Effects of nemorosone, isolated from the plant Clusia rosea, on the cell cycle and gene expression in MCF-7 BUS breast cancer cell lines

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mercury fractionation in dourada (Brachyplatystoma rousseauxii) of the Madeira River in Brazil using metalloproteomic strategies

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The maspin expression in canine mammary tumors: an immunohistochemical and molecular study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação da contratilidade da vesícula biliar com leite materno e leite de vaca em lactentes

As múltiplas características distintas do leite materno e do leite de vaca fazem com que o esvaziamento gástrico seja mais rápido após ingestão do primeiro. Esta rapidez do esvaziamento gástrico pode estar associada ao esvaziamento da vesícula biliar, também mais rápido. O objetivo do trabalho foi determinar se há diferença na velocidade de esvaziamento da vesícula biliar, por método ultra-sonográfico, após ingestão de leite materno e leite de vaca. Estudamos 47 crianças menores de dois anos, 22 após uma refeição com leite materno e 25 após uma refeição com leite de vaca. A vesícula biliar foi medida após um período de quatro a seis horas de jejum e uma hora após uma refeição (leite materno ou leite de vaca), e a contração da vesícula biliar foi avaliada, em porcentagem, pela diferença de volume da vesícula biliar entre as duas medidas e expressa em porcentagem do volume inicial. As medianas das porcentagens do volume inicial para as crianças alimentadas com leite materno e leite de vaca foram, respectivamente, 92,25% e 53,8%, diferença estatisticamente significativa. Portanto, a contração da vesícula biliar após uma refeição com leite de vaca é muito mais lenta do que aquela após uma refeição com leite materno.

Diagnosis of clinical bovine mastitis by fine needle aspiration followed by staining and scanning electron microscopy in a Prototheca zopfii outbreak

Biopsy by fine needle aspiration together with microbiological examination and scanning electron microscopy were evaluated in diagnosis of clinical bovine mastitis in a Prototheca zopfii outbreak. Fine needle aspiration was performed in 21 mammary quarters from ten Holstein cows presenting clinical mastitis caused by P. zopfii. The algae were previously identified in the microbiological examination of milk collected from these cows. Material aspirated from these 21 mammary glands was submitted to cytological staining (Gram, Giemsa and/or Shor staining). Fine needle aspiration enabled cytological identification of the algae in these 21 mammary glands, from which P. zopfii was isolated in the milk. Simultaneously, five mammary fragments collected by fine needle aspiration from these 21 mammary glands presenting clinical mastitis were also submitted to microbiological examination. P. zopfii was also isolated from these five fragments. Scanning electron microscopy technique also identified three of these five P. zopfii strains isolated from mammary fragments collected by cytological aspiration. These results suggest that fine needle aspiration may be an alternative method for the diagnosis of clinical mastitis.

Somaticell® as a screening method for somatic cell count from bovine milk

Os objetivos do presente estudo foram avaliar a correlação entre a contagem eletrônica de células somáticas (eCCS) com o Somaticell® sob diferentes níveis de contagem de células somáticas (CCS) do leite e patógenos causadores de mastites, além de calcular a sensibilidade, especificidade e valores preditivos do Somaticell® utilizando diferentes limites de CCS estabelecidos pelos diferentes países. Trezentos e quarenta amostras de leite foram coletadas assepticamente após realização do California Mastitis Test (CMT). O Somaticell® e a eCCS foram realizados em todas as amostras de leite. A correlação entre o Somaticell® e a contagem eletrônica foi determinada de acordo com o CMT, patógeno isolado e escore de eCCS. de acordo com os escores de CCS estabelecidos, 26,5% das amostras de leite apresentaram escore 1 (69-166 x10³células mL-1), 26,8% escore 2 (167-418x10³células mL-1), 27,4% escore 3 (419-760x10³células mL-1) e 19,4% escore 4 (761 to 1970x10³células mL-1). A eCCS e o Somaticell® apresentaram correlação positiva em quase todos os escores estudados (exceto escore 2 e 3). O valor de r obtido entre CCS e o Somaticell® foi de 0,32. Observou-se que, quando o limite de CCS estabelecido aumentou, a sensibilidade decresceu e os valores de especificidade aumentaram. Os valores preditivos apresentaram-se constantes em todos os limites. Quando o limite de CCS era baixo (<760,000 células mL-1), Somaticell® forneceu resultados consistentemente mais elevados que os valores de CCS. Já para amostras com CCS elevada, Somaticell® resultou em menores contagens que a eCCS. A correlação entre os dois métodos permaneceu relativamente constante em todas as condições e os valores de sensibilidade e especificidade do teste são altamente dependentes do limite estabelecido. Os resultados deste trabalho sugerem que o Somaticell® não é útil para avaliar a CCS do leite, pois seus resultados são significativamente diferentes da eCCS, no entanto, pode ser utilizado como método de triagem, tal como o CMT, para a detecção do aumento da CCS do leite.

Traceability of bovine meat and bone meal in poultry by stable isotope analysis

Bovine meat and bone meal (MBM) was widely used in animal diets until outbreaks of Bovine Spongiform Encefalopathy (BSE) occurred in some countries. It has not been confirmed yet whether or not BSE may be transmitted to man through chicken meat originated from poultry that had been fed diets containing MBM. Therefore, consumers nowadays express preference for meat originated from birds fed exclusively vegetable diets. This study analyzed samples of major breast muscle (Pectoralis major) using mass spectrometry of stable isotopes (carbon and nitrogen) as a means to assess the presence of MBM in broiler diets, a technique that might be used in the certification of poultry quality. A total of 150 day-old chicks were reared in five randomized treatments with increasing MBM dietary inclusion levels (0, 1, 2, 4 and 8%). on day 42, breast muscle samples were collected from three birds per treatment and used in the determination of 13C/12C and 15N/14N isotope ratios. The breast muscle isotope values were expressed as delta in parts per thousand (delta ). The following carbon isotope values (13C) were found: 18.74 ±0.11, 18.51 ±0.19, 18.24 ±0.10, -17.79 ±0.12, and -17.15 ±0.15 for 0, 1, 2, 4 and 8% MBM dietary levels, respectively. Nitrogen isotope values (15N) were 1.65 ±0.14, 1.65 ±0.28, 1.72 ±0.08, 1.95 ±0.16, and 2.52 ± 0.09 for 0, 1, 2, 4 and 8% MBM dietary levels, respectively. This study showed important differences in delta13C and delta15N values in breast meat, evidencing a simultaneous enrichment of this isotopic pair, which allowed tracing MBM in bird diets. Analysis of carbon and nitrogen stable isotopes may be used to ensure feeding with exclusively vegetable diets, and might also be used as a reliable evaluation tool in broiler meat certification. The diet with 1% inclusion level of MBM and the exclusively vegetable diet showed similar results.

Clinical Approach of Ankyloglossia in Babies: Report of Two Cases

Ankyloglossia is a developmental anomaly of the tongue characterized by a short lingual frenum, resulting in restricted movement of the tongue. Its etiology is undefined and there is no gender preference. Few studies are available in the literature and the diagnosis and management of ankyloglossia in infants remains controversial. We report two cases of infants submitted to lingual frenectomy, emphasizing the management of ankyloglossia and its implications in breast-feeding.

Influence of estradiol and triiodothyronine on breast cancer cell lines proliferation and expression of estrogen and thyroid hormone receptors

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of Gestational and Lactational Fenvalerate Exposure on Immune and Reproductive Systems of Male Rats

The aim of this study was to determine the consequent reproductive developmental and immunotoxic effects due to exposure to fenvalerate during pregnancy and lactation in male offspring of maternal-treated rats. Pregnant rats were treated daily by oral gavage with 40 or 80 mg/kg of fenvalerate or corn oil (vehicle, control), from d 12 of pregnancy to d 21 of lactation. Immune and reproductive developmental effects were assessed in male offspring at postnatal days (PND) 40 (peripuberty), 60 (postpuberty), and 90 (sexual maturity). Treatment with the higher dose (80 mg/kg) resulted in convulsive behavior, hyperexcitability, and mortality in 45% of the dams. Fenvalerate was detected in the fetus due to placental transfer, as well as in pups due to breast-milk ingestion, persisting in male offspring until PND 40 even though pesticide treatment was terminated on PND 20. However, fenvalerate did not produce marked alterations in age of testicular descent to the scrotum and prepucial separation, parameters indicative of puberty initiation. In contrast, at puberty, there was a reduction in testicular weight and sperm production in male offspring of maternal-treated rats. At adulthood, the sperm counts and fertility did not differ between control and treated groups. Testosterone levels were not changed at any time during reproductive development. Similarly, no apparent exposure-related effects were detected in the histological structures of the lymphohematopoietic system. Data indicate that fenvalerate, in this experimental model, interfered with initial development of the male reproductive system, but that these effects on sperm production or fertility did not persist into adulthood. There was no apparent evidence that fenvalerate altered testosterone levels or produced a disruption in male endocrine functions.

Association of matrix metalloproteinase-8 gene variation with breast cancer prognosis

Animal and cell studies indicate an inhibitory effect of matrix metalloproteinase-8 (MMP8) on tumorigenesis and metastasis. We investigated whether MMP8 gene variation was associated with breast cancer metastasis and prognosis in humans. We first studied nine tagging single nucleotide polymorphisms (SNP) in the MMP8 gene in 140 clinically and pathologically well-characterized breast cancer patients. Four of the SNPs were found to be associated with lymph node metastasis, the most pronounced being a promoter SNP (rs11225395) with its minor allele (T) associating with reduced susceptibility to lymph node metastasis (P = 0.02). This SNP was further evaluated for association with cancer relapse and survival among a cohort of similar to 1,100 breast cancer patients who had been followed for cancer recurrence and mortality for a median of 7.1 years. The T allele was associated with reduced cancer relapse and greater survival, particularly among patients with earlier stage cancer. Among patients of tumor-node-metastasis stage 0 to 11, the adjusted hazard ratio of disease-free survival was 0.7 [95% confidence interval (95% CI), 0.5-0.9] for patients carrying T allele compared with those homozygous for the C allele (P = 0.02). In vitro experiments showed that the T allele had higher promoter activity than the C allele in breast cancer cells. Electrophoretic mobility shift assays showed binding of nuclear proteins to the DNA sequence at the SNP site of the T allele but not that of the C allele. The data suggest that MMP8 gene variation may influence breast cancer prognosis and support the notion that MMP8 has an inhibitory effect on cancer metastasis.

Determination of Hg in water by CVAAS using 2-aminothiazole modified silica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cara inchada of cattle, an infectious, apparently soil antibiotics-dependant periodontitis in Brazil

O objetivo desta revisão das pesquisas sobre a cara inchada dos bovinos (CI), realizadas no decorrer dos últimos 30 anos, é de elucidar melhor a sua etiologia. A CI geralmente tem sido considerada de origem nutricional, causada primariamente por deficiência ou desequilíbrio mineral. A doença caracteriza-se por uma periodontite rapidamente progressiva, que afeta os tecidos peridentários a nível dos premolares e molares no período de erupção dos dentes e que se inicia geralmente em bezerros jovens. A doença causou grandes perdas econômicas aos pecuaristas da Região Centro-Oeste do Brasil, nas décadas de 1960 e 1970, com a ocupação de novas terras para criação de gado. O freqüente abaulamento lateral dos ossos maxilares nos bezerros, que deu à doença o nome popular de cara inchada, foi demonstrado ser conseqüente à periostite crônica ossificante resultante da alveolite purulenta da CI. Das lesões peridentárias foi isolado, em grande número, Bacteroides melaninogenicus, sempre junto com Actinomyces (Corynebacterium) pyogenes. Bactérias classificadas como pertencentes ao grupo sacarolí-tico e não-sacarolítico dos pigmentados de negro Bacteroides melaninogenicus e Bacteroides spp também foram isoladas, em pequeno número, de bovinos jovens sadios de fazendas CI-negativas. Ensaios in vitro mostraram que os antibióticos estreptomicina e actinomicina, bem como os sobrenadantes de cultivos de actinomicetos do solo de fazendas CI-positivas, aplicadas nas bactérias ensaiadas em concentrações subinibidoras, aumentaram significativamente (até 10 vezes) a aderência de B.melaninogenicus a células epiteliais da gengiva bovina. Esses antibióticos são produzidos no solo em conseqüência de um aumento do número de actinomicetos, incluindo os do gênero Streptomyces, quando há modificação de sua microbiota em áreas previamente ocupadas por mata virgem ou vegetação natural de Cerrado, que foram cultivadas pela primeira vez na formação de pastagem para o gado. em face da epidemiologia da CI, há fortes evidências de que a ingestão desses antibióticos pelos bovinos, junto com a forrageira, seja importante fator desencadeante para o desenvolvimento da periodontite. Através do aumento da aderência de B. melaninogenicus ao epitélio da gengiva marginal, em face da ingestão dos antibióticos pelos animais, as bactérias conseguem colonizar, formar a placa bacteriana e tornar-se patogênicas. Há evidência de que o fator desencadeante (aparentemente, os antibióticos) esteja também presente no leite de vacas-mães de bezerros afetados pela CI. Foi demonstrado que as bactérias envolvidas na periodontite produzem enzimas e endotoxinas capazes de ação destrutiva sobre os tecidos peridentários. A epidemiologia da CI, com a diminuição de sua incidência e o seu desaparecimento no decorrer dos anos, pode ser explicada pelo fato de que o prévio equílibrio da microbiota no solo virgem foi alcançado novamente e a produção dos antibióticos se reduziu. Desta maneira, a CI deve ser considerada como uma periodontite infecciosa multifatorial, causada sobretudo por bactérias anaeróbias pertencentes ao grupo Bacteroides melaninogenicus e, ao que tudo indica, desencadeada pela ingestão contínua, com a forrageira, de concentrações subinibidoras de antibióticos de solos recentememente cultivados. Esta hipótese é reforçada pela observação recente de novos surtos de CI, em áreas anteriormente positivas para a doença, em conseqüência da reforma de pastagens e capineiras após muitos anos. A natureza infecciosa da CI-periodontite foi confirmada através de experimento, em que virginiamicina mostrou-se eficaz no tratamento oral de bovinos afetados pela doença. Os antibióticos espiramicina e virginiamicina, usados como aditivos em suplementos minerais no campo, mostraram-se eficientes na prevenção da CI.

Fast ultrasound-assisted treatment of urine samples for chronopotentiometric stripping determination of mercury at gold film electrodes

This work describes an efficient, fast, and reliable analytical methodology for mercury determination in urine samples using stripping chronopotentiometry at gold film electrodes. The samples were sonicated in the presence of concentrated HCl and H2O2 for 15 min in order to disrupt the organic ligands and release the mercury. Thirty samples can be treated over the optimized region of the ultrasonic bath. This sample preparation was enough to allow the accurate stripping chronopotentiometric determination of mercury in the treated samples. No background currents and no passivation of the gold film electrode due to the sample matrix were verified. The samples were also analyzed by cold vapour atomic absorption spectrometry (CV-AAS) and good agreement between the results was verified. The analysis of NIST SRM 2670 (Toxic Metals in Freeze-Dried Urine) also validated the proposed electroanalytical method. Finally, this method was applied for mercury evaluation in urine of workers exposed to hospital waste incinerators. (c) 2006 Elsevier B.V. All rights reserved.

Synchronization rate, size of the ovulatory follicle, and pregnancy rate after synchronization of ovulation beginning on different days of the estrous cycle in lactating dairy cows

Recently a protocol was developed that precisely synchronizes the time of ovulation in lactating dairy cows (Ovsynch; GnRH-7d-PGF(2 alpha)-2d-GnRH). We evaluated whether initiation of Ovsynch on different days of the estrous cycle altered the effectiveness of this protocol. The percentage of cows (n=156) ovulating to the first GnRH was 64% and varied (P<0.01) by stage of estrous cycle. Treatment with PGF(2 alpha) was effective, with 93% of cows having low progesterone at second GnRH. The overall percentage of cows that ovulated after second GnRH (synchronization rate) was 87% and varied by response to first GnRH (92% if ovulation to first GnRH vs 79% if no ovulation; P<0.05). There were 6% of cows that ovulated before the second injection of GnRH and 7% with no detectable ovulation by 48 h after second GnRH. Maximal diameter of the ovulatory follicle varied by stage of estrous cycle, with cows in which Ovsynch was initiated at midcycle having the smallest follicles. In addition, milk production and serum progesterone concentration on the day of PGF(2 alpha) affected (P<0.05) size of the ovulatory follicle. Using these results we analyzed pregnancy rate at Days 28 and 98 after Al for cows (n=404) in which Ovsynch was initiated on known days of the estrous cycle. Pregnancy rate was lower for cows expected to ovulate larger follicles than those expected to ovulate smaller follicles (P<0.05; 32 vs 42%). Thus, although overall synchronization rate with Ovsynch was above, 85%, there were clear differences in response according to day of protocol initiation. Cows in which Ovsynch was initiated near midcycle had smaller ovulatory follicles and greater pregnancy rates. (C) 1999 by Elsevier B.V.