Crude extract of Jatobá leaves promotes canine osteosarcoma cell D17 proliferation.

Background and Aim: New substances for neoplasm treatment have to be carefully studied to minimize adverse effects and prevent disease progression stimulation. Jatobá is a typical tree of the Cerrado and Caatinga biome, with antifungal, antimicrobial, larvicide, antioxidant, and antiproliferative properties. This study aimed to investigate the action of the crude extract of Jatobá leaves (EBFJ) on canine osteosarcoma (CO) cells and analyze the expression of biomarkers in neoplasm progression. Materials and Methods: D17 cells were cultured and subjected to treatment with EBFJ at different concentrations (10 µg/mL; 100 µg/mL; 1000 µg/mL; 2000 µg/mL; and 5000 µg/mL) and exposure times (24 h, 48 h, and 72 h). The tetrazolium reduction assay and the immunocytochemistry technique, with anti-Bcl2, anti-p53, and anti-Ki-67 antibodies, were used to observe the effect of the extract on cell proliferation. Results: Doses of 2000 µg and 5000 µg had cell viability of 300.80% and 361.84%, respectively. The extract did not show significant cytotoxicity of samples with the control group. The confluence of cells, the number of labeled cells, and the expression of Bcl2, Ki-67, and p53 were higher in the groups treated with EBFJ, with a statistical difference from the group without treatment. Conclusion: EBFJ was not cytotoxic and had a proliferative effect on CO D17 cells. The confluence of cells, the number of labeled cells, and the expression of Bcl2, Ki-67, and p53 were higher in the groups treated with the extract.

Oxidonitrergic and antioxidant effects of a low molecular weight peptide fraction from hardened bean (Phaseolus vulgaris) on endothelium.

About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction <3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 µg/mL PV3. Oxidative stress was provoked by 3% H2O2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO- and endothelium-dependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 µg/mL PV3. PV3 at 30 µg/mL increased cell viability, while cytotoxicity was observed only with 250 µg/mL PV3. PV3 at 10 µg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H2O2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release.

Oxidonitrergic and antioxidant effects of a low molecular weight peptide fraction from hardened bean (Phaseolus vulgaris) on endothelium

About 3000 tons of beans are not used in human food due to hardening. Several studies on bean-derived bioactive peptides have shown potential to treat some diseases, including those relying on oxidative dysfunctions. We assessed the effects of peptides extracted from hardened bean Phaseolus vulgaris (PV) on reactive oxygen species (ROS) and nitric oxide (NO) production, cytotoxic and cytoprotective effects in endothelial cells, and oxidonitrergic-dependent vasodilating effects. Extract was composed by peptide fraction <3 kDa (PV3) from hardened common bean residue. PV3 sequences were obtained and analyzed with bioinformatics. Human umbilical vein endothelial cells were treated with 10, 20, 30, and 250 µg/mL PV3. Oxidative stress was provoked by 3% H2O2. Cytotoxicity and cytoprotective effects were evaluated by MTT assay, whereas, ROS and NO were quantified using DHE and DAF-FM fluorescent probes by confocal microscopy. NO- and endothelium-dependent vasodilating effects of PV3 were assessed in isolated aortic rings. We found 35 peptides with an average mass of 1.14 kDa. There were no cell deaths with 10 and 20 μg/mL PV3. PV3 at 30 μg/mL increased cell viability, while cytotoxicity was observed only with 250 μg/mL PV3. PV3 at 10 μg/mL was able to protect cells from oxidative stress. PV3 also increased NO release without causing cell death. It also reduced relative ROS production induced by H2O2. PV3 vasodilating effects relied on endothelium-dependent NO release. PV3 obtained from low-commercial-value bean displays little cytotoxicity and exerts antioxidant effects, whereas it increases endothelial NO release.

Histological classification and expression of markers of canine mast cell tumors.

BACKGROUND AND AIM: Mast cell tumors (MCTs) are malignant neoplasms that are common in dogs. Their biological behavior is variable and unpredictable. The aim of the present study was to analyze the histological classification and expression of markers of canine MCTs. MATERIALS AND METHODS: Thirty samples of canine MCTs were graded according to the histological classification methods of Patnaik and those of Kiupel. The expression of phosphoprotein 53 (p53) and c-kit proteins was quantified by immunohistochemistry using image processing software, ImageJ - a public domain computer program, developed at the National Institutes of Health. RESULTS: It was possible to determine the grade of 100% of the samples. According to Patnaik's classification, 20.00% of the samples were Grade 1, 43.30% were Grade 2, and 36.70% were Grade 3. According to Kiupel's classification, 56.67% of the samples were of high intensity and 43.33% were of low intensity. Grade 1 tumors had the highest expression of p53 and c-kit, and Grade 2 had the lowest expression. The results showed that it is necessary to perform both histological grading methods. The classification into high and low intensity may provide more consistent results than the three-level grading system. However, a smaller number of categories, although it facilitates the classification, may not be sufficient for the prognosis. CONCLUSION: Quantitative evaluation of p-53 and c-kit expression is a useful tool to increase the accuracy of the analysis and to aid in choosing the treatment method for canine MCTs. Histological grading should be combined with other diagnostic methods.

Anatomical aspects of the nerves of the leg and foot of the giant anteater (Myrmecophaga tridactyla, Linnaeus, 1758) / Aspectos anatômicos dos nervos da perna e pé do tamanduá-bandeira (Myrmecophaga tridactyla, Linnaeus, 1758)

Although distal stifle joint nerve distribution has been well established in domestic animals, this approach is scarcely reported in wild animals. Therefore, the aim of this study was to describe the nerves of the leg and foot of Myrmecophaga tridactyla with emphasis on their ramification, distribution, topography and territory of innervation. For this purpose, six adult cadavers fixed and preserved in 10% formalin solution were used. The nerves of the leg and foot of the M. tridactyla were the saphenous nerve (femoral nerve branch), fibular and tibial nerves and lateral sural cutaneous nerve (branches of the sciatic nerve) and caudal sural cutaneous nerve (tibial nerve branch). The saphenous nerve branches to the skin, the craniomedial surface of the leg, the medial surface of the tarsal and metatarsal regions and the dorsomedial surface of the digits I and II (100% of cases), III (50% of cases) and IV (25% of cases). The lateral sural cutaneous nerve innervates the skin of the craniolateral region of the knee and leg. The fibular nerve innervates the flexor and extensor muscles of the tarsal region of the digits and skin of the craniolateral surface of the leg and dorsolateral surface of the foot. The tibial nerve innervates the extensor muscles of the tarsal joint and flexor, adductor and abductor muscles of the digits and the skin of the plantar surface. The caudal sural cutaneous nerve innervates the skin of the caudal surface of the leg. The nerves responsible for the leg and foot innervation were the same as reported in domestic and wild animals, but with some differences, such as the more distal division of the common fibular nerve, the absence of dorsal metatarsal branches of the deep fibular nerve and a greater involvement of the saphenous nerve in the digital innervation with branches to the digits III and IV, in addition to digits I and II...

Apesar de bem estabelecida nos animais domésticos, a abordagem da distribuição nervosa distal do joelho é rara em animais selvagens. Portanto, o objetivo deste estudo foi descrever os nervos da perna e pé do Myrmecophaga tridactyla, com ênfase na sua ramificação, distribuição, topografia e território de inervação. Para tanto, foram utilizados seis cadáveres adultos, fixados e conservados em solução de formalina a 10%. A dissecação envolveu desde a formação dos nervos femoral e isquiático pelos ramos ventrais dos nervos espinhais lombares e sacrais até sua distribuição nos territórios propostos. Os nervos responsáveis pela inervação da perna e pé do M. tridactyla foram o N. safeno (ramo do N. femoral), os nervos fibular comum e tibial e o N. cutâneo lateral da sura (derivados do N. isquiático) e o N. cutâneo caudal da sura (ramo do N. tibial). O nervo safeno emite ramos cutâneos para a superfície craniomedial da perna, medial do tarso e metatarso e dorsomedial dos dedos I e II (100% dos casos), III (50% dos casos) e IV (25% dos casos). O nervo cutâneo lateral da sura inerva a região cutânea craniolateral do joelho e perna. O nervo fibular inerva os músculos flexores do tarso e extensores dos dedos e a região cutânea craniolateral da perna e dorsolateral do pé. O nervo tibial inerva os músculos extensores do tarso e flexores, adutores e abdutores dos dedos e região cutânea plantar. O nervo cutâneo caudal da sura inerva a pele da face caudal da perna. Pode-se concluir que os nervos responsáveis pela inervação da perna e pé foram os mesmos relatados em animais domésticos e selvagens, porém com algumas diferenças, como a divisão mais distal do nervo fibular comum, ausência de ramos metatarsianos dorsais do N. fibular profundo e uma maior participação do nervo safeno na inervação digital, contribuindo com ramos inclusive para os dedos III e IV, além dos dedos I e II...

Gross anatomy of the brachial plexus in the giant Anteater (Myrmecophaga tridactyla).

Ten forelimbs of five Myrmecophaga tridactyla were examined to study the anatomy of the brachial plexus. The brachial plexuses of the M. tridactyla observed in the present study were formed by the ventral rami of the last four cervical spinal nerves, C5 through C8, and the first thoracic spinal nerve, T1. These primary roots joined to form two trunks: a cranial trunk comprising ventral rami from C5-C7 and a caudal trunk receiving ventral rami from C8-T1. The nerves originated from these trunks and their most constant arrangement were as follows: suprascapular (C5-C7), subscapular (C5-C7), cranial pectoral (C5-C8), caudal pectoral (C8-T1), axillary (C5-C7), musculocutaneous (C5-C7), radial (C5-T1), median (C5-T1), ulnar (C5-T1), thoracodorsal (C5-C8), lateral thoracic (C7-T1) and long thoracic (C6-C7). In general, the brachial plexus in the M. tridactyla is similar to the plexuses in mammals, but the number of rami contributing to the formation of each nerve in the M. tridactyla was found to be larger than those of most mammals. This feature may be related to the very distinctive anatomical specializations of the forelimb of the anteaters.

Estudo anatômico do plexo lombossacral de Tamandua tetradactyla / Anatomical study of the lumbosacral plexus of the Tamandua tetradactyla

O tamanduá-mirim (Tamandua tetradactyla) é um xenartro da família Myrmecophagidae, encontrado da Venezuela ao sul do Brasil. Estudos apontam que essa é uma das espécies de animais selvagens mais vitimadas em número de atropelamentos, e, muitas vezes, o atendimento clínico adequado aos indivíduos feridos é dificultado pela carência de informações acerca dos mesmos. Visando contribuir com o conhecimento dessa espécie, este estudo teve como objetivo descrever seu plexo lombossacral. Para tanto, foram utilizados quatro cadáveres de Tamandua tetradactyla adultos e de ambos os sexos. O plexo lombossacral dessa espécie é formado pelos ramos ventrais dos nervos espinhais T18, L1, L2, L3, S1, S2, S3, S4, S5. Os nervos integrantes do plexo lombossacral do T. tetradactyla com suas formações mais frequentes foram os seguintes: genitofemoral (T18), cutâneo femoral lateral (T18-L1), femoral (T18, L1-L3), obturador (T18, L1-L3), glúteo cranial (L3-S1), isquiático (L3-S3), pudendo (S3-S4 ou S4-S5), retal caudal (S4 ou S5) e cutâneo femoral caudal (S4-S5). O plexo lombar e sacral dessa espécie é unido, sendo L3 o ponto de união entre eles. Devido ao pequeno número de vértebras lombares, a composição dos nervos do plexo lombossacral do T. tetradactyla apresenta características peculiares que se diferem das características das demais espécies já estudadas, quais sejam, a ausência dos nervos ílio-hipogástrico e ilioinguinal e participação de nervos torácicos na composição dos nervos do plexo lombar, presença de contribuição sacral na composição do nervo obturador e ausência de contribuição lombar na composição do nervo isquiático e um limite mais caudal na extensão do plexo sacral.

The lesser anteater (Tamandua tetradactyla) is a xenarthra of the Myrmecophagidae family found from Venezuela to southern Brazil. Studies have shown that this is one of the most numerous wildlife species victims of car collisions on roads, and often the appropriate clinical care to injured animals is hindered by the lack of information about them. In order to contribute to the knowledge of this species, this study aimed to describe its lumbosacral plexus. For this purpose, four cadavers of adult specimens of both sexes of T. tetradactyla were used. The lumbosacral plexus of the T. tetradactyla is formed by the ventral rami of spinal nerves T18, L1, L2, L3, S1, S2, S3, S4, S5. The lumbosacral plexus nerves with their most common formations in this species were as follows: genitofemoral (T18), lateral femoral cutaneous (T18-L1), femoral (T18, L1-L3), obturator (T18, L1-L3), cranial gluteal (L3-S1), ischiatic (L3-S3), pudendus (S3-S4 or S4-S5), caudal retal (S4 or S5), and caudal femoral cutaneous (S4-S5). The lumbar and sacral plexus of this species is joined, L3 being the link between them. Due to the small number of lumbar vertebrae, the arrangement of the lumbosacral plexus nerves of the T. tetradactyla showed peculiar characteristics that differ it from that of other previously studied species, such as the absence of iliohypogastric and ilioinguinal nerves and contribution of thoracic nerves in the formation of all the nerves of the lumbar plexus, presence of sacral contribution in the formation of the obturator nerve, and the lack of lumbar contribution for sciatic nerve formation and a most caudal extent of the sacral plexus.