Systemic lupus erythematosus: association with KIR and SLC11A1 polymorphisms, ethnic predisposition and influence in clinical manifestations at onset revealed by ancestry genetic markers in an urban Brazilian population.

Systemic lupus erythematosus (SLE) is an autoimmune disorder of the connective tissue with a wide and heterogeneous spectrum of manifestations, with renal and neurological involvement usually related to worse prognosis. SLE more frequently affects females of reproductive age, and a high prevalence and renal manifestation seem to be associated with non-European ethnicity. The present study aims to investigate candidate loci to SLE predisposition and evaluate the influence of ethnic ancestry in the disease risk and clinical phenotypic heterogeneity of lupus at onset. Samples represented by 111 patients and 345 controls, originated from the city of Belém, located in the Northern Region of Brazil, were investigated for polymorphisms in HLA-G, HLA-C, SLC11A1, MTHFR, CASP8 and 15 KIR genes, in addition to 89 Amerindian samples genotyped for SLC11A1. We also investigated 48 insertion/deletion ancestry markers to characterize individual African, European and Amerindian ancestry proportions in the samples. Predisposition to SLE was associated with GTGT deletion at the SLC11A1 3'UTR, presence of KIR2DS2 +/KIR2DS5 +/KIR3DS1 + profile, increased number of stimulatory KIR genes, and European and Amerindian ancestries. The ancestry analysis ruled out ethnic differences between controls and patients as the source of the observed associations. Moreover, the African ancestry was associated with renal manifestations.

Comparison between terminal slope rate constant and "slope/intercept" as measures of glomerular filtration rate using the single-compartment simplification.

Several whole-body variables are available for indexing glomerular filtration rate (GFR), including extracellular fluid volume (ECF volume, ECV), which has the advantage that GFR based only on the terminal exponential, alpha 2, of the chromium-51 ethylene diamine tetraacetic acid (51Cr-EDTA) plasma clearance curve is a measure already indexed for ECV. This slope-only method, however, is open to the theoretical criticism of a lack of robustness not present in the more conventional slope/intercept method, which generates GFR in absolute units before indexation to body surface area (BSA). To further evaluate the slope-only method, a large database of routine three-sample GFR measurements, based on 51Cr-EDTA, was analysed with the aim of comparing GFR/BSA with GFR/ECV, identifying the main sources of error in their respective measurements and explaining why they might be discrepant. The database was subdivided into 304 patients (group A) in whom the correlation coefficient of the fit to the three data points was greater than 0.99 and 31 patients in whom it was less than 0.99 (noisy data; group B). There was modest agreement between GFR/BSA and GFR/ECV in group A (r = 0.77; mean ratio 1.01 +/- 0.21). The difference between them correlated significantly with BSA (r = 0.52; P < 0.001), as would be expected given that small individuals have a relatively high BSA. Subdividing group A into four subgroups stratified according to BSA led to a slight improvement in the correlation between GFR/BSA and GFR/ECV. Because an error in alpha 2 is balanced by an opposing error in the intercept (which leads to a change in distribution volume in the opposite direction), and therefore impacts on both GFR/BSA and GFR/ECV, whereas an error in administered dose affects only GFR/BSA, it is possible to evaluate such errors by examining the respective relationships of GFR/BSA and GFR/ECV with indexed ECV. In group A, GFR/BSA correlated positively with ECV/1.73 m2, not surprisingly as ECV/BSA is effectively the ratio GFR/BSA to GFR/ECV, while GFR/ECV correlated negatively with ECV/1.73 m2. This implies that errors other than alpha 2, probably principally in administered dose, were as important as errors in alpha 2. This conclusion was supported by disappearance of the positive correlation between GFR/BSA and ECV/BSA in group B, brought about by the greater errors in alpha 2 in this group. This study suggests that, because of the effects of errors in the slope/intercept method that do not affect alpha 2, GFR based on slope only is at least as robust as that based on slope/intercept.

Indexed glomerular filtration rate as a function of age and body size.

The conventional way in which to scale or index a measurement of glomerular filtration rate (GFR) is to express it in relation to body surface area (BSA). However, BSA may not be appropriate for infants and children because, as individuals increase in size, their relative BSA decreases. Several other whole-body variables have been suggested as alternatives, including extracellular fluid volume (vECF). The purpose of the present study was to compare BSA and vECF as variables against which to index GFR, and in particular to look at this comparison in children versus adults. A total of 130 patients (age range 1-80 years; 40 patients <12 years) undergoing clinically indicated routine measurement of GFR using the bolus-injection single-compartment technique were included in the study. GFR was measured as the plasma clearance of [(51)Cr]EDTA as assessed from three peripheral venous blood samples taken between 2 and 4 h after injection of [(51)Cr]EDTA. Volume of distribution (V(d)) was obtained by extrapolation of the clearance curve to zero time. GFR was scaled to a BSA of 1.73 m(2). GFR and GFR/1.73 m(2) were corrected to account for the assumption of a single compartment. The rate constant of the exponential between 2 and 4 h was also corrected to give GFR/litre ECF. GFR and GFR/1.73 m(2) were both divided by GFR/litre ECF, to give vECF and vECF/1.73 m(2) respectively. Weight per unit BSA increases as a linear function of BSA. vECF is always less than V(d), on average by about 30%. vECF increased as an exponential function of BSA and as a linear function of body weight. vECF/70 kg body weight was higher in children (16. 2+/-3 litres) than adults (13.4+/-2.3 litres), but vECF/1.73 m(2) was lower in children (9.7+/-1.7 litres) compared with adults (12. 4+/-2 litres). vECV/1.73 m(2) increased as a function of both age and BSA, but vECF/kg decreased. GFR/12.5 litres vECF was higher than GFR/1.73 m(2) in children, but these values were similar in adults, with the ratio of these two forms of indexed GFR falling significantly with both age and BSA. Although this was not a normal population, but one with a wide range of renal function, GFR/vECF showed a strong inverse association with age, whereas for GFR/BSA the association was weak. In conclusion, these data provide further evidence that vECF is more valid physiologically for indexing GFR than is BSA, especially in children. Nevertheless, a GFR measurement in a child should ideally be expressed as a percentage of normal for that child's age. However, such normal values are not yet available.

Appropriate corrections to glomerular filtration rate and volume of distribution based on the bolus injection and single-compartment technique.

The bolus injection, single-compartment technique for measuring GFR overestimates the true value. Nevertheless, assuming that for a given indicator the area under the first exponential of the plasma clearance curve is constant from subject to subject, the observed (uncorrected) value can be corrected by multiplication with a 'sliding' factor, the value of which is a nonlinear function of GFR. Several second-order polynomials, based on pre-determined relationships between simultaneously determined two-compartment and one-compartment GFR, have been described for correcting GFR (GFR correction). It is, however, theoretically more rational to use a factor which depends on the rate constant, alpha2, of the terminal exponential of the clearance curve. We have therefore determined a set of linear equations from retrospectively analysed multiple-sample inulin, 99mTc-DTPA and 51Cr-EDTA clearance curves to enable correction of GFR using alpha2. A set of linear equations is also developed to correct the volume of distribution (Vd) of the indicator (close to extracellular fluid volume for these indicators), which is also overestimated by the one-compartment technique. At low levels of GFR, alpha2-corrected GFR is similar to uncorrected GFR for all three indicators. As GFR increases, however, uncorrected GFR progressively overestimates (alpha2-corrected GFR. The overestimation is greater for inulin than for 99mTc-DTPA or 51Cr-EDTA. In the one-compartment approximation, Vd is overestimated more than GFR, and again the greatest overestimation is seen with inulin. In a prospective study of 129 patients undergoing routine measurement of GFR with 51Cr-EDTA, alpha2 correction using a factor based on retrospective EDTA data gave values of GFR which were higher than values obtained from GFR correction using a previously published polynomial (also based on EDTA clearances) by 15% in children and 12.5% in adults when uncorrected GFR was 150 ml/min/1.73 m2. Moreover, the ratio of uncorrected GFR to GFR-corrected GFR was higher in children than adults. We conclude that alpha2 is a more rational variable with which to correct two-sample or three-sample GFR than GFR itself, that the correction formulae are not interchangeable between inulin on the one hand and EDTA and DTPA on the other, and that the relative magnitudes of the corrections given by alpha2 correction versus GFR correction are different for children and adults.

Inhaled platelet-activating factor causes pulmonary neutrophil sequestration in normal humans.

Inhaled platelet-activating factor (PAF) causes bronchoconstriction and transient peripheral neutropenia in humans. We studied eight normal subjects to investigate whether inhaled PAF caused pulmonary neutrophil sequestration. All subjects received autologous 99mTc-red cells as a blood pool marker, seven received 111In-neutrophils, and one received 111In-platelets. Six subjects inhaled 48 micrograms of PAF. There was immediate pulmonary sequestration of 111In-neutrophils, maximal (218% baseline) at 6 min (p less than 0.001), returning to normal by 3 h. There was no change in circulating platelet count or pulmonary 111In-platelet transit. Methacholine inhalation caused equivalent bronchoconstriction to PAF, but it had no effect on neutrophil count or pulmonary 111In-neutrophil activity. We have demonstrated pulmonary neutrophil, but not platelet, sequestration after PAF. This supports a role for PAF as an inflammatory mediator in humans. This may be a useful model for exploring pulmonary neutrophil kinetics and preinflammatory processes.

Quantification of right to left shunt at rest and during exercise in patients with pulmonary arteriovenous malformations.

BACKGROUND: Current treatment of patients with pulmonary arteriovenous malformations requires serial embolisations by means of steel coils or balloons. Measurement of right to left shunt is the most specific index of response to treatment. A new method of measuring shunt has been developed that is less invasive than traditional methods. METHODS: Right to left pulmonary shunt (expressed as percentage of cardiac output) was measured at rest in 19 patients with pulmonary arteriovenous malformations and six normal subjects by using intravenously injected albumin microspheres labelled with technetium-99m. The technique was compared with a simultaneous shunt measurement in subjects breathing 100% oxygen while they rested. The microsphere technique was adapted to measure the right to left shunt during exercise in 12 patients and five normal subjects with a new method of quantification. RESULTS: The mean (SD) shunt at rest as measured by the microsphere method was 23.2% (15.6%) in the patients and 2.7% (1.2%) in the normal subjects. When these values were compared with those of the 100% oxygen method the difference in mean values was 1% and the limits of agreement between the two methods -32% to +45%. The microsphere method is less invasive (arterial blood gas sampling is not required), quicker, and more comfortable for patients than the 100% oxygen method. In five of the normal subjects the mean (SD) 99mTc microsphere shunt increased from 2.9% (1.3%) at rest to 5.1% (2.9%) during exercise. In the 12 patients studied during exercise the shunt increased from 33.7% (12.7%) at rest to 41.7% (13.3%) during exercise in eight but decreased from 22.6% (2.4%) at rest to 17.6% (2.2%) during exercise in four. Arterial desaturation during exercise correlated with change in the size of the right to left shunt during exercise (r = +0.80). CONCLUSIONS: The microsphere method allows measurement of right to left shunt at rest and during exercise. Serial measurements at rest provide a simple, safe assessment of the physiological response to embolisation in patients with pulmonary arteriovenous malformations.

Renal haemodynamics of cyclosporin A nephrotoxicity in children with juvenile dermatomyositis.

1. Renal haemodynamics were monitored over an average period of 19 months in 17 children being treated with cyclosporin A. Sixteen had juvenile dermatomyositis and one had chronic polyneuropathy. The dose of cyclosporin A ranged from 2.3 to 8.3 mg day-1 kg-1 (median 4.1 mg day-1 kg-1). 2. Glomerular filtration rate (expressed in terms of extracellular fluid volume), renal blood flow (expressed as a fraction of cardiac output) and filtration fraction were measured by using 99mTc-labelled diethylenetriamine-penta-acetate. They were compared with the dosage and trough blood levels of cyclosporin A, and, in 15 patients receiving prednisolone in addition to cyclosporin A, with steroid dosage. 3. All 17 children had a renogram performed 6 months after starting cyclosporin A treatment. Nine of them also had a renogram before starting cyclosporin A treatment (baseline study), while 13, in addition to their renogram 6 months after starting cyclosporin A treatment, also had at least one further renogram. 4. Glomerular filtration rate/extracellular fluid volume fell slightly but significantly from 0.009 (SD 0.0013) before starting cyclosporin A treatment to 0.0085 (0.002) min-1 (P less than 0.01) 6 months after cyclosporin A treatment in the nine children who underwent a baseline study. This was accompanied by a significant (P less than 0.001) fall in filtration fraction from 0.108 (0.015) to 0.088 (0.014). However, renal blood flow/cardiac output showed no change. 5. In the 13 children studied beyond 6 months after starting cyclosporin A treatment, there was no further significant overall change in any renal haemodynamic variable.(ABSTRACT TRUNCATED AT 250 WORDS)

Inflammation: imaging with Tc-99m HMPAO-labeled leukocytes.

Leukocytes labeled with technetium-99m hexamethylpropyleneamine oxime (HMPAO) were used in 100 patients: 32 with suspected inflammatory bowel disease, 17 with fever of unknown origin, 21 with suspected abdominal sepsis, 20 with suspected bone sepsis, seven with bronchiectasis, and three with recent myocardial infarction. The distribution of activity in patients subsequently shown not to have inflammatory bowel disease was similar to that previously described for indium-111-labeled leukocytes. However, in this study, activity was also seen in the kidneys and bladder and occasionally the gallbladder on both early (1-3 hours) and late (24 hours) views, and in the colon in late views. Migration of Tc-99m-labeled granulocytes was seen in inflammatory disease as early as 30 minutes after injection, while normal bowel activity was not seen before 4 hours. The sensitivity of Tc99m-labeled leukocytes in the detection of inflammation was 100%, the specificity was 95%.

Noninvasive measurement of blood flow and extraction fraction.

We describe the theory of a technique for the noninvasive measurement of organ blood flow which is based on the principle of fractionation of cardiac output and is applicable with any recirculating gamma emitting tracer. The technique effectively determines the count rate that would be recorded over the organ if the tracer behaved like radiolabelled microspheres and was completely trapped in the organ's vascular bed on first pass. After correction for organ depth, the estimated first pass activity plateau, expressed as a fraction of the injected dose, is equal to the organ's fraction of the cardiac output (CO). By extending the theory, organ extraction fraction of extractable tracers or mean transit time of nonextractable tracers can be measured. Renal blood flow (RBF) to the normal left kidney in 18 subjects without evidence of renal disease was estimated by the technique to be 10.4% CO (S.D. 1.2), for the right kidney 9.0% CO(S.D. 1.1), and for both, 19.4% CO (S.D. 1.5). In a separate series of eight patients, RBF values were highly reproducible when obtained on successive days (standard deviation of change: 0.67% CO for the left kidney, 0.63% CO for the right and 0.78% CO for both). The extraction fraction of DTPA (filtration fraction) was 8.5% (1.7) in the left kidney and 9.7% (2.1) in the right kidney. This parameter was less reproducible than RBF, with standard deviations of the changes equal to 1.17% for the left kidney and 1.31% for the right. Stable, well-functioning, renal allografts in children had a mean blood flow of 20.6% CO (S.D. 3.7). Transplant blood flow in eight patients with rejection was less than 5.2% CO. In patients without splenomegaly, splenic blood flow, splenic extraction fraction of sulphur colloid and splenic red cell mean transit time were 4.3% CO (S.D. 0.9), 41% (S.D. 13.8), and 36.9 s (S.D. 4.6), respectively.

Clinical experience with 99mTc-hexamethylpropylene-amineoxime for labelling leucocytes and imaging inflammation.

Hexamethylpropylene-amineoxime (HMPAO) forms a lipid-soluble neutral complex with 99mTc which is rapidly incorporated into leucocytes in vitro. In six patients with suspected or known inflammatory disease, a "mixed" leucocyte suspension isolated from 85 ml blood anticoagulated with acid-citrate-dextrose was labelled by 99mTc-HMPAO with a mean efficiency of 47% (SE2%), of which 78% (3) was taken up by granulocytes. Activity eluted more rapidly from other cell types in vitro than from granulocytes, which remained firmly labelled. Mean initial biodistribution of the label and granulocyte recovery in blood of 32% (8) at 30-40 min showed that the granulocytes were not significantly activated during labelling. All six patients were positive for inflammatory disease, as early as 30 min in five patients and at 3 h in the sixth; they all remained positive at 20-24 h. Four patients also received 111In-labelled "pure" granulocytes. In terms of detail, the 99mTc images were comparable or superior to the 111In images.

Imaging of metastatic medullary carcinoma of the thyroid with 131I-meta-iodobenzyl guanidine.

The uptake of 131I-MIBG by medullary carcinoma of the thyroid was studied in six patients after total thyroidectomy in whom persistent raised plasma calcitonin levels were indicative of residual disease. The only patient in whom any activity in a possible metastasis was seen had by far the highest level of circulating calcitonin of the group and in contrast one patient with a presumptive metastatic deposit showed no uptake of MIBG into it. In conclusion the uptake of 131I-MIBG by medullary carcinoma of the thyroid is unreliable and unpredictable.