Nuclear proliferation marker MIB-1 (Ki-67) immunohistochemistry (IHC) is used to examine tumor cell proliferation. However, the diagnostic or prognostic value of the Ki-67 nuclear staining intensity and location, defined as nuclear gradient (NG), has not been assessed. This study examined the potential association between Ki-67 NG and cell cycle phases and its effect on the prognosis of pulmonary typical carcinoid (PTC) tumors. We propose a method for classifying the NG of Ki-67 during the cell cycle and compare the results between PTC, pulmonary adenocarcinoma (PAD), and breast ductal carcinoma (BDC). A literature review and objective analysis of IHC-stained paraffin sections were used to determine the Ki-67 labeling index and composed a stratification of the NG into NG1, NG2, and NG3/4 categories. A semi-automated image analysis protocol was established to determine the Ki-67 NG in PTC, PAD, and BDC. High intraobserver consistency and moderate interobserver agreement were achieved in the determination of Ki-67 NG in tumor specimens. NG1 and NG2 were lower in PTC than in PAD and BDC. Cox multivariate analysis of PTC after adjusting for age and number of metastatic lymph nodes showed that Ki-67 NG1 and NG2 significantly predicted clinical outcomes. The semi-automated method for quantification of Ki-67 nuclear immunostaining proposed in this study could become a valuable diagnostic and prognostic tool in PTC. MIB1 MIB 1 MIB- Ki67 Ki 67 (Ki-67 IHC (IHC However Ki-6 location NG, , (NG) assessed (PTC tumors (PAD) BDC . (BDC) IHCstained stained NG34 NG3 4 NG3/ categories semiautomated semi automated specimens outcomes Ki6 6 (Ki-6 Ki- (NG (PAD (BDC (Ki- (Ki

RESUMO O objetivo deste estudo foi verificar a capacidade de diferenciação das células-tronco da polpa dentária canina em células progenitoras neurais bem como quantificar obtenção e viabilidade celular, durante três passagens em cultura. As células foram extraídas da polpa dentária de dois cadáveres caninos, com aproximadamente dez meses de idade, que foram a óbito em decorrência de traumatismo automotivo. Após três subculturas, realizou-se avaliação da viabilidade celular por quantificação em câmara de Neubauer. A partir disso, induziu-se diferenciação neural em meio de cultura neurobasal (Gibco™), com células aderidas ao plástico ou suspensas em placas tratadas com agarose. Após sete e 14 dias em cultivo indutor, observou-se morfologia e perfil imunofenotípico utilizando citometria de fluxo e imunocitoquímica fluorescente. Aos 14 dias as células apresentaram alto grau de expressão para marcadores anti-nestina e anti-glial fibrillary acidic protein (anti-GFAP). Anteriormente, obteve-se ao 25º dia, média de 18x10⁶ células viáveis indiferenciadas oriundas do tecido pulpar. Sugere-se que as células-tronco indiferenciadas da polpa dentária canina apresentem índices satisfatórios de diferenciação em células progenitoras neurais, aderidas ou suspensas em cultura. A polpa dentária dos dentes decíduos caninos, fornece células indiferenciadas viáveis em quantidade adequada.

ABSTRACT The objective of this study was to verify the differentiation capacity of canine tooth pulp stem cells in neural progenitor cells as well as to quantify the attainment and viability during three culture passages. The cells were extracted from the dental pulp of two canine cadavers, with approximately ten months of age, which died due to automotive trauma. After three subcultures, cell viability evaluation was performed by Neubauer chamber quantification. Neural differentiation was induced in neurobasal culture medium (Gibco ™), with cells adhered to the plastic or suspended in agarose-treated plates. After seven and 14 days in inducer culture, morphology and immunophenotypic profile were observed using flow cytometry and fluorescent immunocytochemistry. At 14 days the cells had a high degree of expression for anti-nestin and anti-glial fibrillary acidic (anti-GFAP) markers. Previously, an average of 18x10⁶ undifferentiated viable cells from the pulp tissue were obtained on the 25th day. It is suggested that the undifferentiated canine pulp stem cells present satisfactory differentiation indices in neural progenitor cells, adhered or suspended in culture. The dental pulp of deciduous canine teeth provides viable undifferentiated cells in adequate quantity.

O objetivo deste trabalho foi estudar a produção e o valor nutritivo de nove espécies de leguminosas forrageiras herbáceas tropicais, cultivadas sob dois níveis de irradiação luminosa e seis frequências de corte na estação chuvosa, em Anhembi (SP), Brasil. Parcelas puras foram estabelecidas ao sol e sob plantação de Eucalyptus grandis, com 8 anos de idade, com 1.000 árvores/ha, que interceptou mais de 80% da radiação luminosa incidente. Avaliou-se a taxa de acúmulo de matéria seca, a proteína bruta e a digestibilidade in vitro da matéria seca em amostras de plantas colhidas em intervalos de 30, 60, 90 e 180 dias, com cortes adicionais aos 120 e 150 dias. Sob pleno sol, as leguminosas testadas toleraram cortes mais frequentes (intervalos de 30 - 90 dias), exceto o Lablab. Na sombra, o grau de tolerância variou conforme a espécie; em termos de produção de MS e de persistência, a Centrosema e a Puerária se destacaram dentre as demais leguminosas comparadas, quando os cortes foram realizados em intervalos de 60 dias, enquanto o Arachis e o Estilosantes foram os mais produtivos quando os cortes ocorreram em intervalos de 120 dias, mantendo o valor nutricional. O Alysicarpus e a Aeschynomene não persistiram sob o nível de sombreamento que prevaleceu nesse experimento.

The aim of this study was to evaluate the production and nutritive value of nine tropical legumes under two light intensities and six cutting regimens in the wet season, in Anhembi (SP) - Brazil. Pure stands were established in full light and under an eight-year-old eucalyptus plantation with 1.000 plants per hectare. The trees intercepted over80% of the incident radiation. Dry matter herbage accumulation, dry matter in vitro digestibility and crude protein under the cutting intervals 30, 60, 90 and 180 days, with additional cuttings at 120 and 150 days were evaluated. With the exception of Lablab, all other tested legume species tolerated higher cutting frequency under when grown under full sun light then under shade. The degree of shade-tolerance varied according to the species; Centrosema and Pueraria produced well at cutting intervals of 60 days, showing higher dry matter production and persistence while Arachis and Stylosanthes showed better results under longer cutting intervals, keeping the nutritional value, and can be indicated for differential grazing. Alysicarpus and Aeschynomene did not persist under the prevailing conditions.

Exaggerated blood pressure response (EBPR) during the exercise treadmill test (ETT) has been considered to be a risk factor for hypertension. The relationship of polymorphisms of the renin-angiotensin system gene with hypertension has not been established. Our objective was to evaluate whether EBPR during exercise is a clinical marker for hypertension. The study concerned a historical cohort of normotensive individuals. The exposed individuals were those who presented EBPR. At the end of the observation period (41.7 months = 3.5 years), the development of hypertension was analyzed within the two groups. Genetic polymorphisms and blood pressure behavior were assessed as independent variables, together with the classical risk factors for hypertension. The I/D gene polymorphism of the angiotensin-converting enzyme and M235T of angiotensinogen were ruled out as risk factors for hypertension. EBPR during ETT is not an independent influence on the chances of developing hypertension. No differences were observed between the hypertensive and normotensive individuals regarding gender (P = 0.655), skin color (P = 0.636), family history of hypertension (P = 0.225), diabetes mellitus (P = 0.285), or hypertriglyceridemia (P = 0.734). The risk of developing hypertension increased with increasing body mass index (BMI) and advancing age. The risk factors, which independently influenced the development of hypertension, were age and BMI. EBPR did not constitute an independent risk factor for hypertension and is probably a preclinical phase in the spectrum of normotension and hypertension.

Determinou-se a área transversal (AT) dos tendões e ligamentos da região metacárpica palmar de 15 cavalos da raça Mangalarga Marchador. As áreas transversais do tendão do músculo flexor digital superficial (TFDS), do tendão do músculo flexor digital profundo (TFDP), do ligamento acessório inferior (LA) e do ligamento suspensório (LS) foram determinadas mediante ultra-sonografia, em sete zonas distintas (1A, 1B, 2A, 2B, 3A, 3B e 3C). Os valores médios da AT para TFDS foram: zona 1A, 1,07±0,13; zona 1B, 0,97±0,12; zona 2A, 0,77±0,08; zona 2B, 0,68±0,12; zona 3A, 0,74±0,10; zona 3B, 0,86±0,10; e zona 3C, 1,33±0,24. Para TFDP foram: zona 1A, 1,09±0,12; zona 1B, 0,99±0,14; zona 2A, 0,72±0,11; zona 2B, 0,63±0,10; zona 3A, 0,71±0,14; zona 3B, 0,87±0,14; e zona 3C, 1,39±0,20. Para LA foram: zona 1A, 1,17±0,14; zona 1B, 0,85±0,11; zona 2A, 0,73±0,12; zona 2B, 0,60±0,16; e zona 3A, 0,42±0,15. Para LS foram: zona 1A, 0,82±0,14; zona 1B, 1,23±0,15; zona 2A, 1,21±0,15; zona 2B, 1,18±0,14; e zona 3A, 1,20±0,31. A AT de cada estrutura avaliada foi similar entre membros torácicos, considerando a mesma zona estudada. Não houve diferença significativa entre a AT do TFDS e a do TFDP.

The transversal area (TA) of tendons and ligaments located in the palmar metacarpal region was studied in 15 Mangalarga Marchador horses without locomotor pathology. The superficial digital flexor tendon (SDFT), deep digital flexor tendon (DDFP), accessory ligament of the deep digital flexor tendon (AL-DDFP), and suspensory ligament (TIOM) were examined by ultrasonography, at seven different zones (1A, 1B, 2A, 2B, 3A, 3B e 3C), to determine the TA. The mean TA values for SDFT were: zone 1A, 1.07±0.13; zone 1B, 0.97±0.12; zone 2A, 0.77±0.08; zone 2B, 0.68±0.12; zone 3A, 0.74±0.10; zone 3B, 0.86±0.10; and zone 3C, 1.33±0.24. For DDFT were: zone 1A, 1.09±0.12; zone 1B, 0.99±0.14; zone 2A, 0.72±0.11; zone 2B, 0.63±0.10; zone 3A, 0.71±0.14; zone 3B, 0.87±0.14; and zone 3C, 1.39±0.20. For AL-DDFT were: zone 1A, 1.17±0.14; zone 1B, 0.85±0.11; zone 2A, 0.73±0.12; zone 2B, 0.60±0.16; and zone 3A, 0.42±0.15. For TIOM were: zone 1A, 0.82±0.14; zone 1B, 1.23±0.15; zone 2A, 1.21±0.15; zone 2B, 1.18±0.14; and zone 3A, 1.20±0.31. The TA of each structure examined was similar between the thoracic members in each of the studied zones. No significant difference was found among the TA of the SDFT and that of the DDFP.