Effects of the juçara fruit (Euterpe edulis Martius) pulp and lyophilized extract on NRF2, KEAP1, SOD1, and GPX2 expression in human colorectal cancer cell lines.

We investigated the effects of the juçara fruit (Euterpe edulis Martius) pulp and lyophilized extract on the expression of cytoprotective genes nuclear factor erythroid 2 (NF-E2)-related factor 2 (NRF2), kelch-like ECH-associated protein 1 (KEAP1), superoxide dismutase (SOD1), and glutathione peroxidase (GPX2) in human colorectal cancer cell lines (HT-29 and Caco-2). Cells were cultured for 24 h in Dulbecco's Modified Eagle's Medium containing juçara fruit pulp (5, 10, or 50 mg/mL) or lyophilized extract (0.05, 0.1, or 0.5 mg/mL), and gene expression was quantified using real-time quantitative reverse transcription polymerase chain reaction. All studied genes showed significant variation in gene expression among different concentrations of pulp or lyophilized extract. Overall, the expression of the selected genes decreased in both cell lines following exposure to the pulp or lyophilized extract in a dose-dependent manner for most of the concentrations studied. In summary, our study showed that the compounds in juçara fruit inhibited the expression of cytoprotective genes associated with the antioxidant response and that, although not cytotoxic at the concentrations studied, they could potentially block the activation of the NRF2/KEAP1 pathway.

Effects of the juçara fruit (Euterpe edulis Martius) pulp and lyophilized extract on NRF2, KEAP1, SOD1, and GPX2 expression in human colorectal cancer cell lines

We investigated the effects of the juçara fruit (Euterpe edulis Martius) pulp and lyophilized extract on the expression of cytoprotective genes nuclear factor erythroid 2 (NF-E2)-related factor 2 (NRF2), kelch-like ECH-associated protein 1 (KEAP1), superoxide dismutase (SOD1), and glutathione peroxidase (GPX2) in human colorectal cancer cell lines (HT-29 and Caco-2). Cells were cultured for 24 h in Dulbecco's Modified Eagle's Medium containing juçara fruit pulp (5, 10, or 50 mg/mL) or lyophilized extract (0.05, 0.1, or 0.5 mg/mL), and gene expression was quantified using real-time quantitative reverse transcription polymerase chain reaction. All studied genes showed significant variation in gene expression among different concentrations of pulp or lyophilized extract. Overall, the expression of the selected genes decreased in both cell lines following exposure to the pulp or lyophilized extract in a dose-dependent manner for most of the concentrations studied. In summary, our study showed that the compounds in juçara fruit inhibited the expression of cytoprotective genes associated with the antioxidant response and that, although not cytotoxic at the concentrations studied, they could potentially block the activation of the NRF2/KEAP1 pathway.

Aberrant right subclavian artery in a dog - case report / [Ectopia de artéria subclávia direita em cão - relato de caso]

The brachiocephalic trunk and the left subclavian artery originate from the aortic arch, and both supply blood to the head, neck, and thoracic limbs. Anatomical variations, such as an aberrant right subclavian artery, are congenital conditions rarely observed in dogs, Thus, the objective of the present report was to describe a case of aberrant right subclavian artery in a 9-year-old Dalmatian. However, this anomaly was a finding in which the patient was asymptomatic during its 9 years of life and only at this age did he exhibit signs including sialorrhea, vomiting, hyporexia, and noisy deglutition. Blood count, biochemical profile, and thoracic radiography led to a diagnosis of megaesophagus and aspiration pneumonia. Despite the recommended treatment, the patient did not respond well; as such, the owner elected to euthanize the animal. On necropsy, the right subclavian artery originated directly from the aortic arch, followed a route from left to right dorsally to the esophagus, and then formed an impression of the vascular path over the muscular wall of the esophagus. The esophagus, in turn, exhibited a flaccid wall and dilation in the caudal portion to the vascular path made by the ectopic position of the right subclavian artery.(AU)

O tronco braquiocefálico e a artéria subclávia esquerda emergem do arco aórtico e são responsáveis por fazerem o suprimento sanguíneo para cabeça, pescoço e membros torácicos. Variações anatômicas, como a ectopia da artéria subclávia direita, são alterações congênitas raramente encontradas em cães, cujas alterações do sistema digestivo acontecem em pacientes recém-desmamados e não em adultos. Assim, o objetivo deste relato é descrever um caso de ectopia da artéria subclávia direita em uma cadela, Dálmata, de nove anos de idade. No entanto, essa anomalia foi um achado do qual o paciente foi assintomático durante os nove anos de vida e somente com essa idade apresentou sinais como sialorreia, vômito, hiporexia e deglutição ruidosa. O hemograma e os perfis bioquímicos, associados à radiografia torácica, levaram a um diagnóstico de megaesôfago e pneumonia aspirativa. Mesmo seguindo o tratamento recomendado, houve piora clínica do quadro e o animal foi submetido à eutanásia. À macroscopia, a artéria subclávia direita originava-se direto do arco aórtico, fazia um percurso da esquerda para a direita dorsalmente ao esôfago e, então, formava uma impressão do trajeto vascular sobre a parede muscular do esôfago. O esôfago, por sua vez, apresentava parede flácida e dilatação na porção caudal ao trajeto vascular feito pela posição ectópica da artéria subclávia direita.(AU)

Melanose multicêntrica em frango de corte: relato de caso / Multicentric melanosis in broiler: case report

Melanoblastos podem migrar de forma errática durante a embriogênese, gerando um quadro conhecido como melanose. São raros os estudos envolvendo melanose com acometimento múltiplo dos órgãos. Objetivou-se descrever um caso de melanose multicêntrica em frango de corte que gerou condenação do animal ao abate. Foram encaminhadas ao Laboratório de Patologia Animal da Universidade Federal do Espírito Santo para avaliação histopatológica amostras de diversos órgãos de uma ave de corte da linhagem Cobb de 48-49 dias de idade. Esse animal foi condenado na linha de inspeção por apresentar áreas multifocais enegrecidas. Ao exame microscópico, observou-se melanina multifocal variando de moderada a intensa quantidade em todos os órgãos acometidos com lesões enegrecidas na macroscopia. As áreas pigmentadas foram negativas para a coloração especial de azul da Prússia e não foram encontradas células neoplásicas. A pigmentação da pele em aves comercializadas no Ocidente é rara, pois a característica de pele clara foi selecionada por meio de melhoramento genético, mas pouco se sabe sobre quais mutações desencadeiam melanose nas linhagens selecionadas para não apresentar pigmentação. A ave apresentou melanose multicêntrica e tal alteração não compromete a função dos órgãos acometidos nem representa risco para a saúde humana, no entanto, devido ao aspecto macroscópico, os órgãos que apresentam tal lesão foram condenados para consumo humano.(AU)

Melanoblasts can enter an erratic migratory pathway during embryogenesis and it creates a clinical condition known as melanosis. Studies involving melanosis in multiple organs are rare. The aim of this paper is to describe a case of multicentric melanosis in a broiler resulting in animal condemnation at slaughter. Samples from diverse organs originating from a Cobb broiler, 48-49 days of age, were sent to Laboratório de Patologia Animal from Universidade Federal do Espírito Santo for histopathological avaliation. This animal was condemned in line inspection due to multifocal black areas of pigmentation. At microcopic avaliation, multifocal melanin in moderate-intense amount in all the organs stricken with macroscopic black lesions was noted. Sections of pigmented areas were negative for Prussian Blue and no neoplasic cells were found. Pigmentation of the skin in a broiler from the Ocident is rare because the white/yellow skin was selected through genetic enhancement, but there is some data about which mutations unleash melanosis in lineages selected to not have pigmentation. The broiler had multicentric melanosis and this alteration doesn't change functional activity of the stricken organs and doesn't represent any risk for human health, but the organs with this lesion were condemned for human consumption due to their macroscopic appearance.(AU)

Multiple congenital defects in a newborn foal / Defeitos congênitos múltiplos em um potro recém-nascido

A case of multiple congenital defects in a newborn foal is reported. The animal showed hypoplasia of the left pelvic limb bones, uterus unicornis, congenital diaphragmatic hernia, and unilateral renal and ureteral agenesis. This report includes the macroscopic and microscopic lesions observed in the case...

Relata-se um caso de defeitos congênitos múltiplos em um potro recém-nascido. O animal apresentou hipoplasia dos ossos do membro pélvico esquerdo, útero unicorno, hérnia diafragmática congênita e agenesia renal e ureteral unilateral. Este relato inclui as lesões macroscópicas e microscópicas observadas no caso...

Diffuse heterotopic hair in a canine uterus: case report / Heterotopia de pelo difusa no útero de uma cadela: relato de caso

The present report describes the clinical and pathological findings related to diffuse heterotopic hairs associated with pseudo-placentational endometrial hyperplasia in a canine uterus. Macroscopically, the uterus was intensely enlarged and thicker, and the endometrial surface was irregular and covered with black hairs similar to the hairs of the bitch. Histologically, heterotopic hairs associated with pseudo-placentational endometrial hyperplasia were observed.

O presente relato descreve os achados clínicos e patológicos de uma heterotopia de pelos difusa associada com hiperplasia endometrial pseudo-placentacional em um útero de uma cadela. Macroscopicamente, o útero estava intensamente aumentado de volume e espesso, a superfície do endométrio estava irregular e recoberta com pelos pretos semelhantes aos pelos da cadela. Histologicamente, heterotopia de pelos associada com hiperplasia endometrial pseudoplacentacional foi observada.

Leiomioma ovariano, tumor de células da granulosa e ovário policístico em cadela: uma rara associação / Ovarian leiomyoma, granulosa cell tumor and polycystic ovary in a bitch: a rare association

This case describes a rare case of ovarian leiomyoma associated with granulosa cell tumor and polycystic ovary and in a fifteen-year-old mongrel female dog.

Primary ovarian rhabdomyosarcoma in a dog.

A 10-year-old female English pointer was diagnosed with an ovarian tumour with abdominal metastases. Ultrasonography revealed several nodules of 1-5 cm diameter within the abdominal cavity. Fine needle aspiration cytology of the nodules suggested a malignant mesenchymal tumour. On necropsy examination the right ovary and its capsule were enlarged and there were white-red, friable nodular masses distributed over the surface of the pancreas, liver, omentum, mesentery and serosae of the small and large intestines. Microscopical evaluation revealed neoplastic cells with a high degree of pleomorphism and vascular invasion. Immunohistochemically, the neoplastic cells expressed myosin, desmin, vimentin and CD10, but were negative for cytokeratin, placental alkaline phosphatase, inhibin-alpha and smooth muscle actin. Based on these findings a diagnosis of primary ovarian alveolar rhabdomyosarcoma was made.

Protective effect of rPb40 as an adjuvant for chemotherapy in experimental paracoccidioidomycosis.

The conventional treatment for the most prevalent mycosis in Latin America, paracoccidioidomycosis (PCM), involves long periods of therapy that results in side effects and a high frequency of relapses. The search for a new, alternative treatment is necessary. Pb40 is an antigenic protein from P. brasiliensis fraction F0. This fraction has already been shown to have significant protective activity when used as a PCM vaccine in experimental models. The complete cDNA sequence corresponding to Pb40 was cloned into a pET-21a plasmid, expressed in E. coli with a his-tag and purified by affinity chromatography. The predicted protein sequence exhibited nearly 100% homology to a fragment of the hypothetical EF-hand domain containing protein of P. brasiliensis. Immunization with this recombinant protein was used together with chemotherapy in an attempt to improve PCM treatment. The combined drug/rPb40 treatment exhibited long-lasting control of PCM in the liver and spleen and largely preserved the tissue structures of these organs. Despite the lack of a reduction in CFUs in the group that received the combined treatment, there was a significant reduction in the size of the lesions in the lungs after 70 days of infection. At the same time, the IL-10 levels were higher in the treated mice than in the infected-only mice. Moreover, significant levels of rPb40-specific IgG antibodies were detected in the sera of immunized mice. Thus, the treatment protocol consisting of rPb40 immunization in addition to fluconazole chemotherapy showed an additive protective effect after intratracheal challenge, preventing fungal dissemination to other sites of infection and preventing relapses. These results provide new prospects for PCM immunotherapy.

Physical activity improves age-related decline in the osteogenic potential of rats' bone marrow-derived mesenchymal stem cells.

AIM: To examine whether physical activity increases osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs) from adult rats compared with young rats. METHODS: Eighteen female Wistar rats were divided into three groups and the following cells isolated: (1) differentiated BMMSCs from young donors, (2) differentiated BMMSCs from sedentary adult donors and (3) differentiated BMMSCs from active adult donors. We analysed MTT conversion, percentage of cells per field, mineralized nodule number and gene expression for telomerase reverse transcriptase (TERT), alkaline phosphatase, caspase 3, osteocalcin, bone sialoprotein and collagen I. RESULTS: Telomerase reverse transcriptase expression and the percentage of cells per field in BMMSCs cultures from adult rats were smaller than those observed in young donors. However, levels of caspase 3 expression were higher in BMMSCs from adult donors (P < 0.05). Despite the fact that physical activity was associated with an increase in expression of caspase 3 (P < 0.05), there was no difference in the percentage of cells per field between groups of adult BMMSCs (active or sedentary). However, physical activity increased the number of mineralized nodules and osteocalcin expression after 21 days, and alkaline phosphatase expression at 7, 14 and 21 days in the BMMSCs of adult donors (P < 0.05). However, those values were smaller when compared with young donors BMMSCs (P < 0.05). Only the expression levels of alkaline phosphatase were similar to young donors BMMSCs (P ≥ 0.05). CONCLUSION: Physical activity increases osteogenic differentiation of BMMSCs from adult donors but does not increase the differentiation to the levels observed in BMMSCs from young donor rats.

Mielograma de ratas com disfunção tireoidiana / Bone marrow examination in rats with thyroid dysfunction

The cells of the myelo id, lymphoid , and erythroid lineage s of the bone marrow were quantified in rats with hypo and hyperthyroidism. Fifteen Wistar rats were divided into three groups: hypothyroid (n=5), hyperthyroid (n=5) , and control (n=5). Three months after the onset of the treatment s, euthanasia was performed . Bone marrow was aspirated from femurs of each animal to perform smear s that were stained with Quick Panoptic. T he percentage s of rubroblast, prorubrocyte, metarubrocyte, myeloblast, promyelocytes, metamyelocytes, myel ocytes, segmented, eosinophils, basophils, lymphocytes, plasma cells and monocytes in were determined a total of 500 cells. The bone marrow of animals with hypothyroidism had hypoplasia. The myeloid:erythroid ratio was higher in animals with thyroid dysfun ction. In hypo and hyperthyroidism, there was a significant reduction of the percentage of rubrocyte, metarubrocyte , and lymphocytes and increase of myelocytes and segmented cells. In hypothyroidism, there was a significant increase in the percentage of me tamyelocytes. It is c oncluded that both hypo and hyperfunction of thyroid increase the myeloid:erythroid ratio by increasing the number of cells of the myeloid lineage and reducing the cells of the erythroid lineage.

Administration of thyroxine affects the morphometric parameters and VEGF expression in the uterus and placenta and the uterine vascularization but does not affect reproductive parameters in gilts during early gestation.

The aim of this study was to evaluate the effects of thyroxine administration on morphometric parameters, expression of vascular endothelial growth factor (VEGF) and vascularization in the uterus and placenta and reproductive parameters in gilts at 70 days of gestation. At 150 days of age, i.e., before first heat, 20 gilts were randomly divided into two experimental groups: treated (n=10) and control (n=10). The treated group received a daily dose of 400 µg of L-thyroxine (T(4)) in their diet until slaughter and the control group received only typical meals. Before artificial insemination, blood was collected to determine plasma total T(4). The gilts were inseminated in the second oestrus and slaughtered at 70 days of gestation. The foetal thyroid follicular epithelium height, number, size and weight of foetuses; foetal myogenesis, corpora lutea number, embryonic mortality rate, uterine weight, placental weight and placental fluid volume were measured. Histomorphometric and immunohistochemical analysis of uterus and placenta were determined. The averages of all variables were compared by the Student's t-test. The gilts treated with thyroxine showed significant increase of plasma total T(4). At 70 days of gestation, the heights of the trophoblastic epithelium, endometrial epithelium and endometrial gland epithelium were significantly higher in the group treated with T(4). The expression of cytoplasmatic and nuclear VEGF in trophoblastic cells and the number of blood vessels per field in endometrial stroma were significantly higher in the gilts treated with T(4). No other significant differences between groups were obtained with respect to other parameters (p>0.05). We conclude that oral administration of T(4) up to 70 days of pregnancy in gilts affects the morphometric parameters, the expression of placental VEGF and the uterine vascularization but does not affect reproductive parameters in gilts during early gestation.

Dose-dependent effects of triiodothyronine on the osteogenic differentiation of rat bone marrow mesenchymal stem cells.

BACKGROUND/AIMS: The aim of this study was to investigate the dose-dependent effects of triiodothyronine (T3) on the osteogenic differentiation of mesenchymal stem cells(MSCs). METHODS: MSCs that express CD73, CD54 (intercellular adhesion molecule-1) and CD90 were cultured in triplicate (1 x 10(5)/well) in osteogenic medium with T3 (1, 10, 10(3) or 10(5) pM) or without T3 (control) for 7, 14 and 21 days. Alkaline phosphatase activity, conversion of MTT into formazan crystals, collagen synthesis, collagen maturation, the number of mineralized nodules and their diameters were all determined, and the means were compared by the Student-Newman-Keuls test. RESULTS: A dose of 10(5) pM T3 resulted in a negative effect on MSC osteogenic differentiation, with less collagen synthesis. The 1 pM T3 dose resulted in greater collagen synthesis and alkaline phosphatase activity and more mineralized nodules than in the control group, similar to the 10 pM dose. Nevertheless, the 10 pM dose demonstrated better results than the 1 pM dose with regard to MSC osteogenic differentiation, with greater MTT reduction, better collagen maturation and a larger mean diameter of mineralized nodules. CONCLUSIONS: The effect of T3 on MSC differentiation is dose-dependent, with the 10 pM dose promoting better bone marrow MSC osteogenic differentiation.

Association of nucleolar organizing regions and Ki-67 expression with recurrence rate of hair follicle tumor in dogs / Associação das regiões organizadoras de nucléolos e da expressão do Ki-67 com a taxa de recorrência dos tumores de folículo piloso em cães

Mitotic index, nuclear diameter, number of nucleolar organizing regions, and Ki-67 expression, in hair follicle tumors of 82 dogs were evaluated. Tissue specimens were used to prepare sections for histological staining for number of nucleolar organizing region and immunohistochemical staining for Ki-67. Tumors were classified as trichoblastoma (n=32), benign trichoepithelioma (n=30), pilomatricoma (n=7), malignant trichoepithelioma (n=6), infundibular keratinizing acanthoma (n=5), and tricholemmoma (n=2). Head, dorsum, and limbs were the most affected sites. Malignant trichoepithelioma presented significantly higher mitotic index, number of nucleolar organizing regions and Ki-67 expression. Regarding benign neoplasms, trichoblastoma presented significantly higher mitotic index and number of nucleolar organizing regions. Ki-67 expression did not differ among hair follicle benign neoplasms. Recurrence was only observed in two cases, with one benign trichoepithelioma and one malignant trichoepithelioma. In the two cases, nodules have not been removed with clean surgical margin. It was concluded that in benign neoplasms of hair follicles, the number of nucleolar organizing regions and Ki-67 expression were significantly smaller than in malignant neoplasm. Clean surgical margins are suggested to be more responsible to tumor recurrences than the number of nucleolar organizing regions, expression of Ki-67, and the mitotic index.

O objetivo deste trabalho foi avaliar o índice mitótico, o diâmetro nuclear, o número de regiões organizadoras de nucléolos e a expressão do Ki-67 em 82 tumores de folículo piloso de cães, entre 2000 e 2006. Os tumores foram classificados como tricoblastoma (n=32), tricoepitelioma benigno (n=30), tricoepitelioma maligno (n=6), pilomatricoma (n=7), acantoma infundibular ceratinizante (n=5) e tricolemoma (n=2). A cabeça, o dorso e os membros foram os locais mais frequentemente acometidos. O tricoepitelioma maligno apresentou índice mitótico, número de regiões organizadoras de nucléolos e expressão do Ki-67 significativamente maiores quando comparado aos outros tipos de tumores. Dentre as neoplasias benignas, o tricoblastoma apresentou índice mitótico e número de regiões organizadoras de nucléolos significativamente maiores. A expressão do Ki-67 não diferiu entre os tumores benignos de folículo piloso. A recorrência foi observada apenas em dois casos, incluindo um tricoepitelioma benigno e um tricoepitelioma maligno. Em dois casos, os nódulos não foram removidos com margem cirúrgica completa. Desta forma, conclui-se que nas neoplasias benignas de folículo piloso, o número das regiões organizadoras de nucléolo e da expressão de Ki-67 foram significativamente menores que em neoplasias malignas. Além disto, sugere-se que a margem cirúrgica completa está mais associada à recorrência dos tumores que as regiões organizadoras de nucléolo, a expressão de Ki-67 e o índice mitótico.

Osteogenic differentiation of mesenchymal stem cells from osteopenic rats subjected to physical activity with and without nitric oxide synthase inhibition.

Physical activity has potent and complex effects on bones. We hypothesized that physical activity has a positive effect upon osteopenic rat bones because it stimulates osteogenic differentiation of bone marrow mesenchymal stem cells (MSCs). We also postulated that local nitric oxide concentrations mediate the effects of physical activity on bones. The objective of this study was to investigate the osteogenic differentiation in vitro of MSCs from osteopenic female rats subjected to physical activity with and without nitric oxide synthase inhibition. We used MSCs from the femurs of Wistar female rats divided into six groups: Group 1, sham-operated (control); Group 2, sedentary osteopenic; Group 3, active osteopenic; Group 4, sham-operated with L-NAME; Group 5, sedentary osteopenic with L-NAME; and Group 6, active osteopenic with L-NAME. The cells were cultured at 37 degrees C and 5% CO2. Cells were phenotypically characterized with anti-CD45, anti-CD90, anti-CD73, and anti-CD54 using a FACScan cytometer. MSCs were cultured in osteogenic medium for 7, 14 and 21 days. Alkaline phosphatase activity, the capacity of dimethylthiazol conversion in formazan crystals, collagen synthesis and the number of mineralized nodules were analyzed. The means of all of the variables were compared using the SNK test. MSCs did not express CD45 in 96.94% of the cells, but there was expression of CD73, CD54 and CD90 in 93.99%, 95.10% and 86.77% of the cells, respectively. MSCs from osteopenic rats showed less osteogenic differentiation. Surprisingly, physical activity increased the osteogenic differentiation of MSCs in osteopenic rats. Inhibition of nitric oxide synthase in vivo had a negative effect upon the osteogenic potential of MSCs from normal rats and from osteopenic rats subjected to physical activity. Our results suggest that nitric oxide stimulates MSCs osteogenic differentiation and that nitric oxide mediates the beneficial effects of physical activity upon MSCs osteogenic differentiation.