Molecular docking, ADMET and molecular dynamics simulation revealed metralindole as a multitargeted inhibitor for division kinases / Ancoragem molecular, ADMET e simulação de dinâmica molecular revelaram o metralindol como inibidor multialvo para divisão quinases

Lung cancer is the most common type of cancer in the world, and alone, in 2020, almost 2.21 million new cases were diagnosed, with 1.80 million deaths, and are increasing daily. Non-small cell lung (NSCLC) is the primary type of lung cancer, predominantly forms around 80% of cases compared to small cell carcinoma, and about 75% of patients are already in an advanced state when diagnosed. Despite notable advances in early diagnosis and treatment, the five-year survival rate for NSCLC is not encouraging. Therefore, it is crucial to investigate the molecular causes of non-small cell lung cancer to create more efficient therapeutic approaches. Lung cancer showed a more significant and persistent binding affinity and energy landscape with the target CDK2 staurosporine and FGF receptor-1. In this study, we have picked two essential target proteins, human cyclin-dependent kinase-2 and Human Protein Kinase CK2 Holoenzyme and screened the entire prepared DrugBank prepared library of 1,55,888 compounds and identified 2-(2-methyl-5-nitroimidazole-1-yl) ethanol (Metralindole) as a major inhibitor. Metralindole has displayed high docking scores of -5.159 Kcal/mol and -5.99 Kcal/mol with good hydrogen bonding and other bonding topologies such as van der Waals force, and ADMET results shown excellent bioavailability, outstanding solubility, no side effects, and toxicity. The molecular dynamics simulation for 100ns in a water medium confirmed the compound's stability and interaction pattern with the lowest deviation and fluctuations. Our in-silico study suggests Metralindole, an experimental compound, can effectively cure lung cancer. Further, the experimental validation of the compound is a must before any prescription.

O câncer de pulmão é o tipo de câncer mais comum no mundo, e, apenas em 2020, foram diagnosticados quase 2,21 milhões de novos casos, com 1,8 milhão de óbitos, e isso vem aumentando diariamente. O câncer de pulmão de células não pequenas (CPCNP) é o tipo primário de câncer de pulmão, forma que predomina em cerca de 80% dos casos em comparação com o carcinoma de pequenas células, e cerca de 75% dos pacientes já estão em estado avançado quando diagnosticados. Apesar dos avanços notáveis no diagnóstico e tratamento precoces, a taxa de sobrevida em cinco anos para CPCNP não é animadora. Portanto, é crucial investigar as causas moleculares do CPCNP para criar abordagens terapêuticas mais eficientes. O câncer de pulmão mostrou uma afinidade de ligação e perfil energético mais significativos e persistentes com a estaurosporina CDK2 alvo e o receptor-1 de FGF. Neste estudo, escolhemos duas proteínas-alvo essenciais, a quinase dependente de ciclina humana-2 e a holoenzima CK2 da proteína quinase humana, examinamos toda a biblioteca preparada pelo DrugBank de 1,55,888 compostos e identificamos 2-(2-metil-5-nitroimidazol-1-il) etanol (metralindol) como inibidor principal. O metralindol apresentou altas pontuações de ancoragem de -5,159 Kcal/mol e -5,99 Kcal/mol, com boas ligações de hidrogênio e outras topologias de ligação, como a força de Van der Waals, e os resultados do ADMET mostraram excelente biodisponibilidade e solubilidade, sem efeitos colaterais e toxicidade. A simulação de dinâmica molecular para 100ns em meio aquoso confirmou a estabilidade e o padrão de interação do composto com os menores desvios e flutuações. Nosso estudo in silico sugere que o metralindol, um composto experimental, pode efetivamente curar o câncer de pulmão. Além disso, a validação experimental do composto é obrigatória antes de qualquer prescrição.

Primary pulmonary chondrosarcoma in a bitch

Background: Primary lung neoplasms are uncommon in veterinary medicine, and when they develop, they are more frequently observed to be of epithelial origin. Although chondrosarcomas are the second most diagnosed type of neoplasm in dogs at skeletal sites, their development in extraskeletal tissues, including the spleen, aorta, heart, tongue, peritoneum, and lungs, corresponds to approximately only 1% of cases. Therefore, the occurrence of primary pulmonary chondrosarcoma is considered very rare in domestic animals. Considering the rare occurrence and scarcity of data regarding its development, the present report describes the clinical and pathological aspects of a case of primary pulmonary chondrosarcoma in a bitch. Case: A 8-year-old mixed-breed bitch, weighing 14.2 kg, was examined at one private veterinary clinic with the principal complaint of prolonged respiratory distress and resistance to exercise. Clinical evaluation revealed tachypnea with expiratory dyspnea due to intense pleural effusion, tachycardia, and diffusely pale mucous membranes. Thoracocentesis was performed with drainage of 1000 mL of modified transudate, and fluid cytology, blood count, and chest radiography were performed. Cytological analysis of the thoracic fluid did not identify neoplastic cells, the blood count showed intense regenerative anemia, and the radiograph showed an extensive area of consolidation in the left caudal lung lobe, compatible with neoplasia. The results of the tests performed, in addition to the evolution of the clinical picture and the impossibility of performing the indicated surgical intervention, culminated in the patient's unfavorable prognosis, followed by euthanasia, necropsy, and histopathological evaluation of the collected material. Necropsy revealed a white neoplastic formation with reddish areas and firm consistency that diffused into the parenchyma of the left caudal lung lobe with invasion of the rib cage, fracture of the fifth and sixth left ribs, diaphragmatic metastasis, intense hydrothorax, and moderate hydroperitoneum. Microscopically, in the histological sections of the lung and diaphragm, poorly differentiated mesenchymal cells with moderate anisocytosis and anisokaryosis and interspersed with them, moderately differentiated chondrocytes surrounded by chondroid matrix, moderate anisocytosis and anisokaryosis, and a low mitotic index culminated in the diagnosis of primary pulmonary mesenchymal chondrosarcoma. Discussion: Malignant epithelial neoplasms were suspected; however, the histopathological features observed were compatible with primary pulmonary chondrosarcoma. The characteristics of the fluid collected through thoracentesis led to its classification as a modified transudate, which is often associated with neoplastic and hemorrhagic processes; both alterations were present in this case. The diagnosis of primary pulmonary chondrosarcoma was established based on the histopathological findings since the cell type and distribution observed in the present case were compatible with the typical pattern observed in extraskeletal chondrosarcomas. An unfavorable prognosis is common in cases of primary or metastatic lung neoplasm since, in most cases, the condition is identified in the advanced stages of the disease, making therapeutic management challenging. Given the increase in the number of dog deaths due to neoplasms in recent years, this case report may contribute to a better understanding of the biological behavior of pulmonary chondrosarcoma and assist in the choice of treatment to be adopted when required.

Glutamine ameliorates Bungarus multicinctus venom-induced lung and heart injury through HSP70: NF-κB p65 and P53/PUMA signaling pathways involved

Background: Bungarus multicinctus is one of the most dangerous venomous snakes prone to cardiopulmonary damage with extremely high mortality. In our previous work, we found that glutamine (Gln) and glutamine synthetase (GS) in pig serum were significantly reduced after Bungarus multicinctus bite. In the present study, to explore whether there is a link between the pathogenesis of cardiopulmonary injury and Gln metabolic changes induced by Bungarus multicinctus venom. We investigated the effect of Gln supplementation on the lung and heart function after snakebite. Methods: We supplemented different concentrations of Gln to mice that were envenomated by Bungarus multicinctus to observe the biological behavior, survival rate, hematological and pathological changes. Gln was supplemented immediately or one hour after the venom injection, and then changes in Gln metabolism were analyzed. Subsequently, to further explore the protective mechanism of glutamine on tissue damage, we measured the expression of heat-shock protein70 (HSP70), NF-κB P65, P53/PUMA by western blotting and real-time polymerase in the lung and heart. Results: Gln supplementation delayed the envenoming symptoms, reduced mortality, and alleviated the histopathological changes in the heart and lung of mice bitten by Bungarus multicinctus. Additionally, Gln increased the activity of glutamine synthetase (GS), glutamate dehydrogenase (GDH) and glutaminase (GLS) in serum. It also balanced the transporter SLC7A11 expression in heart and lung tissues. Bungarus multicinctus venom induced the NF-κB nuclear translocation in the lung, while the HO-1 expression was suppressed. At the same time, venom activated the P53/PUMA signaling pathway and the BAX expression in the heart. Gln treatment reversed the above phenomenon and increased HSP70 expression. Conclusion: Gln alleviated the glutamine metabolism disorder and cardiopulmonary damage caused by Bungarus multicinctus venom. It may protect lungs and heart against venom by promoting the expression of HSP70, inhibiting the activation of NF-κB and P53/PUMA, thereby delaying the process of snake venom and reducing mortality. The present results indicate that Gln could be a potential treatment for Bungarus multicinctus bite.

Acinar adenocarcinoma of the lung in a cat: case report / Adenocarcinoma acinar pulmonar primário em gato: relato de caso

Among the diseases that cause dyspnea in felines, primary pulmonary neoplasia is rare and tends to affect senile cats. This study reports the case of a seven teen year old FeLV infected cat who was diagnosed acinar adenocarcinoma of the lung and kidney metastasis. It presented prostration and anorexia and was hospitalized with dyspnea and pleural effusion. Chest radiography indicated increased radiopacity in the cranial portion of the right hemithorax, compatible with presence of intrathoracic mass and the cytologic analysis of pleural effusion suggested feline infectious peritonitis. The animal died two days after, and the definitive diagnosis was concluded after necropsy and histopathological examination. Pulmonary adenocarcinoma should be included with differential diagnosis of respiratory diseases in cats, especially the elderly.

Entre as doenças que causam dispneia em felinos, a neoplasia pulmonar primária é rara e costuma acometer gatos senis. Este trabalho tem por objetivo relatar o caso de um felino, de 17 anos, portador do vírus da leucemia felina, diagnosticado com adenocarcinoma acinar pulmonar primário com metástase no rim e na pleura parietal. O paciente apresentava histórico inespecífico, sendo as principais queixas prostração e anorexia, e foi internado apresentando dispneia e efusão pleural. A radiografia torácica indicou aumento da radiopacidade na porção cranial do hemitórax direito, compatível com presença de neoformação intratorácica. A citologia da efusão foi sugestiva de peritonite infecciosa felina. O animal veio a óbito dois dias após o atendimento inicial e o diagnóstico definitivo foi determinado por meio da necropsia e da avaliação histológica. O adenocarcinoma pulmonar deve ser incluído como diagnóstico diferencial de doenças respiratórias em gatos, principalmente idosos.

Circular RNA protein tyrosine kinase 2 aggravates pyroptosis and inflammation in septic lung tissue by promoting microRNA-766/eukaryotic initiation factor 5A axis-mediated ATP efflux

Purpose: Sepsis is characterized by an acute inflammatory response to infection, often with multiple organ failures, especially severe lung injury. This study was implemented to probe circular RNA (circRNA) protein tyrosine kinase 2 (circPTK2)-associated regulatory mechanisms in septic acute lung injury (ALI). Methods: A cecal ligation and puncture-based mouse model and an lipopolysaccharides (LPS)-based alveolar type II cell (RLE-6TN) model were generated to mimic sepsis. In the two models, inflammation- and pyroptosisrelated genes were measured. Results: The degree of lung injury in mice was analyzed by hematoxylin and eosin (H&E) staining and the apoptosis was by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling staining. In addition, pyroptosis and toxicity were detected in cells. Finally, the binding relationship between circPTK2, miR-766, and eukaryotic initiation factor 5A (eIF5A) was detected. Data indicated that circPTK2 and eIF5A were up-regulated and miR-766 was down-regulated in LPS-treated RLE-6TN cells and lung tissue of septic mice. Lung injury in septic mice was ameliorated after inhibition of circPTK2. Conclusion: It was confirmed in the cell model that knockdown of circPTK2 effectively ameliorated LPS-induced ATP efflux, pyroptosis, and inflammation. Mechanistically, circPTK2 mediated eIF5A expression by competitively adsorbing miR-766. Taken together, circPTK2/ miR-766/eIF5A axis ameliorates septic ALI, developing a novel therapeutic target for the disease.

Increased frequency of pneumonia in dogs with meningioma in ventral rhombencephalon / Aumento da frequência de pneumonia em cães com meningioma em rombencéfalo ventral

Intracranial tumors occurring in specific brain regions, such as the cerebellopontine angle, may be associated with cranial nerve dysfunction and dysphagia in humans and animals. Although dysphagia is a known risk factor for pneumonia, only postoperative pneumonia has been investigated in veterinary medicine. This study aimed to describe the clinical and pathological features of dogs with untreated intracranial meningiomas and concomitant pneumonia. Data from post-mortem examination registries from 2011 to 2021 were used (n=23). The frequency of pneumonia and other characteristics were compared between dogs with meningiomas in the ventral rhombencephalon region (VR group; n=13) and those with meningiomas in other intracranial sites (OIS group; n=10). The frequency of pneumonia was higher in the VR group than in the OIS group (n=5 vs. n=0; P=0.039). Plaque-like lesions were also more common in the VR group than in the OIS group (P=0.012). Dogs with concomitant pneumonia had cerebellopontine angle (n=3) and basilar meningiomas (n=2), mainly plaque-like lesions extending to or from other brain areas. In dogs with concomitant pneumonia, meningiomas had invasive (n=5) and compressive (n=3) growth behaviors and nerve roots involved in the swallowing process were frequently affected. Microscopically, these meningiomas were classified as atypical (n=4) and meningiomas (n=1). The reported clinical signs included anorexia (n=3), adipsia (n=1), and dysphagia (n=1). Our findings suggest untreated dogs with ventral rhombencephalon meningiomas may develop cranial nerve damage and aspiration pneumonia.

Tumores intracranianos que ocorrem em regiões específicas do cérebro, como o ângulo ponto-cerebelar, podem estar associados à disfunção de nervos cranianos e disfagia em humanos e animais. Embora a disfagia seja um conhecido fator de risco para pneumonia, apenas a pneumonia pós-operatória tem sido investigada na medicina veterinária. Este estudo teve como objetivo descrever as características clínicas e patológicas de cães com meningiomas intracranianos não tratados e pneumonia concomitante. Foram utilizados dados de registros de necropsias de 2011 a 2021. A frequência de pneumonia e outras características foram comparadas entre cães com meningiomas na região do rombencéfalo ventral (grupo VR; n=13) e aqueles com meningiomas em outros sítios intracranianos (grupo OIS; n=10). A frequência de pneumonia foi maior no grupo VR do que no grupo OIS (n=5 vs. n=0; P=0,039). Lesões tipo placa também foram mais comuns no grupo VR do que no grupo OIS (P=0,012). Cães com pneumonia concomitante apresentaram meningiomas no ângulo ponto-cerebelar (n=3) e região basilar (n=2), predominantemente lesões em forma de placa que se estendem de ou para outras áreas do cérebro. Em cães com pneumonia concomitante, os meningiomas apresentaram comportamentos de crescimento invasivo (n=5) e compressivo (n=3) e as raízes nervosas envolvidas no processo de deglutição foram frequentemente afetadas. Microscopicamente, esses meningiomas foram classificados como atípicos (n=4) e papilar (n=1). Os sinais clínicos relatados incluíram anorexia (n=3), adipsia (n=1) e disfagia (n=1). Nossos achados sugerem que cães com meningiomas no rombencéfalo ventral, não tratados, podem desenvolver lesão em nervos cranianos e pneumonia aspirativa.

Thoracoscopy pulmonary biopsy by two portals with a novel pre-tied loop ligature device in rabbits: an experimental study / Biópsia pulmonar por toracoscopia por dois portais com uma nova ligadura de alça pré-amarrada em coelhos: um estudo experimental

ABSTRACT: Thoracoscopy is replacing open lung biopsies because it is less invasive, usually the technique is done using three portals and intracorporeal suture technique. This study described the feasibility and efficacy of a novel pre tied loop ligature and to propose a thoracoscopic access strategy with two portals to perform lung biopsy in patients under 5 kg. Ten rabbits were positioned in dorsal recumbency. Total thoracoscopic lung biopsy was performed using a combined transdiaphragmatic approach and a right intercostal approach. A pre tied loop ligature was placed to perform a caudal lung lobe biopsy. Insufflation of the thoracic cavity was not performed. The total surgery time was 41.4 ± 14.5 min. The procedure was carried out free of complications that prevented slippage or tightening the knot or that made it come loose after the lung biopsy; there was no serious complication during the surgical procedure. The samples obtained averaged 1x0.64x0.45 cm (Length, Width, Depth) and were considered satisfactory according to the histopathologic evaluation. Thorax radiographs taken before and after the surgeries were compared and showed no pneumothorax or hemothorax. Necropsy confirmed no knot failure occurred at the biopsy site. The use of the novel pre tied loop ligature is a safe and effective technique, avoiding problems with the limited size of the thoracic cavity in small patients.

RESUMO: A toracoscopia está substituindo as biópsias pulmonares abertas por ser menos invasiva. Usualmente a técnica é feita utilizando três portais e ligaduras feitas intra corporalmente. O objetivo deste estudo foi descrever a viabilidade e eficácia de uma nova ligadura com alça pré-amarrada e propor uma estratégia de acesso toracoscópico com dois portais para realizar biópsia pulmonar em pacientes com menos de 5kg. Dez coelhos foram posicionados em decúbito dorsal. A biópsia pulmonar por toracoscopia total foi feita pela abordagem combinada transdiafragmática e abordagem intercostal direita. Uma nova ligadura pré-amarrada foi colocada para realizar uma biópsia do lobo pulmonar caudal. Não foi realizada insuflação da cavidade torácica. O tempo total de cirurgia foi de 41,4 ± 14,5 min. Os procedimentos foram realizados sem complicações que impedissem o escorregamento ou aperto do nó ou que o soltassem após a biópsia pulmonar; não houve nenhuma intercorrência grave durante o procedimento cirúrgico. As amostras coletadas tinham tamanho médio 1x0,64x0,45 cm e foram consideradas satisfatórias de acordo com a avaliação histopatológica. As radiografias de tórax feitas antes e depois das cirurgias foram comparadas e não mostraram pneumotórax ou hemotórax. A necropsia confirmou que não ocorreu falha do nó no local da biópsia. Conclui-se que uso da nova ligadura com alça pré-amarrada é uma técnica segura e eficaz, evitando problemas com o tamanho limitado da cavidade torácica em pacientes pequenos.

Effects of autolyzed yeast on leukocyte oxidative metabolism and pneumonia occurrence in feedlot steers / Efeito da levedura autolisada no metabolismo oxidativo leucocitário e na ocorrência de pneumonia em novilhos terminados em confinamento

Although yeast supplementation has shown an immunostimulant effect in feedlot cattle, the question remains whether it influences the occurrence of the bovine respiratory disease complex (BRD) in feedlot cattle. Therefore, our objective was to evaluate whether dietary supplementation with autolyzed yeast affects the efficiency of blood phagocytes, reducing inflammation and the occurrence of BRD in feedlot cattle. A randomized experimental trial was conducted with whole steers with half Angus blood for 106 days: control (n=10): diet without yeast; Y4 (n=12): yeast diet (4g per animal per day); and Y7 (n=7): yeast diet (7g per animal per day). On Days 0, 14, 42, 70, and 105, blood count, leukocyte oxidative metabolism, serum haptoglobin, frequency of nasal secretion and orbital temperature were evaluated. On slaughter day, a lung evaluation was performed. On Days 42 and 70 of feedlot finishing, the treated animals showed increased leukocyte oxidative metabolism (D42 P=0.04; D70 P=0.02) compared to the control group. These animals had a lower frequency of mucopurulent nasal secretion, lower orbital temperature and reduced pneumonia occurrence than the treated groups (P=0.05) with less lung lesion severity (P=0.01), allowing us to infer that the autolyzed yeast improves blood phagocytic function and decreases the inflammation and BRD of feedlot steers, especially at a dose of 4g.

Embora a suplementação com leveduras tenha demonstrado efeito imunoestimulante em bovinos, permanece-se a dúvida se ela influencia na ocorrência do complexo doença respiratória bovina (CDRB) em bovinos confinados. Portanto, objetivou-se avaliar se a suplementação dietética com levedura autolisada influencia a eficiência de fagócitos sanguíneos, reduzindo inflamações e a ocorrência de CDRB em bovinos confinados. Realizou-se um ensaio experimental randomizado com 36 novilhos inteiros 1/2 sangue Angus por 106 dias: controle (n=10): dieta sem levedura; Y4 (n=12): dieta com levedura (4g por animal por dia); e Y7 (n=7): dieta com levedura (7g por animal por dia). Nos dias 0, 14, 42, 70 e 105, hemograma, metabolismo oxidativo de leucócitos, haptoglobina sérica, frequência de secreção nasal e temperatura orbital foram avaliados. No dia do abate, foi realizada avaliação pulmonar. Nos dias 42, e 70 de terminação em confinamento, os animais tratados apresentaram aumento do metabolismo oxidativo dos leucócitos (D42 P=0,04; D70 P=0,02) em relação ao grupo controle. Esses animais apresentaram menor frequência de secreção nasal mucopurulenta, menor temperatura orbital, menor ocorrência de pneumonias (P=0,05) com menor gravidade das lesões pulmonares (P=0,01), permitindo inferir que a levedura autolisada na dieta melhorou a eficiência dos fagócitos sanguíneos, reduzindo inflamações e a ocorrência de CDRB, especialmente na dosagem de 4g.

Comparative evaluation of the efficacy of different marbofloxacin-based products in pigs at the nursery, growth and finish phases with clinical signs of respiratory disease / Avaliação comparativa da eficácia de diferentes produtos à base de marbofloxacina em suínos nas fases de creche, crescimento e terminação com sinais clínicos de doenças respiratórias

Marbofloxacin is one of the antibiotics of choice for the treatment of pigs with clinical manifestations of respiratory disease. Some commercial products on the market contain this active ingredient, and the objective of this study was to evaluate the efficacy of 3 available marbofloxacin-based products based on the SISAAB (single injection and short-term antibiotic) concept with regard to the performance and health parameters of pigs in the nursery, growth and finish phases with clinical manifestations of respiratory disease (RD). A total of 78 animals with clinical manifestations of RD from a total of 1726 pigs were used. The animals that had RD symptoms were weighed, identified and randomly subjected to 3 treatments: T1 - Forcyl® (n = 27); T2 - Marbox® (n = 25); T3 - Resolutor® (n = 26). All treatments corresponded to a single intramuscular dose of 8 mg of marbofloxacin per kg of live weight. After treatment, clinical signs of cough and depression, water consumption, rectal temperature, performance parameters, lung lesions and the pneumonia index of the animals were evaluated. The animals that received T1 and T3 maintained a mean rectal temperature of 39.38 °C 2 days after administration. In contrast, animals in the group that received T2 had significantly higher temperatures (p = 0.068) in this period (39.53 °C). There was a greater decrease in temperature (p = 0.042) in the 24 hours after drug administration in animals that received T1 (-0.573 °C). Regarding the other performance parameters, clinical signs of cough and depression, water consumption, lung injury and the pneumonia index, no differences were observed between the marbofloxacin-based products evaluated (p > 0.05). All marbofloxacin-based products were effective in reducing the clinical signs of RD and promoting animal performance and health. However, T1 was more effective in reducing the rectal temperature of the animals.

A marbofloxacina é um dos antibióticos selecionados para o tratamento de suínos com manifestações clínicas de doenças respiratórias. Existem alguns produtos comerciais no mercado com este princípio ativo, nesse sentido, objetivou-se com esse trabalho avaliar, com base no conceito SISAAB (antibioticoterapia de curta duração e dose única), a eficiência de três marbofloxacinas disponíveis sobre os parâmetros de desempenho e saúde de suínos em fase de creche, crescimento e terminação com quadros clínicos de doenças respiratórias (DR). Foram utilizados 78 animais com manifestações clínicas de DR de um total de 1726 porcos. Os animais que apresentaram sintomas de DR foram pesados, identificados e submetidos aleatoriamente a 3 tratamentos: T1 - Forcyl® (n = 27); T2 - Marbox® (n = 25); T3 - Resolutor® (n = 26). Todos os tratamentos corresponderam a uma dose única intramuscular de 8 mg de marbofloxacina por kg de peso vivo. Após o tratamento, foram avaliados os sinais clínicos de tosse e depressão, consumo de água, temperatura retal, parâmetros de desempenho, lesões pulmonares e o índice de pneumonia dos animais. Os animais que receberam T1 e T3 mantiveram temperatura retal média de 39,38°C, 2 dias após a administração. Em contrapartida, os animais do grupo que recebeu T2 apresentaram temperaturas significativamente mais elevadas (p < 0,10) neste período (39,53 °C). Em linha, foi possível observar a maior diminuição de temperatura (p = 0,042) nas 24 horas após a aplicação da medicação nos animais do grupo T1 (-0,573 °C). Em relação aos demais parâmetros de desempenho, sinais clínicos de tosse e estado de depressão, consumo de água, lesões pulmonares e índice de pneumonia, não foram observadas diferenças entre as marboloxacinas avaliadas (p>0,05). Todas as marbofloxacinas foram eficazes na redução dos sinais clínicos de doenças respiratórias, promovendo o desempenho e saúde dos animais, entretanto foi constatada a maior eficiência de T1 na redução da temperatura retal dos animais.

Thoracoscopy pulmonary biopsy by two portals with a novel pre-tied loop ligature device in rabbits: an experimental study / Biópsia pulmonar por toracoscopia por dois portais com uma nova ligadura de alça pré-amarrada em coelhos: um estudo experimental

Thoracoscopy is replacing open lung biopsies because it is less invasive, usually the technique is done using three portals and intracorporeal suture technique. This study described the feasibility and efficacy of a novel pre tied loop ligature and to propose a thoracoscopic access strategy with two portals to perform lung biopsy in patients under 5 kg. Ten rabbits were positioned in dorsal recumbency. Total thoracoscopic lung biopsy was performed using a combined transdiaphragmatic approach and a right intercostal approach. A pre tied loop ligature was placed to perform a caudal lung lobe biopsy. Insufflation of the thoracic cavity was not performed. The total surgery time was 41.4 ± 14.5 min. The procedure was carried out free of complications that prevented slippage or tightening the knot or that made it come loose after the lung biopsy; there was no serious complication during the surgical procedure. The samples obtained averaged 1x0.64x0.45 cm (Length, Width, Depth) and were considered satisfactory according to the histopathologic evaluation. Thorax radiographs taken before and after the surgeries were compared and showed no pneumothorax or hemothorax. Necropsy confirmed no knot failure occurred at the biopsy site. The use of the novel pre tied loop ligature is a safe and effective technique, avoiding problems with the limited size of the thoracic cavity in small patients.

A toracoscopia está substituindo as biópsias pulmonares abertas por ser menos invasiva. Usualmente a técnica é feita utilizando três portais e ligaduras feitas intra corporalmente. O objetivo deste estudo foi descrever a viabilidade e eficácia de uma nova ligadura com alça pré-amarrada e propor uma estratégia de acesso toracoscópico com dois portais para realizar biópsia pulmonar em pacientes com menos de 5kg. Dez coelhos foram posicionados em decúbito dorsal. A biópsia pulmonar por toracoscopia total foi feita pela abordagem combinada transdiafragmática e abordagem intercostal direita. Uma nova ligadura pré-amarrada foi colocada para realizar uma biópsia do lobo pulmonar caudal. Não foi realizada insuflação da cavidade torácica. O tempo total de cirurgia foi de 41,4 ± 14,5 min. Os procedimentos foram realizados sem complicações que impedissem o escorregamento ou aperto do nó ou que o soltassem após a biópsia pulmonar; não houve nenhuma intercorrência grave durante o procedimento cirúrgico. As amostras coletadas tinham tamanho médio 1x0,64x0,45 cm e foram consideradas satisfatórias de acordo com a avaliação histopatológica. As radiografias de tórax feitas antes e depois das cirurgias foram comparadas e não mostraram pneumotórax ou hemotórax. A necropsia confirmou que não ocorreu falha do nó no local da biópsia. Conclui-se que uso da nova ligadura com alça pré-amarrada é uma técnica segura e eficaz, evitando problemas com o tamanho limitado da cavidade torácica em pacientes pequenos.

Molecular identification of pathogenic Escherichia coli virulence genes and histopathological analysis in chickens condemned for airsacculitis / Identificação molecular de genes de virulência de Escherichia coli e análise histopatológica em frangos condenados por aerossaculite

ABSTRACT: The present study identified virulence genes and pathological changes caused by Escherichia coli in chicken carcasses condemned for airsacculitis and assessed if the histopathological examination and polymerase chain reaction (PCR) were effective for studies like this. Trachea, liver, and lung were collected from 30 chickens with suspected airsacculitis that has been condemned in the inspection line. The samples were analyzed by PCR to simultaneously identify two virulence genes (iss and tsh genes) and for histopathological testing. PCR efficiently genotypically characterize the E. coli isolates, where the virulence genes iss and tsh were found in three birds simultaneously. The histopathological examination detected a predominance of heterophils and mononuclear cells in the trachea (100%), lung (90%), and liver (13.3%). The liver was the organ where practically no alteration was diagnosed. The results of multiplex PCR for the tsh and iss virulence genes indicate the great potential of the approach in the characterization of E. coli isolates. Unspecific identification did not occur, thus making it necessary to use technologies for the identification and prevention of this agent in aviaries and poultry abattoirs.

RESUMO: O objetivo do presente estudo foi identificar genes de virulência e alterações patológicas provocadas por Escherichia coli em carcaças de frango condenadas por aerossaculite e se o exame histopatológico e uma Reação em Cadeia da Polimerase (PCR) são eficazes para os estudos dessa natureza. Para isso, foram coletados traqueia, fígado e pulmão de 30 frangos condenados na linha de inspeção, com suspeita de aerossaculite. As amostras foram submetidas a PCR para a identificação de dois genes de virulência de modo simultâneo (genes iss e tsh) e ao teste histopatológico. A PCR foi eficiente para caracterizar genotipicamente os isolados de E. coli, em que se constatou em três aves os genes de virulência iss e tsh simultaneamente. No exame histopatológico detectou-se a predominância de heterófilos e mononucleares na traqueia 100%, no pulmão 90% e no fígado 13,3%. O fígado foi o órgão onde praticamente não foi diagnosticada nenhuma alteração. Diante dos resultados obtidos, foi possível observar que a PCR multiplex para os genes de virulência tsh e iss apresenta um grande potencial na caracterização de isolados de E. coli, já que não gerou identificação inespecífica, com isso faz-se necessária a utilização de tecnologias para identificação e prevenção desse agente nos aviários e matadouros avícolas.

Real-time PCR quantification and histopathological findings of Mycoplasma hyopneumoniae infection in the lungs of pigs slaughtered in São Luís, Maranhão, Brazil / Quantificação por PCR em tempo real e achados histopatológicos da infecção por Mycoplasma hyopneumoniae em pulmão de suínos abatidos na cidade de São Luís, Maranhão, Brasil

ABSTRACT: Porcine enzootic pneumonia (PES), mainly caused by the bacteria Mycoplasma hyopneumoniae, is the main cause of respiratory problems in pigs. Infection by M. hyopneumoniae leads to production losses and the predisposition of affected animals to secondary infections, which may result in the condemnation of carcasses and organs due to lung lesions at the time of slaughter. The objective of the research was to evaluate the infection by M. hyopneumoniae in pigs submitted to slaughter in São Luís Island/MA, using molecular and histopathological diagnostic methods. One hundred fifty lung samples were collected from inspected (n=65) and non-inspected (n=85) slaughter pigs on São Luís Island, Maranhão, from July 2019 to August 2021. Of the 150 DNA samples collected, 121 showed an amplified product for Cyt B in the PCR assay. Thus, 121 samples were submitted to qPCR of M. hyopneumoniae, of which 44 (36.36%) showed positive results. The mean amount of bacterial load ranged from 1.20 × 101 to 7.20 × 104, with a mean of 1.73 × 104 copies. Of the reagent samples, 81.81% (36 samples) were obtained from non-inspected slaughter, while 18.18% (8 samples) were obtained from slaughterhouses. In the histopathological analysis, 44 positive qPCR samples were evaluated, of which 28 (63.63%) presented results compatible with the main inflammatory process associated with the presence of M. hyopneumoniae, that is, bronchial-associated lymphoid tissue hyperplasia (BALT). Three samples that showed the highest bacterial load (qPCR: 5.63 × 10³, 2.19 × 104 and 7.23 × 104) showed more evident lesions in this study. The microscopic findings associated with the quantifications indicated a relationship between the amount of bacterial load and the presence of microscopic lesions; higher bacterial load in lung tissue is associated with increased histopathologic staining for BALT hyperplasia. In conclusion, the results point to the circulation of the etiological agent in the sampled animals and the need for preventive measures on pig farms in Maranhão with the involvement of producers, sanitary defense and inspection agencies.

RESUMO: A pneumonia enzoótica suína (PES), causada principalmente pela bactéria Mycoplasma hyopneumoniae, é a principal causa de problemas respiratórios em suínos. A infecção por M. hyopneumoniae leva a perdas produtivas e a predisposição dos animais acometidos a infecções secundárias, o que pode resultar em condenação de carcaças e órgãos por lesões pulmonares no momento do abate. O objetivo da pesquisa foi avaliar a infecção por M. hyopneumoniae em suínos submetidos ao abate na Ilha de São Luís, por meio de métodos diagnósticos moleculares e histopatológicos. Para isso, foram coletadas 150 amostras de pulmão de suínos de abate inspecionado (n=65) e não inspecionado (n=85) na Ilha de São Luís/Maranhão, no período de julho de 2019 a agosto de 2021. Das 150 amostras de DNA coletadas, 121 apresentaram produto amplificado para Cyt B no ensaio de PCR. Assim, 121 amostras foram submetidas à qPCR de M. hyopneumoniae, das quais 44 (36,36%) apresentaram resultados positivos. A quantidade média de carga bacteriana variou de 1,20 × 101 a 7,20 × 104, com média de 1,73 × 104 cópias. Das amostras reagentes, 81,81% (36 amostras) foram obtidas de abate não inspecionado, enquanto 18,18% (8 amostras) foram obtidas em abatedouro. Na análise histopatológica, foram avaliadas 44 amostras positivas para qPCR, das quais 28 (63,63%) apresentaram resultados compatíveis com o principal processo inflamatório associado à presença de M. hyopneumoniae, ou seja, hiperplasia do tecido linfóide associado ao brônquio (BALT). Três amostras que apresentaram maior carga bacteriana (qPCR: 5,63 × 10³, 2,19 × 104 e 7,23 × 104) foram mais evidentes neste estudo. Os achados microscópicos associados às quantificações indicaram uma relação entre a quantidade de carga bacteriana e a presença de lesão microscópica; a maior carga bacteriana no tecido pulmonar está associada a maior alteração histopatológica para hiperplasia BALT. Em conclusão, os resultados obtidos sinalizam para a circulação do agente etiológico nos animais amostrados e a necessidade de medidas preventivas nas criações de suínos do estado do Maranhão com envolvimento dos produtores, órgãos de defesa sanitária e inspeção.

Probiotics supplementation reduces cigarette smoke-induced damage in the respiratory micro-architecture of mice / Suplementação com probióticos reduz os danos causados pela fumaça de cigarro na microarquitetura respiratória de camundongos

This study evaluated the protective efficacy of probiotics supplementation against cigarette smoke-induced lung emphysema, inflammation, and loss of cilia in mice. Probiotics are known to promote mucosal tolerance and mitigate respiratory injuries. Twenty-four adult mice were randomly divided into three groups: control (Ctr), cigarette smoke (CS), and cigarette smoke + probiotics (CS+P). Probiotics were given for 7 days before exposure to smoke in the CS+P group. Tissue samples of the trachea (goblet cell count and index, loss of cilia), lungs (airspace distention), and bronchoalveolar lavage fluid (BALF) were collected and processed. The results showed a significant increase in acidic and neutral goblet cells in the CS group compared to the Ctr and CS+P groups (P < 0.05). Overall, goblet cell number and index were lower in the CS+P group (41.71 ± 5.76, 0.67 ± 0.073) than CS group (56.28 ± 5.34, 1.31 ± 0.28). Inflammatory cells and loss of cilia significantly decreased in mice fed probiotics before exposure to cigarette smoke (P < 0.05). Lung emphysema was also significantly reduced in the CS+P group compared to the CS group (P < 0.05). In conclusion, dietary supplementation of probiotics reduced lung emphysema, inflammatory cells, goblet cell index, and loss of cilia under conditions of cigarette smoke exposure in mice.(AU)

O presente trabalho avaliou a eficácia da suplementação com probióticos contra a indução de enfisema, inflamação e perda de cílios por fumaça de cigarro em camundongos. Sabe-se que os probióticos promovem a tolerância da mucosa e mitigam as injúrias respiratórias. Vinte e quatro camundongos foram divididos, ao acaso, em três grupos: controle (CTR), fumaça de cigarro (CS) e fumaça de cigarro + probióticos (CS+P). Os probióticos foram fornecidos por sete dias antes da exposição à fumaça no grupo CS+P. Foram colhidas e processadas amostras de tecidos da traqueia (contagem de células caliciformes e index, perda de cílios), pulmões (distensão do espaço aéreo) e fluido de lavagem broncoalveolar. Os resultados obtidos revelaram um aumento significativo em células caliciformes acidificadas e neutralizadas no grupo CS quando comparado aos grupos Ctr e CS+P (P<0,05). Os números global e o índice de células caliciformes foram menores no grupo CS+P (41,71+ 5,76; 0,67+ 0,073) que no grupo CS (56,28+ 5,34 e 1,131+ 0,28). As células inflamatórias e a perda de cílios decresceram em camundongos alimentados com probióticos antes da exposição à fumaça de cigarro (P<0,05). O enfisema pulmonar também foi significativamente reduzido no grupo CS+P quando comparado ao grupo CS (P<0,05). A conclusão obtida foi que o fornecimento de dieta suplementada com probióticos reduziu o enfisema pulmonar, as células inflamatórias, o índice de células caliciformes e a perda de cílios nos camundongos expostos à fumaça de cigarro.(AU)

Threonine-to-lysine ratio in laying hens: physiological parameters and organ weight

The objective of this study was to evaluate the effect of threonine:digestible lysine ratio in the diet on the physiological variablesand weight of organs of light laying hens. Two hundred and ten 47 week-old Dekalb White laying henswere distributed in a completely randomized design, with five levels of threonine (0.507; 0.552; 0.597; 0.642 and 0.677%) and seven replicates of six birds each. The experimental period was 10 weeks, totaling 62 days and more eight days for the animals to adapt. The physiological parameters of cloacal temperature (CT), respiratory rate (RR) and average surface temperature (AST) were recorded weekly (7:00 am, 10:00 am, 1:00 pm, 4:00 pm, and 7:00 pm); after solid and water fasting,the birds were slaughtered to assess the absolute weight of the organs. The time of day influenced (p < 0.05) the physiological parameters RR and AST, and CT showed a significant effect (p < 0.05) of increasing levels of digestible threonine. The total weight of the pancreas, proventriculus and lung showed a significant effect (p < 0.05) of the increase in the levels of digestible threonine. The respiratory rate is affected by the levels of threoninein the diet. The 0.687% level promoted hypertrophy of the pancreas, proventriculus and lung, promoting more significant activity of these organs.(AU)

Caudal vena cava thrombosis in cows

Background: Caudal vena cava thrombosis (CVCT) is a serious disease that affects cattle. Due to being commonly a fatal pathology, it causes economic losses for producers and national livestock. Thus, the present study describes the epidemiological, clinical, ultrasonographic imaging and pathological findings in 4 cattle with CVCT attended at the Veterinary Hospital (HV) of the Institute of Veterinary Medicine (IMV) of the Federal University of Pará (UFPA). Cases: The animals were crossbreds of the Gir x Holstein and Jersey x Holstein breeds, aged between 4 and 8 years old, raised in semi-extensive and intensive systems. The main clinical signs were pale mucous membranes, reluctance to move, markedly positive venous pulse, engorged jugular with positive stasis test, and serous to mucopurulent nasal exudation. The auscultation of the lung fields revealed tachypnea, silent areas, wheezing, and pleural friction, in addition to coughing, expiratory dyspnea, mouth breathing, and expiratory grunts. One animal had severe hemoptysis. The ultrasound examination performed on a bovine revealed a circular and dilated caudal vena cava in cross-section. Laboratory tests in 3 cattle revealed anemia, leukocytosis with neutrophil left shift, and increased liver enzymes. At necropsy, all cattle had thrombi in the hepatic segment of the caudal vena cava. In the lung, multiple abscesses and areas of parenchymal consolidation, crateriform areas, as well as thrombi in the arteries were observed. Pleural effusion and ascites were seen in all cattle. Clotted blood was seen in the trachea, bronchi, and on rumen contents of an animal. Histopathological alterations seen in the liver were centrilobular hepatocytes with frequent intensely eosinophilic cytoplasm, and pyknotic, karyorrhexic, or absent nuclei and cell borders barely distinguishable. In the lung were nodular and random formations, with a thick wall of mature connective tissue and a central area full of cellular debris, necrotic cells, and intact and degenerated neutrophils (abscesses). Discussion: The set of diagnostic tools that include epidemiology, clinical signs and clinical examinations, ultrasound, necropsy, and histopathology were efficient in the diagnosis of CVCT. The possible causes that led the animals to develop CVCT were diffuse septic pododermatitis in the medial nail of the right pelvic limb associated with traumatic reticuloabomasitis and liver abscesses. In 1 cow, it was not possible to establish the probable cause of CVCT, but for the other cattle in the present study, the probable causes are in agreement with studies that have shown that this disease can occur as a sequel to several septic conditions such as jugular phlebitis, mastitis, hoof rot, enteritis, pneumonia, traumatic reticulopericarditis, acidosis and rumen laminitis, as well as omphalophlebitis in calves. The tachypnea, serous to purulent nasal exudation, pulmonary wheezing, pleural friction, coughing, and expiratory dyspnea, usually with open mouth breathing and expiratory grunts evidenced in the animals of this study, occurred as a result of embolic abscess pneumonia. The presence of multiple lung abscesses, areas of parenchymal consolidation, crateriform foci, and thrombi in the pulmonary arteries and chronic suppurative pneumonia, found at necropsy of the animals in the present study, are related to the development of a thrombus in the caudal vena cava that detaches and embolizes and lodges in the pulmonary arteries. The histopathological findings in 1 cow are compatible with lesions found at necropsy and draw attention to embolic pneumonia and liver lesions, which, are related to thrombi in pulmonary arteries and abscesses formed from CVCT, as well as venous stasis exerted in the return circulation.

Outbreak of leishmaniasis caused by Leishmania enriettii in guinea pigs (Cavia porcellus) / Surto de leishmaniose causada por Leishmania enriettii em cobaias (Cavia porcellus)

We describe an outbreak of leishmaniasis in seven guinea pigs (Cavia porcellus) in which nodular ulcerated skin lesions of varying sizes were observed in the nasal cavity, upper lip, pinnae, vulva, and periarticular region of the limbs. Cytologic exam of collected samples of the lesions in the auricle of one of the animals revealed macrophages containing parasitophorous vacuoles of approximately 4.0μm in diameter in their cytoplasm with morphology suggestive of Leishmania sp. Although skin lesions spontaneously regressed in two of the Guinea pigs, only one survived. All six animals that died were necropsied. Grossly, all animals showed bloody nodular cutaneous lesions with crusts. One of the guinea pigs had distended dark red and firm lungs. Histopathology of the skin lesions revealed histiocytic interstitial acanthotic dermatitis associated with a myriad of Leishmania organisms within macrophages cytoplasm. In the lung, the lesions were characteristic of broncho-interstitial pneumonia with focal infiltrates of neutrophils, epithelioid macrophages, and multinucleated giant cells containing 2µm basophilic amastigotes with morphology compatible with Leishmania spp. A focal granulomatous lesion ,associated with the causal agent in the lung is a novel description of leishmaniasis in guinea pigs caused by L. enriettii. The polymerase chain reaction (PCR) technique with mini-exon primer performed in samples of lesions from two affected guinea pigs was positive and equal to the reference strain, identifying Leishmania enriettii. The cytological, macroscopic, and histological lesions associated with the PCR technique allowed the diagnosis of leishmaniasis and the identification of the specie L. enriettii.

Descrevemos um surto de leishmaniose em sete cobaias (Cavia porcellus), com lesões cutâneas nodulares ulceradas de tamanhos variados observadas na cavidade nasal, lábio superior, pavilhões auriculares, vulva e região periarticular dos membros. No exame citológico foram encontrados macrófagos contendo vacúolos parasitóforos no citoplasma de aproximadamente 4.0μm em diâmetro com morfologia sugestiva de Leishmania sp. Apesar de regressão espontânea das lesões cutâneas terem ocorrido em duas das sete cobaias, apenas um sobreviveu. Seis dos sete animais afetados morreram e foram necropsiados. Macroscopicamente, todos os animais apresentaram lesões cutâneas nodulares, crostosas e sanguinolentas. Uma das cobaias tinha pulmões vermelho-escuros, distendidos e firmes. A histopatologia das lesões cutâneas revelou dermatite acantótica intersticial histiocítica associada a miríades de organismos de Leishmania no citoplasma de macrófagos. Nos pulmões as lesões eram características de pneumonia bronco-intersticial com infiltrado focal de neutrófilos, eosinófilos, macrófagos epitelioides e células gigantes multinucleadas contendo amastigotas basofílicos de 2µm com morfologia compatível com Leishmania spp. Lesões granulomatosas focais associadas ao agente no pulmão são um achado inédito na leishmaniose causada por L. enriettii em cobaias. A técnica de reação em cadeia da polimerase (PCR) com primer mini-exon realizada em amostras de lesões de duas cobaias afetadas foi positiva, identificando Leishmania enriettii. Os aspectos macroscópicos, citológicos, e histológicos associados à técnica da (PCR), permitiram o diagnóstico da leishmaniose e a identificação da espécie L. enriettii.

Effects of neutrophil extracellular traps during human respiratory syncytial virus infection in vitro

Abstract The human respiratory syncytial virus (hRSV) is the most common cause of severe lower respiratory tract diseases in young children worldwide, leading to a high number of hospitalizations and significant expenditures for health systems. Neutrophils are massively recruited to the lung tissue of patients with acute respiratory diseases. At the infection site, they release neutrophil extracellular traps (NETs) that can capture and/or inactivate different types of microorganisms, including viruses. Evidence has shown that the accumulation of NETs results in direct cytotoxic effects on endothelial and epithelial cells. Neutrophils stimulated by the hRSV-F protein generate NETs that are able to capture hRSV particles, thus reducing their transmission. However, the massive production of NETs obstructs the airways and increases disease severity. Therefore, further knowledge about the effects of NETs during hRSV infections is essential for the development of new specific and effective treatments. This study evaluated the effects of NETs on the previous or posterior contact with hRSV-infected Hep-2 cells. Hep-2 cells were infected with different hRSV multiplicity of infection (MOI 0.5 or 1.0), either before or after incubation with NETs (0.516 g/mL). Infected and untreated cells showed decreased cellular viability and intense staining with trypan blue, which was accompanied by the formation of many large syncytia. Previous contact between NETs and cells did not result in a protective effect. Cells in monolayers showed a reduced number and area of syncytia, but cell death was similar in infected and non-treated cells. The addition of NETs to infected tissues maintained a similar virus-induced cell death rate and an increased syncytial area, indicating cytotoxic and deleterious damages. Our results corroborate previously reported findings that NETs contribute to the immunopathology developed by patients infected with hRSV.

Resumo O vírus sincicial respiratório humano (hRSV) é a causa mais comum de doenças graves do trato respiratório inferior em crianças pequenas em todo o mundo, resultando em grande número de hospitalizações e gastos significativos para os sistemas de saúde. Neutrófilos são recrutados em massa para o tecido pulmonar de pacientes com doenças respiratórias agudas. No local da infecção, eles liberam armadilhas extracelulares de neutrófilos (NETs) que podem capturar e/ou inativar diferentes tipos de microrganismos, incluindo vírus. Evidências demonstraram que o acúmulo de NETs resulta em efeitos citotóxicos diretos nas células endoteliais e epiteliais. Os neutrófilos estimulados pela proteína F do vírus sincicial respiratório (hRSV-F) geram NETs que são capazes de capturar partículas virais, reduzindo assim sua transmissão. No entanto, a produção maciça de NETs obstrui as vias aéreas e aumenta a gravidade da doença. Assim, um maior conhecimento sobre os efeitos das NETs durante as infecções por hRSV é essencial para o desenvolvimento de novos tratamentos específicos e eficazes. Este estudo avaliou os efeitos das NETs no contato prévio ou posterior à infecção de células Hep-2 com hRSV. As células Hep-2 foram infectadas com diferentes quantidades de hRSV (multiplicidade de infecção ou MOI 0,5 ou 1,0), antes ou após a incubação com NETs (0,516 g/mL). Células infectadas e não tratadas mostraram redução da viabilidade celular e intensa coloração com azul de tripano, que foi acompanhada pela formação de sincícios numerosos e grandes. O contato prévio entre as NETs e as células não resultou em efeito protetor. As células em monocamadas mostraram um número e área de sincícios reduzidos, mas a morte celular foi semelhante àquela apresentada por células infectadas e não tratadas. A adição de NETs aos tecidos infectados manteve taxa de morte celular e formação de sincícios semelhantes àqueles induzidos pelo vírus em células não tratadas, indicando danos citotóxicos e deletérios. Nossos resultados corroboram achados relatados anteriormente de que as NETs contribuem para a imunopatologia desenvolvida por pacientes infectados com hRSV.

Carcinoma hepatocelular metastático em Amazona aestiva / Metastatic hepatocellular carcinoma in Amazona aestiva

Background: Hepatocellular carcinoma is a primary malignant tumor of the liver tissue and its occurrence in birds is considered rare. The tumor can occur as a single mass leading to hepatomegaly, or as multiple nodules in the liver. In animals of the genus Amazona, only 1 case of metastatic hepatocellular carcinoma has been reported in the United States, therefore, little is known about its epidemiology and clinicopathological aspects in these species. In this context, the aim of this work was to describe a case of metastatic hepatocellular carcinoma in an Amazona aestiva. Case: A blue-fronted amazon parrot (Amazona aestiva) was referred to necropsy after being found dead in its enclosure. On examination, it presented cachectic body score. Examination of the coelomic cavity, revealed a serous translucent fluid and adhesions between the liver and peritoneum.A red mass restricted to the right hepatic lobe and raised to the capsular surface, interspersed with whitish and dark red multifocal areas was observed. When cut, this mass was soft, protruding, multilobulated, whitish and with a friable reddish center. Additionally, on the dorsal surface of the left lung lobe, there was a rounded, well defined, whitish, and soft nodule. Microscopically, partial replacement of the hepatic parenchyma was observed by neoplastic proliferation of cuboidal epithelial cells, organized in mantle and supported by a scarce fibrovascular stroma. Cells have large, eosinophilic, well-delimited cytoplasm, with a central, oval nucleus, loose chromatin, and evident nucleolus. Moderate pleomorphism was characterized by anisocytosis, anisokaryosis, and aberrant nuclei. In the lung, a focally extensive mass with a pattern similar to that seen in the liver was observed. In the kidney, multifocal neoplastic emboli were noted. Liver immunohistochemistry was performed. Positive and negative controls were used to validate the reaction; however, there was no immunolabelling for the evaluated antibodies. Discussion: The histopathological characteristics observed in this study favored the diagnosis of hepatocellular carcinoma (HCC) with metastasis to kidney and lung. Primary liver tumors are rare in wild birds. In ducks, experimental studies have pointed aflatoxins and the duck hepatitis B virus as oncogenic agents, however, in birds of the genus Amazona, there are no studies that evaluate predisposing factors to the development of liver carcinoma. Macroscopically, hepatocellular carcinoma may present in massive, nodular or diffuse forms. In birds, the right lobe is the largest, which may suggest that this lobe is more prone to the development of HCC, as seen in the present case. The solid form, similar to that observed in this report, seems to be more commonly observed, as seen in the wild bird reports consulted. Metastases most often spread hematogenous, and in the present report there was metastasis to kidneys and lungs, which is a common feature for this neoplasm. In the present case, there was no labeling by any of the antibodies, perhaps because of their aggressiveness, associated with autolytic factors that prevent the labeling of antibodies, in addition to the specificity in the antibodyantigen relationship. This tumor must be differentiated from other liver tumors such as cholangiocarcinoma, and also the well-differentiated hepatocellular adenoma, in addition to non-neoplastic conditions. HCC should be considered as a differential diagnosis for Amazona aestiva found dead in the enclosure without previous clinical signs. This neoplasm is rare in Amazon parrots and reports should be encouraged in order to contribute to the understanding of the epidemiological and clinicopathological aspects of the tumor.

Plasma nitric oxide in dogs with pulmonary hypertension secondary or not to left-sided heart disease / Óxido nítrico plasmático em cães com hipertensão pulmonar secundária ou não à doença cardíaca esquerda

Nitric oxide (NO) is an important mediator responsible for vasodilation in pulmonary hypertension (PH) in humans. Based on human literature, it is suggested that in dogs there is also NO production decrease in lung tissue in the presence of PH with hypoxia. Therefore, the aim of this research was to determine the indirect plasmatic NO concentration in dogs with PH secondary or not to the left-side heart disease (LHD) and also with low, intermediate and high probability of PH to characterize the NO involvement on PH in dogs. Blood samples were collected from 35 dogs with probability of PH to NO measurement. NO concentration was estimated by the nitrite/nitrate concentration, and it was significantly different (p=0.002) in dogs with PH secondary to LHD (median=14 µM, range 11.19-16.59) and not secondary to LHD (median=25.88µM, range 15.08-36.71). However, this was not significant for the probability of low, intermediate, and high PH, although there was a tendency for NO concentration to be higher in dogs with high PH. The results of this study demonstrate that there is release of NO in dogs with PH, as well as that its dosage could differentiate dogs with PH secondary to LHD from dogs with non-secondary PH.

O óxido nítrico (ON) é um importante vasodilatador na hipertensão pulmonar (HP) em seres humanos. Baseado na literatura humana, sugere-se que em cães também ocorra a diminuição da produção de ON no tecido pulmonar na presença de HP frente à hipóxia. Dessa forma, o objetivo desta pesquisa foi determinar a concentração indireta do ON plasmático em cães com HP secundária ou não à doença do lado esquerdo do coração (LHD), bem como caracterizar os achados ecocardiográficos de diagnóstico e a probabilidade de HP (baixa, intermediária e alta). Para isso, foram coletadas amostras de sangue de 35 cães com probabilidade de HP para mensuração de ON. A concentração do ON foi estimada pela concentração de nitrito/nitrato, e esta foi significativamente diferente (P=0,002) nos cães com HP secundária à LHD (mediana=14µM, intervalo 11,19-16,59) e não secundária à LHD (mediana=25,88µM, intervalo 15,08-36,71), porém não significativa para a probabilidade de HP baixa, intermediária e alta, embora houvesse uma tendência a ser maior a concentração de ON nos cães com probabilidade alta. Os resultados deste estudo demonstram que há liberação do ON em cães com HP, bem como que sua dosagem conseguiu diferenciar cães com HP secundária à LHD de cães com HP não secundária.

Dirofilaria immitis parasite infection in cats: first case reported in the Legal Amazon region, Brazil: case report / Infecção por Dirofilaria immitis em gato: primeiro registro na região da Amazônia Legal, Brasil: relato de caso

A case of infection with Dirofilaria immitis in a cat is reported here with clinical signs of apathy, anorexia, dyspnea, polypnea, slight dehydration and pale mucus membranes. The radiographic examination showed cardiomegaly, lobar arteries dilation of the cranial lobes and tortuosity, enlargement of the caudal lobar arteries. In the right lobe of the lung parenchyma, interstitial pulmonary opacification tending to alveolar opacification was seen. The clinical signs, the movements, and the morphology of the microfilariae in the direct examination of fresh blood, peripheral blood smear and Knott's modified test supported the diagnosis. We alert to the need for clinicians to consider feline heartworm diseases as a differential diagnosis in endemic areas when cats show respiratory signs.

Descreve-se um caso de infecção por Dirofilaria immitis em gato com sinais clínicos de apatia, anorexia, dispneia, polipneia, leve desidratação e mucosas pálidas. O exame radiográfico demonstrou cardiomegalia, dilatação da artéria lobar cranial e tortuosidade, alargamento dos lobos caudais da artéria lobar. No lobo direito do parênquima pulmonar, opacificação pulmonar intersticial tendendo à opacificação foi observada. O diagnóstico foi baseado nos sinais clínicos, nos movimentos e na morfologia das microfilárias detectadas no exame de sangue a fresco, no esfregaço de sangue periférico e no teste de Knott modificado. Alertou-se para a necessidade de os clínicos considerarem a dirofilariose felina como diagnóstico diferencial em áreas endêmicas quando os gatos apresentam sinais respiratórios.