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Association between body mass index (BMI) and semen quality
program defined HA swimming patterns: thrash, star, helical, & circling, or all four together. HDATA pulled data from each analysis into Excel files for later HA sorting according to 12 equations. MATERIALS AND METHODS: Over 50,000 sperm were evaluated, from 9 different donors. Sperm were sampled at all stages of CAP. 1) % of sperm with the individual HA patterns, or ALL HA, when Burkman HA criteria were put into SORT; 2) and when the sperm were reassessed using M&M 1990 criteria; 3) The same comparison was performed, but using the 1995 criteria; 4) Modifications to the 1995 criteria were tested, especially amplitude. Statistics: regression, ANOVA, chi-squared. RESULTS: Readings of human sperm using the Burkman 1990 criteria were compared to the M&M 1990 definitions: no statistical difference (P equal to 0.47). Comparison of Burkman criteria for HA to M&M 1995 values gave a P value less than 0.001. Parallel comparisons were made for individual patterns. Burkman values for Helical, Thrash and Star, added together, were virtually equal to the 1995 result. Inclusion of Circling dropped dramatically with M&M 1995. CONCLUSION: Pregnancy, as an endpoint, is our gold standard (see other abstract). We find no Pregnancy data as validation for the 1995 paper. LifeCell Dx can provide guidelines for initiating Clear CASA in any lab. All labs must validate their own specific protocols for Clear CASA (medium, temperature, sperm handling, etc.). We emphasize that Circling HA adheres o the classical drawings of HA.
catu Medical School -Sao Paulo State University - UNESP, Botucatu, Sao Paulo, Brazil. OBJECTIVE: To analyze whether there are correlations between the LIF thymine (T)/guanine (G) (rs929271) polymorphism and etiopathogenic factors of DNA damage in sperm. DESIGN: Cross-sectional. MATERIALS AND METHODS: A total of 177 men were included. DNA was extracted from peripheral blood, and LIF gene single-nucleotide polymorphisms (SNPs) T/G (rs929271) were genotyped using real-time PCR with Taqman Universal PCR Master Mix and Taqman SNP genotyping assays. At least 200 spermatozoa were examined in each semen sample (one per subject) and the following spermatozoa percentages were determined: DNA fragmentation (with TUNEL assay), abnormal chromatin packaging/ underprotamination (using chromomycin A3/CMA3), apoptosis (using annexin-V) and mitochondrial damage (using MitoTracker Green and JC1). All data were evaluated using c2 tests. RESULTS: The LIF genotypes with G alleles were associated with high percentages of spermatozoa with abnormal chromatin packaging. There was no observed association between genotypes or alleles of LIF and DNA fragmentation, apoptosis or mitochondrial damage in sperm (Table 1). TABLE 1. %Mitochondrial damage
P-994 Thursday, October 17, 2013 ASSOCIATION BETWEEN BODY MASS INDEX (BMI) AND SEMEN QUALITY. J. K. Rodrigues, F. S. Vieira, C. C. Moraes, M. H. Furtado, A. M. M. Cota, J. P. J. Caetano. Department of Research and Development, Pr o-Criar Reproductive Medicine - Brazil, Belo Horizonte, Minas Gerais, Brazil. OBJECTIVE: The aim of this study was to explore the correlation between BMI and semen quality. DESIGN: Retrospective study. MATERIALS AND METHODS: Analysis of 45 male patients who presented for clinical evaluation in our service in 2012. Of the total surveyed, 17 presented normal BMI (BMI <25 kg / m2) and 28 patients were diagnosed with overweight (BMI R 25 kg/m2). Exclusion criteria were: previous treatment for infertility, use of drugs that can alter spermatogenesis, vasectomy, diagnosis of varicocele treated or not, cryptorchidism or clinical/surgical problems with risk for fertility, smoking and sexually transmitted diseases. The seminal parameters analyzed were: ejaculate volume, sperm concentration, percentage of progressive motile spermatozoa, morphology and BMI. For statistical analysis we used the Student’s t test followed by Mann-Whitney test. RESULTS: No difference was found between the analyzed parameters between the groups with BMI < 25 kg/m2 or BMI R 25 kg/m2, except BMI (22.4 vs. 28.7 - p <0.001). However there was a trend to decreased sperm concentration (55.3 M/mL vs. 26.4 M/mL - p ¼ 0.06) and increased ejaculate volume (2.9 mL vs. 4.2 mL - p ¼ 0.06) in patients with BMI R 25 kg/m2. The mean values of motility (53% vs. 40%) and morphology (4% vs. 3%) also are decreased in this group compared to the group with BMI <25 kg/m2. Furthermore, it was found that the group with BMI R 25 kg/m2, had a higher percentage of patients with semen parameters below the normal standards by the World Health Organization, compared to the group with BMI < 25 kg/ m2 (concentration < 15 M/m: 39% vs. 24%; motility < 32%: 39% vs. 12%; morphology < 4%: 73% vs. 53%). CONCLUSION: These results indicate that BMI may alter sperm quality, resulting in low sperm concentration, reduced motility and altered morphology. Suggest to increase the sample size to confirm these data. Supported by: Pr o-Criar Reproductive Medicine.
P-995 Thursday, October 17, 2013 RELATIONSHIP BETWEEN LEUKEMIA INHIBITORY FACTOR (LIF) POLYMORPHISMS AND ETIOPATHOGENIC FACTORS OF DNA DAMAGE IN SPERM. L. D. Vagnini,a C. G. Petersen,a,b,c A. L. Mauri,a,b J. B. A. Oliveira,a,b,c R. L. R. Baruffi,a,b J. G. Franco, Jr.a,b,c a Paulista Center for Diagnosis Research and Training, Ribeirao Preto, Sao Paulo, Brazil; bCenter for Human Reproduction Prof. Franco Jr, Ribeirao Preto, Sao Paulo, Brazil; cDepartment of Gynecology and Obstetrics, Botu-
FERTILITY & STERILITYÒ
LIF Genotypes T/T T/G G/G P Genotypes T/T T/G+G/G P Alleles T G P
%DNA fragmentation
%CMA3 positivity
%Apoptosis
MitoTracker Green
JC1
13.67.7 12.06.0 13.07.5 0.58
5117.2 19.46.2 57.815.3 20.36.8 58.515.1 18.45.8 0.03 0.67
29.915.7 27.719.7 27.621.2 0.62
34.618.3 33.422.2 33.412.5 0.73
13.67.7 12.16.2 0.31
51.017.2 19.46.2 57.915.2 206.6 0.01 0.83
29.915.7 27.719.8 0.33
34.618.3 33.420.6 0.56
12.86.9 12.16.2 0.60
54.516.5 19.96.5 57.915.2 206.6 0.15 0.98
28.717.9 27.719.8 0.58
33.920.4 33.420.6 0.82
CONCLUSION: The results indicate that LIF gene T/G polymorphisms are related to sperm protamination (worse results associated with the G allele). Further investigations should be conducted to confirm these findings and to determine their clinical implications.
P-996 Thursday, October 17, 2013 MIND THE GAP: SUBJECTS BELOW WORLD HEALTH ORGANIZATION 4TH ED (WHO 4) BUT ABOVE WHO 5TH ED (WHO 5) MINIMUM REFERENCE VALUES HAVE SUBSTANTIAL ODDS OF ABNORMAL SEMEN RESULTS ON SUBSEQUENT TESTING. K. Baker, Y. Barazani, E. Sabanegh. Center for Male Fertility, Cleveland Clinic, Cleveland, OH. OBJECTIVE: Numerous studies have established an overlap between semen parameters of fertile and infertile couples. The concern that couples may be excluded from fertility evaluation based on semen parameters above ‘‘normal’’ limits is justified because many men are screened by providers without dedicated male fertility training. We sought to categorize men seeking fertility evaluation based on WHO 4 and WHO 5 criteria and determine the odds of changing categories during serial testing. DESIGN: Retrospective chart review. MATERIALS AND METHODS: All male fertility referrals from 1/2007 through 12/2011 with at least 1 semen analysis at our facility were examined and diagnosis codes recorded. Subjects on medications that impact sperm production were excluded. Subjects were excluded from further analysis after varicocelectomy or if their initial results were normal by WHO 4. Results
S435
catu Medical School -Sao Paulo State University - UNESP, Botucatu, Sao Paulo, Brazil. OBJECTIVE: To analyze whether there are correlations between the LIF thymine (T)/guanine (G) (rs929271) polymorphism and etiopathogenic factors of DNA damage in sperm. DESIGN: Cross-sectional. MATERIALS AND METHODS: A total of 177 men were included. DNA was extracted from peripheral blood, and LIF gene single-nucleotide polymorphisms (SNPs) T/G (rs929271) were genotyped using real-time PCR with Taqman Universal PCR Master Mix and Taqman SNP genotyping assays. At least 200 spermatozoa were examined in each semen sample (one per subject) and the following spermatozoa percentages were determined: DNA fragmentation (with TUNEL assay), abnormal chromatin packaging/ underprotamination (using chromomycin A3/CMA3), apoptosis (using annexin-V) and mitochondrial damage (using MitoTracker Green and JC1). All data were evaluated using c2 tests. RESULTS: The LIF genotypes with G alleles were associated with high percentages of spermatozoa with abnormal chromatin packaging. There was no observed association between genotypes or alleles of LIF and DNA fragmentation, apoptosis or mitochondrial damage in sperm (Table 1). TABLE 1. %Mitochondrial damage
P-994 Thursday, October 17, 2013 ASSOCIATION BETWEEN BODY MASS INDEX (BMI) AND SEMEN QUALITY. J. K. Rodrigues, F. S. Vieira, C. C. Moraes, M. H. Furtado, A. M. M. Cota, J. P. J. Caetano. Department of Research and Development, Pr o-Criar Reproductive Medicine - Brazil, Belo Horizonte, Minas Gerais, Brazil. OBJECTIVE: The aim of this study was to explore the correlation between BMI and semen quality. DESIGN: Retrospective study. MATERIALS AND METHODS: Analysis of 45 male patients who presented for clinical evaluation in our service in 2012. Of the total surveyed, 17 presented normal BMI (BMI <25 kg / m2) and 28 patients were diagnosed with overweight (BMI R 25 kg/m2). Exclusion criteria were: previous treatment for infertility, use of drugs that can alter spermatogenesis, vasectomy, diagnosis of varicocele treated or not, cryptorchidism or clinical/surgical problems with risk for fertility, smoking and sexually transmitted diseases. The seminal parameters analyzed were: ejaculate volume, sperm concentration, percentage of progressive motile spermatozoa, morphology and BMI. For statistical analysis we used the Student’s t test followed by Mann-Whitney test. RESULTS: No difference was found between the analyzed parameters between the groups with BMI < 25 kg/m2 or BMI R 25 kg/m2, except BMI (22.4 vs. 28.7 - p <0.001). However there was a trend to decreased sperm concentration (55.3 M/mL vs. 26.4 M/mL - p ¼ 0.06) and increased ejaculate volume (2.9 mL vs. 4.2 mL - p ¼ 0.06) in patients with BMI R 25 kg/m2. The mean values of motility (53% vs. 40%) and morphology (4% vs. 3%) also are decreased in this group compared to the group with BMI <25 kg/m2. Furthermore, it was found that the group with BMI R 25 kg/m2, had a higher percentage of patients with semen parameters below the normal standards by the World Health Organization, compared to the group with BMI < 25 kg/ m2 (concentration < 15 M/m: 39% vs. 24%; motility < 32%: 39% vs. 12%; morphology < 4%: 73% vs. 53%). CONCLUSION: These results indicate that BMI may alter sperm quality, resulting in low sperm concentration, reduced motility and altered morphology. Suggest to increase the sample size to confirm these data. Supported by: Pr o-Criar Reproductive Medicine.
P-995 Thursday, October 17, 2013 RELATIONSHIP BETWEEN LEUKEMIA INHIBITORY FACTOR (LIF) POLYMORPHISMS AND ETIOPATHOGENIC FACTORS OF DNA DAMAGE IN SPERM. L. D. Vagnini,a C. G. Petersen,a,b,c A. L. Mauri,a,b J. B. A. Oliveira,a,b,c R. L. R. Baruffi,a,b J. G. Franco, Jr.a,b,c a Paulista Center for Diagnosis Research and Training, Ribeirao Preto, Sao Paulo, Brazil; bCenter for Human Reproduction Prof. Franco Jr, Ribeirao Preto, Sao Paulo, Brazil; cDepartment of Gynecology and Obstetrics, Botu-
FERTILITY & STERILITYÒ
LIF Genotypes T/T T/G G/G P Genotypes T/T T/G+G/G P Alleles T G P
%DNA fragmentation
%CMA3 positivity
%Apoptosis
MitoTracker Green
JC1
13.67.7 12.06.0 13.07.5 0.58
5117.2 19.46.2 57.815.3 20.36.8 58.515.1 18.45.8 0.03 0.67
29.915.7 27.719.7 27.621.2 0.62
34.618.3 33.422.2 33.412.5 0.73
13.67.7 12.16.2 0.31
51.017.2 19.46.2 57.915.2 206.6 0.01 0.83
29.915.7 27.719.8 0.33
34.618.3 33.420.6 0.56
12.86.9 12.16.2 0.60
54.516.5 19.96.5 57.915.2 206.6 0.15 0.98
28.717.9 27.719.8 0.58
33.920.4 33.420.6 0.82
CONCLUSION: The results indicate that LIF gene T/G polymorphisms are related to sperm protamination (worse results associated with the G allele). Further investigations should be conducted to confirm these findings and to determine their clinical implications.
P-996 Thursday, October 17, 2013 MIND THE GAP: SUBJECTS BELOW WORLD HEALTH ORGANIZATION 4TH ED (WHO 4) BUT ABOVE WHO 5TH ED (WHO 5) MINIMUM REFERENCE VALUES HAVE SUBSTANTIAL ODDS OF ABNORMAL SEMEN RESULTS ON SUBSEQUENT TESTING. K. Baker, Y. Barazani, E. Sabanegh. Center for Male Fertility, Cleveland Clinic, Cleveland, OH. OBJECTIVE: Numerous studies have established an overlap between semen parameters of fertile and infertile couples. The concern that couples may be excluded from fertility evaluation based on semen parameters above ‘‘normal’’ limits is justified because many men are screened by providers without dedicated male fertility training. We sought to categorize men seeking fertility evaluation based on WHO 4 and WHO 5 criteria and determine the odds of changing categories during serial testing. DESIGN: Retrospective chart review. MATERIALS AND METHODS: All male fertility referrals from 1/2007 through 12/2011 with at least 1 semen analysis at our facility were examined and diagnosis codes recorded. Subjects on medications that impact sperm production were excluded. Subjects were excluded from further analysis after varicocelectomy or if their initial results were normal by WHO 4. Results
S435