Study of the integument that covering back and stinger of the freshwater stingray Potamotrygon rex (Chondricthyes, Potamotrygonidae).

The objective of this study was to describe the histology and histochemistry of the integument covering the back and stinger of the freshwater stingray Potamotrygon rex, endemic to the Middle Upper Tocantins River. The species has a dark back and yellowish circular spots that extend to the tail, which has one to two stings located in the median portion of the tail. Through histological study it was observed that the epithelia of the back and stinger are composed of non-keratinized stratified pavement epithelial tissue, and are organized in three layers: basal, intermediate and superficial. The differences between the tissues are related to the cell types that compose them. The back is organized with epithelial cells, mucus cells, granulocyte cells and chromatophores. The mucus cells are distributed in different layers along the animal's back, influencing the thickness of the tissue. The tissue that covers the stinger is composed of epithelial cells, chromatophores and specialized cells in protein synthesis. In the histochemistry, the stinger epithelial cells were stained with Bromophenol Blue, especially those of the intermediate layer, which were called specialized cells. In the back the epithelial cells were stained with Bromophenol Blue, Alcian Blue and PAS, and the mucous cells with PAS. In both tissues the presence of protein reserves was detected, areas so called because they are stained strongly with Bromophenol Blue. The results show that the stinger presents activity directed to the production of proteins, and that the back is organized to produce different components, which constitute the cuticle that covers the animal's body.

Study of the integument that covering back and stinger of the freshwater stingray Potamotrygon rex (Chondricthyes, Potamotrygonidae) / Estudo do tegumento que reveste o dorso e ferrão da arraia de água doce Potamotrygon rex (Chondricthyes, Potamotrygonidae)

The objective of this study was to describe the histology and histochemistry of the integument covering the back and stinger of the freshwater stingray Potamotrygon rex, endemic to the Middle Upper Tocantins River. The species has a dark back and yellowish circular spots that extend to the tail, which has one to two stings located in the median portion of the tail. Through histological study it was observed that the epithelia of the back and stinger are composed of non-keratinized stratified pavement epithelial tissue, and are organized in three layers: basal, intermediate and superficial. The differences between the tissues are related to the cell types that compose them. The back is organized with epithelial cells, mucus cells, granulocyte cells and chromatophores. The mucus cells are distributed in different layers along the animal's back, influencing the thickness of the tissue. The tissue that covers the stinger is composed of epithelial cells, chromatophores and specialized cells in protein synthesis. In the histochemistry, the stinger epithelial cells were stained with Bromophenol Blue, especially those of the intermediate layer, which were called specialized cells. In the back the epithelial cells were stained with Bromophenol Blue, Alcian Blue and PAS, and the mucous cells with PAS. In both tissues the presence of protein reserves was detected, areas so called because they are stained strongly with Bromophenol Blue. The results show that the stinger presents activity directed to the production of proteins, and that the back is organized to produce different components, which constitute the cuticle that covers the animal's body.

O objetivo deste estudo foi descrever a histologia e histoquímica do tegumento que reveste o dorso e o ferrão da arraia de água doce Potamotrygon rex, endêmica do Médio Alto Tocantins. A espécie possui o dorso escuro e manchas circulares amareladas que se estendem até a cauda, que possui de um a dois ferrões localizados na porção mediana da cauda. Através do estudo histológico observou-se que os epitélios do dorso e do ferrão são compostos por tecido epitelial pavimentoso estratificado não queratinizado, e estão organizados em três camadas: basal, intermediária e superficial. As diferenças entre os tecidos estão relacionadas aos tipos de células que os compõem. O dorso é organizado com células epiteliais, células mucosas, células de granulócitos e cromatóforos. As células mucosas estão distribuídas em diferentes camadas ao longo do dorso do animal, influenciando na espessura do tecido. O tecido que reveste o ferrão é composto por células epiteliais, cromatóforos e células proteicas. Na histoquímica, as células epiteliais do ferrão foram coradas com Azul de Bromofenol, principalmente as da camada intermediária, que foram denominadas células proteicas. No dorso as células epiteliais foram coradas com Azul de Bromofenol, Azul Alcian e PAS, e as células mucosas com PAS. Em ambos os tecidos também foi detectada a presença de reservatórios de proteínas, áreas assim denominadas por estarem fortemente coradas com Azul de Bromofenol. Os resultados mostram que o ferrão apresenta atividade direcionada à produção de proteínas, e que o dorso se organiza para produzir diferentes componentes que constituem a cutícula que reveste o corpo do animal.

Induction of haem oxygenase-1 increases infection of dog macrophages by L. infantum.

We aimed to induce and inhibit HO-1, ascertaining its effect on infection rate, parasite load and the levels of superoxide, reactive oxygen species (ROS), nitric oxide (NO), TNF-alpha and IL-10 in cultured macrophages from healthy dogs infected by Leishmania infantum. Macrophages obtained from 15 healthy dogs were cultured alone or infected with L. infantum, with or without association of HO-1 inducer and inhibitor. The infection rate and the parasite load were determined by the number of infected macrophages and number of promastigotes per macrophage, respectively. HO-1 levels and gene expression, as well as IL-10 and TNF-alpha levels were also measured in these cultures. Superoxide, ROS and NO levels in macrophages were measured through flow cytometry. Induction of HO-1 increased the infection rate and parasite load, while its inhibition decreased the infection rate and IL-10 production. There was a positive correlation between HO-1 and infection rate or parasite load. Increased infection rate was associated with decreased superoxide, ROS and NO levels. Induction of HO-1 metabolism in dogs infected by L. infantum is possibly one of the mechanisms responsible for increasing the infection of macrophages, mainly through reduction in the oxidative and nitrosative metabolisms of these cells.

Trajectories of attention-deficit/hyperactivity disorder dimensions in adults.

OBJECTIVE: There is a lack of available information on the trajectories of attention-deficit/hyperactivity disorder (ADHD) dimensions during adulthood. This study investigates the course and the predictors of change for each ADHD domain in a clinical sample of adults with ADHD. METHOD: Adults with ADHD (n = 344) were followed up for 7 years, with a final retention rate of 66.0%. Trajectories of inattention, hyperactivity, and impulsivity and their potential predictors were examined. RESULTS: On average, symptoms declined in all ADHD domains during follow-up. Despite this, rises in inattentive, hyperactive, and impulsive symptoms were observed in approximately 13%, 25%, and 17% of patients respectively. Different predictors influenced the trajectory of each ADHD dimension. Oppositional defiant disorder and social phobia were associated with the maintenance of symptoms, while alcohol use disorder was associated with both maintenance and rise of symptoms. CONCLUSION: Unexpectedly, a rise in the symptoms after 7 years was not uncommon in adults with ADHD. Prevalent comorbidities have the potential to influence the neurodevelopment and the trajectory of ADHD. Therefore, such predictors should be investigated in population cohorts to better characterize the course of ADHD. Additionally, these findings may be relevant in prevention studies and in strategies for ADHD treatment.

M1 polarization and the effect of PGE2 on TNF-α production by lymph node cells from dogs with visceral leishmaniasis.

Canine visceral leishmaniasis (CVL) is caused by the intracellular parasite Leishmania infantum. Increased levels of arginase, nitric oxide (NO2 ) and prostaglandin E2 (PGE2 ) can play a regulatory role regarding the immune response in CVL cases. This study aimed to evaluate the arginase activity in adherent macrophages cultured from the lymph nodes of healthy and naturally infected dogs and to examine the NO2 and PGE2 levels in the supernatant of these cultures. In addition, the regulatory effect of PGE2 on the production of tumour necrosis factor (TNF-α) and interleukin-10 (IL-10) in supernatants from the total lymph node was observed in leucocyte cultures. The arginase activity was lower in the adherent macrophages cultured from the lymph nodes of naturally infected dogs and there were higher concentrations of NO2 and PGE2 in the supernatants of these cultures. Higher TNF-α and IL-10 concentrations were observed in supernatants from total lymph node leucocytes cultures, from infected dogs, and the presence of indomethacin only decreased TNF-α in the supernatant of these cultures. We conclude that the low arginase activity in macrophages suggested that M1 polarization and PGE2 were participating in the immune response and were increasing TNF-α in CVL.

Recombinant K28 antigen in ELISA in the diagnosis of canine visceral leishmaniosis.

Crude total antigen (CTA) from Leishmania infantum and recombinant antigen K39 (rK39) and recombinant antigen K28 (rK28) were compared using an ELISA for the diagnosis of canine visceral leishmaniosis (CVL). Forty-two blood samples from healthy dogs from a nonendemic area and 80 blood samples from an endemic area for dogs with visceral leishmaniosis (VL), confirmed with positive parasitological tests for Leishmania spp., were used in an ELISA. The parasitological diagnosis was chosen as a gold standard. The ELISA with rK28 antigen showed sensitivity of 100% and specificity of 100%, high agreement with CTA and rK39, indicating that the rK28 antigen is useful for ELISA serological diagnosis of CVL.

Persistence and remission of ADHD during adulthood: a 7-year clinical follow-up study.

BACKGROUND: Course and predictors of persistence of attention deficit hyperactivity disorder (ADHD) in adults are still largely unknown. Neurobiological and clinical differences between child and adult ADHD raise the need for follow-up studies of patients diagnosed during adulthood. This study investigates predictors of ADHD persistence and the possibility of full remission 7 years after baseline assessment. METHOD: A 7-year follow-up study of adults with ADHD (n = 344, mean age 34.1 years, 49.9% males) was conducted. Variables from different domains (social demographics, co-morbidities, temperament, medication status, ADHD measures) were explored with the aim of finding potential predictors of ADHD persistence. RESULTS: Retention rate was 66% (n = 227). Approximately a third of the sample (n = 70, 30.2%) did not maintain ADHD criteria and 28 (12.4%) presented full remission (<4 symptoms), independently of changes in co-morbidity or cognitive demand profiles. Baseline predictors of diagnostic persistence were higher number of inattention symptoms [odds ratio (OR) 8.05, 95% confidence interval (CI) 2.54-25.45, p < 0.001], number of hyperactivity/impulsivity symptoms (OR 1.18, 95% CI 1.04-1.34, p = 0.01), oppositional defiant disorder (OR 3.12, 95% CI 1.20-8.11, p = 0.02), and social phobia (OR 3.59, 95% CI 1.12-11.47, p = 0.03). CONCLUSIONS: Despite the stage of brain maturation in adults suggests stability, approximately one third of the sample did not keep full DSM-IV diagnosis at follow-up, regardless if at early, middle or older adulthood. Although full remission is less common than in childhood, it should be considered as a possible outcome among adults.

Effects of P-MAPA immunomodulator on Toll-like receptor 2, ROS, nitric oxide, MAPKp38 and IKK in PBMC and macrophages from dogs with visceral leishmaniasis.

Leishmania (L.) chagasi is the etiologic agent of visceral leishmaniasis (VL) that can be transmitted to humans and dogs. VL in Brazil represents a serious public health problem; therefore, it is important to study new alternatives to treat infected dogs. In dogs, the therapeutic arsenal against canine VL is limited. The immunomodulator protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride (P-MAPA) improves immunocompetence when the immune system is impaired, but its dependence on Toll-like receptors (TLRs) and the mechanisms involved in immune response remain unclear. The in vitro action of P-MAPA on the expression of TLR2 and TLR4, reactive oxygen species (ROS), nitric oxide (NO) and p38 mitogen-activated protein kinase (p38 MAPK) and IKK phosphorylation was studied in mononuclear cells from peripheral blood and macrophages from healthy and Leishmania-infected dogs. The PBMC or macrophages were isolated and cultured with different concentrations of P-MAPA (20,100 and 200 µg/ml) in a humid environment at 37°C with 5% CO(2). Observation revealed that Leishmania-infected dogs showed a decrease in TLR2 in macrophages compared with healthy dogs and in induction with P-MAPA. ROS were increased in PBMCs from Leishmania spp.-infected dogs compared with healthy dogs and P-MAPA improved ROS production. NO production was increased in culture supernatant from macrophages stimulated by P-MAPA in both healthy and Leishmania spp. infected dogs. Treatment of macrophages from healthy dogs with immunomodulatory P-MAPA induced p38 MAPK and IKK phosphorylation, suggesting signal transduction by this pathway. These findings suggest that P-MAPA has potential as a therapeutic drug in the treatment of canine visceral leishmaniasis.

Desempenho de tilápias-do-nilo alimentadas com farelo da casca de pequi / Performance of Nile tilapia fed with bran made of pequi peel

Avaliou-se o desempenho de tilápias-do-nilo alimentadas com farelo da casca de pequi (Caryocar brasiliense). Foram utilizados 200 alevinos, com idade de 37 dias e peso corporal médio de 0,63±0,25g, distribuídos em delineamento inteiramente ao acaso, com quatro tratamentos ­ zero, 20, 40 e 60% de substituição de ração comercial por farelo da casca de pequi ­ e cinco repetições representadas por caixas de cloreto de polivinila com capacidade para 130L, contendo 10 peixes cada, totalizando 20 unidades experimentais. As características de desempenho avaliadas foram consumo de ração, peso corporal, ganho de peso, conversão alimentar, comprimento total e viabilidade criatória. A conversão alimentar ­ 1,96µ; 2,21µ; 2,63µ; 3,12µ - piorou linearmente com a inclusão do farelo de casca da pequi, enquanto as demais variáveis de desempenho não foram influenciadas pelos tratamentos. Conclui-se que a inclusão do farelo da casca de pequi na ração piora a conversão alimentar, sem alterar as demais variáveis de desempenho.

The performance of Nile tilapia fed with bran made of pequi peel was evaluated. Two hundred fingerlings, at 37 days of age and with mean body weight of 0.63±0.25 g, were distributed in a completely randomized design with four treatments ­0, 20, 40 and 60% of replacement of commercial diet with bran made of pequi peel ­with five repetitions represented by boxes of polyvinyl chloride (PVC) with capacity for 130 L, with 10 fish each, totalizing 20 experimental units. The performance characteristics evaluated were feed intake, body weight, weight gain, feed conversion, total length, and live viability. Feed conversion ­1.96µ; 2.21µ; 2.63µ; 3.12µ­ increased linearly with the inclusion of bran made of pequi peel, while the other performance variables were not influenced by treatments. The conclusion is that the inclusion of bran peel in the pequi diet worsened feed conversion, without changing other performance variables.

Desempenho de tilápias-do-nilo alimentadas com farelo da casca de pequi / Performance of Nile tilapia fed with bran made of pequi peel

Avaliou-se o desempenho de tilápias-do-nilo alimentadas com farelo da casca de pequi (Caryocar brasiliense). Foram utilizados 200 alevinos, com idade de 37 dias e peso corporal médio de 0,63±0,25g, distribuídos em delineamento inteiramente ao acaso, com quatro tratamentos ­ zero, 20, 40 e 60% de substituição de ração comercial por farelo da casca de pequi ­ e cinco repetições representadas por caixas de cloreto de polivinila com capacidade para 130L, contendo 10 peixes cada, totalizando 20 unidades experimentais. As características de desempenho avaliadas foram consumo de ração, peso corporal, ganho de peso, conversão alimentar, comprimento total e viabilidade criatória. A conversão alimentar ­ 1,96µ; 2,21µ; 2,63µ; 3,12µ - piorou linearmente com a inclusão do farelo de casca da pequi, enquanto as demais variáveis de desempenho não foram influenciadas pelos tratamentos. Conclui-se que a inclusão do farelo da casca de pequi na ração piora a conversão alimentar, sem alterar as demais variáveis de desempenho.(AU)

The performance of Nile tilapia fed with bran made of pequi peel was evaluated. Two hundred fingerlings, at 37 days of age and with mean body weight of 0.63±0.25 g, were distributed in a completely randomized design with four treatments ­0, 20, 40 and 60% of replacement of commercial diet with bran made of pequi peel ­with five repetitions represented by boxes of polyvinyl chloride (PVC) with capacity for 130 L, with 10 fish each, totalizing 20 experimental units. The performance characteristics evaluated were feed intake, body weight, weight gain, feed conversion, total length, and live viability. Feed conversion ­1.96µ; 2.21µ; 2.63µ; 3.12µ­ increased linearly with the inclusion of bran made of pequi peel, while the other performance variables were not influenced by treatments. The conclusion is that the inclusion of bran peel in the pequi diet worsened feed conversion, without changing other performance variables.(AU)

Fungos aeróbios no intestino grosso de borregos e de ovelhas criados em pastagens tropicais / Aerobic fungi in the large intestine of lambs and ewes raised on tropical pastures

Fungi populations were evaluated in large intestine from sheep raised in the North of Minas Gerais, Brazil. The samples were from 39 Santa Inês crossbred ewes and 30 lambs raised on Tanzania grass (Panicum maximum cv. Tanzania), both supplemented with mineral mixtures. The clinical specimens were directly collected from rectal ampoule with sterile swabs. The development of fungal mycelium was observed in all lamb samples and positive cultures were observed in 34 ewe specimens (87.2 percent). After microculture of 40 colonies from lambs, 34 isolates of the genus Aspergillus, three of Paecilomyces spp., one Acremonium sp., and one Trichoderma sp. were identified. Out of the 39 isolates from ewes, 15 Paecilomyces spp., 11 Aspergillus spp., 11 Malbranchea spp., and one Onychocola sp. were identified. The results showed the predominance of the genus Aspergillus in the lambs, while the genus Paecilomyces, Malbranchea, and Aspergillus were predominant, in ewes.(AU)

Fungos aeróbios no intestino grosso de borregos e de ovelhas criados em pastagens tropicais / Aerobic fungi in the large intestine of lambs and ewes raised on tropical pastures

Fungi populations were evaluated in large intestine from sheep raised in the North of Minas Gerais, Brazil. The samples were from 39 Santa Inês crossbred ewes and 30 lambs raised on Tanzania grass (Panicum maximum cv. Tanzania), both supplemented with mineral mixtures. The clinical specimens were directly collected from rectal ampoule with sterile swabs. The development of fungal mycelium was observed in all lamb samples and positive cultures were observed in 34 ewe specimens (87.2 percent). After microculture of 40 colonies from lambs, 34 isolates of the genus Aspergillus, three of Paecilomyces spp., one Acremonium sp., and one Trichoderma sp. were identified. Out of the 39 isolates from ewes, 15 Paecilomyces spp., 11 Aspergillus spp., 11 Malbranchea spp., and one Onychocola sp. were identified. The results showed the predominance of the genus Aspergillus in the lambs, while the genus Paecilomyces, Malbranchea, and Aspergillus were predominant, in ewes.

Distribution of human immunodeficiency virus type 1 subtypes in the state of Amazonas, Brazil, and subtype C identification

Few studies have reported the molecular epidemiological characterization of HIV-1 in the Northern region of Brazil. The present study reports the molecular and epidemiological characterization of 31 HIV-1 isolates from blood donors from the State of Amazonas who donated blood between April 2006 and March 2007. Serum/plasma samples from all donors were screened for HIV antibodies by ELISA and the results confirmed by Western blot analysis. Genomic DNA was extracted from the buffy coat using the Super Quik-Gene-DNA Isolation kit. Nested PCR was performed on the env, gag, and pol regions of HIV-1 using the Gene Amp PCR System 9700. Sequencing reactions were performed using the inner PCR primers and the DYEnamic™ ET Dye Terminator Kit, and phylogenetic analysis was performed using the gag, pol, and env gene sequences. We collected samples from 31 blood donors who tested positive for HIV-1 in confirmatory experiments. The male:female ratio of blood donors was 3.4:1, and the mean age was 32.4 years (range: 19 to 61 years). Phylogenetic analysis showed that subtype B is the most prevalent among Northern Brazilian HIV-1-seropositive blood donors. One HIV-1 subtype C and one circulating recombinant form (CRF_BF) of HIV-1 were identified in the State of Amazonas. This is the first study showing the occurrence of a possible "homogenous" subtype C in this region of Brazil. This finding could contribute to a better characterization of the HIV-1 strains that circulate in the country.

Distribution of human immunodeficiency virus type 1 subtypes in the State of Amazonas, Brazil, and subtype C identification.

Few studies have reported the molecular epidemiological characterization of HIV-1 in the Northern region of Brazil. The present study reports the molecular and epidemiological characterization of 31 HIV-1 isolates from blood donors from the State of Amazonas who donated blood between April 2006 and March 2007. Serum/plasma samples from all donors were screened for HIV antibodies by ELISA and the results confirmed by Western blot analysis. Genomic DNA was extracted from the buffy coat using the Super Quik-Gene-DNA Isolation kit. Nested PCR was performed on the env, gag, and pol regions of HIV-1 using the Gene Amp PCR System 9700. Sequencing reactions were performed using the inner PCR primers and the DYEnamic™ ET Dye Terminator Kit, and phylogenetic analysis was performed using the gag, pol, and env gene sequences. We collected samples from 31 blood donors who tested positive for HIV-1 in confirmatory experiments. The male:female ratio of blood donors was 3.4:1, and the mean age was 32.4 years (range: 19 to 61 years). Phylogenetic analysis showed that subtype B is the most prevalent among Northern Brazilian HIV-1-seropositive blood donors. One HIV-1 subtype C and one circulating recombinant form (CRF_BF) of HIV-1 were identified in the State of Amazonas. This is the first study showing the occurrence of a possible "homogenous" subtype C in this region of Brazil. This finding could contribute to a better characterization of the HIV-1 strains that circulate in the country.

The burdened life of adults with ADHD: impairment beyond comorbidity.

Since approximately 70% of adult patients with attention-deficit/hyperactivity disorder (ADHD) have at least one comorbid disorder, rating of impairment specifically attributable to ADHD is a hard task. Despite the evidence linking environmental adversities with negative outcomes in ADHD, life events measures have not been used to rate the disorder impairment. The present study tested for the first time the hypothesis that increased ADHD severity is associated with an increase in negative recent life events, independently of comorbidity status. The psychiatric diagnoses of 211 adult ADHD outpatients were based on DSM-IV criteria assessed through structured interviews (K-SADS-E for ADHD and ODD, MINI for ASPD and SCID-IV-R for other comorbidities). ADHD severity was evaluated with the Swanson, Nolan and Pelham rating scale (SNAP-IV) and recent life events with the Life Experience Survey. Higher SNAP-IV inattention and hyperactivity scores, female gender, lower socioeconomic status and the presence of comorbid mood disorders were associated with negative life events. Poisson regression models with adjustment for possible confounders confirmed the effect of inattention and hyperactivity severity on negative life events. Our results suggest that the negative life events experienced by these patients are associated to the severity of ADHD independently from comorbid psychiatric disorders.

Apoptotic effect of fludarabine is independent of expression of IAPs in B-cell chronic lymphocytic leukemia.

Despite the efficiency of fludarabine in the induction of clinical responses in B-cell chronic lymphocytic leukemia (B-CLL) patients, resistance to this drug has been documented. The present study tested whether resistance to fludarabine is related to the expression of inhibitor of apoptosis proteins (IAPs) family members. We analyzed the expression of c-IAP1, c-IAP2 and XIAP, by immunocytochemistry, in 30 blood samples from B-CLL patients and correlated protein expression to fludarabine-induced apoptosis estimated by an annexin-V assay. Expression of c-IAP1, c-IAP2 and XIAP were found predominantly in the cytoplasm, and a wide range of staining intensities was observed among distinct samples. No correlation was found between the levels of IAPs expression and prognostic factors such as age, gender, lymphocyte doubling time, white blood cell count or previous treatment. The expression of IAPs also failed to predict the sensitivity to fludarabine-induced apoptosis. Alternative pathways of cell death may explain the independence of fludarabine-induced apoptosis from the high expression of IAPs.

Chemical composition and analgesic activity of Calophyllum brasiliense leaves.

This paper describes a phytochemical and pharmacological study with Calophyllum brasiliense leaves, a medicinal plant employed in folk medicine for the treatment of several ailments. Based on spectroscopic evidence, five phenolic compounds were identified as hyperin (hyperoside), amentoflavone, quercetin, gallic acid, and protocatechuic acid. The fractions and some phenolic compounds exhibited significant analgesic activity against the writhing test and in relation to the second phase (inflammatory pain) of the formalin test in mice, suggesting that this plant can be useful for the treatment of dolorous processes.

I3-naringenin-II8--4'OMe-eriodictyol: a new potential analgesic agent isolated from Rheedia gardneriana leaves.

This paper describes the isolation, identification and analgesic activity of a new biflavonoid from Rheedia gardneriana leaves, which correspond to I3-naringenin-II8-4'-OMe-eriodictyol (GB-2a-II-4'-OMe) (1), with a methoxyl group in position 4 of ring-II. Its structure was determined by spectroscopic data and confirmed by an alkaline hydrolysis. Its analgesic effect was evaluated in a writhing test and a formalin test in mice. It was found that this compound exhibits potent and dose-related analgesic action in both experimental models, with ID50's values of 4.5 micromol/kg against the writhing test and 8.2 and 6.8 micromol/kg against the first and second phase of the formalin test, respectively. It was several times more potent than some well-known analgesic drugs used as reference.

Characterization and distribution of extracellular matrix components and receptors in GH3B6 prolactin cells.

GH3B6 cells, a rat pituitary tumor cell line, synthesize and secrete large amounts of prolactin (PRL) in vitro. In the present work, we evaluated the capacity of these cells to express extracellular matrix (ECM) components and receptors in vitro. The expression of laminin (LN), fibronectin (FN) and type IV collagen (CIV) was investigated by immunofluorescence assays. In comparison to PRL distribution, where around 50-70% of the cells contained PRL concentrated in the Golgi region, a variable immunolabeling for the three ECM components could be observed in the majority of GH3B6 cells. Importantly, this pattern was not modified when cells were cultured in the presence of 30 nM thyroliberin (TRH). The expression of the ECM receptors: alpha5beta1 (FN receptor), alpha6beta1 (LN receptor) and CD44 (hyaluronic acid receptor) could be demonstrated by cytofluorometric analysis. Using biochemical procedures, we analyzed the synthesis and secretion of glycosaminoglycans (GAGs). The cells synthesized and secreted mainly heparan sulfate (75%) with a minor amount of chondroitin sulfate/dermatan sulfate. In an attempt to evaluate the individual contribution of the ECM components to influence cell morphology and PRL distribution in vitro, GH3B6 cells were cultivated separately on LN, FN and CIV substrates. Under all conditions, it was possible to observe an increase of cell adherence to the substrate, accompanied with changes of cellular morphology, characterized by the appearance of cytoplasmatic processes. However, no changes on PRL distribution could be observed. Our results suggest that endocrine tumor cell lines are involved in synthesis of ECM components and receptors.