Presença da mutação do gene BRCA1 em cadelas com tumores mamários malignos / Presence of BRCA1 gene mutation in bitches with malignant mammary tumors

Mammary tumors (MTs) in bitches are similar to breast cancers in women. Thus, they can be used as a model for human breast cancer and findings can be extrapolated for use in human medicine. BRCA1 is a tumor suppressor gene. When the gene has a mutation, it cannot repair damaged DNA, which causes genetic instability and tumorigenesis. Therefore, we aimed to study the frequency of single nucleotide polymorphisms (SNPs) in the BRCA1 gene that are associated with distinct histological types of malignant MT in bitches. The study population consisted of 91 bitches, including a control group of 6 animals with healthy mammary glands and 85 animals with MTs. All animals underwent a presurgery evaluation consisting of a questionnaire administered to the person responsible for the animal, a physical examination, collection of peripheral blood for hematological and serum biochemistry evaluations, an electrocardiogram, and a preanesthesia evaluation. In addition, distant metastasis was studied via chest radiography and abdominal ultrasound. After evaluations were complete, the animals that could undergo surgery were administered general anesthesia and underwent a mastectomy or mammary gland sample collection. Histopathological examination and molecular analysis were performed to identify mutations in the BRCA1 gene. Histopathological examinations found 10 different types of malignant tumors in 36 sick animals. Tumor samples plus samples from the 6 control animals were subjected to DNA extraction, polymerase chain reaction (PCR) analysis, and genetic sequencing. The tumor with the highest incidence (33.33%) was a complex carcinoma, followed by carcinoma in mixed tumor (13.88), tubular carcinoma (13.88) and carcinosarcoma (13.88). Molecular analysis revealed 3 different SNP points in 5 samples (4006G>A, 3619A>G, and 3761C>T). The allelic variant 4006G>A (1/36) resulted in the alteration of the amino acid valine by isoleucine (V1336 I). The mutation 3619A>G (2/36) inserted the amino acid alanine instead of threonine (T1207 A). The mutation 3761C>T (2/36) led to the alteration of the amino acid serine by phenylalanine (S1254 F), a mutation for which there are no published reports. The histological types that showed BRCA1 mutations were complex carcinoma (1/5), carcinoma in mixed tumor (1/5), papillary carcinoma (1/5) and tubular carcinoma (2/5). Software analysis identified the new SNP (nucleotide 3761) in BRCA1 and 2 point mutations in nucleotides 4006 and 3619 and responsible for genetic instability. The development of breast cancer is caused by many endogenous and exogenous factors. The results of our study show that these factors have a greater presence in female, mixed breed, uncastrated, and older dogs, confirming the data in the veterinary literature. In the present study, we found different histological types of malignant breast tumors with mutations in the BRCA1 gene, as other authors have reported. However, we also found the mutation 3761C>T, which, to the best of our knowledge, has not been reported in the literature. This shows the need for studies in veterinary medicine that assess mutations in the BRCA1 gene and the most common histological types. In conclusion, SNPs in the BRCA1 gene cause genetic instability, resulting in additional mutations that lead to the development of breast tumors. They are point mutations that affect transcription, resulting in truncated proteins. These proteins may have a loss of function, leading to carcinogenesis.(AU)

Polymerase Chain Reaction and blood culture for diagnosis of canine sepsis / Reação em Cadeia da Polimerase e hemocultura para diagnóstico de sepse canina

ABSTRACT: Sepsis is characterized by the presence of organ dysfunction secondary to the dysregulated systemic inflammatory response associated with an infection, and has high mortality rates. Traditional diagnostic techniques based on non-microbiological isolation are time-consuming and may delay treatment. Thus, this study aimed to compare bacterial and fungal broad-range polymerase chain reaction (PCR) and blood culture for diagnosis of sepsis in dogs. Blood samples from 88 dogs with suspected sepsis were analyzed by blood culture, and PCR to detect bacterial and fungal DNA. On blood culture, 20 (22.7%) samples tested positive for bacterial isolates; however, none tested positive for fungi. Through PCR analysis, bacterial DNA was detected in 46 (52.3%) animals, whereas fungal DNA was present in one (1.1%) sample. Our results showed that PCR-based testing has important diagnostic value for canine blood infections because it has a shorter turnaround time and higher sensitivity than traditional blood culture.

RESUMO: Sepse se caracteriza pela presença de disfunção orgânica secundária à resposta inflamatória sistêmica desregulada, associada a uma infecção com elevadas taxas de mortalidade. As técnicas tradicionais baseadas no isolamento microbiológico são demoradas e podem atrasar o tratamento. O objetivo deste estudo foi comparar a Reação em Cadeia da Polimerase (PCR) bacteriana e fúngica e hemocultura em cães com sepse. Foram analisadas 88 amostras de sangue de cães com suspeita de sepse por meio de hemocultura e PCR para detectar DNA bacteriano e fúngico. Nas culturas sanguíneas, 20 (22,7%) amostras foram positivas para isolados bacterianos. No entanto, nenhuma amostra foi positiva para fungos. Através da análise por PCR, o DNA bacteriano foi detectado em 46 animais (52,3%), enquanto que o DNA fúngico estava presente em uma amostra (1,1%). Neste caso, a PCR apresenta importante valor diagnóstico em cães com infeções sanguíneas devido a sua rapidez e maior sensibilidade do que a isolamento por hemocultura.

Síntese e caracterização de nanopartículas de ouro conjugadas com curcumina e seus efeitos na osteoartrite experimental induzida / Synthesis and characterization of gold nanoparticles combined with curcumin and its effects on experimentally induced osteoarthritis

RESUMO: A Osteoartrite (OA) é uma denominação clínica para uma combinação de condições patológicas que envolvem a degeneração progressiva da cartilagem articular e remodelação de osso subcondral. A curcumina, um potente agente anti-inflamatório, têm sido extensivamente estudada, no entanto não oferece boa biodisponibilidade sistêmica. Nanopartículas de ouro (AuNPs) apresentam aplicações potenciais na administração de substâncias terapêuticas aumentando a eficiência do transporte de fármacos. O objetivo deste estudo foi realizar a síntese e caracterização de um sistema conjugando as AuNPs à curcumina e avaliar seu potencial terapêutico em um modelo experimental de OA em camundongos por desestabilização do menisco medial (DMM). As AuNPs foram conjugadas com curcumina e os sistemas foram caracterizados por espectroscopia no UV-VIS, espalhamento de luz dinâmico (DLS) e determinação do potencial zeta. Formou-se 4 grupos de oito animais cada, denominados A, B, C, D que receberam injeção intra-articular de AuNPs, curcumina, AuNP-curcumina e solução fisiológica, respectivamente. Após 7 semanas, a cartilagem da articulação-femoro- tibio-patelar (AFTP) foi avaliada em uma variação de escore de 0 a 24. A conjugação de AuNP-curcumina mostrou boa estabilidade e aplicação terapêutica, mas não diferiu significativamente (P>0,05) dos grupos A e B, no entanto, mostrou menor valor de escore e significância (P<0,001) em relação ao grupo controle. Os resultados deste trabalho mostram a importância do desenvolvimento de novos nanofármacos. Neste caso a conjugação de AuNPs com a curcumina permitiu a obtenção de um nanofármaco com sugestivo potencial para aplicação no tratamento da OA.

ABSTRACT: Osteoarthritis (OA) is the clinical term for a combination of pathological conditions that involve the progressive degeneration of articular cartilage and subchondral bone remodelling. Curcumin, a potent anti-inflammatory agent, has been extensively studied; however, it does not provide good systemic bioavailability. Gold nanoparticles (AuNPs) have potential applications in the administration of therapeutic substances in order to increase the transport efficiency of drugs. The objectives of this study were to explore the synthesis and characterization of a system combining AuNPs with curcumin and evaluate its therapeutic potential in an experimental model of OA in mice by the destabilization of the medial meniscus (DMM). The AuNPs were conjugated with curcumin and the systems were characterized by UV-VIS spectroscopy, dynamic light scattering (DLS), and zeta potential. Four groups of eight animals each were formed and labelled A, B, C, and D, which received intra-articular injections of AuNPs, curcumin, AuNP-curcumin, and physiologic solution, respectively. After seven weeks, the cartilage of the stifle joint (SJ) was rated on a scale ranging from 0 to 24. Combination of AuNP-curcumin demonstrated good stability and therapeutic applications, but it did not differ significantly (P>0.05) from groups A and B. However, the control group had a significantly lower score (P<0.001). Results of this study demonstrated the importance of developing new nanodrugs. In this case, the combination of AuNPs and curcumin yielded the nanodrug effects suggestive of a potential for application in the treatment of OA.

Ultrasound characterization of the coelomic cavity organs of the red-footed tortoise ( Chelonoidis carbonaria ) / Caracterização ultrassonográfica dos órgãos da cavidade celomática do jabutipiranga (Chelonoidis carbonaria)

ABSTRACT: Herein it was describe sonographic morphology and sintopy of the bowels of the coelomic cavity in the red-footed tortoise. Coelomic cavity of 19 males and 19 females were scanned through cervical and prefemoral access with a multifrequency sector transducer. Morphology, syntopy and echogenicity of the heart, thyroid, liver, gallbladder, reproductive organs, stomach, small intestine, large intestine, urinary bladder and kidneys were evaluated. The heart showed two atria and one ventricle with a thick, trabecular wall. The thyroid was oval and hyperecoic, visualized in the cardiac portion of the ultrasound. The liver, gallbladder and digestive system were similar to those seen in mammals and turtles. However, the tortoise liver was relatively more hyperechoic than mammals. The kidneys appeared as triangular structures, which were hypoechoic, homogeneous and vascularized; the bladder was observed mostly as being elongated with anechoic content, and its wall appeared as a thin hyperechoic line when free fluid was present. The testes were observed to be elongated, homogeneous and more hyperechoic than kidneys. The ovarian follicles were seen as hyperechoic, echogenic balls of variable size and quantity, the oviduct as a sigmoid tubular structure and the eggs as thin hyperechoic lines with posterior acoustic shadowing. In some animals, there were variable amounts of fluid around the heart and in the coelomic cavity.

RESUMO: Neste trabalho é descrita a morfologia ecográfica e sintopia das vísceras da cavidade celomática do jabutipiranga. Foram examinados 19 machos e 19 fêmeas por meio de acesso cervical e pré-femoral com um transdutor setorial multifrequencial. Foram avaliadas morfologia, sintopia e ecogenicidade do coração, tiróide, fígado, vesícula biliar, órgãos reprodutivos, estômago, intestino delgado, intestino grosso, bexiga urinária e rins. O coração mostrou dois átrios e um ventrículo com uma parede trabecular e espessa. Na base cardíaca pode-se visibilizar a tireóide oval e ecogênica. O fígado, vesícula biliar e sistema digestivo foram semelhantes ao observado em mamíferos e testudinos. No entanto, o fígado dos quelônios mostrou-se mais hiperecóico em relação ao dos mamíferos. Os rins foram vistos como estruturas triangulares, hipoecóicas, homogêneas e vascularizadas; a bexiga foi observada na maior parte como alongada com conteúdo anecóico, e sua parede foi vista como uma fina camada hiperecóica. Os testículos foram observados como estruturas alongadas, homogêneas e hiperecóicas em relação aos rins. Os folículos ovarianos foram vistos como esferas ecogênicas a hiperecogênicas de tamanho e quantidade variável, o oviduto como uma estrutura tubular sigmóide e os ovos como finas linhas hiperecogênicas com sombra acústica posterior. Em alguns animais, havia quantidades variáveis de líquido em volta do coração e na cavidade celomática.

Pythium insidiosum colitis in a dog: treatment and clinical outcome / Colite por Pythium insidiosum em um cão: tratamento e evolução clínica

ABSTRACT: The aim of this report is to describe the clinical, pathological and imaging findings and treatment of colitis caused by Pythium insidiosum in a canine presenting haematochezia and progressive weight loss. Through imaging, a thickening of the transverse and descending colon was observed. Histopathological analysis of the large intestine fragment revealed the presence of hyphae, confirmed by immunohistochemistry and PCR as P. insidiosum. Antifungal treatment with itraconazole implemented after partial surgical resection, resulted in control of the disease.

RESUMO: O objetivo deste relato é descrever os achados clínico-patológicos, de imagem e o tratamento de colite por Pythium insidiosum em canino apresentando hematoquezia e emagrecimento progressivo. Nos exames de imagem, foram observados espessamento do cólon transverso e descendente. Análise histopatológica de fragmento do intestino grosso revelou a presença de hifas, confirmado pela imuno-histoquímica e PCR como P. insidiosum. Terapia antifúngica com itraconazol foi instituída pós-ressecção cirúrgica parcial, obtendo-se controle da doença.

Evaluation of intraluminal pressure in cystorrhaphies with and without intestinal serosal patch supplementation from canine cadavers

PURPOSE:To evaluate the maximal intraluminal pressure (MIP) supported by canine cadaveric urinary bladders that underwent cystotomy followed by cystorraphy, with and without serosal patching-supplementation.METHODS:Two groups (n=8 each) were formed, and in one (conventional) the cystotomy was closed with cushing pattern. In the other group (serosal), the same procedure was performed, and a piece of jejunum was used for the construction of the serosal patching over the cystorraphy. MIP was measured by means of an invasive blood pressure transducer with closed stopcock attached to a multiparameter monitor. At the end of each measurement, the bladder body circumference was assessed.RESULTS:Mean±SD MIP sustained for the conventional and serosal groups were 28.88±5.08 and 65.38±10.99 mmHg, respectively (p<0.0001). Bladder circumference did not change significantly between groups (p=0.35) and did not correlate with MIP assessed in conventional (p=0.27; r=0.4379) and serosal groups (p=0.37; r=-0.3637).CONCLUSION:Serosal patch-supplemented cystorraphies were able to sustain intraluminal pressures 55.8% higher, than nonsupplemented cystorraphies in specimens from canine cadavers.

Experimental osteoarthritis in rabbits: lesion progression / Osteoartrite experimental em coelhos: progressão lesional

The aim of this study was to evaluate the progression of lesions in different stages of osteoarthritis (OA) experimental by radiography (RX), computed tomography (CT), macroscopic and histopathology, linking these different diagnostic methods, helped to provide information that helps the best time for the therapeutic approach. Four experimental periods were delineated at 3, 6, 9 and 12 weeks after induction of OA, known as PI, PII, PIII and PIV, respectively, each with six animals. We evaluated the five compartments of the femorotibial joint: medial femoral condyle (MFC), lateral femoral condyle (LFC), medial tibial plateau (MTP), lateral tibial plateau (LTP) and femoral trochlea (FT). Therefore we established an index by compartment (IC) and by adding such an index was estimated joint femorotibial (IFT). It was observed that the CFM was the compartment with the highest IC also differed significantly (p<0.05) from other compartments. Compartments showed no significant difference (p>0.05) between the PI and PII, however contrary fact occurred between the PII and PIII (p<0.05), PIII and PIV (p<0.01) and between PI and PIV (p<0.001). Similarly the IFT, showed a significant difference in the animals of PIV compared to PI (p<0.001), PII (p<0.001) and PIII (p<0.01), and there was no statistical difference (p> 0.05) between the PI and PII. In the variation of the average interval between periods, there was a higher value between the PIII PIV and for the other intervals of time periods (PI, PII, and PIII-PII). However, these intervals showed no statistically significant difference (p>0.05). Through the RX, CT, macroscopic and histopathological findings, we found similar patterns among individuals within the same period demonstrating a gradual progression of the disease. These results show that between 3 and 6 weeks progression of the lesion is slower and probably also can be reversed in comparison to other ranges where proved further progression between 9 and 12 weeks after induction of trauma OA. These results may provide a better therapeutic approach aimed at reversing the lesions in early stages of OA. We conclude that the interconnection of the four diagnostic methods individually classified into scores, which were unified in both indices in the evaluation by the femorotibial joint compartment and may represent a diagnostic condition closer to the true condition of the injury and its progression.

O objetivo deste estudo foi avaliar a progressão das lesões em diferentes períodos da osteoartrite (OA) experimental em coelhos, através da radiografia (RX), tomografia computadorizada (TC), macroscopia e histopatologia, interligar estes diferentes métodos diagnósticos, bem como trazer informações que ajudem na decisão do melhor momento para a abordagem terapêutica. Foram delineados quatro períodos experimentais às 3, 6, 9 e 12 semanas após a indução da OA, denominados como PI, PII, PIII e PIV, respectivamente, cada qual com seis animais. Foram avaliados os cinco compartimentos da articulação femorotibial: côndilo femoral medial (CFM), côndilo femoral lateral (CFL), platô tibial medial (PTM), platô tibial lateral (PTL) e tróclea femoral (TF). Por conseguinte, estabeleceu-se um índice por compartimento (IC) e através da soma destes foi obtido um índice da articulação femorotibial (IFT). Observou-se que, o CFM foi o compartimento com maior valor de IC, além disso, diferiu significativamente (p<0,05) dos demais compartimentos. Os compartimentos não apresentaram diferença significativa (p>0,05) entre o PI e o PII, no entanto fato contrário ocorreu entre o PII e o PIII (p<0,05), PIII e PIV (p<0,01) e entre o PI e PIV (p<0,001). Similarmente o IFT, apresentou diferença significativa nos animais do PIV em relação ao PI (p<0,001), ao PII (p<0,001) e ao PIII (p<0,01), e não houve diferença estatística (p>0,05) entre o PI e PII. Na variação média dos intervalos entre os períodos, observou-se um maior valor entre o PIII e o PIV em relação aos outros intervalos de períodos (PI-PII e PII-PIII). No entanto, estes intervalos não apresentaram diferença estatisticamente significativa (p>0.05). Através do RX, TC, macroscopia e histopatológico, verificamos padrões similares entre os indivíduos dentro do mesmo período demonstrando assim uma gradual progressão da doença. Tais resultados evidenciam que entre as 3 e 6 semanas a progressão da lesão é mais lenta e muito provavelmente ainda pode ser reversível em comparação aos outros intervalos onde se comprovou uma maior progressão entre as 9 e 12 semanas após a indução traumática da OA. Estes resultados podem propiciar uma melhor abordagem terapêutica objetivando a reversão das lesões em fases iniciais da OA. Concluímos que, a interligação dos quatro métodos diagnósticos, individualmente classificados em escores e que foram unificados em índices tanto na avaliação por compartimento quanto pela articulação femorotibial pode representar uma condição diagnóstica mais próxima à verdadeira condição da lesão quanto de sua progressão.