Is there a role for urinary podocyte excretion assessment in lupus nephritis?

OBJECTIVE: To establish the occurrence and intensity of podocyturia and its relation to grade of disease activity, as defined by clinical and laboratory criteria. METHODS: Prospective, cross-sectional study involving 50 patients with lupus nephritis and 29 controls, which had podocyturia levels determined from random urine samples using an immunofluorescence technique. Disease activity was graded by BILAG (renal criteria) and an additional system used in the service (S2). RESULTS: Fifty patients with lupus nephritis (WHO classes III, IV and V), with a median age of 37 years, were evaluated. Of these, 86.5% were female, and 52% were BILAG A. Podocyturia quantification in the lupus nephritis and control groups differed significantly (p = 0.009). This score was higher in relation to classes III, IV and V. The correlation with C3 consumption was stronger (p = 0.011) than with C4. The highest levels were found in the most active groups (A and B of BILAG and S2). Lower podocyturia correlated with a lower dose of prednisone. There was no association with the intensity of proteinuria, hematuria or pyuria, serum creatinine levels, among others. CONCLUSIONS: Podocyturia assessment, which was performed by immunofluorescence in this study, can be used as an indicator of disease activity with the advantage of being a urinary biomarker. The levels proved to be higher in patients with lupus nephritis than in the controls and were particularly higher in class IV.

It is time to review concepts on renal involvement in leprosy: pre- and post-treatment evaluation of 189 patients.

BACKGROUND: Functional and morphological renal lesions have been widely described in leprosy for decades. Nevertheless few studies have assessed renal function pre- and during treatment after the advent of multidrug therapy (MDT). METHODS: This is a prospective study involving 189 consecutive patients, with all forms of leprosy (Ridley-Jopling scale). Laboratory (serum urea and creatinine, estimated GFR, urinalysis, microalbuminuria, urinary RBP) and clinical features of renal disease were evaluated previously and after onset (3 and 8 months later) of MDT. RESULTS: One hundred and eighty-nine patients (M 1.8: F 1; mean age 44 ± 16 years) were included just after diagnosis of leprosy and before the introduction of MDT. Mean time until manifestation of symptoms and/or signs of leprosy was 29 ± 56 months (25 days-480 months). Microhematuria and microalbuminuria were detected in 7.5% and 9.6% of the cases, respectively. Elevated serum creatinine was detected in 34% pre-MDT; this was statistically more frequent in males, hypertensive and frequent users of non-steroidal anti-inflammatory drugs (NSAID), as well as in patients with erythema nodosum lepromatosum, 45.5% by the time of diagnosis, 18% after 3 months and 9% after 8 months of MDT (p = 0.039). CONCLUSIONS: Our results suggest that functional renal lesions in leprosy are currently mild and predominantly of glomerular origin, in opposition to the severe involvement described in the past. This improved outcome of renal disease secondary to leprosy is possibly due to the advent of MDT and effective treatment of episodes of reaction, leading to shorter periods of active infectious disease.

Estimation of glomerular filtration rate from serum creatinine and cystatin C in octogenarians and nonagenarians.

BACKGROUND: Equations to estimate GFR have not been well validated in the elderly and may misclassify persons with chronic kidney disease (CKD). We measured GFR and compared the performance of the Modification of Diet in Renal Disease (MDRD), the Chronic Kidney Disease-Epidemiology Collaboration (CKD-Epi) and the Berlin Initiative Study (BIS) equations based on creatinine and/or cystatin C in octogenarians and nonagenarians. METHODS: Using cross-sectional analysis we assessed 95 very elderly persons (mean 85 years) living in the community. GFR was measured by iohexol (mGFR) and compared with estimates using six equations: MDRD, CKD-Epi_creatinine, CKD-Epi_cystatin, CKD-Epi_creatinine-cystatin, BIS_creatinine and BIS_creatinine-cystatin. RESULTS: Mean mGFR was 55 (range,19-86) ml/min/1.73 m(2). Bias was smaller with the CKD-Epi_creatinine-cystatin and the CKD-Epi_creatinine equations (-4.0 and 1.7 ml/min/1.73 m(2)). Accuracy (percentage of estimates within 30% of mGFR) was greater with the CKD-Epi_creatinine-cystatin, BIS_creatinine-cystatin and BIS_creatinine equations (85%, 83% and 80%, respectively). Among the creatinine-based equations, the BIS_creatinine had the greatest accuracy at mGFR < 60 ml/min/1.73 m(2) and the CKD-Epi_creatinine was superior at higher GFRs (79% and 90%, respectively). The CKD-Epi_creatinine-cystatin, BIS_creatinine-cystatin and CKD-Epi_cystatin equations yielded the greatest areas under the receiver operating characteristic curve at GFR threshold = 60 ml/min/1.73 m 2 (0.88, 0.88 and 0.87, respectively). In participants classified based on the BIS_creatinine, CKD-Epi_cystatin, or BIS_creatinine-cystatin equations, the CKD-Epi_creatinine-cystatin equation tended to improve CKD classification (net reclassification index: 12.7%, p = 0.18; 6.7%, p = 0.38; and 15.9%; p = 0.08, respectively). CONCLUSIONS: GFR-estimating equations CKD-Epi_creatinine-cystatin and BIS_creatinine-cystatin showed better accuracy than other equations using creatinine or cystatin C alone in very elderly persons. The CKD-Epi_creatinine-cystatin equation appears to be advantageous in CKD classification. If cystatin C is not available, both the BIS_cr equation and the CKD-Epi_cr equation could be used, although at mGFR < 60 ml/min/1.73 m(2), the BIS_cr equation seems to be the best alternative.

Autoantibodies against phospholipase A2 receptor in Brazilian patients with glomerular diseases.

BACKGROUND: Recently, great progress has been made in understanding the pathogenesis of membranous nephropathy (MN) with the discovery of autoantibodies (Abs) to M-type phospholipase A2 receptor (PLA2R) in serum and in immunocomplexes deposited in glomerulus in most adult patients with primary MN. OBJECTIVE: To evaluate the diagnostic performance of anti-PLA2R in Brazilian patients with MN, as well as to verify the possible association of anti-PLA2R serum levels with disease activity. METHODS: 117 patients with glomerular diseases confirmed by renal biopsy underwent routinely clinical and laboratory evaluation (serum creatinine and albumin, 24-h proteinuria, urinalysis, tests for etiological investigation) and determination of serum anti-PLA2R by ELISA. RESULTS: 67.5% of the patients had MN, 9.4% focal segmental glomerulosclerosis, 7.7% lupus nephritis class V and 15.4%, other proteinuric glomerular diseases. The mean level of glomerular filtration rate (estimated by the CKD-EPI formula) was 79.43 mL/min (12.00-151.20 mL/min), 24 h proteinuria of 2.89 g (0-14.90 g), serum albumin of 3.79 g/dL (1.20-4.80 g/dL). Anti-PLA2R was detected in 27 patients, all with active MN, being 26 primary and 1 secondary MN. Sensitivity and specificity rates for the test were 60.5-94.7%, and positive (PPV) and negative (NPV) predictive values were 92.9 and 67.9%, respectively. CONCLUSIONS: Anti-PLA2R showed high specificity and PPV for the diagnosis of primary MN in Brazilian patients. There was a strong correlation between disease activity and positive anti-PLA2R. This biomarker represents an important diagnostic tool for primary MN and may contribute to the monitoring of disease activity in such patients.

Pregnancy in renal transplant patients: Renal function markers and maternal-fetal outcomes.

OBJECTIVES: We aimed to evaluate laboratory markers in women who got pregnant after renal transplantation. STUDY DESIGN: Cross-sectional prospective study. MAIN OUTCOME MEASURES: Renal function parameters and maternal and fetal data were assessed in renal transplant recipients. RESULTS: Forty-three women who got pregnant after renal transplantation (mean age, 28.5 years; mean gestational age, 35.6 weeks) were included. Most patients (53.5%) received a renal transplant from a deceased donor. Podocyturia was not significantly correlated with other renal function markers. Mean period from transplantation to pregnancy was approximately 5 years; this period was not associated with obstetric complications or changes in renal markers. A gradual increase was observed in the following parameters during pregnancy and puerperium: serum creatinine levels (P < 0.001), proteinuria (P < 0.001), urinary protein/creatinine ratio (P < 0.001), and albumin/creatinine ratio (P < 0.001). The sensitivity and specificity of protein/creatinine ratio in predicting preeclampsia were high (96.0% and 94.0%, respectively). Elevated serum creatinine levels, urinary albumin/creatinine ratio, and retinol-binding protein levels in the third trimester were associated with prematurity (P < 0.001). Preeclampsia was the main cause of renal function decline at the end of pregnancy (65.0% of cases). Approximately four (9.5%) pregnant women presented with premature rupture of membranes and 18 (42.0%) with a urinary tract infection. CONCLUSIONS: Proteinuria, urinary protein/creatinine ratio, and retinol-binding protein levels were elevated in patients with preeclampsia. Using these markers to assess renal function during pregnancy may be clinically useful for detecting and monitoring renal injury in renal transplant recipients.

GFR estimated from cystatin C versus creatinine in children born small for gestational age.

BACKGROUND: Low birth weight caused by intrauterine growth restriction may be a risk factor for renal impairment in the adult life. STUDY DESIGN: A cross-sectional study. SETTING & PARTICIPANTS: 71 children aged 8 to 13 years living in the community of São Paulo, Brazil, were included in the study. Gestational age was within the normal range. PREDICTORS: Birth weight (range, 2,052 to 3,560 g) divided into quartiles: 2,500 g or less; 2,501 to 2,740 g; 2,741 to 3,000 g; and greater than 3,000 g. Birth weight ascertained by birth records in 43 and by recall in 28 participants. OUTCOMES & MEASUREMENTS: Cystatin C, creatinine, and glomerular filtration rate (GFR) estimated by equations using cystatin C (eGFR(cys)) or creatinine (eGFR(cr)). RESULTS: Overall, mean serum creatinine level was 0.8 +/- 0.01 (SE) mg/dL (range, 0.7 to 1.1 mg/dL); mean plasma cystatin C level was 0.9 +/- 0.02 mg/L (range, 0.5 to 1.6 mg/L), and eGFR(cr) and eGFR(cys) were 102.4 +/- 2.16 (range, 66 to 140) and 91.8 +/- 2.46 mL/min/1.73 m(2) (range, 49 to 139 mL/min/1.73 m(2)), respectively. No differences were found for serum creatinine or eGFR(cr) values among the birth-weight quartiles. There was a significant linear trend of increasing cystatin C levels (decreasing eGFR(cys)) in the lower birth-weight quartile groups (P = 0.002 and P = 0.02, respectively). Systolic blood pressure correlated with plasma cystatin C level (r = 0.31; P = 0.008) and eGFR(cys) (r = -0.26; P = 0.028). Covariance analysis adjusting for age, sex, body mass index for age compared with standards of the National Center for Health Statistics and expressed as a z score, and systolic blood pressure showed that cystatin C values remained greater in the lowest than highest birth-weight quartile (1.01 +/- 0.05 versus 0.83 +/- 0.05 mg/L; P = 0.02). LIMITATIONS: Ascertainment of birth weight by recall in some participants. Lack of measurement of microalbuminuria, absence of direct GFR measurement, and small sample size. CONCLUSIONS: Lower birth weight is associated with higher levels of cystatin C but not creatinine in 8-13 yr. old children born full-term.

Are urinary levels of free light chains of immunoglobulins useful markers for differentiating between systemic lupus erythematosus and infection?

INTRODUCTION: Active systemic lupus erythematosus (SLE) and infection are often hard to distinguish. We evaluated the urinary levels of free light chains of immunoglobulins (urFLCIg) as a possible laboratory marker to differentiate those conditions. METHODS: We evaluated 43 patients with lupus nephritis (16-63 years old), with or without concurrent infection (12 with infection), 14 with infectious disease without SLE and 20 with idiopathic Fanconi's syndrome. The Systemic Lupus Erythematosus Disease Activity Index was utilized to establish activity of disease. Levels of urFLCIg kappa and lambda were determined by an immunoenzymometric assay developed in our institution. In order to evaluate proximal tubular dysfunction which could be responsible for increased levels of urFLCIg, we determined the low-molecular-weight protein urRBP. RESULTS: Levels of urFLCIg in healthy volunteers (median kappa 1.57 mg/l; lambda 0.96 mg/l), inactive SLE (5.36; 4.93) and active SLE (11.82; 23.59) were significantly different; urFLCIg levels or urFLCIg/urRBP ratios could not separate patients with infection from those with SLE. CONCLUSION: Our data show that convoluted proximal tubular dysfunction was not responsible for the increase in urFLCIg levels. UrFLCIg determination was useful in the detection of SLE activity, but was unable to distinguish activity from infection in this condition.

[Unexpected high values of TSH: the presence of high molecular weight forms (macro TSH) must be investigated]. / Valores inesperadamente elevados de TSH: a presença de formas de alto peso molecular ("macro TSH") deve ser investigada.

The laboratory methods usually employed for the measurement of serum TSH present sensitivity and specificity levels, both analytical and clinical, are highly satisfactory. Additionally, the methodologies are quite robust, so that false-positive and false-negative results are rare and unexpected. In this paper we describe two individuals quoted as euthyroid clinically, with no reference to autoimmune diseases, and no reference to the use of exogenous TSH, that presented with normal to extremely high serum TSH levels, depending on the method employed for analysis. In the three tested methods, serial dilution showed that the real TSH levels were between 250 and 300 mUI/L. In both cases the increment in TSH levels were due to the presence of TSH-binding proteins, forming high molecular weight complexes ("macro TSH"), well characterized by gel filtration chromatography on Superdex S-200 column. In one of the patients the binding protein was characterized as being IgG by protein-G binding study. In the other case, protein-G binding as well as anti-IgM binding failed to characterize the protein. These two cases call attention to the importance of the clinical-laboratory correlation and suggest the need that the presence of "macro TSH" must be investigated in patients with unexpectedly high TSH values.

Urinary abnormalities and renal function in pregnant women with chronic hypertension. / Avaliação de alterações urinárias e função renal em gestantes com hipertensão arterial crônica.

INTRODUCTION: Renal involvement in pregnant women with chronic hypertension is not widely known. OBJECTIVES: 1- To describe the epidemiological profile of pregnant women with chronic hypertension; 2- To evaluate urinary abnormalities (by urinalysis), renal function (serum creatinine and cystatin C, and estimated glomerular filtration rate (eGFR); 3- To evaluate the pregnancy outcome in chronic hypertension. METHODS: 103 pregnant women with chronic hypertension (blood pressure over 140/90 mmHg, detected previously to pregnancy or until the 20th week) were submitted to clinical and laboratorial evaluation. RESULTS: Pregnant women were 21-45 (mean: 34) years-old. Protein/creatinine ratio in random urine was elevated in 5.2% (0.0-6.4g/g), serum creatinine in 19.6% and cystatin C in 14.7% of them. It was observed that characteristics of pregnant patients and their newborns (vs. frequencies of the cases with CKD-EPI cystatin C < 60 ml/min/1.73 m2) were: 20.5% (33.3%) of preterm birth < 37 weeks, 17.5% (22.2%) of birth weight < 2500g and 17.5% (22.2%) of small for gestational age; superimposed preeclampsia-eclampsia occurred in 24.7% (22.2%) of the cases. CONCLUSIONS: Renal abnormalities were detected by proteinuria, determinations of serum creatinine and cystatin C in 5.2, 19.6 and 14.7% of the cases. The results suggest that the formulas CKD-EPI and MDRD can have applicability in assessing renal function in pregnant women. It was also shown a high frequency of preterm birth or with < 2500g at birth or small for gestational age, as well as of superimposed preeclampsia-eclampsia (24.7%) in pregnant women with chronic hypertension.

Iohexol clearance for determination of glomerular filtration rate in rats induced to acute renal failure.

INTRODUCTION: The glomerular filtration rate (GFR) is considered an especially important tool for the measurement of renal function. Inulin clearance (InCl) is the classic reference method for this purpose, although it is associated with a number of disadvantages; thus, other markers have been proposed, including iohexol. Determination of iohexol clearance (IoCl) has been established for clinical use; however, its application as a GFR marker in experimental rat models has not been reported. OBJECTIVES: This study aims to standardize a methodology for the measurement of iohexol clearance and to evaluate its applicability as a marker of GFR in rats with induced toxic acute renal failure (ARF), using InCl as the gold standard. MATERIALS AND METHODS: Twenty-six Wistar male rats (200-300 g) were divided into the following two groups: a control group (n=7) and an ARF group (n=19). ARF was induced by the subcutaneous administration of cisplatin (5 mg/kg); IoCl and InCl were determined simultaneously, and plasma creatinine (pCreat) dosage was measured colorimetrically. RESULTS: The pCreat, InCl and IoCl levels were consistent with the expected values for the renal function ranges of the evaluated animals, and the IoCl and InCl levels were significantly correlated (r=0.792). An inverse moderate linear correlation between the IoCl and pCreat measurements (r=-0.587) and between the InCl and pCreat measurements (r=-0.722) were observed. CONCLUSION: These results confirm a correlation between IoCl and the gold standard of GFR, InCl measurement. IoCl offers a relevant advantage over InCl because determination of the former allows the animal to live after the procedure.

Pre- and post-transplant anti-myosin and anti-heat shock protein antibodies and cardiac transplant outcome.

BACKGROUND: The purpose this study was to investigate the relationship of anti-myosin and anti-heat shock protein immunoglobulin G (IgG) serum antibodies to the original heart disease of cardiac transplant recipients, and also to rejection and patient survival after cardiac transplantation. METHODS: Anti-myosin and anti-heat shock protein (anti-hsp) IgG antibodies were evaluated in pre-transplant sera from 41 adult cardiac allograft recipients and in sequential post-transplant serum samples from 11 recipients, collected at the time of routine endomyocardial biopsies during the first 6 months after transplantation. In addition, the levels of these antibodies were determined from the sera of 28 healthy blood donors. RESULTS: Higher anti-myosin antibody levels were observed in pre-transplant sera than in sera from normal controls. Moreover, patients with chronic Chagas heart disease showed higher anti-myosin levels than patients with ischemic heart disease, and also higher levels, although not statistically significant, than patients with dilated cardiomyopathy. Higher anti-hsp levels were also observed in patients compared with healthy controls, but no significant differences were detected among the different types of heart diseases. Higher pre-transplant anti-myosin, but not anti-hsp, levels were associated with lower 2-year post-transplant survival. In the post-transplant period, higher anti-myosin IgG levels were detected in sera collected during acute rejection than in sera collected during the rejection-free period, whereas anti-hsp IgG levels showed no difference between these periods. CONCLUSIONS: The present findings are of interest for post-transplant management and, in addition, suggest a pathogenic role for anti-myosin antibodies in cardiac transplant rejection, as has been proposed in experimental models of cardiac transplantation.

[Parathyroid hormone values obtained with immunometric assays depend on the amino-terminal antibody specificity]. / Valores de paratormônio obtidos com ensaios imunométricos dependem da especificidade do anticorpo amino terminal empregado.

Introduction of 2nd generation immunometric assays for the measurement of serum parathyroid hormone (PTH), turned them more available, simple and rapid. These methods, based on double identification of the PTH molecule, supposedly measure the intact, bioactive molecule, with the sequence 1-84. Recent works showed that they also measure forms with amino-terminal deletions, like the 7-84 form, which are not able to activate the traditional PTH receptor (PTH1R). Thus, an important practical aspect is the definition of the PTH forms measured by the immunometric assays, a fact that depends on the specificity of the antibodies employed. In this report we compare the results obtained with an in-house immunofluorometric assay that presents a cross-reactivity of 50% with the 7-84 PTH sequence, and two commercial 2nd generation assays, that react 100%. In a first study, 135 samples were measured using our assay and an electrochemiluminescent assay, resulting in a correlation coefficient of 0.961 (P<0.0001) and medians of 35.0 and 51.0 ng/L (P<0.0001). In a second study, 252 samples were analyzed using our assay and an immunochemiluminometric assay, resulting in a correlation of 0.883 (P<0.0001) and medians of 36.0 and 45.5 ng/L (P<0.0001). In both studies results obtained with the in-house assay were significantly lower, as expected by the specificity of the anti-amino-terminal antibody employed. Our data support the need of a precise description of the specificity of the amino-terminal antibodies employed in 2nd generation PTH assays in order to better compare results and define normal ranges.

Serum Beta 2-microglobulin/cystatin C index: a useful biomarker in lupus nephritis?

BACKGROUND: Systemic lupus erythematosus (SLE) is a chronic multisystem autoimmune disease with frequent flares. Our aim was to evaluate the beta 2-microglobulin/cystatin C (ß2M/CysC) index versus other markers as a predictor factor for assessment of SLE reactivation. METHODS: We prospectively analyzed 42 patients with lupus nephritis. Disease activity was classified using SLEDAI-2K and BILAG. Routine renal function and laboratory markers of SLE activity were performed, as well as serum ß2M (Sß2M)/serum CysC (SCysC) and Sß2M/serum creatinine (SCreat) indexes determinations. RESULTS: The 42 enrolled patients had a mean age of 37.7 ± 13.1 years, 88% were female and 67% Caucasians; mean estimated glomerular filtration rate was 61.9 ± 20.0 ml/min/1.73 m(2). There was a strong correlation between SCreat versus SCysC (r = 0.887), SCreat versus Sß2M (r = 0.865), and SCysC versus Sß2M (r = 0.880). Multivariate analysis showed that the Sß2M/SCreat index is a prognostic factor predicting active lupus nephritis. CONCLUSION: As SCysC is a good marker of renal function, it would be expected that the Sß2M/SCysC index could be a better indicator of renal activity than Sß2M/SCreat, but in the present study it did not add relevant clinical information in the assessment of renal activity in SLE.

Race adjustment for estimating glomerular filtration rate is not always necessary.

BACKGROUND: Estimated glomerular filtration rate (eGFR) is very important in clinical practice, although it is not adequately tested in different populations. We aimed at establishing the best eGFR formulas for a Brazilian population with emphasis on the need for race correction. METHODS: We evaluated 202 individuals with chronic kidney disease (CKD) and 42 without previously known renal lesions that were additionally screened by urinalysis. Serum creatinine and plasma clearance of iohexol were measured in all cases. GFR was estimated by the Mayo Clinic, abbreviated Modification of Diet in Renal Disease (MDRD) and Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) formulas, and creatinine clearance was estimated by the Cockcroft-Gault (CG) formula. Plasma clearance of iohexol was used as the gold standard for GFR determination and for the development of a Brazilian formula (BreGFR). RESULTS: Measured and estimated GFR were compared in 244 individuals, 57% female, with a mean age of 41 years (range 18-82). Estimates of intraclass correlation coefficients among the plasma clearance of iohexol and eGFR formulas were all significant (p < 0.001) and corresponded to the following scores: CG 0.730; obesity-adjusted CG 0.789; Mayo Clinic 0.804; MDRD 0.848; MDRD1 (without race adjustment) 0.846; CKD-EPI 0.869; CKD-EPI1 (without race adjustment) 0.876, and BreGFR 0.844. CONCLUSIONS: All cited eGFR formulas showed a good correlation with the plasma clearance of iohexol in the healthy and diseased conditions. The formulas that best detected reduced eGFR were the BreGFR, CKD-EPI, and CKD-EPI1 formulas. Notably, the race correction included in the MDRD and CKD-EPI formulas was not necessary for this population, as it did not contribute to more accurate results.

Avaliação de alterações urinárias e função renal em gestantes com hipertensão arterial crônica / Urinary abnormalities and renal function in pregnant women with chronic hypertension

Resumo Introdução: O acometimento renal em gestantes portadoras de hipertensão arterial crônica (HAC) não é amplamente conhecido. Objetivos: 1- Descrever o perfil epidemiológico de pacientes com HAC; 2- Avaliar a ocorrência de alterações urinárias e de função renal (por meio de determinação sérica de creatinina, cistatina C e ritmo de filtração glomerular estimada - RFGe); 3- Avaliar o desfecho das gestações em HAC. Métodos: Foram submetidas a avaliações clínicas e laboratoriais 103 gestantes com HAC (pressão arterial acima de 140/90 mmHg, identificada previamente à gestação ou até a 20ª semana). Resultados: As gestantes tinham 21-45 (média: 34) anos; 12,6% eram primigestas, 64,1% tiveram múltiplas gestações. A relação proteinúria/creatininúria em amostra isolada estava alterada em 5,2% casos (0-6,44 g/g), creatinina sérica estava elevada em 19,6% e cistatina C em 14,7%. Na avaliação das características da gestação em pacientes com HAC e seus recém-nascidos (RN) (vs. frequências nos casos com CKD-EPI cistatina C < 60 ml/min/1,73 m2), observou-se: 20,5% (33,3%) de nascidos pré-termo < 37 sem, 17,5% (22,2%) de RN com peso < 2500 g e 17,5% (22,2%) de RN pequeno para a idade gestacional (PIG); sobreposição de DHEG ocorreu em 24,7% (22,2%) dos casos. Conclusão: Alterações renais foram identificadas por proteinúria, creatinina e cistatina C séricas em 5,2%, 19,6 e 14,7% das gestantes. Os resultados sugerem que as fórmulas do CKD-EPI e MDRD também podem ter aplicabilidade nessa avaliação em gestantes. Detectou-se alta frequência de RN pré-termo ou com menos de 2500 g ao nascer ou PIG, assim como de sobreposição de DHEG (24,7%) em gestantes com HAC.

Abstract Introduction: Renal involvement in pregnant women with chronic hypertension is not widely known. Objectives: 1- To describe the epidemiological profile of pregnant women with chronic hypertension; 2- To evaluate urinary abnormalities (by urinalysis), renal function (serum creatinine and cystatin C, and estimated glomerular filtration rate (eGFR); 3- To evaluate the pregnancy outcome in chronic hypertension. Methods: 103 pregnant women with chronic hypertension (blood pressure over 140/90 mmHg, detected previously to pregnancy or until the 20th week) were submitted to clinical and laboratorial evaluation. Results: Pregnant women were 21-45 (mean: 34) years-old. Protein/creatinine ratio in random urine was elevated in 5.2% (0.0-6.4g/g), serum creatinine in 19.6% and cystatin C in 14.7% of them. It was observed that characteristics of pregnant patients and their newborns (vs. frequencies of the cases with CKD-EPI cystatin C < 60 ml/min/1.73 m2) were: 20.5% (33.3%) of preterm birth < 37 weeks, 17.5% (22.2%) of birth weight < 2500g and 17.5% (22.2%) of small for gestational age; superimposed preeclampsia-eclampsia occurred in 24.7% (22.2%) of the cases. Conclusions: Renal abnormalities were detected by proteinuria, determinations of serum creatinine and cystatin C in 5.2, 19.6 and 14.7% of the cases. The results suggest that the formulas CKD-EPI and MDRD can have applicability in assessing renal function in pregnant women. It was also shown a high frequency of preterm birth or with < 2500g at birth or small for gestational age, as well as of superimposed preeclampsia-eclampsia (24.7%) in pregnant women with chronic hypertension.

Structural and mutational analyses of protein-protein interactions between transthyretin and retinol-binding protein.

Transthyretin is a tetrameric binding protein involved in the transport of thyroid hormones and in the cotransport of retinol by forming a complex in plasma with retinol-binding protein. In the present study, we report the crystal structure of a macromolecular complex, in which human transthyretin, human holo-retinol-binding protein and a murine anti-retinol-binding protein Fab are assembled according to a 1 : 2 : 2 stoichiometry. The main interactions, both polar and apolar, between retinol-binding protein and transthyretin involve the retinol hydroxyl group and a limited number of solvent exposed residues. The relevance of transthyretin residues in complex formation with retinol-binding protein has been examined by mutational analysis, and the structural consequences of some transthyretin point mutations affecting protein-protein recognition have been investigated. Despite a few exceptions, in general, the substitution of a hydrophilic for a hydrophobic side chain in contact regions results in a decrease or even a loss of binding affinity, thus revealing the importance of interfacial hydrophobic interactions and a high degree of complementarity between retinol-binding protein and transthyretin. The effect is particularly evident when the mutation affects an interacting residue present in two distinct subunits of transthyretin participating simultaneously in two interactions with a retinol-binding protein molecule. This is the case of the amyloidogenic I84S replacement, which abolishes the interaction with retinol-binding protein and is associated with an altered retinol-binding protein plasma transport in carriers of this mutation. Remarkably, some of the residues in mutated human transthyretin that weaken or abolish the interaction with retinol-binding protein are present in piscine transthyretin, consistent with the lack of interaction between retinol-binding protein and transthyretin in fish.

Urinary retinol-binding protein as a prognostic marker in glomerulopathies.

Tubulointerstitial involvement seems to have a decisive influence on the progression of glomerular diseases. We have prospectively evaluated the levels of urinary retinol-binding protein (urRBP), a marker of proximal tubular dysfunction, in patients with different glomerulopathies (GPs) and correlated these levels with disease progression. By studying 238 patients with GPs, we found that urRBP tend to be lower in minimal change disease, glomerular hematuria and poststreptococcal glomerulonephritis as compared to focal segmental glomerulosclerosis, membranous nephropathy and membranoproliferative glomerulonephritis. By following 149 patients for up to 10 years, we have concluded that high levels of urRBP can identify patients who will progress with loss of renal function (defined as doubling of serum creatinine level) and that a urRBP level >1 mg/l was an efficient and independent indicator of poor prognosis as shown by multivariate analysis. This prediction was possible at a time when serum creatinine and creatinine clearance were still in the normal range. Our data suggest that this laboratory test adds important clinical information to the follow-up of GPs.

Valores inesperadamente elevados de TSH: a presença de formas de alto peso molecular ("macro TSH") deve ser investigada / Unexpected high values of TSH: the presence of high molecular weight forms (macro TSH) must be investigated

As metodologias empregadas para a medida de TSH sérico apresentam níveis de especificidade e sensibilidade, tanto diagnósticas como analíticas, bastante elevadas. Adicionalmente, são metodologias bastante robustas, de maneira que resultados falso-positivos ou falso-negativos são raros e inesperados. Descrevemos neste trabalho dois casos de indivíduos descritos como eutiróideos, sem antecedentes de doenças autoimunes e sem referência ao uso de TSH exógeno, que apresentavam níveis de TSH de normais a muito elevados, dependendo da metodologia empregada. Em três métodos testados para a medida de TSH a diluição seriada das amostras mostrou que os níveis reais situavam-se entre 250 e 300 mUI/L. Nos dois casos, o aumento de TSH foi ocasionado pela presença de proteínas ligadoras de TSH no soro, formando formas de alto peso molecular ("macro TSH"), demonstradas por cromatografia de gel filtração em coluna de Superdex S-200. Em uma das pacientes a proteína ligadora foi caracterizada como IgG através de cromatografia em proteína G sepharose, na outra, a ligação em proteína G e em coluna de sepharose acoplada a monoclonal anti-IgM não conseguiu caracterizar a proteína ligadora sérica. Estes casos salientam a importância da correlação clínico-laboratorial e sugerem a necessidade de se pesquisar a presença de macro TSH em pacientes com níveis de TSH anormalmente elevados.

Desenvolvimento de ensaio imunofluorométrico para a medida da globulina ligadora de tiroxina (thyroxine-binding globulin, TBG) e sua aplicaçäo em casos de deficiência desta proteína / Development of an immunofluorometric assay for thyroxine-binding globulin (TBG) and its application in cases of protein deficiency

A globulina ligadora de tiroxina (thyroxine-binding globulin, TBG) é a principal transportadora de hormônios tiroidianos no soro. Variaçöes na concentraçäo sérica de TBG determinam variaçöes proporcionais nas concentraçöes séricas totais de T4 e T3, sem implicar alteraçöes de funçäo, desde que a fraçäo livre permaneça normal. Várias condiçöes clínicas comuns levam a alteraçöes significativas nos níveis de TBG, sendo as variaçöes mais importantes devidas a defeitos genéticos. Como a TBG é codificada por gene localizado no cromossomo X, os defeitos se manifestam mais facilmente no sexo masculino. Descrevemos o desenvolvimento de ensaio imunofluorométrico para a medida de TBG com base em anticorpos monoclonais, sendo um desenvolvido em nosso laboratório e outro comercial. O método apresenta sensibilidade de 0,8mg/l e coeficientes de variaçäo intra e interensaio inferiores a 10 por cento. O estudo comparativo com método comercial mostrou alta correlaçäo (r = 0,93; n = 48), sendo os valores normais obtidos de 10mg/l a 29mg/l. Estudamos também 20 indivíduos portadores de deficiência congênita de TBG, 19 homens e uma mulher, que apresentavam valores normais de TSH e baixos de T4 total; em todos eles os níveis de TBG foram indetectáveis. Já os níveis de T4 livre medidos por método indireto em 16 desses indivíduos mostraram-se elevados em todos, ao passo que, quando medidos por método direto pós-diálise nos quatro restantes, mostraram-se normais. Nossos resultados reforçam a necessidade prática da disponibilidade de ensaio para a medida de TBG para esclarecimento e definiçäo diagnóstica de alguns casos especiais, principalmente quando o ensaio de T4 livre direto, pós-diálise, näo é disponível