Familial hypercholesterolemia: Is there a role for PCSK9 and thrombin generation?

INTRODUCTION: Familial hypercholesterolemia (FH) is an autosomal dominant genetic disease. The prevalence of FH has previously been reported as 1 in 500 in the general population. This study aimed to evaluate the proprotein convertase subtilisin/kexin 9 (PCSK9) levels, lipid profile and thrombin generation in FH patients undergoing treatment or not. METHODS: Eighty individuals with FH were selected and distributed in 2 groups: individuals treated with statins alone or conjugate therapy (statin + ezetimibe) (T = 53) and those non treated (NT = 27). PCSK9 levels were determined by ELISA, the lipid profile by colorimetric enzyme method and thrombin generation assay (TGA) by CAT method. RESULTS: Individuals treated with conjugate therapy (statin + ezetimibe) showed a significant reduction in the levels of total cholesterol (TC) low density lipoprotein cholesterol (LDLc) and in the potential for thrombin generation (ETP with low and high concentration of tissue factor), compared to the treated individuals with monotherapy (statins). PCSK9 was positively correlated with increased levels of TC, LDLc and triglycerides, while TGA parameters were positively correlated with PCSK9 and lipid profile. CONCLUSION: PCSK9 levels appear to be associated with components of the lipid and hemostatic profiles, in addition to being influenced by age. In general, our findings suggest that combined therapy for the treatment of FH is associated with a significant improvement in both lipid and hemostatic profiles assessed by TGA, suggesting a reduction in atherogenic and thrombogenic risks and, therefore, more promising compared to the use of statin monotherapy.

Induction and growth pattern of callus from Piper permucronatum leaves / Indução e padrão de crescimento de calos de folhas de Piper permucronatum

ABSTRACT Piper permucronatum is a perennial shrub, a medicinal plant native to the Amazon Rainforest. Traditionally, the tea of its leaves is used to combat menstrual and intestinal cramps, stomach pain, digestive problems, diarrhea, hemorrhage, and nausea. Its leaf’s essential oil is effective against Aedes aegypti larvae; its flavones and flavanones have a fungicidal effect against Clamidosporium cladosporioides and C. sphaerospermum; its hexanic extract is effective against Leishmania amazonensis. The objective of this study was to provide a protocol for callus induction from P. permucronatum leaves and an identification of the callus growth pattern, focusing on the deceleration phase, when the callus cells must be subcultured into liquid medium in order to produce a cell suspension cultures. Leaf explants were inoculated in a solid MS medium supplemented with factorial combinations of 2,4-D, BA, NAA and GA3. Callus formation was evaluated weekly until the 49th day. Subsequently, new explants were inoculated at the hormonal combination that resulted in the highest callus cell proliferation and, every seven days during a period of 70 days, samples were dried and weighed to determine the callus growth pattern. NAA and GA3 were not effective for callus induction. Combinations of 2,4-D and BA resulted in callus induction and proliferation. The highest percentage of callus induction was observed with the combination of 4.52 µM 2,4-D and 4.44 µM BA. The calluses thereby produced were friable and whitish. The callus growth pattern followed a sigmoid shape. The deceleration phase started on the 56th day of culture.

RESUMO Indução e padrão de crescimento de calos de folhas de Piper permucronatum. Piper permucronatum é um arbusto perene, uma planta medicinal native da Floresta Amazônica. Tradicionalmente, o chá de suas folhas é usado em casos de cólicas menstruais e intestinais, dores de estômago, problemas digestivos, diarreia, hemorragia e náusea. O óleo essencial das folhas é efetivo contra a larva de Aedes aegypti; suas flavonas e flavanonas têm efeito fungicida contra Clamidosporium cladosporioides e C. sphaerospermum; seu extrato hexânico é efetivo contra Leishmania amazonensis. O objetivo deste trabalho foi determinar um protocolo para indução de calos em folhas de P. permucronatum e identificar o padrão de crescimento dos calos, com foco na fase de desaceleração, quando as células de calo devem ser subcultivadas em meio líquido para produzir culturas de células em suspensão. Explantes foliares foram inoculados em meio MS sólido suplementado com combinações fatoriais de 2,4-D, BAP, ANA e GA3. A formação de calos foi avaliada semanalmente até o 49º dia. Posteriormente, novos explantes foram inoculados na combinação hormonal que resultou na maior proliferação de células de calo e, a cada sete dias durante 70 dias, amostras foram secas e pesadas para determinar o padrão de crescimento dos calos. ANA e GA3 não foram efetivas para a indução de calos. Combinações de 2,4-D e BAP resultaram em indução e proliferação de calos. A maior porcentagem de indução de calos foi observada com a combinação de 4,52 µM de 2,4-D e 4,44 µM de BAP. Os calos produzidos eram friáveis e esbranquiçados. O crescimento dos calos seguiu um padrão sigmoide. A fase de desaceleração iniciou no 56º dia de cultivo.

Effects of periodontal therapy on white blood cell count and levels of transforming growth factor beta in serum of subjects with severe periodontitis.

This study aimed to investigate the effects of nonsurgical periodontal therapy on white blood cell (WBC) count and levels of transforming growth factor beta (TGF—β) in serum from subjects with severe periodontitis. Serum from 28 subjects with periodontitis (mean age: 34.36±6.24; 32% men) and 27 healthy controls (mean age: 33.18±6.42; 33% men) were collected prior to therapy. Blood samples were obtained from 23 subjects who completed therapy (9—12 months). A well—controlled periodontal treatment protocol was established in three stages: mechanical periodontal therapy (scaling and root planning), reinstrumentation of dental sites, and supportive periodontal therapy. Periodontal and systemic parameters such as the total number of WBCs and TGF—β levels, accessed by enzyme—linked immunosorbent assay (ELISA), were included. After therapy, all clinical periodontal parameters decreased (p<0.0001). There were no statistical differences in WBC count between experimental and control groups before or after therapy. However, after therapy, the mean value of lymphocytes in patients with localized aggressive periodontitis (LAgP) was statistically higher than that of patients with generalized chronic periodontitis (GCP) (p<0.0357). Additionally, TGF—β levels in LAgP and GCP patients were higher compared to controls before therapy (p<0.05 and p<0.01, respectively). In LAgP patients, periodontal therapy was associated with increased number of lymphocytes.

Callogenesis in leaves of Kalanchoe pinnata Lam. by 2, 4-D and BA action / Calogênese em folhas de Kalanchoe pinnata Lam. pela ação de 2, 4-D e BAP

The Kalanchoe pinnata Lam. is a bush species of the Crassulaceae that is distinguished by its important medicinal properties. Its leaves are used as cataplasm to treat headaches and wounds. There is evidence for a hypotensive and anti-inflammatory effect. Techniques of plant tissue culture have been applied to plant species that produce substances likely to be explored in pharmacology, cell suspension being the main technique. At the industrial level, this method utilizes bioreactors in order to produce secondary metabolites on a large scale. The objective of this study was to evaluate the effects of in vitro combinations of 2,4-dichlorophenoxiacetic acid (2,4-D) and benzylaminopurine (BA) on callus induction in leaf explants of K. pinnata. Leaf fragments were inoculated in MS medium supplemented with 3.0% sucrose, 0.8% agar and factorial combinations of 2,4-D (0.00, 4.52, 9.06, 18.12 µM) and BA (0.00, 4.44, 8.88, 17.76 µM). The cultures were kept in the darkness at 24±2ºC for 50 days. The percentage of callus induction and the area of explants covered by callus cells were evaluated. In the absence of growth regulators, callus induction did not occur, with necrosis of all explants. The highest percentage of callus induction was 100%, obtained with the combination of 9.06 µM 2,4-D and 8.88 µM BA, but the calluses covered only 25% of the leaf area. The most efficient combination was 4.52 µM 2,4-D and 8.88 µM BA, resulting in 91% callus induction with 50 to 100% of the explants being covered by callus cells.

Kalanchoe pinnata Lam. é uma espécie arbustiva da família Crassulaceae que apresenta interessantes propriedades medicinais. Suas folhas são utilizadas em cataplasma para tratar enxaqueca e ferimentos. Há evidência de seu efeito como hipotensiva e anti-inflamatória. Técnicas de cultura de tecidos vegetais têm sido aplicadas para espécies que possuem substâncias passíveis de exploração na farmacologia, sendo a suspensão celular a principal técnica utilizada. A nível industrial, este método utiliza biorreatores para produzir metabólitos secundários em larga escala. Este estudo teve como objetivo avaliar os efeitos in vitro de combinações do ácido 2,4-diclorofenoxiacético (2,4-D) e de benzilaminopurina (BAP) na indução de calos em explantes foliares de K. pinnata. Fragmentos foliares foram inoculados em meio MS contendo 3% de sacarose, 0,8% de ágar e combinações fatoriais de 2,4-D (0,00; 4,52; 9,06 e 18,12 µM) e BAP (0,00; 4,44; 8,88 e 17,76 µM). Os cultivos foram mantidos no escuro, a 24±2ºC por 50 dias. A porcentagem de indução de calos e a área dos explantes coberta por células de calos foram avaliadas. Na ausência de reguladores de crescimento não ocorreu indução de calos, com necrose de todos os explantes. A porcentagem mais alta de indução de calos foi de 100%, obtida com a combinação de 9,06 µM de 2,4-D e 8,88 µM de BAP, mas estes calos cobriram apenas 25% da área foliar. A combinação mais eficiente foi de 9,06 µM de 2,4-D e 8,88 µM de BAP, que resultou em 91% de indução e 50 a 100% da área dos explantes coberta por células de calos.

[The subject repositories of strategy of the Open Access initiative]. / Los repositorios temáticos en la estrategia de la iniciativa Open Access.

The subject repositories are defined as a set of digital objects resulting from the research related to a specific disciplinary field and occupy a still restricted space in the discussion agenda of the Free Access Movement when compared to amplitude reached in the discussion of Institutional Repositories. Although the Subject Repository comes to prominence in the field, especially for the success of initiatives such as the arXiv, PubMed and E-prints, the literature on the subject is recognized as very limited. Despite its roots in the Library and Information Science, and focus on the management of disciplinary collections (subject area literature), there is little information available about the development and management of subject repositories. The following text seeks to make a brief summary on the topic as a way to present the potential to develop subject repositories in order to strengthen the initiative of open access.

Interleukin-6 (G-174C) and tumour necrosis factor-alpha (G-308A) gene polymorphisms in geriatric patients with chronic periodontitis.

BACKGROUND AND OBJECTIVE: Periodontitis is a chronic inflammatory disease, and genetic factors may have an important role in its severity. Polymorphisms in the promoter regions of the interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha) genes have been reported to cause changes in the production of these cytokines. The aim of this study was to evaluate the possible role of IL-6 (G-174C) and tumour necrosis factor (G-308A) polymorphisms, in the severity of chronic periodontitis in an elderly population. MATERIALS AND METHODS: In this study, a group of 65 elderly women, comprising 17 patients with moderate chronic periodontitis, 21 with severe chronic periodontitis and 27 healthy patients were selected. DNA was isolated from all subjects, and polymerase chain reaction was used to study the IL-6 and TNF-alpha gene polymorphisms. RESULTS: The results of this study showed a significant difference in the allele and genotype frequencies of IL-6 gene polymorphism between patients with periodontal disease and controls. Subjects carrying the G/G genotype of IL-6 were most severely affected by periodontitis. The TNF-alpha gene polymorphism showed no association with chronic periodontitis between patients and controls. CONCLUSION: The results suggest that the IL-6 gene polymorphism may be associated with chronic periodontitis, and that TNF-alpha gene polymorphism may not be involved in the progression of chronic periodontitis in the population of elderly Brazilian women.

Assessing and analysing contamination of a dairy products processing plant by Staphylococcus aureus using antibiotic resistance and PFGE.

A dairy product processing plant was studied for 2.5 years to examine contamination with Staphylococcus aureus and try to correlate the source of contamination. Cultures were submitted to an antibiotic susceptibility test (AST) and characterised by Pulsed-field Gel Electrophoresis (PFGE) analysis. Results showed that 35.2% (19/51) of food handlers were asymptomatic carriers of S. aureus, and that 90.4% (19/21) of raw milk sampled was contaminated. Staphylococcus aureus was isolated from only 10 samples among more than 3200 investigated dairy products. No S. aureus contamination was found on machinery. The AST analysis demonstrated sensitivity of tested S. aureus to oxacillin, cephalothin, vancomycin, gentamicin, and sulfamethoxazole/trimethoprim. AST analysis generated eight different phenotypic profiles, but did not allow us to identify the source of contamination in seven of ten final products. PFGE analysis proved to be a sensitive method as it generated 42 different DNA banding profiles among the 48 S. aureus investigated, demonstrating a lack of predominance of endemic strains in the plant, contrary to suggestions raised by antibiotic resistance typing. Based on PFGE genotyping, S. aureus strains isolated from four contaminated final products were similar to four S. aureus isolated from raw milk. Five final products contained S. aureus different from all other strains collected, and one showed similarity to a strain isolated from a food handler. These results suggest contamination by raw milk as the main source of contamination of the final dairy products.

[Hypogammaglobulinemia as risk factor for Cryptococcus neoformans infection: report of 2 cases]. / Hipogamaglobulinemia como fator de risco para infecção por Cryptococcus neoformans: a propósito de dois casos.

Two HIV-seronegative patients with cryptococcal disease refractory to conventional antifungal therapy were submitted to an evaluation of the immune system. Hypogammaglobulinemia was found in both and associated with abnormal function of cell-mediated immunity. Hypogammaglobulinemia is considered as a possible predisposing factor for cryptococcal infection. The importance of the antibodies on the control of Cryptococcus neoformans infection is discussed.

Reproducibility of alkaline antigens of Leishmania major-like and Leishmania (V.) braziliensis evaluated by IgG-ELISA. Comparison of antigens added of a protein inhibitor (PMSF) or not.

This paper deals with the analysis of 10 batches of L.major-like and L. (V.) braziliensis antigens added or not of a proteases inhibitor evaluated by means of an IgG-ELISA on three consecutive days using positive standard sera from patients with diagnosis of American Leishmaniasis previously tested for the presence of IgG antibodies by means of ELISA. The statistical analysis showed that for L. (V.) braziliensis the PMSF-containing antigen did not show any difference among batches or days of testing; the L. (V.) braziliensis antigen without PMSF showed statistical significance for differences among batches and a two-way ANOVA showed significant differences between antigens. L.major-like antigen prepared with or without PMSF showed differences among batches; all 3 days of testing displayed differences for the PMSF antigen but only for days 1 and 2 for the antigen without inhibitor. A two-way ANOVA showed differences among batches of the antigens but not for antigens with and without the protein inhibitor. According to the statistical analysis the L.major-like antigen added or not of PMSF has shown that it is the choice antigen for mucocutaneous leishmaniasis serology.

Evaluation of Leishmania antigens preparation and storage for use in enzyme immunoassays

The influence of time and temperature on the storage of an alkaline antigen of L. major-like and L.(V.) braziliensis promastigotes added or not of a proteases inhibitor (PMSF) was evaluated by means of an IgG-ELISA. Antibodies in assays using L. major-like antigen stored at -20 degrees C for 6 months had a statistically lower geometric mean titer (GMT) and different 95% confidence interval limits (CL) than antigens stored otherwise, as assessed by the [quot ]t[quot ] statistic. The PMSFL. major-like antigen after storage for 6 months at a temperature of 4 degrees C had the same GMT and 95% CL displayed at time zero as well as when storage for 4 and 6 months at -20 degrees C. Significant differences were not found when L.(V.) braziliensis antigens were stored at times and temperatures mentioned; the PMSF antigen stored for 2 months at -70 degrees C resulted in a lower serum GMT and 95% CL than any other, as assessed by the [quot ]t[quot ] statistic. Antigen performance did not show any statistical difference associated to the addition of PMSF within the same species; the largest difference between antigens was that between PMSF-L. (V.) braziliensis and L. major-like without PMSF.

A influência do tempo e temperatura de estocagem de antígenos alcalinos de promastigotas de L. major-like e L. (V.) braziliensis adicionados ou não de um inibidor de proteases foi avaliada por meio de reações de IgG-ELISA. A reação que empregava o antígeno de L. major-like estocado por 6 meses a -20oC mostrou que médias geométricas dos títulos (MGT)e intervalos de confiança 95% (IC 95%) eram estatisticamente inferiores àquelas obtidas com antígenos estocados em outros intervalos de tempo, medido pela estatística "t". O antígeno PMSF-L. major-like depois de 6 meses de estocagem à temperatura de 4oC tinha a mesma MGT e IC 95% do tempo zero assim como quando ele foi estocado a -20oC por 4 e 6 meses. Não foram observadas diferenças estatisticamente diferentes com os antígenos de L. (V.) braziliensis estocados nas mesmas condições de tempo e temperatura exceto o antígeno PMSF estocado por 2 meses a -70oC que apresentou MGT e IC 95% inferiores a quaisquer outras como aferido pela estatísitca "t". Quando comparados os desempenhos dos antígenos não houve direrenças estatisticamente significantes entre a adição ou não de PMSF para qualquer dos parasitas. A análise do cruzamento entre antígenos mostrou que a maior diferença netre eles foi a do contraste entre L. (V.) braziliensis adicionado de PMSF e L. major-like sem adição de PMSF.

Evaluation of Leishmania antigens preparation and storage for use in enzyme immunoassays.

The influence of time and temperature on the storage of an alkaline antigen of L. major-like and L.(V.) braziliensis promastigotes added or not of a proteases inhibitor (PMSF) was evaluated by means of an IgG-ELISA. Antibodies in assays using L. major-like antigen stored at -20 degrees C for 6 months had a statistically lower geometric mean titer (GMT) and different 95% confidence interval limits (CL) than antigens stored otherwise, as assessed by the "t" statistic. The PMSFL. major-like antigen after storage for 6 months at a temperature of 4 degrees C had the same GMT and 95% CL displayed at time zero as well as when storage for 4 and 6 months at -20 degrees C. Significant differences were not found when L.(V.) braziliensis antigens were stored at times and temperatures mentioned; the PMSF antigen stored for 2 months at -70 degrees C resulted in a lower serum GMT and 95% CL than any other, as assessed by the "t" statistic. Antigen performance did not show any statistical difference associated to the addition of PMSF within the same species; the largest difference between antigens was that between PMSF-L. (V.) braziliensis and L. major-like without PMSF.

[Evaluation of the possible influence of Montenegro's intradermal reaction on a serological assay for the diagnosis of American cutaneous leishmaniasis]. / Avaliação de eventual influência da intradermorreação de Montenegro sobre prova sorológica para o diagnóstico da Leishmaniose tegumentar americana.

Influence of Montenegro skin test for American tegumentar leishmaniasis was evaluated to verify possible interference in serological diagnosis for this disease, performed by immunoenzymatic assay (ELISA). If this interference could occur, it would hamper scientific, epidemiological and patient care; happily we did not find any interference on serological diagnosis by performance of skin testing.

Rendimiento diagnóstico de la inmunodeficiencia y el ensayo inmunoenzimático en sueros de pacientes con leishmaniasis del norte y del nordeste de Brasil / Diagnostic performance indices for immunofluorescent tests and enzyme immunoassays of leshmaniasis sera from northern and northeastern Brazil

Se examinó un total de 341 muestras de suero para detectar anticuerpos IgA, IgG e IgM anti-Leishmania mediante pruebas de inmunofluorescencia (IF) y ensayos inmunoenzimáticos (ELISA). En conjunto, 292 de los sueros pertenecían a pacientes con diagnósticos clínicos y parasitológicos de leishmaniasis cutánea (resultados positivos en la impronta de lesiones o en la prueba cutánea de Montenegro) y 49, a controles de la misma población. Los índices de rendimiento diagnóstico de los ELISA-IgG e IgM fueron de utilidad diagnóstica y el valor predictivo positivo del ELISA-IgG fue de 94,6. La especificidad de la prueba de IF-IgA fue notablemente alta (100), pero su sensibilidad fue muy baja.

Rendimiento diagnóstico de la inmunodeficiencia y el ensayo inmunoenzimático en sueros de pacientes con leishmaniasis del norte y del nordeste de Brasil / Diagnostic performance indices for immunofluorescent tests and enzyme immunoassays of leshmaniasis sera from northern and northeastern Brazil

Se examinó un total de 341 muestras de suero para detectar anticuerpos IgA, IgG e IgM anti-Leishmania mediante pruebas de inmunofluorescencia (IF) y ensayos inmunoenzimáticos (ELISA). En conjunto, 292 de los sueros pertenecían a pacientes con diagnósticos clínicos y parasitológicos de leishmaniasis cutánea (resultados positivos en la impronta de lesiones o en la prueba cutánea de Montenegro) y 49, a controles de la misma población. Los índices de rendimiento diagnóstico de los ELISA-IgG e IgM fueron de utilidad diagnóstica y el valor predictivo positivo del ELISA-IgG fue de 94,6

. La especificidad de la prueba de IF-IgA fue notablemente alta (100

), pero su sensibilidad fue muy baja

Rendimiento diagnostico de la inmunodeficiencia y el ensayo inmunoenzimatico en sueros de pacientes con leishmaniasis del norte y del nordeste de Brasil / Diagnostic performance índices for immunofluorescent tests and enzyme immunoassays of leishmaniasis sera from northern and northeastern Brazil

Se examino un total de 341 muestras de suero para detectar anticuerpos IgA, IgG e IgM anti-Leishmania mediante pruebas de inmunofluorescencia (IF) y ensayos inmunoenzimaticos (ELISA). En conjunto, 292 de los sueros pertenecian a pacientes con diagnosticos clinicos y parasitologicos de leishmaniasis cutanea (resultados positivos en la impronta de lesiones o en la prueba cutanea de Montenegro) y 49, a controles de la misma poblacion. Los índices de rendimiento diagnostico de los ELISA-IgG e IgM fueron de utilidad diagnostica y el valor predictivo positivo del ELISA-IgG fue de 94,6. . La especificidad de la prueba de IF-IgA fue notablemente alta (100. ), pero su sensibilidad fue muy baja

Comparison on the performance of Leishmania major-like and Leishmania braziliensis braziliensis as antigen for New World leishmaniasis IgG-immunofluorescence test.

The performance of an antigen of L. major-like promastigotes for the serological diagnosis of mucocutaneous leishmaniasis in the IgG-immunofluorescent test was compared to that of an antigen of L.braziliensis braziliensis. Each antigen was used to test two hundred and twenty-four sera of etiologies such as mucocutaneous leishmaniasis, deep mycoses, toxoplasmosis, malaria. Chagas' disease, visceral leishmaniasis, anti-nuclear factor, schistosomiasis, rheumatoid factor and normal controls. Agreement between responses to each antigen was high: 77.2% of leishmaniases sera agreed on a positive or a negative result to both antigens and 91.1% of control sera. Cross reactivity was restricted to Chagas' disease sera, visceral leishmaniasis, anti-nuclear factor and paracoccidioidomycosis. The quantitative response of leishmaniasis and Chagas' disease sera to both antigens was evaluated by a linear regression; although the y-intercept and the slope were different for each antigen, neither was better than the other in the disclosure of anti-Leishmania antibodies. In the case of Chagas' disease sera the L.major-like antigen was better than L.b.braziliensis' to disclose cross-reacting antibodies.

Performance indexes of a dot-enzyme-linked immunosorbent assay (dot-ELISA) and an enzyme-linked immunosorbent assay (IgG-ELISA) for field surveys of New World leishmaniasis.

Diagnostic performance indexes of sensitivity, specificity, positive predictive value and efficiency were determined for dot-ELISA and IgG-ELISA tests in 340 leishmaniasis sera. Sensitivity of the dot-ELISA was significantly lower than IgG-ELISA's; the two tests had indexes of specificity and positive predictive value of the same magnitude. Seventy-eight sera gave a negative dot-ELISA test result and a positive IgG-ELISA test result. When sera were classified according to different criteria as how to interpret this diversity, the kappa statistic did not corroborate the classification indicating that the two tests display a substantial strength of agreement. The results presented indicate that performance indexes accrued in a survey where variables are well known may be extrapolated to other population studies if the disease presents itself as highly prevalent (due to a selection bias or not) and may be expected to discriminate a disease status among test positives.

Diagnostic performance indices for immunofluorescent tests and enzyme immunoassays of leishmaniasis sera from northern and north-eastern Brazil.

A total of 341 sera were screened for anti-Leishmania IgA, IgG, and IgM antibodies by immunofluorescent (IF) tests and enzyme immunoassay (ELISA). Altogether, 292 of the sera originated from patients with clinically as well as parasitologically diagnosed (positive lesion imprint or the Montenegro skin test) cutaneous leishmaniasis; 49 of the sera were from controls from the same base population. In terms of diagnostic performance, the ELISAs for IgG and IgM yielded indices of diagnostic utility, and the positive predictive value for the IgG-ELISA was 94.6%. A remarkably high specificity (100%) was obtained with the IgA-IF test, but its sensitivity was very low.