RESUMO
Diclofenac is the most prescribed nonsteroidal anti-inflammatory drug worldwide and is used to relieve pain and inflammation in inflammatory arthritis. Diclofenac is associated with serious adverse effects, even in regular-dose regimens. Drug delivery systems can overcome this issue by reducing adverse effects and optimizing their efficacy. This study evaluated the activity of lipid-core nanocapsules loaded with diclofenac (DIC-LNCs) in an experimental model of adjuvant-induced arthritis. The diclofenac nanoformulation was obtained via self-assembly. A stereological analysis approach was applied for the morphological quantification of the volume, density, and cellular profile count of the metatarsophalangeal joints of rats. Proinflammatory cytokines and biochemical profiles were also obtained. Our results showed that the diclofenac nanocapsule DIC-LNCs were able to reduce arthritis compared with the control group and the DIC group. DIC-LNCs efficiently reduced proinflammatory cytokines, C-reactive protein, and xanthine oxidase levels. Additionally, DIC-LNCs reduced the loss of synoviocytes and chondrocytes compared with the DIC (p < 0.05) and control groups (p < 0.05). These data suggest that DIC-LNCs have anti-arthritic activity and preserve joint components, making them promising for clinical use.
Assuntos
Artrite Experimental , Nanocápsulas , Ratos , Animais , Diclofenaco/farmacologia , Diclofenaco/uso terapêutico , Artrite Experimental/tratamento farmacológico , Lipídeos/uso terapêutico , CitocinasRESUMO
The egg is one of the fundamental parts of the life cycle of Neoechinorhynchus buttnerae, and this stage involves the acanthor larva. It is also the infection phase for the intermediate host. Under normal conditions, the larva inside the egg can survive for months in the environment; however, information regarding this phase of life of the parasite is scarce. In addition, there is no quantitative information about the structural composition of the parasite's body from a histological point of view. Such information is essential in order to support decisions aimed at controlling infestations by these parasites in fish farming. This study aimed to present a detailed description of the stages of embryonic development of N. buttnerae eggs, as well as a stereological evaluation of the body of adult females of the parasite. Three phases of development characterized the eggs: cell division (with four stages), formation of the internal nuclear mass (with four stages) and formation of the acanthor larva (with five stages). The ovary comprised 26.61% of the volume of the animal and most of it contained eggs (21.28%), ovarian balls (3.88%) and empty spaces (1.45%). These results are of great importance and will support future studies that seek to interrupt the life cycle of this parasite.
Assuntos
Acantocéfalos , Doenças dos Peixes , Helmintíase Animal , Animais , Desenvolvimento Embrionário , Feminino , Doenças dos Peixes/parasitologia , Helmintíase Animal/parasitologia , Larva , Estágios do Ciclo de VidaRESUMO
Numerous mechanisms have been proposed to explain why patients with malaria are more susceptible to bloodstream invasions by Salmonella spp., however there are still several unknown critical factors regarding the pathogenesis of coinfection. From a coinfection model, in which an S. enterica serovar Typhi (S_Typhi) was chosen to challenge mice that had been infected 24 h earlier with Plasmodium berghei ANKA (P.b_ANKA), we evaluated the influence of malaria on cytokine levels, the functional activity of femoral bone marrow-derived macrophages and neutrophils, and intestinal permeability. The cytokine profile over eight days of coinfection showed exacerbation in the cytokines MCP-1, IFNγ and TNFα in relation to the increase seen in animals with malaria. The cytokine profile was associated with a considerably reduced neutrophil and macrophage count and a prominent dysfunction, especially in ex vivo neutrophils in coinfected mice, though without bacterial modulation that could influence the invasion capacity of ex vivo S_Typhi obtained from liver macerate in non-phagocyte cells. Finally, irregularities in the integrity of intestinal tissue evidenced ruptures in the enterocyte layer, a presence of mononuclear leukocytes in the enterocyte layer, an increase of goblet cells in the enterocyte layer and a high volume of leukocyte infiltrate in the sub-mucosa were greatly increased in coinfected animals. Increases of mononuclear leukocytes in the enterocyte layer and volume of leukocyte infiltrate in the sub-mucosa were also seen in monoinfected animals with P. berghei ANKA. Our findings suggest malaria causes a disarrangement of intestinal homeostasis, exacerbation of proinflammatory cytokines and dysfunction in neutrophils that render the host susceptible to bacteremia by Salmonella spp.
Assuntos
Fígado/patologia , Malária/patologia , Febre Tifoide/patologia , Animais , Coinfecção/patologia , Modelos Animais de Doenças , Macrófagos/patologia , Camundongos , Camundongos Endogâmicos BALB C , Neutrófilos/patologia , Plasmodium berghei , Salmonella typhiRESUMO
The aim of this study was to evaluate the impacts of metazoan parasites on hematological and biochemical parameters and relative condition factor of tambaqui (Colossoma macropomum ) farmed in northern Brazil. A total of 32 juvenile fish were captured from a commercial fish farm located in the municipality of Rio Preto da Eva, Amazonas state, Brazil. Parasite prevalence was 100% for Anacanthorus spathulatus, Mymarothecium boegeri and Notozothecium janauachensis, 100% for Neoechinorhynchus buttnerae and 53.13% for Dolops geayi. The greatest mean parasite intensity was found in acantocephalans followed by monogeneans and branchiuran crustaceans. A negative correlation was observed between abundance of N. buttnerae and hematocrit percentage, hemoglobin concentration, total thrombocyte count and glucose and between abundance of the monogenean and glucose concentration. Parasitic infections caused damage in tambaqui in terms of the observed hematological parameters that were characterized by hypochromic anemia and thrombocytopenia, which are important parameters to be used in parasitic diagnosis. This study is the first record of the occurrence of Dolops geayi in farmed tambaqui in the Amazon.(AU)
O objetivo deste estudo foi avaliar o impacto do parasitismo por metazoários sobre os parâmetros hematológicos, bioquímicos e fator de condição relativo de tambaquis (Colossoma macropomum) cultivados na região Norte do Brasil. Foram coletados 32 tambaquis em fase de engorda numa piscicultura comercial no município de Rio Preto da Eva, estado do Amazonas, Brasil. A prevalência de parasitos foi de 100% para Anacanthorus spathulatus, Mymarothecium boegeri e Notozothecium janauachensis, 100% para Neoechinorhynchus buttnerae e 53,13% para Dolops geayi. A maior intensidade média foi de acantocéfalos, seguida por monogenéticos e branquiúros. Correlação negativa significativa foi observada entre abundância de N. buttnerae e os parâmetros de hematócrito, concentração de hemoglobina, trombócitos totais e glicose, bem como entre a abundância de monogenéticos e glicose. A infecção por parasitos metazoários foi capaz de debilitar os tambaquis como observado por meio das alterações hematológicas; sendo este quadro de anemia hipocrômica e trombocitopenia importante para ser utilizado no diagnóstico destas parasitoses. Este estudo foi o primeiro registro da ocorrência de Dolops geayi em tambaquis cultivados na Amazônia.(AU)
Assuntos
Animais , Characidae/sangue , Characidae/parasitologia , Parasitos , Acantocéfalos , Anelídeos , Pesqueiros , BrasilRESUMO
Abstract The aim of this study was to evaluate the impacts of metazoan parasites on hematological and biochemical parameters and relative condition factor of tambaqui (Colossoma macropomum ) farmed in northern Brazil. A total of 32 juvenile fish were captured from a commercial fish farm located in the municipality of Rio Preto da Eva, Amazonas state, Brazil. Parasite prevalence was 100% for Anacanthorus spathulatus, Mymarothecium boegeri and Notozothecium janauachensis, 100% for Neoechinorhynchus buttnerae and 53.13% for Dolops geayi. The greatest mean parasite intensity was found in acantocephalans followed by monogeneans and branchiuran crustaceans. A negative correlation was observed between abundance of N. buttnerae and hematocrit percentage, hemoglobin concentration, total thrombocyte count and glucose and between abundance of the monogenean and glucose concentration. Parasitic infections caused damage in tambaqui in terms of the observed hematological parameters that were characterized by hypochromic anemia and thrombocytopenia, which are important parameters to be used in parasitic diagnosis. This study is the first record of the occurrence of Dolops geayi in farmed tambaqui in the Amazon.
Resumo O objetivo deste estudo foi avaliar o impacto do parasitismo por metazoários sobre os parâmetros hematológicos, bioquímicos e fator de condição relativo de tambaquis (Colossoma macropomum) cultivados na região Norte do Brasil. Foram coletados 32 tambaquis em fase de engorda numa piscicultura comercial no município de Rio Preto da Eva, estado do Amazonas, Brasil. A prevalência de parasitos foi de 100% para Anacanthorus spathulatus, Mymarothecium boegeri e Notozothecium janauachensis, 100% para Neoechinorhynchus buttnerae e 53,13% para Dolops geayi. A maior intensidade média foi de acantocéfalos, seguida por monogenéticos e branquiúros. Correlação negativa significativa foi observada entre abundância de N. buttnerae e os parâmetros de hematócrito, concentração de hemoglobina, trombócitos totais e glicose, bem como entre a abundância de monogenéticos e glicose. A infecção por parasitos metazoários foi capaz de debilitar os tambaquis como observado por meio das alterações hematológicas; sendo este quadro de anemia hipocrômica e trombocitopenia importante para ser utilizado no diagnóstico destas parasitoses. Este estudo foi o primeiro registro da ocorrência de Dolops geayi em tambaquis cultivados na Amazônia.
Assuntos
Animais , Arguloida , Caraciformes/parasitologia , Doenças dos Peixes/parasitologia , Helmintíase Animal/parasitologia , Helmintos/classificação , Aquicultura , Caraciformes/sangue , Doenças dos Peixes/sangue , Helmintíase Animal/sangueRESUMO
The aim of this study was to evaluate the impacts of metazoan parasites on hematological and biochemical parameters and relative condition factor of tambaqui (Colossoma macropomum ) farmed in northern Brazil. A total of 32 juvenile fish were captured from a commercial fish farm located in the municipality of Rio Preto da Eva, Amazonas state, Brazil. Parasite prevalence was 100% for Anacanthorus spathulatus, Mymarothecium boegeri and Notozothecium janauachensis, 100% for Neoechinorhynchus buttnerae and 53.13% for Dolops geayi. The greatest mean parasite intensity was found in acantocephalans followed by monogeneans and branchiuran crustaceans. A negative correlation was observed between abundance of N. buttnerae and hematocrit percentage, hemoglobin concentration, total thrombocyte count and glucose and between abundance of the monogenean and glucose concentration. Parasitic infections caused damage in tambaqui in terms of the observed hematological parameters that were characterized by hypochromic anemia and thrombocytopenia, which are important parameters to be used in parasitic diagnosis. This study is the first record of the occurrence of Dolops geayi in farmed tambaqui in the Amazon.
Assuntos
Arguloida , Caraciformes/parasitologia , Doenças dos Peixes/parasitologia , Helmintíase Animal/parasitologia , Helmintos/classificação , Animais , Aquicultura , Caraciformes/sangue , Doenças dos Peixes/sangue , Helmintíase Animal/sangueRESUMO
ABSTRACT The gill mitochondria-rich cells of the juvenile Amazonian fish Colossoma macropomum were analyzed using light and scanning and transmission electron microscopy after 96 h exposure to 0.04 and 0.2 mM nitrite. Although the number of mitochondria-rich cells decreased significantly in the lamellar epithelium, no decrease was found in the interlamellar region of the gill filament. Nitrite exposure caused significant reduction on the apical surface area of individual mitochondria-rich cells (p < 0.05), with a resulting reduction of the fractional area of these cells in both the lamellar and filament epithelium. Swelling of endoplasmic reticulum cisternae, nuclear envelope and mitochondria were the main changes found in the mitochondria-rich cells. Cristae lysis and matrix vacuolization characterized the mitochondrial changes. The overall ultrastructural changes indicated cellular functional disruption caused by exposure to nitrite. The changes observed in the gill indicate that the cellular structures involved in the process of energy production become severely damaged by exposure to nitrite indicating irreversible damage conducting to cell death.
Assuntos
Animais , Morte Celular , Exposição Ambiental , Characidae , Brânquias/citologia , Brânquias/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Análise de Variância , Microscopia Eletrônica de Transmissão , Nitritos/toxicidadeRESUMO
The gill mitochondria-rich cells of the juvenile Amazonian fish Colossoma macropomum were analyzed using light and scanning and transmission electron microscopy after 96 h exposure to 0.04 and 0.2 mM nitrite. Although the number of mitochondria-rich cells decreased significantly in the lamellar epithelium, no decrease was found in the interlamellar region of the gill filament. Nitrite exposure caused significant reduction on the apical surface area of individual mitochondria-rich cells (p < 0.05), with a resulting reduction of the fractional area of these cells in both the lamellar and filament epithelium. Swelling of endoplasmic reticulum cisternae, nuclear envelope and mitochondria were the main changes found in the mitochondria-rich cells. Cristae lysis and matrix vacuolization characterized the mitochondrial changes. The overall ultrastructural changes indicated cellular functional disruption caused by exposure to nitrite. The changes observed in the gill indicate that the cellular structures involved in the process of energy production become severely damaged by exposure to nitrite indicating irreversible damage conducting to cell death.
Assuntos
Morte Celular , Characidae , Exposição Ambiental , Brânquias/citologia , Brânquias/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Nitritos/toxicidade , Análise de Variância , Animais , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de TransmissãoRESUMO
The reproductive biology of South American freshwater stingrays (family Potamotrygonidae) is still poorly studied compared to other marine species. In the present study, we examined the gross anatomy and histology of six species of potamotrygonids from the Amazon basin and described the structural asymmetry of the ovaries and their relationship between ovarian and uterine fecundities. Stereological techniques were used to quantify the volume of ovarian and epigonal organ tissue associated with the left and right sides of the Potamotrygon wallacei, a recently described species, locally known as the cururu ray. This species presented ovarian asymmetry; the left epigonal organ-ovary complex was 55 times larger than the right side. The right side was composed of, volumetrically, 7.3% ovarian tissue and 92.7% epigonal organ tissue whereas the left side was 51.2% of ovarian tissue and 48.8% epigonal organ tissue. In all species, six types of follicles were identified in both right and left ovaries. Uteri were symmetrical and the fecundity ratio between the right and left sides was 0.9:1.1, respectively. Despite the volumetric difference of ovarian tissue between the two sides, the uterine fecundity shows that both ovaries are functional and that ovarian fecundity alone is not an accurate measure to determine the reproductive potential of freshwater stingrays. Anat Rec, 300:265-276, 2017. © 2016 Wiley Periodicals, Inc.
Assuntos
Ovário/anatomia & histologia , Rajidae/anatomia & histologia , Útero/anatomia & histologia , Animais , Feminino , Fertilidade/fisiologia , Folículo Ovariano/anatomia & histologia , Folículo Ovariano/fisiologia , Ovário/fisiologia , Reprodução/fisiologia , Rajidae/fisiologia , Útero/fisiologiaRESUMO
Injuries caused by freshwater stingrays are characterized by intense pain and pathological changes at the lesion site, including oedema, erythema and, in most cases, necrosis. In this study, the systemic myotoxic activity induced by mucus extracts from the dorsal region and stinger of the stingrays Plesiotrygon iwamae and Potamotrygon motoro was described, analysed and quantified. Twenty-four hours after injection of 400 µg of the extracts into the gastrocnemius muscle of mice, the following effects were observed: coagulative necrosis of the muscle tissue, muscle fibre regeneration and the presence of inflammatory infiltrates, including neutrophils, macrophages, and a reduced number of eosinophils and lymphocytes. These changes were also observed, although to a lesser extent, in the gastrocnemius muscles of the contralateral limbs, demonstrating that the extracts from the two species could induce systemic rhabdomyolysis. Based on morphometric analysis, it was observed that the stinger extract of P. motoro was more potent in inducing local and systemic myotoxic activity, followed by the dorsal extract from P. motoro and stinger and dorsal extracts from P. iwamae, which induced similar effects.
Assuntos
Mordeduras e Picadas/patologia , Venenos de Peixe/toxicidade , Muco/química , Rabdomiólise/induzido quimicamente , Rajidae/metabolismo , Animais , Mordeduras e Picadas/complicações , Brasil , Venenos de Peixe/análise , Água Doce , Macrófagos/patologia , Camundongos , Músculo Esquelético/patologia , Neutrófilos/patologia , Rabdomiólise/patologia , Especificidade da EspécieRESUMO
The gill structure of the Amazonian fish Arapaima gigas, an obligatory air breather, was investigated during its transition from water breathing to the obligatory air breathing modes of respiration. The gill structure of A. gigas larvae is similar to that of most teleost fish; however, the morphology of the gills changes as the fish grow. The main morphological changes in the gill structure of a growing fish include the following: (1) intense cell proliferation in the filaments and lamellae, resulting in increasing epithelial thickness and decreasing interlamellar distance; (2) pillar cell system atrophy, which reduces the blood circulation through the lamellae; (3) the generation of long cytoplasmic processes from the epithelial cells into the intercellular space, resulting in continuous and sinuous paracellular channels between the epithelial cells of the filament and lamella that may be involved in gas, ion, and nutrient transport to epithelial cells; and (4) intense mitochondria-rich cell (MRC) proliferation in the lamellar epithelium. All of these morphological changes in the gills contribute to a low increase of the respiratory surface area for gas exchange and an increase in the water-blood diffusion distance increasing their dependence on air-breathing as fish developed. The increased proliferation of MRCs may contribute to increased ion uptake, which favors the regulation of ion content and pH equilibrium.
Assuntos
Aclimatação/fisiologia , Células Epiteliais/citologia , Peixes/anatomia & histologia , Brânquias/anatomia & histologia , Mitocôndrias/fisiologia , Osmose/fisiologia , Respiração , Animais , Apoptose , Transporte Biológico , Proliferação de Células , Células Epiteliais/fisiologia , Peixes/fisiologia , Brânquias/fisiologia , Técnicas Imunoenzimáticas , Microscopia EletrônicaRESUMO
OBJECTIVES: To assess the economic impact of a 100% smoke-free law on bars and restaurants in an Argentinean province. MATERIALS AND METHODS: We conducted a time series analysis of restaurant and bar revenues in the province of Santa Fe 31 months before and 29 months after the implementation of the 100% smoke-free environment law. The neighboring province of Entre Rios without smoking restrictions at the time of this study, was used as the control province. RESULTS: Average taxable revenues post-legislation in the province of Santa Fe as a whole and in the two most important cities were higher when compared to the total provincial revenue pre-legislation. No significant differences were observed with the total revenue from the province of Entre Rios. CONCLUSIONS: We found no statistically significant evidence that the 100% smoke-free environment legislation in the province of Santa Fe, Argentina, had a negative impact on the revenues of local bars and restaurants.
Assuntos
Restaurantes/legislação & jurisprudência , Poluição por Fumaça de Tabaco/prevenção & controle , Argentina , Combustíveis Fósseis/estatística & dados numéricos , Humanos , Política Pública , Restaurantes/economia , Salários e Benefícios/estatística & dados numéricos , Estações do Ano , Impostos/estatística & dados numéricos , Poluição por Fumaça de Tabaco/legislação & jurisprudênciaRESUMO
OBJECTIVES: To assess the economic impact of a 100% smoke-free law on bars and restaurants in an Argentinean province. MATERIALS AND METHODS: We conducted a time series analysis of restaurant and bar revenues in the province of Santa Fe 31 months before and 29 months after the implementation of the 100% smokefree environment law. The neighboring province of Entre Rios without smoking restrictions at the time of this study, was used as the control province. RESULTS: Average taxable revenues post-legislation in the province of Santa Fe as a whole and in the two most important cities were higher when compared to the total provincial revenue pre-legislation. No significant differences were observed with the total revenue from the province of Entre Rios. CONCLUSIONS: We found no statistically significant evidence that the 100% smoke-free environment legislation in the province of Santa Fe, Argentina, had a negative impact on the revenues of local bars and restaurants.
OBJETIVOS: Evaluar el impacto económico de una ley de ambientes 100% libres de humo de tabaco sobre las ventas de bares y restaurantes en una provincia argentina. MATERIAL y MÉTODOS: Se realizó un análisis de serie temporal sobre las ventas de bares y restaurantes en Santa Fe, 31 meses antes y 29 meses después de la ley. Se utilizó a la provincia vecina de Entre Ríos, sin ley en el momento del estudio, como provincia control. RESULTADOS: El promedio de ventas post-ley tanto en la provincia de Santa Fe como en sus dos ciudades más importantes fue mayor al compararse con el promedio provincial total pre-ley. No se observaron diferencias significativas en las ventas totales al ser comparadas con la provincia de Entre Ríos. CONCLUSIONES: No se demostró evidencia estadísticamente significativa sobre el impacto negativo de la legislación de ambientes 100% libres de humo sobre las ventas de bares y restaurantes en la provincia de Santa Fe.
Assuntos
Humanos , Restaurantes/legislação & jurisprudência , Poluição por Fumaça de Tabaco/prevenção & controle , Argentina , Combustíveis Fósseis , Política Pública , Restaurantes/economia , Salários e Benefícios/estatística & dados numéricos , Estações do Ano , Impostos/estatística & dados numéricos , Poluição por Fumaça de Tabaco/legislação & jurisprudênciaRESUMO
The gills and the respiratory swim bladders of juvenile specimens (mean body mass 100g) of the basal teleost Arapaima gigas (Cuvier 1829) were evaluated using stereological methods in vertical sections. The surface areas, harmonic mean barrier thicknesses and morphometric diffusing capacities for oxygen and carbon dioxide were estimated. The average respiratory surface area of the swim bladder (2173 cm² kg⻹) exceeded that of the gills (780 cm² kg⻹) by a factor of 2.79. Due to the extremely thin air-blood barrier in the swim bladder (harmonic mean 0.22 µm) and the much thicker water-blood barrier of the gills (9.61 µm), the morphometric diffusing capacity for oxygen and carbon dioxide was 88 times greater in the swim bladder than in the gills. These data clearly indicate the importance of the swim bladder, even in juvenile A. gigas that still engage in aquatic respiration. Because of the much greater diffusion constant of CO2 than O2 in water, the gills also remain important for CO2 release.
Assuntos
Sacos Aéreos , Peixes , Brânquias , Consumo de Oxigênio , Sacos Aéreos/anatomia & histologia , Sacos Aéreos/fisiologia , Sacos Aéreos/ultraestrutura , Animais , Difusão , Peixes/anatomia & histologia , Peixes/fisiologia , Brânquias/anatomia & histologia , Brânquias/fisiologia , Brânquias/ultraestrutura , Microscopia/métodos , Microscopia Eletrônica de Transmissão , RespiraçãoRESUMO
PURPOSE: To morphometrically quantify CD1a+ dentritic cells and DC-SIGN+ dendritic cells in HIV-positive patients with anal squamous intraepithelial neoplasia and to evaluate the effects of HIV infection, antiretroviral therapy and HPV infection on epithelial and subepithelial dendritic cells. METHODS: A prospective study was performed to morphometrically analyze the relative volume of the dendritic cells and the relationship between anal intraepithelial neoplasia and cancer in HIV-positive patients from the Tropical Medicine Foundation of Amazonas, Brazil. All patients were submitted to biopsies of anorectal mucosa to perform a classic histopathological and immunohistochemical analysis, employing antibodies against CD1a and DC-SIGN for the morphometric quantification of dendritic cells. RESULTS: HIV-negative patients displayed a CD1a DC density significantly higher than that of HIV-positives patients (3.75 versus 2.54) (p=0.018), and in patients with severe anal intraepithelial neoplasia had correlated between DC CD1a density with levels of CD4 + cells (p: 0.04) as well as the viral load of HIV-1 (p: 0.035). A not significant rise in the median density of CD1a+ DC was observed in the HIV positive/ HAART positive subgroup compared to the HIV positive/ HAART negative subgroup. The CD1a+ DC were also significantly increased in HIV-negative patients with anorectal condyloma (2.33 to 3.53; p=0.05), with an opposite effect in HIV-positive patients. CONCLUSIONS: Our data support an enhancement of the synergistic action caused by HIV-HPV co-infection on the anal epithelium, weakening the DC for its major role in immune surveillance. Notoriously in patients with severe anal intraepithelial neoplasia, the density of CD1a+ epithelial dendritic cells was influenced by the viral load of HIV-1.(AU)
OBJETIVO: Quantificar morfometricamente as células dendríticas DC CD1a+ e DC DC-SIGN+ em pacientes HIV positivos portadores de neoplasia escamosa intraepitelial anal e avaliar os efeitos da infecção pelo HIV, da terapia antirretroviral e da infecção pelo HPV sobre as células dendríticas epiteliais e subepiteliais. MÉTODOS: Um estudo prospectivo foi realizado para analisar morfometricamente o volume relativo das células dendríticas e as relações entre neoplasia intraepitelial anal e o câncer em pacientes HIV positivos da Fundação de Medicina Tropical do Amazonas, Brasil.Todos os pacientes foram submetidos a biópsia da mucosa retal para realizar uma análise clássica histopatológica e imunohistoquímica utilizando anticorpos contra anti-CD1a e anti-DC-SIGN, para a quantificação morfométrica das células dendríticas. RESULTADOS: Os pacientes HIV negativos apresentaram densidade das DC CD1a+ significativamente maior do que a dos pacientes HIV positivos (3,75 versus 2,54) (p:0,018), e os pacientes com severa apresentaram correlação das DC CD1a com os níveis de células TCD4(p:0,04) assim como a carga viral do HIV-1 (p:0,035). Observamos no subgrupo HIV-positivo/HAART positivo elevação não significativa na mediana da densidade das DC CD1a+ em relação ao grupo HIV-positivo/HAART negativo. As DC CD1a+ também se elevaram nos pacientes HIV negativo portadores de condiloma anorretal(2,33 para 3,53; p:0,05), com efeito inverso nos pacientes HIV positivos. CONCLUSÕES: Nossos dados confirmam a potencialização da ação sinérgica representada pela coinfecção HIV-HPV sobre o epitélio anal, fragilizando as DC em sua função primordial de vigilância imune. Notoriamente nos pacientes com neoplasia intraepithelial anal grave, a densidade das DC CD1a+ epiteliais sofreu influência da carga viral do HIV-1.(AU)
Assuntos
Humanos , HIV , Mucosa/anormalidades , Canal Anal/anatomia & histologia , NeoplasiasRESUMO
PURPOSE: To morphometrically quantify CD1a+ dentritic cells and DC-SIGN+ dendritic cells in HIV-positive patients with anal squamous intraepithelial neoplasia and to evaluate the effects of HIV infection, antiretroviral therapy and HPV infection on epithelial and subepithelial dendritic cells. METHODS: A prospective study was performed to morphometrically analyze the relative volume of the dendritic cells and the relationship between anal intraepithelial neoplasia and cancer in HIV-positive patients from the Tropical Medicine Foundation of Amazonas, Brazil. All patients were submitted to biopsies of anorectal mucosa to perform a classic histopathological and immunohistochemical analysis, employing antibodies against CD1a and DC-SIGN for the morphometric quantification of dendritic cells. RESULTS: HIV-negative patients displayed a CD1a DC density significantly higher than that of HIV-positives patients (3.75 versus 2.54) (p=0.018), and in patients with severe anal intraepithelial neoplasia had correlated between DC CD1a density with levels of CD4 + cells (p: 0.04) as well as the viral load of HIV-1 (p: 0.035). A not significant rise in the median density of CD1a+ DC was observed in the HIV positive/ HAART positive subgroup compared to the HIV positive/ HAART negative subgroup. The CD1a+ DC were also significantly increased in HIV-negative patients with anorectal condyloma (2.33 to 3.53; p=0.05), with an opposite effect in HIV-positive patients. CONCLUSIONS: Our data support an enhancement of the synergistic action caused by HIV-HPV co-infection on the anal epithelium, weakening the DC for its major role in immune surveillance. Notoriously in patients with severe anal intraepithelial neoplasia, the density of CD1a+ epithelial dendritic cells was influenced by the viral load of HIV-1. Our study describes for the first time the density of subepithelial DC-SIGN+ dendritic cells in patients with anal severe anal intraepithelial neoplasia and points to the possibility that a specific therapy for HIV induces the recovery of the density of epithelial DC.
OBJETIVO: Quantificar morfometricamente as células dendríticas DC CD1a+ e DC DC-SIGN+ em pacientes HIV positivos portadores de neoplasia escamosa intraepitelial anal e avaliar os efeitos da infecção pelo HIV, da terapia antirretroviral e da infecção pelo HPV sobre as células dendríticas epiteliais e subepiteliais. MÉTODOS: Um estudo prospectivo foi realizado para analisar morfometricamente o volume relativo das células dendríticas e as relações entre neoplasia intraepitelial anal e o câncer em pacientes HIV positivos da Fundação de Medicina Tropical do Amazonas, Brasil.Todos os pacientes foram submetidos a biópsia da mucosa retal para realizar uma análise clássica histopatológica e imunohistoquímica utilizando anticorpos contra anti-CD1a e anti-DC-SIGN, para a quantificação morfométrica das células dendríticas. RESULTADOS: Os pacientes HIV negativos apresentaram densidade das DC CD1a+ significativamente maior do que a dos pacientes HIV positivos (3,75 versus 2,54) (p:0,018), e os pacientes com severa apresentaram correlação das DC CD1a com os níveis de células TCD4(p:0,04) assim como a carga viral do HIV-1 (p:0,035). Observamos no subgrupo HIV-positivo/HAART positivo elevação não significativa na mediana da densidade das DC CD1a+ em relação ao grupo HIV-positivo/HAART negativo. As DC CD1a+ também se elevaram nos pacientes HIV negativo portadores de condiloma anorretal(2,33 para 3,53; p:0,05), com efeito inverso nos pacientes HIV positivos. CONCLUSÕES: Nossos dados confirmam a potencialização da ação sinérgica representada pela coinfecção HIV-HPV sobre o epitélio anal, fragilizando as DC em sua função primordial de vigilância imune. Notoriamente nos pacientes com neoplasia intraepithelial anal grave, a densidade das DC CD1a+ epiteliais sofreu influência da carga viral do HIV-1. Nosso estudo descreveu pela primeira vez a densidade das DC subepiteliais DC-SIGN+ em pacientes com neoplasia intraepithelial anal severa e apontamos para a possibilidade de que a terapia específica para o HIV induza a recuperação da densidade das DC epiteliais.
Assuntos
Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Neoplasias do Ânus/patologia , Carcinoma in Situ/patologia , Carcinoma de Células Escamosas/patologia , Condiloma Acuminado/patologia , Células Dendríticas/patologia , Soropositividade para HIV/patologia , Terapia Antirretroviral de Alta Atividade , Canal Anal/patologia , Canal Anal/virologia , Neoplasias do Ânus/imunologia , Neoplasias do Ânus/virologia , Estudos de Casos e Controles , Carcinoma in Situ/imunologia , Carcinoma in Situ/virologia , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/virologia , Condiloma Acuminado/imunologia , Condiloma Acuminado/virologia , Células Dendríticas/imunologia , Células Dendríticas/virologia , Soropositividade para HIV/tratamento farmacológico , Soropositividade para HIV/imunologia , Imuno-Histoquímica , Imunidade Celular/imunologia , Mucosa/imunologia , Estudos Prospectivos , Infecções por Papillomavirus/imunologia , Infecções por Papillomavirus/patologiaRESUMO
PURPOSE: To morphometrically quantify CD1a+ dentritic cells and DC-SIGN+ dendritic cells in HIV-positive patients with anal squamous intraepithelial neoplasia and to evaluate the effects of HIV infection, antiretroviral therapy and HPV infection on epithelial and subepithelial dendritic cells. METHODS: A prospective study was performed to morphometrically analyze the relative volume of the dendritic cells and the relationship between anal intraepithelial neoplasia and cancer in HIV-positive patients from the Tropical Medicine Foundation of Amazonas, Brazil. All patients were submitted to biopsies of anorectal mucosa to perform a classic histopathological and immunohistochemical analysis, employing antibodies against CD1a and DC-SIGN for the morphometric quantification of dendritic cells. RESULTS: HIV-negative patients displayed a CD1a DC density significantly higher than that of HIV-positives patients (3.75 versus 2.54) (p=0.018), and in patients with severe anal intraepithelial neoplasia had correlated between DC CD1a density with levels of CD4 + cells (p: 0.04) as well as the viral load of HIV-1 (p: 0.035). A not significant rise in the median density of CD1a+ DC was observed in the HIV positive/ HAART positive subgroup compared to the HIV positive/ HAART negative subgroup. The CD1a+ DC were also significantly increased in HIV-negative patients with anorectal condyloma (2.33 to 3.53; p=0.05), with an opposite effect in HIV-positive patients. CONCLUSIONS: Our data support an enhancement of the synergistic action caused by HIV-HPV co-infection on the anal epithelium, weakening the DC for its major role in immune surveillance. Notoriously in patients with severe anal intraepithelial neoplasia, the density of CD1a+ epithelial dendritic cells was influenced by the viral load of HIV-1. Our study describes for the first time the density of subepithelial DC-SIGN+ dendritic cells in patients with anal severe anal intraepithelial neoplasia and points to the possibility that a specific therapy for HIV induces the recovery of the density of epithelial DC.
Assuntos
Neoplasias do Ânus/patologia , Carcinoma in Situ/patologia , Carcinoma de Células Escamosas/patologia , Condiloma Acuminado/patologia , Células Dendríticas/patologia , Soropositividade para HIV/patologia , Adolescente , Adulto , Idoso , Canal Anal/patologia , Canal Anal/virologia , Terapia Antirretroviral de Alta Atividade , Neoplasias do Ânus/imunologia , Neoplasias do Ânus/virologia , Carcinoma in Situ/imunologia , Carcinoma in Situ/virologia , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/virologia , Estudos de Casos e Controles , Condiloma Acuminado/imunologia , Condiloma Acuminado/virologia , Células Dendríticas/imunologia , Células Dendríticas/virologia , Feminino , Soropositividade para HIV/tratamento farmacológico , Soropositividade para HIV/imunologia , Humanos , Imunidade Celular/imunologia , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Mucosa/imunologia , Infecções por Papillomavirus/imunologia , Infecções por Papillomavirus/patologia , Estudos Prospectivos , Adulto JovemRESUMO
The gill morphologies of six species of potamotrygonid freshwater stingrays from the Amazon basin were investigated using light and electron microscopy. Some unique features were found in the potamotrygonid gill: (1) fingerlike protuberances on the gill filament, (2) an Alcian blue/periodic acid-Schiff-positive histochemical reaction for several cell layers in the gill epithelium (except the basal ones), (3) pavement cells with numerous subapical mucous vesicles, (4) very large mucous cells, and (5) follicular Na(+)/K(+)-ATPase-rich (NKA-rich) mitochondria-rich cells (MRCs) in Potamotrygon sp. (known as the cururu ray). The fingerlike protuberances may constitute an additional resistance to water flow, helping to drive water through the lamellae. The secretion of a mucous substance by the pavement cells and mucous cells may help to protect the gills against mechanical injury and pathogens and aid in osmoregulation in the dilute water of the Amazon basin. All MRCs possess enfolded basolateral membranes and have poorly developed or absent tubular systems. NKA-rich MRCs are located high in the basolateral membrane. The cururu ray, which is endemic to the Rio Negro, has follicular NKA-rich MRCs (8-12 cells in cross section) that share the same apical pit in the filament; this may be considered to be an autapomorphy. The combination of these branchial characteristics may have favored tolerance to the freshwater environment during the evolution and diversification of potamotrygonids throughout the Amazon basin.
Assuntos
Adaptação Fisiológica/fisiologia , Elasmobrânquios/anatomia & histologia , Brânquias/anatomia & histologia , Animais , Brasil , Elasmobrânquios/metabolismo , Elasmobrânquios/fisiologia , Células Epiteliais/enzimologia , Células Epiteliais/fisiologia , Brânquias/citologia , Brânquias/enzimologia , Brânquias/fisiologia , Imuno-Histoquímica , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Mitocôndrias/enzimologia , Mitocôndrias/fisiologia , Rios , ATPase Trocadora de Sódio-Potássio/fisiologia , Equilíbrio Hidroeletrolítico/fisiologiaRESUMO
Previous morphometric methods for estimation of the volume of components, surface area and thickness of the diffusion barrier in fish gills have taken advantage of the highly ordered structure of these organs for sampling and surface area estimations, whereas the thickness of the diffusion barrier has been measured orthogonally on perpendicularly sectioned material at subjectively selected sites. Although intuitively logical, these procedures do not have a demonstrated mathematical basis, do not involve random sampling and measurement techniques, and are not applicable to the gills of all fish. The present stereological methods apply the principles of surface area estimation in vertical uniform random sections to the gills of the Brazilian teleost Arapaima gigas. The tissue was taken from the entire gill apparatus of the right-hand or left-hand side (selected at random) of the fish by systematic random sampling and embedded in glycol methacrylate for light microscopy. Arches from the other side were embedded in Epoxy resin. Reference volume was estimated by the Cavalieri method in the same vertical sections that were used for surface density and volume density measurements. The harmonic mean barrier thickness of the water-blood diffusion barrier was calculated from measurements taken along randomly selected orientation lines that were sine-weighted relative to the vertical axis. The values thus obtained for the anatomical diffusion factor (surface area divided by barrier thickness) compare favourably with those obtained for other sluggish fish using existing methods.