OBJECTIVES: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. MATERIALS AND METHODS: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from LN2 and letting them at room temperature for 10 min followed by 37degrees C water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. RESULTS: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospermic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. CONCLUSIONS: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.
Baths ; Cryopreservation ; Fertilization ; Humans ; Insemination ; Linear Energy Transfer ; Pregnancy ; Pregnancy Rate ; Seminiferous Tubules ; Sperm Injections, Intracytoplasmic ; Spermatozoa* ; Water

Hexamethonium with propranolol was used to produce hypotension in 3 selected patients, ages 45 to 56, for neck deasection with thyroidectomy, Mile's operration and total laryngectomy with radical neck dissection. These cases were promedicated by lorazepam, anesthesia was induced and intubated following propranolol, morphine, diazepam and alcuroninum, and maintained with halothane, nitrous oxide and oxygen. Respiration was controlled by an anesthesia ventilator. Following elevation of the operation site, small amount of hexamethonium was injected to produce a dry surgical field. Systolic blood pressure was maintained around 80mmHg. Average doses of hexamethonium used were 0.12mg/kghr and propranolol 0.5 to 1 mg respectively. Judicious administration of sufficient doses of propranolol to lower the heart rate before and during hexamethonium induced hypotension reduces both the hexamethonium dose requirement and also the likelihood of developing individual variations in its effect.
Anesthesia* ; Blood Pressure ; Diazepam ; Halothane ; Heart Rate ; Hexamethonium* ; Humans ; Hypotension ; Laryngectomy ; Lorazepam ; Morphine ; Neck ; Neck Dissection ; Nitrous Oxide ; Oxygen ; Propranolol* ; Respiration ; Thyroidectomy ; Ventilators, Mechanical

OBJECTIVES: Klinefelter syndrome is the most common genetic cause of male infertility and presents with 47, XXY mainly or 46, XX/47, XXY mosaicism. It is characterized by hypogonadism and azoospermia due to testicular failure, however, sporadic cases of natural pregnancies have been reported. With the development of testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI), sperm can be retrieved successfully and ART is applied in these patients for pregnancy. It has been suggested that the risk of chromosome aneuploidy for both sex chromosome and autosome is increased in the sperms from 47, XXY germ cells. Considering the risk for chromosomal aneuploidy in the offspring, preimplantation genetic diagnosis (PGD) could be applied as a safe and more effective treatment option in Klinefelter syndrome. The aim of this study is to assess the outcome of PGD cycles by using FISH for sex chromosome and autosome in patients with Klinefelter syndrome. MATERIALS AND METHODS: From Jan. 2001 to Dec. 2003, PGD was attempted in 8 cases of Klinefelter syndrome but TESE was failed to retrieve sperm in the 3 cases, therefore PGD was performed in 8 cycles of 5 cases (four 47, XXY and one 46, XY/47, XXY mosaicism). In one case, ejaculated sperm was used and in 4 cases, TESE sperm was used for ICSI. After fertilization, blastomere biopsy was performed in 6~10 cell stage embryo and the chromosome aneuploidy was diagnosed by using FISH with CEP probes for chromosome X, Y and 17 or 18. RESULTS: A total of 127 oocytes were retrieved and ICSI was performed in 113 mature oocytes. The fertilization rate was 65.3+/-6.0% (mean+/-SEM) and 76 embryos were obtained. Blastomere biopsy was performed in 61 developing embryos and FISH analysis was successful in 95.1% of the biopsied blastomeres (58/61). The rate of balanced embryos for chromosome X, Y and 17 or 18 was 39.7+/-6.9%. The rate of aneuploidy for sex chromosome (X and Y) was 45.9+/-5.3% and 43.2+/-5.8% for chromosome 17 or 18, respectively. Embryo transfer was performed in all 8 cycles and mean number of transferred embryos was 2.5+/-0.5. In 2 cases, clinical pregnancies were obtained and normal 46, XX and 46, XY karyotypes were confirmed by amniocentesis, respectively. Healthy male and female babies were delivered uneventfully at term. CONCLUSION: The patients with Klinefelter syndrome can benefit from ART with TESE and ICSI. Considering the risk of aneuploidy for both sex chromosome and autosome in the sperms and embryos of Klinefelter syndrome, PGD could be offered as safe and more effective treatment option.
Amniocentesis ; Aneuploidy ; Azoospermia ; Biopsy ; Blastomeres ; Chromosomes, Human, Pair 17 ; Embryo Transfer ; Embryonic Structures ; Female ; Fertilization ; Germ Cells ; Humans ; Hypogonadism ; Infertility, Male ; Karyotype ; Klinefelter Syndrome* ; Male ; Mosaicism ; Oocytes ; Pregnancy ; Preimplantation Diagnosis* ; Prostaglandins D ; Sex Chromosomes ; Sperm Injections, Intracytoplasmic ; Spermatozoa

No abstract available.
Humans* ; Pregnancy* ; Sperm Injections, Intracytoplasmic* ; Spermatozoa*

The combination of testicular sperm extraction (TESE) with ICSI can achieve normal fertilization and pregnancy rate and is affective method in obstructive and non-obstructive azoospermic patients. But, when pregnancy was not occurred, repeated testicular biopsies are not evitable. Therefore, in this study, we observed the survival rate of testicular spermatozoa and spermatozoa extracted from the seminiferous tubules after cryopreserved-thawed used for next IVF cycle with ICSI. In a total of 23 cases, obstructive azoospermia was 17 cases and non-obstructive azoospermia was 6 cases. In obstructive azoospermia, after thawing, motile spermatozoa was observed in 13 cases (76.5%). The fertilization rate with 2PN was 67.6% and 5 pregnancies (29.4%) were achieved. In non-obstructive azoospermia, motile spermatozoa was observed in 2 case (33.3%) after thawing. The fertilization rates with 2PN was 53.7% and 3 pregnancies (50.0%) were achieved. A comparison of the results of motile spermatozoa after thawed testicular spermatozoa find spermatozoa extracted from the thawed seminiferous tubule section were 3 cases (60.0%) and 12 cases (86.6%), respectively, The fertilization and pregnancy rates of thawed testicular spermatozoa and spermatozoa extracted from the thawed seminiferous tubule section were 69.4% and 20.0%, 62.5% and 38.8%, respectively. Conclusively, thawed testicular spermatozoa and spermatozoa extracted from the thawed seminiferous tubule section can achieve normal fertilization and pregnancy and cryopreservation of testicular spermatozoa and seminiferous tubule may avoid repetition of testicular biopsies in azoospermic patients in whom the only source of spermatozoa is the testis.
Azoospermia ; Biopsy ; Cryopreservation ; Fertilization ; Humans ; Pregnancy ; Pregnancy Rate ; Seminiferous Tubules* ; Sperm Injections, Intracytoplasmic* ; Spermatozoa* ; Survival Rate ; Testis

This study was carried to determine the possibility of finding motile spermatozoa and fertilization, pregnancy rate after testicular sperm extraction(TESE) with ICSI in obstructive and non-obstructive azoospermic patients. In 154 cases(132 patients), obstructive azoospermia was 77 cases and non-obstructive azoospermia was 77 cases. In obstructive azoospermia, patients generally showed normal spermatogenesis and included vas agenesis(n=8), multiple vas obstruction(n=7), epididymal obstruction (n=54). Total of 982 retrieved oocytes were obtained and 84.4% were injected. The fertilization rates with 2 PN and cleavage rate were 72.5% and 62.3%, .respectively. 30 pregnancies(38.9%) were achieved and the ongoing pregnancies were 22 cases (28.6%). In non-obstructive azoospermia, patients showed hypospermatogenesis(n=49), maturation arrest(n=4), Sertoli cell only syndrome (n=24). The various stages of spermatogenic cell could be retrieved by TESE and could be reached normal fertilization and embryo development with ICSI. Total of 1072 retrieved oocytes obtained and 80.2% were injected. The fertilization rates with 2 PN and cleavage rate were 52.8% and 68.9%, respectively. 22 pregnancies(30.1%) were achieved and the ongoing pregnancies were 19 cases(26.0%). Conclusively, the combination of TESE with ICSI using testicular spermatozoa can achieve normal fertilization and pregnancy rate and effective method in obstructive and non-obstructive azoospermic patients.
Azoospermia ; Embryonic Development ; Female ; Fertilization* ; Humans ; Oocytes ; Pregnancy Rate* ; Pregnancy* ; Sertoli Cell-Only Syndrome ; Sperm Injections, Intracytoplasmic ; Spermatogenesis ; Spermatozoa*

Irreparable obstructive azoospermic patients can be treated successfully with microsurgical epididymal sperm aspiration(MESA) o. testicular sperm extraction (TESE) by intracytoplasmic sperm injection(ICSI). Obstructive azoospermic patients generally have normal spermatogenesis. The aim of this study was to see if any spermatozoa could be retrieved from non-obstructive azoospermia and to assess the efficacy of ICSI with TESE in germinal failure. 42 non-obstructive azoospermic patients revealed no spermatozoa at all in their ejaculates, even after centrifuge. The histology of 42 patients revealed 15 Sertoli cell only Syndrome, 4 maturation arrest and 23 severe hypospermatogenesis. All patients underwent extensive multiple testicular biopsy for sperm retrieval. These patients were scheduled for ICSI using testicular spermatozoa. In 25 out of 42 non-obstructive azoospermic patients, spermatozoa were recovered from multiple testicular biopsy specimen and 11 ongoing pregnancies were achieved. There are usually some tiny foci of spermatogenesis which allow TESE with ICSI in non-obstructive azoospermia. Also these patients may have sufficient sperm in the testes for ICSI, despite extremely high FSH level and small testes.
Azoospermia* ; Biopsy ; Humans ; Male ; Oligospermia ; Pregnancy ; Sertoli Cell-Only Syndrome ; Sperm Injections, Intracytoplasmic ; Sperm Retrieval ; Spermatogenesis ; Spermatozoa ; Testis

Complex chromosome rearrangements (CCRs) are structural chromosomal rearrangements involving at least three chromosomes and more than two breakpoints. CCR carriers are generally phenotypically normal but related to higher risk of recurrent miscarriage and having abnormal offspring with congenital anomalies. However, most of CCR carriers are not aware of their condition until genetic analysis of either abortus or affected baby or parental karyotyping is performed. Herein, we present the case that CCR carrier patients can be identified by preimplantation genetic testing of preimplantation embryos. An infertile male patient with severe oligoasthenoteratozoospermia was diagnosed balanced reciprocal translocation, 46,XY,t(3;11) (p26;p14) at first. After attempting the first preimplantation genetic testing for structural rearrangement (PGT-SR) cycle, we found the recurrent segmental gain or loss on 21q21.3-q22.3 of five out of nine embryos. As a result of karyotype re-analysis, the patient’s karyotype showed a balanced CCR involving chromosomes 3, 11, and 21 with three breakpoints 3p26, 11p14, and 21q21. The patient underwent two PGT-SR cycles, and a pregnancy was established after the transfer of an euploid embryo in the second cycle. Amniocentesis confirmed that the baby carried normal karyotype without mosaicism. At 37 weeks gestation, a healthy girl weighting 3,050 g was born.

OBJECTIVE: To compare the pregnancy outcomes after in vitro fertilization with intracytoplasmic sperm injection (IVF-ICSI) between obstrucvtive and non-obstrucvtive azoospermia. METHODS: From January 1994 to December 2002, 524 patients with obstructive azoospermia (886 cycles) and 163 patients with non-obstructive azoospermia (277 cycles) were included in this study. Microsurgical epididymal sperm aspiration (MESA) or testicular sperm extraction (TESE) in obstructive azoospermia and TESE in non-obstructive azoospermia were perfomed to retrieve sperm, which was used for ICSI and then fertilized embryos were transferred. The results of ICSI-fertlization rate (FR), clinical pregnancy rate (CPR), clinical abortion rate (CAR) and delivery rate (DR)- were statistically analysed in obstructive versus non-obstructive azoospermia. RESULTS: There were no differences in the number of retrieved oocytes, injected oocytes for ICSI and oocyte maturation rate. FR was significantly higher in obstructive than non-obstructive azoospermia (71.7% vs. 61.1%, p<0.001). There was no difference in CPR per embryo transfer cycle. After pregnancy was established, however, CAR was significantly higher in non-obstructive than obstructive azoospermia (25.6% vs. 12.5%, p=0.004). DR per clinical pregnancy cycle was significantly higher in obstructive than non-obstructive azoospermia (78.0% vs. 64.4%, p=0.012). In the karyotype ananlysis of abortus, abnormal karyotypes were found in 75.0% (6/8) of obstructive and 55.6% (5/9) of non-obstructive azoospermia. CONCLUSION: Our data show significantly higher FR in obstructive than non-obstructive azoospermia. Though there was no differrence in CPR, CAR was significantly higher in non-obstructive than obstructive azoospermia. The abortion may be related to the abnormal karyotype of embryo, but further investigations are necessary to elucidate the cause of clinical abortion in azoospermia.
Abnormal Karyotype ; Abortion, Induced ; Azoospermia* ; Cardiopulmonary Resuscitation ; Embryo Transfer ; Embryonic Structures ; Female ; Fertilization in Vitro* ; Humans ; Karyotype ; Oocytes ; Pregnancy ; Pregnancy Outcome* ; Pregnancy Rate ; Pregnancy* ; Sperm Injections, Intracytoplasmic* ; Sperm Retrieval ; Spermatozoa

OBJECTIVE: To determine whether the presence of Y-chromosome microdeletion affects the outcome of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) program. METHODS: Fourteen couples with microdeletion in azoospermic factor (AZF)c region who attempted IVF/ICSI or cryopreserved and thawed embryo transfer cycles were enrolled. All of the men showed severe oligoasthenoteratoazoospermia (OATS) or azoospermia. As a control, 12 couples with OATS or azoospermia and having normal Y-chromosome were included. Both groups were divided into two subgroups by sperm source used in ICSI such as those who underwent testicular sperm extraction (TESE) and those used ejaculate sperm. We retrospectively analyzed our database in respect to the IVF outcomes. The outcome measures were mean number of good quality embryos, fertilization rates, implantation rates, beta-hCG positive rates, early pregnancy loss and live birth rates. RESULTS: Mean number of good quality embryos, implantation rates, beta-hCG positive rates, early pregnancy loss rates and live birth rates were not significantly different between Y-chromosome microdeletion and control groups. But, fertilization rates in the Y-chromosome microdeletion group (61.1%) was significantly lower than that of control group (79.8%, p=0.003). Also, the subgroup underwent TESE and having AZFc microdeletion showed significantly lower fertilization rates (52.9%) than the subgroup underwent TESE and having normal Y-chromosome (79.5%, p=0.008). Otherwise, in the subgroups used ejaculate sperm, fertilization rates were showed tendency toward lower in couples having Y-chromosome microdeletion than couples with normal Y-chromosome. (65.5% versus 79.9%, p=0.082). But, there was no significance statistically. CONCLUSIONS: In IVF/ICSI cycles using TESE sperm, presence of Y-chromosome microdeletion may adversely affect to fertilization ability of injected sperm. But, in cases of ejaculate sperm available for ICSI, IVF outcome was not affected by presence of Y-chromosome AZFc microdeletion. However, more larger scaled prospective study was needed to support our results.
Avena ; Azoospermia ; Embryo Transfer ; Embryonic Structures ; Family Characteristics ; Fertilization ; Fertilization in Vitro* ; Humans ; Live Birth ; Male ; Outcome Assessment (Health Care) ; Pregnancy ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; Y Chromosome*