Isoliquiritigenin regulates the circ_0002860/miR-431-5p/RAB9A axis to function as a tumor inhibitor in melanoma

Background: Isoliquiritigenin (ISL) presents antitumor effects against melanoma cells. It is known that various circular RNAs (circRNAs) are involved in the development of melanoma. Therefore, the present study aims to investigate the molecular mechanisms of ISL and circ_0002860. Methods: Circ_0002860, microRNA-431-5p (miR-431-5p) and member RAS oncogene family (RAB9A) were detected through reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay. Cell viability was examined via cell counting kit-8 assay. The proliferation ability was assessed using colony formation assay. Cell apoptosis and cell cycle were determined by flow cytometry. Transwell assay was used for detection of migration and invasion. Western blot was conducted for protein analysis. Target binding was confirmed via dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. In vivo research was performed through xenograft tumor assay. Results: Circ_0002860 was downregulated by ISL in melanoma cells. ISL-induced inhibitory effects on cell proliferation, cell cycle progression, migration and invasion were alleviated by circ_0002860 overexpression. MiR-431-5p was a target of circ_0002860. Circ_0002860 eliminated the ISL-induced tumor inhibition via sponging miR-431-5p in melanoma cells. Circ_0002860 elevated the RAB9A level by targeting miR-431-5p. The function of ISL was related to miR-431-5p/RAB9A axis in melanoma progression. Tumor growth was reduced by ISL in vivo through downregulating circ_0002860 to regulate miR-431-5p and RAB9A levels. Conclusion: The current data indicates that ISL suppressed cell malignant progression of melanoma via targeting the circ_0002860/miR-431-5p/RAB9A pathway.(AU)

Lesions of the oral cavity of dogs: 720 cases / Lesões da cavidade oral de cães: 720 casos

Seven hundred twenty cases of oral cavity lesions in dogs were diagnosed from 2015 to 2020 in Distrito Federal, Brazil. Four hundred thirty-two (60%) lesions were neoplastic, and 288 (40%) were non-neoplastic. Of the neoplasms, 229 (53%) were considered malignant and 203 (47%) benign, affecting mainly males, of 43 different breeds, with an average age of 9.4 years, mainly the gum and lip. Melanoma was the most (96/432, 22.2%) commonly diagnosed neoplasm, followed by peripheral odontogenic fibroma (68/432, 15.7%), papilloma (63/432, 14.5%) and squamous cell carcinoma (SCC) (56/432, 12.9%). Non-neoplastic lesions also affected males predominantly, from 41 different breeds, with an average age of 8.9 years, mainly in the gums, lip, and tongue. These lesions were diagnosed as inflammatory (230/288, 79.8%), hyperplastic (47/288, 16.3%) and other lesions (11/288, 3.8%).

Foram estudados 720 casos de lesões na cavidade oral em cães diagnosticados entre 2015 e 2020, no Distrito Federal. Dessas alterações, 432 (60%) foram neoplásicas e 288 (40%) não neoplásicas. Dentre os neoplasmas, 229 (53%) foram consideradas malignos e 203 (47%) benignos, acometendo principalmente machos, de 43 raças diferentes, com idade média 9,4 anos, afetando principalmente gengiva e lábio. Melanoma foi o neoplasma mais comumente diagnosticado (96/432; 22,2%), seguido do fibroma odontogênico periférico (68/432; 15,7%), papiloma (63/432; 14,5%) e carcinoma de células escamosas (56/432; 12,9%). As lesões não neoplásicas também acometeram predominantemente machos, de 41 raças diferentes, com idade média 8,9 anos, afetando principalmente gengiva, lábio e língua. Essas lesões foram diagnosticadas como inflamatórias (230/288; 79,8%), hiperplásicas (47/288; 16,3%) e como outras alterações (11/288; 3,8%).

Melanomas em cães no Sertão do Nordeste do Brasil - epidemiologia, fatores de risco e achados clinicopatológicos / Melanoma in dogs in the backlands of Northeastern Brazil - epidemiology, risk factors and clinicopathological findings

Background: Melanoma is a malignant neoplasm that arises from melanocytes and malanoblasts. It is also more frequently reported in dogs than in other species. They may arise from melanocytes in the skin, on the surfaces of the mucous membranes, and eyes. The aim of this study was to describe the epidemiological aspects, risk factors and clinicopathological findings of melanoma in dogs in the backlands, northeastern Brazil. Materials, Methods & Results: A retrospective study was carried out in all biopsy samples and necropsy examinations of dogs, from January 2003 to December 2021, at the Animal Pathology Laboratory of the Federal University of Campina Grande, Patos, Paraiba, northeastern Brazil. Epidemiological data, clinical signs, and gross lesions were reviewed from the diagnostic laboratory reports. Samples of the skin, lymph nodes, central nervous system and organs of the thoracic and abdominal cavities were fixed in 10% buffered formalin, processed routinely for histopathology, embedded in paraffin wax, cut into 4 µm sections, and stained with hematoxylin and eosin (HE). Histological sections were also submitted to immunohistochemistry with the primary antibody anti-Melan A. Of the 4717 records found, 1158 (24.5%) were diagnosed with neoplasms, of which 48 (4.14%) cases were of melanoma. Of this total, 28 (58.3%) dogs were elderly, 19 (39.6%) were adults, and 1 (2.1%) was young. Mixed breed animals were the most affected (42.6%), followed by the pinscher breed (19.1%). According to the anatomical region, the most affected site was the skin (38/53=71.7%), followed by the oral cavity (12/53=22.65%) and the eyes (3/53=5.7%). Grossly, the skin lesions were characterized by exophytic and usually blackened, sometimes irregular and firm, nodules. At cut, they had a smooth, compact and blackened surface. Lesions in the oral cavity were characterized by blackened, irregular and infiltrating nodules or masses. The ocular lesions were always unilateral and were characterized by an enlarged and diffusely blackened eyeball, with areas of ulceration and subversion of tissue architecture. In 5 animals there was more than one anatomical site affected, totaling 53 lesions. In 9 (17%) cases, metastases were identified, 8 in regional lymph nodes and 1 in the lung. Histopathology showed a densely non-encapsulated, poorly delimited, expansive and infiltrative neoplasm, composed of neoplastic cells arranged in islands or nests and supported by fibrovascular stroma, containing a variable amount of brownish pigment (melanin). Immunohistochemistry showed strong immunostaining of the neoplastic cells in brown by the anti-Melan A antibody. Discussion: The diagnosis of melanoma was established based on epidemiological, clinical, anatomopathological, and immunohistochemical findings. Gender is not a predisposing factor, and although there was no statistically significant relationship, males were more affected. Senescence is a conditioning risk factor. Elderly animals were more affected (P < 0.0001) than adult ones, with OR = 4.38; and young ones (P = 0.0051), with OR = 12.65. Some breeds, especially those with marked skin pigmentation, were more affected, however the most affected ones in this survey were pinscher and poodle. Cutaneous melanoma accounted for almost 72% of cases, contesting recent studies where oral cavity melanoma was more frequent. Therefore, it is believed that the climatic conditions of the backlands sub-region, in northeastern Brazil, associated with the individual characteristics of the dogs, are involved in the development of these neoplasms, since the climate is predominantly dry, with high temperatures throughout the year, with maximums that can reach 40ºC, favoring the exposure to high incidence of ultraviolet radiation.

Primary perianal malignant melanoma in a dog - combination therapy

Background: Melanocytic neoplasm can arise from melanocytes in any location of the body. Malignant melanoma (MM)has a poor prognosis in dogs and presence of lymphvascular invasion, distant metastasis, or mitotic activity present prognostic value. Primary melanoma affecting the gastrointestinal tract has been rarely reported in veterinary literature, thusthe prognosis affecting gastrointestinal tract is unknown. Electrochemotherapy (ECT) is an effective local treatment whichcombines chemotherapeutic drugs mainly bleomycin or cisplatin followed by the delivery of permeabilizing electricalpulses However, other hydrophilic drugs seem to present an increase cytotoxic effect such as carboplatin.Case: A 9-year-old mixed-breed neutered dog was referred to a private clinic with a mass in the perianal region diagnosedas perianal melanoma. No metastasis was observed on abdominal ultrasound nor chest x-ray (3 views). Clinical signs notedwere tenesmus, hemorrhagic discharge, weight loss and hyporexia. Considering the tumor volume (16.0 x 10.0 cm), a neoadjuvant ECT session was proposed. The authors opted for carboplatin (300 mg/m2, intravenously), administered over 20min and cisplatin intratumorally (1 mg/cm3, equivalent to 1 mL/1cm3 total volume 20 mL) administered in the upper partsof the mass that could be reached while avoiding drug leakage. After administration, sequences of eight biphasic pulses,(100 microseconds), with a voltage ranging from 650-1,000V/cm (pulse generator Onkodisruptor®) using a hexagonal/single pair and plate electrode were delivered. At day 30th, a partial response was observed accordingly to RECIST system,with tumor size of 5.0 x 5.0 cm (65.4 cm3). A second ECT session was performed with the same previous protocol, butwith a decreased dosage of carboplatin (240 mg/m2 consistent with 20% reduction) due to adverse effects in the first session, resulting in stable disease at day 60th (30 days after second ECT). Then...

Primary perianal malignant melanoma in a dog - combination therapy

Background: Melanocytic neoplasm can arise from melanocytes in any location of the body. Malignant melanoma (MM)has a poor prognosis in dogs and presence of lymphvascular invasion, distant metastasis, or mitotic activity present prognostic value. Primary melanoma affecting the gastrointestinal tract has been rarely reported in veterinary literature, thusthe prognosis affecting gastrointestinal tract is unknown. Electrochemotherapy (ECT) is an effective local treatment whichcombines chemotherapeutic drugs mainly bleomycin or cisplatin followed by the delivery of permeabilizing electricalpulses However, other hydrophilic drugs seem to present an increase cytotoxic effect such as carboplatin.Case: A 9-year-old mixed-breed neutered dog was referred to a private clinic with a mass in the perianal region diagnosedas perianal melanoma. No metastasis was observed on abdominal ultrasound nor chest x-ray (3 views). Clinical signs notedwere tenesmus, hemorrhagic discharge, weight loss and hyporexia. Considering the tumor volume (16.0 x 10.0 cm), a neoadjuvant ECT session was proposed. The authors opted for carboplatin (300 mg/m2, intravenously), administered over 20min and cisplatin intratumorally (1 mg/cm3, equivalent to 1 mL/1cm3 total volume 20 mL) administered in the upper partsof the mass that could be reached while avoiding drug leakage. After administration, sequences of eight biphasic pulses,(100 microseconds), with a voltage ranging from 650-1,000V/cm (pulse generator Onkodisruptor®) using a hexagonal/single pair and plate electrode were delivered. At day 30th, a partial response was observed accordingly to RECIST system,with tumor size of 5.0 x 5.0 cm (65.4 cm3). A second ECT session was performed with the same previous protocol, butwith a decreased dosage of carboplatin (240 mg/m2 consistent with 20% reduction) due to adverse effects in the first session, resulting in stable disease at day 60th (30 days after second ECT). Then...(AU)

Rare primary choroidal melanoma in a dog, with optic nerve infiltration / Raro melanoma primário de coroide em cão, com infiltração no nervo óptico

Intraocular melanocytic neoplasms are common in dogs. However, the choroidal location of these neoplasms and their infiltration into the optic nerve are unusual. This paper reports a case of primary choroidal melanoma in a 12-year-old male Dachshund. On the first examination, the animal presented left eye blepharospasm, protrusion of the nictitating membrane, conjunctival hyperemia, moderate congestion, hyphema, and intraocular hypertension. Opacity hindered the performance of fundoscopy in the affected eye. The ocular ultrasound examination revealed a cupuliform mass in the choroid. Due to suspected intraocular neoplasia, an exenteration was performed. Histopathological examination revealed primary choroidal melanoma with scleral and optic nerve invasion.(AU)

Neoplasias melanocíticas intraoculares são comuns em cães, contudo, a localização destas neoplasias na coroide e a infiltração delas no nervo óptico são pouco frequentes. O presente trabalho relata um caso de melanoma de coroide primário em um cão da raça Dachshund, com 12 anos de idade. O animal ao primeiro exame apresentou blefaropasmo no olho esquerdo, protrusão da terceira pálpebra, hiperemia conjuntival, congestão moderada, hifema e pressão intraocular elevada. Opacidade de meios transparentes impediu a realização de fundoscopia no olho acometido. O exame ultrassonográfico ocular revelou massa cupuliforme na coroide. Em razão da suspeita de neoplasia intraocular foi realizada exenteração. No exame histopatológico foi diagnosticado melanoma primário em coroide com invasão escleral e do nervo óptico.(AU)

Retrospective study of cutaneous neoplasms in horses treated at the Livestock Development Center of the Federal University of Bahia / Estudo retrospectivo das neoplasias cutâneas em equinos atendidos no Centro de Desenvolvimento da Pecuária da Universidade Federal da Bahia

The aim of this study was to determine the prevalence of cutaneous neoplasms in horses treated at the Center for the Development of Livestock at the Federal University of Bahia, as well as to correlate it with the coat color, breed, and age of the animal. For that, the attendance records for the last ten years were reviewed. When evaluating the files, 13 cases of cutaneous tumor in horses confirmed by histopathology and cytology were observed. The most prevalent skin tumors were sarcoid (38.5%), melanoma (23%), and fibrosarcoma (15.4%). Regarding the equine coat color, gray and sorrel horses were the most frequent with 30.7% and 23.1% of cases, respectively. As for the equine breed, the mangalarga marchador was the most prevalent (38.4%). Regarding age, 38.46% of the horses were up to 5 years old, 30.77% of the animals were between 4 and 10 years old, and 30.76% were between 11 and 16 years old. In the end, it can be concluded that sarcoid and melanoma were the most prevalent neoplasms.

Objetivou-se com este trabalho determinar a prevalência de neoplasias cutâneas em equinos atendidos no Centro de Desenvolvimento da Pecuária da Universidade Federal da Bahia, bem como correlaciona-la com a pelagem, raça e idade do animal. Para tanto revisou-se as fichas de atendimento dos últimos dez anos. Ao avaliar as fichas, observou-se 13 casos de tumor cutâneo em equinos confirmado por histopatologia ou citologia. Os tumores cutâneos mais prevalentes foram sarcoide (38,5%), melanoma (23%) e fibrossarcoma (15,4%). Com relação a pelagem, equinos tordilhos e alazões foram os mais frequentes com 30,7% e 23,1% dos casos, respectivamente. Quanto as raças, a mangalarga marchador foi a mais prevalente (38,4%). Em relação a idade, 38,46% dos equinos possuíam até 5 anos de idade, 30,77% dos animais apresentavam idade entre 4 e 10 anos e, 30,76% apresentavam idade entre 11 e 16 anos. Ao fim, pode-se concluir que o sarcoide e o melanoma foram as neoplasias mais prevalentes.

Cancer chemopreventive and antioxidant activities of seed, skin, and pulp of maximo hybrid grapes (IAC 138-22) at five different ripening stages / Atividades quimiopreventiva de câncer e antioxidante de sementes, casca e polpa de uva híbridas maximo (IAC 138-22) em cinco diferentes estágios de maturação

This study evaluated the composition and the cancer chemopreventive and antioxidant activities of the seed, skin, and pulp of Maximo hybrid grapes (IAC 138-22) harvested at different ripening stages. The content of total phenolic compounds and tannins were determined by the Folin-Ciocaulteau method, and total anthocyanins were determined using the differential pH method. Different chemical methods assessed the antioxidant activity. Cell viability (macrophage and melanoma skin cancer) and cancer chemopreventive activity were tested in cell lines by quinone reductase induction assay, TNF-α-induced NF-κB inhibitory activity assay, and aromatase inhibitory activity assay. The hybrid grape had high phenolic contents in both seeds and skin, significantly reduced upon ripening. The results showed that fruit ripening significantly affected both the composition and the antioxidant and cancer chemopreventive activities. Except for the anthocyanin content, the most significant results were observed for green seeds, with the greenest stage showing the best results. The green seeds showed the highest antioxidant activity, even when compared to free resveratrol. In the assessment of cancer chemopreventive activity, the green seeds showed the ability to inhibit N-FκB and aromatase activity, important factors for the early stage of cancer development. Besides, no extracts showed cytotoxicity. Therefore, this grape's green seeds showed strong antioxidant and cancer chemopreventive activities confirmed herein, suggesting beneficial health effects.

RESUMO: O objetivo deste estudo foi avaliar a composição e as atividades quimiopreventiva de câncer e antioxidante da semente, polpa e casca de uvas híbridas Maximo (IAC 138-22) colhidas em diferentes estágios de maturação. Os teores de compostos fenólicos totais, antocianinas e taninos foram determinados a partir dos extratos etanólicos. O conteúdo de fenólicos e taninos totais foi determinado pelo método de Folin-Ciocaulteau, e o teor de antocianinas totais foi determinado pelo método do pH diferencial. A atividade antioxidante foi avaliada por diferentes métodos químicos, e a viabilidade celular (macrófagos e melanomas), e atividade quimiopreventiva de câncer foi testada em culturas de células por ensaio de indução de quinona redutase, ensaio de atividade inibidora de NF-κB induzido por TNF-α e ensaio de atividade inibidora de aromatase. A uva híbrida apresentou alto teor de fenólicos nas sementes e na casca, teor que reduziu significativamente com o amadurecimento. Os resultados mostraram que o amadurecimento dos frutos afetou significativamente a composição e as atividades antioxidantes e quimiopreventivas de câncer. Com exceção do teor de antocianinas, os resultados mais significativos foram observados para as sementes verdes, com o estádio mais verde apresentando os melhores resultados. As sementes verdes apresentaram a maior atividade antioxidante, mesmo quando comparadas ao resveratrol livre. Na avaliação da atividade quimiopreventiva do câncer, as sementes verdes mostraram capacidade de inibir a atividade do NF-κB e da aromatase, fatores importantes para o estágio inicial do desenvolvimento do câncer. Além disso, nenhum extrato apresentou atividade citotóxica. Portanto, as sementes verdes desta uva apresentam resultados promissores dadas as fortes atividades antioxidantes e quimiopreventivas do câncer aqui confirmadas, sugerindo efeitos benéficos à saúde.

Pólipo fibroepitelial atípico em córnea de um cão Lhasa Apso / Atypical fibroepithelial polyp in the cornea of a Lhasa Apso dog

Background: Fibroepithelial polyps are benign tumors of mesenchymal and ectodermal origin. Regarding the localization related to the cutaneous region, they may affect the eyelid and periocular area. In veterinary medicine, polyps located in the urethra of dogs and monkeys, vagina of bitches and on the skin have been described. Fibroepithelial polyps may present as single to multiple nodules, exophytic to pedunculated, usually arising on a smooth surface of a common base. The aim of this paper is to describe the ocular and histopathological changes of a corneal fibroepithelial polyp of recurrent character in a dog. Case: A 20-year-old Lhasa Apso dog, with a history of progressive corneal neoformation in the right eye for 4 months. Patient was submitted to routine ophthalmic evaluation. At biomicroscopy, a discrete mucopurulent discharge was observed in the nasal corner, a nodule in the lower eyelid (± 3 mm), moderate conjunctival hyperemia, vascularization and corneal edema between 12 and 5 h, presence of a pinkish proliferative mass in 3 h, affecting the anterior stroma and opacity of the lens, Schirmer's Tear Test (STT) 20 mm/min and intraocular pressure (IOP) 10 mm/Hg. In the left eye, the STT and IOP were within normal range (17 mm/min and 11 mg/Hg, respectively) and at biomicroscopy only lens opacity was observed. Given the presence of the neoformation in the cornea, the patient underwent lamellar keratectomy to excise the mass and third eyelid flap. The tissue was then submitted to histopathology. The histopathological examination showed a proliferation of fibrous component with areas presenting reactive fibroblasts, with acidophilic cytoplasm and with little volume and slight neovascularization, presence of hyperplasia of the lining epithelium and cells with round to oval nuclei, with inconspicuous nucleoli, compatible with fibroepithelial polyp, without evidence of malignancy. After 18 months, the patient presented recurrence of the condition with a history of slow evolution (about ten months). The ophthalmic evaluation showed lesions very similar to the initial one, describing a recurrence of the lesion. Discussion: Corneal neoplasms are uncommon in dogs. The clinical findings in patients with ocular neoplasms include irregular to nodular masses, gray to reddish in color, and typically associated with a vascular supply. These features are related to the type of tumor and its degree of evolution. There are reports in the literature of ocular neoplasms such as squamous cell carcinoma, papilloma, melanocytoma, fibrosarcoma, hemangiosarcoma, melanoma, lymphoma. Related to fibroepithelial polyp with ocular involvement, there are rare reports in human literature and none in veterinary medicine. There is a human description of the presence of a polyp affecting the cornea associated with a corneal dermoid and e other cases of polyps of conjunctival origin. As far as treatment is concerned, the polyp should be surgically removed because of the possibility of recurrence, tissue metaplasia, or even transformation into a malignant tumor, as seen in the case reported here, in which even with surgical removal there was recurrence and metaplasia of the cells. Based on the histopathological findings, it can be concluded that this is a fibroepithelial polyp with a recidivating characteristic, not well described in the veterinary literature, with an atypical presentation affecting the cornea.

Solid iridociliary carcinoma in a dog

Background: Iridociliary epithelial tumors (ICETs) originate from the iris epithelium or ciliary body. They comprise ciliary body adenoma, carcinoma, pleomorphic adenocarcinomas, medulloepitheliomas, and other primitive neuroectodermal tumors. They are the second most common primary intraocular tumors in dogs and have already been reported in sheep and humans. In dogs, they occur more frequently in middle-aged to elderly animals, and the Labrador and Golden Retriever seem to be more predisposed breeds. This study aimed to describe the clinical and pathological aspects of solid iridociliary carcinoma in a dog. Case: A 3-year-old Poodle bitch was treated for discomfort in the left eyeball region, increased intraocular pressure and moderate buphthalmia. A direct ophthalmological examination was performed without equipment, and a mass was visualized in the posterior chamber, distorting the pupillary cleft. We opted for unilateral enucleation and forwarded the material for histological analysis. Macroscopically, the eyeball measured 3.4 cm (anteroposterior) x 2.6 cm (vertical), with a brownish mass that occupied the entire anterior chamber and part of the posterior chamber. Histologically, there was a neoformation in the ciliary body and iris pigment epithelium, partially well-delimited and densely cellular. The neoplasm was organized into predominantly solid formations interspersed with a discrete amount of blood vessels, rare bundles of fibrous stroma, and amorphous eosinophilic material forming membranes that were positive for PAS. Sections of the neoplasm were subjected to immunohistochemistry using anti-cytokeratin AE1/AE3, anti-S100 protein, anti-vimentin, and anti-Ki-67. Positive cytoplasmic immunostaining for cytokeratin and S-100 was observed. Only 45.6% of cells were positive for Ki-67 (500 cells). No immunostaining was observed for vimentin. Discussion: The diagnosis of solid iridociliary carcinoma was based on the histological features and positive immunostaining for cytokeratin AE1/AE3 and protein S100. Iridociliary carcinomas present positive immunostaining for cytokeratin, whereas adenomas and normal iridociliary epithelium do not present this immunostaining. Moreover, the high rate of cell proliferation was indicative of malignant neoplasia, as observed by the high mitotic count and high positivity for Ki-67. The S100 protein helped in the diagnosis of ICETs, as the iridociliary epithelium showed positive staining for this protein. Some histological features are important to consider in the diagnosis of iridociliary tumors in dogs, such as noninvasive growth in the posterior chamber, pigment epithelium, and thick homogeneous membranes on the cell surface. Furthermore, the presence of positive PAS membranes favors the diagnosis of iridociliary epithelial tumors. ICETs must be differentiated from melanocytomas, anterior uveal melanoma, medulloepitheliomas, and metastatic and pleomorphic carcinomas. The histological characteristics, especially the presence of PAS-positive membranes, associated with the immunohistochemical profile of neoplasm cells, help differentiate the ICETs from these tumors. In general, the prognosis is poor for eyeball and vision maintenance in canine iridociliary tumors, and scleral invasion is associated with a higher recurrence rate.

Caryocar villosum attenuates inflammation by inhibiting CXCL1 activation and peripheral hyperalgesia through opioid pathway modulation / Caryocar villosum atenua a inflamação ao inibir a ativação de CXCL1 e a hiperalgesia periférica por meio da modulação da via opioide

The oil of Caryocar villosum is used in Amazonian folk medicine to treat pain and inflammatory conditions. So, we assessed the anti-inflammatory and antinociceptive properties of the ethanolic extract obtained from the fruit peels of this species. The acetic acid-induced writhing, carrageenan-induced mechanical hyperalgesia, formalin, carrageenan-induced paw edema and carrageenan-induced peritonitis tests were used on mice. The C. villosum ethanolic extract significantly inhibited the number of abdominal writhes, mechanical hyperalgesia and paw licking time in the second phase of the formalin test. At a dose of 300 mg kg-1, the extract also significantly reduced the volume of edema formed in the late phase and reduced the recruitment of leukocytes and neutrophils in the peritoneal cavity, as well as CXCL1 chemokine levels. It is suggested that the extract attenuates the leukocyte recruitment by inhibiting the CXCL1 activation. The peripheral antinociceptive activity occured through opioid pathway modulation because pretreatment with C. villosum ethanolic extract reversed the naltrexone-induced antinociception.(AU)

O óleo de Caryocar villosum é usado na medicina popular amazônica para tratar dores e condições inflamatórias. Assim, avaliamos as propriedades antiinflamatórias e antinociceptivas do extrato etanólico obtido das cascas dos frutos desta espécie. Os testes de contorções induzidas por ácido acético, hiperalgesia mecânica induzida por carragenina, formalina, edema de pata induzido por carragenina e peritonite induzida por carragenina foram usados em camundongos. O extrato etanólico de C. villosum obtido das cascas dos frutos inibiu significativamente o número de contorções abdominais, a hiperalgesia mecânica e o tempo de lambida da pata na segunda fase do teste de formalina. Na dose de 300 mg kg-1, o extrato também reduziu significativamente o volume de edema formado na fase tardia e reduziu o recrutamento de leucócitos e neutrófilos na cavidade peritoneal, bem como os níveis de quimiocina CXCL1. Sugere-se que o extrato atenua o recrutamento de leucócitos por meio da inibição da ativação de CXCL1. A atividade antinociceptiva periférica ocorre por meio da modulação da via opioide pois o pré-tratamento com o extrato etanólico de C. villosum reverteu a antinocicepção induzida pela naltrexona.(AU)

Metastatic melanoma in a Saanen goat: clinical, ultrasonographic and anatomopathological aspects - case report / [Melanoma metastático em cabra Saanen: aspectos clínicos, ultrassonográficos e Anatomopatológicos - relato de caso]

This report describes clinical, ultrasonographic and anatomopathological findings in a case of metastatic melanoma in an adult Saanen goat. Clinically, the goat had apathy, an intra-abdominal palpable firm structure, and exophytic keratinized areas on the skin of the udder. Ultrasound revealed non-encapsulated oval structures, with heterogeneous echogenicity and marked central and peripheral vascularization, and hypoechoic hepatic multifocal to coalescent areas. In the udder, there were non-encapsulated oval structures with heterogeneous echogenicity and hyperechoic center surrounded by hypoechogenic tissue. Grossly, there were black multifocal to coalescent areas in the liver, as well as black nodules in mammary and mesenteric lymph nodes, uterus, spleen, and myocardium. Microscopically, multifocal melanocytic neoplastic proliferation was observed in the dermis and junction of the udder epidermis. Most of the neoplastic cells had cytoplasmic granules of melanin. In the liver there were areas of neoplastic tissue compressing the adjacent parenchyma, with central foci of necrosis, mild desmoplasia, and multifocal infiltration of malignant cells into the adjacent tissues. Similar findings were observed in the mammary and mesenteric lymph nodes, uterus, spleen, and myocardium, which characterized metastatic melanoma. Ultrasonography played a key role for establishing the diagnosis of a metastatic melanoma and helped establish a proper clinical management protocol.(AU)

Este relato descreve os achados clínicos, ultrassonográficos e anatomopatológicos em um caso de melanoma metastático em uma cabra Saanen adulta. Clinicamente, a cabra apresentava apatia, estrutura firme palpável intra-abdominal e áreas exofíticas queratinizadas na pele do úbere. A ultrassonografia revelou estruturas ovais não encapsuladas, com ecogenicidade heterogênea e marcada vascularização central e periférica, além de áreas hepáticas multifocais a coalescentes hipoecoicas. No úbere, havia estruturas ovais não encapsuladas, com ecogenicidade heterogênea e centro hiperecogênico circundado por tecido hipoecogênico. Macroscopicamente, havia áreas pretas multifocais a coalescentes no fígado, bem como nódulos pretos nos linfonodos mamários e mesentéricos, no útero, no baço e no miocárdio; microscopicamente, proliferação neoplásica melanocítica multifocal foi observada na derme e na junção da epiderme do úbere. A maioria das células neoplásicas apresentava grânulos citoplasmáticos de melanina. No fígado, havia áreas de tecido neoplásico comprimindo o parênquima adjacente, com focos centrais de necrose, desmoplasia leve e infiltração multifocal de células malignas nos tecidos adjacentes. Achados semelhantes foram observados nos nódulos linfáticos mamários e mesentéricos, no útero, no baço e no miocárdio, que caracterizaram o melanoma metastático. A ultrassonografia desempenhou um papel fundamental para estipular o diagnóstico de um melanoma metastático e ajudou a estabelecer um protocolo de manejo clínico adequado.(AU)

Diffuse melanosis secondary to metastatic melanoma in a Nelore bull

Relatamos um caso de melanose difusa secundária a melanoma maligno metastático em um touro Nelore. Os sinais clínicos incluíram isolamento do rebanho, epistaxe, hipertermia, pálidas membranas oculares, diarreia mucóide e urina escura. Apesar da terapia antiinflamatória e antibiótica, o touro morreu 45 dias após o início dos sinais clínicos. A lesão mais marcante foi uma descoloração negra difusa para os órgãos viscerais, incluindo fígado, baço, pulmões, nódulos linfáticos e rins; todos esses órgãos afetivos foram moderadamente aumentados ”. A urina estava preta. Histologicamente, 50-80% do parênquima do fígado, baço e linfonodos foi obliterado por agregados de melanócitos neoplásicos carregados de melanina. Essas células neoplásicas também ocorreram nos capilares do fígado, baço, linfonodos, bexiga urinária, pulmões e rins. A imunohistoquímica de melanócitos neoplásicos foi positiva para os marcadores Melan A e PNL2. Pigmento marrom a preto abundante foi encontrado em melanófagos nos pulmões, confirmado por imunohistoquímica com IBA1.

Diffuse melanosis secondary to metastatic melanoma in a Nelore bull

Relatamos um caso de melanose difusa secundária a melanoma maligno metastático em um touro Nelore. Os sinais clínicos incluíram isolamento do rebanho, epistaxe, hipertermia, pálidas membranas oculares, diarreia mucóide e urina escura. Apesar da terapia antiinflamatória e antibiótica, o touro morreu 45 dias após o início dos sinais clínicos. A lesão mais marcante foi uma descoloração negra difusa para os órgãos viscerais, incluindo fígado, baço, pulmões, nódulos linfáticos e rins; todos esses órgãos afetivos foram moderadamente aumentados ”. A urina estava preta. Histologicamente, 50-80% do parênquima do fígado, baço e linfonodos foi obliterado por agregados de melanócitos neoplásicos carregados de melanina. Essas células neoplásicas também ocorreram nos capilares do fígado, baço, linfonodos, bexiga urinária, pulmões e rins. A imunohistoquímica de melanócitos neoplásicos foi positiva para os marcadores Melan A e PNL2. Pigmento marrom a preto abundante foi encontrado em melanófagos nos pulmões, confirmado por imunohistoquímica com IBA1.(AU)

Treatment of canine oral melanoma with adjuvant chemotherapy and immunotherapy

Background: Melanoma is the most frequent cancer in the canine oral cavity. It shows an aggressive behavior, characterized by rapid and invasive growth and high metastatic potential. Metastasis is seen in more than 80% of dogs at time ofdeath. Adjuvant therapy should be recommended because of potential recurrence and metastasis. Oral melanoma has a poorprognosis even when adjuvant treatments are used. There are some treatment options, but the high death rate due to thedisease is still a challenge. The aim of this study was to assess the overall survival of dogs diagnosed with oral melanomaand treated with adjuvant chemotherapy and immunotherapy.Materials, Methods & Results: A retrospective analysis was carried out in 20 dogs with oral melanocytic or amelanocyticmelanomas. Cases were staged according to a modified World Health Organization clinical staging system for canine oralmalignant melanoma. Tumor size (T1: 4 cm), regional metastasis (N0: no metastasis; N1:metastasis) and presence of distant metastasis (M0: no metastasis; M1: metastasis) are evaluated. Then, cases were dividedinto 4 stages: I (T1 N0 M0), II (T2 N0 M0), III (T3 N0-1 M0, Tx N1 M0) and IV (Tx Nx M1). Diagnoses were confirmedwith histopathological exam and immunohistochemistry (IHC) when necessary. In poorly differentiated neoplasms, IHC wasperformed at the request of the submitting veterinarian using specific markers PNL-2 and Melan-A. Animals were dividedinto 2 groups: dogs submitted to surgery alone were included in group 1 (G1); dogs submitted...

Treatment of canine oral melanoma with adjuvant chemotherapy and immunotherapy

Background: Melanoma is the most frequent cancer in the canine oral cavity. It shows an aggressive behavior, characterized by rapid and invasive growth and high metastatic potential. Metastasis is seen in more than 80% of dogs at time ofdeath. Adjuvant therapy should be recommended because of potential recurrence and metastasis. Oral melanoma has a poorprognosis even when adjuvant treatments are used. There are some treatment options, but the high death rate due to thedisease is still a challenge. The aim of this study was to assess the overall survival of dogs diagnosed with oral melanomaand treated with adjuvant chemotherapy and immunotherapy.Materials, Methods & Results: A retrospective analysis was carried out in 20 dogs with oral melanocytic or amelanocyticmelanomas. Cases were staged according to a modified World Health Organization clinical staging system for canine oralmalignant melanoma. Tumor size (T1: < 2 cm; T2: 2 - 4 cm; T3: > 4 cm), regional metastasis (N0: no metastasis; N1:metastasis) and presence of distant metastasis (M0: no metastasis; M1: metastasis) are evaluated. Then, cases were dividedinto 4 stages: I (T1 N0 M0), II (T2 N0 M0), III (T3 N0-1 M0, Tx N1 M0) and IV (Tx Nx M1). Diagnoses were confirmedwith histopathological exam and immunohistochemistry (IHC) when necessary. In poorly differentiated neoplasms, IHC wasperformed at the request of the submitting veterinarian using specific markers PNL-2 and Melan-A. Animals were dividedinto 2 groups: dogs submitted to surgery alone were included in group 1 (G1); dogs submitted...(AU)

Isobolographic analysis reveals antinociceptive synergism between Ph1 recombinant toxin and morphine in a model of cancer pain in C57BL/6J mice

Abstract Background: Phoneutria nigriventer venom contains Ph1. This toxin and its recombinant form have a remarkable analgesic potential that is associated with blockage of voltage-gated calcium channels and TRPA1 receptors. Although morphine is a mainstay drug to treat moderate and severe pain related to cancer, it has serious and dose-limiting side effects. Combining recombinant Ph1 and morphine to treat pain is an interesting approach that has been gaining attention. Therefore, a quantitative and reliable method to establish the strength of the antinociceptive interaction between these two substances is necessary. The present study was designed to investigate the nature of the functional antinociceptive (analgesic) interaction between Ph1 recombinant toxin and morphine in a model of cancer pain. Methods: Melanoma was produced by intraplantar inoculation of B16-F10 cells into the right paw of C57BL/6J mice. Von Frey filaments measured the paw-withdrawal threshold after intrathecal administration of morphine, recombinant Ph1, and their combination. Thermal hyperalgesia was assessed using Hargreaves apparatus. The degree of interaction was evaluated using isobolographic analysis. Spontaneous and forced motor performance was assessed with the open-field and rotarod tests, respectively. Results: Co-administration of recombinant Ph1 and morphine synergistically reverses the melanoma-induced mechanical hyperalgesia. The potency of the mixture, measured as the effective dose to reach 50% of maximum possible effect (MPE) in ameliorating mechanical hyperalgesia, was about twice fold higher than expected if the interaction between morphine and recombinant Ph1 was merely additive. Treatment with the combination at doses necessary to reach 50% of MPE caused no spontaneous nor forced motor alterations. Conclusion: The combinatorial use of recombinant Ph1 and morphine allows significant and effective dose reduction of both agents, which has translational potential for opioid-sparing approaches in pain management related to cancer.

Antioxidant, genotoxic, antigenotoxic, and antineoplastic activities of apitoxin produced by Apis mellifera in Northeast, Brazil / Atividades antioxidantes, genotóxicas, antigenotóxicas e antineoplásicas da apitoxina produzida por Apis mellifera no Nordeste do Brasil

The objective was to evaluate the in vitro antioxidant, genotoxic, antigenotoxic, and antineoplastic activities of apitoxin produced by the bee Apis mellifera. The antioxidant activity of the apitoxin solution was evaluated using the DPPH (2,2-diphenyl-1-picrilhydrazyl) method. Genotoxic potential of apitoxin was analyzed by comparing the mean DNA damage indices (idDNA) of L929 strain fibroblasts exposed to hydrogen peroxide (H2O2 - genotoxic substance), distilled water, or apitoxin. The antigenotoxic effect of apitoxin was analyzed by assessing the percentage decrease in H2O2-induced genotoxicity in L929 fibroblasts co-treated with three concentrations of the aqueous apitoxin solution and subjected to comet assay. In vitro antineoplastic activity in human tumor cell lines of prostate adenocarcinoma (PC3), hepatocellular carcinoma (HEPGE2), melanoma (MAD-MB435), and astrocytoma (SNB19), were verified by MTT [3- (4) bromide colorimetric method, 5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium]. Apitoxin had no genotoxic effect on L929 cells at concentrations of 30, 10, and 5 µg/mL after 24 hours of exposure. This effect was only evident at 50 µg/mL. Apitoxin promoted a significant reduction in DNA damage index (idDNA) at all concentrations tested. At 30 µg/mL, apitoxin attenuated the genotoxic effects induced by H2O2. Apitoxin also demonstrated in vitro antineoplastic potential, since the cytotoxic effect was observed at concentrations of 50 µg/mL and 25 µg/mL, with significant reduction in viability percentage of PC3 tumor cell lines, HEPGE2, MAD-MB435, and SNB19. The high antioxidant activity associated with the absence of genotoxic effect and the genoprotective and antineoplastic effect demonstrated by apitoxin here provide indications of apitoxins therapeutic potential.(AU)

O objetivo deste estudo foi avaliar as atividades antioxidantes, genotóxicas, antigenotóxicas e antineoplásicas in vitro da apitoxina produzida pela abelha Apis mellifera. A atividade antioxidante da solução da apitoxina foi avaliada pelo método DPPH (2,2-difenil-1-picrilhidrazil). O potencial genotóxico da apitoxina foi analisado através dos índices médios de dano ao DNA (idDNA) dos fibroblastos da linhagem L929 expostos à peróxido de hidrogênio (H2O2 - substância genotóxica), água destilada ou apitoxina. O efeito antigenotóxico da apitoxina foi analisado através da avaliação da diminuição percentual na genotoxicidade induzida por H2O2 nos fibroblastos L929 co-tratados com três concentrações da solução aquosa de apitoxina e submetidos ao ensaio cometa. A atividade antineoplásica in vitro em linhagens celulares tumorais humanas de adenocarcinoma da próstata (PC3), carcinoma hepatocelular (HEPGE2), melanoma (MAD-MB435) e astrocitoma (SNB19), foram verificadas pelo método colorimétrico do brometo de MTT [3- (4), 5-dimetiltiazol -2-il) -2,5-difeniltetrazólio]. A apitoxina não teve efeito genotóxico nas células L929 nas concentrações de 30, 10 e 5 µg / mL após 24 horas de exposição. Este efeito foi apenas evidente a 50 µg / mL. A apitoxina promoveu uma redução significativa no índice de danos ao DNA (idDNA) em todas as concentrações testadas. A 30 µg / mL, a apitoxina atenuou os efeitos genotóxicos induzidos por H2O2. A apitoxina também demonstrou potencial antineoplásico in vitro, uma vez que o efeito citotóxico foi observado em concentrações de 50 µg / mL e 25 µg / mL, com redução significativa na porcentagem de viabilidade das linhagens celulares de PC3, HEPGE2, MAD-MB435 e SNB19. A alta atividade antioxidante associada à ausência de efeito genotóxico e o efeito genoprotetor e antineoplásico demonstrado pela apitoxina aqui fornecem indicações do potencial terapêutico da apitoxina.(AU)

Isobolographic analysis reveals antinociceptive synergism between Phα1ß recombinant toxin and morphine in a model of cancer pain in C57BL/6J mice

Background: Phoneutria nigriventer venom contains Phα1ß. This toxin and its recombinant form have a remarkable analgesic potential that is associated with blockage of voltage-gated calcium channels and TRPA1 receptors. Although morphine is a mainstay drug to treat moderate and severe pain related to cancer, it has serious and dose-limiting side effects. Combining recombinant Phα1ß and morphine to treat pain is an interesting approach that has been gaining attention. Therefore, a quantitative and reliable method to establish the strength of the antinociceptive interaction between these two substances is necessary. The present study was designed to investigate the nature of the functional antinociceptive (analgesic) interaction between Phα1ß recombinant toxin and morphine in a model of cancer pain. Methods: Melanoma was produced by intraplantar inoculation of B16-F10 cells into the right paw of C57BL/6J mice. Von Frey filaments measured the paw-withdrawal threshold after intrathecal administration of morphine, recombinant Phα1ß, and their combination. Thermal hyperalgesia was assessed using Hargreaves apparatus. The degree of interaction was evaluated using isobolographic analysis. Spontaneous and forced motor performance was assessed with the open-field and rotarod tests, respectively. Results: Co-administration of recombinant Phα1ß and morphine synergistically reverses the melanoma-induced mechanical hyperalgesia. The potency of the mixture, measured as the effective dose to reach 50% of maximum possible effect (MPE) in ameliorating mechanical hyperalgesia, was about twice fold higher than expected if the interaction between morphine and recombinant Phα1ß was merely additive. Treatment with the combination at doses necessary to reach 50% of MPE caused no spontaneous nor forced motor alterations. Conclusion: The combinatorial use of recombinant Phα1ß and morphine allows significant and effective dose reduction of both agents, which has translational potential for opioid-sparing approaches in pain management related to cancer.(AU)

Melanose talâmica em caprinos / Thalamic melanosis in goats

Background: Melanosis is a blackened pigmentation resulting from the accumulation of melanocytes in tissues that are not normally pigmented. This change in the color of the organs occurs due to the agglomeration of melanocytes originating from abnormal migration during embryogenesis and does not cause dysfunction to the affected organ. Although melanosis frequently occurs in several species and affects several organs such as the brain and spinal cord leptomeninges, involvement in the thalamus region is unusual. The objective of this work was to report 2 cases of thalamic melanosis in goats, determining the pathological and histochemical aspects that assist in the diagnosis of this condition. Cases: Two cases of thalamic melanosis in goats were diagnosed. In both cases, the animals had no nervous history disease and clinical signs. The cause of death in cases 1 and 2 was established based on anatomopathological findings and clinical signs being diagnosed with mycoplasmosis and asphyxia, respectively. After fixing and making cross-sections of the brain, a focal lengthy blackened area was observed on the thalamus surface in both cases. Microscopically, lesions in the brain were similar in both cases and exclusively affected the thalamus. These cells had abundant cytoplasm, well delimited with brownish granular pigment. The nuclei were difficult to visualize and in some cells, it was rounded, well-defined, morphologically compatible with melanocytes. Melanocytes were mainly distributed around neurons and often distended the perivascular space of multiple blood vessels. In Fontana Masson staining, the granules in the cytoplasm of these cells stained strongly black. The Prussian Blue, Periodic Acid- Schiff's, Von Kossa, and Giemsa stains were negative, and the pigment remained brown. In the unstained slides, assembled after the deparaffinization and clarification process, it was observed the permanence of cells with blackish-brown pigment in the cytoplasm. In immunohistochemistry, strong immunostaining of pigmented cells with the Anti-MelanA antibodies was observed in both cases. Discussion: The diagnosis of thalamic melanosis in goats was carried out based on the characteristic pathological findings, in which melanin pigments were demonstrated and identified through HE, Fontana-Masson staining, and unstained slides and confirmed by the IHC. The use of complementary histochemical techniques was fundamental for the classification of the pigment as melanin, demonstrating to be an accessible and reliable tool for the diagnosis of pathological processes that lead to the accumulation of pigments and or material in the tissues. The occurrence of melanin in the thalamus may be associated with a failure in the migration of melanoblasts, which would go to the optical pathways or to the thalamus. This erratic migration of melanoblasts can be explained by the fact that the forebrain is the embryogenic origin of the optic and diencephalon pathways. Macroscopically, thalamic melanosis must be differentiated mainly from neoplastic processes such as melanoma and hemangiosarcoma, pigmented fungus infections, Phalaris angusta poisoning, listeriosis, neurocutaneous melanosis, and neuromelanin. It was concluded that thalamic melanosis is an uncommon alteration in goats and although it has been diagnosed as an incidental necropsy finding, should be included in the differential diagnosis of diseases that affect the central nervous system, especially those that have a color change associated with the deposition of pigments in the tissues.