Comparative study of three vaginal markers of the premature rupture of membranes. Insulin like growth factor binding protein 1 diamine-oxidase pH.

OBJECTIVE: To assess the diagnostic value of three vaginal markers-insulin-like growth factor binding protein 1 (= IGFBP1), diamine-oxidase (= DAO) and pH-for diagnosis of the premature rupture of membranes. STUDY: One hundred pregnant women participated in the study. They were divided into three groups: group A (34 cases with intact membranes), group B (35 cases with total rupture of the membranes), group C (31 cases of suspected rupture of the membranes). Each patient underwent three successive tests for each of the three markers. The test order was allocated at random. For pH the reaction is colorimetric, for DAO the reaction is radio-enzymatic and for IGFBP1 the reaction is immuno-chromatographic. All three reactions are qualitative in nature. The parameters studied were conventional statistical parameters (sensitivity = SN, specificity = SP, positive predictive value = PPV and negative predictive value = NPV). RESULTS: The analysis of the statistics gave the following results in percentages for SN, SP, PPV and NPV respectively; pH: 90.7%, 77.2%, 75%, 91.7%. DAO: 83.7%, 100%, 100%, 89%. IGFBP1: 95.3%, 98.2%, 97.6%, 96.5%. CONCLUSION: The determination of variations in pH is not satisfactory. IGFBP1 is at least better than DAO with, additionally, advantages of rapidity and simplicity.

Progress report of an external quality assessment scheme for cyclosporine assay.

Beginning in 1987, an external quality assessment (EQA) scheme for cyclosporine (CsA) assay was organized by our Institute and supported by National Research Council (CNR); in 1991, the CNR EQA joined the French CsA interlaboratory program organized by Service de Radiopharmacie et Radioanalyse, University of Lyon. During the last 2 years (1993 and 1994 cycles), > 170 laboratories (from seven European countries) participated in this survey, assaying 44 control samples prepared from pooled blood of heart- and renal-transplant patients; normal pools added with known amounts of CsA were also distributed. During the whole EQA period, a trend toward the use of specific and nonisotopic techniques has been observed. In 1994, 91% of the collected results have been produced by specific methods [high-pressure liquid chromatography (HPLC) or specific immunoassays developed to assay the native molecule of CsA without its metabolites];in the same cycle, the fully automated techniques [TDx, fluorescence polarization immunoassay (FPIA), and enzyme-multiplied immunoassay (EMIT)] accounted for 73% of total results. CsA concentration measured by monoclonal specific immunoassays [(TDx FPIA, radioimmunoassay (RIA) Cyclo-Trac, and EMIT] was well correlated with those from HPLC (r = 0.99-1.00); results from EMIT were very close to those from HPLC, whereas results from RIA Cyclo-Trac and TDx were slightly overestimated (10-20%). Nonspecific methods (TDx polyclonal and nonspecific RIA Cyclo-Trac) measured CsA concentrations 3-4 times higher than those found by HPLC. The cross-reactivity of CsA metabolites in specific immunoassays was estimated from data of patients' samples and spiked samples; an interference of 6% in RIA Cyclo-Trac, 7% in EMIT, and 26% in TDx FPIA was found. The precision coefficient of variation (CV% between laboratory and between assay) of the methods observed in the 1994 EQA cycle was 9.5 for TDx polyclonal FPIA, 10.4 for TDx monoclonal FPIA, 10.5 for EMIT, 10.6 for RIA specific Cyclo-Trac, 14.2 for RIA nonspecific Cyclo-Trac, and 15.2 for HPLC.

Comparison between chronic converting enzyme inhibition and AT1 blockade in mRen2 transgenic rats.

We compared the consequences of chronic angiotensin-converting enzyme (ACE) inhibition with quinapril and of specific AT1 blockade with losartan in a renin-dependent model of hypertension, the (mRen2)27 transgenic rats (TG). Animals were orally treated with 10 mg/kg/24 h of either quinapril (TGQ, n = 13) or losartan (TGL, n = 12) from age 4 to age 9 weeks. Indirect systolic blood pressure (SBP), and sodium and water balances were measured for 3 consecutive weeks. Nine-week-old rats were instrumented to record aortic BP in the conscious state. In addition, they received an infusion of glucose and saline to increase their diuresis and thus allow accurate assessment of their renal excretion during short time periods. These rats were studied for three one-h periods: (a) baseline, (b) after the administration of a bradykinin (BK) antagonist, and (c) after a cross-treatment; i.e., TGQ rats receiving losartan (10 mg/kg intravenously, i.v.) and TGL rats receiving quinapril (10 mg/kg i.v.). TGL rats differed from TGQ rats by an unconsistently lower indirect SBP associated with significantly lower urinary volume and sodium excretion, whereas the sodium balance did not differ between the two groups. In conditions of fixed sodium intake the aortic BP of TGQ rats was still nonsignificantly different from that of TGL rats, and TGQ rats also exhibited two-fold higher natriuresis. The BK antagonist had no effect in either group, whereas losartan decreased the BP of TGQ rats. We conclude that in TG rats ACE inhibition is associated with an increased natriuresis as compared with specific AT1 blockade, an effect that is independent of the sodium intake. Because a BK antagonist had no effect, such a difference might be due to an antinatriuretic effect of AT2 receptors in chronic conditions.

Performance of immunoassays for ca 19-9, ca 15-3 and ca 125 tumour markers evaluated from an international quality assessment survey.

Data collected in the 1993 and 1994 cycles of an international External Quality Assessment (EQA) programme were cumulatively analysed to evaluate the analytical performance of the methods currently in use for routine assay of mucinous tumour markers CA 19-9, CA 15-3 and CA 125. On average the between-laboratory variability was 14.7 and 15.8 CV% for CA 15-3 and CA 125 respectively. For CA 19-9, a markedly worse between-laboratory variability (on average 27.2 CV%) was found; the agreement of CA 19-9 results worsened in the last few years when new isotopic techniques became available. The variability component attributable to systematic differences between methods/kits was relatively small for CA 15-3 and CA 125 (17% and 21% of the total variability), while it was markedly larger for CA 19-9 (45% of the total variability). The precision of the methods/kits most often used in the survey ranged from 9.6 to 13.9 CV% for CA 125 and from 10.8 to 14.1 CV% for CA 15-3. For these two tumour markers the precision of the traditional IRMAs does not appear to be different from that of the new fully automated non-isotopic techniques. The precision of CA 19-9 methods was on average worse from (11.9 to 19.2 CV%), even though the precision of the two automated systems was better than that of IRMAs. In conclusion, the results of this study indicate that the between-laboratory agreement for CA 15-3 and CA 125 assays appears satisfactory while the CA 19-9 assay shows larger differences between methods and is affected by poorer precision of kits.

Hormonal, water balance, and electrolyte changes during sixty-day confinement.

The EXEMSI experiment has made it clear that it is difficult to perform psychological and physiological protocols satisfactorily in the same study. It is, therefore, essential that the objectives of study be defined clearly before the start. While behavioral and psychological studies may be possible and provide valid results for a small group of mixed gender, it is more difficult to conduct valid physiological studies due to large differences between individuals and even in the same individual over time. As stated before, it is unusual in space research on humans and even during space simulation studies to have large and homogeneous groups of subjects. The consequence is that the results remain tentative. For a better understanding of the physiological data collected during the ISEMSI ad EXEMSI experiments, they should be correlated with the results of the psychological studies. One of the conclusions drawn from the ISEMSI experiment was that confinement provides a valuable parallel to other simulations of weightlessness, such as bedrest. The same pattern of changes in parameters like the blood volume regulating hormones renin and aldosterone was observed as in bedrest. After the EXEMSI study we can say that the conditions imposed by confinement, high work load, and stress, potentiate these effects. This implies that in using head-down bedrest as a weightlessness simulation the confinement effects must be identified by setting adequate control conditions for the head-down position, for short-term as well as for long-term simulations. Indeed, we have seen in the two isolation studies that confinement may have its effects at the beginning of the isolation period (EXEMSI) as well as during the entire isolation period (ISEMSI). In planning for EXEMSI we wanted to obtain more insight in some of the phenomena observed during ISEMSI by the introduction of new techniques such as the doubly labeled water method for determination of total body water. However, in some cases the opposite effects of those encountered in ISEMSI were found. This was probably due to the many changes in the experimental scenario, like number of subjects, mixed gender, living space per subject, and workload. Thus, for future isolation studies the operational scenario should be better examined and preferably standardized. Nevertheless, in such studies as well as in long-term sojourns in a space station, the crew size will not be larger than that of the EXEMSI crew. Physiologists will, therefore, have to become familiar with the study of small groups of subjects and to try to overcome the problems of large individual differences and statistical analysis of data from small groups.

Comparison of immunoassays for tumor markers CA 19-9, CA 15-3 and CA 125: data from an international quality assessment scheme.

Data collected in the 1993 and 1994 cycles of an international external quality assessment (EQA) program and in a national multicenter collaborative study were cumulatively analyzed to evaluate the standardization of the methods currently in use for the assay of mucinous tumor markers CA 19-9, CA 15-3 and CA 125. On average the between-laboratory variability was 15.2 and 16.0 CV% for CA 15-3 and CA 125 respectively; the between-laboratory variability found for CA 19-9 was markedly worse (mean 28.3 CV%). The variability component attributable to systematic differences between different methods/kits was relatively small for CA 15-3 and CA 125 (18% and 24% of the total variability) but markedly larger for CA 19-9 (48% of the total variability). The agreement of CA 19-9 results worsened in the last few years when new nonisotopic techniques became available. The precision of the methods/kits most used in the survey ranged from 9.9 to 13.3 CV% for CA 125 and from 11.6 to 13.9 CV% for CA 15-3. For these two tumor markers the precision of the traditional IRMAs does not appear different from that of the new fully automated nonisotopic techniques. The precision of CA 19-9 methods was on average worse (from 11.7 to 19.6 CV%) although two automated systems exhibited a precision better than that of IRMAs. In conclusion, the results of this study indicate that CA 15-3 and CA 125 are satisfactorily assayed whereas CA 19-9 assay appears affected by larger differences between methods and by poorer precision of laboratories and kits.

Pharmacodynamics and relative bioavailability of cabergoline tablets vs solution in healthy volunteers.

The effect of formulation on the urinary pharmacokinetics, pharmacodynamics, and relative bioavailability of cabergoline was investigated. Twelve healthy female volunteers, aged 23-35 years, were treated, according to an open, randomized, crossover design, with cabergoline (1-mg single oral dose) both as tablets and as a solution. The two administrations were separated by a 4-week wash-out period. Cabergoline and prolactin were measured in urine and plasma, respectively, by specific radioimmunoassays. Blood samples were collected before and up to 30 days after dosing. Urine was collected before and up to 8 days after dosing. Cabergoline elimination half-lives calculated from urinary data were 68 and 63 h after administration of the tablets and the solution, respectively. Urinary excretion of unchanged cabergoline accounted, on average, for 1.92% (range, 0.14-3.26) and 1.80% (range, 0.67-3.09) of the dose after administration of the tablets and the aqueous solution, respectively. Relative bioavailability of tablets vs solution was 99% (geometric mean with the 90% confidence intervals of 68-144%). Prolactin levels in 10 out of 12 subjects fell below the detection limit of the assay (1.5 micrograms/L) after both treatments. The mean maximum prolactin decrease (ca. 70%) was achieved by 2 or 3 h after dosing; the effect persisted up to 9 days, being completely exhausted 23-28 days after dosing. The analysis of variance performed on the pharmacodynamic effects of the two cabergoline formulations indicated that the percent decreases of plasma prolactin levels were not significantly different for tablets and solution. These results indicate that the pharmacodynamics and relative bioavailability of cabergoline are not influenced by formulation, as tablets or solution.

'Oncocheck': an international external quality assessment scheme for immunoassays of tumor markers.

Starting from November 1990, an international External Quality Assessment Scheme (EQAS) for immmunoassays of tumor markers has been organized. Presently, 238 laboratories from France, Germany, Italy, Japan and Spain participate in the scheme. In this report the main features of the EQAS and data processing are outlined. Results collected during the 1992-cycle allow evaluation of the state of the art of AFP, CEA, CA 19-9, CA 15-3, CA 125 and PSA immunoassays. According to their analytical performances, the 6 tumor marker immunoassays can be classified into several groups, the first including AFP and CA 15-3 for which both total variability and within-kit agreement are good. For CEA assay, performance can be considered as satisfactory even though further improvements of between-lab agreement would be welcome. For the 3 other tumor markers, the higher total variability indicates an urgent need for a better standardization by improvement of either both within-kit and between-kit agreements (CA 19-9) or between-kit agreement mainly (PSA, CA 125).

European isolation and confinement study. Blood pressure, volume-regulating hormones, and electrolytes.

In the ISEMSI confinement experiment we observed three main reactions concerning blood volume regulation: first, a stress activation produced by the high workload that the subjects had to accomplish, both before the isolation period and during the two first weeks of isolation; second, a typical defense reaction to the environment occurred, at least during the two initial days of isolation and later perhaps between crew members themselves; and third, a change in blood volume regulating hormone levels may have been the result of the increase of sympathetic system activity and of dehydration. Confinement is certainly a good and valid method to simulate space station life (with the absence of weightlessness). So, it is interesting to use these confinement studies to perform biological and physiological experiments to obtain greater knowledge, but also to prepare and validate new scientific protocols or new scientific equipment suitable for spaceflights. Even more so, confinement is a good way to prepare and train crew members for long-term space missions. It would seem desirable to confirm the scientific results obtained during the ISEMSI campaign, and especially those for the regulation of the blood volume. This will be done during the next confinement experiment, called EXEMSI, in which four subjects will be subjected to 60 days of confinement.

[Control program of interlaboratory quality for the assay of cyclosporine in biological fluids]. / Programme de contrôle de qualité interlaboratoire pour le dosage de la ciclosporine dans les liquides biologiques.

The following article deals with the quality control program proposed since 1988 to the French laboratories performing the measurement of cyclosporine concentration in whole blood or in plasma. First, we describe how the program was organized and its evolution over time. Second, we discuss the main results generated. Those results enable us to state positively that the methods in use at present in the laboratories are precise and give highly reproducible results.

Pharmacokinetics of cyclosporine in hyperlipidaemic long-term survivors of heart transplantation. Lack of interaction with the lipid-lowering agent, fenofibrate.

Cyclosporine (Cy) binds to lipoproteins in plasma. In order to test if its pharmacokinetics would be modified when efficient lipid-lowering treatment is introduced, a study has been done of Cy pharmacokinetics and any interaction with the lipid-lowering agent fenofibrate in hyperlipidaemic long-term, survivors of heart transplantation. Fenofibrate 200 mg once daily significantly reduced blood lipids (cholesterol 6.5 vs 7.7 mmol/l; apoprotein B 1.2 vs 1.6 g/l) but did not modify mean whole blood Cy trough levels (113 before fenofibrate vs 103 ng.ml-1), Cmax (812 ng.ml-1 by RIA and 757 ng.ml-1 by HPLC before fenofibrate versus 865 and 741 respectively, during fenofibrate); tmax (1.6 and 1.7 h before fenofibrate versus 1.4 and 1.4 h respectively), and t1/2 (13.9 and 11.1 h versus 9.5 and 10.7 h). The only adverse effect was an increase in creatinine (157 vs 145 mmol/l). Further studies are needed to investigate the mechanism of Cy-fenofibrate nephrotoxicity and to evaluate the long-term efficiency and safety of fenofibrate after heart transplantation.

Immunoassay external quality assessment schemes in France.

Several immunoassay external quality assessment schemes (EQAS) are currently operating in France. Three of them are described, special reference being made to their philosophy, aims, structure and organisation. The results obtained since the EQAS were set up, enable us to appreciate the state of the art of immunoassays and its evolution with time. The results, also, point out the need to use control samples of high quality.

Antigravity suit inflation: kidney function and cardiovascular and hormonal responses in men.

To investigate the effects of lower body positive pressure (LBPP) on kidney function while controlling certain cardiovascular and endocrine responses, seven men [35 +/- 2 (SE) yr] underwent 30 min of sitting and then 4.5 h of 70 degrees head-up tilt. An antigravity suit was applied (60 Torr legs, 30 Torr abdomen) during the last 3 h of tilt. A similar noninflation experiment was conducted where the suited subjects were tilted for 3.5 h. To provide adequate urine flow, the subjects were hydrated during the course of both experiments. Immediately after inflation, mean arterial pressure increased by 8 +/- 3 Torr and pulse rate decreased by 16 +/- 3 beats/min. Plasma renin activity and aldosterone were maximally suppressed (P less than 0.05) after 2.5 h of inflation. Plasma vasopressin decreased by 40-50% (P less than 0.05) and plasma sodium and potassium remained unchanged during both experiments. Glomerular filtration rate was not increased significantly by inflation, whereas inflation induced marked increases (P less than 0.05) in effective renal plasma flow (ERPF), urine flow, osmolar and free water clearances, and total and fractional sodium excretion. No such changes occurred during control. Thus, LBPP induces 1) a significant increase in ERPF and 2) significant changes in kidney excretory patterns similar to those observed during water immersion or the early phase of bed rest, situations that also result in central vascular volume expansion.