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Background Maspin (SERPINB5) is a potential tumor suppressor gene with pleiotropic biological activities, including regulation of cell proliferation, death, adhesion, migration and gene expression. Several studies indicate that nuclear localization is essential for maspin tumor suppression activity. We have previously shown that the EGFR activation leads to maspin nuclear localization in MCF-10A cells. The present study investigated which EGFR downstream signaling molecules are involved in maspin nuclear localization and explored a possible role of cell–cell contact in this process. Methods MCF-10A cells were treated with pharmacological inhibitors against EGFR downstream pathways followed by EGF treatment. Maspin subcellular localization was determined by immunofluorescence. Proteomic and interactome analyses were conducted to identify maspin-binding proteins in EGF-treated cells only. To investigate the role of cell–cell contact these cells were either treated with chelating agents or plated on different cell densities. Maspin and E-cadherin subcellular localization was determined by immunofluorescence. Results We found that PI3K-Akt and JAK2-STAT3, but not MAP kinase pathway, regulate EGF-induced maspin nuclear accumulation in MCF-10A cells. We observed that maspin is predominantly nuclear in sparse cell culture, but it is redistributed to the cytoplasm in confluent cells even in the presence of EGF. Proteomic and interactome results suggest a role of maspin on post-transcriptional and translation regulation, protein folding and cell–cell adhesion. Conclusions Maspin nuclear accumulation is determined by an interplay between EGFR (via PI3K-Akt and JAK2-STAT3 pathways) and cell–cell contact.
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RESUMO A família Annonaceae possui representantes de grande interesse medicinal e o gênero Xylopia é um dos que merecem destaque. Composta por aproximadamente 160 espécies distribuídas na América do Sul, América central, África e Ásia, as espécies desse gênero podem ser arbustivas ou arbóreas. No Brasil são encontradas nas regiões Norte, Nordeste, Centro-Oeste e Centro Sul. Este gênero produz uma variedade de metabólitos incluindo alcalóides, amidas, lignóides, acetogeninas e terpenóides e têm sido investigados como fonte potencial de acetogeninas, compostos esses que apresentam uma ampla variedade de propriedades biológicas com destaque para: citotóxica, antitumoral, antiparasitária, antimicrobial, inseticida e antimalarial. Neste estudo, efetuou-se uma revisão das principais espécies de Xylopiaencontradas no Brasil, já estudadas e descritas na literatura, abordando os aspectos químico-farmacológicos, destacando os constituintes químicos isolados bem como a ação farmacológica evidenciada.
ABSTRACT The family Annonaceae has representatives of great medical interest, and the Xylopia species deserves attention. The Xylopia genus is composed by approximately 160 species, with geographic distribution in tropical and subtropical regions of America, Africa and Asia. This genus can present shrubs or trees. In Brazil, they can be found at the North, North-west, Central-West and Central-South Regions. The phytochemical investigations resulted mainly in the isolation of alkaloids, diterpenos, quinolines and acetogenins, with the latter presenting very interesting biological properties such as the cytotoxic, antiprotozoal and the insecticide activities.This study aimed to review the botanical, chemical and pharmacological aspects of the Xylopia genus found in Brazil, highlighting the chemical components, as the well-known pharmacological effect .
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Química , Xylopia/metabolismo , Medicina Tradicional/instrumentaçãoRESUMO
Estradiol participates in the control of energy homeostasis, as demonstrated by an increase in food intake and in body weight gain after ovariectomy in rats. In the present study, female Wistar rats (200-230 g, N = 5-15 per group), with free access to chow, were individually housed in metabolic cages. We investigated food intake, body weight, plasma leptin levels, measured by specific radioimmunoassay, and the hypothalamic mRNA expression of orexigenic and anorexigenic neuropeptides, determined by real-time PCR, in ovariectomized rats with (OVX+E) and without (OVX) estradiol cypionate treatment (10 µg/kg body weight, sc, for 8 days). Hormonal and mRNA expression were determined at pre-feeding and 4 h after food intake. OVX+E rats showed lower food intake, less body weight gain and lower plasma leptin levels. In the OVX+E group, we also observed a reduction of neuropeptide Y (NPY), agouti-related protein (AgRP) and cocaine- and amphetamine-regulated transcript (CART) mRNA expression in the arcuate nucleus and a decrease in orexin A in the lateral hypothalamic area (LHA). There was an increase in leptin receptor (LepRb), melanocortin-4 receptor (MC4-R), CART, and mainly corticotropin-releasing hormone (CRH) mRNA in the paraventricular nucleus and LepRb and CART mRNA in the LHA. These data show that hypophagia induced by estradiol treatment is associated with reduced hypothalamic expression of orexigenic peptides such as NPY, AgRP and orexin A, and increased expression of the anorexigenic mediators MC4-R, LepRb and CRH. In conclusion, estradiol decreases food intake, and this effect seems to be mediated by peripheral factors such as leptin and the differential mRNA expression of neuropeptides in the hypothalamus.
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Animais , Feminino , Ratos , Ingestão de Alimentos/efeitos dos fármacos , Estradiol/farmacologia , Estrogênios/farmacologia , Hipotálamo/efeitos dos fármacos , Neuropeptídeos/metabolismo , RNA Mensageiro/metabolismo , Regulação da Expressão Gênica , Hipotálamo/química , Neuropeptídeos/genética , Ovariectomia , Reação em Cadeia da Polimerase , Ratos WistarRESUMO
Human infection with fish parasites can result from the ingestion of incompletely cooked or raw fish, giving origin to parasitic diseases such as anisakiasis, caused by parasites of the Anisakidae family. The present study assessed the in vitro larvicidal effect of two monoterpene compounds, geraniol and citronellal, against Contracaecum sp (Nematoda: Anisakidae). Four hundred live larvae of Contracaecum sp obtained from "traíra" fish (Hoplias malabaricus, Bloch, 1974) were analyzed on 40 Petri dishes (10 larvae each) with the compounds to be tested. The final concentrations tested for each compound were 250, 125, 62.5, and 31.2 µg/mL and the evaluation was carried out at five different times (2, 4, 8, 24, and 48 h). The larvicidal action of geraniol and citronellal was statistically superior (P < 0.005) to the control (1 percent ethanol) at concentrations of 250 and 31.2 µg/mL (geraniol) and 250, 125, and 62.5 μg/mL (citronellal). However, no larvicidal activity was observed at concentrations of 125 and 62.5 µg/mL for geraniol and 31.2 µg/mL for citronellal. When the larvicidal action of geraniol was compared to that of citronellal, the former was found to be statistically superior (P < 0.05) to the latter at concentrations of 250 and 31.2 μg/mL. On the other hand, citronellal was statistically superior (P < 0.005) to geraniol at concentrations of 125 and 62.5 μg/mL. The larval mortality rate in terms of time (hours) was higher for geraniol with the passing of time at the 250 μg/mL concentration. At this concentration (in 48 h) the best larvicidal effect was observed with 90 percent lethality. The larvae were considered to be dead using no motility and loss of structural integrity as parameters. The data indicate that natural terpene compounds should be more explored for antihelminthic activity and can be useful for other studies about anisakiasis treatment.
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Animais , Aldeídos/farmacologia , Antinematódeos/farmacologia , Monoterpenos/farmacologia , Nematoides/efeitos dos fármacos , Terpenos/farmacologia , Avaliação Pré-Clínica de Medicamentos , Larva/efeitos dos fármacos , Nematoides/crescimento & desenvolvimento , Testes de Sensibilidade ParasitáriaRESUMO
The Epstein-Barr virus (EBV) is the etiological agent of oral hairy leukoplakia (OHL), an oral lesion with important diagnostic and prognostic value in acquired immunodeficiency disease syndrome. The two EBV genotypes, EBV-1 and EBV-2, can be distinguished by divergent gene sequences encoding the EBNA-2, 3A, 3B, and 3C proteins. The purpose of this study was to identify the EBV genotype prevalent in 53 samples of scrapings from the lateral border of the tongue of HIV-1 seropositive patients, with and without OHL, and to correlate the genotypes with presence of clinical or subclinical OHL with the clinic data collected. EBV-1 and EBV-2 were identified through PCR and Nested-PCR based on sequence differences of the EBNA-2 gene. EBV-1 was identified in the 31 samples (15 without OHL, 7 with clinical OHL and 9 with subclinical OHL), EBV-2 in 12 samples (10 without OHL, 1 with clinical and 1 subclinical OHL), and a mixed infection in 10 samples (2 without OHL, 3 with clinical and 5 with subclinical OHL). The presence of EBV-1 was higher in women, but a significant statistical result relating one the EBV genotypes to the development of OHL was not found. We conclude that the oral epithelium in HIV-1 seropositive patients can be infected by EBV-1, EBV-2 or by a mixed viral population.
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Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infecções Oportunistas Relacionadas com a AIDS/virologia , HIV-1 , /genética , Leucoplasia Pilosa/virologia , Língua/virologia , DNA Viral/genética , Eletroforese em Gel de Ágar , Genótipo , /classificação , Reação em Cadeia da PolimeraseRESUMO
This paper relates the clinical and epidemiological aspects of canine parvovirus infection (CPV) in the State of Rio de Janeiro from April 1995 to March 2004. A total of 341 fecal samples were collected from up to 6-months-old puppies with gastroenteritis. The diagnosis of CPV infection was confirmed by hemagglutination/ hemagglutination inhibition tests, enzyme immunoassay, virus isolation in cell culture or polymerase chain reaction. One hundred and fifty-seven samples (46 percent) were positive for CPV. No correlation among sex, breed or age and the occurrence of CPV infection was observed. The classical signs of parvoviral enteritis (anorexia, lethargy, vomiting and hemorrhagic fluid diarrhea) were observed in 70 percent of CPV-positive and in 60 percent of CPV-negative puppies. Although CPV could be detected throughout the studied period, its occurrence was significantly higher from June to September and November to December. These results show that CPV is still circulating in the State of Rio de Janeiro.
Este trabalho relata os aspectos clínicos e epidemiológicos da infecção pelo CPV no Estado do Rio de Janeiro, no período de abril de 1995 a março de 2004. Coletaram-se 341 amostras fecais de cães com até seis meses de idade que apresentavam gastrenterite. O diagnóstico da infecção pelo CPV foi confirmado através dos testes de hemaglutinação/inibição da hemaglutinação, ensaio imunoenzimático, isolamento viral em cultura de células ou reação em cadeia pela polimerase. Cento e cinqüenta e sete amostras (46 por cento) foram consideradas positivas para CPV. Não foi observada correlação entre sexo, raça ou idade e a ocorrência da infecção por CPV. Os sinais clínicos clássicos de parvovirose (vômito, anorexia, apatia e diarréia líquida hemorrágica) foram observados em 70 por cento dos animais positivos e 60 por cento dos animais negativos para CPV. O CPV foi detectado ao longo do período estudado, entretanto observou-se um aumento do número de casos positivos nos períodos de junho a setembro e novembro a dezembro. Estes resultados mostram que o CPV ainda circula no Estado do Rio de Janeiro.
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Animais , Coliformes/análise , Cães , Enterite/diagnóstico , Enterite/epidemiologia , Enterite/prevenção & controle , Parvovirus Canino/isolamento & purificaçãoRESUMO
Avaliou-se a difusão da infecção em um rebanho com prévio isolamento de Salmonella sp, em que leitões, individualmente identificados, foram amostrados para excreção fecal de Salmonella sp e sorologia do nascimento ao abate. Da mesma forma, amostras de ração e suabes do ambiente foram coletados durante o estudo para pesquisa de Salmonella sp A pesquisa de anticorpos foi realizada pela utilização de ELISA-LPS de Salmonella Typhimurium. Os leitões foram negativos na análise bacteriológica e na sorologia até a fase de creche, tornando-se positivos para Salmonella sp no início da terminação. Nessa amostragem, 28,6 por cento dos animais foram soropositivos e 75 por cento estavam excretando Salmonella sp nas fezes. Ao abate, a percentagem de animais soropositivos (76,9 por cento) aumentou, enquanto o isolamento de Salmonella sp ocorreu em 19,2 por cento dos suínos. Foi isolada Salmonella sp de duas das 26 amostras de ração. A contaminação do ambiente da terminação ocorreu apenas após o alojamento dos animais. Concluiu-se que a terminação foi o ponto crítico de contaminação desse lote, sendo a ração uma fonte de contaminação.
Salmonella diffusion in a swine production system, previously identified as Salmonella-positive, was evaluated. A cohort of pigs was followed from farrowing to slaughtering. Samples of feces, intestinal content, mesenteric lymph nodes and blood were taken from the animals throughout the study. In addition, feed samples and environmental swabs were done for the isolation of Salmonella sp. Serum was submitted to a Salmonella Typhimurium LPS-ELISA. Piglets were negative in bacteriological and serological tests until the nursery phase, but became Salmonella positive in the early finishing. On this phase, 28.6 percent of finishers were seropositive and 75 percent were shedding Salmonella in feces. At slaughtering, the seropositivity (76.9 percent) was higher than in the early finishing, but Salmonella was isolated only from 19.2 percent of the sampled pigs. Two out of 26 feed samples were Salmonella positive. Contamination of the finishing site environment was detected only when the animals were housed. It was concluded that the termination phase was critical for the contamination of this cohort of pigs, being the feed a source of contamination.