Five skin and two oral biopsies from patients with South American pemphigus foliaceus (SAPF) were studied by electron and immunoelectron microscopy for the ultrastructural localization of bound immunoglobulin in epidermal and oral lesions. Electron microscopy showed the tonofilament-desmosome complex to be preserved in the various layers of the epidermis. Immunoglobulin was bound over the plasma membrane and permeated the desmosomal junctions both in the skin and oral mucosa, thus suggesting that pemphigus antibodies are attached to the glycocalyx. It appears that the initial injury in SAPF acantholysis involves the glycocalyx and that it might be caused by interaction with intercellular antibodies present in the patient's serum.
Five different antigens from Cysticercus cellulosae, a vesicular fluid, a saline and an alkaline total extracts, an escolex and membrane, were studied in the ELISA immunoenzymatic assay to demonstrate IgG antibodies in cerebrospinal fluid (CSF) and serum samples. For the 5 antigens the 20 micrograms/ml concentration was selected for polyvinyl plates sensitization for CSF and serum assays. The IgG fraction of a sheep anti-human IgG antiserum was labeled with horseradish peroxidase and revealed with hydrogen peroxide-5-aminosalicilic acid. For positive results 10 ELISA Units for sera and 5 EU for CSF were taken. A total of 182 serum samples and 115 CSF samples were tested. The ELISA sensitivity for sera were 88.6% for vesicular fluid, escolex and membrane; 85.7% for saline extract and 62.8% for alkaline extract. The ELISA sensitivity for CSF was 100% for vesicular fluid and saline total extract, 91.7% for membrane, 89.6% for excolex and 77.1% for alkaline. The ELISA specificity for sera was 100% to the 5 antigens studied; to the CSF was 100% for the alkaline and escolex and 98.5% for the other antigens. An ELISA geometric mean titer of sera and CSF was respectively the 121.1 and 27.3 for vesicular fluid, 111.6 and 31.1 for saline total extract, 30.3 and 5.3 for alkaline total extract, 63.2 and 14.8 for escolex and 69.1 and 18.8 for membrane antigen. ELISA was then compared to immunofluorescence, hemagglutination and complement fixation tests in CSF and sera. ELISA immunoenzymatic assay with saline total extract is recommended for the easy preparation and for the high quantity of antigens obtained, for the high sensitivity and great specificity for sera and CSF; we suggest that this test may be used as a substitute for immunofluorescence, hemagglutination and complement fixation in sera or CSF for the diagnosis of neurocysticercosis.
O presente trabalho teve como objetivos estimar a freqüência das formas músculo-cutánea e visceral da cisticercose em exames anátomo-patológicos e necrópsias realizados em Brasilia, Distrito Federal (estudo retrospectivo) e diagnosticar a cisticercose músculo-cutânea em pacientes residentes na mesma região geográfica (estudo prospectivo). Em 64.911 protocolos de exames anátomo-patológicos, o diagnóstico de cisticercose foi observado em 30 (0,05%), sendo que em 27 (90,0%) os cistos estavam nos tecidos músculo-cutâneo-mucoso, em 1 (3,3%) em gânglio e em 2 (6,7%) no sistema nervoso central. Entre aqueles com cistos nos tecidos músculo-cutâneo-mucoso 2 (7,4%) tinham cisticercos em língua. Em 1520 protocolos de necrópsia, encontraram-se 25 (1,6%) com diagnóstico de cisticercose, sendo: 24(96,0%) com neurocisticercose, seja isolada ou associada a outras formas da doença; e 2 (8,0%) com cisticercos em coração, 2 (8,0%) em músculo esquelético e 1 (4,0%) em fígado, seja isolados ou associados a outras localizações do parasito. Foram também examinados 1122 indivíduos, realizando-se em todos eles as reações sorológicas de imunofluorescência indireta e ELISA para cisticercose e a investigação radiológica de partes moles e crânio. Encontraram-se 59 (5,3%) com ambas reações sorológicas reagentes (10 entre eles com o diagnóstico de cisticercose confirmado por biópsias); e 32 (2,8%) com calcificações nas radiografias de partes moles e/ou crânio, mas apresentando ambas reações sorológicas não-reagentes. Entre os pacientes com os testes imunológicos reagentes, a neurocisticercose foi diagnosticada em 39 (66,1%), a cisticercose muscular em 25 (42,4%); a cutânea em 12 (20,3%); e a visceral em 2 (3,4%), sendo em 1 (1,7%) ovariana e em 1 (1,7%) miocárdica, pleural e renal. Os resultados permitem concluir que a forma músculo-cutânea é observada freqüentemente entre pacientes com cisticercose residentes em Brasília. A forma visceral também foi encontrada, com os cisticercos localizados em diferentes órgãos, sendo que os pacientes afetados não apresentavam as manifestações clínicas.
Estudo soroepidemiológico realizado em Brasília evidenciou a presença de infecção pelo Cysticercus cellulosae, detectada pelos testes imunoenzimáticos Elisa e imunofluorescência indireta, em 5,2% dos 1122 indivíduos avaliados. Entre os 120 líquidos cefalorraqueanos examinados, provenientes de pacientes que apresentaram sinais sugestivos de neurocisticercose, 16,7% foram reagentes. A prevalência da sorologia reagentefoi 20,4% no grupo de doentes com a hipótese diagnostica de cisticercose, 3,5% no grupo de seus familiares, 5,5% e 0,6% naqueles constituídos de pacientes ambulatoriais com cefaléia e epilepsia, respectivamente; e 0% no grupo controle. A cisticercose prevaleceu nas faixas etárias mais avançadas, nâo havendo predominância de sexo. No diagnóstico imunológico detectaram-se índices de positividoâe que variaram entre os grupos naturais das diversas regiões do país, sendo encontrados 8,1% de indivíduos sororreagentes no Sudeste, 5,8% no Nordeste, 5,3% no Centro-Oeste e 3,5% no Sul do país. Dos fatores epidemiológicos, a ausência de condições sanitárias nas residências, o maior contato com suínos, e o uso de água de rio constituíram os maiores riscos para contrair a moléstia, sendo seu risco relativo de 3,1, 2,2 e 1,8, respectivamente.
IgM antibodies against Cysticercus cellulosae in the cerebrospinal fluid (CSF) was demonstrated by ELISA immunoenzymatic assay in neurocysticercosis. CSF samples of 41 patients were analyzed for this purpose. Diagnosis was neurocysticercosis in 26 and neurosyphilis in 5; abnormalities were not registered in the other 10 cases. Neurosyphilis samples and no-abnormalities samples were considered as control groups. ELISA IgM assay for cysticercosis was negative in all CSF samples of control groups and it was positive in 12 of the 26 CSF samples of the neurocysticercosis group (46.2%). Titers ranged from 4 till 32. Positive results were no more obtained after previous treatment of CSF samples by 2-mercaptoethanol. ELISA IgM and IgG titers were compared. IgM titers wee higher than IgG titers in two cases. Results obtained were compared to those found through complement fixation, immunofluorescence and hemagglutination tests for the diagnosis of neurocysticercosis.
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