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Biomedical subjects

C E Depuydt

Publications and source records attributed to C E Depuydt.

16 recordsLinked to original sources
PubMed2006

Improved endocervical sampling and HPV viral load detection by Cervex-Brush Combi.

OBJECTIVE: Liquid-based cytology (LBC) for cervical screening is becoming increasingly used. Together with SurePath LBC, various collecting devices can be utilized, among which the Cervex-Brush is the most widely used. The new Rovers Cervex-Brush Combi combines the advantages of the Cervex-Brush with the EndoCervex-Brush increasing sampling of the endocervical canal. The objective of this study was to analyse and to compare the Cervex-Brush Combi with the Cervex-Brush for the collection of squamous and endocervical cells, human papillomavirus (HPV) typing/quantification and disease detection in SurePath LBC. METHODS: Using either the Cervex-Brush or the Cervex-Brush Combi 100 consecutive SurePath LBC samples were collected using each brush type. All 200 slides were read by the FocalPoint and screened by guided screening using slide wizards. The viral load of HPV type 16 E7, 18 E7, 31 E6, 33 L1, 33 E6, 35 E4, 39 E7, 45 E7, 51 E6, 52 L1, 52 E7, 53 E6, 56 E7, 58 L1, 58 E6, 59 E7, 66 E6 and 68 E7 was determined using a TaqMan-based real-time quantitative PCR analysis. RESULTS: The mean number of sampled squamous cells did not differ between the two brush types (54 963 versus 54 595 cells). The use of the Cervex-Brush Combi, however, resulted in a two- to threefold increase in the number of sampled endocervical cells (P < 0.00001). Using the Cervex-Brush Combi slightly more lesions were detected (three versus two low-grade squamous intraepithelial lesions), and resulted in the detection of more atypical squamous cells of undetermined significance (six versus three). In the Cervex-Brush group, 60% (3/5) of abnormal smears were positive for oncogenic HPV types, whereas 66.7% (6/9) of abnormal smears in the Cervex-Brush Combi group tested positive. The median HPV viral load for samples taken with the Cervex-Brush Combi was 0.1825 copies/cell and was significantly higher than in samples taken with the Cervex-Brush (0.0042 copies/cell) (P = 0.02). CONCLUSION: Sampling with the Cervex-Brush Combi resulted in the collection of the same amount of squamous cells, but in a two to threefold harvest of endocervical cells. This led to the detection of a higher viral load for oncogenic HPV and an increase in the number of detected abnormal smears.

Cohort Studies↗
PubMed2003

Thin-layer liquid-based cervical cytology and PCR for detecting and typing human papillomavirus DNA in Flemish women.

The objective of this study was to document the occurrence and to correlate the prevalence of different human papillomavirus (HPV) types with the cytological results on simultaneously performed thin-layer preparations in a large population of Flemish women. During 1 year, 69 290 thin-layer preparations were interpreted using the Bethesda classification system. Using an algorithm for HPV testing based on consensus primers and type-specific PCRs in combination with liquid-based cytology, we determined the occurrence and distribution of 14 different oncogenic HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68). Reflex HPV testing was performed on cytologically abnormal samples and on an age matched randomly selected control group with normal cervical cytology (n=1351). Correlation between cytology, age and prevalence for the 14 different high-risk HPV types is given. There is a significant increase in predominance of high-risk HPV types, with increasing abnormal cytology. Coinfection with multiple HPV types also increased with cytological abnormalities, and was highest in HSIL (16.7%). In Flanders, HSIL was most often associated with HPV types 16, 33, 35, 31, 18 and 51. Using thin-layer liquid-based cytology and PCR to detect HPV, it is feasible to screen large numbers of women.

Adolescent↗
PubMed2003

Ki-67 immunocytochemistry in liquid based cervical cytology: useful as an adjunctive tool?

AIMS: To test the ability of Ki-67 to detect cytological lesions in a screening setting and its use as a surrogate marker of human papillomavirus (HPV) infection. METHODS: A study of liquid based cytology, HPV DNA testing by MY09/MY11 consensus polymerase chain reaction (PCR), type specific PCRs, and Ki-67 immunocytochemistry on a randomly selected series of 147 patients. RESULTS: Comparison of the number of Ki-67 immunoreactive cells/1000 cells in the different cytological groups showed that the HSIL group yielded a significantly higher mean count than did the other groups. The number of Ki-67 immunoreactive cells/1000 cells was significantly higher in HPV-16 positive samples than in samples containing infections with other high risk types. Receiver operating characteristic curves indicated a test accuracy (area under curve) of 0.68, 0.72, and 0.86 for atypical squamous cells of undetermined significance (ASCUS), low grade squamous intraepithelial lesions (LSIL), and high grade squamous intraepithelial lesions (HSIL), respectively. Thresholds for 95% sensitivity were 0.07, 0.08, and 0.15 Ki-67 immunopositive cells/1000 cells for ASCUS, LSIL and HSIL, respectively. The threshold for 95% specificity was 1.9 Ki-67 immunopositive cells/1000 cells. CONCLUSIONS: Ki-67 immunocytochemistry can be applied to liquid based cytology. The accuracy and diagnostic indices of the test are good when compared with those of other techniques. As part of a panel of screening procedures, it could be used as an adjunct to liquid based cytology to identify HSIL, and as a surrogate marker of HPV-16 infection.

Biomarkers↗
PubMed2000

The effects of combined conventional treatment, oral antioxidants and essential fatty acids on sperm biology in subfertile men.

We evaluated the effects of combined conventional treatment, oral antioxidants (N-acetyl-cysteine or vitamins A plus E) and essential fatty acids (FA) on sperm biology in an open prospective study including 27 infertile men. The evaluation included sperm characteristics, seminal reactive oxygen species (ROS), FA of sperm membrane phospholipids, sperm oxidized DNA (8-OH-dG), and induced acrosome reaction (AR). Treatment did not improve sperm motility and morphology, nor decrease the concentration of round cells and white blood cells in semen. Sperm concentration increased in oligozoospermic men (7.4+/-1.3 to 12.5+/-1.9 million/ml). Treatment significantly reduced ROS (mean+/-SEM) (775.3+/-372.2 to 150.3+/-105.2 x 10(3)counts/10 second) and 8-OH-dG (45.3+/-10.4 to 16. 8+/-3.3 fmol/microg DNA). Treatment increased the AR (55.1+/-2.2 to 71.6+/-2.2%), the proportion of polyunsaturated FA of the phospholipids, and sperm membrane fluidity. The overall pregnancy rate was 4.5% in 134 months. The per month pregnancy rate tended to be higher in partners of (ex)-smokers (7.15%, n=14,70 months) than in never-smokers (1.6%, n=13,64 months) (OR:4.57, 95% Cl:0.55-38.1).

Acetylcysteine↗
PubMed1998

The fatty acid composition of phospholipids of spermatozoa from infertile patients.

The lipid composition of the sperm membrane has a significant effect upon the functional characteristics of spermatozoa. In the present study we investigated the fatty acid (FA) composition of subpopulations of spermatozoa separated on a discontinuous Percoll gradient (47:90%) and the FA composition of phospholipids (PL) of sperm heads and tails in both normal and abnormal semen samples. In normozoospermic samples, polyunsaturated fatty acids (PUFA) represented 34.0 +/- 1.3 (mean +/- SE, mole %) and 25.6 +/- 1.2% of total FA of PL of the 47 and 90% Percoll fractions respectively. Docosahexaenoic acid (22:6omega3, DHA) contributed to more than 60% of total PUFA. DHA was significantly lower in both the 47% (P < 0.05) and the 90% (P < 0.01) Percoll fractions of oligozoospermic samples and in the 90% Percoll layer of asthenozoospermic samples (P < 0.01), compared with normozoospermic samples. The omega6/omega3 ratio was significantly increased in both Percoll fractions of samples with oligozoospermia (47%, P < 0.001 and 90%, P < 0.001) or with asthenozoospermia (47%, P < 0.05 and 90%, P < 0.001) compared with normozoospermic samples. The oxidative potential index (OPI) of spermatozoa recovered from the 47% Percoll layer was significantly higher (P < 0.0001) than of those recovered from the 90% Percoll. Mean melting point (MMP), an index of membrane fluidity, was significantly lower in head than in tails (P < 0.01) of spermatozoa, and also in both the 47% (P < 0.01) and 90% (P < 0.001) Percoll fractions of normozoospermic samples in comparison with oligozoospermic samples. The MMP was significantly higher (P < 0.05) in samples of patients with idiopathic oligo/asthenozoospermia, varicocele, and male accessory gland infection (MAGI). These differences in FA composition of PL in subpopulations of human spermatozoa, and in their heads and tails may be related to sperm maturity and to differences in physiological function.

Epididymitis↗
PubMed1998

White blood cells cause oxidative damage to the fatty acid composition of phospholipids of human spermatozoa.

The lipid composition of the sperm membrane has been shown to exert a significant effect upon the functional quality of spermatozoa. We have studied the effect of induced peroxidation and of the presence of polymorphonuclear white blood cells (WBCs) on the fatty acid composition of the phospholipids of human spermatozoa. The spermatozoa were fractionated by a discontinuous Percoll gradient in two fractions (47% and 90% Percoll). Induced peroxidation of spermatozoa was assessed by determining the production of thiobarbituric acid reactive substances (TBARS), mostly malondialdehyde, after incubation with ferrous sulphate and sodium ascorbate as a promoter of peroxidation. TBARS production after induction of peroxidation was correlated with the abundance of polyunsaturated fatty acids (PUFA)(r = 0.68, p < 0.0001), with the double bond index (r = 0.72, p < 0.0001), and with the oxidative potential index (r = 0.73, p < 0.0001) of fatty acids of phospholipids. In comparison with samples containing > 1 x 10(6) WBCs/mL, those with < 1 x 10(6) WBCs/mL contained higher proportions of PUFA (90% Percoll, p < 0.05; 47% Percoll, p < 0.05), total omega 3 fatty acids (90% Percoll, p < 0.05; 47% Percoll, p < 0.001), docosahexaenoic acid (90% Percoll p < 0.05; 47% Percoll, p < 0.05), and double bond index (90% Percoll, p < 0.05; 47% Percoll, p < 0.001). In addition, mean melting point was significantly lower (90% Percoll, p < 0.05; 47% Percoll, p < 0.001) in samples with < 1 x 10(6) WBCs, indicating higher membrane fluidity. The increase of TBARS production by spermatozoa after incubation with the xanthine-xanthine oxidase system and/or ferrous sulphate as promoter of peroxidation was associated with a significant decrease of PUFA. Incubation of spermatozoa with WBCs, with or without activation by phorbol ester, decreased the PUFA (p < 0.05). Also, TBARS production was increased (p < 0.01) after activation of WBCs with phorbol ester. Our data provide evidence that oxidative stress induced by WBCs has a damaging effect on the polyunsaturated fatty acids of sperm phospholipids which may result, amongst other effects, in decreased membrane fluidity.

Fatty Acids↗
PubMed1997

Purification and characterization of hepatocyte growth factor (HGF) from human seminal plasma.

Naturally occurring forms of hepatocyte growth factor/scatter factor (HGF/SF) have been purified by heparin-Sepharose chromatography, followed by cation exchange chromatography from a pool of human seminal plasma. Using an enzyme-linked immunosorbant assay, MDCK scatter assay, and Western blot analysis, it was found that, after heparin-Sepharose chromatography, human HGF/SF present in seminal plasma eluted in two different fractions, between 0.72 and 0.85 M NaCl (fraction I) and between 0.95 and 1.10 M NaCl (fraction II). Further purification of fraction I by cation exchange chromatography resulted again in two fractions which eluted at 0.2-0.4 and at 0.6-0.8 M NaCl. The fraction which eluted at 0.2-0.4 M NaCl consisted of two biologically less active heavy chains of the heterodimeric form of HGF/SF (107.1 U/ng immunoreactive HGF), with approximate molecular weights of 65 and 62 kDa under reducing conditions. The second fraction, which eluted at 0.6-0.8 M NaCl, revealed three bands with molecular weights of 87, 65 and 62 kDa, respectively. The 87 kDa form is thought to be a single chain precursor of HGF/SF devoid of biological activity. After subjecting fraction II to cation exchange chromatography, only one major peak eluted between 0.9 and 1.0 M NaCl, and consisted of two biologically active heavy chains of the heterodimeric form of HGF/SF (708.3 U/ng immunoreactive HGF), with approximate molecular weights of 65 and 62 kDa under reducing conditions. Nonreducing conditions for both fraction I and fraction II revealed only one band with a molecular weight of 68 kDa. The ratio ofpro-HGF/SF and less biologically active HGF/ SF (fraction I) over mature heterodimeric HGF/SF (fraction II) was approximately 1:3, in seminal plasma from sperm donors. In seminal plasma, pro-HGF/SF represents an 87 kDa glycoprotein which, apparently, is converted by limited proteolysis into several bands with molecular weights of 65 and 62 kDa. This is the first report showing the presence of pro-HGF/SF and heterodimeric mature HGF/SF, as well as less biologically active forms of HGF/SF in human seminal plasma.

Adult↗
PubMed1996

The receptor encoded by the human C-MET oncogene is expressed in testicular tissue and on human spermatozoa.

Because of its distinctive ability to act as a mitogen, a mitogen and a morphogen, hepatocyte growth factor/scatter factor (HGF/SF) has all the characteristics of a molecule able to function in regulatory networks of motility, such as the spermatogenic epithelium, and this through binding of its receptor p190MET (C-MET). In this study we report the expression of C-MET in the human seminiferous epithelium and on spermatozoa from men being treated for infertility and sperm donors. The presence of C-MET was demonstrated by immunochemistry on the cell membrane of spermatogonia, spermatocytes, spermatids and on spermatozoa, whereas Sertoli cells and Leydig cells did not show expression. Comparison of C-MET expression on spermatozoa of the 90% Percoll layer of subfertile patients and donors revealed clearly two distinct groups (unpaired t-test, P < 0.001), whereas comparison of C-MET expression on spermatozoa in the 47% Percoll layer was not significantly different between patients and donors. In addition, there was a significant inverse correlation between sperm concentration and the C-MET expression of spermatozoa in the 90% Percoll layer (r = -0.80, 95% confidence interval, -0.92 to -0.55; P < 0.0001), but not with the C-MET expression of spermatozoa in the 47% Percoll layer. In conclusion, the presence of C-MET was demonstrated in the seminiferous epithelium and on mature and immature spermatozoa, indicating a role for this growth factor receptor in the differentiation and/or migration that occurs during human spermatogenesis.

Blotting, Western↗
PubMed1995

MedCalc: a new computer program for medical statistics.

In recent years, the use and abuse of statistics in the medical literature has extensively been reviewed. Amongst others, the importance of the P-value has been challenged and the use of misleading graphics, including 3-dimensional displays, has been criticized. The ease of access to more complex statistical procedures, since the introduction of several statistical software packages for personal computers, has been identified as one of the factors involved in the misuse of statistics. Therefore, we have developed a new computer program that includes those statistical procedures commonly encountered in the medical literature and in statistical textbooks for medical researchers. More complex statistical analyses are not implemented in the software. If researchers with limited statistical training require more sophisticated statistical analyses, they should refer to a statistician, not to a more complete statistical software package.

Computer Graphics↗
PubMedDate not supplied

The clinical and biologic significance of serum inhibins in subfertile men.

Inhibin B is a marker of spermatogenesis and Sertoli cell function. The objective of this study was to evaluate the biologic significance of inhibins in subfertile men and the usefulness of inhibin B for the detection of male reproductive dysfunction. Forty-seven subfertile men were evaluated by semen analysis and clinical examination. In addition to semen analysis and hormone determinations, inhibins A and B (Serotec) in all 47 and inhibin A in 25 of these samples using another kit (Biosource) were measured. Higher inhibin B (median, range: 160.3, 81.8-328.5 pg/mL vs. 94.9, 15.6-389.7 pg/mL, P = 0.024) and lower FSH (P = 0.001) were detected in men with sperm concentrations > or =20 million/mL (n = 9), compared to oligozoospermia (sperm concentration <20 million/mL, n = 38). Inhibin B correlated significantly negatively with FSH, LH, and E2, and patient's age and positively with sperm concentration, testicular volume, and TSH. Multiple regression analysis indicated FSH, LH, E2, TSH, and age as the independent variables for inhibin B with a coefficient of determination (R) of 0.53. Simultaneous measurement of both FSH and inhibin B identified more cases with oligozoospermia than either hormone alone. Taking into account the body mass index, the age of the patient, and the indirect mixed antiglobin reaction (MAR) test result in addition to FSH and inhibin B led to the correct semen classification in 45 out of 47 cases. The simultaneous measurement of FSH and inhibin B, taking into account age, body mass index, and the indirect MAR test result appears accurate in identifying subfertility. Inhibin A is detectable in some subfertile men but its significance is not clear.

Adult↗
PubMedDate not supplied

Mechanisms and effects of male genital tract infection on sperm quality and fertilizing potential: the andrologist's viewpoint.

There are several mechanisms acting in synergism that can impair sperm characteristics of patients with accessory gland infection. In some cases, conventional sperm variables are disturbed with oligo and/or asthenozoospermia. In other patients, these sperm variables may appear normal, but the functional capacity of spermatozoa may be impaired. In particular, changes in the composition of the sperm membrane may result in reduced acrosome reactivity and capacity to fuse with the oolemma, and oxidative damage of the sperm DNA may induce mutagenesis. Changes in the biochemical make-up of seminal plasma can also reduce the in-vivo fertilizing capacity of spermatozoa, and infection-related disruption of the blood-testis barrier can induce the generation of anti-sperm antibodies and immunological infertility. Many of these functional abnormalities will not become evident upon 'basic semen analysis', which explains why some authors are unable to link infection of the accessory sex glands to subfertility. Also, functional and anatomical damage acquired as a result of infection is often permanent and not reversible by (antibiotic) treatment. Clearly, there are many more aspects of male accessory gland infection that require investigation. Available data should stimulate clinicians to place more emphasis on the prevention of infection-related infertility than on its treatment, as the latter is often unsuccessful.

Cytokines↗
PubMedDate not supplied

Cervical cytological screening and human papillomavirus DNA testing in Flanders.

The causal relationship between genital human papillomavirus (HPV) infection and cervical dysplasia/carcinoma has been recognised for some time. The aim of this study was to document the occurrence and distribution of HPV infection in the five provinces of the Flemish region in Belgium and to correlate the HPV DNA test results with the cytological results on simultaneously performed thin layer preparations of cervical cells. Out of a total screened group of 105107 samples, 1978 samples with cytological abnormalities were tested for HPV DNA using the MY09/MY11 consensus PCR. The mean age of the whole group was 36.9 years. The LSIL group, with a mean age of 33.6 years, was significantly younger than the other groups. There was no significant difference in HPV prevalence among the provinces. In four out of five provinces the HPV prevalence reached 100% in high-grade lesions. There is a significant increase in predominance of high-risk HPV types, with increasing abnormal cytology (17.9% WNL < 51.1% ASCUS < 83.8% LSIL < 97.2% HSIL). Three peaks of HPV DNA positivity were observed, a first at 22 yrs (82%), a second at 47 yrs (60%) and a third in women older than 65 yrs (52%). These results shed more light on HPV prevalence in Flanders and show that the MY09/MY11 consensus primer based detection system is very suitable for the detection of HPV infections in Flanders.

Adult↗
PubMedDate not supplied

Hormonal regulation of inhibin B secretion by immature rat sertoli cells in vitro: possible use as a bioassay for estrogen detection.

The influences of follicle-stimulating hormone (FSH), gonadal steroids, and culture time were studied in relation to inhibin B production by Sertoli cells of immature rats cultured in vitro. Sertoli cell-enriched cultures were established from 18-day-old rats and were maintained in medium supplemented with insulin, transferrin, and epidermal growth factor at 34 degrees C. A recently developed ELISA for the measurement of inhibin B was used to assess the effects of recombinant human FSH (rh FSH), testosterone (T), and estradiol (E2) on inhibin B production and accumulation in the culture media of Sertoli cell-enriched cultures and to optimize the cell culture system to serve as a bioassay for the detection and quantification of estrogens and estrogenlike substances. Prolonging the incubation time (24, 48, or 72 hours) of Sertoli cells with control medium without rh FSH, T, or E2 resulted in a time-dependent increase of inhibin B production. Incubation with rh FSH (1, 2.5, 5, or 10 U/L) caused a dose- and time-dependent increase of inhibin B production by Sertoli cells (but not by cultured Leydig cells), reaching a plateau at 5 U/L rh FSH. Addition of T in concentrations of 2.88, 5, or 50 ng/ml to medium without rh FSH and E2 significantly lowered the daily production rate of inhibin B (P < 0.05). In contrast, addition of E2 (0.01 and 0.1 ng/ml) caused a dose-responsive increase in inhibin B production after 24 and 48 hours. The relative increment of inhibin B production induced by E2 was maximal after 24 hours in the presence of 2.5 U/L rh FSH (acting synergistically) and in the absence of T. When these conditions are implemented, the Sertoli cell culture system may serve as a bioassay for estrogenic substances, and it may reflect the possibly harmful effect they may have on spermatogenesis.

Animals↗
PubMedDate not supplied

The relation between reactive oxygen species and cytokines in andrological patients with or without male accessory gland infection.

The presence of various cytokines, namely hepatocyte growth factor (HGF), interleukin-1 receptor antagonist (IL-1 RA), and interleukins (IL-1 alpha, IL-6, and IL-8), as well as the production of reactive oxygen species (ROS) was investigated in seminal plasma of fertile and infertile patients in order to evaluate the possible value of measuring these substances for the diagnosis of male accessory gland infection, and to assess the possible relationship between oxidative stress and cytokines during leucocytospermia and male accessory gland infection (MAGI). Our findings indicate that all of the measured cytokines seem to be produced locally as well as by white blood cells (WBC) and that, due to the presence of higher numbers of WBC, accessory gland infection may exert a deleterious effect on sperm quality through the production of ROS and/or of particular cytokines such as IL-1 alpha, IL-1 RA, and IL-8. The most specific marker for a sensitivity of 95% in discriminating between cases with or without MAGI is the measurement of IL-6 in seminal plasma. In the absence of WBC several cytokines are constitutively produced and correlate with sperm concentration (HGF, IL-8), alpha-glucosidase (IL-6), and gamma-glutamyltransferase activity (HGF). The measurement of these cytokines in semen may provide clinically useful information for the diagnosis of male accessory gland infection, as well as in the absence of WBC where it can provide information about certain mechanisms of male reproductive function and dysfunction.

Androgens↗
PubMedDate not supplied

Levels of hepatocyte growth factor/scatter factor (HGF/SF) in seminal plasma of patients with andrological diseases.

Hepatocyte growth factor/scatter factor (HGF/SF) has all the characteristics of a molecule suitable for functioning in regulatory networks of motility, such as the spermatogenic epithelium, where spermatogenic cells must migrate between the cells of Sertoli, and it exerts its effect through binding of its high-affinity receptor (c-met). Considering the findings that c-met receptor is expressed in the human testis and on spermatozoa, and that HGF/SF in seminal plasma consists of pro-HGF/SF, mature alphabeta-HGF/SF, and less active forms of HGF/SF, we investigated the concentration and biological activity of HGF/SF in seminal plasma and their correlation with parameters of spermatogenesis to obtain better insight into mechanisms that may be involved in the pathogenesis of male infertility. We also evaluated the potential value of assessment of hepatocyte growth factor concentration and its bioactivity for the diagnosis of certain pathological conditions of male reproduction. We studied the concentration and biological activity of HGF/SF in seminal plasma of normal men and of patients with a range of andrological diseases or conditions by measuring HGF/SF in seminal plasma by enzyme-linked immunosorbent assay and by scatter assay using Madin-Darby canine kidney epithelial cells. We identified three sources of HGF/SF in seminal plasma. In samples from vasectomized men (n = 30; 2.01 ng/ml) and in split ejaculate samples (n = 6; 1e fraction 2.75 ng/ml, 2e fraction 1.62 ng/ml), a prostatic origin can be certified. This HGF/SF has low biological activity (133.3 U/ml). In inflammation of the accessory sex glands (n = 40), a high amount of HGF/SF (3.04 ng/ml) can be generated by white blood cells and has moderate scatter activity (426.7 U/ml). In normozoospermic samples, there is a lower amount of HGF/SF (1.12 ng/ml), with strong scatter activity (1280.0 U/ml). Finally, the clear difference between the low amount of HGF/SF (1.06 ng/ml) with poor scatter activity (106.6 U/ml) in oligozoospermic samples (n = 28) and the high amount of HGF/SF (3.35 ng/ml) with strong scatter activity (853.3 U/ml) in samples from men with azoospermia of primary testicular failure (n = 18) suggests a mainly testicular origin, with different activity in different pathological conditions.

Animals↗