RESUMO
The aim of the present study was to compare healing obtained with biomembranes with the natural healing process (sham) using biochemical and immunohistological assays. C57BL/6 mice were divided into 4 groups of 15 mice each and received different subcutaneous implants: natural latex biomembrane (NLB), denatured latex (DL), expanded polytetrafluorethylene (ePTFE), or sham. On the 2nd, 7th, and 14th days post-treatment, 5 mice per group were sacrificed and biopsied for the following measurements: oxidative stress based on malondialdehyde (MDA), myeloperoxidase (MPO) and hydrogen peroxide by the method of ferrous oxidation-xylenol orange (FOX), as well as glutathione and total proteins; histological evaluation to enumerate inflammatory cells, fibroblasts, blood vessels, and collagen, and immunohistochemical staining for inducible nitric oxide synthase, interleukin-1β, vascular endothelial growth factor (VEGF), and transforming growth factor-β1 (TGF-β1). On day 2 post-treatment, NLB stimulated a dense inflammatory infiltrate mainly consisting of polymorphonuclear cells, as indicated by increased MPO (P < 0.05), but oxidative stress due to MDA was not observed until the 7th day (P < 0.05). The number of blood vessels was greater in NLB (P < 0.05) and DL (P < 0.05) mice compared to sham animals on day 14. NLB induced fibroplasia by day 14 (P < 0.05) with low expression of TGF-β1 and collagenesis. Thus, NLB significantly induced the inflammatory phase of healing mediated by oxidative stress, which appeared to influence the subsequent phases such as angiogenesis (with low expression of VEGF) and fibroplasia (independent of TGF-β1) without influencing collagenesis.
Assuntos
Animais , Masculino , Camundongos , Materiais Biocompatíveis/uso terapêutico , Látex/uso terapêutico , Membranas Artificiais , Estresse Oxidativo/fisiologia , Politetrafluoretileno/uso terapêutico , Cicatrização/fisiologia , Imuno-Histoquímica , Inflamação/fisiopatologia , Estresse Oxidativo/efeitos dos fármacos , Cicatrização/efeitos dos fármacosRESUMO
The aim of the present study was to compare healing obtained with biomembranes with the natural healing process (sham) using biochemical and immunohistological assays. C57BL/6 mice were divided into 4 groups of 15 mice each and received different subcutaneous implants: natural latex biomembrane (NLB), denatured latex (DL), expanded polytetrafluorethylene (ePTFE), or sham. On the 2nd, 7th, and 14th days post-treatment, 5 mice per group were sacrificed and biopsied for the following measurements: oxidative stress based on malondialdehyde (MDA), myeloperoxidase (MPO) and hydrogen peroxide by the method of ferrous oxidation-xylenol orange (FOX), as well as glutathione and total proteins; histological evaluation to enumerate inflammatory cells, fibroblasts, blood vessels, and collagen, and immunohistochemical staining for inducible nitric oxide synthase, interleukin-1ß, vascular endothelial growth factor (VEGF), and transforming growth factor-ß1 (TGF-ß1). On day 2 post-treatment, NLB stimulated a dense inflammatory infiltrate mainly consisting of polymorphonuclear cells, as indicated by increased MPO (P < 0.05), but oxidative stress due to MDA was not observed until the 7th day (P < 0.05). The number of blood vessels was greater in NLB (P < 0.05) and DL (P < 0.05) mice compared to sham animals on day 14. NLB induced fibroplasia by day 14 (P < 0.05) with low expression of TGF-ß1 and collagenesis. Thus, NLB significantly induced the inflammatory phase of healing mediated by oxidative stress, which appeared to influence the subsequent phases such as angiogenesis (with low expression of VEGF) and fibroplasia (independent of TGF-ß1) without influencing collagenesis.
Assuntos
Materiais Biocompatíveis/uso terapêutico , Látex/uso terapêutico , Membranas Artificiais , Estresse Oxidativo/fisiologia , Politetrafluoretileno/uso terapêutico , Cicatrização/fisiologia , Animais , Imuno-Histoquímica , Inflamação/fisiopatologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Estresse Oxidativo/efeitos dos fármacos , Cicatrização/efeitos dos fármacosRESUMO
OBJECTIVE: To verify the validity of measuring the levels of Mycobacterium leprae-specific anti-phenolic glycolipid (PGL)-I antibody, neopterin, a product of activated macrophages, and C-reactive protein (CRP), an acute phase protein, in serial serum samples from patients for monitoring the leprosy spectrum and reactions during the course of multi-drug treatment (MDT). METHODS: Twenty-five untreated leprosy patients, 15 multi-bacillary (MB) and 10 paucibacillary (PB), participated. Eight patients developed reversal reaction and five developed erythema nodosum leprosum (ENL) during follow-up. The bacterial index (BI) in slit-skin smears was determined at diagnosis and blood samples collected by venipuncture at diagnosis and after 2, 4, 6 and 12 months of MDT. PGL-I antibody and neopterin were measured by enzyme-linked immunosorbent assay, whereas the CRP levels were measured by the latex agglutination method. RESULTS: The levels of PGL-I antibodies and neopterin were higher in the sera of MB than PB patients, which correlated with the patients' BI. The serum levels of CRP did not differ significantly between the MB and PB patients. The serum levels of PGL-I and neopterin were no higher in reactional patients than non-reactional patients prone to such reactions. However, ENL patients had higher serum CRP levels than non-reactional MB patients. The serum PGL-I antibody levels declined significantly during MDT, in contrast to neopterin and CRP levels. CONCLUSION: Measuring the serum levels of PGL-I antibodies and neopterin appeared to be useful in distinguishing MB from PB patients, whereas monitoring the levels of PGL-I antibodies appeared to be useful in monitoring MB patients on MDT. Measuring serum CRP, although not useful in monitoring the patients, has limited significance in detecting ENL reactional patients.
Assuntos
Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Proteína C-Reativa/metabolismo , Glicolipídeos/imunologia , Hanseníase Dimorfa/sangue , Hanseníase Tuberculoide/sangue , Neopterina/sangue , Adulto , Idoso , Feminino , Humanos , Hansenostáticos/efeitos adversos , Hansenostáticos/uso terapêutico , Hanseníase Dimorfa/tratamento farmacológico , Hanseníase Dimorfa/imunologia , Hanseníase Tuberculoide/tratamento farmacológico , Hanseníase Tuberculoide/imunologia , Masculino , Pessoa de Meia-IdadeRESUMO
Food utilization by the larvae of Toxorhynchites splendens (Wiedemann) was studied in the laboratory by offering larvae of Aedes aegypti Linnaeus, Anopheles stephensi (Liston), and Culex quinquefasciatus (Say). Quantitative analyses of data indicated that immature development was significantly faster with increase in food availability. The regression analysis showed that the degrees of the relationship between immature duration (Id) and food availability were higher when offered early instars of prey (first and second instars) than late instars. Consumption rate (Cr) of the predator increased with increase in food availability and this relationship was highly significant when larvae of An. stephensi were offered as food. Consumption rate to food level decreased with increase in the age class of the prey. There was a significant negative correlation between Id and Cr. This aspect helps to increase population turnover of T. splendens in a shorter period when the prey is abundant. Conversely, the predator compensated the loss in daily food intake at low food level by extending Id thereby attains the minimum threshold pupal weight for adult emergence. There was an increase in the relative growth rate (RGR) of the predator when An. stephensi was offered as prey and this was related to the high protein content of the prey per body weight. There was a positive correlation between Cr and RGR. This adaptive life characteristic strategy of this predator is useful for mass-rearing for large scale field release programmes in the control of container breeding mosquitoes is discussed.
Assuntos
Culicidae/crescimento & desenvolvimento , Comportamento Predatório/fisiologia , Animais , Regulação do Apetite/fisiologia , Culicidae/fisiologia , Comportamento Alimentar/fisiologia , Larva , Valor NutritivoRESUMO
In view of the recent discovery of rickettsial endosymbionts, Wolbachia in lymphatic filarial parasites, Wuchereria bancrofti and Brugia malayi and subsequently of their vital role in the survival and development of the latter, antibiotics such as tetracycline are being suggested for the treatment of lymphatic filariasis, by way of eliminating the endosymbiont. But, it is essential to assess their presence in parasites from areas endemic for lymphatic filariasis before such a new control tool is employed. In the present communication, we report the detection of Wolbachia endosymbionts in microfilariae of W. bancrofti parasites collected from geographically distant locations of India, such as Pondicherry (Union Territory), Calicut (Kerala), Jagadalpur (Madhya Pradesh), Thirukoilur (TamilNadu), Chinnanergunam (TamilNadu), Rajahmundry (Andhra Pradesh), and Varanasi (Uttar Pradesh), using Wolbachia specific 16S rDNA polymerase chain reaction.
Assuntos
Animais , Microfilárias , Wolbachia , Wuchereria bancrofti , DNA Bacteriano , DNA Ribossômico , Índia , Reação em Cadeia da PolimeraseRESUMO
In view of the recent discovery of rickettsial endosymbionts, Wolbachia in lymphatic filarial parasites, Wuchereria bancrofti and Brugia malayi and subsequently of their vital role in the survival and development of the latter, antibiotics such as tetracycline are being suggested for the treatment of lymphatic filariasis, by way of eliminating the endosymbiont. But, it is essential to assess their presence in parasites from areas endemic for lymphatic filariasis before such a new control tool is employed. In the present communication, we report the detection of Wolbachia endosymbionts in microfilariae of W. bancrofti parasites collected from geographically distant locations of India, such as Pondicherry (Union Territory), Calicut (Kerala), Jagadalpur (Madhya Pradesh), Thirukoilur (TamilNadu), Chinnanergunam (TamilNadu), Rajahmundry (Andhra Pradesh), and Varanasi (Uttar Pradesh), using Wolbachia specific 16S rDNA polymerase chain reaction.
Assuntos
Microfilárias/microbiologia , Wolbachia/isolamento & purificação , Wuchereria bancrofti/microbiologia , Animais , DNA Bacteriano/análise , DNA Ribossômico/análise , Índia , Reação em Cadeia da PolimeraseRESUMO
The LYMFASIM modeling framework for the transmission and control of the tropical parasitic disease lymphatic filariasis is described and its use in the context of an endemic community in north-eastern Brazil is illustrated. Lymphatic filariasis is a disease with a complex natural history with many unknowns. This complicates decision making with respect to control strategies. With LYMFASIM, a variety of hypotheses can be tested about the life history of the parasite Wuchereria bancrofti, its transmission from man to man through mosquitoes, the role of the immune system in regulating parasite numbers, the development of disease symptoms, and the effects of control measures (drug treatment or mosquito control). The implications of alternative assumptions and uncertainty about the quantification of parameters for the effectiveness of control strategies can be investigated. Thanks to the use of stochastic microsimulation, LYMFASIM is highly flexible and can be adapted and extended as new knowledge emerges.
Assuntos
Simulação por Computador , Tomada de Decisões Assistida por Computador , Filariose Linfática/prevenção & controle , Modelos Biológicos , Adolescente , Animais , Brasil , Criança , Pré-Escolar , Filariose Linfática/epidemiologia , Filariose Linfática/parasitologia , Filariose Linfática/fisiopatologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Wuchereria bancrofti/fisiologiaAssuntos
Anticorpos Monoclonais/imunologia , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/química , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/química , Dermatopatias/imunologia , Granuloma/imunologia , Mycobacterium leprae/imunologia , Peso Molecular , Reações CruzadasRESUMO
Mycobacterium bovis-BCG was sonified and centrifuged at 90,000g for 2h to obtain pellet (P90) and supernatant (S90), and bacilli broken by chilled X-press were fractionated into cell wall (CW), plasma membrane and cytosol. Rabbits were immunized with P90, S90 and whole sonified-broken BCG. The antigenic make-up of these antisera. These analysed by cross-immunoelectrophoresis using the antisera. These subcellular preparations were also used for detecting circulating antibodies in the sera of 45 leprosy patients by Ouchterlony´s technique and immunoelectrophoresis. Although there were many antigenic determinants common to more than one fraction, some components were found in only one fraction. Furthermore, different leprosy patients showed different patterns of antibody response to the antigens in the different fractions. In addition, these fractions were also used to assess cell-mediated immunity in mice sensitized with lyophilized whole BCG as well as with irradiated Mycobacterium. it was found that BCG, intraperitoneally injected, engedered different specifically antigen-sensitized populations of lymphocytes in the spleen and lymph node. Moreover, a certain degree of antigenic cross-reactivity was observed between BCG and M. leprae, possibly at the T-cell level. In vitro experiments suggested that the CW stimulated B cells, indicating a mitogenic ctivity leading to polyclonal activation. Finally, in vitro priming experiments established that these subcellular fractions could sensitize human peripheral lymphocytes, and upon secondary culture all primed cells were able to respond to homologous preparations but not always to heterologous preparations, thus offering a means to distinguish antigens of interest from those antigenically less complicated fractions at the T-cell level.