The association of autoimmune thyroiditis and non-functional adrenal incidentalomas with insulin resistance

Objective Patients with incidental nonfunctioning adrenal adenoma are associated with increased risk of obesity, impaired glucose tolerance and dyslipidemia. We aimed to investigate the relationship between thyroid function, serum lipids and insulin resistance in patients with nonfunctioning adrenal incidentaloma. Subjects and methods Forty patients who had diagnosed as adrenal incidentaloma (AI) in our department were included in the study. Serum free triiodothyronine (fT3), free thyroxine (fT4), thyroid stimulating hormone (TSH), anti-thyroperoxidase antibody (anti-TPO Ab) and anti-thyroglobulin antibody (anti-Tg Ab), lipid profile, hs-CRP, fasting insulin levels were measured and insulin resistance calculated by HOMA-IR. Thyroid volume (TV) was assessed. Results None of the patients showed specific signs and symptoms of hormone excess. TV, TSH and fT3 levels in the patient and control groups did not differ significantly (p > 0.05). The serum fT4, anti-TG Ab, anti-TPO Ab levels in the patient group were significantly higher than in the control group (p = 0.013, p < 0.0001, p = 0.016 respectively). The HOMA-IR, hs-CRP and HDL cholesterol levels in the AI patients were significantly higher than the control group (p = 0.034, p = 0.041, p = 0.002, respectively). Statistically significant relationship was found between HOMA-IR and thyroid volume (r = 0.373, p = 0.018), fT4 (r = 0.382, p = 0.015), hs-CRP (r = 0.512, p = 0.001), HDL cholesterol (r = 0,351 p = 0.026) in AI patients. There were significant correlation between anti-TG Ab, anti-TPO Ab and TSH levels in AI patients (r = 0.431 p = 0.006, r = 0.402 p = 0.012). Conclusions Patients with nonfunctioning adrenal incidentaloma have several metabolic disturbances. At the same time autoimmune thyroid disorders are more frequent in nonfunctioning adrenal incidentaloma patient so that thyroid functions must be evaluated in those patients. Arch Endocrinol Metab. 2015;59(1):42-6 .

Elevada frecuencia de enfermedad tiroidea funcional en embarazadas chilenas sin antecedentes de patología tiroidea utilizando el estándar de TSH internacional / Frequency of subclinical thyroid problems among women during the first trimester of pregnancy

Background: Thyroid hormones play an important role in fetal neural and cognitive development. Therefore thyroid abnormalities should be detected and treated early during pregnancy. Aim: To assess the frequency and risk factors for functional thyroid disorders during the first trimester of pregnancy. Material and Methods: A blood sample was obtained from women during their first trimester of pregnancy, consulting in a prenatal care facility. Women with known thyroid diseases were excluded from the study. Thyroid stimulating hormone (TSH), total thyroxine (T4) and free thyroxine (fT4) were measured by electrochemoluminiscence. Antithyroid peroxidase antibodies (anti TPO) were measured by enzyme immunoassay. Results: Five hundred and ten women aged 25.7 ± 6.6 years were assessed. The frequency of clinical hypothyroidism was 0.6%, subclinical hypothyroidism 35.3% and clinical hyperthyroidism 1%. Five percent of women with hypothyroidism and 3.5% of euthyroid women had positive anti TPO antibodies. There was no association between the frequency of thyroid diseases and risk factors for thyroid diseases. Conclusions: There is a high frequency of subclinical thyroid diseases among women consulting in this prenatal care clinic.

Quantificação de anticorpos antitireoperoxidase e antitireoglobulina, tireotrofina e tiroxina livre em gestantes normais / Quantification of antithyroperoxidase and antithyroglobulin antibodies, and thyrotrophin and free thyroxine in normal pregnant women

OBJETIVO: verificar a presença de anticorpos antitireoperoxidase (anti-TPO) e antitireoglobulina (anti-TG) e as concentrações plasmáticas de tireotrofina (TSH) e tiroxina livre (T4L) em gestantes normais. MÉTODOS: estudo transversal realizado no ambulatório de Pré-Natal da Santa Casa de Misericórdia de São Paulo (SP), no período de janeiro de 2003 a setembro de 2004. Foram incluídas 127 grávidas, residentes em São Paulo, com idades entre 14 e 44 anos e idade gestacional igual ou superior a 16 semanas, determinada por ultra-sonografia realizada antes da 20ª semana de gestação. Foram excluídas gestantes em uso de medicamentos ou história de tireoidopatia. Foram quantificados os anticorpos antitireoperoxidase e antitireoglobulina por imunoensaio quimioluminescente. A técnica de imunofluorimetria por tempo resolvido foi empregada para a determinação de tireotrofina e tiroxina livre. Para análise dos resultados, aplicou-se o teste t de Student, com significância de 5 por cento. RESULTADOS: a freqüência dos anticorpos antitireoidianos foi 12,6 por cento (8,6 por cento de anticorpos anti-TPO, 4,6 por cento de anticorpos anti-TG). A média das concentrações de TSH foi de 2,1±1,0 µU/mL e a média de T4L foi de 0,9±0,5 ng/dL. Observou-se alteração da função tireoidiana em dez gestantes (8 por cento). Três delas tiveram diagnóstico de hipotireoidismo: uma na forma clínica da doença, com TSH aumentado e T4L diminuído; duas na forma subclínica, com TSH aumentado e T4L normal. Cinco mostraram valores de TSH diminuídos e de T4L aumentados, compatíveis com hipertireoidismo clínico, e duas tiveram diagnosticado hipertireoidismo subclínico, apenas com concentrações de TSH diminuídas. CONCLUSÕES: a freqüência de anticorpos antitireoidianos foi de 12,6 por cento em gestantes, sendo os anticorpos antitireoperoxidase predominantes sobre os anticorpos antitireoglobulina. Verificaram-se disfunções da tireóide em 8 por cento dos casos, com alterações...

PURPOSE: to quantify the presence of antithyroperoxidase (anti-TPO) and antithyroglobulin (anti-TG) antibodies, and the plasmatic concentrations of thyrotropin (TSH) and free thyroxine (FT4) in normal pregnant women. METHODS: a hundred twenty-seven pregnant women, residing in São Paulo, aged from 14 to 44 years old and gestational age > 16 weeks, determined by ultrasound performed before the 20th week of pregnancy were included in a transversal study performed in the prenatal clinic of Santa Casa de Misericórdia de São Paulo, from January 2003 to September 2004. Pregnant women using medicines or with thyroidopathy history were excluded. Antithyroperoxidase and antithyroglobulin antibodies were quantified by chemiluminescence immunoassay. The immunofluorimetry technique by time-resolved was used for determining the thyrotrophin and free thyroxine. The Student's t test, with significance of 5 percent, was used for analyzing the results. RESULTS: the frequency of antithyroid antibodies was 12.6 percent (8.6 percent of anti-TPO antibodies, and 4.6 percent of anti-TG antibodies). The average of TSH concentrations was 2.13±1.0 µU/ml, and the average of T4L was 0.9±0.5 ng/dl. It was observed alteration of the thyroid function in ten pregnant women (8 percent). Three of them had diagnosis of hypothyroidism: one in the clinical form of the disease, with increased TSH and decreased FT4; two in the subclinical form with increased TSH and normal FT4. Five presented decreased TSH and increased FT4, consistent with clinical hyperthyroidism and two were diagnosed with subclinical hyperthyroidism, with decreased TSH concentrations only. CONCLUSIONS: the frequency of antithyroid antibodies was 12.6 percent in pregnant women, the antithyroperoxidase antibodies being predominant over the antithyroglobulin antibodies. It was observed some thyroid dysfunction in 8 percent of the cases with alterations of TSH and/or T4L.

Moyamoya disease associated with positive findings for rheumatoid factor and myeloperoxidase-anti-neutrophil cytoplasmic antibody / La enfermedad de moyamoya asociada con hallazgos positivos en relación con el factor reumatoideo y los anticuerpos citoplasmáticos antineutrófilos mieloperoxidasa

A 39-year old female suddenly fell into a state of unconsciousness. She had no significantpast medical history. A computed tomography scan of the head demonstrated a massive left putaminal haemorrhage with a ventricular perforation, low density areas in the right frontal lobe, corona radiata and occipital lobe. A single emergency burr hole drainage of the haematoma was performed. Bilateral common carotid arteriograms showed stenosis of the right internal carotid artery and a complete obstruction of left internal carotid artery which were both accompanied by moyamoya vessels. The biochemical studies indicated serological positive findings for RF and MPO-ANCA. She was transferred to another hospital for nursing care in a vegetative state on the 163nd hospital day. This case indicates that immunological factors, inflammation or vasculitis might have possibly been associated with the development of either an obstruction or stenosis of the intracranial internal carotid arteries.

Auto-imunidade ANCA (Anticorpo Anti-Citoplasma de Neutrófilos) positiva induzida por propiltiouracil: relato de caso e revisão da literatura / Antineutrophil Cytoplasmic Antibody (Anca)-Associated autoimmune disease induced by propylthiouracil

A terapêutica com drogas antitireoidianas pode ser acompanhada de efeitos colaterais. Propiltiouracil (PTU) pode induzir vasculites anticorpo anti-citoplasma de neutrófilos (ANCA) positivas, na maioria das vezes relacionadas ao subtipo mieloperoxidase (ANCA-MPO). O nosso objetivo é relatar o caso de uma paciente com doença de Graves que desenvolveu auto-imunidade induzida por PTU, com manifestações cutâneas, pulmonares e renais, associadas à positividade do ANCA. O exame anátomo-patológico pulmonar revelou hemorragia difusa e a biópsia renal demonstrou glomeruloesclerose segmentar e focal. Foi tratada com pulsoterapia com corticóides e ciclofosfamida, com boa evolução clínica. Este caso enfatiza a necessidade de detecção e tratamento precoce deste efeito adverso relativamente raro do PTU.

Antithyroid drugs sometimes cause severe complications. Propylthiouracil (PTU) can be associated to ANCA positive vasculitis, most often related to myeloperoxidase subtype (ANCA-MPO). Our objective is to describe a female patient with Graves' disease, who developed PTU induced-autoimmune disease, with cutaneous, pulmonary, and renal lesions, associated with ANCA. Histopathological examination revealed diffuse pulmonary hemorrhage, and focal segmental glomerulosclerosis at the kidney biopsy. She was treated with systemic corticosteroid therapy and cyclophosphamide, with clinical improvement. This case highlights the need for greater awareness of this relatively rare adverse effect of propylthiouracil.

Anti-myeloperoxidase antibodies in patients with microscopic polyangiitis.

BACKGROUND & OBJECTIVES: Anti-neutrophil cytoplasmic antibodies in active necrotizing and crescentic glomerulonephritis are associated with systemic vasculitides like Wegener's granulomatosis, Microscopic polyangitiis and Churg Strauss Syndrome. This study shows the incidence of ANCA with specificities to Myeloperoxidase and Proteinase3 in MPA cases and gives the correlation of ANCA with Birmingham Vasculitis Activity Score. MATERIAL & METHODS: Eighteen cases of MPA were diagnosed as per Chapel Hill Consensus Criteria. ANCA was detected by indirect immunofluorescence microscopy using fluorescence and Confocal Laser Scanning Microscopes. Anti-MPO and anti-PR3 were identified by commercial ELISAs and anti-MPO subclass and IgG isotypes were also detected. RESULTS: MPA patients showed a male preponderance with BVAS ranging from 17-30. Systemic involvement was seen in 88.9%, lower respiratory tract involvement in 77.8% and upper respiratory tract in only 33.3% cases. All these patients had perinuclear pattern on IIF, where titers ranged from 80-640 and ELISA showed anti-MPO; values ranging from 20-80 units/ml. IIF and ELISA showed a good correlation (r=0.77). Two patients having FPGN had dual specificities and had both anti-MPO and anti-PR3 which could be picked up only by ELISA. A good correlation (r=0.78) was observed between BVAS and ANCA levels as well. IgG ANCA was detected in 88.7% and 11.1% had IgG+IgM and IgG1+IgG4 ANCA was detected in 50% patients. CONCLUSION: p-ANCA with anti-MPO is highly specific for MPA; both IIF and ELISA should be carried out for true positivity and to identify rare cases of dual specificities. Confocal laser scanning microscopy is useful in identifying ANCA patterns especially when ANA is also positive. ANCA testing with BVAS assessment will surely help in early diagnosis and estimating the severity of this life threatening disease.

Production & evaluation of a monoclonal antibody against human myeloperoxidase.

We describe the production of a mouse monoclonal antibody (H2E1) against human myeloperoxidase antigen. After production and characterisation, this antibody was compared with commercially available monoclonal antibodies, cytochemical myeloperoxidase and previously produced polyclonal antibody. Reaction with various cell lines proved that this monoclonal antibody was specific for myeloid lineage. This monoclonal showed positivity in 81.8 per cent of acute myeloid leukaemias whereas the polyclonal antibody was 100 per cent positive. We found that the polyclonal antibody was more sensitive as compared to the monoclonal. This is probably due to the lack of recognition of individual epitopes on the antigen. We recommend the use of antibodies which have different epitope recognition as most specific for myeloperoxidase.

Factores salivales de defensa no inmunoglobulínicos / Non-immunoglobulinic defense salivary factors

Este trabajo describe y analiza los principales factores antimicrobianos no inmunoglobulínicos de la saliva, su función biológica, interacciones y relación con diversas situaciones fisiológicas y patológicas que pueden presentarse en la cavidad bucal. Finalmente, se reseñan algunas aplicaciones clinicoterapéuticas de estos factores, destacando la necesidad de nuevas investigaciones destinadas a probar otros tratamientos que contribuyan a incrementar la efectividad de los sistemas innatos de defensa

Detección del antígeno viral de la rabia por el método avidina - biotina - peroxidasa / Detection of the rabies viral antigen by the avidin - biotin - peroxidase method

Antecedentes: La rabia humana y animal es frecuente en Colombia. Su diagnóstico requiere inmnofluorescencia directa (IFD) e inoculación intracerebral al ratón (IIC), condiciones que alcanzan pocos laboratorios. La histopatología muestra cuerpos de Negri entre el 40 por ciento al 87 por ciento de los casos. Nuestro objetivo es desarrollar y determinar la utilidad del método avidina-biotina-peroxidasa (ABP) en tejido fijado en formol e incluido en parafina. Materiales y métodos: Se estandarizó la técnica ABP usando muestras de encéfalo con rabia confirmada por IFD, IIC al ratón o histopatología. La técnica se pudo a prueba estudiando cortes de 100 encéfalos humanos o de animales sospechosos de tener rabia. Se escogieron encéfalos incluídos en parafina o mantenidos en congelación durante años, SNC fresco de ratón inoculado con virus fijo y cerebros normales o con otras encefalitis virales. Se determinó la sensibilidad, la especificidad y la reproducibilidad del método. Resultados: La técnica de ABP muestra sensibilidad mayor o igual a .80 y especificidad mayor o igual a .97. La concordancia con respecto a la IIC y a la IFD dio resultados entre substanciales y casi perfectos, según la escala de Landys y Koch, niveles altos que también se obtienen al evaluar la reproducibilidad intraobservador e interobservador. La técnica fue siempre mejor que la HE. Conclusiones: La técnica es útil, confiable, reproducible, fácil de realizar y funciona bien inclusive en encéfalos con pobre preservación tisular. Merece incluirse entre los procedimientos de rutina o alternativos en el dignóstico de la rabia humana y animal

Inmunodetección por peroxidasa de células de ganglio sensorial infectadas por virus de rabia / Immunodetection by peroxidasa of dorsal root ganglion by rabies virus infected cells

Con el objetivo de establecer una técnica de inmunocitoquímica por avidina-peroxidasa biotinilada para la detección de células infectas in vitro por el virus rábico, se usaron cultivos de ganglio de la raíz dorsal de ratón. En este trabajo se presentan los resultados del proceso de estandarización de la inmunoperoxidasa, en la que también se usa el conjugado antirrábico producido en el Instituto Nacional de Salud. Las diluciones de anticuerpo, que normalmente se usan en la inmunofluorescencia, muestran un excesivo ruido de fondo en la inmunoperoxidasa. Se discuten las posibles razones de este artefacto y se presenta un protocolo para la detección por inmunoperoxidasa de células de ganglio sensorial infectadas in vitro por virus de rabia

Anticuerpos anticitoplasma de neutrófilos (ANCA): su utilidad en el diagnóstico de las vasculitis autoinmunes / Anti-neutrophil cytoplasmic antibodies (ANCA): their usefulness in autoimmune vasculitis prognosis

Se determinó la asociación de ANCA con distintos tipos de entidades clínicas utilizando como técnica de tamizaje la inmunofluorescencia indirecta (IFI). Se determinó la especificidad antigénica utilizando enzimoinmunoanálisis (EIA). Se detectaron los isotipos de la inmunoglobulinas involucradas y su asociación con las manifestaciones clínicas. Para ello se examinaron los sueros de 102 pacientes con diagnóstico confirmado de vasculitis, de enfermedades del tejido conectivo, insuficiencia respiratoria y desórdenes renales. Se incorporó otra serie de 144 pacientes con distintas enfermedades que involucran o que pueden involucrar órganos y sistemas del mismo modo que las vasculitis inmunes. Del total de muestras, 29 fueron reactivadas por IFI y de éstas se estudiaron 16 por EIA, siendo 14 reactivas. Se detectó ANCA en 8/11 (73 por ciento) sueros de pacientes con granulomatosis de Wegener (GW); 7 presentaron cANCA (87,5 por ciento) y 1 pANCA. En Lupus Eritematoso Sistémico (LES), 4/32 (12,5 por ciento) fueron reactivos para ANCA (1 cANCA y 3 pANCA), y en Poliarteritis Nodosa (PAN) 6/8 (75 por ciento) fueron reactivos para pANCA. La imagen cANCA se asoció en forma significativa (p < 0,01) a GW respecto de las demás enfermedades y/o pacientes con manifestaciones clínicas relacionadas a vasculitis. Al agrupar los pacientes con LES, PAN, Hepatitis Autoinmune (3/8) e Insuficiencia Renal de causa desconocida (3/7), la imagen pANCA se asoció significativamente (p < 0,01) a estas enfermedades, en comparación al resto de los ANCA reactivos. El antígeno PR3 se asoció 1/1 con cANCA y 1/1 con pANCA, 12/16 MPO dieron imágenes pANCA y 2/12 imágenes pANCA no reaccionaron con ninguno de los antígenos estudiados. La asociación entre IgG ANCA con IgA e IgM Anca en el total de los pacientes fue no significativa (p > 0,01). En los tres casos clínicos reactivos para IgG e IgM se observó correlación entre su presencia y la afectación renal. El estudio sistemático de los ANCA y en particular de su especificidad antigénica, podrá sumar parámetros de evolución y pronóstico a los ya existentes y también mejorar el entendimiento de la fisiopatogenia de los fenómenos vasculíticos en diferentes enfermedades

Use of polyclonal anti-myeloperoxidase antibody in myeloid lineage determination.

This study reports the production of a rabbit polyclonal antibody to myeloperoxidase (MPO) and its use in ascertaining the myeloid lineage of blasts in leukaemia. Comparison of the immunocytochemical stain using the anti-MPO antibody with the routine cytochemical methodology showed that the former was more sensitive. In all subtypes of acute myeloid leukaemia (AML; 72 patients, M1-M6) greater number of MPO positive blast cells were observed by immunocytochemistry, the highest being in the promyelocytic leukaemia. It was also extremely specific for cells of the myeloid lineage as it did not react with blasts from acute lymphoblastic (50 patients) and megakaryoblastic leukaemias (1 patient). In addition, it proved most useful for the lineage determination of blasts from patients with undifferentiated acute leukaemias (AUL) and those with chronic myeloid leukaemia in blast crisis (CML-BC). Out of 8 patients of AULs, 6 were classified as acute myeloblastic leukaemia due to their reactivity to the anti-MPO antibody. Similarly, out of 12 patients of chronic myeloid leukaemia in blast crisis, blasts from 8 showed reactivity to this antibody and thus could be identified as belonging to the myeloid lineage and/or of the mixed blast crisis type.