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Biomedical subjects

F Francavilla

Publications and source records attributed to F Francavilla.

At least 37 records · Page 2Linked to original sources

Interference of antisperm antibodies with the induction of the acrosome reaction by zona pellucida (ZP) and its relationship with the inhibition of ZP binding.

OBJECTIVE: To determine whether antisperm antibodies can interfere with the induction of the acrosome reaction (AR) by the zona pellucida (ZP) and whether this interference also can occur in the absence of an inhibitory effect on ZP binding. DESIGN: Prospective in vitro study. SETTING: A tertiary care center, the Andrologic Clinic, University of L'Aquila. PATIENT(S): Sera from 12 infertile patients with high titers of circulating antibodies directed against the sperm head were studied. INTERVENTION: None MAIN OUTCOME MEASURE(S): The effect of antisperm antibodies on ZP binding was evaluated by matching antibody-exposed and nonexposed donor sperm suspensions labeled with fluorescein or rhodamine, respectively, and incubated with the same salt-stored human ZPs. The effect of antibodies on ZP-induced AR was determined by challenging antibody-exposed and nonexposed donor sperm suspensions with human ZPs disaggregated with acidic NaH2PO4. Acrosomal status was evaluated using fluorescein-labeled Pisum sativum agglutinin and supravital stain Hoechst 33258. In some selected cases, the effect of antisperm antibodies on the acrosomal status of sperm bound to intact ZP also was evaluated using transmission electron microscopy. RESULT(S): Five of 12 sera exhibited an inhibitory effect on ZP binding. An inhibition of AR induction by disaggregated ZPs (ranging from 64% to 98%) was produced by all 5 sera with an inhibitory effect on ZP binding and by 2 of 7 sera without an inhibitory effect on ZP binding. The different effects of antisperm antibodies on AR induction by disaggregated ZP were confirmed by comparing with ultrastructural evaluations on the acrosomal status of sperm bound to intact ZP. CONCLUSION(S): Antisperm antibodies can interfere with the induction of AR by ZP. This inhibition can occur even in the absence of an inhibitory effect on ZP binding. Neither effect may occur.

Acrosome↗

Occurrence of the interference of sperm-associated antibodies on sperm fertilizing ability as evaluated by the sperm-zona pellucida binding test and by the TEST-yolk buffer enhanced sperm penetration assay.

PROBLEM: This study was performed to evaluate the occurrence as well as the level of the interference of sperm-associated antibodies on fertilization process. METHOD: Motile sperm suspensions from 28 infertile patients with high degree of autoimmunization against the sperm head were tested with the zona pellucida (ZP) binding test and with the sperm penetration assay (SPA) enhanced with TEST-yolk buffer. Both tests were also performed using donor sperm exposed and non-exposed to the patients' circulating sperm antibodies. RESULTS: A low ZP-binding was exhibited by sperm from 50% of patients with normal semen profile. All normozoospermic patients with low ZP-binding showed circulating sperm-antibodies with inhibitory effect on ZP-binding, while no patient with normal ZP-binding showed circulating sperm-antibodies with inhibitory effect. No normozoospermic patient exhibited a negative SPA result, and only in 16% of cases the penetration index was slightly less than 2 (the lowest value exhibited by fertile controls). Circulating antisperm-antibodies did not significantly affect the hamster egg penetration. CONCLUSION: Even in the presence of high degree of autoimmunization against the sperm head, sperm fusion with oolemma is not impaired after sperm preincubation with TEST-yolk buffer, while an impairment of the ZP-binding is demonstrable in half cases, when non-immunologic factors are excluded. A substantial role in this interference is likely exerted by IgG antibodies transuded from the blood into the genital tract.

Autoantibodies↗

Relationship between acrosome reactions and hamster egg penetration after ionophore challenge in absence of teratozoospermia.

OBJECTIVE: To determine the relationship between acrosome reactions and hamster egg penetration after ionophore challenge in nonteratozoospermic semen samples. SETTING: A tertiary care center, the Andrologic Clinic, Department of Internal Medicine, University of L'Aquila. PATIENTS: Twenty-five nonteratozoospermic patients with infertile marriages and nine fertile men. INTERVENTIONS: The hamster egg penetration assay and the acrosome reaction assessment were performed on capacitated sperm suspensions in basal conditions and after ionophore challenge with ionomycin. MAIN OUTCOME MEASURE: The relationship between the acrosome reactions and the hamster egg penetration was analyzed. RESULTS: Although in basal conditions the spontaneous acrosome reaction rate was not correlated with the hamster egg penetration, after ionophore challenge both the induced acrosome reaction rate and the acrosome reaction increase (induced minus spontaneous acrosome reaction rate) correlated with the hamster egg penetration. The highest correlation was observed between the induced acrosome reaction rate and the penetration index. Considering a penetration index = 2 as the threshold of a good response of the hamster egg penetration assay to the ionophore challenge, significantly lower acrosome reaction rates and increases were associated with poor hamster egg penetration. However, different penetration indexes occurred at the same level of induced acrosome reaction rate. CONCLUSIONS: The capability of sperm to react after ionophore challenge reflects to some extent the capability of the acrosome-reacted sperm to undergo the changes of the plasma membrane that are necessary for sperm-oocyte fusion. The simple evaluation of the acrosome reactions may represent a useful complement rather than a substitute for the hamster egg penetration assay in monitoring the responses of human sperm to the ionophore challenge. An impaired inducibility of the acrosome reactions may account for poor hamster egg penetrations exhibited by nonteratozoospermic semen samples.

Acrosome↗

Impaired hamster egg penetration by human sperm from ejaculates with low acrosin activity but otherwise normal.

OBJECTIVE: To determine if a low acrosin activity in otherwise normal ejaculate from infertile patients is associated with an impairment of sperm functions involved in the hamster egg penetration. SETTING: A tertiary care center, the Andrology Clinic, Department of Internal Medicine, University of L'Aquila. PATIENTS: Nine infertile patients with low acrosin activity in otherwise normal ejaculate (including normal immunoreactivity for acrosin) were studied; nine fertile men served as a control group. The two groups were homogeneous for seminal parameters. INTERVENTIONS: The hamster egg penetration assay and the acrosome reaction rate assessment were performed on capacitated sperm suspensions in basal conditions and after ionophore challenge. MAIN OUTCOME MEASURES: The penetration rate, the penetration index, and the acrosome reaction rate were measured and compared between the groups. RESULTS: No penetration was achieved in seven patients and a low penetration was achieved in two cases. The difference with the control group was significant. The ionophore challenge was associated to penetration of hamster eggs in seven of nine patients, but the penetration index was significantly lower than the controls. Acrosin activity was correlated to hamster egg penetration. Both spontaneous and induced acrosome reaction rate were similar in the two groups. CONCLUSIONS: A low sperm acrosin activity in otherwise normal ejaculate is associated with an impaired hamster egg penetration. This impairment does not seem to be due to altered dynamics of acrosome reaction.

Acrosin↗

Sperm-zona pellucida binding of human sperm is correlated with the immunocytochemical presence of proacrosin and acrosin in the sperm heads but not with the proteolytic activity of acrosin.

OBJECTIVE: To determine whether the sperm-zona pellucida (ZP) binding is related to the presence of immunoreactivity for proacrosin and acrosin and/or to the proteolytic activity of acrosin. SETTING: The Andrology Clinic, University of L'Aquila, Italy. PATIENTS: Thirty-five infertile couples and 15 men of proven fertility. INTERVENTIONS: Salt-stored unfertilized oocytes after IVF were inseminated with a mixture of equal numbers of test and fertile donor sperm after swim-up selection, respectively labeled with fluorescein or rhodamine. Total acrosin activity, the percent of sperm immunostained for proacrosin and acrosin, and the percent of sperm with oval heads were evaluated in fresh semen and after swim-up selection. MAIN OUTCOME MEASURES: Three zonae were used for each patient and sperm-ZP binding ratio (ZP ratio) was calculated as the total number of test sperm bound to ZP divided by the total number of control sperm bound to ZP. The ZP ratio was compared with the percent of live sperm immunostained for proacrosin and acrosin, with total acrosin activity and with the percent of sperm with oval heads after swim-up selection. RESULTS: The sperm-ZP binding ratio was correlated positively with the percent of sperm immunostained for proacrosin and acrosin and with the percent of sperm with oval heads, but not with acrosin activity. Ejaculates with immunostaining for proacrosin and acrosin > or = 55% of sperm (the mean value of infertile couples) and acrosin activity < 24 muIU/10(6) sperm (the mean value of infertile couples) (n = 8) had a ZP ratio > or = 0.5, whereas ejaculates with low immunostaining for proacrosin and low acrosin activity (n = 12) had a ZP ratio < 0.5. The difference between the ZP ratio of the two groups was highly significant. CONCLUSIONS: The ZP binding is related to the immunocytochemical presence of proacrosin and acrosin but not to the proteolytic activity of acrosin in human sperm, suggesting that egg recognition and proteolytic activity are independent functions of proacrosin and acrosin.

Acrosin↗

Effect of ionophore challenge on hamster egg penetration and acrosome reaction of antibody-coated human sperm.

PROBLEM: Following the demonstration that antisperm antibodies do not affect the spontaneous acrosome reactions (AR) of human sperm used for the hamster egg penetration assay (HEPA), we evaluated the effect of the ionophore challenge on HEPA and AR of antibody-coated sperm. METHOD: Motile sperm suspensions from donors were exposed to serum and seminal plasma samples with sperm-head-directed antibodies, washed, capacitated, and challenged with ionomycin. RESULTS: The proportion of acrosome-reacted sperm, assessed by fluoresceinated Pisum sativum agglutinin and supravital stain Hoechst 33258, was significantly higher in the antibody-exposed sperm than in the controls (47.0 +/- 13.2% vs. 39.3 +/- 12.1, respectively; P < 0.05). However, the hamster egg penetration rate and index were not significantly different between antibody-exposed sperm and controls (98.0 +/- 3.7% vs. 85.9 +/- 16.2% and 6.64 +/- 4.1 vs. 6.81 +/- 4.8, respectively). Only one of eight test samples with antisperm antibodies, where the highest increase of AR rate compared with controls was observed (55.0% vs. 35.6%) produced a substantial increase of penetrations per oocyte (7.4 vs. 4.1). CONCLUSION: The ionophore challenge enabled us to recognize a subtle effect of anti-sperm antibodies on the dynamics of acrosomal loss, the clinical relevance of which needs to be elucidated.

Acrosome↗

The effect of a single course of alpha-2B-interferon in patients with HIV-related and chronic idiopathic immune thrombocytopenia.

13 patients with HIV-related immune thrombocytopenia (HIV-ITP) and 10 patients with chronic idiopathic thrombocytopenic purpura (C-ITP) were treated with a single course of alpha-2b-Interferon (IFN 3 x 10(6) IU subcutaneously for 12 d). The patients had platelet counts lower than 40 x 10(9)/L and thrombocytopenia persisting for over 1 year (range 1-22 years); 7 patients were refractory to previous conventional therapy, 5 were responsive, and 11 had not been previously treated. The response to IFN was complete in 8 patients (platelets > 100 x 10(9)/L), partial in 7 (platelets 50-100 x 10(9)/L); 8 patients showed no response. The treatment with IFN was stopped after 4 d in one patient due to a fall in platelet count. The maximal platelet count (median peak 116 +/- 55 SD x 10(9)/L platelets) was obtained after 13.7 +/- 2.98 d and the improvement in platelet count was maintained for 22.8 +/- 8.6 d. No difference in platelets response was observed between HIV-ITP and C-ITP. The response to IFN seems to be related to the one obtained with previous treatments. Indeed 80% of the patients who were responsive to previous steroids, high dose immunoglobulins or azidothymidine (HIV-ITP) showed a complete or partial response while only 43% of the refractory patients showed a partial response; the positive response rate in previously untreated patients was 73%.

Adult↗

Failure of intrauterine insemination in male immunological infertility in cases in which all spermatozoa are antibody-coated.

OBJECTIVE: To determine if the overcoming of the cervical mucus barrier removes the interference of sperm-bound antibodies with fertility. DESIGN: Prospective case series. SETTINGS: University-based intrauterine insemination (IUI) homologous program. PATIENTS: Nineteen patients with all spermatozoa in the ejaculate coated by antisperm antibodies. As control group, 86 consecutive patients without antisperm antibodies, treated for oligoasthenozoospermia or mucus hostility. INTERVENTIONS: Intrauterine inseminations (at least 3 attempts per couple). MAIN OUTCOME MEASURES: The outcome of IUIs, demographic, and seminal parameters were compared between the two groups. RESULTS: No pregnancy occurred in the couples with male immunological infertility, treated by 110 IUIs. Twenty-three pregnancies occurred in 22 (25.6%) of the control group couples who were treated by 411 IUIs. In the group of patients without antisperm antibodies, we demonstrated that the pregnancy rate (PR)/couple in oligoasthenozoospermia without teratozoospermia was similar to that achieved in normozoospermia (35% versus 38.9%), whereas it was significantly affected by teratozoospermia (3.6%). Only three patients with antisperm antibodies had teratozoospermia. Comparing the PR per couple and per cycle between the two groups of patients (with and without antisperm antibodies), excluding the patients with teratozoospermia, significant differences resulted (P less than 0.005 and P less than 0.005, respectively). The motile sperm count was not significantly different between the two groups, which also resulted to be homogeneous for demographic data. Moreover, the motile sperm count was not different between the patients with and without antisperm antibodies, who had successful IUI. CONCLUSIONS: The analysis of this trial suggests that the failure of IUI in the treatment of male immunological infertility is imputable to antisperm antibodies when they involve all spermatozoa, regardless of semen quality.

Autoantibodies↗

Effect of sperm-antibodies on acrosome reaction of human sperm used for the hamster egg penetration assay.

The effect of anti-sperm antibodies (ASA) on the rate of acrosome reactions (AR) during "in vitro" capacitation of human sperm used for the hamster egg penetration assay (HEPA) was assessed. Motile sperm suspensions from donors were exposed to several sera and seminal plasma with sperm head-directed ASA, then they were washed and capacitated "in vitro." After capacitation, the proportion of acrosome-reacted viable sperm was assessed by staining with Fluoresceinated Pisum Sativum Agglutinin and supravital stain Hoechst 33258. ASA of any immunoglobulin class did not significantly affect either the AR rate, or the hamster egg penetration rate. In conclusion, interference of ASA on spontaneous AR rate during "in vitro" capacitation can not be advocated as an explanation of the impairment of the interaction of human sperm with egg or its vestments, which have been reported in several studies.

Acrosome↗

Effect of sperm morphology and motile sperm count on outcome of intrauterine insemination in oligozoospermia and/or asthenozoospermia.

Eighty-six couples with long-standing infertility and poor postcoital test, due to oligozoospermia and/or asthenozoospermia (68 cases) or mucus hostility (18 cases), were treated by 411 intrauterine inseminations (IUI) with motile sperm suspensions from the husband's semen. The pregnancy rate per couple in the group with abnormal semen was lower than in the group with mucus hostility (22% versus 38.9%). Influence of seminal and other parameters on outcome of IUI was assessed by discriminant analysis, and a significant correlation with pregnancy rate was found for motile sperm count and sperm morphology. Teratozoospermia (normal morphology less than 50%) affected the outcome of IUI both when associated with moderate oligozoospermia and/or asthenozoospermia (motile sperm count greater than or equal to 5 X 10(6)/mL) (success rate per couple: 11.1%), and, even more, when associated with severe oligozoospermia and/or asthenozoospermia (motile sperm count less than 5 X 10(6)/mL), where no pregnancy was achieved. In the absence of teratozoospermia, the success rate per couple both in severe and in moderate oligozoospermia and/or asthenozoospermia had similar results (33.3% versus 35.7%). In conclusion, the absence of teratozoospermia appears to be an effective criterion for selecting couples with infertility due to oligozoospermia and/or asthenozoospermia who may benefit from IUI.

Adult↗

Quantitative evaluation of Leydig cells in testicular biopsies of men with varicocele.

A quantitative analysis of Leydig cells was performed in 23 testicular biopsies of men with left varicocele and sperm count ranging from zero to 95,000 sperm/mm3. The oligozoospermic patients had more Leydig cells and higher FSH and LH serum levels than the patient group with more than 10,000 sperm/mm3. The Leydig cell density appeared tightly correlated (p less than 0.01) with the serum level of LH. In oligozoospermic subjects, an altered Leydig cell function could trigger an increased LH secretion; this seems likely to be responsible for the stimulation of interstitial cells resulting in an exaggerated recruitment of mature Leydig cells from their precursors. The comparative analysis of left and right testes failed to show differences in Leydig cell density and spermatogenesis in normozoospermic and oligozoospermic patients. This suggests that the two testes are equally involved by a possible, although unknown, detrimental effect of left side varicocele.

Adult↗

Norfloxacin concentrations in seminal plasma measured by HPLC.

Eighteen healthy volunteers aged 22 to 37 (average 30.44 +/- 4.11 years) seeking fertility evaluation at the Fertility and Sterility Centre of the Medical Clinic of the University of L'Aquila Medical School were studied for norfloxacin concentrations in serum, urine and seminal plasma 14-15 h after taking two 400 mg doses of the drug. The extraction of the biological fluids and the high pressure liquid chromatographic (HPLC) assay were performed following a method described previously (1), with seminal plasma samples processed in the same way as serum samples. The results in urine and serum substantially confirmed the figures already established for norfloxacin concentrations in such fluids. Seminal plasma levels (average 4.78 +/- 2.69 micrograms/ml) were about 5 times higher than the corresponding serum levels (average 0.89 +/- 0.59 microgram/ml). The authors believe that such concentrations, attaining a therapeutic range in the seminal plasma after only two 400 mg doses of norfloxacin, support its use as an effective chemotherapeutic agent in the management of male genital infections that frequently account for subfertility conditions in young patients.

Adult↗

Acute cadmium intoxication: influence of cyproterone acetate on the testis and epididymis of the rat.

The changes resulting from treatment with cadmium were studied following the histological changes, the modification of both vascular permeability to vital dyes and of alkaline phosphatase activity in rat testis and epididymis. The testicular extravasation of acriflavine started 90 min following parenteral injection of cadmium and increased thereafter synchronous with an increase in testicular and epididymal weights due to edema. At 14 and 24 hr a striking decrease of interstitial fluorescence and tubular degeneration were noted in testis and caput epididymis due to thrombosis of the microvascular circulation. The barrier noted at 8 hr following cadmium injection. No changes of alkaline phosphatase activity was detected in testicular and epididymal blood vessels after cadmium injection. Previous treatment with cyproterone acetate accelerated the appearance of such alterations. The interstitial nuclear staining with acriflavine appeared in the testis at 1 hr and was diffuse at 90 and 120 min. cyproterone acetate seemed to accelerate the appearance of tubular degeneration at 8 hr after cadmium injection. The changes of the male rat gonad following cadmium treatment were characterized by an increased vascular permeability and generalized thrombosis. An inbalance of androgen stimulation seems to increase the blood vessels susceptibility to cadmium.

Acriflavine↗

[Gonadotropin response to slow infusion of GN-RH in idiopathic ologospermia. Preliminary findings].

Gn-RH was administered i.v. to infertile patients presenting with idiopathic oligospermia at a constant rate (0,4 microgram) for 4 h. All the patients showed an increased FSH response, and LH levels higher than normal in both the first and the second peak of response. Our data indicate that an increased pituitary sensitivity and reserve of FSH and LH are present in idiopathic oligospermia, suggesting an involvement of both Leydig cells and seminiferous epithelium in this condition.

Adult↗

[Gonadotropin response to acute stimulation with LHFSHRH in idiopathic oligospermia. Preliminary results].

Serum FSH and LH levels in basal conditions and after Gn-RH test were investigated in 15 idiopathic oligozoospermic patients. Basal FSH was significantly higher in oligospermic than in normal subjects and a negative relationship was found between basal FSH and sperm count. Basal LH was not different in oligozoospermic and in normal subjects. Both FSH and LH responses to GN-RH were significantly higher in oligozoospermic patients. In idiopathic oligozoospermia the presence of a testicular defect involving both the seminiferous epithelium and Leydig cells is suggested.

Adult↗

[Development and differentiation of the rat epididymis. I: ultrastructural aspects of the peritubular zone].

We investigated the ultrastructural aspects of the peritubular cells of epididymis and their development from birth to adult age. At birth the peritubular zone consisted of polygonal cells which did not differ from other interstitial cells. Cytoplasmic filaments were visible in the cells of the inner layer at day 6. From day 22 the peritubular cells reached the adult aspect. The peritubular cells in the rat epididymis had aspects similar to those of peritubular smooth muscle cells of rat testis, with a more precocious appearance of cytoplasmic filaments. This finding concurs with the observed precocious contractility of epididymis.

Animals↗

[Development and differentiation of the rat epididymis. II: histochemical aspects of the peritubular zone].

Development of the contractile peritubular structures of rat testis, from birth to full maturation, was investigated by histochemical evaluation of alkaline phosphatase activity (A.P.A.) at 0, 16, 22 e 35 days of age. A.P.A. appeared at 6 days, age of appearance also of the cytoplasmic microfilaments of peritubular cells. The correlation between cytoplasmic microfilaments and A.P.A. was confirmed by the pattern of A.P.A. positivity at 6 degree day, when the inner peritubular cells layer, in which the microfilaments are present, are more positive than the outer layer in which the microfilaments are not yet present. These findings give further support to the belief that the cytoplasmic microfilaments of peritubular cells are contractile structures.

Alkaline Phosphatase↗

Ultrastructural changes in the seminiferous tubule wall and intertubular blood vessels in human cryptorchidism.

Biopsy material was obtained from adult men whose both testes had descended into the scrotal sac either spontaneously by use of pharmacologic agents or through surgery at age 7--16 yr, and from undescended testes obtained during orchipexy in men 8--40 yr of age. Testes appeared normal until the 10th yr, when progressive alterations were observed with the onset of puberty and through the adult age both in retained testes and cases of testes that descended through chemical or surgical methods before or after puberty. A thickening of the inner and outer acellular layers of the seminiferous tubule wall was due to an accumulation of collagen fibers and microfibrillar material. The basement membrane was also thickened and appeared as a homogeneous or a multilaminar structure. The intertubular blood vessels presented a thickening of the tunica media and a laminar accumulation of basement-membrane-like material.

Adolescent↗