify any sample as hyperplastic or neoplastic. Data from this study showed that the cytophatological examination of bovine urinary bladder washing may aid in the diagnosis of bovine enzootic hematuria and the two harvesting methods employed were adequate for obtaining viable samples. Cytopathological evaluation allowed the identification of non-neoplastic lesions predominantly inflammatory. It is believed that the use of molecular markers in cytological samples is important for early detection of pre-neoplastic or neoplastic lesions in these animals.

RESUMO A hematúria enzoótica bovina é uma doença crônica que causa neoplasias na bexiga, e o exame citopatológico poderia auxiliar no diagnóstico precoce. Objetivou-se padronizar a técnica de colheita, obtenção de amostras e avaliação citopatológica do lavado vesical de bovinos com hematúria enzoótica bovina. Foram utilizadas 10 vacas, adultas, distribuídas em dois grupos. No grupo A foi recuperado todo o líquido vesical infundido, no grupo B foi recuperado apenas o último lavado. Os lavados foram submetidos à avaliação citopatológica. Apesar do volume final de líquido vesical ter sido maior no grupo A, em relação ao número de células, não houve diferença significativa (p > 0,05) entre os grupos. A quantidade de células inflamatórias e células epiteliais obtidas por amostra revelou que nos dois grupos todos os animais apresentavam mais células inflamatórias do que epiteliais, entretanto, não houve diferença entre o tipo de colheita realizada. As células epiteliais foram encontradas em 60% dos casos e as alterações morfológicas observadas foram discretas, não sendo possível classificar nenhuma amostra como hiperplásica ou neoplásica. Os dados deste estudo permitiram concluir que o exame citopatológico do lavado vesical de bovinos pode auxiliar no diagnóstico da hematúria enzoótica bovina e que os dois métodos de colheita empregados mostraram-se adequados para obtenção de amostras viáveis. A avaliação citopatológica permitiu a identificação de lesões não neoplásicas predominantemente inflamatórias. Acredita-se que a utilização de técnicas moleculares com biomarcadores em amostras citológicas seria importante para detectar precocemente lesões pré-neoplásicas ou neoplásicas nesses animais.

Lavado vesical de bovinos com hematúria enzoótica: padronização de técnica de colheita, obtenção de amostras e avaliação citopatológica / Bovine urinary bladder washing with enzootic hematuria: standardization of harvesting technique, sample collection and cytopathological evaluation

A hematúria enzoótica bovina é uma doença crônica que causa neoplasias na bexiga, e o exame citopatológico poderia auxiliar no diagnóstico precoce. Objetivou-se padronizar a técnica de colheita, obtenção de amostras e avaliação citopatológica do lavado vesical de bovinos com hematúria enzoótica bovina. Foram utilizadas 10 vacas, adultas, distribuídas em dois grupos. No grupo A foi recuperado todo o líquido vesical infundido, no grupo B foi recuperado apenas o último lavado. Os lavados foram submetidos à avaliação citopatológica. Apesar do volume final de líquido vesical ter sido maior no grupo A, em relação ao número de células, não houve diferença significativa (p > 0,05) entre os grupos. A quantidade de células inflamatórias e células epiteliais obtidas por amostra revelou que nos dois grupos todos os animais apresentavam mais células inflamatórias do que epiteliais, entretanto, não houve diferença entre o tipo de colheita realizada. As células epiteliais foram encontradas em 60% dos casos e as alterações morfológicas observadas foram discretas, não sendo possível classificar nenhuma amostra como hiperplásica ou neoplásica. Os dados deste estudo permitiram concluir que o exame citopatológico do lavado vesical de bovinos pode auxiliar no diagnóstico da hematúria enzoótica bovina e que os dois métodos de colheita empregados mostraram-se adequados para obtenção de amostras viáveis. A avaliação citopatológica permitiu a identificação de lesões não neoplásicas predominantemente inflamatórias. Acredita-se que a utilização de técnicas moleculares com biomarcadores em amostras citológicas seria importante para detectar precocemente lesões pré-neoplásicas ou neoplásicas nesses animais.(AU)

The bovine enzootic hematuria is a chronic disease that causes tumors in the bladder, and the cytophatologic test could assist in the early diagnosis. Aimed to standardize the technique of harvesting, obtaining samples and cytophatologic evaluation of bovine urinary bladder washing with bovine enzootic hematuria, 10 adult cows were divided into two groups: A (all the liquid infused in the bladder was recovered) and B (only the latter liquid was recovered). Liquids recovered were subjected to cytophatological evaluation. It was observed that the final volume of bladder liquid was higher in group A, however, in relation to the number of cells, no significant difference (p > 0.05) was observed between groups. The amount of inflammatory cells and epithelial cells obtained per sample revealed that in both groups all animals had more inflammatory cells than epithelial cells, however, there was no difference between the type of washing done. The epithelial cells were found in 60% of cases, the alterations observed were discretes and it was not possible to classify any sample as hyperplastic or neoplastic. Data from this study showed that the cytophatological examination of bovine urinary bladder washing may aid in the diagnosis of bovine enzootic hematuria and the two harvesting methods employed were adequate for obtaining viable samples. Cytopathological evaluation allowed the identification of non-neoplastic lesions predominantly inflammatory. It is believed that the use of molecular markers in cytological samples is important for early detection of pre-neoplastic or neoplastic lesions in these animals.(AU)

HIF-1alpha expression profile in intratumoral and peritumoral inflammatory cells as a prognostic marker for squamous cell carcinoma of the oral cavity.

The HIF-1 transcriptional complex is responsible for controlling transcription of over 100 genes involved in cell hypoxia response. HIF-1alpha subunit is stabilized in hypoxia conditions, creating the HIF-1 nuclear transcription factor. In inflammatory cells, high HIF-1alpha expression induces lymphocytic immunosuppression, decreasing tumoral antigen recognition, which promotes tumor growth. The present work investigated the relationship between HIF-1alpha expression in lymphocytes populating the intratumoral and peritumoral region of 56 patients with oral cancer. Our data indicates a prognostic value for this expression. High HIF-1alpha expression in peritumoral inflammatory cells is significantly related to worse patient outcome, whereas high expression in the intratumoral lymphoid cells correlates with a better prognosis. A risk profile indicating the chance of disease relapse and death was designed based on HIF-1alpha expression in tumoral inflammatory cells, defining low, intermediate and high risks. This risk profile was able to determine that high HIF-1alpha expression in peritumoral cells correlates with worse prognosis, independently of intratumoral expression. Low HIF-1alpha in tumor margins and high expression in the tumor was considered a low risk profile, showing no cases of disease relapse and disease related death. Intermediate risk was associated with low expression in tumor and tumor margins. Our results suggest that HIF-1alpha expression in tumor and peritumoral inflammatory cells may play an important role as prognostic tumor marker.