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Biomedical subjects

F Mazzilli

Publications and source records attributed to F Mazzilli.

At least 19 recordsLinked to original sources
PubMed2002

Simultaneous cycle sequencing assessment of (TG)m and Tn tract length in CFTR gene.

The lengths of the dinucleotide (TG)m and mononucleotide Tn repeats, both located at the intron 8/exon 9 splice acceptor site of the cystic fibrosis transmembrane conductance regulator (CFTR) gene whose mutations cause cysticfibrosis (CF), have been shown to influence the skipping of exon 9 in CFTR mRNA. This exon 9-skipped mRNA encodes a nonfunctional protein and is associated with various clinical manifestations in CF As a result of growing interest in these repeats, several assessment methods have been developed, most of which are, however, cumbersome, multi-step, and time consuming. Here, we describe a rapid methodfor the simultaneous assessment of the lengths of both (TG)m and Tn repeats, based on a nonradioactive cycle sequencing procedure that can be performed even without DNA extraction. This method determines the lengths of the (TG)m and Tn tracts of both alleles, which in our samples ranged from TG8 to TG12 in the presence of T5, T7, and T9 alleles, and also fully assesses the aplotypes. In addition, the repeats in the majority of these samples can be assessed by single-strand sequencing, with no need to sequence the other strand, thereby saving a considerable amount of time and effort.

Alleles↗
PubMed2001

A new objective method for scoring human sperm hyperactivation based on head axis angle deviation.

The aims of the present study were, first to develop a new method for evaluating sperm hyperactivation (HA) based on sperm head axis angle deviation, and second to apply this method in scoring selected sperm populations during capacitation. This was made possible by improving our original superimposed image analysis system (SIAS). The new option on the system enables us to determine the real angular deviation of the entire head in successive superimposed frames. The entire procedure for each spermatozoon requires 10-20 sec, according to the frame/rate utilized. A clear cut-off between the values of hyperactivated, transitional and non-hyperactivated spermatozoa was found at 11 and 21 frames/sec. However, at 6 frames/sec, a partial overlap between the three classes was found. We defined sperm activity as follows. At 11 frames/sec, hyperactivated: angle sum > or = 405 degrees; transitional: 200 degrees < or = angle sum < 405 degrees. At 21 frames/sec, hyperactivated: angle sum > or = 760 degrees; transitional: 350 degrees < or = angle sum < 760 degrees. The results, in agreement with previous research, show the peaks of activity at 3 h, dropping to a minimum at 6 h. This phenomenon was significantly more evident in fertile compared with subfertile semen samples.

Humans↗
PubMed2000

Effects of sildenafil (Viagra) administration on seminal parameters and post-ejaculatory refractory time in normal males.

Sildenafil is a specific inhibitor of phosphodiesterase (PDE) type 5 and represents a powerful therapy for male erectile dysfunction (ED) of different aetiology. Recently, sildenafil has been shown to restore erections in temporary ED related to the need of semen collection for assisted reproductive techniques. In this study, we investigated whether sildenafil administration modifies seminal parameters and/or erectile function in normal healthy volunteers. In a double-blind, randomized, placebo-controlled, cross-over two period investigation we enrolled 20 healthy male volunteers (mean +/- SE age 32 +/- 0.5 years). Subjects were not using any medication for the 3 month period prior to the study and were engaged in a stable relationship with proven fertility. The effects of sildenafil (100 mg) on seminal parameters and erectile function after audiovisual sexual stimulation were evaluated by semen analysis and by colour-Duplex ultrasound (the Resistive Index) respectively. In all subjects, sildenafil caused no changes in seminal and erection parameters when compared to placebo. Interestingly, sildenafil administration led to a marked reduction of the post-ejaculatory refractory time (10.8 +/- 0.9 min versus 2.6 +/- 0.7 min for placebo and sildenafil respectively; P < 0.0001). These results indicate that in normal subjects acute sildenafil treatment does not modify semen characteristics and has a positive influence over the resumption of erections following ejaculation in the presence of a continuous erotic stimulus.

3',5'-Cyclic-GMP Phosphodiesterases↗
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

Application of the upgraded image superimposition system (SIAS) to the assessment of sperm kinematics.

Dedicated software for semiautomatic objective motility assessment, based on image superimposition (SIAS), was upgraded in order to allow automatic reading of sperm tracks whilst keeping the advantages of visual check instead of the digital images of classic computer-aided sperm analysis (CASA) systems. The new system is very fast and accurate and allows an excellent reading of sperm motility in standard semen analysis. Moreover, using this system, an attempt was made to define numerically straight-line velocity (VSL), curvilinear velocity (VCL) and linearity (LIN) in each WHO sperm motility class. At 21 frames sec-1, ROC curves analysis determined the following: Class 1 (typical of WHO grade 'a'): VSL > or = 23 microns sec-1 and LIN > or = 0.58; class 2 (typical of WHO grade 'b'): VSL > 10 and < 23 microns sec-1 and LIN > or = 0.58; class 3: VSL > 10 microns sec-1 and LIN < 0.58 (this additional class was added to differentiate nonstraight progressive motility from classes 1 and 2); class 4 (typical of WHO grade 'c'): VSL < or = 10 microns sec-1. The numerical definition of sperm motility classes may contribute towards standardization in the objective evaluation of sperm kinematics.

Humans↗
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

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↗
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

[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

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↗
PubMed1995

Superimposed Image Analysis System (SIAS) software: a new approach to sperm motility assessment.

OBJECTIVE: To apply Superimposed Image Analysis System (SIAS) software (Delta Sistemi, Rome, Italy) to the objective analysis of sperm motility. DESIGN: To test a new procedure for sperm motility assessment based on image superimposition. RESULTS: The system can superimpose six sequential frames onto a monitor producing a final image with a motion effect similar to the multiple exposure photography method, but bypassing the film processing stage. Viewing the final image of a complete series of six superimposed frames allows the evaluation of the percentage of motile spermatozoa and their kinetic characteristics. Compared with multiple exposure photography, SIAS has the following main advantages: [1] the procedure is quick; [2] projecting the first frame alone allows the easy evaluation of sperm concentration; and [3] the system allows quick and easy analysis and measurement of each sperm trajectory frame by frame, thus avoiding misleading interpretations because of crossing sperm trajectories, collisions, and overlapping of motile spermatoza with round cells or immotile spermatozoa. CONCLUSIONS: Superimposed image analysis is a new approach to sperm motility assessment. It seems to be useful particularly in selected situations, such as oligozoospermia or in cases of increased round cell concentration. In these situations, SIAS can even serve as a support or an alternative to the computer-aided sperm analysis system widely used for objective sperm analysis.

Humans↗
PubMed1994

Superoxide anion in human semen related to seminal parameters and clinical aspects.

OBJECTIVE: To study the possible presence of extracellular superoxide anion (O2-) in untreated semen of normal and pathological subjects and to determine the possible relationship between superoxide anion levels, number of seminal leukocytes (WBCs) and seminal parameters. PATIENTS: Semen samples were analyzed from fertile normospermic subjects (n = 20) and patients (n = 132) attending the Outpatient Department of the 5th Medical Clinic. MAIN OUTCOME MEASURE: We evaluated superoxide anion levels by monitoring the reduction of cytochrome c. RESULTS: O2- was detected in 55.0% of fertile normospermic subjects. Both the percentage of subjects in whom O2- was detected and the mean values +/- SD of O2- were significantly higher in the male partners of infertile couples, in subjects with varicocele, and in subjects with positive sperm culture than normospermic subjects. No variations were seen in vasectomized subjects or in patients with past cryptorchidism. A high correlation was observed between O2- levels and WBCs (rs = 0.58), nonrapid, immotile sperm (rs = 0.64), percentage decrease of motility within a specified time frame (2 to 6 hours) (rs = 0.73), and sperm abnormalities (rs = 0.49). CONCLUSIONS: Superoxide anion in untreated semen is produced by WBCs and atypical and nonrapid, immotile sperm. A significant increase of superoxide anion was detected in the male partners of infertile couples and in some selected andrological conditions. The toxic effect of superoxide anion on sperm cells is dependent on the time of exposure and superoxide anion concentration.

Adult↗
PubMed1979

Treatment and follow-up of patients with infertility due to spermagglutinins.

A therapeutic trial based on pharmacologically induced azoospermia followed by the administration of corticosteroids was carried out in 48 patients with infertility due to spermagglutinins. Three types of responses were observed: type A, no modifications in the blood and spermagglutinating (SA) titer (19 cases); type B, disappearance or significant decrease in SA titer during treatment, with return to normal values upon resumption of spermatogenesis (7 cases); and type C, disappearance or significant decrease in SA titer for at least 1 year after stopping treatment (22 cases). Within the 1st year after stopping therapy only men who responded in type C fashion were able to impregnate partners, three pregnancies ended in abortion and nine in normal births.

Autoantibodies↗