Implications on the Pb bioaccumulation and metallothionein levels due to dietary and waterborne exposures: The Callinectes danae case.

This study aimed to assess the bioaccumulation of Pb and induction of metallothionein-like proteins (MT) in Callinectes danae through single and combined dietary and waterborne exposures. Male C. danae individuals were collected in the south area of the Cananéia-Iguape-Peruíbe Protected Area (APA-CIP), in São Paulo State, Brazil. After an acclimatization period, exposure assays were performed during 7 and 14 days, at two Pb concentrations (0.5 e 2.0 µg/g) in 4 treatments: 1) control; 2) contaminated water only; 3) contaminated food only; 4) contaminated water and food. The results indicate that C. danae is highly tolerant to Pb exposure at the evaluated concentrations. In gills, Pb bioaccumulation is more dependent of water efflux and time of exposure (higher Pb values). However, pathways act simultaneously in the induction of MT expression in this tissue. The decreases in Pb accumulation in the combined treatments and MT increases after 14 days in gills suggests that these proteins play a detoxification function in the presence of Pb. In hepatopancreas, depending on the predominance of a certain pathway or combined pathways, accumulation occured at different times. For muscle tissue, bioaccumulation was observed due to contaminated water exposure, but not dietary exposure, probably because Pb concentrations were low.

Morphology and histology of vom Rath's organ in brush-footed butterflies (Lepidoptera: Nymphalidae).

Vom Rath's organ, located at the distal end of the third segment of the labial palp, is one of the recognized synapomorphies of Lepidoptera (Insecta). Information about the structural and histological morphology of this organ is sparse. The structure of vom Rath's organ in four species of Nymphalidae, three frugivorous: Fountainea ryphea (Charaxinae: Anaeini), Morpho helenor achillaena (Satyrinae: Morphini) and Hamadryas epinome (Biblidinae: Ageroniini), and the nectarivorous species Aeria olena (Danainae: Ithomiini) is described by means of scanning electron microscopy and histology. The species showed significant differences in the cavity shape, setal morphology and arrangement, opening shape and location, associated with the organization of cell groups, type of axon, and degree of development. These differences do not seem to be related to feeding habit. No cell groups were found in Actinote thalia (Heliconiinae: Acraeini) and Heliconius erato phyllis (Heliconiinae: Heliconiini), and for the first time the absence of vom Rath's organ is documented in the clade Ditrysia. A terminology is proposed to improve understanding of the organ morphology, with an extensive analysis of the previous descriptions.

Effects of methylene blue-mediated photodynamic therapy on a mouse model of squamous cell carcinoma and normal skin.

BACKGROUND: Photodynamic therapy is used to treat a variety of cancers and skin diseases by inducing apoptosis, necrosis, immune system activation, and/or vascular damage. Here, we describe the effects of a single photodynamic therapy session using methylene blue on a mouse model of squamous cell carcinoma and normal skin. METHODS: The photodynamic therapy protocol comprised application of a 1% methylene blue solution, followed by irradiation with a diode laser for 15 min at 74 mW/cm2, for a total dose of 100 J/cm2. Morphological changes, cell proliferation, apoptosis, collagen quantity, immune system activity, and blood vessel number were analyzed 24 h and 15 days after photodynamic therapy. RESULTS: In the squamous cell carcinoma group, photodynamic therapy reduced tumor size and cell proliferation and raised cytokine levels. In normal skin, it decreased cell proliferation and collagen quantity and increased apoptosis and blood vessel numbers. CONCLUSIONS: The effects of photodynamic therapy were greater on normal skin than squamous cell carcinoma tissues. The reduced epithelial thickness and keratinization of the former are factors that contribute to the efficacy of this treatment. Adjustments to the treatment protocol are necessary to potentiate the effects for squamous cell carcinoma therapy.

Prebiotic effects of inulin extracted from burdock (Arctium lappa) in broilers / Efeito prebiótico da inulina extraída da bardana (Arctium lappa) em frangos de corte

The aim of this study was to evaluate the prebiotic effect of burdock (Arctium lappa) in commercial poultry. Four experiments were conducted to evaluate the performance parameters and the protection after challenge with Salmonella Enteritidis and Salmonella Kedougou, with and without Bifidobacterium probiotic. In two trials, the chickens were fed with flour burdock 1% during 42 days. In the other two, the chickens were fed with fructan extracted from burdock (inulin), by gavage, at a concentration of 100 mg/bird, during the first three days of life. The results showed that the broilers treated with burdock flour showed underperformed, with less weight gain from the second week, and the worst results in the fattening stage. The treated birds had diarrhea and impaired intestinal integrity. However, the groups treated with the flour had a lower rate of intestinal colonization by Salmonella Kedougou, after challenge. No statistically significant differences were detected in the performance parameters of broilers receiving the inulin, and the morphometric analysis showed no lesions in the intestinal villi. However, there was no protection in the challenge with Salmonella Enteritidis, regardless of association with probiotic. These results demonstrated that the manner of administration has influence on the prebiotic effect of burdock. The burdock flour was administered for 42 days, which may have influenced intestinal mucosal injury. Instead, the inulin was given only in the first three days, which may have been insufficient for protection against Salmonella. New experiments are needed to determine an able formulation for a protective effect, without negative impact on growth, weight gain and feed conversion of the supplemented animals.(AU)

Este projeto teve por objetivo avaliar o efeito prebiótico da bardana (Arctium lappa) em aves comerciais. Foram realizados quatro experimentos para avaliar os parâmetros zootécnicos e o grau de proteção após o desafio com Salmonella Kedougou e Salmonella Enteritidis, com e sem a adição de probióticos à base de Bifidobacterium. Em dois experimentos, as aves receberam a farinha de bardana 1% na ração, durante 42 dias. Nos outros dois, as aves receberam o frutano extraído da bardana (inulina), por gavagem, na concentração de 100 mg/ave, nos três primeiros dias de vida. Os resultados demonstraram que os frangos tratados com farinha de bardana apresentaram desempenho zootécnico inferior ao controle, com menor ganho de peso a partir da segunda semana e piores resultados na fase de engorda. As aves tratadas apresentaram diarreia e comprometimento da integridade intestinal. Em contrapartida, os grupos tratados com a farinha tiveram menor taxa de colonização intestinal por Salmonella Kedougou, após o desafio. Não foram detectadas diferenças estatisticamente significativas nos parâmetros zootécnicos dos frangos que receberam a inulina, e a análise morfométrica não evidenciou lesões nas vilosidades intestinais. No entanto, não houve proteção no desafio por Salmonella Enteritidis, independentemente da associação com probiótico. Esses resultados demonstraram que o modo de administração tem influência sobre o efeito prebiótico da bardana. A farinha de bardana foi administrada por 42 dias, o que pode ter causado a lesão da mucosa intestinal. Em contrapartida, a inulina foi administrada apenas nos primeiros três primeiros dias, o que pode ter sido insuficiente para proteção contra Salmonella. Novos experimentos são necessários para determinar uma formulação capaz de promover efeito protetor, sem impacto negativo no crescimento, ganho de peso e conversão alimentar dos animais suplementados.(AU)

Initial peptidomic profiling of Brazilian sea urchins: Arbacia lixula, Lytechinus variegatus and Echinometra lucunter

Background Sea urchins can be found throughout the Brazilian coast and are reported to be one of the major causes of marine accidents on the shoreline. Although not lethal, these accidents are reported to be extremely painful. In order to understand the toxinology of the Brazilian urchins, a peptidomic approach was performed aiming to characterize the naturally occurring peptides in both the coelomic fluid and the spine. Methods Animals were collected without gender distinction and samples of the coelomic fluid and spines extracted were analyzed by RP-HPLC and mass spectrometry for peptide de novo sequencing. Results Several peptides were identified either in the coelomic fluid or the spine extract (except for E. lucunter). The peptide sequences were aligned with public deposited sequences and possible functions were inferred. Moreover, some peptides can be cryptides, since their sequences were identified within functional proteins, for example thymosin from Strongylocentrotus purpuratus. Conclusions Although preliminary, the peptidomic approach presented here reports, for the first time, the abundance of novel biological molecules derived from these animals. The discovery of such molecules may be of potential biotechnological application, as described for other organisms; nevertheless, further studies are required.(AU)

Initial peptidomic profiling of Brazilian sea urchins: Arbacia lixula, Lytechinus variegatus and Echinometra lucunter

Sea urchins can be found throughout the Brazilian coast and are reported to be one of the major causes of marine accidents on the shoreline. Although not lethal, these accidents are reported to be extremely painful. In order to understand the toxinology of the Brazilian urchins, a peptidomic approach was performed aiming to characterize the naturally occurring peptides in both the coelomic fluid and the spine. Methods Animals were collected without gender distinction and samples of the coelomic fluid and spines extracted were analyzed by RP-HPLC and mass spectrometry for peptide de novo sequencing. Results Several peptides were identified either in the coelomic fluid or the spine extract (except for E. lucunter). The peptide sequences were aligned with public deposited sequences and possible functions were inferred. Moreover, some peptides can be cryptides, since their sequences were identified within functional proteins, for example thymosin from Strongylocentrotus purpuratus. Conclusions Although preliminary, the peptidomic approach presented here reports, for the first time, the abundance of novel biological molecules derived from these animals. The discovery of such molecules may be of potential biotechnological application, as described for other organisms; nevertheless, further studies are required.

Hepatotrophic factors reduce hepatic fibrosis in rats.

CONTEXT: Hepatic fibrosis occurs in response to several aggressive agents and is a predisposing factor in cirrhosis. Hepatotrophic factors were shown to stimulate liver growth and to restore the histological architecture of the liver. They also cause an improvement in liver function and accelerate the reversion of fibrosis before it progresses to cirrhosis. OBJECTIVE: To test the effects of hepatic fibrosis solution composed by amino acids, vitamins, glucose, insulin, glucagon and triiodothyronine on hepatic fibrosis in rats. METHODS: Fibrosis was induced in rats by gastric administration of dimethylnitrosamine (10 mg/kg) for 5 weeks. After liver biopsy, the rats received either hepatotrophic factors solution (40 mg/kg/day) or saline solution for 10 days by intraperitoneal injection. Blood samples and liver fragments were collected for hepatic function analysis, standard histopathology evaluation, and morphometric collagen quantification. RESULTS: Rats in the hepatotrophic factors group showed a decrease of the histopathological components of fibrosis and an increase of their hepatic mass (12.2%). There was no development of neoplasic lesions in both groups. Compared with the saline group, the hepatotrophic factors group also had a decrease of blood levels of hepatic-lesion markers (AST, ALT) and a decrease of collagen content in the portal spaces (31.6%) and perisinusoidal spaces (42.3%), as well as around the hepatic terminal vein (57.7%). Thus, hepatotrophic factors administration in the portal blood promoted a regenerative hepatic response, with an overall reduction of the volumetric density of collagen, improved hepatic function, and a general improvement in the histopathological aspects of fibrosis. CONCLUSION: Taken together, these results suggest the potential therapeutic use of this hepatotrophic factors solution to treat chronic liver diseases.

Adaptation of the gill epithelium of an euryhaline fish, the guppy (Poecilia vivipara), to freshwater / Adaptação do epitélio branquial de peixes eurialinos, guaru (Poecilia vivipara), para água doce

The south-american euryhaline fish Poecilia vivipara (BLOCH; SNEIDER, 1801), the guppy, is found both in estuary and river waters, which suggests high adaptability to environments of different salinity. In this work we studied the adaptation of the interlamellar, bars and rakers epithelia of the gills of estuary fish to freshwater conditions. The results reveal that the gill epithelia of Poecilia vivipara can adjust itself to freshwater by decreasing the VP of mucous cells of the interlamellar epithelium and increase the volumetric proportion (VP) of chloride cells. However, there was no evidence of similar morphological alteration in the rakers region. The epithelia of the rakers appears to be part of a different compartment that is less sensitive to variation of salinity.

O peixe eurihalino sul-americano Poecilia vivipara (BLOCH; SNEIDER, 1801), o guppy, é encontrado tanto em estuários quanto em águas de rios, o que sugere uma alta adaptabilidade aos diferentes ambientes de salinidade. Neste trabalho, estudamos a adaptação do epitélio interlamelar, do arco e do rastelo das brânquias dos peixes de estuário de água doce. Os resultados revelam que o epitélio branquial de Poecilia vivipara pode ajustar-se à água doce, diminuindo a proporção volumétrica (PV) de células mucosas do epitélio interlamelar e aumentando a PV de células clorídricas. No entanto, não houve nenhuma evidência de alteração morfológica semelhante na região do rastelo branquial. O epitélio do rastelo branquial parece ser parte de um compartimento diferente que é menos sensível a variações de salinidade.

Anatomia e histologia gastrintestinal da garoupa-verdadeira Epinephelus marginatus (Lowe, 1834) (Teleostei, Serranidae) / Dusky grouper Epinephelus marginatus (Lowe, 1834) (Teleostei, Serranidae) gastrintestinal anatomy and histology

A garoupa-verdadeira (Epinephelus marginatus) é uma espécie de serranídeo com grande importância ecológica e boas perspectivas para o cultivo. A investigação da morfologia do trato digestório, associada à atividade alimentar, é fundamental para a compreensão da biologia dessa espécie e premissa básica para pesquisas sobre requerimentos nutricionais, desenvolvimento de rações e práticas adequadas de manejo alimentar. O presente trabalho descreve a anatomia e, por meio da microscopia de luz, as variações histológicas ao longo do tubo digestório de juvenis de Epinephelus marginatus, correlacionando a histologia de cada seguimento com as respectivas funções e o hábito alimentar. Verificou-se que essa espécie possui elevado número de tipos celulares ao longo do tubo digestório que estão intimamente relacionados com a resposta imunológica inata e adaptativa, permitindo o hábito alimentar detritívoro.

The dusky grouper (Epinephelus marginatus) is a serranida species with great ecological importance and good perspectives for rearing. The investigation of the digestive tract morphology associated to the feeding activity is essential for the understanding of the biology of this species and a basic premise for research on feed requirements, feed development and appropriate feeding methods. The present study describes the anatomy and histological variations along the digestive tract of Epinephelus marginatus juveniles associating the histology with function and feeding habit. It was verified that this species possesses a high number of different cellular types along the digestive tube intimately related with the innate and adaptative immunological responses.

Hepatotrophic factors reduce hepatic fibrosis in rats / Fatores hepatotróficos reduzem a fibrose hepática em ratos

CONTEXT: Hepatic fibrosis occurs in response to several aggressive agents and is a predisposing factor in cirrhosis. Hepatotrophic factors were shown to stimulate liver growth and to restore the histological architecture of the liver. They also cause an improvement in liver function and accelerate the reversion of fibrosis before it progresses to cirrhosis. OBJECTIVE: To test the effects of hepatic fibrosis solution composed by amino acids, vitamins, glucose, insulin, glucagon and triiodothyronine on hepatic fibrosis in rats. METHODS: Fibrosis was induced in rats by gastric administration of dimethylnitrosamine (10 mg/kg) for 5 weeks. After liver biopsy, the rats received either hepatotrophic factors solution (40 mg/kg/day) or saline solution for 10 days by intraperitoneal injection. Blood samples and liver fragments were collected for hepatic function analysis, standard histopathology evaluation, and morphometric collagen quantification. RESULTS: Rats in the hepatotrophic factors group showed a decrease of the histopathological components of fibrosis and an increase of their hepatic mass (12.2 percent). There was no development of neoplasic lesions in both groups. Compared with the saline group, the hepatotrophic factors group also had a decrease of blood levels of hepatic-lesion markers (AST, ALT) and a decrease of collagen content in the portal spaces (31.6 percent) and perisinusoidal spaces (42.3 percent), as well as around the hepatic terminal vein (57.7 percent). Thus, hepatotrophic factors administration in the portal blood promoted a regenerative hepatic response, with an overall reduction of the volumetric density of collagen, improved hepatic function, and a general improvement in the histopathological aspects of fibrosis. CONCLUSION: Taken together, these results suggest the potential therapeutic use of this hepatotrophic factors solution to treat chronic liver diseases.

CONTEXTO: A fibrose hepática ocorre em resposta a diversos agentes agressores e é um fator predisponente da cirrose. Fatores hepatotróficos são conhecidos por estimular o crescimento hepático e restaurar a arquitetura histológica do fígado. Promovem, também, melhora na função hepática e aceleram a reversão da fibrose antes de sua progressão para cirrose. OBJETIVO: Testar os efeitos de uma solução de fatores hepatotróficos, composta por aminoácidos, vitaminas, glicose, insulina, glugacon e triiodotironina na fibrose hepática em ratos. MéTODOS: No presente estudo, a fibrose foi induzida em ratos pela administração de dimetilnitrosamina (10 mg/kg) durante 5 semanas. Após a biopsia do fígado, os ratos receberam a solução de fatores hepatotróficos (40 mg/kg/dia) ou solução salina por injeção intraperitonial, durante 10 dias. Amostras sanguíneas e fragmentos do fígado foram coletados para análise da função hepática, avaliação do critério histopatológico e quantificação morfométrica do colágeno. RESULTADOS: Os ratos do grupo fatores hepatotróficos demonstraram diminuição dos componentes histopatológicos da fibrose e aumento de massa hepática (12,2 por cento). Não houve o desenvolvimento de lesões neoplásicas em ambos os grupos. Comparado com o grupo de salina, no grupo fatores hepatotróficos também houve diminuição nos níveis dos marcadores sanguíneos de lesão hepática (AST e ALT), e diminuição da quantidade de colágeno nos espaços porta (31,6 por cento) e espaços perissinusoidais (42,3 por cento), assim como ao redor das veias terminais hepáticas (57,7 por cento). Assim, a administração de fatores hepatotróficos no sangue portal promoveu resposta regenerativa hepática, com redução da densidade volumétrica de colágeno, melhora na função hepática e melhora geral nos aspectos histopatológicos da fibrose. CONCLUSÃO: Juntos, estes resultados sugerem o potencial uso terapêutico desta solução de fatores hepatotróficos para tratar doenças hepáticas crônicas.

Reduction of liver mass due to malnutrition in rats: correlation with emaciation of animals and size of organs

We studied the effects of protein-energy malnutrition on the liver morphology of rats as compared to animal emaciation and to reduction in size of the organs not irrigated by splanchnic blood such as kidneys and spleen. The animals were divided into two groups, one of them fed ad libitum rate (N=10) and the other (N=14) receiving water but no food for 7 days, and the changes in animal weight, liver, kidney and spleen mass were determined. DNA and the protein/DNA ratio, as well as hepatocyte size, were determined in liver tissue. The liver decreased in mass (27.14 percent) at a significantly higher proportion (p<0.05) when compared to body emaciation (19.22 percent). Similar to the reduction in body weight, the masses of kidneys and spleen were reduced by 18.68 percent and 24.28 percent, respectively The reduction in liver mass occurred due to hypoplasia and atrophy, i.e., a decrease in hepatocyte number and size, respectively. We conclude that there is a preferential consumption of liver protein in protein-energy malnutrition which is suggested to result from the additive action of the effects of overall consumption of organic reserves due to malnutrition proper and to the reduction of the hepatotrophic stimulus.

Enhancement of liver size by stimulation of intact rat liver with exogenous hepatotrophic factors

Objetivos: Confirmar observaçoes anteriores (PARRA e col.) que sugeriam a possibilidade de fazer crescer o fígado em animais intactos, por estímulo hepatotrófico exógeno, ultrapassando seu tamanho biologicamente pré-determinado. Material e Métodos: Dois grupos de fêmeas de ratos Wistar, foram injetadas diariamente na cavidade peritoneal (40ml/kg) por sete dias consecutivos com as seguintes soluçoes: GRUPO A (controle) - soluçao salina com carboximetilcelulose (CMC) a 0,25 por cento; GRUPO B - soluçao de fatores hepatotróficos exógenos constituída de glicose, aminoácidos, insulina, glucagon, vitaminas, eletrólitos, triiodotironina e CMC na mesma concentraçao do grupo controle. No oitavo dia os animais forma sacrificados e avaliados os seguinte parâmetros: variaçao da massa hepática e DNA total do fígado dos animais do grupo experimental em relaçao ao grupo controle. Resultados: Foi observado crescimento da massa hepática de 114,16 +- 7,9 por cento acima do esperado e aumento do DNA total hepático de 12,99 +- 0,46 mg para 20,17 por fígado (p=0.0002). Conclusao: O aumento do tamanho do fígado a partir de sua massa primitiva, estimulada por fatores hepatotróficos exógenos, pode vir a ter aplicaçoes em intre-vivos, criando maior massa hepática a ser dividida entre doador e receptor.