• Clinical and Experimental Reproductive Medicine
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• v.36 no.4
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• pp.301-310
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• 2009

Objective: To compare the embryonic development and pregnancy results using sperms retrieved from fresh and frozen-thawed testicular tissue in patients with obstructive (OA) and non-obstructive azoospermia (NOA). Methods: A total two hundred twenty-two cycles of TESE-ICSI were performed in OA and NOA. Sperms were retrieved from fresh and frozen-thawed testicular tissue. ICSI was performed patient's own sperm. Fertilization was assessed 16~18 hrs after ICSI. Embryo development and pregnancy rates were analysed. Results: The fertilization rates were significantly different between OA and NOA patients (75.2% vs. 56.7%, p<0.05), however, embryo development did not differ between the groups (96.9% vs. 98.0%). Likewise, OA and NOA groups had no differences in their clinical pregnancy and delivery rates, 33.9% vs. 36.0% and 28.1% vs. 28.0%, respectively. With regard to sperm retrieved from fresh testicular tissue, fertilization rates were significantly different between the OA and NOA groups (76.4% vs. 52.9%, p<0.05); however, embryo development, clinical pregnancy and delivery rates were not different. For sperm retrieved from thawed testicular tissue, the fertilization rates were significantly different between the two groups (74.7% OA group vs. 65.6% NOA group, p<0.05); however, embryo development, clinical pregnancy and delivery rates were not different. Conclusions: Embryo development and clinical pregnancy did not differ in patients with obstructive and non-obstructive azoospermia, whether sperm retrieved from fresh and thawed testicular tissue were used, although the fertilization rates were different. Therefore, ICSI with sperm retrieved from fresh and thawed testicular tissue could achieve relevant clinical pregnancy results in patients with azoospermia.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.2
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• pp.155-160
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• 2001

Objective: ICSI with testicular sperm could achieve optimal fertilization and pregnancy. This study was performed to observe the influence on fertilization and pregnancy of motility of fresh testicular sperm and sperm extracted from frozen-thawed seminiferous tubules in obstructive azoospermia. Materials and Methods: We analysed clinical outcome of ICSI using fresh testicular sperm and sperm extracted from thawed seminiferous tubules. The presence of motility were compared to determine the factor for optimal fertilization and pregnancy rates. Results: In 316 cases of TESE-ICSI in obstructive azoospermia, ICSI with fresh testicular sperm (fresh sperm group) were 163 cases and ICSI with sperm testicular sperm extracted from frozen-thawed seminiferous tubule (thawed sperm group) were 153 cases. The fertilization rates were 71.3% and pregnancy rates were 32.5% in fresh sperm group, in thawed sperm group, 65.1% and 33.3% respectively. The fertilization and pregnancy rates of motile and non-motile testicular sperm were 72.9% and 33.6%, 50.0% and 18.2%, respectively (p<0.05). The fertilization and pregnancy rates of motile and non-motile sperm extracted from the thawed seminiferous tubule were 67.8% and 34.7%, 55.1% and 28.1%, respectively (p<0.05). The comparative of the results of ICSI using motile fresh testicular sperm and motile sperm extracted from thawed seminiferous tubule, fertilization and pregnancy rates were not significantly different (72.9% and 33.6%, 67.8% and 34.7%, respectively). Conclusion: These results suggest that successful pregnancy in TESE-ICSI treatment is influenced by the motility of fresh testicular sperm and sperm extracted from thawed seminiferous tubule in obstructive azoospennic patients.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.3
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• pp.207-215
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• 2003

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.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.2
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• pp.45-50
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• 2015

Objective: Artificial oocyte activation (AOA) is an effective method to avoid total fertilization failure in human in vitro fertilization-embryo transfer (IVF-ET) cycles. AOA performed using a calcium ionophore can induce calcium oscillation in oocytes and initiate the fertilization process. We evaluated the usefulness of AOA with a calcium ionophore in cases of total fertilization failure in previous cycles and in cases of severe male factor infertility patients with non-motile spermatozoa after pentoxifylline (PF) treatment. Methods: The present study describes 29 intracytoplasmic sperm injection (ICSI)-AOA cycles involving male factor infertility at Cheil General Hospital from January 2006 to June 2013. Patients were divided into two groups (control, n=480; AOA, n=29) depending on whether or not AOA using a calcium ionophore (A23187) was performed after testicular sperm extraction-ICSI (TESE-ICSI). The AOA group was further split into subgroups according to sperm motility after PF treatment: i.e., motile sperm-injected (n=12) and non-motile sperm-injected (n=17) groups (total n=29 cycles). Results: The good embryo rate (52.3% vs. 66.9%), pregnancy rate (20.7% vs. 52.1%), and delivery rate (10.3% vs. 40.8%) were lower in the PF/AOA group than in the control group. When evaluating the effects of restoration of sperm motility after PF treatment on clinical outcomes there was no difference in fertilization rate (66.6% vs. 64.7% in non-motile and motile sperm, respectively), pregnancy rate (17.6% vs. 33.3%), or delivery rate (5.9% vs. 16.7%) between the two groups. Conclusion: We suggest that oocyte activation is a useful method to ensure fertilization in TESE-ICSI cycles regardless of restoration of sperm motility after PF treatment. AOA may be useful in selected patients who have a low fertilization rate or total fertilization failure.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.2
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• pp.171-177
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• 1998

The combination of testicular sperm extraction (TESE) with ICSI can achieve normal fertilization and pregnancy rate and is effective 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 spemratozoa 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 spermatozua 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 and spermatozoa extracted from the thawed seminiferous tubule section were 3 cases (60.0%) and 12 cases (66.6%), 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.

• Journal of Microbiology and Biotechnology
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• v.11 no.1
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• pp.29-34
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• 2001

The mutant enzymes of N-carbamyl-D-amino aicd amidohydrolase (N-carbamylase) from Agrobacterium radiobacter NRRL B11291, showing a negligible activity, were selected from the library generated by random mutagenesis. From the sequence analysis, these mutants were found to contain the amino acids substitutions at Cys172, Glu47, and Glu146. Previously, Cys172 was reported to be necessary for the enzyme catalysis. The chemical modification of the N-carbamylase by carboxyl group specific chemical reagent, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide(EDC), resulted in a loss of activity. The replacement of glutamic acids with glutamines by site-directed mutagenesis led to aggregation of the enzymes. Mutant enzymes fused with maltose binding protein (MBP) were expressed in soluble form, but were inactive. These results indicate that two glutamic acid residues play an important role in structure and function of the N-carbamylase. Multiple sequence alignment of the related enzymes revealed that Glu47 and Glu146 are rigidly conserved, which suggests that tese residues are crucial for the structure and function of the functionally related C-N hydrolases.

• Journal of Embryo Transfer
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• v.19 no.1
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• pp.11-18
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• 2004

The aim of this study was to evaluate the improvement of testicular sperm motility following different culture conditions such as human follicular fluid (hFF) and temperature. Testicular tissues obtained from azoospermia (n=21) were minced into small pieces by blade and recovered sperm suspension were cultured in Ham's F10 with or without 40% hFF at different temperatures (Group I: 37$^{\circ}C$/with hFF, Group II: 32$^{\circ}C$/withGroup III: 37$^{\circ}C$/without, Group IV:32$^{\circ}C$ /without The motility and viability of sperm were monitored during culture for 48 hours. Initial motility of testicular sperm was 10.9$\pm$1.9%. After 24 hours culture, sperm motility was 23.5$\pm$2.1% (Group I), 8.1$\pm$1.1% (Group II), 10.4$\pm$ 1.4% (Group III) and 4.0$\pm$0.8% (Group IV), respectively. After 48 hours, the motility had been changed as 32$\pm$2.3% (Group I), 14.3$\pm$1.7% (Group II), 5.3 $\pm$1.4% (Group III) and 4.3$\pm$0.9% (Group IV). In hFF group (I and II), sperm motility of group I cultured at 37$^{\circ}C$ was higher than those of group II at 32$^{\circ}C$. But, sperm viability of group I cultured at 37$^{\circ}C$ was lower than those of group II at 32$^{\circ}C$ (54.4$\pm$4.1% vs. 59.4$\pm$3.7%) after cultured for 48 hours. We acquired the best motility of testicular sperm when performed in vitro culture for 48 hours in hFF supplemented medium at 37$^{\circ}C$. Increase of sperm motility by in vitro culture could be useful tool fur human TESE-ICSI program.

• The Korean Journal of Food And Nutrition
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• v.10 no.2
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• pp.174-179
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• 1997

The antimicrobial and GTase(Glucosyltransferase) inhibition activity were investigated for solvent fractions of Gall-nut, variety of fork drugs and Red-grape husk water extracts. Among them, Gall-nut and Red-grape husk water extracts were selected for the powerful antimicrobial and GTase inhibition activity. The methanol fractions of Gall-nut and Red-grape husk were showed very powerful antimicrobial activity on both B. subtilis and E. coli. The MIC(Minimum Inhibitory Concentration) of gall-nut methanol fraction were 1.0mg/ml for B. subtilis and 3.0mg/ml for E. coli. Red-grape husk were 2.0mg/ml for B. subtilis and 3.0mg/ml for E. coli. The methanol fractions of Gall-nut and Red-grape husk were showed very powerful Gtase inhibition activity. The concentrations of these fractions for 80% inhibition of GTase activity were 1.08$\times$10-3mg/ml and 1.08$\times$10-2mg/ml, respectively. The principal compound for the antimicrobial and GTase inhibition activity in tese extracts seems to be polyphenol derivatives.

• Journal of Veterinary Clinics
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• v.17 no.2
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• pp.480-484
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• 2000

A 10 years old female Poodle weighing 4.38 kg and a 10 years old female Maltese weighing 4.33kg were referred to veterinary leaching hospital of college of veterinary medicine, Konkuk uni-versity because of hematuria and severa anorexia. Preoperative hematological and srum chemical values revealed that counts of WBC were $78.4{\times}10^3$,/TEX>/ul in the Poodle and $21.6{\times}10^3$in the Maltese, concentrations of BUN 105 and 13 mg/dl, concentrations of creatinine 4.0 and 1.4 mg/dl, concentration of ALKP 542 U/L in the Poodle and concentration of ALF 37 U/L in the Maltese, respectively. Radiographic findings were observed radiopaque bilateral nephroliths in the Poodle and right renal cal- culus in the Maltese. Ultrasonographic findings were showed hyperechoic mass with severe acoustic shadowing, which were ]ocated in the center of both kidneys parenchyma in the Poodle and in right kidney in the Maltese. Bisectional nephrotomy in right kidney for removal of calculus, which had ,sin- gle stone and more large than splitted stones in lets kidney in the Poodle, was performed under isnf- lurane anesthesia in 100% $O_2$. The time from temporary occlusion of blood flow to suture of incisioned kidney after removal of renal calculi was elapsed 12 minutes. During operation and until 24hrs after operation was infusioned mannitol and tranexanic acid solution. There were normal urination with yellow color in the Poodle at 3 days after operation and in the Maltese at 1 day and good appetite after 7 days and 1 day after operation. At 7 days and 9 days after operation, hematolgoical and seam chemical values repealed that counts of WBC were $29.8{\times}10^3/u1 in the Poodle and 13.1 {\itmes}10^3$/ul in the Mal- tese, concentrations of BUN 13 and 10 mg/dl. concentrations of creatinine 1.3 and 1.1 mg/dl, con-centrations of ALT 34 and 97U/L, concentraion of AST 10 U/L in the Poodle, respectively, By ultra-sonographic view there was not observed hyperechoic mass and radippaque calculus in operated kid-ney in radiographic view.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.3
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• pp.267-273
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• 2000

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 a computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from $LN_2$ and letting them at room temperature for 10 min followed by %37^{\circ}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 azoospennic 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.