Assessment of changes in lactate concentration with intravascular microdialysis during high-risk cardiac surgery using the trend interchangeability method.

Background: Blood lactate is a strong predictor of mortality, and repeated blood lactate assays are recommended during surgery in high-risk patients. We hypothesized that the use of intravascular microdialysis incorporated in a central venous catheter would be interchangeable with the reference blood gas technique to monitor changes in blood lactate. Methods: Microdialysis and central venous blood lactate measurements were recorded simultaneously in high-risk cardiac surgical patients. The correlation between absolute values was determined by linear regression, and the Bland-Altman test for repeated measurements was used to compare bias, precision, and limits of agreement. Changes in lactate measurements were evaluated with a four-quadrant plot and trend interchangeability method (TIM). Results: In the 23 patients analysed, the central venous catheter was used as part of standard care, with no complications. The correlation coefficient for absolute values ( n =104) was 0.96 ( P <0.0001). The bias, precision, and limits of agreement were -0.19, 0.51, and -1.20 to 0.82 mmol litre -1 , respectively. The concordance rate for changes in blood lactate measurements ( n =80) was 94% with the four-quadrant plot. In contrast, the TIM showed that 23 (29) changes in lactate measurements were not interpretable, and among the remaining 57 (71) interpretable changes, 18 (32) were interchangeable, 8 (14) were in the grey zone, and 31 (54) were not interchangeable. Conclusions: Microdialysis with a central venous catheter appears to provide reliable absolute blood lactate values. Although changes in blood lactate measurements showed an excellent concordance rate, changes between the two methods were poorly interchangeable with the TIM. Clinical trial registration: NCT02296593.

Evaluation of cardiac troponin I stability in blood sample using the AccuTnI+3 assay.

OBJECTIVE: Troponins are considered as the biomarkers of choice to highlight cardiac injury in emergency departments, but are also valuable to detect cardiac injury in a non-emergency setting. In this latter case, transport of blood samples to laboratories often exceeds the manufacturer's recommendations (<2 h between vein puncture and analysis for the Beckman Coulter AccuTnI+3 assay). We aim to evaluate in vitro the stability of troponin Ic (cTnI) at two intervals (<2 h and at 4 h) over a wide range of concentrations using the Beckman Coulter AccuTnI+3 assay. PATIENTS AND METHODS: For each of the 95 patients included in this study, we analyzed the first blood sample with a time of transport <2 h, and the second sample after 4 h from vein puncture. We then calculated the correlation between the two periods of analysis and evaluated the bias by a Bland-Altman test. RESULTS: Taking into account of our analytical reproducibility, we did not observe any significant differences in cTnI values between <2 h and 4 h. CONCLUSIONS: The time between vein puncture and analysis of cTnI can be extended to 4 h.

Beware of suboptimal medical management of patients with atherosclerosis requiring revascularization.

AIM: The aim of this paper was to assess a daily-life evaluation of vascular risk factor control, pharmacological treatment and prognosis in patients with atherosclerosis referred for revascularization. METHODS: Prospective observational study conducted in a French University Hospital with collection of atherosclerosis referral site information, reported patient history, documented atherosclerosis sites seen on examination, biological data, and clinical outcomes. RESULTS: 956 patients (82.6% men, 64.5±10.1 years) were enrolled for supra-aortic vessel disease (SVD, 24.6%), coronary heart disease (CHD, 40.4%), peripheral artery occlusive disease (PAOD, 34.2%), and visceral artery disease (1.7%). Involvement of >2 vascular territories was documented in 85%. Vascular risk factor frequency results were: previous (65.7%) or current (10.6%) tobacco use, hypertension (64.3%), hyperlipidaemia (75.4%), diabetes (25.8%), overweight (43.8%), and obesity (25.2%). LDL-cholesterol was >100 mg/dL for 38.1%, most frequently seen in patients with PAOD referral (P<0.001) or history (P=0.002), and for 29.2% of the patients taking a statin. HbA1c levels were >6.5% for 53.8% of patients with diabetes. The triple combination of an antiplatelet agent, a statin, and a renin-angiotensin-system inhibitor was not prescribed often enough, especially for PAOD referrals (PAOD referrals, 45.1%; SVD referrals, 48.1%; CHD referrals, 65.9%). Independent risk factors for all-cause mortality were: a previous CHD or PAOD clinical event, body mass index <25 kg/m2, HbA1c >6.5%, and no aspirin treatment. CONCLUSION: Even at the time of revascularization, medical management of atherosclerosis was not optimal. The need for continuing education of physicians and patients remains essential.

Effectiveness of intensive insulin therapy by multiple daily injections and continuous subcutaneous infusion: a comparison study in type 2 diabetes with conventional insulin regimen failure.

OBJECTIVE: To compare the effectiveness of two intensified insulin regimens, i.e., pump delivery versus multiple daily injections in patients with type 2 diabetes not optimally controlled with conventional insulin therapy. RESEARCH DESIGN AND METHODS: Seventeen type 2 diabetes patients uncontrolled by two daily injections of regular plus NPH were randomly assigned in a cross-over fashion to either three daily injections of lispro plus NPH or pump device delivering lispro. HbA1c, 6 points capillary blood glucose, 24-hour continuous glucose monitoring system tracings and global satisfaction score were evaluated at the end of each 12-week treatment period. RESULTS: HbA1c decreased from 9.0+/-1.6% to 8.6+/-1.6% with multiple injections and 7.7+/-0.8% with pump device (p<0.03). Capillary blood glucose was lowered at all time-points with pump, but only at morning with multiple injections (p<0.01). Compared to conventional therapy, pump reduced hyperglycemic area under curve by 73% (p<0.01), but multiple injections by only 32% (p=0.08). Rate of hypoglycemia was not increased and patient's satisfaction was comparable with both intensive treatments. CONCLUSIONS: Pump therapy provides a better metabolic control than injection regimens, and seems to be safe and convenient in patients with type 2 diabetes who fail to respond to conventional insulin therapy.

[Multicenter evaluation of four homogenous LDL-cholesterol assays]. / Evaluation multicentrique de quatre méthodes de dosage direct du cholestérol-LDL.

International guidelines emphasize the importance of LDL cholesterol (LDL-C) assay in the care and follow-up of patients with cardiovascular risk. Most studies and common practice use Friedewald's formula for LDL-C calculation. The accuracy of the result depends closely on the precision of the input parameters (total cholesterol, triglycerides (TG) and HDL cholesterol), and discrepancies between calculated LDL-C and measurement by reference methods appear when TG exceed 4.5 mmol/L, or in the presence of abnormal lipoproteins. These restrictions and uncertainties in calculations have prompted the recent development of direct and homogeneous methods that fit all analyzers. A multicenter evaluation of four direct assays of LDL-C (Daiichi, Denka Seiken, Kyowa, Wako) was carried out on 45 serum samples (TG below 3.1 mmol/L) in eight laboratories using different analyzers. For three methods (Daiichi, Kyowa, Wako), the interlaboratory reproducibility was markedly improved relative to that of calculation. A strong correlation was found for all new methods when compared with a beta-quantification assay. Average bias in Denka Seiken assays was greater than Kyowa's and Daiichi's (although less dispersed for the latter) and for Wako all bias were positive. The relationship between bias variations and the lipid parameters of the samples was studied. Three methods, Daiichi, Kyowa and Wako, revealed a significant positive correlation between bias and serum VLDL-C/TG ratio, clearly indicating that cholesterol enrichment of VLDL was a source of variability in these assays. Specificity of the four methods was tested in situation of dyslipidemia by spiking isolated lipoproteins (chylomicrons, VLDL and HDL). This experiment revealed differences in behavior, most evidently upon addition of VLDL. No method was truly specific, but up to 8 mmol/L of TG the variations were acceptable. In the presence of type III hyperlipoproteinemia, however, only the Denka Seiken method was reliable. Linearity up to 20 mmol/L (Daiichi, Denka Seiken) or 14 mmol/L (Kyowa, Wako) of LDL-C allows these tests to be used in main routine cases. New direct assays are an obvious technological advance in terms of analytical performance and conveniency. Their use for the diagnosis and follow-up of hyperlipidemic patients offers an alternative that overcomes the limitations of the Friedewald calculation.

[Genetic polymorphism of the angiotensin I converting enzyme and morbi-mortality after myocardial infarction]. / Polymorphisme génétique de l'enzyme de conversion de l'angiotensine I et morbidité-mortalité après infarctus du myocarde.

Coronary atherothrombosis is a multifactorial disease dependent on environmental and genetic risk factors, not all of which have been identified. The renin-angiotensin system of the intermediary of the angiotensin I converting enzyme (ACE) plays a key role in cardiovascular physiopathology and contributes to phenomena of atherothrombosis. The concentrations of circulating ACE are partially determined genetically by the presence of polymorphism of insertion and deletion at the 16 intron of the gene. The implications of this polymorphism with respect to coronary disease have already been the object of many publications with contradictory results, usually in case-controlled studies. In this study, the authors sought to determine its influence on the morbi-mortality after myocardial infarction. A prospective cohort of 970 consecutive patients were followed-up for an average of 2.5 years. Genetic analysis was performed by a classical PCR protocol. The following independent predictive factors of mortality were the classical factors of age, hypercholesterolaemia and left ventricular dysfunction. However, the authors did not observe an effect of I/D polymorphism of the ACE gene on the occurrence of adverse cardiac events after myocardial infarction (death, infarction, revascularisation).

Prospective evaluation of the effect of an angiotensin I converting enzyme gene polymorphism on the long term risk of major adverse cardiac events after percutaneous coronary intervention.

OBJECTIVE: To evaluate prospectively the influence of an angiotensin I converting enzyme (ACE) gene polymorphism on long term clinical outcome of patients with established coronary artery disease treated by percutaneous coronary intervention. DESIGN AND SETTING: Prospective observational study in a university hospital. PATIENTS: Consecutive series of 1010 patients with symptomatic coronary artery disease who underwent successful coronary stent placement from November 1996 to April 1998. MAIN OUTCOME MEASURES: Long term clinical outcome was obtained and the rates of major adverse cardiac events (death, non-fatal acute myocardial infarction, unstable angina, and revascularisation) were compared according to the insertion/deletion (I/D) polymorphism of the ACE gene. RESULTS: Of the 1010 patients 29% had the DD genotype, 51% had the ID genotype, and 20% had the II genotype. All baseline clinical angiographic and procedural characteristics were identical in the three groups of patients. Event-free survival during the follow up period (median two years) was identical in patients with the II genotype compared to those with one or two D alleles. The predictors of long term survival were age, diabetes, ejection fraction, and extension of coronary artery disease. ACE genotype had no influence on the long term survival. Additional analyses assuming dominant and recessive effects of the D allele also failed to find any association; nor did the examination of low risk subgroups. CONCLUSIONS: The ACE I/D polymorphism does not influence the long term prognosis of patients with coronary disease treated by percutaneous coronary intervention, and screening patients for this gene polymorphism is not useful for secondary prevention strategies.

Relationship between polymorphisms in the renin-angiotensin system and nephropathy in type 2 diabetic patients.

BACKGROUND: Renin Angiotensin system is involved in renal function and its polymorphisms may influence diabetic nephropathy. ID ACE polymorphism modulates ACE level whereas M235T AGT polymorphism is involved in arterial hypertension. The A1166C AT1R polymorphism is involved in arterial hypertension and in diabetic retinopathy. METHODS: Two hundred thirty five type 2 diabetic patients were enrolled in this transversal study. Data were documented for clinical characteristics of the population, HbA(1c), urinary albumin excretion, presence of retinopathy or antihypertensive treatment. Polymorphisms were analyzed by PCR techniques. The patients were divided into 3 groups: group 1, without nephropathy (n=118), group 2, microalbuminuria (n=78), group 3, macroalbuminuria (n=39). RESULTS: Diabetes duration was longer (p<0.001), retinopathy (p<0.001) and antihypertensive treatment (p<0.02) were more frequent in group 3 compared to group 1 and 2. The I/D ACE and M235T AGT polymorphisms were not differently distributed between the three groups. In contrast, the CC genotype of the AT1R polymorphism was overrepresented in group 2 (p=0.021). The presence of the CC AT1R genotype considerably increased the incidence of albuminuria after 10 years of diabetes (AA vs CC p=0.01), particurlarly in men. No effect was seen with I/D ACE and M235T AGT polymorphisms. CONCLUSION: In conclusion, we observed an interaction of A1166C AT1R polymorphism with diabetes in men but not of I/D ACE and M235T AGT polymorphisms.

Child characteristics which impact accuracy of recall and suggestibility in preschoolers: is age the best predictor?

OBJECTIVE: The purpose of the present study was to determine whether individual difference factors of metamemory, intelligence, and temperament can improve the ability to predict accuracy of recall and suggestibility in preschoolers. METHOD: Fifty-six children ranging in age from 43 months to 83 months (M = 61, SD = 9) were recruited from 13 child care centers in a rural southeastern town. Children participated in a "Circus Day" event conducted by two female undergraduate psychology students dressed as clowns. Approximately 10 days (M = 10; SD = 2) after the event, children were interviewed regarding their experiences. RESULTS AND CONCLUSIONS: Bivariate correlations and multiple regression analyses were performed in order to determine which factors were related and unique contributors to accuracy of recall and suggestibility. Of principal importance is the finding that child characteristics such as metamemory ability, intellectual functioning, and temperament may indeed be helpful in determining a child's capacity to accurately recall information in an interview, although for the most part age is the best predictor. Findings also underscore the importance of considering a child's SES and race when planning and conducting interviews with young children. Possible explanations for these findings as well as implications for future research and clinical application are discussed.

Minimally invasive coronary surgery: surgical considerations and assessment of cardiac troponin I.

OBJECTIVE: Minimally invasive coronary artery bypass grafting (MICABG) using internal thoracic artery (ITA) without median sternotomy and cardiopulmonary bypass (CPB) become a viable option for the management of proximal left anterior descending artery (LAD) disease. Recent studies have demonstrated that cardiac troponine I (cTnI), a new highly specific diagnostic marker of cardiomyocyte damage, is a reliable marker of cardiac ischemia during heart operations under CPB. METHODS: Between February 1996 and April 1997, 14 patients (10 males, 4 females aged 41-68) underwent MICABG with single-vessel bypass grafting for LAD stenosis (n = 9) or occlusion (n = 5). Video-assisted surgery with left anterior mini-thoracotomy was performed in ten patients and vertical parasternal thoracotomy in the other four. cTnI was measured before LAD occlusion (T0), during anastomosis (T1) and 10 min (T2), 6 h (T3), 24 h (T4), 48 h (T5), 72 h (T6) after coronary reperfusion. Assay methods used a specific enzyme-linked immunosorbent autoanalyzer (Stratus) in peripheral venous blood. Control coronary angiography was performed in all patients. RESULTS: There were no operative complications, no reoperations for bleeding. cTnI concentrations were expressed in ng/ml +/- SD. Mean cTnI level was <3.85+/-1 ng/ml (range 0-32.8). Values were: T0 = 0, T1 = 0.5+/-0.1, T2 = 1.15+/-0.2, T3 = 2.16+/-0.6, T4 = 1.5+/-0.3, T5 = 0.6+/-0.02, T6 = 0.4+/-0.01. Angiography showed patent grafts in 12 patients. A 'no flow situation' was demonstrated in a cardiac symptom-free patient, with reestablishment of flow on repeat angiogram at 6 months. In the other case, early ITA graft occlusion in a patient with two-vessel disease was correlated with a higher cTnI concentration (17.8 ng/ml). Percutaneous angioplasty was performed on the right coronary artery, complicated with dissection and cardiac failure. This patient died 3 months after the MICABG despite ventricular assist device. CONCLUSION: cTnI did not increase during and after coronary artery occlusion and local immobilization of the heart. It can be used to evaluate postoperative myocardial damage on the beating heart using MICABG.

Safety of beating heart anastomosis during video-assisted coronary surgery attested by cardiac troponin I.

Our objective was to evaluate the safety of coronary anastomosis on the beating heart by measuring the release of cardiac troponin I during minimally invasive coronary artery bypass grafting (MICABG). Cardiac troponin I (cTnI) is a reliable marker of cardiac ischemia during heart operations under cardiopulmonary bypass (CPB). Ten patients (8 males and 2 females, aged 41-63) underwent MICABG with single vessel bypass grafting for left anterior descending coronary artery (LAD) stenosis (n = 7) or occlusion (n = 3). Video-assisted surgery with left anterior minithoracotomy was performed in all patients. Serial venous blood samples were collected for measurement of cTnI before LAD occlusion (T0), during anastomosis (T1) and 10 min (T2), 6 h (T3), 24 h (T4), 48 h (T5), and 72 h (T6) after coronary reperfusion. The assay method used a specific enzyme-linked immunosorbent Stratus autoanalyzer. Control coronary angiography was performed in all patients. There were no operative complications or reoperations for bleeding. The cTnI concentrations were expressed in ng/ml +/- SD. The mean cTnI level was less than 3.05 +/- 0.2 ng/ml (range 0-32.8). Values were T0 = 0, T1 = 0.4 +/- 0.03, T2 = 1.15 +/- 0.2, T3 = 2.16 +/- 0.6, T4 = 1.5 +/- 0.3, T5 = 0.6 +/- 0.02, and T6 = 0.4 +/- 0.01. Angiography showed patent grafts in 9 patients. In one case, early internal thoracic artery (ITA) graft occlusion in a patient with 2 vessel disease was correlated with a higher cTnI concentration (17.8 ng/ml). Percutaneous angioplasty was performed on the right coronary artery, complicated with dissection and cardiac failure. This patient died 3 months after the MICABG despite support from a ventricular assist device. In conclusion, collateral circulation developed in the setting of chronic coronary occlusion may be efficient for myocardial preservation during short periods such as coronary anastomosis. cTnI immunoassay confirmed the safety of coronary anastomosis on the beating heart during minimally invasive coronary operations.

Postprandial lipoprotein metabolism in normotriglyceridemic non-insulin-dependent diabetic patients: influence of apolipoprotein E polymorphism.

Non-insulin-dependent diabetes mellitus (NIDDM) is associated with postprandial lipoprotein clearance defects that are correlated with the fasting hypertriglyceridemia widely observed in NIDDM patients. The aim of this study was to determine if such postprandial disturbances are found in NIDDM patients strictly normotriglyceridemic in the fasting state, and if the apolipoprotein E (apo E) polymorphism influences postprandial metabolism of intestinally derived lipoproteins. The vitamin A-fat loading test was used in 18 normotriglyceridemic NIDDM patients and seven normotriglyceridemic obese controls, and postprandial triglyceride (TG) and retinyl palmitate (RP) concentrations were evaluated in total plasma, and in the chylomicron (Sf > 1,000) and nonchylomicron (Sf < 1,000) fractions isolated by ultracentrifugation. NIDDM patients exhibited an amplified response of both TG and RP as compared with obese controls in the three fractions. Incremental TG response to the oral fat load was strongly correlated with fasting TG level (r = .80, P < .0001) in the whole study population. Postprandial lipoprotein profiles were distinguished in NIDDM patients according to apo E phenotype: despite normal fasting TG levels in E3/3 (n = 6), E2/3 (n = 6), and E3/4 (n = 6), postprandial RP response was twofold to threefold higher in E2/3 and E3/4 patients than in the common E3/3 phenotype. Contrasting lower postprandial TG increment and lower fasting and postprandial high-density lipoprotein (HDL) and HDL3 cholesterol levels were observed in E3/4 versus E3/3 patients, possibly reflecting modifications in lipid content of the postprandial lipoproteins driven by a differential lipid transfer activity depending on apo E isoform. These data indicate an enhanced postprandial lipemia in normotriglyceridemic NIDDM patients, and demonstrate the influence of apo E polymorphism on their lipoprotein clearance. Postprandial alterations of lipoprotein remnants may thus accelerate atherogenesis even in normotriglyceridemic NIDDM patients.

[Physiological variations of LpA1, HDL2 and principal lipid markers in athletes]. / Variations physiologiques de la LpA1, des HDL2 et des principaux marqueurs lipidiques chez l'homme sportif.

An original approach of the influence of physical activities on lipid metabolism is presented in this work: the authors studied the physiological variations of the lipoparticle LpA1, high-density lipoprotein subfraction HDL2 and the most important lipid markers in serum of a presumably healthy population of 55 high-level sportsmen. They were 18-45 years old and practised endurance sports, whether individual (cycling, long-distance running), or collective activities (soccer, basketball). The authors observed an important increase of LpA1 (+20%, p < 0.001) and C-HDL2 (+23.3%, p < 0.01) after an intense physical activity (CK = 430 +/- 312 U/l); they also found a good correlation between LpA1 and HDL2 (r = 0.85, p < 0.0001) and between LpA1 and CK (r = 0.62, p < 0.001).

[Myoglobin and CPK-MM isoforms during the acute phase of myocardial infarction]. / Myoglobine et isoformes des CPK-MM à la phase aiguë de l'infarctus du myocarde.

The biological diagnosis is usually late with respect to clinical and electrocardiographic signs of acute myocardial infarction. Recent laboratory methods have stimulated renewed interest in very early marker of infarction: myoglobin and CPK isoforms. The authors undertook a prospective study of the diagnostic sensitivity of myoglobin (MG), CPK and CPK-MM isoforms in 30 consecutive patients undergoing intravenous thrombolytic therapy in the acute phase of myocardial infarction. Blood samples were obtained on admission and every 30 minutes for 1 h 30. The Mb contributed to diagnosis of infarction on admission in 89% of patients. This percentage fell to 44% for the ratio MM3/MM1 > 0.5 to 27% for the CPK and to 24% for the ratio MM3/MM1 > 1. The sensitivity of all tests increased in the later blood samples but it remained low in the CPK and MM3/MM1 > 1 criteria. The diagnostic sensitivity of the first sample was also better in patients admitted after the 3rd hour (Mb = 100%; MM3/MM1 > 0.5: 58%; CPK: 41%) compared with those admitted before the 3rd hour (Mb = 82%; MM3/MM1 > 0.5: 35%; CPK: 17%). These results show that the diagnostic sensitivity of biological markers of myocardial infarction is very different. The serum myoglobin is an earlier and more sensitive marker than the CPK or CPK-MM isoforms.

[Early clinical, electrocardiographic and biological criteria of reperfusion after intravenous fibrinolysis during the acute phase of myocardial infarction]. / Critères précoces, cliniques, électrocardiographiques, et biologiques de reperfusion après fibrinolyse intraveineuse à la phase aiguë de l'infarctus du myocarde.

Between May 1991 and February 1992, 31 consecutive patients were included in a prospective study, the aims of which were to determine the criteria of early coronary revascularisation after intravenous thrombolysis in the acute phase of myocardial infarction. The rise in serum myoglobin, the ST segment elevation, accelerated idioventricular rhythm and the evolution of chest pain were analysed. All patients underwent coronary angiography. Twenty-six were revascularized and 5 remained with coronary occlusion. Two types of serum myoglobin curves were demonstrated. Those with a sudden , decrease and a well defined peak in the first 4 hours were specific for revascularisation and easily identified (Group A: 16 patients). The graphs with a progressively rising slope to a peak after the 4th hour were observed in patients with coronary occlusion, but also in 10 patients with recanalized arteries (Group B). No significant difference was demonstrated with regards to the clinical and coronary angiographic parameters between patients in Group A and Group B. On the other hand, the time between the onset of chest pain and peak myoglobin was shorter in Group A (298 +/- 81 min) than in recanalised patients in Group B (380 +/- 54 min) (p < 0.05). The difference in the profile of the serum myoglobin could therefore reflect restoration of arterial flow in myocardial cells which had not suffered the same period of ischemia. ST segment elevation may increase, decrease of remain stable at 120 minutes in patients revascularised and those remaining occluded. In 9 patients, the ST elevation increased compared with the initial electrocardiogram .(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro effects of growth factors on rat germ cell RNA synthesis and their modulation by Sertoli cell-secreted proteins.

In vitro rat germ cell RNA synthesis is influenced by growth factors. Basic fibroblast growth factor (0.1 to 100 ng/ml) increases [3H]uridine incorporation in round spermatids (RS) but not in pachytene spermatocytes (PS); this effect is potentiated by insulin (10 micrograms/ml) and blocked in the presence of Sertoli cell-secreted proteins (SCSP). Somatomedin C (0.1 to 100 ng/ml) exhibits a similar effect when used alone without an influence by SCSP. Transforming growth factor beta (0.1 to 10 ng/ml) acts on both cell types, but SCSP amplify this effect only in PS. These data suggest that growth factors synthesized in situ may play a role in the germ cell development and that their effects are modulated by SCSP.

Validation of a new system for androgen binding protein measurement.

A new technique that permits measurement of Androgen-Binding Protein (ABP) is validated by reproducibility, linearity and correlation studies. Using this apparatus allowing Scatchard plot analysis, it is also possible to measure association and dissociation rate constants. In addition, it is a very useful tool for a rapid screening of ABP binding capacity during a chromatographic stepwise purification.

A minicolumn apparatus for androgen-binding protein measurement.

An easy, rapid, and sensitive assay that permits measurements of androgen-binding protein (ABP) in tissue as well as in spent media from Sertoli cells is described; this method involves the specific binding of labeled dihydrotestosterone (DHT) to ABP. The apparatus holds 36 minicolumns loaded with a DEAE Bio-Gel matrix. A peristaltic pump is used for the free fraction elution, taking into account the extremely rapid rate of dissociation of the ABP-DHT complexes. This technique, which allows Scatchard plot analysis, has been used to measure the rates of association (5.15 X 10(5)M-1 S-1 and dissociation (21.32 X 10(-4) S-1; t 1/2 = 5.5 min): the ratio of these rate constants is in perfect agreement with equilibrium dissociation constants determined by Scatchard plot analysis (KD = 4-4.5 nM). The intraassay and interassay coefficients of variation are 5 and 8%, respectively. A good correlation (r = 0.98) is obtained with the standard method of steady-state polyacrylamide gel electrophoresis below a value of 250 micrograms cytosolic proteins/gel. This apparatus, which allows either the measurement of ABP in 12 samples (in triplicates) at a saturating concentration or the analysis of two Scatchard plots (each of 6 points), is also very useful for a rapid localization of ABP during chromatographic purification.