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Biomedical subjects

F Dondero

Publications and source records attributed to F Dondero.

At least 37 records · Page 2Linked to original sources
PubMed2000

Azoospermia: incidence, and biochemical evaluation of seminal plasma by the differential pH method.

BACKGROUND: Recent artificial reproductive technologies (ART) have enabled even azoospermic subjects to take part in "in vitro fertilization" programs. The aim of the present study was: a) to evaluate the incidence of azoospermic male partners of infertile couples; b) to determine whether biochemical evaluation of such azoospermic males can help to discriminate between obstructive and secretory azoospermia. METHODS: Semen samples (n. 28,339; period 1990-1997) were analyzed according to WHO guidelines. In addition, in 108 azoospermic subjects, fructose, citric acid and L-carnitine (taken respectively as markers of seminal vesicle, prostate and epididymal function) were assayed using a new, faster technique, the differential pH method. RESULTS: The incidence of azoospermic semen samples was 4.76%. The biochemical analyses were indicative diagnostically, especially with regard to obstructive azoospermia. However, in secretory azoospermia, biochemical analysis of seminal plasma did not offer particularly helpful indications, except in cases of hypogonadism. CONCLUSIONS: Azoospermia accounts for a relatively high number of cases and justifies detailed attention to the diagnostic and therapeutic approach. Biochemical analysis of the seminal markers is a valid support for differential diagnosis between secretory obstructive azoospermia. It can therefore help in the correct recruitment of such patients for ART programs.

Biochemistry↗
PubMed1999

Immature germ cell separation using a modified discontinuous Percoll gradient technique in human semen.

The difficulty of identifying immature germ cells in unstained, fresh semen has led most laboratories to use the broad definition 'round cells' to indicate cells other than spermatozoa, thus grouping together both leukocytes and immature germ cells. This is also the case in research andrology, where very little attention has been given to immature germ cells in the semen apart from some rare exceptions, such as the attempts to study meiosis. Here we report on the use of a discontinuous Percoll gradient method modified to enable the best separation possible of immature germ cells from the other cells found in the ejaculate, in order to obtain a cellular suspension free of spermatozoa. Our technique (intra-assay variation in duplicates < 10%) demonstrated a high immature germ cell concentration in gradient fractions with 30% to 45% Percoll with a small contamination (1.5-6%) of leukocytes, confirmed by May-Grünwald-Giemsa staining, immunofluorescence and cytofluorimetry. The concentrations of immature germ cells ranged from zero in obstructive azoospermia to 2.0 x 10(6)/ml in oligozoospermia and genital tract infection. The purified immature germ cell suspensions obtained can be useful for diagnostic and research purposes.

Cell Separation↗
PubMed1999

[Intra-spermatic L-carnitine and survival of sperm motility].

BACKGROUND: The aim of this work was to evaluate the intrasperm carnitine (L-C) content related to sperm motility survival in bovine cervical mucus and in culture medium (Tyrode solution). METHODS: The following subjects were selected: 15 fertile normospermic subjects (according to WHO guidelines) and 31 male partners of infertile couples (semen profile: volume > 2.0 ml; concentration > 20 x 10(6)/ml, progressive motility > or = 25% (WHO categories "a" and "b") after fluidification, abnormal forms < 70%; round cells < 1.0 x 10(6)/ml). After standard semen analysis, the samples were subdivided into three aliquots in order to carry out: a) intrasperm L-C (free, total and acetylated) assay; b) sperm motility survival in bovine cervical mucus; c) sperm motility survival in culture medium. RESULTS: A strict correlation was found between L-C (total and acetylated) content and motility survival in cervical mucus. This is probably due to the fact that in cervical mucus lipids are an important energy source for sperm and to metabolize these lipids intrasperm L-C is essential. Therefore, L-C content can be considered as an indicator of sperm motility life-span in cervical mucus. A significant correlation, even if reduced compared with cervical mucus, was also observed between L-C (total and acetylated) content and sperm motility survival in the biological medium. This probably is because the L-C system modulates the reserves of free CoA, essential to the tricarboxylic acid cycle function. CONCLUSIONS: The intrasperm L-C deficit could be due to; a) alterations in the L-C uptake mechanisms in the epididimys due to inflammatory processes; b) lack of testosterone (L-C uptake is androgen-dependent). Therefore, the therapeutic implication of this finding is that where hypomotility is due to intrasperm L-C deficit, exogenous L-C administration or improvement of L-C epididymal testosterone-dependent uptake could promote the acquisition of sperm motility.

Animals↗
PubMed1999

Is hyperdiploidy of immature ejaculated germ cells predictive of testis malignancy? A comparative study in healthy normozoospermic, infertile, and testis tumor suffering subjects.

The possibility of diagnosing neoplastic testis pathologies by studying immature germ cells released from the seminiferous epithelium and present in the semen has been reported. It has been suggested that carcinoma in situ (CIS) of the testis and testis tumor may be identified by studying specific surface antigenic determinants or ploidy of chromosome 1 of malignant germ cells recovered from the semen. A noninvasive diagnostic approach of this type would be of great interest if we consider that CIS is supposed to precede the development of testicular germ cell tumors and that the frequency of that preneoplastic condition is increased in specific andrologic pathologies. To evaluate the reliability of this diagnostic approach, we have quantified the presence of immature hyperdiploid germ cells in the seminal fluid of normal healthy subjects, of infertile oligozoospermic patients affected by maldescended testis or left vancocele, and of patients suffering from CIS or testis tumor. Cell ploidy was evaluated on seminal cell fractions highly enriched in immature germ cells, by means of in situ hybridization with a DNA-probe specific for chromosome 1. Our observations indicate that chromosome 1 hyperdiploidy is not necessarily a predictive parameter of testis tumoral pathologies. The percentage of hyperdiploid immature seminal germ cells is in fact increased in nontumoral pathologies associated with infertility, such as cryptorchidism and left varicocele.

Adult↗
PubMed1999

Semen analysis and seminal plasma biochemistry in two brothers with familial reciprocal translocation 46,XY,t(8;9)(p10;p10) transmitted by the father.

Here we report familial balanced translocation (46,XY,t(8;9)(p10;p10) inherited by two brothers from the father. The first subject, 33 years old, was examined after 3 years of childless marriage. Semen analysis showed severe oligozoospermia, hypomotility and teratozoospermia. His brother, 29 years old, volunteered to join this study. He had not yet tried to start a family. Semen analysis showed dyspermia, but the condition was much less serious than his brother's. In both subjects, the secretory function of the epididymis and the genital tract accessory glands did not seem to be affected by this particular translocation. This case report highlights the importance of cytogenetic analysis in dealing with infertile patients suffering from severe dyspermia apparently sine causa. This is particularly the case when certain Assisted Reproductive Technology (ART) programs are recommended, where natural selection of the sperm no longer takes place (e.g. ICSI). The possible risk of an increase in fetal chromosomal abnormalities would suggest genetic counselling in all such situations.

Adult↗
PubMed1998

Sperm parameters after early left varicocele treatment.

OBJECTIVE: We evaluated whether early treatment for left varicocele can improve sperm parameters. DESIGN: Patient follow-up and comparison with control groups. SETTING: Surgical department of a Children's Hospital and a university infertility-care center. PATIENT(S): Nineteen patients who underwent surgery for left varicocele as adolescents (mean +/- SD, 14.11 +/- 1.59 years) were followed up as adults (mean +/- SD, 19.37 +/- 1.16 years). MAIN OUTCOME MEASURE(S): A testicular examination was performed and varicocele staging was done at the time of surgery. Andrologic examination and semen analysis were done 2-8 years later. Semen results were compared with those of two age-matched control groups of 19 healthy, randomly selected subjects and 19 unoperated patients suffering from left varicocele of the same degree. RESULT(S): The mean values of sperm parameters in the patient study group were similar to those of the healthy controls except that the sperm concentration was significantly lower. However, sperm motility and morphology mean values in the patient study group were significantly higher than those of the unoperated patients. CONCLUSION(S): This study indicates that surgery in adolescence can be useful, possibly because the testis is developing at that age. Large-scale, double-blind controlled trials on early treatment of left varicocele are needed to identify possible markers of disease severity.

Adolescent↗
PubMed1997

The application of the differential pH method to the biochemical evaluation of seminal plasma.

OBJECTIVE: To apply the differential pH method to the biochemical evaluation of seminal plasma. DESIGN AND METHODS: Fructose, citric acid and free L-carnitine concentration in human seminal plasma were measured by the differential pH method. These are routinely taken as markers of seminal vesicle, prostate and epididymal function, respectively. The pH differential method was then compared with standard spectrophotometry. RESULTS: The linearity, recovery and imprecision calculation for each assay were found to be reliable and the method correlated well with the reference spectrophotometric technique. In addition, the pH differential method showed certain advantages compared to spectrophotometry: a) speed of procedure (40 to 120 sec/assay); b) ability to measure the analytes even in small volumes of seminal plasma (10 microL to 50 microL) without deproteinization or dilution, thus eliminating manual operations. CONCLUSIONS: The differential pH method is an innovative approach to the biochemical assessment of seminal plasma.

Carnitine↗
PubMed1997

Unilateral cryptorchidism corrected in prepubertal age: evaluation of sperm parameters, hormones, and antisperm antibodies in adult age.

OBJECTIVE: To evaluate whether prepubertal orchidopexy for unilateral cryptorchidism can reduce adult dysspermia. DESIGN: Patient follow-up comparison with control groups. SETTING: Surgical and endocrinologic sections of a children's hospital and a university infertility-care center. PATIENT(S): Seventy-one patients with unilateral cryptorchidism who underwent orchidopexy in prepubertal age (6.4 +/- 2.8 years) were followed up as adults (20.0 +/- 2.8 years). MAIN OUTCOME MEASURE(S): Patients underwent testicular examination and hormonal evaluation, 49 of these had semen analysis and antisperm antibody tests. Semen results were compared with those of two age-matched control groups: a group of 20 healthy, randomly selected subjects and a group of 20 patients operated on in postpubertal age for cryptorchidism. RESULT(S): Unilateral reduced testis size was found in 30.1% of patients, eight patients had a low LH level, eight had a low T level, and none had abnormal FSH values. Antisperm antibodies were found in 1 of 49 cases. Cluster analysis of sperm parameters showed that the mean values of patients were worse than those of the healthy controls but better than those of the subjects operated on in postpubertal age. CONCLUSION(S): This study indicates that prepubertal orchidopexy can given better results than postpubertal correction.

Adult↗
PubMed1997

The in-vitro effects of nicotine and cotinine on sperm motility.

Swim-up spermatozoa from the seminal samples of non-smokers, usually not exposed to passive smoking, were treated in vitro with nicotine (NIC) and cotinine (COT) at the average levels found in smokers' seminal plasma and at levels 500 times higher than this average. This was done to evaluate the action of these drugs on sperm motility. Each sample was allowed to swim up in Tyrode's solution with or without the drug; the study was carried out at time 0 and +1, +2, +4, +8 and +24 h of incubation, using a light microscope and a CASA system (experiment 1). In addition, the direct action of smoke on spermatozoa was studied using aspirated cigarette smoke (experiment 2). Kinetic parameters were then measured at 30 min, 45 min and 60 min starting from the last smoke injection. The first experiment showed that NIC and COT at average levels did not produce statistically significant variations of the kinetic parameters studied up to 24 h. However, the much higher concentration significantly altered all the kinetic variables in relation to the time of incubation. The second experiment with smoke in toto demonstrated a sharp reduction in all the sperm kinetic parameters. This reduction was seen after 30 min exposure to smoke and increased progressively until almost complete immotility at 1 h of exposure. These results suggest that NIC and COT are not responsible for the harmful effects of cigarette smoke on sperm kinetic parameters reported in the literature.

Adult↗
PubMed1997

Antisperm antibody detection: 1. Methods and standard protocol.

PROBLEM: Today, antisperm antibody (ASA) detection is one of the most important steps in the evaluation of male infertility. This practice is generally accepted even though there is still some disagreement about the meaning of antisperm immunity and a good deal of controversy about the test regarded as the most suitable for the detection of antibodies directed against sperm antigens. International workshops have tried to standardize universally accepted protocols. A panel of three or four methods is generally advised to provide a correct and complete screening of patients with antisperm immunity. METHOD OF STUDY: This paper reports on the results of a serum exchange workshop for the standardization of the ASA detection in the sera carried out under the auspices of the Italian Society of Endocrinology, which was used as reference laboratory. A careful description of the most widely used methods is reported and proposed as a standard protocol also on the basis of the results of the correlation studies carried out by our group is also reported. CONCLUSIONS: If the proposed methods and procedures will be accepted by the members of the Alps-Adria Society for Immunology of Reproduction, a Study Group for Sperm Antibody Testing, could start its activity having the goal of reaching a consensus on methodology and carrying out a European serum exchange workshop.

Agglutination Tests↗
PubMed1997

Antisperm antibody detection: 2. Clinical, biological, and statistical correlation between methods.

PROBLEM: In clinical andrology, the detection of antisperm antibodies (ASA) is regarded as one of the most important steps in the study of male infertility. This practice is generally accepted even though there is still some disagreement about the true meaning of antisperm immunity, and there remains a good deal of controversy about the test regarded as the most suitable for the detection of antibodies directed against sperm antigens. International Workshops have tried to standardize universally accepted protocols. A panel of three or four methods is generally advised to provide a correct and complete screening of patients with antisperm immunity. The aim of the present paper is to report our studies on the correlation between direct methods (IBT, MAR test) and indirect methods (gelatin agglutination test [GAT], and tray agglutination test [TAT]) and to establish whether biological models can explain the antibody test results. An attempt was also made to establish a "predictive threshold" to explain even apparently discordant direct and indirect results.

Agglutination Tests↗
PubMed1997

[Seminal and biochemical features in a subject with male phenotype and 46, XX genotype].

Semen parameters and biochemical seminal markers in a 46,XX male subject were evaluated; Southern blot analysis with Y-DNA probes showed the absence of the SRY (Sex-determining Region of Y). The semen analysis, carried out three times, showed azoospermia, reduced volume (< 0.5 ml) and reduced pH value. Assays of the biochemical markers of the genital tract gave a profile very similar to that seen in cases of blockage of the ejaculatory ducts. This is probably due to inadequate differentiation of the genital tract adnexal glands leading to aplasia of the seminal vesicles. As a consequence, the ejaculate is formed of prostatic fluid only. The authors would underline the difficulties of counselling these subjects. This is particularly difficult where, as in this case, the genetic diagnosis is carried out in adulthood. In addition to the psychological stress of confirmed sterility, the patient must inevitably undergo the trauma of questioning his sexual identity.

Adult↗
PubMed1996

Effects of prolonged autovehicle driving on male reproduction function: a study among taxi drivers.

This study had the purpose of exploring the possible association between the work exposures of professional drivers and their reproductive health, by studying a group of 201 taxi drivers in the city of Rome. Data on work and reproductive history were collected by interviews. Biological markers examined in 72 subjects included salivary testosterone levels, sperm quality (i.e., sperm concentration, sperm morphology, and motility), and fertility experience, including time to pregnancy. Their spermatologic profile was compared with that of a control group of 50 healthy subjects of similar age and smoking habits. The results showed that taxi drivers, compared to the controls, had a significantly lower prevalence of normal sperm forms (45.8% vs. 64.0%); this was particularly true for those with a longer time on this job. This result was confirmed by a multivariate analysis in which confounders such as age, smoking, and alcohol consumption were controlled. The other sperm parameters did not differ in the study and the control groups. Among the life-style factors, we found smoking to be associated with poorer sperm morphology. Moderate alcohol consumption was associated with a better seminologic profile, while the pattern in respect to coffee intake was inconclusive. Subjects with poor semen quality also more frequently exhibited longer time to pregnancy of their partner. The results suggest that prolonged urban automobile driving might be a risk factors for sperm quality, and particularly for sperm morphology, but the finding needs further confirmation.

Adult↗
PubMed1996

Semen analysis in HIV seropositive men and in subjects at high risk for HIV infection.

The main purpose of this research was (i) to perform a comparative study of sperm parameters in human immunodeficiency virus (HIV) seropositive and high risk subjects in order to identify any possible alterations in the semen which specifically result from HIV infection and (ii) to study the p24 antigen as an early diagnostic marker of infection in high risk subjects. HIV seropositive subjects showed no significant variations regarding sperm densities, motility and viscosity compared to high risk subjects and controls. On the other hand, these HIV seropositive subjects showed (a) a significantly higher percentage of cytoplasmic droplet forms and immature germ cells, perhaps caused by an early failure of epididymal function and/or by a condition of stress affecting spermatogenesis after HIV infection and (b) a significantly higher level of spermiophage cells, suggesting that HIV activates mechanisms that increase spermiophagy. In addition, HIV seropositive men showed a significant positive correlation between blood CD4+ and sperm motility as well as a significant inverse correlation between CD4+ and sperm abnormalities. This is perhaps due to a decrease in testosteronaemia leading to defective epididymal sperm maturation. To date, p24 has not been found in the serum or seminal plasma of high risk subjects. The longitudinal study in progress should provide further information on this point.

Adult↗
PubMed1996

Gram staining applied to human spermatozoa: a simple method for studying chromatin condensation status.

Gram staining applied to human spermatozoa from fertile donors is described. The stain revealed populations of Gram-positive and Gram-negative spermatozoa. Data showed a significant and progressive decrease in the percentage of Gram-positive spermatozoa at different times during the chromatin decondensation procedure (SDS-BSA and SDS-EDTA). No significant correlation could be found between Gram staining and other functional tests used for spermatozoa; only the aniline blue staining test showed a poor correlation. Our study demonstrates that normal spermatozoa with regular chromatin condensation appear Gram-positive, while spermatozoa with altered chromatin condensation appear Gram-negative.

Cell Nucleus↗
PubMed1996

[Pregnancy in women undergoing a kidney transplant. Our experience and a review of the literature].

The chances of pregnancy for uremic women are usually very low, because of hormonal balance changes which determine a strong reduction in fertility. Epidemiological studies reveal that pregnancy in hemodialyzed women in fertile patients 4.6-6 months after a well functioning kidney transplant, one fertile transplanted woman over 50 can become pregnant. In the first transplant era, pregnancy after kidney transplant was considered "a big hazard", especially because of the possible side-effects of immunosuppression drugs on foetus development, and the risk of a worsening in the mother's renal function. Therefore, women were strongly recommended to avoid pregnancy. More recently, several reported papers have shown that pregnancy can be safely carried on also by transplanted women, under careful criteria and monitoring. Our experience too, even if limited in number (4 patients) reported in this article confirms this conviction.

Female↗
PubMed1995

Environmental tobacco smoke: nicotine and cotinine concentration in semen.

Forty-three nonsmokers attending an infertility clinic reported their degree of passive exposure to environmental tobacco smoke (ETS) and provided samples of blood and seminal plasma. Levels of nicotine and cotinine in biological fluid were evaluated by a radioimmunoassay. The data indicate that exposure to ETS results in measurable nicotine and cotinine levels in seminal plasma. Seminal plasma cotinine concentration also shows a significant positive correlation with degree of reported exposure.

Adult↗