Clinical and molecular aspects of breast cancer: Targets and therapies.

Breast Cancer is a complex disease characterized by the occurrence of multiple molecular alterations. Currently, some molecular markers are in use for breast cancer diagnostic, prognostic, and predictive purposes. Thus, genetic signatures are available for improving the decision-making. The biomarkers are also essential as therapeutic approaches, but many questions remain due to the lack of efficacy on breast cancer treatment, mainly for triple-negative breast cancer subtype. Since the genetic profile of breast cancer can also be related to different ethnic groups and geographic areas, the reference populations of the genetic assays and clinical trials need to include a broader population beyond the European and North American patients. In this review, we analyzed the current and potential molecular markers that could help to improve the strategies for breast cancer therapy.

Parotid gland adenocarcinoma in a cow: case report / Adenocarcinoma de glândula parótida em vaca: relato de caso

A 9-year-old Girolando dairy cow, weighing 400kg, with a history of increased volume in the right parotid region, which extended to the submandibular region, was assisted. Fine needle aspiration cytology was performed, and the cytological findings were consistent with malignant neoplasm of epithelial origin (carcinoma). Because of the unfavorable prognosis, the animal was euthanized and submitted to an anatomopathological examination. Samples of the increased parotid and affected lymph nodes were collected for histopathological evaluation. The microscopic changes were accentuated features of anaplasia, moderate cell proliferation, atypical mitotic figures, and necrosis. Stroma ranged from delicate to scirrhous, and the tumor boundaries were not distinct. These findings substantiated the preliminary histomorphological diagnosis of undifferentiated carcinoma with metastasis in lymph nodes. Immunohistochemical tests were performed with anti-CK Pan (clone AE1AE3), anti-CK HMW (clone 34ßE12), anti-CK19 (clone RCK108), anti-vimentin (clone V9), anti-S100 (polyclonal), and anti-androgen (polyclonal) antibodies. The immunophenotype favored the diagnosis of salivary gland adenocarcinoma. Despite the rareness in cattle, salivary gland adenocarcinoma should be considered in the differential diagnosis of diseases that occur with increased volume in the head, lymphadenopathy, drooling, dysphagia, and progressive weight loss.(AU)

Foi atendida uma vaca da raça Girolando, de nove anos de idade, de aptidão leiteira, pesando aproximadamente 400kg e com histórico de aumento de volume na região parotídea e submandibular direita. Diante do prognóstico desfavorável, o animal foi submetido à eutanásia e encaminhado para exame anatomopatológico. Fragmentos da glândula parótida e dos linfonodos alterados foram colhidos e encaminhados para exame histopatológico. À avaliação microscópica, observaram-se acentuada anaplasia, moderada proliferação celular, figuras de mitose atípicas e focos de necrose. O estroma variava de delicado a esquirroso e os limites do tumor eram imprecisos. Esses achados fundamentaram o diagnóstico de carcinoma indiferenciado com metástase em linfonodos. No exame imuno-histoquímico, foram utilizados anticorpos primários monoclonais anti-CK Pan (clone AE1AE3), anti-CK alto peso molecular (clone 34ßE12), anti-CK19 (clone RCK108), antivimentina (clone V9), anti-S100 (policlonal) e antirreceptor de andrógenos (policlonal). As células neoplásicas apresentaram imunomarcação para todos os anticorpos testados, resultado que favorece o diagnóstico de adenocarcinoma de glândula salivar. Embora raro em bovinos, o adenocarcinoma de glândula salivar deve ser considerado no diagnóstico diferencial de doenças que cursam com aumento de volume na cabeça, linfadenopatia salivação, disfagia e emagrecimento progressivo.(AU)

Fine needle aspiration cytology in feline skeletal muscle as a diagnostic tool for extramedullary plasmacytoma / Citologia aspirativa por agulha fina no músculo esquelético felino como ferramenta diagnóstica para o plasmocitoma extramedular

Extramedullary noncutaneous plasmacytoma (ENP) is a myeloproliferative disorder of plasma cells that rarely affects cats. This paper describes an ENP case revealed by fine needle aspiration cytology (FNAC) of the mass in the skeletal muscle of an 8-month-old, male, mixed breed cat, which had a nodule in the left hind limb. The rapid immunoassay test confirmed the presence of feline leukemia virus (FeLV). The animal necropsy macroscopically showed the nodule came from the semimembranosus muscle. Histopathological examination ratified the cytological findings. Thus, this paper alerts to the existence of plasmacytoma located in the skeletal muscle of feline species. FNAC is a quick and efficient method for diagnosis of ENP.(AU)

O plasmocitoma extramedular (PEM) não cutâneo é um distúrbio mieloproliferativo de plasmócitos que raramente acomete felinos. Este trabalho descreve um caso de PEM no músculo esquelético de um gato, macho, sem raça definida, de oito meses de idade, que apresentava um aumento de volume no membro pélvico esquerdo. A citologia aspirativa por agulha fina (CAAF) da massa revelou tratar-se de PEM. O teste imunoenzimático rápido confirmou a presença do vírus da leucemia felina (FeLV). Na necropsia do animal, macroscopicamente, percebeu-se que o nódulo era originário do músculo semimembranoso. O exame histopatológico ratificou os achados citológicos. Desta forma, alerta-se para a existência de plasmocitoma com localização em músculo esquelético na espécie felina, sendo a CAAF um método alternativo rápido e eficaz para o seu diagnóstico.(AU)

Metabolism of broilers subjected to different lairage times at the abattoir and its relationship with broiler meat quality / Metabolismo de frangos de corte submetidos a diferentes tempos de espera no abatedouro e sua relação com a qualidade da carne

An investigation was made into the effects of different lairage times and the position of chicken crates during transport to the slaughterhouse on the biochemical and hematological profile and physical parameters of broilers, such as color and pH of their breast meat. The treatments were defined by the animals slaughtered after 0, 2, 4 and 6 hours of lairage time at the slaughterhouse, transported in crates located in the top and bottom layers of the truck. It was found that increasing the lairage time at the slaughterhouse to over two hours reduced the number of lymphocytes and increased the heterophil/lymphocyte (H/L) ratio and the basophil count in the hemogram. In addition, lactate dehydrogenase (LDH) activity and cholesterol levels increased and plasma triglyceride and glucose levels decreased. The position of the crates in the truck altered the creatine kinase (CK) activity, and the highest enzyme activity was found in birds transported in the top layer of crates in the truck. Furthermore, the long lairage time in the slaughterhouse increased the pH and the value of a* (redness value) and decreased the lightness value of breast fillets. The interaction significant between 4 and 6 hours of lairage time and the position of the crate in the top layer of the truck favored the development of dark, firm, dry (DFD) meat.(AU)

Este trabalho objetivou a avaliação dos efeitos dos diferentes tempos de espera e da posição das caixas de transporte no caminhão sobre perfil bioquímico e hematológico, além de parâmetros físicos da carne, como cor e pH do peito, de frangos de corte. Os tratamentos foram definidos pelos animais abatidos com zero, duas, quatro e seis horas de espera no abatedouro, posicionados nas partes superior e inferior do caminhão. Como resultado, verificou-se que o aumento do tempo de espera no abatedouro, acima de duas horas, resultou em redução no número de linfócitos, elevação da razão heterófilos/ linfócitos (H/L) e de basófilos no hemograma. Houve aumento da atividade de lactato desidrogenase (LDH), dos níveis de colesterol e redução de triglicerídeos e glicose no plasma. O posicionamento das caixas na parte superior da carroceria do caminhão elevou a enzima creatina quinase (CK) sanguínea. Além disso, o tempo prolongado na área de espera aumentou o pH final e o valor de a* (teor de vermelho) e diminuiu a luminosidade dos filés. A interação significante dos fatores tempo de espera de quatro e seis horas e a posição superior das caixas de transporte propiciaram o desenvolvimento de carnes duras, firmes e escuras (DFD) em frangos de corte.(AU)

Molecular analysis and conventional cytology: association between HPV and bacterial vaginosis in the cervical abnormalities of a Brazilian population.

We investigated the association between bacterial vaginosis (BV) and human papillomavirus (HPV) infection in Papanicolaou smears in a Brazilian population. Cross-sectional analysis was performed on 673 samples collected from women attending public health centers in Olinda (PE, Brazil) by conventional cytology methodology and molecular analysis, PCR tests (GP5+/6+ and MY09/11). Cytological abnormalities, BV, and HPV-DNA were detected in 23 (3.4%) samples, 189 samples (28.1%), and 210 samples (31.2%), respectively. GP5+/6+ primers resulted in higher detection performance than MY09/11 primers, with 81% concordance between both primers (P < 0.0001). The occurrence of HPV-DNA and BV had ORs of 8.59 (P < 0.0001) and 2.91 (P = 0.0089) for abnormal cytology, respectively, whereas the concomitant presence of both infections showed an OR equal to 3.82 (P = 0.0054). Therefore, we observed an association between abnormal cervical cytology and HPV infection, BV, or both HPV infection and BV. These results highlight the necessity of monitoring patients presenting not only HPV, but also BV, as risk factors for cervical lesion development.

Células-tronco mononucleares associadas ao plasma rico em plaquetas na consolidação de falha óssea no cão. Estudo piloto / Mononuclear stem cells associated with platelet-rich plasma in the consolidation of bone defects in dogs. A pilot study

A padronização de técnicas e o teste da associação do Plasma Rico em Plaquetas (PRP) e das Células-Tronco Mononucleares (CTMs) na consolidação de falhas ósseas corticais, por meio de avaliação clínica, biomecânica, radiológica e histológica, é avaliada em um estudo piloto. Foram utilizados seis cães adultos, fêmeas, sem raça definida, pesando entre 5 e 10kg, separados por sorteio aleatório em seis tratamentos. Foi confeccionada uma falha elíptica de 1,0x0,4cm na cortical medial diafisária da tíbia direita de cada animal, sendo preenchida de acordo com o tratamento proposto. No cão I, a falha foi preenchida com solução fisiológica (SF); no II, com o PRP; no III, com a fração total das células mononucleares (FTCM); no IV, com a fração vascular estromal (FVE); no V, com o PRP associado à FTCM; no VI, com a associação PRP e FVE. Foram realizadas avaliações: clínicas, diariamente; dos graus de claudicação, semanalmente; radiológica e perimetria da coxa, antes, no pós-operatório imediato, aos 7, 14, 21 e 30 dias; biomecânica, antes do procedimento, aos 10, 20 e 30 dias; e biópsias, aos 15 e 30 dias. A FTCM obteve uma contagem e viabilidade média de 2,0x108cél. e 90%, respectivamente, enquanto a FVE obteve 3x106cél. e 50%. O PRP concentrou, em média, sete vezes o número inicial de plaquetas do sangue total, de 250.000 µl-1 plaquetas no sangue total para 1.750.000 µl-1 plaquetas no PRP. Obteve-se padronização adequada de técnicas, possibilitando o teste da associação entre as células-tronco mononucleares (CTMs) e o plasma rico em plaquetas (PRP), assim como seu uso isolado, no reparo de falhas ósseas corticais, indicando a possibilidade de a associação FTCM e PRP ser o melhor tratamento...

The standardization of techniques and tests of the association of Platelet Rich Plasma (PRP) and Mononuclear Stem Cells (MSCs) in the consolidation of cortical bone defects by clinical, biomechanical, radiological, and histological analysis is evaluated in a pilot study. Six adult female dogs of mixed breed, weighing between 5 and 10kg, separated by random draw in six treatments were used. An elliptical failure of 1.0 x0.4cm was done in the medial diaphyseal cortical of the right tibia of each animal, that was filled according with the proposed treatment. In dog I, the failure was filled with saline (S), in dog II with PRP, in dog III with total mononuclear cell fraction (TMCF), in dog IV with stromal vascular fraction (SVF), in dog V with association of PRP and TMCF, and in dog VI with an association of PRP and SVF. Daily clinical evaluation, weekly degrees of lameness, radiological and girth before, immediate postoperative, 7, 14, 21 and 30 days, biomechanics before the procedure, at 10, 20 and 30 days, and biopsies at 15 and 30 days were performed. The TMCF got a count and viability of 2,0x108cells and 90% respectively, while for SVF it was 3x106cells and 50%, respectively. The PRP concentrated on average seven times the original number of platelets from whole blood, platelets from whole blood 250.000 μl-1 to 1.750.000 μl-1 platelets in PRP. This afforded adequate standardization of techniques, enabling the test of association between mononuclear stem cells (MSCs) and platelet-rich plasma(PRP), as well as their separate use to repair cortical bone defects, indicating the possibility of the association between FTCM and PRP to be the best treatment...

Antioxidant and anti-inflammatory effects of quercetin in functional and morphological alterations in streptozotocin-induced diabetic rats.

The aim of this study was to investigate functional and morphological alterations caused by oxidative stress in streptozotocin (STZ)-induced diabetic rats and to evaluate the antioxidant effect of quercetin (QUE) in this disease. One hundred and thirty male Wistar rats, it were randomly distributed in 10 different experimental groups, with ten animals per group: Control Saline (CS), Control Ethanol (CE), Control QUE 5mg/kg (CQ5), Control QUE 25mg/kg (CQ25), Control QUE 50mg/kg (CQ50), Diabetic Saline (DS), Diabetic Ethanol (DE), Diabetic QUE 5mg/kg (DQ5), Diabetic QUE25 mg/kg (DQ25), Diabetic QUE 50mg/kg (DQ50). Therefore, hyperglycemia is directly involved in oxidative stress production, as well as in functional and morphological alterations caused by the excess of free radicals. QUE, specially at the dosage of 50mg/kg, can act as an antioxidant and anti-inflammatory agent, becoming a promising adjuvant in the treatment of diabetes mellitus.

Peroxidação lipídica em ratos tratados com sulfato de vincristina e decanoato de nandrolona / Lipid peroxidation in rats treated with vincristine sulphate and nandrolone decanoate

Brain and serum lipid peroxidation was studied in rats treated with vincristine sulphate and different doses of nandrolone decanoate. Thirty rats were distributed into six groups (n=5). The treatments were applied once a week for two weeks. Sample collection was performed in the third week. Treatments during the first week were: G1 (control) - physiologic solution, G2 - vincristine sulphate (4mg/m²), G3 - physiologic solution, G4 - physiologic solution, G5- vincristine sulphate (4mg/m²), and G6 - vincristine sulphate (4mg/m²). In the second week, they were: G1 (control) - physiologic solution, G2- physiologic solution, G3 - nandrolone decanoate (1.8mg/kg-1), G4 - nandrolone decanoate (10mg/kg-1), G5 - nandrolone decanoate (1.8mg/kg-1), and G6 - nandrolone decanoate (10mg/kg-1). Lipid peroxidation increased with the isolated use of vincristine and nandrolone decanoate, and with vincristine associated to the highest dose of the ester as well. These results suggest that vincristine sulphate and nandrolone decanoate increase free radical production. Therapeutic dose of nandrolone decanoate when associated with vincristine sulphate proved to be beneficial, as it was able to protect the organism from damaging processes involved in free radical production.

Este estudo teve por objetivo detectar a peroxidação lipídica presente no cérebro e no soro de ratos tratados com sulfato de vincristina e diferentes doses de decanoato de nandrolona. Trinta ratos foram distribuídos em seis grupos (n=5). Os tratamentos foram aplicados uma vez por semana, durante duas semanas, e a coleta de amostras foi realizada na terceira semana. Na primeira semana, os tratamentos consistiram de: G1(controle) - solução fisiológica; G2 - sulfato de vincristina (4mg/m 2 ); G3 - solução fisiológica; G4 - solução fisiológica; G5 - sulfato de vincristina (4mg/m 2 ) e G6 - sulfato de vincristina (4mg/m 2 ). Na segunda semana: G1(controle) - solução fisiológica; G2 - solução fisiológica; G3 - decanoato de nandrolona (1.8mg/kg-1 ); G4 - decanoato de nandrolona (10mg/kg-1 ); G5 - decanoato de nandrolona (1.8mg/kg-1 ) e G6 - decanoato de nandrolona (10mg/kg-1 ). A peroxidação lipídica aumentou com o uso isolado tanto da vincristina quanto do decanoato de nandrolona e com a associação da vincristina à dose mais alta do éster. Estes resultados sugerem que o sulfato de vincristina e o decanoato de nandrolona aumentam a produção de radicais livres. A dose terapêutica do decanoato de nandrolona, quando associada ao sulfato de vincristina, provou ser benéfica, já que foi capaz de proteger o organismo dos processos prejudiciais induzidos pela produção de radicais livres.

Células-tronco mononucleares autólogas e proteína óssea morfogenética na cicatrização de defeitos tibiais experimentalmente induzidos em cães / Autologue mononuclear stem cells and morphogenetic bone protein in experimentally induced tibial defect healing in dogs

Avaliou-se a utilização de células-tronco mononucleares (CTM) na cicatrização de defeito ósseo experimental como alternativa aos métodos convencionais, analisando-se o tempo de evolução cicatricial e a presença dessas células no tecido neoformado. Foram utilizados 18 cães, separados em três grupos (G) de seis, e de cada animal foram colhidas células da medula óssea (MO), contadas e analisadas para morfometria, por meio da contagem manual e mielograma. Um defeito ósseo tibial foi então criado cirurgicamente, e a lesão tratada com esponja de gelatina embebida em solução fisiológica (G1), esponja de gelatina embebida com aspirado de MO processado (G2) e esponja de gelatina embebida com aspirado de MO processado e proteína óssea morfogenética (rhBMP-2) (G3). A cicatrização foi então avaliada por estudos radiográficos, e a presença de CTM foi identificada por meio de marcadores nanocristais Qtracker, em microscopia com luz fluorescente, uma semana após a intervenção cirúrgica. Entre as células identificadas pelo marcador, foram encontradas células da linhagem óssea. As avaliações radiográficas demonstram crescimento ósseo acelerado nos animais de G2 e G3. Houve diferenças significativas entre o G1 e G3 em todos os tempos estudados, e entre G1 e G2 nos tempos de 30 e 45 dias. A utilização de CTM adultas suplementadas ou não com rhBMP-2 é alternativa favorável ao crescimento ósseo em defeitos experimentais agudos de tíbia de cães.

Mononuclear stem cells (MSC) were experimentally implanted in bone defect, as an alternative to the conventional methods, in order to evaluate the healing speed, and the presence of these cells in the new-born tissue. Bone marrow (BM) was collected from 18 dogs, and then counted and morphometrically analyzed by manual count and myelogram. The dogs were separated in three groups (G) of six animals each. A tibial bone defect was surgically made in each dog and the wound was treated with gelatin sponge and physiologic solution (G1), gelatin sponge and processed BM (G2), and gelatin sponge, processed BM, and rhBMP-2 (G3). The healing was evaluated by radiographic study and the presence of MSC was microscopically identified by Qtracker nanocrystal labeler with a fluorescent light one week after the surgery. Cells from the bone lineage were found among the labeled cells. The radiographic evaluations demonstrated a speed up in bone growth in dogs from G2 and G3, and significant differences were found between G1 and G3 dogs, in all studied periods; and between G1 and G2 animals at 30 and 45 days. The rhBMP-2 supplemented or not-supplemented adult MSC are favorable alternatives to the bone growth in the healing process of acute experimental-induced tibial defects in dogs.

HPV31 and HPV33 incidence in cervical samples from women in Recife, Brazil.

Human papillomavirus (HPV) has been extensively studied concerning genomic structure, infection mechanisms, and diversity of types, as well as disease progression stages and development of vaccines. HPV type prevalence can differ in specific populations in different countries, according to ethnicity. This is the first report of an integrated project to evaluate the incidence of HPV types in different regions in Brazil in order to obtain data for vaccine development. Cervical samples were collected from women seen at a public hospital in Pernambuco, Northeast Brazil, for routine evaluation of genital alterations. Selection of the patients was random. There was a strong prevalence of HPV16 and a high incidence of HPV types 31 and 33. These data foster the discussion about the need to evaluate viral prevalence in each geographic region in order to develop targeted vaccine programs.

Implantação do programa de fitoterapia "Verde Vida" na secretaria de saúde de Maringá (2000-2003)

Há muitos anos, tenta-se implantar a fitoterapia no município de Maringá, o que só foi possível em 2000, com a criação de uma comissão específica, definição das plantas segundo critérios epidemiológicos, literatura disponível e adoção do formato de farmácia de manipulação. Foram padronizados os fitoterápicos nas formas de infusão (espinheira-santa, guaco, cavalinha, hortelã e funcho), gel (própolis e calêndula) e em cápsulas gelatinosas duras (valeriana). Encontrou-se problemas com fornecedores, com diversos lotes insatisfatórios quanto à qualidade, mas ao longo do tempo o programa foi muito bem aceito. Nestes três anos de funcionamento, já foram dispensados mais de 50.000 unidades de produtos, com destaque à aceitação e eficácia da valeriana como ansiolitico, dos géis de própolis e calendula como antimicrobiano e cicatrizante, da espinheirasanta em gastrites e úlceras e do guaco como broncodilator e expectorante. Confirma-se, assim, o valor da fitoterapia no apoio às atividades básicas de saúde.

There are many years, it tries to implant the phytotherapy in the municipal district of Maringá. That implantation was only possible in 2000, with the creation of specific commission, definition of the plants according to epidemic criteria and available literature, and adoption of the format of manipulation drugstore. We have standardized the phytotherapics in the infusion forms (espinheira-santa, guaco, cavalinha, mint and fennel), gel (própolis and calendula) and in hard gelatinous capsules (valerian). There were problems with suppliers, with several unsatisfactory lots as for the quality, but along the time the program was very well accepted. On these three years of operation, there were already released more than 50.000 units of products, with prominence to the acceptance and effectiveness of the valerian as ansiolitic, two gels of propolis and calendula as antimicrobial and healing, of the espinheirasanta in gastritis and ulcers and of the guaco as broncodilator and expectorant. It is confirmed, like this, the value of the phytotherapy in the support to the basic activities of health.