Histopathology and immunocytochemical study of type 3 and type 4 complement receptors in the liver and spleen of dogs naturally and experimentally infected with Leishmania (Leishmania) chagasi

The objective of this study was to compare the histopathological changes and expression of CR3 and CR4 in the liver and spleen of dogs naturally and experimentally infected with L. chagasi. The basic histopathological lesions observed mainly in naturally infected dogs were: epithelioid hepatic granulomas, hyperplasia and hypertrophy of Kupffer cells, Malpigui follicles and mononucleated cells of the red pulp of the spleen. Sections from the liver and spleen by immunocytochemistry technique showed the presence of CD11b,c\CD 18 antigens in the control and infected animals and no qualitative or quantitative differences in the liver. Nevertheless, CD18 was always increased in the spleen of naturally and experimentally infected dogs. These results indicate that there is a difference in the activaton of CD 18 in both experimental and natural cases of canine visceral leishmaniasis that should play an important role in the immunological response to Leishmania chagasi infection.

Effect of b-Propiolactone treatment on the complement activation mediated by equine antisera

Reduction of complement activation through an alteration of the Fc fragment of immunoglobulins by b-propiolactone treatment was carried out in equine antisera raised against rabies virus, Bothrops venoms and diphtherial toxin. Results were evaluated by means of an anaphylactic test performed on guinea-pigs, and compared to the ones obtained with the same sera purified by saline precipitation (ammonium sulfate), followed or not by enzymatic digestion with pepsin. Protein purity levels for antibothropic serum were 184.5 mg/g and 488.5 mg/g in b-propiolactone treated and pepsin-digested sera, respectively. The recovery of specific activity was 100% and 62.5% when using antibothropic serum treated by b-propiolactone and pepsin digestion, respectively. The antidiphtherial and anti-rabies sera treated with b-propiolactone and pepsin presented protein purity levels of 5,698 and 7,179 Lf/g, 16,233 and 6,784 IU/g, respectively. The recovery of specific activity for these antisera were 88.8%, 77.7%, 100% and 36,5%, respectively. b-propiolactone treatment induced a reduction in complement activation, tested "in vivo", without significant loss of biological activity. This treatment can be used in the preparation of heterologous immunoglobulins for human use.

In vitro and in situ activation of the complement system by the fungus Lacazia loboi

Since there are no studies evaluating the participation of the complement system (CS) in Jorge Lobo's disease and its activity on the fungus Lacazia loboi, we carried out the present investigation. Fungal cells with a viability index of 48% were obtained from the footpads of BALB/c mice and incubated with a pool of inactivated serum from patients with the mycosis or with sterile saline for 30 min at 37 ºC. Next, the tubes were incubated for 2 h with a pool of noninactivated AB+ serum, inactivated serum, serum diluted in EGTA-MgCl2, and serum diluted in EDTA. The viability of L. loboi was evaluated and the fungal suspension was cytocentrifuged. The slides were submitted to immunofluorescence staining using human anti-C3 antibody. The results revealed that 98% of the fungi activated the CS by the alternative pathway and no significant difference in L. loboi viability was observed after CS activation. In parallel, frozen histological sections from 11 patients were analyzed regarding the presence of C3 and IgG by immunofluorescence staining. C3 and IgG deposits were observed in the fungal wall of 100% and 91% of the lesions evaluated, respectively. The results suggest that the CS and immunoglobulins may contribute to the defense mechanisms of the host against L. loboi.

Longitudinal study of the indirect immunofluorescence and complement fixation tests for diagnosis of chagas' disease in immunosuppressed patients submitted to renal transplantation

Clinical and serological follow-up of 7 patients submitted to renal transplantation and presenting positive serological reactions to Chagas 'disease before immunossupression did not show significant changes in indirect immunofluorescence and complement fixation titres for Chagas ' disease, or signs and symptoms indicating exacerbation of the disease during follow- up. In addition, 18 of 66 recipients of renal transplants considered to be non-chagasic before immunosuppression showed at least one positive result to the indirect immunofluorescence test for Chagas ' disease during the study period. The results suggest that the immunosuppression State induced in chagasic patients submitted to renal transplant did notpromoted exacerbation of the chronic infection in these patients and not interfere with the serological response of chronic chagasics, thus permitting the use of these serologic reactions for diagnostic purposes in these cases. However, the positive results ofthe indirect immunofluorescence test in non- chagasic patients indicate the needforjudicious interpretation ofthe indirect immunofluorescence test for the diagnosis of Chagas' disease in renal transplanted patients.

Discriminação de fitofisionomias de floresta de várzea a partir do algoritmo Iterated Conditional Modes aplicado aos dados SAR/R99 (QUAD-POL/Banda L)

Utilizando-se dados do sensor aerotransportado SAR R99, adquiridos na banda L (1,28 GHz) em amplitude e com quatro polarizações (HH, VV, HV e VH), avaliou-se a distinção de fitofisionomias de floresta de várzea existentes nas Reservas de Desenvolvimento Sustentável Amanã e Mamirauá e áreas adjacentes, com a aplicação do algoritmo Iterated Conditional Modes (ICM) de classificação polarimétrica pontual/contextual. Os resultados mostraram que o uso das distribuições multivariadas em amplitude, conjuntamente com uma banda de textura, produziu classificações de qualidade superior àquelas obtidas com dados polarimétricos uni/bivariados. Esta abordagem permitiu a obtenção de um índice Kappa de 0,8963, discriminando as três classes vegetacionais de interesse, comprovando assim o potencial dos dados do SAR R99 e do algoritmo ICM no mapeamento de florestas de várzea da Amazônia.

Adaptação e aplicabilidade de um algoritmo de diurético para pacientes com insuficiência cardíaca

FUNDAMENTO: Estados congestivos podem ser identificados e manejados através de algoritmos como o Diuretic Treatment Algorithm (DTA) para ajuste de diurético por telefone, com enfoque na avaliação clínica. Porém, o DTA está disponível somente em língua inglesa. OBJETIVO: Adaptar o DTA e testar sua aplicabilidade para uso no Brasil em pacientes ambulatoriais com IC. MÉTODOS: Seguiram-se as etapas de tradução, síntese, retrotradução, avaliação por comitê de especialistas e pré-teste (aplicabilidade clínica por meio de ensaio clínico randomizado). O DTA foi denominado, na versão para o Brasil, algoritmo de ajuste de diurético (AAD). Os pacientes foram randomizados para grupo intervenção (GI) - ajuste de diurético conforme o AAD - ou grupo controle (GC) - ajuste convencional. Foi avaliado o escore clínico de congestão (ECC) e o peso para ambos os grupos. RESULTADOS: Foram realizadas 12 modificações no DTA. Incluíram-se 34 pacientes. Para aqueles congestos, o aumento de diurético guiado pelo AAD resultou em maior resolução da congestão, com redução de dois pontos no ECC para 50% da amostra -2 (-3,5; -1,0), enquanto a mediana para o GC foi 0 (-1,25; -1,0), (p < 0,001). A mediana de variação de peso foi maior no GI -1,4 (-1,7; -0,5) quando comparado ao GC 0,1 (1,2; -0,6), p = 0,001. CONCLUSÕES: O ADD mostrou-se aplicável na prática clínica após adaptação e parece resultar em melhor controle da congestão em pacientes com IC. A efetividade clínica da ferramenta merece ser testada em amostra maior de pacientes visando sua validação para uso no Brasil (Universal Trial Number: U1111-1130-5749) (Arq Bras Cardiol. 2013; [online]. ahead print, PP.0-0).

Contribution to the study of immune hemolysis by toad complement

EA (sheep erythrocytes carrying rabbit antibody) are lysed by toad complement under optimal conditions which include a low concentration of cells (1.54 x 10*8/ml), a low temperature of incubation (30°C) and the same amounts of Ca++ and Mg++ as required for the titration of guinea-pig complement. Kinetic studies of the role of cations mentioned above in immune lysis by toad C have disclosed a fundamental difference as compared to guinea-pig C. In a limited complement system, the lysis by amphibian C is completely blocked by EDTA, even when the chelating agent is added as late as 15 minutes after zero-time. Inhibition by EGTA is only partial and the findings suggest that Mg++ is required not only at the beginning, but also at late stages of the lytic process. It has been speculated that the activation of amphibian complement proceeds mainly by the alternative pathway.

Decay accelerating factor (DAF) as the host antigen with protective activity to complement killing of schistosomula

The acquisition of host antigens by Schistosoma mansoni was studied by evaluating the resistance of schistosomula to the complement attack mediated by lethal antibody. Schistosomula cultured for 24 hours with intact human erythrocytes (N-HuE) or ghosts of any type of ABO or Rh blood group, showed a marked resistance to complement damage. Sheep red blood cells, pronase-treated N-HuE or erythrocytes from patients with paroxysmal nocturnal hemoglobinuria, which are complement-sensitive cells, were unable to protect schistosomula. Schistosomula protected by N-HuE became again susceptible to complement killing after incubation with a monoclonal antibody anti-DAF. These results indicate that, in vitro, host DAF from N-HuE can be acquired by schistosomula surface in a biological active form that protects the parasite from the complement lesion.

Mechanisms of evasion of Schistosoma mansoni schistosomula to the lethal activity of complement

Schistosomula of Schistosoma mansoni became resistant to antibody-dependent complement damage in vitro after pre-incubation with normal human erythrocytes (NHuE) whatever the ABO or Rh blood group. Resistant parasites were shown to acquire host decay accelerating factor (DAF) , a 70 kDa glycoprotein attached to the membrane of NHue by a GPI anchor. IgG2a mAb anti-human DAF (IA10) immunoprecipitated a 70 kDa molecule from 125I-labeled schistosomula pre-incubated with NHuE and inhibited their resistance to complement-dependent killing in vtro. Incubationof schistosomula with erytrocytes from patients with paroxsimal nocturnal hemoglobinuria (PNHE) or SRBC, wich are DAF-deficient, did not protect the parasites from complement lesion. Supernatant of 100,000 x g collected from NHuE incubated for 24 h in defined medium was shown to contain a soluble form of DAF and to protect schistosomula from complement killing. Schistosomula treated with trypsin before incubation with NHuE ghosts did not become resistant to complement damage. On the other hand, pre-treatment with chymotrypsin did not interfere with the acquisition of resistance by the schistosomula. These results indicate that, in vitro, NHuE DAF can be transferred to schistosomula in a soluble form and that the binding of this molecule to the parasite surface is dependent upon trypsin-sensitive chymotrypsin-insensitive polipeptide(s) present on the surface of the worm.

Complete immunization against Trypanosoma cruzi verified in individual mice by complement-mediated lysis

Experimental systems to assay immunity against Trypanosoma cruzi usually demonstrate partial resistance without excluding the establishment of sub-patent infections in protected animals. To test whether Swiss mice immunized with attenuated parasites might develop complete resistance against virulent T. cruzi, experiments were performed involving challenge with low numbers of parasites, enhancement of local inflammation and the combination of natural and acquired resistance. Absence of infection was established after repeated negative parasitological tests (including xenodiagnosis and hemoculture), and lack of lytic antibody was tested by complement mediated lysis. Immunization with 10(7) attenuated epimastigotes conferred protection against the development of high levels of parasitemia after challenge with Tulahuen strain, but was unable to reduce the number of infected animals. However, when a strong, delayed-type hypersensitivity reaction was triggered at the site of infection by injecting a mixture of virulent and attenuated T. cruzi, a significant proportion of immunized animals remained totally free of virulent infection. The same result was obtained when the immunization experiment was performed in four month old Swiss mice, displaying a relatively high natural resistance and challenged with wild, vector-borne parasites. These experiments demonstrate that complete resistance against T. cruzi can be obtained in a significant proportion of animals, under conditions which replicate natural, vector delivered infection by the parasite.

Levels of complement C3 and C4 components in Amerindians living in an area with high prevalence of tuberculosis

The levels of complement C3 and C4 components were determined in non-indigenous (creoles) and indigenous (Warao) populations, the latter with an extremely high tuberculosis (TB) rate. Serum samples from 209 adults were studied and classified in 4 groups taking into account tuberculin skin tests (TST): (1) the group of Warao patients (58 positive for the TST, WP TST+ and 9 negative for the TST, WP TST-), (2) the group of creole patients (34 positive for the TST, CP TST+ and 9 negative for the TST, CP TST-), (3) the group of healthy Warao controls (38 positive and 14 negative for TST, WC TST+ and WC TST-, respectively), (4) the creole controls (26 positive and 21 negative for the TST, CC TST+ and CC TST-, respectively). With respect to the results concerning the measurement of both complement C3 and C4 components with the exception of the WC TST and the CC groups, the WP TST+ and WP TST- as well as WC TST+ groups showed a significant frequency of individuals with decreased levels of complement C3 component (20.6, 33.3, and 26.3%, respectively) and also C4 component (12.0, 11.1, and 13.3%, respectively) in comparison to both creole patients (CP TST+, 8.82% and CP TST-, 0% and CP TST+, 5.88% and CP TST-, 0%) for C3 and C4, respectively. The study of these parameters carried out in 15 Warao subjects with active infection, before and after anti-TB chemotherapy,statisticallyconfirmedthat the effective chemotherapy did not restore normal levels of the complement C3 and C4 components among Warao patients. Aditional tests for hepatitis B or hepatitis C infection, and the profile of the hepatic proteins were not associated to the deficiency in production of the complement components.In conclusion, the results show that within the Warao population, a high percentage of subjects exhibit decreased levels of both complement C3 and C4 components independent of latent or active infection and the status of TST.

Um algoritmo prático para determinação da variância do estimador de um contraste de parâmetros

Testes de significância de contrastes de efeitos de níveis de um fator ou de combinações de níveis de dois ou mais fatores requerem o conhecimento das estimativas das variâncias dos estimadores desses contrastes. As expressões dessas estimativas para experimentos com delineamentos complexos, em geral, não são disponíveis em textos. Sua derivação algébrica a partir da equação do modelo estatístico é trabalhosa. Expõe-se um algoritmo prático para a obtenção da variância do estimador de qualquer contraste de parâmetros para delineamentos completos balanceados. O algoritmo baseia-se nos valores esperados dos quadrados médios.

Séries temporais de NDVI do sensor SPOT Vegetation e algoritmo SAM aplicados ao mapeamento de cana‑de‑açúcar

O objetivo deste trabalho foi avaliar o mapeamento de área de cana‑de‑açúcar por meio de série temporal, de seis anos de dados do índice de vegetação por diferença normalizada (NDVI), oriundos do sensor Vegetation, a bordo do satélite "système pour l'observation de la Terre" (SPOT). Três classes de cobertura do solo (cana‑de‑açúcar, pasto e floresta), do Estado de São Paulo, foram selecionadas como assinaturas espectro‑temporais de referência, que serviram como membros extremos ("endmembers") para classificação com o algoritmo "spectral angle mapper" (SAM). A partir desta classificação, o mapeamento da área de cana‑de‑açúcar foi realizado com uso de limiares na imagem-regra do SAM, gerados a partir dos valores dos espectros de referência. Os resultados mostram que o algoritmo SAM pode ser aplicado a séries de dados multitemporais de resolução moderada, o que permite eficiente mapeamento de alvo agrícola em escala mesorregional. Dados oficiais de áreas de cana‑de‑açúcar, para as microrregiões paulistas, apresentam boa correlação (r² = 0,8) com os dados obtidos pelo método avaliado. A aplicação do algoritmo SAM mostrou ser útil em análises temporais. As séries temporais de NDVI do sensor SPOT Vegetation podem ser utilizadas para mapeamento da área de cana‑de‑açúcar em baixa resolução.