[Relation between physical activity, muscle function and IGF-1, testosterone and DHEAS concentrations in the elderly]. / Relations entre l'activité physique, la fonction musculaire et les concentrations en IGF-1, testostérone et DHEAS lors de l'avancée en âge.

OBJECTIVE: Lower amounts of circulating anabolic hormones are thought to accelerate the age related decline in muscle mass and function. Replacement therapies are promising interventions but there are problems with these therapies. Thus alternative strategies should be developed. The age related changes in hormonal status may be probably influenced by exercise. The purpose of this study was: a) to confirm with other methods, more adapted for elderly people, the results of a previous study that has shown relationship between physical activity (PA) and quadriceps muscle function with dehydroepiandrosterone sulphate (DHEAS), insulin like growth factor-1 (IGF-1). Quadriceps muscle power (Pmax) is measured in this new work with a recently developed leg extensor machine and, b) to complete the results of the first study examining simultaneously the relationship between PA, Pmax and cardiorespiratory fitness (VO2max) with DHEAS, IGF-1 and testosterone in a group of healthy elderly people. METHODS: Fifty independent, community dwelling elderly subjects (25 mens and 25 womens) aged from 66 to 84 volunteered to participate in the study. PA was evaluated by the questionnaire and expressed using two activity indices: mean habitual daily energy expenditure (MHDEE) and the daily energy expenditure corresponding to leisure time sports activities (Sports Activity). Pmax and optimal shortening velocity (vopt) were measured on a Ergopower dynamometer. The Pmax was expressed relative to body mass, Pmax/kg (W kg-1), and relative to the mass of the two quadriceps muscles, Pmax/Quadr (W.kgQuadr-1). VO2max has been measured during a maximal treadmill exercise. RESULTS: In women, IGF-1 correlated significantly with MHDEE (r = 0.54, P = 0.004), Pmax/kg (r = 0.54, P = 0.004) and Pmax/Quadr (r = 0.46, P = 0.02), whereas DHEAS with MHDEE (r = 0.54, P = 0.004), Sports Activity (r = 0.65, P < 0.001), VO2max (r = 0.46, P = 0.02), Pmax/kg (r = 0.46, P = 0.02) and Pmax/Quadr (r = 0.55, P = 0.004). No such correlation was found in men. CONCLUSION: These findings confirm that in healthy elderly women physical activity, cardiorespiratory fitness and quadriceps muscle function are similarly related to levels of circulating DHEAS and IGF-1 suggesting a favourable influence of exercise on anabolic hormonal production in the elderly.

[Biochemical diagnosis of pheochromocytoma and neuroblastomas]. / Diagnostic biochinique du phéochromocytome et du neuroblastome.

Pheochromocytoma and neuroblastoma are distinct tumours, but their biological diagnosis is based on secretion increase of one or several catecholamines. Assays have to be very sensible and specific for an early diagnosis. 24 hours urinary catecholamines and metabolites are currently measured, but technical improvements permit plasma metanephrine assay, an excellent indicator of pheochromocytoma. HPLC coupled to electrochemical detection represents the most efficient methodology. After a review of urinary and plasma assay methods, the authors show usual values of catecholamines, metanephrines, HVA and VMA, according to ages, and give examples of results encountered in classical or not tumours and in falsely positive cases. Urinary metanephrine assay is the most sensible and specific in biological diagnosis of pheochromocytoma, while catecholamines and VMA assays lack of sensibility. Results have to be given by 24 hours and by creatinine ratio. Metanephrine assay can be performed also in plasma and exhibits the same interest. However, in urine as in plasma, in case of renal failure, results cannot be interpreted. Neuroblastoma biological diagnosis is based classically on HVA, VMA, and dopamine assays, nowadays only in 24 hours urine (or in urinary micturition for screening), and results are also expressed as creatinine ratio. But even if several assays are advisable, 5% of the neuroblastoma cases do not produce increased catecholamine values. In some cases, metanephrine assay could be of interest. After the age of 12 months, clinical expression of neuroblastoma is dramatic in 70% of cases. So, a biological screening has been experimented in several countries including France. A French translation of the consensus conference report (1998) is appended, which shows the complexity of neuroblastoma screening. Now, there is no evidence that early tumour detection by screening lessens the mortality rate, but a weak benefit is not excluded.

Comparison of progesterone concentration determination by 12 non-isotopic immunoassays and gas chromatography/mass spectrometry in 99 human serum samples.

A single serum progesterone determination may be highly predictive for early pregnancy and in vitro fertilisation and embryo-transfer outcomes. We therefore compared 12 direct non-isotopic progesterone immunoassays with gas-chromatography/mass spectrometry (GC/MS). For each assay, data from the analysis of 99 individual sera were compared with data obtained by GC/MS, using regression and bias plot analyses and the ratio method. We observed a larger difference in concentration between high and low values and a broader distribution of results for immunoassays than for GC/MS. All immunoassays displayed bias in the calibration process and a lack of specificity and/or sensitivity, to various degrees. We tried to identify the parameters of the assay procedure that might contribute to these discrepancies. None of the criteria investigated (antibodies, control and preparation of calibrators, blocking agents and choice of tracer) had a significant effect when studied alone.

Evaluation of the rennin-angiotensin-aldosterone system in hypertensive Ivoirian subjects--preliminary results.

Experimental data on the renin-angiotensin-aldosterone system in black Africans are very limited. In this study we set up our own values (plasma renin and aldosterone) in a group of normotensive Ivoirian male and female (N = 43) subjects and compared our results with those published for the white population. The values were approximately the same. A parallel study was carried out in a black population (N = 48 men and women) consulting a doctor for the first time for hypertension in different regions of Côte d'Ivoire. The mean values of plasma renin and aldosterone were significantly higher (P < 0.05) in the hypertensive compared with the control group.

Lack of correlation between 6-month fluctuations in habitual physical activity and testosterone. Concentrations in elderly subjects.

BACKGROUND: To examine the influence of long-term modifications in habitual physical activity (PA) on resting total testosterone (TT) concentrations in a group of healthy elderly subjects. EXPERIMENTAL DESIGN: 6-month prospective study. SETTING: University research departments of exercise physiology and geriatric medicine. PARTICIPANTS: thirty-one independent, community dwelling elderly subjects (16 men and 15 women) aged > 65 who volunteered to participate. INTERVENTIONS: correlational study. MEASURES: PA was evaluated by a questionnaire QAPSE (Questionnaire d Activité Physique Saint-Etienne) and expressed by two activity indices: mean habitual daily energy expenditure (MHDEE) and daily energy expenditure corresponding to leisure time sports activities (Sports activity index). RESULTS: Changes in resting TT concentrations were not related to changes in MHDEE (r = -0.28; p = 0.30 and r = -0.001; p=0.99) and Sports activity (r = -0.30; p = 0.26 and r = -0.05; p = 0.85) in men and women, respectively. Similarly, relative (%) changes in TT were not related to relative changes in MHDEE (r = -0.16; p=0.55 and r = 0.17; p=0.55) and Sports activity (r = -0.11; p=0.68 and r = 0.02; p = 0.92). CONCLUSIONS: The levels of habitual PA undertaken by an average senior do not appear to affect TT levels in healthy active elderly men and women.

Determination of plasma methoxyamines.

The determination of plasma methoxyamines is the most informative parameter for the diagnosis of pheochromocytoma. Very good sensitivity and specificity are necessary for this. The measurement of deconjugated (free plus sulfate-conjugated) metanephrines is more useful. Concentrations are 10-fold higher than free metanephrine concentrations. The methodology generally uses a column purification and high performance liquid chromatography with electrochemical detection. We have tested six commercial columns used for the purification step. We chose a rapid and reliable method with a mixture of strong cation-exchange and strong anion-exchange groups bonded onto a silica column. This protocol has been validated with samples of plasma from normal subjects, healthy elderly people, renal failure patients and patients with pheochromocytoma.

Leg extensor power and dehydroepiandrosterone sulfate, insulin-like growth factor-I and testosterone in healthy active elderly people.

We examined the association between quadriceps muscle function and serum levels of dehydroepiandrosterone sulphate (DHEAS), insulin-like growth factor I (IGF-I) and testosterone in a group of healthy elderly people. Fifty-three independent, community-dwelling elderly subjects (26 men and 27 women) aged from 66 to 84 years volunteered to participate in the study. Physical activity (PA) was evaluated by a questionnaire. Quadriceps maximal muscle power (Wmax) and optimal shortening velocity (v(opt)) were measured on a friction-loaded non-isokinetic cycle ergometer. The Wmax is expressed in relation to body mass (W(max/kg), W x kg(-1)), and in relation to the mass of the two quadriceps muscles (W(max/Quadr), W x kg(Quadr(-1))). In women, when adjusted for age, anthropometric measurements and PA indices, IGF-I correlated significantly with W(max/kg) (partial correlation: r = 0.59; P = 0.001), W(max/Quadr) (r = 0.58; P = 0.002) and v(opt) (r = 0.53; P = 0.004), whereas DHEAS was correlated significantly with W(max/kg) (r = 0.54; P = 0.003) and W(max/Quadr) (r = 0.58; P = 0.002). No such correlation was found in men. These findings indicate that in healthy elderly women lower values for quadriceps muscle Wmax and v(opt) are related, independently of age, anthropometric measurements and PA indices, to lower circulating levels of DHEAS and IGF-I.

[Validity of immunochemical methods for bloody progesterone: a study performed in 1998]. / Validité des dosages immunochimiques de la progestérone dans le sang: étude réalisée en 1998.

The aim of this study was to compare, with manufacturer's agreement, twelve direct and non-isotopic commercial assays of progesterone. We have evaluated the analytical performances: low limit detection, imprecision, accuracy (recovery and dilution tests) and we have tested some patient samples. Results were compared to a reference method using isotope dilution Gas Chromatography and Mass Spectrometry combination (GC-MS). For each assay, analytical qualities and defaults are established. Large differences are found between progesterone concentration measured on the same sample with the different methods essentially for the low concentrations. Comparison with GC-MS raised questions about the accuracy of the different assays. This work will be aid laboratories to their choice and/or validation.

Influence of acute and chronic exercise on insulin-like growth factor-I in healthy active elderly men and women.

We examined the influence of physical activity (PA) and acute resistance exercise on insulin-like growth factor I (IGF-I) in 39 healthy, independent, community-dwelling elderly volunteers (14 men and 25 women) aged from 66 to 84. In a longitudinal non-interventional study, we compared the fluctuations in IGF-I levels and changes in habitual PA over a 6-month period. PA was evaluated by a questionnaire QAPSE (Questionnaire d'Activité Physique Saint-Etienne), and expressed by two activity indices: mean habitual daily energy expenditure (MHDEE), and daily energy expenditure corresponding to leisure time sports activities (sports activity index). In an experimental design we compared IGF-I levels before, at 1 and 15 minutes after short quadriceps muscle exertion (10 consecutive repetitions) carried out with optimal muscle power development. Changes in IGF-I levels over a 6-month period were positively related to changes in MHDEE (r = 0.41; p<0.01), and sports activity (r = 0.40; p<0.02). When analyzed by gender, these correlations were still statistically significant in women (r = 0.39; p = 0.05 and r = 0.41; p<0.05), but not in men (r = 0.36; p = 0.21 and r = 0.20; p = 0.50). No changes (p = 0.17) were observed in serum IGF-I concentrations after the acute exercise protocol. These findings indicate that IGF-I levels are positively related to fluctuations in habitual PA, at least in women. Further research is needed to establish the best intensity and duration of exercise training to induce these changes.

Physical activity and dehydroepiandrosterone sulphate, insulin-like growth factor I and testosterone in healthy active elderly people.

OBJECTIVE: to examine the association of physical activity and cardio-respiratory fitness with dehydroepiandrosterone sulphate (DHEAS), insulin-like growth factor I (IGF-I) and testosterone in healthy elderly people. DESIGN: cross-sectional study. SETTING: university research department and department of geriatric medicine. PARTICIPANTS: 60 independent, community-dwelling elderly subjects (26 men and 34 women) aged 66-84 who volunteered to participate. MEASUREMENTS: physical activity was evaluated by the Questionnaire d'Activité Physique Saint-Etienne and expressed by three indices: mean habitual daily energy expenditure (MHDEE), daily energy expenditure (DEE) [comprising activities with intensities corresponding to at least three metabolic equivalents (MET; 3.5 ml.kg1 x min1 of oxygen consumption)] and sport activity. Cardio-respiratory fitness was expressed by maximal oxygen consumption (VO2max). RESULTS: In women, DHEAS correlated with VO2max (partial correlation: r=0.33; P=0.05), MHDEE (r=0.50; P=0.002), DEE > 3 METs (r=0.49; P=0.003) and sport activity (r=0.35; P=0.04) whereas IGF-I correlated with MHDEE (r=0.48; P=0.004). DHEAS was correlated with IGF-I (r=0.43; P < 0.02) and with testosterone (r=0.41; P < 0.02). No such correlation was found in men. CONCLUSION: lower habitual physical activity is related to lower levels of circulating DHEAS and IGF-I independently of age and anthropometric measures. Lower maximal aerobic capacity is associated with lower DHEAS concentrations, in healthy elderly women.

[Pregnancy and perinatality: biological follow-up]. / Grossesse et périnatalité: suivi biologique.

Blood and urinary tests which are necessary for pregnancy diagnosis and follow-up, for newborn and mother medical supervision, during the month following birthday, are today described in reglementary texts, laws, and recommendations such as advised medical references (RMO). These documents specify the nature of obligatory tests, the checking rhythm and the list of useless tests. hCG research remains necessary for pregnancy diagnosis, but hCG dosage is essential only in case of programmed medical assistance or pathological pregnancy (extrauterine pregnancy, hydatiform mole, choriocarcinoma). The obligatory follow-up of a pregnant woman includes determination of blood groups, research of infectious agents responsible for diseases (toxoplasmosis, rubeola, hepatitis B, syphilis), proteinuria and glycosuria research and blood count according to a given calendar. When the mother's condition is bad and reminiscent of a pathological pregnancy, when a genetic risk exists for the fetus or when fetal growth is abnormal as indicated by echographic control (intra-uterine growth retardation), laboratory tests are used to follow the maternal pathological course (arterial hypertension, diabetes mellitus, anemia, bacterial, viral or parasitic disease), to verify the existence of a genetic disease, to know about the fetal functional state (by amniocentesis or cordocentesis), to identify an erythrocyte fetomaternal incompatibility. Since last trimester pregnancy accidents are able to endanger mother's and fetus lives, the feto-maternal follow-up must be adjusted to pathological diagnosis types and requires a particular supervision of the delivery. Finally mother and child must undergo a post-natal follow-up during the four weeks after birthday (perinatality control).

Convenient chromatographic prepurification step before measurement of urinary cortisol by radioimmunoassay.

We applied various prepurification protocols (extraction with different solvents, liquid/solid separation on bonded silica media, Celite, and Sephadex LH20 chromatography) with a range of commercially available RIA kits to measure cortisol in urine samples. We then compared the results with the concentrations measured by a HPLC method validated with reference to isotope dilution gas chromatography-mass spectrometry. We conclude that chromatography on a commercial, prepacked diol minicolumn (Waters Sep-Pak Vac RC) in combination with dichloromethane extraction is a convenient and very effective purification step before RIA of urinary cortisol in patients not receiving corticoid medication. We tested numerous steroids for interference and found that free polar cortisol derivatives (hydroxylated or hydrogenated) could only partially account for the overestimations routinely encountered when free urinary cortisol concentrations are measured by direct RIA.

[Measurement of biological activity of hLH. Multicenter study]. / Measure de l'activité biologique de la hLH. Etude multicentrique.

This report describes the results of a multicentric study of biological methods for luteinizing hormone (hLH). The production of testosterone by animal Leydig cells is used by five laboratories with different methodologies including rat, mouse and pig cells. Dose-response curves of testosterone production, quality criteria and results on a physiological population of men and women, are reported and discussed. It is concluded that the bioactive determination of hLH must be considered as a reference method used in discordances between clinic and immunological methods.

European collaborative study of LH assay: 3. relationship of immunological reactivity, biological activity and charge of human luteinizing hormone.

This report describes the results of the third part of the collaborative study organized by a working group sponsored by the Community Bureau of Reference of the European Community Commission. The aim of the present work was to establish the link between immunoreactivity and biological activity of human LH, thus allowing to determine the antigenic domains of the molecule involved in the induction of the biological effect. The relationship between immunoreactivity and electric charge of hLH was also studied. This work allowed to further apprehend hLH isomorphism and its role in discrepancies observed among hLH assays and clinical status. It also made the feasibility of measuring biologically active isoforms by an immunological method to be assessed. The effect of 36 mAb with known epitopic specificity, was evaluated on both hLH binding to rat membrane receptor and hLH induced production of testosterone by porcine Leydig cells. All the epitopes located on the beta subunit were found to be strongly involved in the biological activity whereas 4/9 and 10/18 epitopes present on the alpha subunit or specific for the holomolecule respectively appeared weakly involved. Assaying biological hLH using immunological method would require that mAb specific for all the epitopes involved in the receptor activation be tested, and thus appears presently unsuitable for routine clinical evaluation. In the previous work some LH immunoassays were found to underestimate LH concentrations (J. Endocrinol. Invest 1994, 17: 397-406 and 407-416). The mAb used in liquid phase in these kits were found in the present work to be directed against the domains of LH weakly involved in the activation of the receptor and would suggest that bioactive LH would be misevaluated by these kits. The immunoreactivity of hLH isoforms separated by isoelectric focusing (IEF) in liquid phase was also determined. IEF allowed to separate three groups of hLH isoforms but none of them exhibited a specific discriminating pattern of immunoreactivity when they were tested against a panel of mAb. It suggests that, in our experimental conditions, the electric charge and the immunoreactivity of hLH were not closely linked.

[Measurement of salivary cortisol with four commercial kits]. / Mesure du cortisol salivaire par quatre trousses du commerce.

Salivary cortisol was determined with four commercially available immunoassays: one enzyme-immunoassay (Cortisol Biotrol), two fluoro-immunoassays (TDX, Abbott; Stratus, Baxter) and one radioimmunoassay (Coat-A-Count, Behring). In order to improve the sensitivity of these immunoassays, it was necessary to increase sample volumes. Thus an extraction step had to be included in the procedure. It was performed with either methylene chloride or Bond Elut. However, the Coat-A-Count kit may be applied directly on salivary aliquots. The results obtained were compared to those yielded by the reference technique which includes a chromatographic step on Sephadex LH 20. According to the present data (n = 20) no kit appears to be adequate for salivary cortisol measurement at any level, at least in the conditions applied in this study. However, the introduction of a chromatographic step in the procedure improved greatly the specificity. Nevertheless, the best results were obtained with the Coat-A-Count kit. Indeed, they were generally similar to those of the reference technique, but some discrepancies were observed at low levels.

[Validity of immunochemical assays for blood estradiol. Study conducted in 1994]. / Validité des dosages immunochimiques de l'estradiol sanguin. Etude réalisée en 1994.

Since 1989 many kits have been commercialized for measuring estradiol by non-isotopic immunoassays which largely contributed to the diffusion of this parameter. Rapidly, in quality control schemes, a wide dispersion of results has been observed. In this paper, the kits' characteristics (precision, accuracy and detection limit) have been studied and compared to a reference method (gas chromatography-mass spectrometry). In addition, estradiol determinations in patients with various pathophysiological situations have been performed to underline the difficulties of interpretation of this analyte when clinicians compare results given by the different kits. This work demonstrates that precision and accuracy varied both with the kit and the level of concentration of estradiol and gives better results for high concentrations of estradiol. In seven out of the 12 kits, the limits of detection are in good agreement with those announced by manufacturers. On the contrary, the determination on patients sera gives results with a high variability with no systematic error.

[Value of immunoassay of hCG for the determination of high plasma concentrations]. / Validité des immunodosages de l'hCG appliqués à la détermination de concentrations plasmatiques élevées.

The serum concentration of human chorionic gonadotrophin is higher in trisomy 21 than in normal pregnancy. Our study aimed at comparing 17 commercial kits, both those for measuring high concentrations and others for diagnosis and initial follow-up of pregnancy under specific conditions, ie for measuring hCG concentrations between weeks 14 and 18 of pregnancy. All kits were satisfactory; however, usual values for the gestational weeks concerned need to be established for each reagent.

[Recommended methods for the determination of catecholamines and their metabolites in urine. Significance of results in the diagnosis and follow-up of pheochromocytoma and neuroblastoma]. / Méthodes recommandées pour le dosage des catécholamines et de leurs métabolites dans l'urine. Signification des résultats dans le diagnostic et le suivi des phéochromocytomes et des neuroblastomes.

Laboratory investigation of catecholamines is useful for the clinician in the diagnosis and management of phaeochromocytoma and neuroblastoma. This work summarizes current knowledge on catecholamines and their metabolites and discusses the main indications for their determination. It also examines the most practical methods for studying the secretion of these hormones, ie extraction, separation and quantification using high performance liquid chromatography coupled with electrochemical detection. The workshops attended by the members of the catecholamines working party of the French Clinical Biology Society and phaeochromocytoma and neuroblastoma specialists enabled the role of such determinations in the diagnosis and management of these diseases to be clarified.

[Immunoassay and measurement of the biological activity of serum hLH: results obtained of a multicenter study]. / Immunodosages et mesure de l'activité biologique de l'hLH sérique: résultats obtenus lors d'une étude multicentrique.

The discrepancy between results obtained using different anti-hLH antibodies available on the market led us (in the context of a contract with the CNAMTS) to compare the results of hLH immunoassays (eight different reagents) with those of hLH bioassays based on the measurement of the steroidal activity of this hormone on mouse, rat or pig testicular preparations (five methodologies). Twenty-nine samples of serum or human serum pools were tested. The values resulting from these tests reveal differences linked to the nature of the antibodies or to the methodological conditions used for the testicular preparations. Some antibodies seem unable to identify some hLH forms which, however, can be detected by biological methods.

Determination of urinary cortisol with three commercial immunoassays.

Urinary cortisol determination was performed with three commercially available immunoassays: one enzyme-immunoassay (Cortisol Biotrol) (EIA) and two radioimmunoassays: Quanticoat Cortisol (Kallestad Diagnostics) (KD-RIA) and GammaCoat Cortisol (Clinical Assays) (CA-RIA). Four procedures were carried out. Procedure I (methylene chloride extraction) was applied to EIA and CA-RIA and procedure II (ethyl acetate extraction) to KD-RIA. Procedure III combining procedure I and column chromatography on Sephadex LH 20 in methylene chloride was applied to the three kits. Procedure IV consisting of carbon tetrachloride preextraction and extraction with cyclohexane-ethyl acetate (50:50, v/v) was applied to CA-RIA. The results obtained were compared with those of the reference technique, "on-line" HPLC with u.v. detection. Two groups of results were arbitrarily considered, those below (n = 28) and those above (n = 6) 270 nmol/l. In the first group, the results were markedly overestimated when the procedure was limited to solvent extraction. Conversely, the third procedure proved the efficiency of the chromatographic step since specificity was greatly improved in the three cases, the levels obtained with either kits being similar to those of the reference technique. The second group of results (above 270 nmol/l) yielded by the three kits were not always higher than those of HPLC when the procedure was limited to solvent extraction. When column chromatography was included in the procedure, the results were comparable to those of HPLC in three cases and lower in the three others. Since, the latter samples were collected after cortisol administration, and overestimated cortisol values obtained by HPLC might be due to the interference of some cortisol metabolites.