- Email: [email protected]
Does severe teratozoospermia is correlated with embryonic aneuploidy rates?
with both euploid per biopsy rate and sperm concentration; however, there is no effect on other pregnancy parameters. The rate of miscarriage after IVFCCS with SET appears to be lower in couples with male factor infertility as compared to those with non-male factor etiologies, however this may be related to the incidence of PCOS and other etiologies of infertility in the non-male factor patients. Overall, the utility of IVF-CCS is a positive driver for cycle success in couples faced with male factor infertility. References: 1. Gianaroli L, Magli MC, Ferraretti AP. Sperm and blastomere aneuploidy detection in reproductive genetics and medicine. J Histochem Cytochem 2005; 53:261-67. 2. Coates A, et al. Use of suboptimal sperm increases the risk of aneuploidy of the sex chromosomes in preimplantation blastocyst embryos. Fertil Steril 2015; 104(4):866-72.
P-321 Tuesday, October 18, 2016 DETECTING Y-CHROMOSOME MICRODELETIONS USING NEXT GENERATION SEQUENCING (NGS) DATA. R. Shraga,a M. C. Akana,b S. L. Bristow,a A. Manoharan,a O. Puig.a aRecombine, New York, NY; bLab, Recombine, New York City, NY. OBJECTIVE: Male factor is at least partly responsible for infertility in approximately 50% of couples. Microdeletions in the Y-chromosome are identified in 5 - 13% of men with otherwise unexplained infertility.1 However, detection of microdeletions can be both complicated and expensive, and testing is not routinely performed. Our objective was to develop an assay that allows for detection of Y-chromosome microdeletions via Next Generation Sequencing (NGS). DESIGN: We designed a sequencing capture system with probes in the Y-chromosome AZFa and AZFbc regions, covering the exonic segments of all genes in the two regions. Samples were sequenced using Illumina NextSeq 500 and reads were mapped to the human genome using standard GATK pipeline. MATERIALS AND METHODS: We computed average depth per sample per interval and ran several normalization procedures to remove variation in depth due to non-biological noise. This procedure corrected for sample variability, batch effects, bias in GC content in the sequences, and other technical biases. After normalization, a copy number estimate per interval was computed by comparing each sample’s normalized depth per interval to the median normalized depth. Deletions were then called by running the Circular Binary Segmentation algorithm on the interval estimates. RESULTS: DNA samples from the Coriell repository were sequenced at different depths (100-300X). We detected unambiguously Y-chromosome microdeletions in all index cases. Deletions ranged in size from 2.3 to 7.7 Mbp. Data modeling indicates that microdeletions are detected confidently with the average sequencing depth of 100X, which is routinely used in clinical sequencing as recommended by the American College of Medical Genetics and Genomics guidelines. Further validation of the algorithm with additional clinical samples is ongoing. CONCLUSIONS: These results demonstrate that NGS is an inexpensive, accurate, and comprehensive method to detect Y-chromosome microdeletions. The advantage of NGS is that multiple mutations and chromosomal abnormalities can be screened in a single assay, enabling simultaneous analysis of various contributions to male infertility. Identifying the genetic basis for male infertility can guide decision-making around treatment, such as surgical intervention, use of ICSI, or use of donor sperm. Reference: 1. Silber SJ, Disteche CM. Y Chromosome Infertility. 2002 Oct 31 [Updated 2012 Oct 18]. In: Pagon RA, Adam MP, Ardinger HH, et al., editors. GeneReviewsÒ [Internet]. Seattle (WA): University of Washington, Seattle; 1993-2016. Available from: http://www.ncbi. nlm.nih.gov/books/NBK1339/.
P-322 Tuesday, October 18, 2016 EPIGENOME ANALYSIS USING INFINIUM 450K BEADCHIP ARRAYS IDENTIFIES ABERRANT DNA METHYLATION IN SPERMATOZOA OF MALES WITH UNEXPLAINED INFERTILITY. M. M. Laqqan,a Y. A. Alkhaled,a S. Tierling,b J. Walter,b M. E. Hammadeh.a aObstetrics & Gynecology, Saarland University, Germany, Homburg, Germany; bGenetik/Epigenetik, Saarland University, Germany, Saarbruecken, Germany.
FERTILITY & STERILITYÒ
OBJECTIVE: To determine whether DNA methylation at CpG dinucleotides is different in sperm DNA of subfertile and fertile male. DESIGN: Human semen samples were collected during the period between August 2014 to May 2015 from males who developed idiopathic infertility MATERIALS AND METHODS: Infinium 450K BeadChip arrays were used to identify genomic regions that show differences in sperm DNA methylation patterns between subfertile and fertile males from couples that have idiopathic infertility and underwent intracytoplasmic sperm injection. RESULTS: In this study 22 CpG dinucleotides have been found to be significantly and consistently different (between 19.33% and 86.46%, fdr-adj.p < 0.155) in the DNA methylation level in sperm samples of 3 subfertile and 3 fertile males. Only 7 of those (DNA methylation difference between 19.33% and 78.58%) do not overlap common annotated SNPs, 4 CpGs were found to be directly linked to the genes UBE2G2, ALDH3B2, PTGIR, ADAMTS14, 3 CpGs were located in intergenic DNAseI clusters. Currently, these results are under validation through local deep bisulfite sequencing and are planned to be tested on a larger sample cohort. CONCLUSIONS: The present study identified 7 consistently altered CpG dinucleotides in sperm, which may represent novel candidates related to idiopathic infertility. If detectable in a larger sample cohort these CpG sites might serve as a future diagnosing tool for male infertility. Supported by: Department of Obstetrics & Gynecology, University of Saarland. P-323 Tuesday, October 18, 2016 DOES SEVERE TERATOZOOSPERMIA IS CORRELATED WITH EMBRYONIC ANEUPLOIDY RATES?. J. Rodriguez-Purata,a L. Sekhon,a,b J. A. Lee,a M. C. Whitehouse,a R. Slifkin,a E. Flisser,a M. Duke,a A. B. Copperman,c,b B. Sandler,a,b N. Bar-Chama.a,b aReproductive Medicine Associates of New York, New York, NY; bObstetrics, Gynecology and Reproductive Science, Icahn School of Medicine at Mount Sinai, New York, NY; cObstetrics and Gynecology, RMANY-Mount Sinai, New York, NY. OBJECTIVE: Sperm volume, motility, and morphology are generally accepted as predictors of in vivo and in vitro fertilization. Kruger et al. demonstrated that microscopic assessment of morphology plays an integral role in evaluating a male patient. This study aims to determine if extremely low percentages of structurally normal sperm is correlated with increased rates of embryonic aneuploidy in couples who utilize IVF with preimplantation genetic screening (PGS). DESIGN: Retrospective. MATERIALS AND METHODS: Couples who underwent an IVF cycle and PGS from July 2010 - October 2015 were included. Sperm were assessed by Kruger’s strict criteria. Proportion of Kruger morphology was analyzed by male age (A: %35; B: (35-38]; C: (38-41]; D: (41-43]; and E: >43). Cohorts were segregated into two groups: Kruger %1%; and Kruger >4%. Couples were binned by female ages (A: %35; B: (3538]; C: (38-41]; D: (41-43]; and E: >43). Aneuploidy rate for each female age group and per study group was calculated, with 95% confidence intervals calculated by Clopper-Pearson method. Chi-square was used to test significance, established at p<0.05. RESULTS: Couples consisted of females (21.4-47.0 years) with male partners (22.6-70.3 years) who underwent 1105 autologous fresh IVF cycles with PGS. Trophectoderm biopsy was performed on 7927 embryos, of which 38.8% (n¼3077) were found aneuploid. The percentage of male patients with a morphology count %1% increased with male age (A: 10.4%, B: 12.4%, C: 26.0%, D: 21.9%, E: 37.2%; p<0.05). Fertilization rate was similar between group in each female age group. Aneuploidy rate was higher in couples where male Kruger morphology was %1% in each female age group, although it did not reach statistical significance (Table 1). CONCLUSIONS: Multiple studies have shown semen samples with poor Kruger morphology have similar fertilization and pregnancy rates to normal morphology when IVF/ICSI is utilized. In the present study, no correlation was identified between teratozoospermic specimens and increased incidence of embryonic aneuploidy. Male partners with specimens found to have severe teratozoospermia can be reassured that they do not have an increased incidence of producing chromosomally abnormal embryos. Further large randomize control trials are needed to confirm these findings.
e227
A: <35
Patient’s Age Partner’s Age BMI AMH Retrieved MII Fertilization Rate Fertilization Rate Aneuploidy rate
B: 35-38
C: 38-41
D: 41-43
E: >43
%1% (n¼101) // >4% (n¼235) 31.43.0 // 31.62.7, NS
%1% (n¼72) // >4% %1% (n¼83) // >4% %1% (n¼45) // >4% %1% (n¼25) // >4% (n¼163) (n¼231) (n¼109) (n¼41) 36.60.7 // 36.50.8, NS 39.50.8 // 39.50.8, NS 41.90.3 // 42.00.5, NS 43.91.1 // 43.80.8, NS
38.07.9 // 35.35.1, p<0.05 23.53.9 // 22.75.1, NS 3.12.1 // 3.83.2, NS 17.711.8 // 17.410.0, NS 14.39.2 // 13.98.5, NS 11.67.5 // 11.57.1, NS
39.25.7 // 38.84.1, NS 42.04.3 // 41.44.3, NS 43.64.1 // 43.44.4, NS 45.33.7 // 44.25.1, NS 23.75.0 // 22.94.0, NS 3.52.6 // 3.02.6, NS 13.49.4 // 16.99.3, NS 11.76.5 // 12.77.4, NS 9.15.8 // 10.67.1, NS
25.65.6 // 23.314.1, NS 2.42.1 // 2.62.3, NS 13.47.8 // 13.17.9, NS 9.55.7 // 9.76.1, NS 7.75.1 // 7.65.2, NS
25.54.0 // 23.24.4, NS 1.00.7 // 1.51.1, NS 12.38.6 // 12.17.2, NS 9.27.6 // 9.45.8, NS 6.25.5 // 7.04.5, NS
25.63.1 // 23.43.5, NS 1.20.8 // 1.51.2, NS 16.27.4 // 11.77.9, NS 9.97.4 // 8.36.4, NS 6.85.5 // 6.95.4, NS
81.0% (1171/1445) // 82.2% 79.8% (657/823) // 81.7% 76.4% (603/789) // 78.6% 6.25.5 // 7.04.5, NS 69.2% (171/247) // 67.1% (2692/3275) , NS (1730/2117), NS (1768/2249), NS (202/301), NS 26.0% (168/645) // 26.6% 36.9% (114/195) // 31.1% 54.2% (166/306) // 49.9% 54.7% (133/243) // 51.7% 65.7% (44/67) // 60.0% (404/1520), NS (308/991), NS (402/805), NS (186/360), NS (69/115), NS
P-324 Tuesday, October 18, 2016
SUCCESSFUL TREATMENTS FOR KARTAGENER SYNDROME WITH COMPLETELY IMMOTILE SPERM OR AZOOSPERMIA. A. Tanaka, M. Nagayoshi, I. Tanaka, T. Miki, T. Yamaguchi. Saint Mother Hospital, Kitakyusyu, Japan. OBJECTIVE: Kartagener triad syndrome consists of dextrocardia, bronchoectasia and chronic bronchitis. It is often associated with completely immotile sperm with structurally abnormal tails. However, we observed that about half of Kartagener syndrome cases show obstructive azoospermia with normal epididymal spermatozoa with structurally normal flagellum collected easily with MESA (Microsurgical Epididymal Sperm Aspiration). We report successful cases of Kartagener syndrome with completely immotile sperm or obstructive azoospermia. DESIGN: Retrospective case series. MATERIALS AND METHODS: Viability of completely immotile sperms was investigated with eosin-negrosin test. We conducted HOST to choose the viable spermatozoon when the positive ratio was over 30%. This test is based on the principle that viable spermatozoa have intact membranes which shows swelling of the cytoplasmic space and curling of the sperm tail. The spermatozoon with HOST positive was available for use in ICSI. In the case of lower positive ratio (<30%), MESA was conducted. Scrototomy is usually performed under general (Propofol + Fentanyl citrate) or loco-regional (cord block) anesthesia
(10ml of 1% Lidocaine + Anapeine in equal quantities). High quality sperm is retrieved from whitish regions and low quality sperm from yellowish regions at the head of epididymis through insertion of a glass pipette with a sharp angle. It is easy to collect a large volume of sperm from the white regions. RESULTS: We summarized clinical outcome of Kartagener syndrome in Table. CONCLUSIONS: Kartagener syndrome is now regarded as one type of primary ciliary dyskinesia (PCD) which is accompanied with ultrastructural or functional defects of cilia (flagellum) and higher probability of conception by selecting the available spermatozoon. P-325 Tuesday, October 18, 2016 OUTCOMES OF ICSI CYCLES WITH EITHER FRESH OR FROZEN-THAWED TESTICULAR SPERMATOZOA IN CONSECUTIVE CYCLES OF NON-OBSTRUCTIVE AZOOSPERMIC PATIENTS. U. Ozdemir,a E. E. Nal,b H. K. Yelke,a C. Pirkevi Cetinkaya,a M. M. Basar,a S. Kahraman.a aAssisted Reproductive Technologies and Reproductive Genetics Center, Istanbul Memorial Hospital, Istanbul, Turkey; bMemorial Sisli Hospital, Istanbul, Turkey. OBJECTIVE: To compare pregnancy outcomes of first ICSI cycles with fresh testicular spermatozoa in non-obstructive azoospermic patients with consecutive cycles, in whom frozen-thawed testicular spermatozoa were used one to three times during study period.
Successful treatments for Kartagener syndrome with immotile sperm or azoospermia (*: Brothers)
Viability with Sperm eosin-negrosin Count test (%) / HOST (x100000) / (positive, Motility (%) / 2PN negative) FSH (mIU/ml) fertilization (%) Embryo transfer (n) Immotile ejaculate sperms Obstructive azoospermia
e228
Clinical outcome
TEM of flagellum
1 * 65 / positive
500 / 0 / 5.4
66.7 (8/12)
Blastocyst 5AA(1)
Relivery _ (17y, normal sperm) Abnormal
2 * 55 / positive 3 45 / positive 4 Motile
500 / 0 / 6.3 100 / 0 / 7.5 250 / 40 / 6.7
70.0 (7/10) 42.9 (3/7) 66.7 (4/6)
Blastocyst 4AA(1) Blastocyst 1BA(1) Blastocyst 4AA(1)
Relivery _ (14y) No pregnancy Relivery \ (10y)
Abnormal Abnormal Normal
5 6
250 / 40 / 6.7 50 / 70 / 6.5
75.0 (3/4) 50.0 (4/8)
Blastocyst 3AA(1) Blastocyst 1AA(1)
Relivery \ (6y) No pregnancy
Normal Normal
Motile Motile
ASRM Abstracts
Vol. 106, No. 3, Supplement, September 2016
P-321 Tuesday, October 18, 2016 DETECTING Y-CHROMOSOME MICRODELETIONS USING NEXT GENERATION SEQUENCING (NGS) DATA. R. Shraga,a M. C. Akana,b S. L. Bristow,a A. Manoharan,a O. Puig.a aRecombine, New York, NY; bLab, Recombine, New York City, NY. OBJECTIVE: Male factor is at least partly responsible for infertility in approximately 50% of couples. Microdeletions in the Y-chromosome are identified in 5 - 13% of men with otherwise unexplained infertility.1 However, detection of microdeletions can be both complicated and expensive, and testing is not routinely performed. Our objective was to develop an assay that allows for detection of Y-chromosome microdeletions via Next Generation Sequencing (NGS). DESIGN: We designed a sequencing capture system with probes in the Y-chromosome AZFa and AZFbc regions, covering the exonic segments of all genes in the two regions. Samples were sequenced using Illumina NextSeq 500 and reads were mapped to the human genome using standard GATK pipeline. MATERIALS AND METHODS: We computed average depth per sample per interval and ran several normalization procedures to remove variation in depth due to non-biological noise. This procedure corrected for sample variability, batch effects, bias in GC content in the sequences, and other technical biases. After normalization, a copy number estimate per interval was computed by comparing each sample’s normalized depth per interval to the median normalized depth. Deletions were then called by running the Circular Binary Segmentation algorithm on the interval estimates. RESULTS: DNA samples from the Coriell repository were sequenced at different depths (100-300X). We detected unambiguously Y-chromosome microdeletions in all index cases. Deletions ranged in size from 2.3 to 7.7 Mbp. Data modeling indicates that microdeletions are detected confidently with the average sequencing depth of 100X, which is routinely used in clinical sequencing as recommended by the American College of Medical Genetics and Genomics guidelines. Further validation of the algorithm with additional clinical samples is ongoing. CONCLUSIONS: These results demonstrate that NGS is an inexpensive, accurate, and comprehensive method to detect Y-chromosome microdeletions. The advantage of NGS is that multiple mutations and chromosomal abnormalities can be screened in a single assay, enabling simultaneous analysis of various contributions to male infertility. Identifying the genetic basis for male infertility can guide decision-making around treatment, such as surgical intervention, use of ICSI, or use of donor sperm. Reference: 1. Silber SJ, Disteche CM. Y Chromosome Infertility. 2002 Oct 31 [Updated 2012 Oct 18]. In: Pagon RA, Adam MP, Ardinger HH, et al., editors. GeneReviewsÒ [Internet]. Seattle (WA): University of Washington, Seattle; 1993-2016. Available from: http://www.ncbi. nlm.nih.gov/books/NBK1339/.
P-322 Tuesday, October 18, 2016 EPIGENOME ANALYSIS USING INFINIUM 450K BEADCHIP ARRAYS IDENTIFIES ABERRANT DNA METHYLATION IN SPERMATOZOA OF MALES WITH UNEXPLAINED INFERTILITY. M. M. Laqqan,a Y. A. Alkhaled,a S. Tierling,b J. Walter,b M. E. Hammadeh.a aObstetrics & Gynecology, Saarland University, Germany, Homburg, Germany; bGenetik/Epigenetik, Saarland University, Germany, Saarbruecken, Germany.
FERTILITY & STERILITYÒ
OBJECTIVE: To determine whether DNA methylation at CpG dinucleotides is different in sperm DNA of subfertile and fertile male. DESIGN: Human semen samples were collected during the period between August 2014 to May 2015 from males who developed idiopathic infertility MATERIALS AND METHODS: Infinium 450K BeadChip arrays were used to identify genomic regions that show differences in sperm DNA methylation patterns between subfertile and fertile males from couples that have idiopathic infertility and underwent intracytoplasmic sperm injection. RESULTS: In this study 22 CpG dinucleotides have been found to be significantly and consistently different (between 19.33% and 86.46%, fdr-adj.p < 0.155) in the DNA methylation level in sperm samples of 3 subfertile and 3 fertile males. Only 7 of those (DNA methylation difference between 19.33% and 78.58%) do not overlap common annotated SNPs, 4 CpGs were found to be directly linked to the genes UBE2G2, ALDH3B2, PTGIR, ADAMTS14, 3 CpGs were located in intergenic DNAseI clusters. Currently, these results are under validation through local deep bisulfite sequencing and are planned to be tested on a larger sample cohort. CONCLUSIONS: The present study identified 7 consistently altered CpG dinucleotides in sperm, which may represent novel candidates related to idiopathic infertility. If detectable in a larger sample cohort these CpG sites might serve as a future diagnosing tool for male infertility. Supported by: Department of Obstetrics & Gynecology, University of Saarland. P-323 Tuesday, October 18, 2016 DOES SEVERE TERATOZOOSPERMIA IS CORRELATED WITH EMBRYONIC ANEUPLOIDY RATES?. J. Rodriguez-Purata,a L. Sekhon,a,b J. A. Lee,a M. C. Whitehouse,a R. Slifkin,a E. Flisser,a M. Duke,a A. B. Copperman,c,b B. Sandler,a,b N. Bar-Chama.a,b aReproductive Medicine Associates of New York, New York, NY; bObstetrics, Gynecology and Reproductive Science, Icahn School of Medicine at Mount Sinai, New York, NY; cObstetrics and Gynecology, RMANY-Mount Sinai, New York, NY. OBJECTIVE: Sperm volume, motility, and morphology are generally accepted as predictors of in vivo and in vitro fertilization. Kruger et al. demonstrated that microscopic assessment of morphology plays an integral role in evaluating a male patient. This study aims to determine if extremely low percentages of structurally normal sperm is correlated with increased rates of embryonic aneuploidy in couples who utilize IVF with preimplantation genetic screening (PGS). DESIGN: Retrospective. MATERIALS AND METHODS: Couples who underwent an IVF cycle and PGS from July 2010 - October 2015 were included. Sperm were assessed by Kruger’s strict criteria. Proportion of Kruger morphology was analyzed by male age (A: %35; B: (35-38]; C: (38-41]; D: (41-43]; and E: >43). Cohorts were segregated into two groups: Kruger %1%; and Kruger >4%. Couples were binned by female ages (A: %35; B: (3538]; C: (38-41]; D: (41-43]; and E: >43). Aneuploidy rate for each female age group and per study group was calculated, with 95% confidence intervals calculated by Clopper-Pearson method. Chi-square was used to test significance, established at p<0.05. RESULTS: Couples consisted of females (21.4-47.0 years) with male partners (22.6-70.3 years) who underwent 1105 autologous fresh IVF cycles with PGS. Trophectoderm biopsy was performed on 7927 embryos, of which 38.8% (n¼3077) were found aneuploid. The percentage of male patients with a morphology count %1% increased with male age (A: 10.4%, B: 12.4%, C: 26.0%, D: 21.9%, E: 37.2%; p<0.05). Fertilization rate was similar between group in each female age group. Aneuploidy rate was higher in couples where male Kruger morphology was %1% in each female age group, although it did not reach statistical significance (Table 1). CONCLUSIONS: Multiple studies have shown semen samples with poor Kruger morphology have similar fertilization and pregnancy rates to normal morphology when IVF/ICSI is utilized. In the present study, no correlation was identified between teratozoospermic specimens and increased incidence of embryonic aneuploidy. Male partners with specimens found to have severe teratozoospermia can be reassured that they do not have an increased incidence of producing chromosomally abnormal embryos. Further large randomize control trials are needed to confirm these findings.
e227
A: <35
Patient’s Age Partner’s Age BMI AMH Retrieved MII Fertilization Rate Fertilization Rate Aneuploidy rate
B: 35-38
C: 38-41
D: 41-43
E: >43
%1% (n¼101) // >4% (n¼235) 31.43.0 // 31.62.7, NS
%1% (n¼72) // >4% %1% (n¼83) // >4% %1% (n¼45) // >4% %1% (n¼25) // >4% (n¼163) (n¼231) (n¼109) (n¼41) 36.60.7 // 36.50.8, NS 39.50.8 // 39.50.8, NS 41.90.3 // 42.00.5, NS 43.91.1 // 43.80.8, NS
38.07.9 // 35.35.1, p<0.05 23.53.9 // 22.75.1, NS 3.12.1 // 3.83.2, NS 17.711.8 // 17.410.0, NS 14.39.2 // 13.98.5, NS 11.67.5 // 11.57.1, NS
39.25.7 // 38.84.1, NS 42.04.3 // 41.44.3, NS 43.64.1 // 43.44.4, NS 45.33.7 // 44.25.1, NS 23.75.0 // 22.94.0, NS 3.52.6 // 3.02.6, NS 13.49.4 // 16.99.3, NS 11.76.5 // 12.77.4, NS 9.15.8 // 10.67.1, NS
25.65.6 // 23.314.1, NS 2.42.1 // 2.62.3, NS 13.47.8 // 13.17.9, NS 9.55.7 // 9.76.1, NS 7.75.1 // 7.65.2, NS
25.54.0 // 23.24.4, NS 1.00.7 // 1.51.1, NS 12.38.6 // 12.17.2, NS 9.27.6 // 9.45.8, NS 6.25.5 // 7.04.5, NS
25.63.1 // 23.43.5, NS 1.20.8 // 1.51.2, NS 16.27.4 // 11.77.9, NS 9.97.4 // 8.36.4, NS 6.85.5 // 6.95.4, NS
81.0% (1171/1445) // 82.2% 79.8% (657/823) // 81.7% 76.4% (603/789) // 78.6% 6.25.5 // 7.04.5, NS 69.2% (171/247) // 67.1% (2692/3275) , NS (1730/2117), NS (1768/2249), NS (202/301), NS 26.0% (168/645) // 26.6% 36.9% (114/195) // 31.1% 54.2% (166/306) // 49.9% 54.7% (133/243) // 51.7% 65.7% (44/67) // 60.0% (404/1520), NS (308/991), NS (402/805), NS (186/360), NS (69/115), NS
P-324 Tuesday, October 18, 2016
SUCCESSFUL TREATMENTS FOR KARTAGENER SYNDROME WITH COMPLETELY IMMOTILE SPERM OR AZOOSPERMIA. A. Tanaka, M. Nagayoshi, I. Tanaka, T. Miki, T. Yamaguchi. Saint Mother Hospital, Kitakyusyu, Japan. OBJECTIVE: Kartagener triad syndrome consists of dextrocardia, bronchoectasia and chronic bronchitis. It is often associated with completely immotile sperm with structurally abnormal tails. However, we observed that about half of Kartagener syndrome cases show obstructive azoospermia with normal epididymal spermatozoa with structurally normal flagellum collected easily with MESA (Microsurgical Epididymal Sperm Aspiration). We report successful cases of Kartagener syndrome with completely immotile sperm or obstructive azoospermia. DESIGN: Retrospective case series. MATERIALS AND METHODS: Viability of completely immotile sperms was investigated with eosin-negrosin test. We conducted HOST to choose the viable spermatozoon when the positive ratio was over 30%. This test is based on the principle that viable spermatozoa have intact membranes which shows swelling of the cytoplasmic space and curling of the sperm tail. The spermatozoon with HOST positive was available for use in ICSI. In the case of lower positive ratio (<30%), MESA was conducted. Scrototomy is usually performed under general (Propofol + Fentanyl citrate) or loco-regional (cord block) anesthesia
(10ml of 1% Lidocaine + Anapeine in equal quantities). High quality sperm is retrieved from whitish regions and low quality sperm from yellowish regions at the head of epididymis through insertion of a glass pipette with a sharp angle. It is easy to collect a large volume of sperm from the white regions. RESULTS: We summarized clinical outcome of Kartagener syndrome in Table. CONCLUSIONS: Kartagener syndrome is now regarded as one type of primary ciliary dyskinesia (PCD) which is accompanied with ultrastructural or functional defects of cilia (flagellum) and higher probability of conception by selecting the available spermatozoon. P-325 Tuesday, October 18, 2016 OUTCOMES OF ICSI CYCLES WITH EITHER FRESH OR FROZEN-THAWED TESTICULAR SPERMATOZOA IN CONSECUTIVE CYCLES OF NON-OBSTRUCTIVE AZOOSPERMIC PATIENTS. U. Ozdemir,a E. E. Nal,b H. K. Yelke,a C. Pirkevi Cetinkaya,a M. M. Basar,a S. Kahraman.a aAssisted Reproductive Technologies and Reproductive Genetics Center, Istanbul Memorial Hospital, Istanbul, Turkey; bMemorial Sisli Hospital, Istanbul, Turkey. OBJECTIVE: To compare pregnancy outcomes of first ICSI cycles with fresh testicular spermatozoa in non-obstructive azoospermic patients with consecutive cycles, in whom frozen-thawed testicular spermatozoa were used one to three times during study period.
Successful treatments for Kartagener syndrome with immotile sperm or azoospermia (*: Brothers)
Viability with Sperm eosin-negrosin Count test (%) / HOST (x100000) / (positive, Motility (%) / 2PN negative) FSH (mIU/ml) fertilization (%) Embryo transfer (n) Immotile ejaculate sperms Obstructive azoospermia
e228
Clinical outcome
TEM of flagellum
1 * 65 / positive
500 / 0 / 5.4
66.7 (8/12)
Blastocyst 5AA(1)
Relivery _ (17y, normal sperm) Abnormal
2 * 55 / positive 3 45 / positive 4 Motile
500 / 0 / 6.3 100 / 0 / 7.5 250 / 40 / 6.7
70.0 (7/10) 42.9 (3/7) 66.7 (4/6)
Blastocyst 4AA(1) Blastocyst 1BA(1) Blastocyst 4AA(1)
Relivery _ (14y) No pregnancy Relivery \ (10y)
Abnormal Abnormal Normal
5 6
250 / 40 / 6.7 50 / 70 / 6.5
75.0 (3/4) 50.0 (4/8)
Blastocyst 3AA(1) Blastocyst 1AA(1)
Relivery \ (6y) No pregnancy
Normal Normal
Motile Motile
ASRM Abstracts
Vol. 106, No. 3, Supplement, September 2016