[A drift induced by the acquisition of a video system linked to a microscope]. / Une dérive induite par l'acquisition d'un système vidéo affilié à un microscope.

Further to the acquisition of a new material consisting in a microscope and its video equipment, we report the example of some drift with regards to written, updated and validated procedures. Indeed, once the new material was fully operating, depending on the operator, the (spermocytogrammes) morphology was realized either in oculars or by using the video screen. The statistical study performed on the results from 50 patients showed a significant difference in the results of the appreciation of the microcephalous and the macrocephalous according to the mode of observation that had been used. As an outcome of this study, we insisted that everyone within the team should work consistently and comply with the current procedures.

[Spermatozoon search in forensic medicine]. / La recherche de spermatozoïdes sur réquisition.

The search for sperm is part of the examinations wich may be prescribed, following a request of jucicial services, in case of a suspicion of sexual assault. This article applies to detail how the different phases, pre-analytical, analytical and post-analytical, must be absolutely adhered to so that the biologist, expert or not, in charge of this analysis, can be deliver results (positive or negative) that cannot be challenged.

[Tobacco: a potential inductor of lipid peroxidation of the human spermatozoon membrane?]. / Le tabac: un inducteur potentiel de peroxydation lipidique membranaire du spermatozoïde humain?

OBJECTIVE: To determine the membrane lipid peroxidation of human spermatozoon in a cohort of smokers in comparison of never-smokers. MATERIALS AND METHODS: Malondialdehyde (MDA), a stable product of the membrane lipid peroxidation, was assessed in 25 smokers and in 17 never-smokers. In parallel, an evaluation of sperm characteristics was realized for all the studied patients. RESULTS: For the first time, between smokers and never-smokers, a significative increase of MDA concentrations was found by the U-Mann and Whitney test (0.118 +/- 0.176 vs 0.0392 +/- 0.0117 nmol/10(6) spermatozoa), a decrease of the forward motility (grade A), (18 +/- 8 vs 25 +/- 8%) and total sperm count (265.56 +/- 186.96 x 10(6) vs 399.30 +/- 322.23 x10(6)), and also an increase of tapering heads (6 +/- 4 vs 2 +/- 2%) or morphological stress pattern cells (39 +/- 6 vs 24 +/- 5%). In the smokers group, negative significative correlations were found by the non-parametric Spearman test between the MDA concentrations and the sperm count per mL (r=-0.767, p<0.001), the total sperm count (r=-0.656, p<0.001) and the percentage of normal morphology (r=-0.644, p<0.001). CONCLUSIONS: Given of deleterious effects of tobacco in a large panel of human cells and specially on the male gametes, the increase of spermatozoon membrane MDA concentrations and the sperm abnormalities found in the group of smokers may be linked to cigarette smoking.

Hyaluronidase activity in serum of patients with monoclonal gammapathy.

Hyaluronidase and hyaluronic acid, two substances thought to be strongly implicated in carcinogenesis, were assessed in the plasma of 35 patients with newly documented monoclonal gammapathy and in 25 control patients. A significant increase was found in plasma hyaluronidase activity in the patients with monoclonal gammapathy. A statistically significant positive correlation was found between hyaluronidase activity and monoclonal immunoglobulin levels in plasma. An increase in serum hyaluronidase activities may be a response to the deleterious effect of hyaluronic acid in cell migration and tumor progression. Further studies are needed to assess the value of hyaluronidase activity as a marker of tumor progression.

[Lipid peroxidation, morphological stress pattern and nuclear maturity of spermatozoon]. / Peroxydation lipidique, profil morphologique de stress testiculaire et maturité nucléaire des spermatozoïdes.

Malondialdehyde, a stable lipid peroxidation product of polyunsaturated fatty acids, was assessed in sperm and seminal plasma of 21 patients to test a potential link between this compound and, the percentage of spermatozoon exhibiting a morphological stress pattern, and the results of the nuclear maturity test. An analysis by the Spearman rank correlation shows a statistically significant positive correlation between the sperm malondialdehyde levels and the percentage of spermatozoon with morphological stress pattern (r = 0.505, p = 0.024). No correlation was found between the sperm malondialdehyde concentrations and the results of nuclear maturity test. The result of this work shows that spontaneous stress pattern seems to be linked, in part, to an excessive lipid peroxidation of the spermatozoon membrane. On the other hand, it seems that malondialdehyde have no effect on the chromatin maturity. According to the deleterious effects of lipid peroxidation on spermatozoon, the determination of both percentage of stress pattern and sperm malondialdehyde levels seems to provide useful information on the fertility status of patients.

Seminal androgen concentrations and residual sperm cytoplasm.

To explore a possible link between seminal androgen concentrations and residual sperm cytoplasm, which constitutes one of the cytological anomalies of the spermatozoon middle piece, testosterone (T), delta4-androstenedione, the precursor of T during testicular androgen biosynthesis and the active metabolite of T, dihydrotestosterone, were assayed in the seminal fluid of 37 patients. A statistically significant correlation was found by linear regression (r = +0.380, P = 0.020, y = 0.02x + 0.45) between seminal T concentrations and the percentage of spermatozoa with a residual cytoplasmic droplet. Given the impact on fertility of residual sperm cytoplasm, assessment of seminal T concentrations could provide useful information on the fertility status of patients.

[Fibronectin and hyaluronic acid in seminal fluid: relationship with sperm count and percentage of typical forms]. / Fibronectine et acide hyaluronique dans le liquide séminal: relations avec la numération spermatique et le pourcentage des formes typiques.

Two components of the extra cellular matrix, hyaluronic acid (HA) and fibronectin (Fbn), were assessed in the seminal fluids of 29 patients submitted for diagnosis of infertility. The concentrations of HA and Fbn were elevated in seminal plasma in comparison to their ranges of concentration in normal sera. No correlations were found between seminal Fbn and sperm count per ml and/or ejaculate, normal sperm cytology and multiple anomaly index (MAI). The concentrations of HA were negatively correlated with sperm count per ml (p < 0.01, r = -0.5) and ejaculate (p < 0.005, r = -0.5), and also with sperm cytology (p < 0.001, r = -0.6). A positive correlation was found between seminal concentrations of HA and MAI (p < 0.001, r = +0.7). It seems that high seminal HA concentrations were in relation with the bad results of sperm count and sperm cytology. These data suggest that spermatozoon could be implicated, in part, in the seminal HA metabolism.

Relationship between seminal LDH-C4 and spermatozoa with acrosome anomalies.

Lactate dehydrogenase C4 (LDH-C4), the specific isozyme of LDH produced by germ cells, was assessed in the seminal plasma of 55 patients to test a potential link between LDH-C4 and the count per ml of spermatozoa with acrosome anomalies. A simple regression curve shows a statistically significant positive correlation between seminal LDH-C4 concentrations and count per ml of spermatozoa with acrosome anomalies (r = + 0.640, P < 0.0001). No correlation was found between seminal LDH-C4 concentrations and sperm motility. Though seminal LDH-C4 thus seems to be a poor indirect marker of energy produced by spermatozoa, in contrast, this LDH isozyme may be a useful indirect marker of acrosome anomalies.

Diagnostic accuracy of hyaluronan and type III procollagen amino-terminal peptide serum assays as markers of liver fibrosis in chronic viral hepatitis C evaluated by ROC curve analysis.

Diagnostic accuracy of two serum markers of liver fibrosis, hyaluronan (HA) and amino-terminal peptide of type III procollagen (P-III-P), was studied in a cohort of 326 untreated patients with chronic viral hepatitis C. Both P-III-P (RIA-gnost P-III-P, Behring Diagnostic) and HA (HA-test, Pharmacia) serum concentrations correlated with the histological grades of liver fibrosis (P < 0.001). Receiver-operating characteristic (ROC) curves showed that serum HA had greater diagnostic performance than P-III-P, both for discriminating patients with extensive liver fibrosis from those with no or mild fibrosis (area under the ROC curves: 0.864 vs 0.691, P <0.001) or for discriminating patients with cirrhosis from those without cirrhosis (area under the ROC curves: 0.924 vs 0.734, P <0.001). At cutoff values of 0.8 kU/L for serum P-III-P and 85 micrograms/L for serum HA, sensitivities were 70.0% and 64.5%, and specificities were 63.4% and 91.2%, respectively, for discriminating patients with extensive liver fibrosis from those with no or mild fibrosis. At the cutoff values of 1.0 kU/L for serum P-III-P and 110 micrograms/L for serum HA, sensitivities were 60.0% and 79.2%, and specificities were 74.0% and 89.4%, respectively, for discriminating patients with liver cirrhosis from those without cirrhosis. We conclude that, because the diagnostic accuracy of serum HA is greater than that of serum P-III-P as a marker of liver fibrosis, serum HA should be preferred when monitoring liver fibrosis in patients with chronic viral hepatitis C.

Plasma fibronectin kinetics during uncomplicated acute myocardial infarction.

Plasma fibronectin (Fbn) was assessed daily in 9 men admitted to the cardiology care unit for uncomplicated acute myocardial infarction (AMI). In a control group of twenty healthy men, plasma Fbn was 0.290 +/- 0.0417 g/l-1 (mean +/- SD). In 6 hypertensive AMI patients, Fbn levels were increased to a maximum of 0.461 +/- 0.0294 g/l-1 at day five (5.5 +/- 0.84 days) and returned to the range of the control group values 56 +/- 32.8 hours afterwards. Three normotensive patients had higher Fbn results without returning to the control group range at the end of the hospitalization (0.734 +/- 0.0209 gl-1). Plasma Fbn could in part be implicated in the repair process and/or in the limitation of the cardiac necrosis extension. Moreover, it seems that a link exists between plasma Fbn kinetics and hypertension and/or normotension during uncomplicated AMI.

Plasma fibronectin kinetics during uncomplicated acute myocardial infarction

Plasma fibronectin (Fbn) was assessed daily in 9 men admitted to the cardiology care unit for uncomplicated acute myocardial infarction (AMI). In a control group of twenty healthy men, plasma Fbn was 0.290ñ 0.0417g/l-1 (meanñSD). In 6 hypertensive AMI patients, Fbn levels were increased to a maximum of 0.461ñ0.0294 g/l-1 at day five (5.5 ñ 0.84 days) and returned to the range of the control group values 56 ñ 32.8 afterwards. Three normotensive patients had higher Fbn results without returning to the control group range at the end of the hospitalization (0.734 ñ 0.209 gl-1). Plasma Fbn could in part be implicated in the repair process and/or in the limitation of the cardiac necrosis extension. Moreover, it seems that a link exists between plasma Fbn kinetics and hypertension and/or normotension during uncomplicated AMI. (AU)

Plasma fibronectin kinetics during uncomplicated acute myocardial infarction

Plasma fibronectin (Fbn) was assessed daily in 9 men admitted to the cardiology care unit for uncomplicated acute myocardial infarction (AMI). In a control group of twenty healthy men, plasma Fbn was 0.290ñ 0.0417g/l-1 (meanñSD). In 6 hypertensive AMI patients, Fbn levels were increased to a maximum of 0.461ñ0.0294 g/l-1 at day five (5.5 ñ 0.84 days) and returned to the range of the control group values 56 ñ 32.8 afterwards. Three normotensive patients had higher Fbn results without returning to the control group range at the end of the hospitalization (0.734 ñ 0.209 gl-1). Plasma Fbn could in part be implicated in the repair process and/or in the limitation of the cardiac necrosis extension. Moreover, it seems that a link exists between plasma Fbn kinetics and hypertension and/or normotension during uncomplicated AMI.

Changes in systemic gonadal and adrenal steroids in asymptomatic human immunodeficiency virus-infected men: relationship with the CD4 cell counts.

Serum sex hormone-binding globulin (SHBG), testosterone, non-SHBG-bound testosterone, androstenedione, dihydrotestosterone (DHT), dehydroepiandrosterone (DHEA), follicle-stimulating hormone (FSH), luteinizing hormone (LH) and cortisol were measured in 58 homosexual men seropositive for human immunodeficiency virus (HIV), all clinically asymptomatic (Centers for Disease Control 1993 classification stage A). The HIV patients were divided into four groups according to the CD4 lymphocyte count--group 1 (more than 500/microliters, N = 14), group 2 (between 350 and 500/microliters, N = 16), group 3 (between 200 and 349/microliters, N = 22) and group 4 (less than 200/microliters, N = 6)--and compared with 11 antibody-negative men as controls. The SHBG levels were significantly increased in groups 1, 2, 3 (p < 0.01) and 4 (p < 0.05) compared with controls, with no differences between groups of patients. Compared with controls, testosterone concentrations were significantly lower in group 4 (p < 0.05) and non-SHBG-bound testosterone levels were significantly lower in groups 1 (p < 0.05), 2 (p < 0.01), 3 (p < 0.001) and group 4 (p < 0.001); DHT and androstenedione levels were significantly lower in group 4 (p < 0.05) and DHEA levels were significantly lower in group 2, group 3 (p < 0.01) and group 4 (p < 0.05) than in controls. Cortisol levels were significantly increased in groups 1 and 4 (p < 0.05) and FSH and LH concentrations were not significantly higher in HIV-infected men than in controls. Also, the DHEA, androstenedione, non-SHBG-bound testosterone and DHT levels were correlated with CD4 cell counts, showing that hypogonadism occurs as the CD4 lymphocytes decrease.(ABSTRACT TRUNCATED AT 250 WORDS)

The intestinal permeability test applied to the diagnosis of food allergy in paediatrics.

Intestinal permeability was evaluated in 17 healthy young children (control group), aged 4.3 +/- 1.9 years (mean +/- SD), weight 15.4 +/- 5.5 kg and in 11 infants with food allergy (allergic group, aged 2.3 +/- 1.6 years, weight 9.5 +/- 1.5 kg). In the two groups, the urinary excretion of two non-metabolized sugars (mannitol, lactulose) was measured by gas-chromatography after oral administration. Measurement of urinary content of these sugars 5 hours after administration in the control group showed a clearance of 20.2 +/- 8.6% for mannitol and 0.44 +/- 0.18% for lactulose. The lactulose/mannitol ratio in the same group was equal to 0.023 +/- 0.008. When compared to the control group, the 11 patients of the allergic group presented a normal mannitol urinary excretion (16.5 +/- 13.4%, p = NS, Student's t-test) and an increase in the lactulose excretion (1.36 +/- 0.92%, p < 0.001). Moreover, the allergic group showed a lactulose/mannitol ratio that was significantly different (0.105 +/- 0.071, p < 0.001). The intestinal permeability test for the diagnosis of food allergies seems to be a sensitive and non-invasive test that is well suited to the paediatric practice.

The intestinal permeability test applied to the diagnosis of food allergy in paediatrics

Intestinal permeability was evaluated in 17 healthy children (control group), aged 4.3 ñ 1.9 years (mean ñ SD), weight 15.4 ñ 5.5 kg and in 11 infants with food allergy (allergic group, aged 2.3 ñ 1.6 years, weight 9.5 ñ 1.5 kg). In the two groups, the urinary excretion of two non-metabolized sugars (mannitol, lactulose) was measured by gas-chromatography after oral administration. Measurement of urinary content of these sugars 5 hours after administration in the control group showed a clearance of 20.2 ñ 8.6 percent for mannitol and 0.44 ñ 0.81 percent for lactulose. The lactulose/mannitol ratio in the same group presented a normal mannitol urinary excretion (16.5 ñ 13.4 percent, p = NS, Student's t-test) and an increase in the lactulose excretion (1.36 ñ 0.92 percent, p<0.001). Moreover, the allergic group showed a lactulose/mannitol ratio that was significantly different (0.105 ñ 0.071,p<0.001). The intestinal permeability test for the diagnosis of food allergies seems to be sensitive and non-invasive test that is well suited to the paediatric practice (AU)

The intestinal permeability test applied to the diagnosis of food allergy in paediatrics

Intestinal permeability was evaluated in 17 healthy children (control group), aged 4.3 ñ 1.9 years (mean ñ SD), weight 15.4 ñ 5.5 kg and in 11 infants with food allergy (allergic group, aged 2.3 ñ 1.6 years, weight 9.5 ñ 1.5 kg). In the two groups, the urinary excretion of two non-metabolized sugars (mannitol, lactulose) was measured by gas-chromatography after oral administration. Measurement of urinary content of these sugars 5 hours after administration in the control group showed a clearance of 20.2 ñ 8.6//for mannitol and 0.44 ñ 0.81//for lactulose. The lactulose/mannitol ratio in the same group presented a normal mannitol urinary excretion (16.5 ñ 13.4//, p = NS, Student's t-test) and an increase in the lactulose excretion (1.36 ñ 0.92//, p<0.001). Moreover, the allergic group showed a lactulose/mannitol ratio that was significantly different (0.105 ñ 0.071,p<0.001). The intestinal permeability test for the diagnosis of food allergies seems to be sensitive and non-invasive test that is well suited to the paediatric practice

Immunoradiometric assay of human intact proinsulin applied to patients with type 2 diabetes, impaired glucose tolerance, and hyperandrogenism.

We describe an immunoradiometric assay for human intact proinsulin in serum. In this method, one monoclonal antibody, coated onto polyacrylamide beads, cross-reacts with proinsulins and insulin. A sandwich is formed with intact proinsulin, split (65-66) proinsulin, and des (64-65) proinsulin binding with an 125I-labeled monoclonal antibody specific for an epitope at the intact B-C junction of proinsulin. Because split (65-66) and des (64-65) proinsulin concentrations are very low in serum, this assay essentially measures intact proinsulin. When we used 1-mL serum samples, the mean detection limit was 0.4 pmol/L. Mean proinsulin concentrations (pmol/L) were 3.4 (range 1-9.1) in healthy fasting subjects, 28.5 (9.7-101) in patients with type 2 diabetes (treated with metformin and sulfonylureas), 5.0 (1.6-9.3) in women with hyperandrogenism and normal insulinemia, 10.3 (2.6-36) in women with hyperandrogenism and hyperinsulinemia, and 8.5 (4.8-21.3) in patients with impaired glucose tolerance.