Pavithra Ranganathan

Creatine kinase as an indicator of sperm quality and maturity in men with oligospermia.

OBJECTIVES To determine the differences among the creatine kinase (CK) levels in the spermatozoa of subfertile men with mild, moderate, or severe oligospermia and to examine the differences in CK activity between infertile patients with various clinical diagnoses and a group of normal healthy donors (control). CK is a marker of sperm maturity that correlates with the sperm fertilizing capacity. Elevated levels are associated with an increased rate of functional abnormalities and increased cytoplasmic retention. METHODS We compared the CK levels in 51 oligospermic men who could not initiate a pregnancy. Patients were categorized according to their degree of oligospermia as defined by the total sperm count: mild (greater than 10 to 40 x 10(6); n = 30), moderate (5 to 10 x 10(6); n = 11), and severe (less than 5 x 10(6); n = 10). These patients were further classified according to their diagnosis (ie, varicocele, n = 24; unexplained infertility, n = 17; vasectomy reversal, n = 9; and unknown diagnosis, n = 1). A separate group consisting of 25 healthy donors was included as a control group. A computer-assisted semen analyzer assessed the sperm characteristics, and the CK levels were measured using a CK test kit after the enzyme was extracted with Triton-X. RESULTS The CK levels were significantly higher in the sperm of the severely oligospermic group (8.8 +/- 6.5 IU/10(8) sperm) than in the moderate (0.50 +/- 0.19 IU/10(8) sperm) and mild (0.49 +/- 0.15 IU/10(8) sperm) groups (P <0.0001). The mean CK level in the severely oligospermic group was 18-fold higher than that in the moderate (P = 0.03) and mild (P <0.001) groups. The CK levels were significantly higher in all three infertile groups compared with the donor group (0.06 +/- 0.01 IU/10(8) sperm). Patients with varicocele had the highest CK level (3.42 +/- 2.56 IU/10(8) sperm) compared with patients in the vasectomy reversal group (1.73 +/- 0.98 IU/10(8) sperm) and the idiopathic infertility group (0.26 +/- 0.08 IU/10(8) sperm). CONCLUSIONS Elevated CK levels are associated with severe oligospermia, irrespective of the clinical diagnosis. CK may be a sensitive indicator of sperm quality and maturity in the follow-up of patients treated for male factor infertility.

Comparison of two cryopreservation protocols for freezing human spermatozoa.

FREEZING HUMAN SPERMATOZOA Hiroshi Kobayashi, Pavithra Ranganathan, Ayman Mahran, Rakesh Sharma, Anthony J. Thomas, Ashok Agarwal, The Cleveland Clinic Foundation Objective: Sperm quality decreases significantly following freezing and research on improving cryosurvival rates is crucial. We compared the effects of two cryopreservation protocols to determine which method allows better preservation of sperm characteristics.

Oxidative stress and increased levels of apoptosis (Cytochrome C, Caspase 3 and 9) in patients with male-factor infertility.

(CYTOCHROME C, CASPASE 3 AND 9) IN PATIENTS WITH MALEFACTOR INFERTILITY Xia Wang, Rakesh K. Sharma, Pavithra Ranganathan, Anthony J. Thomas, Tommaso Falcone, Ashok Agarwal, The Cleveland Clinic Foundation Objective: Increased production of reactive oxygen species (ROS) results in abnormal seminal quality, DNA damage and the inability of the spermatozoa to fertilize. Mitochondria produces ROS during oxidative phosphorylation, and mitochondrial cytochrome C, a trigger of apoptosis activates caspase 9 and 3 resulting in mitochondrial and subsequently nuclear DNA damage. The objective of the study was to determine the pathway of apoptosis leading to mitochondrial DNA damage. We evaluated, 1) the apoptosis factors in the semen of patients with male-factor infertility, and 2) the association between semen quality and apoptosis in the presence of oxidative stress.

Correlation of nuclear factor kappa B (NFKB) with sperm quality and clinical diagnoses in infertile men

SPERM QUALITY AND CLINICAL DIAGNOSES IN INFERTILE MEN Pavithra Ranganathan, Namita Kattal, Mohamed H Moustafa, Rakesh K Sharma, Anthony J Thomas, Ashok Agarwal, Cleveland Clinic Foundation Objective: NFΚB (p65, p50 and IΚB) plays a major role in regulating apoptosis. Its role in the pathogenesis of male infertility has never been studied. The objectives of our study were to examine 1) the relationship of NFΚB with semen quality (sperm concentration, motility and morphology) and the clinical diagnoses of male infertility, and 2) correlate levels of NFΚB and apoptosis in ejaculated human spermatozoa

Decreased expression of P65, P50 and I kappa B in ejaculated spermatozoa from infertile men

EJACULATED SPERMATOZOA FROM INFERTILE MEN Pavithra Ranganathan, Namita Kattal, Mohamed H Moustafa, Ramadan A Saleh, Rakesh K Sharma, Ashok Agarwal, Cleveland Clinic Foundation Objective: Recent studies indicate high levels of apoptosis in ejaculated spermatozoa from infertile men. Nuclear factor Kappa B (NFΚB) family is an important regulator of apoptosis. It is composed of the sub units p65, p50, and IΚB and is sequestered in the cytoplasm in an inactive form. Recent studies have shown that NFΚB is present in an active form in the nuclei of Sertoli cells in rat testis. However, the activity of NFΚB in ejaculated human spermatozoa has never been investigated. The objective of our study was to examine levels of p65, p50 and IΚBΑ in ejaculated spermatozoa from infertile men as compared to a group of normal sperm donors.

Utilization rate and fertility outcome of cryopreserved sperm from oncological patients: American experience

CRYOPRESERVED SPERM FROM ONCOLOGICAL PATIENTS: AMERICAN EXPERIENCE Pavithra Ranganathan, Ashok Agarwal, Saba Khayal, Namita Kattal, Cleveland Clinic Foundation Objective: The worldwide review of scientific literature reveals a paucity of information regarding the utilization and fertility outcome of cryopreserved semen samples from patients with cancer by assisted reproductive techniques (ART). Our study examined the results of the utilization of cryopreserved semen specimens by oncological patients for reproductive purposes.

Efficacy and compliance of early use vacuum constriction device for erectile dysfunction following radical prostatectomy

CONSTRICTION DEVICE FOR ERECTILE DYSFUNCTION FOLLOWING RADICAL PROSTATECTOMY Rupesh Raina, Ashok Agarwal, Saba Khayal, Pavithra Ranganathan, Namita Kattal, Craig D Zippe, Cleveland Clinic Foundation Objective: To assess the efficacy and compliance of vacuum constriction device (VCD) following radical prostatectomy (RP) and to determiner whether its early use can potentially prevent corporeal fibrosis and expedite recovery of erectile function.

Recovery and survival of sperm is higher with Puresperm density gradient than swim-up in neat and cryopreserved-thawed semen specimens.

PURESPERM DENSITY GRADIENT THAN SWIM-UP IN NEAT AND CRYOPRESERVED-THAWED SEMEN SPECIMENS Pavithra Ranganathan, Ashok Agarwal, The Cleveland Clinic Foundation Objective: Successful assisted reproduction requires sperm processing methods yielding highly motile sperm that can sustain their motility for long periods for fertilization. Our study compared the efficacy of the density gradient method using Puresperm with the traditional swim-up method. The main objectives were to 1) evaluate the motility, longevity, and recovery rates of sperm processed with Puresperm and by the swim-up methods, 2) compare the post-thaw sperm count, motility, recovery rates and longevity between the two methods, and 3) compare sperm characteristics in asthenospermic specimens prepared by the two methods.

Comparison of two sperm counting chambers: microcell and standard count.

MICROCELL AND STANDARD COUNT Hiroshi Kobayashi, Pavithra Ranganathan, The Cleveland Clinic Foundation, Nam-Cheol Park, Pusan National University, James Chae, Ayman Mahran, Ashok Agarwal, The Cleveland Clinic Foundation Objective: Semen analysis remains an essential test for evaluation of male infertility. The main components of semen analysis are sperm count and motility. MicroCell counting chambers are proven devices for accurate measurement of sperm characteristics, while Standard Count was introduced recently. The purpose of this study was to compare the accuracy of MicroCell and Standard Count chambers in measuring sperm count and percent motility.

Assessment of laboratory variability in the measurement of total non-enzymatic antioxidant capacity of semen using an enhanced chemiluminescence assay.

MEASUREMENT OF TOTAL NON-ENZYMATIC ANTIOXIDANT CAPACITY OF SEMEN USING AN ENHANCED CHEMILUMINESCENCE ASSAY Ramadan A. Saleh, Hiroshi Kobayashi, Pavithra Ranganathan, Rakesh K. Sharma, David R. Nelson, Ashok Agarwal, The Cleveland Clinic Foundation Objective: It has been established that non-enzymatic antioxidants in seminal plasma are as important as enzymatic antioxidants in protecting the sperm against oxidative stress-induced damage. Accurate and reliable assessment of total non-enzymatic antioxidant capacity (TAC) of seminal plasma is essential for both research and clinical purposes. The objectives of this study were to assess the inter-donor (different donor samples measured in the same day), intra-assay (running the same sample in duplicate or triplicate), inter-assay (same sample measured on different days by the same observer), and inter-observer (multiple observers on the same day with the same sample) variability using an enhanced chemiluminescence assay.