Kazuo Saito

Effect of reactive oxygen species in semen on the pregnancy of infertile couples.

Objective:u2003 We intended retrospectively to investigate whether reactive oxygen species (ROS) levels, detected in whole semen, were correlated with the actual pregnancy rate.

Successful pregnancy with sperm retrieved from the bladder after the introduction of a low-electrolyte solution for retrograde ejaculation.

OBJECTIVEnTo determine the safety and efficacy of the introduction of a low-electrolyte solution into the bladder before masturbation for sperm recovery in patients with retrograde ejaculation.nnnDESIGNnTwo case reports.nnnSETTINGnAssisted reproduction practice at a university hospital.nnnPATIENT(S)nTwo men with retrograde ejaculation.nnnINTERVENTION(S)nA 0.32-mol/L glucose solution or Hams modified F-10 medium (GIBCO, Grand Island, NY) with 10% serum was introduced into the urinary bladder before masturbation. The retrieved sperm were used for IUI.nnnMAIN OUTCOME MEASURE(S)nThe motility of retrieved sperm, achievement of pregnancy, and outcome of pregnancy.nnnRESULT(S)nIn patient 1, the motility rate of sperm retrieved from the bladder after the introduction of Hams modified F-10 medium was higher than that after the introduction of a glucose solution. The patients wife did not become pregnant during 10 IUI cycles after treatment with Hams modified F-10 medium. However, she became pregnant during the first IUI cycle after treatment with a glucose solution and was delivered of healthy twin infants. The wife of patient 2 also became pregnant during the first IUI cycle after treatment with a glucose solution and was delivered of a healthy female infant.nnnCONCLUSION(S)nThe introduction of a glucose solution appears to be a safe and simple method for sperm recovery in patients with retrograde ejaculation.

A new method of the electrolyte-free long-term preservation of human sperm at 4°C

Objectives To develop a new method for the long-term preservation of human sperm. Setting Andrology laboratory of our hospital. Patients Thirty-one normal and 19 asthenozoospermie semen samples obtained from patients attending our infertility clinic. The average sperm motility was 70.2% and 36.0% in the normal and asthenozoospermie groups, respectively. Interventions Ejaculated sperm were centrifuged and washed using the electrolyte-free Percoll gradient and then were preserved at 4°C. Main Outcome Measures The motility of the preserved sperm was analyzed using computer-assisted semen analyzer after the addition of Hams modified F-10 (GIBCO, Grand Island, NY). Results In the normal group, motility rate after the addition of Hams F-10 for 1, 2, and 4 weeks of preservation was 65.4%, 40.4%, and 5.5%, respectively. In the asthenozoospermie group, motility rate after 1 and 2 weeks of preservation was 31.3% and 18.1%, respectively. Preservation solutions containing sodium or potassium decreased motility after preservation. Restoration of preserved sperm was not achieved by incubation alone; however, reinitiation was induced by incubation together with Hams F-10. Conclusions Human sperm in the electrolyte-free solution survived for a long period of time at 4°C and reinitiation of sperm motility after preservation required the addition of Hams F-10.

The role of potassium ion and extracellular alkalization in reinitiation of human spermatozoa preserved in electrolyte-free solution at 4°C

OBJECTIVEnTo elucidate reinitiation factors in human spermatozoa preserved in the electrolyte-free solution at 4 degrees C.nnnSETTINGnAndrology laboratory of our hospital.nnnPATIENTSnSemen samples were obtained from patients attending our infertility clinic.nnnINTERVENTIONSnEjaculated sperm were centrifuged and washed using the electrolyte-free Percoll gradient and then were preserved for 1 week at 4 degrees C.nnnMAIN OUTCOME MEASURESnThe motility of preserved sperm was incubated and analyzed after the addition of electrolyte solutions.nnnRESULTSnThe motility rate was 9.6% when the preserved sperm were incubated directly. The motility increased to 35.1% after alkalization (pH 7.8) and further increased to 40.7% when 0.1 mM KCl was added. The motility decreased to 1.4% in a weak acidic solution (pH 6.8), however, the addition of a > or = 10 mM concentration of NaCl or > or = 0.1 mM concentration of KCl increased in motility. The motility rate in 40 mM NaCl and 40 mM KCl (pH 6.8) was 19.8% and 31.5%, respectively. The restoration of motility by NaCl was inhibited by 1 mM amiloride. The motility also rose to 54.1% and 32.3% in 0.04% NH3 and 80 mM NH4Cl solution, respectively.nnnCONCLUSIONSnThe reinitiation of preserved spermatozoa was induced by potassium ion and extracellular alkalization.

Direct and indirect effects of electrical stimulation on the motility of human sperm

Background: Electroejaculation is a technique that is becoming more frequently available for obtaining sperm from men with spinal cord injury or psychogenic anejaculation. However, the effect of electrical stimulation on the movement of human sperm has not been clarified.

Viability and function of human sperm in electrolyte-free cold preservation

OBJECTIVEnTo assess the viability and function of human sperm in electrolyte-free cold preservation.nnnDESIGNnProspective comparative study.nnnSETTINGnAndrology laboratory of our hospital.nnnPATIENT(S)nTen semen samples obtained from patients attending our infertility clinic.nnnINTERVENTION(S)nEjaculated sperm were washed using the electrolyte-free Percoll gradient and were then preserved in 0.33 M glucose solution, 0.16 M NaCl solution, 0.16 M KCl solution at 4 degrees C for 4 weeks. As a control, TEST (TES and Tris) yolk buffer (TYB) was added to the ejaculated semen and preserved at 4 degrees C.nnnMAIN OUTCOME MEASURE(S)nSperm tail morphology, motility, viability (eosin-Y stain), and the concentration of adenosine triphosphate (ATP) were analyzed.nnnRESULT(S)nThe number of sperm with normal tail form and the motility of sperm preserved in glucose solution (electrolyte-free cold preservation) were significantly (P < 0.01) higher for 4 weeks than those of sperm preserved in the other three media. The sperm viability in glucose solution was 75.5%, 65.4%, and 51.3%, after 1, 2, and 4 weeks, respectively. The ATP concentration after 1, 2, and 4 weeks remained 64.2%, 53.0%, and 4.3% of the prestorage value, respectively, in the sperm stored in glucose solution.nnnCONCLUSION(S)nThe morphology, motility, viability, and ATP concentration of sperm in electrolyte-free cold preservation were substantially better than those in NaCl solution, KCl solution, or TYB for 2 weeks.

Successful pregnancy with intrauterine insemination using vasal sperm retrieved by electric stimulation

OBJECTIVEnTo determine the feasibility of electric stimulation applied to the epididymis or vas deferens to retrieve vasal sperm.nnnDESIGNnTwo case reports.nnnSETTINGnAssisted reproduction practice in a hospital.nnnPATIENT(S)nTwo patients with retrograde ejaculation and severe asthenozoospermia.nnnINTERVENTION(S)nPulse electric simulation was applied to the epididymis or vas deferens to retrieve sperm via a tube cannulated into the vas deferens. After diluting with culture medium, retrieved sperm were used for IUI.nnnMAIN OUTCOME MEASURE(S)nThe volume of retrieved vasal fluid and motility parameters of the sperm, the achievement of pregnancy, and outcome of pregnancy.nnnRESULT(S)nIn patient 1, the volumes of vasal fluids and numbers of sperm were 10-40 microL and 31.4-75.9 x 10(6), respectively, during two cycles. Sperm motility was 88.4%--93.2%. His wife became pregnant and was delivered of a healthy infant by IUI during the second cycle. In patient 2, 0.6 x 10(6) sperm with a motility of 70% were retrieved and used for IUI. However, his wife did not become pregnant.nnnCONCLUSION(S)nElectric vasal sperm retrieval is a feasible method for collecting sperm from the vas deferens in selected patients with male factor infertility.

Effects of extracellular ions on the reactivation of human spermatozoa preserved in electrolyte-free solution

It has previously been reported that human spermatozoa preserved in an electrolyte‐free solution can survive for several weeks at 4u2003°C. However, the motility of spermatozoa after preservation cannot be restored when incubated at 37u2003°C, unless reactivated by extracellular alkalisation. Under weak acidic conditions, the reactivation is induced by ≥10u2003mmolu2003l–1 Na+ and inhibited by a Na+‐H+ antiporter inhibitor. The addition of ≥0.1u2003mmolu2003l–1 K+ also induces the reactivation. In the present study, the reactivation was induced by ≥0.1u2003mmolu2003l–1 Rb+ or ≥1u2003mmolu2003l–1 Cs+ at an acidic pH. The maximum motility rate with K+, Rb+ or Cs+ was obtained at 10–20u2003mmolu2003l–1 and inhibited by 10−5–10−2 molu2003l−1 ouabain in a dose‐dependent manner, while ouabain had no effect on the Na+‐induced reactivation. The addition of K+ further increased sperm motility reactivated by Na+, which was also inhibited by ouabain. The addition of Ca2+ did not induce the reactivation or increase sperm motility reactivated by Na+ or K+. It was concluded that activation of the ouabain‐sensitive Na+‐K+‐ATPase and Na+‐H+ exchange mechanism has an important role in sperm motility.

A Case of Gross Hematuria Arising during Embolization for Renal Arteriovenous Malformation

A cirsoid type of arteriovenous malformation was treated successfully by superselective embolization using a platinum micro coil and absorbable gelatin sponge. During the procedure, as soon as the arteriovenous shunt had disappeared, an arteriopelvic shunt appeared. The pyelogram showed that the high pressure arterial blood flow drained directly into the pelvic cavity. Massive gross hematuria occurred immediately accompanied by a sudden blood pressure drop to 60 mm Hg, necessitating blood transfusion. Fortunately this complication was managed successfully. To our knowledge, there are only a few reported cases that make reference to gross hematuria occurring on embolization. We discuss the possible causes of this complication and the mechanism of occurrence of gross hematuria in renal arteriovenous malformation.