Successful treatment for a case of near-fatal secondary adrenal insufficiency / Tratamiento exitoso de una caso de insuficiencia adrenal secundaria casi fatal

A thirty-six-year old female with shock was found to be unconsciousness a few days after developing a respiratory infection. Her past medical history included autoimmune hypothyroidism. Her state of shock was not controlled by massive fluid resuscitation with a vasopressor and antibiotics. However, an infusion of 250 mg methylprednisolone dramatically improved her shock state. Further examination indicated secondary acute adrenal insufficiency. Adrenal insufficiency may complicate other endocrine disorders. Accordingly, a physician should consider hypoadrenocorticism, when patients are in a state ofrefractory shock in spite ofmassive infusion with a vasopressor, especially in patients with other endocrine disorders.

Una mujer de treinta y seis años en shock fue hallada inconsciente unos dias después de desarrollar una infección respiratoria. Los antecedentes en su historia clinica incluian hipotiroidismo autoinmune. Su estado de shock no fue controlado por la reanimación con liquidos masiva con un vasopresor y antibióticos. Sin embargo, una infusion de 250 mg metilprednisolona habia mejorado considerablemente su estado de shock. Un examen mas detenido indicó insuficiencia adrenal aguda secundaria. La insuficiencia adrenal puede complicar otros trastornos endocrinos. En consecuencia, un médico debe considerar la posibilidad de hipoadrenocorticismo, cuando los pacientes se encuentran en estado de shock refractario a pesar de una infusion masiva con un vasopresor, especialmente en el caso pacientes con otros trastornos endocrinos.

Determination of renal function in long-term heart transplant patients by measurement of urinary retinol-binding protein levels

Significant improvements have been noted in heart transplantation with the advent of cyclosporine. However, cyclosporine use is associated with significant side effects, such as chronic renal failure. We were interested in evaluating the incidence of long-term renal dysfunction in heart transplant recipients. Fifty-three heart transplant recipients were enrolled in the study. Forty-three patients completed the entire evaluation and follow-up. Glomerular (serum creatinine, creatinine clearance measured, and creatinine clearance calculated) and tubular functions (urinary retinol-binding protein, uRBP) were re-analyzed after 18 months. At the enrollment time, the prevalence of renal failure ranged from 37.7 to 54 percent according to criteria used to define it (serum creatinine > or = 1.5 mg/dL and creatinine clearance <60 mL/min). Mean serum creatinine was 1.61 ± 1.31 mg/dL (range 0.7 to 9.8 mg/dL) and calculated and measured creatinine clearances were 67.7 ± 25.9 and 61.18 ± 25.04 mL min-1 (1.73 m²)-1, respectively. Sixteen of the 43 patients who completed the follow-up (37.2 percent) had tubular dysfunction detected by increased levels of uRBP (median 1.06, 0.412-6.396 mg/dL). Eleven of the 16 patients (68.7 percent) with elevated uRBP had poorer renal function after 18 months of follow-up, compared with only eight of the 27 patients (29.6 percent) with normal uRBP (RR = 3.47, P = 0.0095). Interestingly, cyclosporine trough levels were not different between patients with or without tubular and glomerular dysfunction. Renal function impairment is common after heart transplantation. Tubular dysfunction, assessed by uRBP, correlates with a worsening of glomerular filtration and can be a useful tool for early detection of renal dysfunction.

Serum leptin concentration during puberty in healthy nonobese adolescents

Data obtained during the past five years have indicated that there are important age- and gender-based differences in the regulation and action of leptin in humans. To study the physiological changes of leptin during puberty in both sexes, and its relationship with body composition and sexual maturation, we measured leptin concentrations in 175 healthy adolescents (80 girls, 95 boys, 10-18 years of age), representing all pubertal stages. We excluded individuals with a body mass index (BMI) below the 5thor above the 95th percentile relative to age. Serum concentrations of leptin were determined by a monoclonal antibody-based immunofluorimetric assay, developed in our laboratory. Body composition was determined by dual-energy X-ray absorptiometry. Pubertal stage was assigned by physical examination, according to Tanner criteria for breast development in females and genital development in males. Leptin concentration in girls (N = 80) presented a positive linear correlation with age (r = 0.35, P = 0.0012), BMI (r = 0.65, P < 0.0001) and percentfat mass (r = 0.76, P < 0.0001). In boys (N = 95) there was a positive correlation with BMI (r = 0.49, P < 0.0001) and percentfat mass (r = 0.85, P < 0.0001), but a significant negative linear correlation with Tanner stage (r = -0.45, P < 0.0001) and age (r = -0.40, P < 0.0001). The regression equation revealed that percentfat mass and BMI are the best parameters to be used to estimate leptin levels in both sexes. Thus, the normal reference ranges for circulating leptin during adolescence should be constructed according to BMI or percentfat mass to assure a correct evaluation

A specific and highly sensitive time-resolved fluoroimmunoassay for human proinsulin

We describe a time-resolved fluoroimmunoassay specific for human proinsulin using a combination of two high-affinity monoclonal antibodies, one against insulin and the other specific for intact proinsulin and for split 65-66 and des 64-65 proinsulin forms. The assay employs only 200 micro liters of serum, with a detection limit of 0.1 pmol/l. The intra-assay variation coefficient was less than 3 percent between 3 and 1000 pmol/l. There was 0 percent cross-reaction with insulin, C-peptide, split 32-33 and des 31-32 proinsulin. Serum concentration of proinsulin was analyzed in 50 subjects during an oral glucose tolerance test (10 non-obese controls, 10 obese controls, 10 subjects with impaired glucose tolerance, 10 patients with type II diabetes meIlitus (DM) and fasting blood glucose (FBG) <140 mg/dl, and 10 patients with type II DM and FBG >150 mg/dl). Mean fasting serum proinsulin levels measured by this assay in non-obese controls (0.84 +/-0.90 pmol/l; 0.1-2.4 pmol/l) were lower than the results reported by her investigators. There was an increase of proinsulin related to obesity and increased glucose levels, suggesting that proinsulin levels increase with insulin resistance.

Monoclonal antibodies specific for the free alpha subunit of glycoprotein hormones and their use in the development of a sensitive immunofluorometric assay

Glicoprotein hormone free alpha subunit has been used as a marker for some pituitary tumors and to study the reactivity of glycoprotein hormone-producing cells under different circunstances. We describe a highly sensitive ans specific immunofluorometric assau for the measurement of serum free alpha subunit levels. The assay is based on a monoclonal antibody, specific for free alpha subunit, bound to microtiter plates. As tracer antibody we employed an europium-labelled free/complexed alpha subunit specific monoclonal antibody. Using overnight incubation and 50µl samples, the least detectable dose was of the order of 4 ng/1. Cross-reactivity with LH, TSH, FSH, and hCG was 6.5, 1.2, 4.3 and 1.1 percent, repectively. Normal adult males showed values ranging from 120 to 790ng/l, not different from normal adult premenopausal females (88 to 604 ng/l). In post-menopausal females, serum concentrations were significantly highler, ranging from 341 to 407 ng/l. In 56 patients with untreated pituitary tumors (18 "non-secreting", 25 GH-producing and 13 prolactin-producing tumors), 10 showed high values, 3 of them from the first group, 3 from the second and 4 from the third. We conclude that this highly sensitive assay can be a valualbe tool for the diagnosis and follow-up of selected patients with pituatary tumors and in other circumstances in which the glycoprotein hormone-producing cells of the pituitary require evaluation

Comparison of the determination of insulin by a monoclonal antibody-based immunofluorometric assay and by radioimmunoassay

This paper describes an immunofluorometric assay (IFMA) for insulin and compares it with the classical radioimmunoassay (RIA). Monoclonal antibodies against insulin were produced and used to develop the IFMA. One, immobilized on microtiter plates, was used for capture, the other, labelled with Europium, was used as tracer antibody. The IFMA presentes sensitivity to an amount of insulin of 3 pmol/1 and acceptable valueus for intra- and interassay error. The IFMA presented superimposable curves for human insulin, Arg65/Gly66-split proinsulin and des-Lys64, Arg65, and no cross-reactivity with human proinsulin, Arg32/Glu33 -split and des-Arg31, Arg32. The RIA showed 100 percent cross-reactivity with human proinsulin, 90 pecent with des-Arg31, Arg32 and 170 percent with des-Lys64, Arg65. The assay were used to measure insulin in 300 serum samples from 50 subjects submitted to an oral glucose tolerance test (OGTT). Twenty were normal, 10 had impaired glucose tolerance and 20 non-insulin-dependent diabetes mellitus. The mean value (ñ SEM) obtained bu IFMA was 166.7 ñ 12.1 pmol/1 and the mean value obtained by RIA was 339.6 ñ 18.6, with a correlacion of r = 0.80 (P0.01). Comparison of basal insulin levels of the different groups of individuals using IFMA or RIA led to the same conclusions. The area under curve showed statistically significant differences only for the comparison between normal lean subjects and individuals with impaired glucose tolerance, when measured by RIA...(au)

Development and clinical application of an immunoflurometric assay for intact parathyroid hormone

Parathyroid hormone (PTH) is a linear peptide of 84 amino acids that is found in serum mainly in the form of carboxyl-terminal fragments. The biological activity of PTH depends on the presence of the amino-terminal portion and in circulation is limited to the intact molecule. We describe an immunmofluorometric assay for the measurement of PTH-(1-84) based on a chicken egg yolk-derived amino-terminal antibody bound to microtiter plates by an anti-chicken Ig monoclonal antibody. As tracer antibody we employed a Europium-labelled carboxyl-terminal specific monoclonal antibody produced from a mouse immunized with hPTH-(53-84)-BSA conjugate. The assayincluded an initial overnight incubation of the sample and the solid phase-bound amino-terminal antibody, followed by washing and addition of the tracer antibody, and an additional two hours of incubation prior to fluorescence reading. The least-detectable dose was in the order of 2.5 pg/.ml and preliminary studies in 40 normal adults showed values in the range of 4 to 70 pg/ml; fo 12 patients with surgery-proven primary hyperparathyroidism values ranged from 109 to 743 pg/ml and for 34 patients with humoral hypercalcemia of malignancy from 2.5 to 66 pg/ml. We conclude that this assay, with its increased sensitivity and specificity, will be a valuable tool in the study of PTH secretion in normal and pathological situations

ß2 microglobulina (ß2M) sérica na monitorizaçäo do transplante renal / Serum ß2 microglobulin (ß2M) following renal transplantation

Apesar da melhora na sobrevida do enxerto em pacientes transplantados renais, ocorrida nos últimos dez anos, a rejeiçäo continua sendo uma causa importante de perda de enxerto. Vários testes laboratoriais têm sido estudados na tentativa de se identificar um método näo invasivo que possibilite o diagnóstico precoce de rejeiçäo em pacientes transplantados renais. OBJETIVO. Avaliar a utilidade da monitorizaçäo da ß2 microglobulina sérica no período inicial pós-transplante. Métodos. foram estudados em 20 receptores de transplante renal (10 doadores vivos relacionados e 10 doadores cadáveres), o comportamento diário dos níveis séricos da ß2M e correlacionados com a sua evoluçäo clínica e laboratorial. RESULTADOS. Pacientes que apresentaram boa funçäo renal no pós-operatório imediato e näo mostraram rejeiçäo aguda, precocemente, evoluíram com queda dos níveis de ß2M que se estabilizaram em níveis de 3,7 mg/L no 4§ dia pós-transplante. A sensibilidade de ß2M para o diagnóstico de rejeiçäo aguda foi muito boa (87,5 por cento), mas sua especificidade foi baixa (46 por cento). Nos oito pacientes que näo apresentaram boa funçäo renal, inicialmente, a monitorizaçäo dos níveis de ß2M mostrou-se capaz de diferenciar pacientes com necrose tubular aguda (NTA) sem complicaçöes, de portadores de NTA evoluindo com rejeiçäo ou nefrotoxicidade por CSA. Conclusäo. A monitorizaçäo dos níveis séricos de ß2M näo acrescenta benefício nítido para o diagnóstico de rejeiçäo aguda em pacientes com boa funçäo renal inicial. Contudo, em pacientes evoluindo para NTA, esta monitorizaçäo mostrou-se útil para identificar episódios de rejeiçäo aguda e nefrotoxicidade por CSA

Specific and nonspecific aspects of humoral immune response in leprosy

1. We have studied some generic and specific aspects of the humoral immune response in 96 patients with leprosy (29 paucibacillary and 67 multibacillary individuals). We determined serum immunoglobulins (IgM, IgG and IgA), CH50, C1q, C3 and C4, circulating immune complexes (CIC), C-reactive protein (CRP), rheumatoid factor (RF) and antinuclear antibodies. No specific pattern of general humoral immune changes could be observed. 2. The specific immune response was studied by the detection of specific IgM anti-M. leprae antibodies. An immunoradiometric assay (IRMA) and an ELISA were compared for clinical effectiveness. IRMA showed greater sensitivity for the serodiagnosis of leprosy as compared to ELISA (88.1 percent vs 58.2 percent for multibacillary patients and 20.7 percent vs 10.3 percent for paucibacillary leprosy patients). Specificity was 96 percent for IRMA and 97 percent for ELISA. 3. Our results indicate that nonspecific changes in the humoral immune response are of little value in assessing leprosy patients and that immune assays for the detection of specific anti-M. leprae antibodies may be of value in the diagnosis, study and follow-up of these patients

Clinical utility of a 22-kDa growth hormone-specific assay

Human growth hormone (hGH) circulates in different molecular forms, with the 22-kDa monomer being the predominant one and the 20-k-Da variant corresponding to 5 to 15% of the serum hGH on a weight basis. Using monoclonal antibodies with different specificities we developed two immunoenzymometric assays, one with 22 + 20 k-Da specificity and the other specific only for the 22-kDa form. Both assays used microtiter plates as solid phase and streptavidin-peroxidase for color development; intra-assay CV was less than 10% in the range of 1 to 100 mlU/l for the 22 + 20 kDa assay and in the range of 3 to 100 for the 22-kDa assay, with an inter-assay CV of less than 14% for both assays, sensitivity was 0.2 mlU/l for the 22 + 20 kDa assay and 0.5 mlU/l for the 22-kDa assay. The two assays were compared by measuring 200 serum samples with detectable hGH levels by both assays. Higher values were obtained with the 22 + 20 kDa assay (62.1 ñ 59.2 ñ 6.1 mlU/l, mean ñ SD) with a correlation coefficient (r) of 0.99. In no clinical condition (28 patients with growth retardation and 14 acromegalics) did the two assays give discrepant values. We conclude that there was no practical advantage in using an assay with specificity restricted to the 22-kDa form for measuring hGH in clinical serum samples

Monoclonal antibody-based immunoenzymometric assay for serum human growth hormone

A monolconal antibody-based immunoenzymometric assay (IEMA) for the measurement of human serum growth hormone is described. Two high-affinity and complementary monoclonal antibodies were selected from a panel of 9 obtained upon fusion of SP/O myeloma cells with spleen cells from a Balb/c mouse immunized against human growth hormone of pituitary origin. One monoclonal antibody was immobilized by attaching it to the walls of microtiter wells and the second was biotinylated. The reaction was quantitated by the addition of streptavidin-peroxidase. The sensitivity of the assay was 0.2 mIU/1 and the intra-and interassay coefficients of variation for 4.6 to 46 mIU/I wee < 8.3 and 17.3%, respectively. Cross-reaction with human placental lactogen, human prolactin and rat growth hormone was < 0.1% (w/w). Comparison of results obtained for 180 routine serum assays by radioimmunoassay and the assay described here had a correlation coefficient of 0.94 with a mean value of 16.3 ñ 1.3 (X ñ SEM) and 13.3 ñ 1.2 mIU/!, with the IEMA providing values 18% lower than the RIA. The discrepancy emphasizes the necessity of redefining normal ranges before immunometric assays, like the one described, can be used routinely