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Ashok Agarwal

Publications and source records attributed to Ashok Agarwal.

At least 91 records · Page 5Linked to original sources
PubMed2004

Role of free radicals in female reproductive diseases and assisted reproduction.

Infertility is a common problem experienced by many couples. Numerous treatments are available for female infertility. However, in some cases, the treatment is empirical in nature because the aetiology of infertility is not fully understood. Recently, reactive oxygen species (ROS) have been shown to have an important role in the normal functioning of reproductive system and in the pathogenesis of infertility in females. Reactive oxygen species may also play a role in other reproductive organ diseases of women such as endometriosis. Oxidative stress develops when there is an imbalance between the generation of ROS and the scavenging capacity of antioxidants in the reproductive tract. It affects both natural and assisted fertility. Because assisted reproductive techniques are used extensively in the treatment of infertility, it is critical to understand the in-vitro conditions that affect fertilization and embryo development. Treatments that reduce oxidative stress may help infertile women with diseases that are caused by this imbalance. Such strategies include identifying the source of excessive generation of ROS, treating the primary cause, and in-vitro and in-vivo supplementation of antioxidants. Research is in progress to identify the mechanisms that are involved in the aetiology of female reproductive diseases caused by ROS, and to create effective strategies that can counteract oxidative stress.

Antioxidants↗
PubMed2004

Relationship between ROS production, apoptosis and DNA denaturation in spermatozoa from patients examined for infertility.

BACKGROUND: The aim of this study was to examine the role of apoptosis and reactive oxygen species (ROS) in inducing DNA damage in ejaculated spermatozoa. METHODS: We examined ejaculated spermatozoa from 31 patients examined for infertility and 19 healthy donors for apoptosis, production of ROS and DNA damage using annexin V binding, chemiluminescence assay and sperm chromatin structure assay. RESULTS: The percentage of spermatozoa that underwent apoptosis in the whole ejaculate and mature fraction was higher in the patients than in the donors (P<0.001 and P=0.009, respectively). Levels of ROS in the whole ejaculate and immature fraction were higher in the patients than in the donors (P=0.002 and P=0.009). Apoptosis was significantly correlated with ROS within patients in the whole ejaculate [r (95% confidence interval)=0.53 (0.19-0.86)] and in the mature [0.71 (0.39-1.00)] and immature spermatozoa [0.75 (0.45-1.00)]. Only apoptosis and the DNA fragmentation index (DFI) were significantly correlated within patients in the whole ejaculate [0.57 (0.18-0.97)]. CONCLUSIONS: DNA damage may be induced by oxidative assault. Apoptosis may not contribute significantly to the DNA damage.

Antioxidants↗
PubMed2004

[Determination and clinical significance of immature germ cells in human semen].

OBJECTIVE: To discuss the clinical significance of determining immature germ cells (IGC) in human semen. METHODS: Discontinuous Percoll gradients technique was employed to separate different cells and May-Grunwald-Giemsa staining and fluorescein isothiocyanate (FTTC)-Mab-CD45 was adopted to identify IGCs and leukcocytes in semen. The IGCs in 30 semen samples were determined including 10 fertile and 20 infertile cases. RESULTS: IGCs concentrated in gradient fractions with 30% to 45% Percoll and leukocytes concentrated in 50%-55% Percoll fractions. The concentration of IGCs was (0.70 +/- 0.40) x 10(6) ml in the fertile group and (1.28 +/- 0.70) x 10(6)/ml in the infertile group(P < 0.05). There was no statistical correlation between the IGC concentration and the sperm density, vitality and normal morphology(P > 0.05). CONCLUSION: The use of the discontinuous Percoll gradient method can reach the best separation of IGCs in the ejaculate and it is possible to be used as a clinical index to reflect semen quality.

Cell Separation↗
PubMed2004

Relationship between acrosin activity of human spermatozoa and oxidative stress.

AIM: To study the association between seminal oxidative stress and human sperm acrosin activity. METHODS: It is a prospective study consisting of 30 infertile men and 12 fertile normozoospermic volunteers. A full history, clinical examination and scrotal ultrasound were done to exclude other related factors such as smoking and varicocele. Presence of white blood cells (WBCs) in semen samples was evaluated by peroxidase staining. Lipid peroxidation in spermatozoa was induced after incubating with ferrous sulphate (4 mmol/L) and sodium ascorbate (20 mmol/L). Induced peroxidation of spermatozoa was assessed by determining the production of thiobarbituric acid reactive substances (TBARS). Acrosin activity was measured using the gelatinolysis technique. The halo diameters around the sperm heads and the percentages of spermatozoa showing halo formation were evaluated. An acrosin activity index was calculated by multiplying the halo diameter by the halo formation rate. RESULTS: A significant difference was observed in acrosin activity parameters and TBARS levels between samples with WBCs (1 multiply 10(6)/mL of ejaculate) and those without. This difference was also noted between the normozoospermic and the oligoasthenoteratozoospermic semen samples. The TBARS production by spermatozoa had a significant negative correlation with the acrosin activity index (r = -0.89, P 0.001). CONCLUSION: The presence of oxidative stress in an individual with leukocytospermia and/or abnormal semen parameters is associated with impaired sperm function as measured by its acrosin activity.

Acrosin↗
PubMed2003

Detection of testicular cancer in men presenting with infertility.

PURPOSE: Infertility is one of the less common presenting features associated with testicular tumors. We evaluated the histologic and biochemical findings, and pregnancy outcome in patients presenting with infertility who were found to have testicular tumors. METHODS: Seven patients with infertility were found to have testicular cancer over a 15-year period. All patients had a testicular ultrasound evaluation. The indications for the ultrasound were testicular pain in 2 patients, suspicious palpable mass in 4, and to rule out the presence of germ cell neoplasia in a patient with carcinoma in situ detected on a previous biopsy. Physical exam, histological findings, hormonal levels, tumor markers, and pregnancy outcome results were recorded from the patients medical charts. RESULTS: Two men had elevated serum follicle stimulant hormone and luteinizing hormone levels, 1 of them had an abnormally low serum testosterone level. Tumor markers were normal in all patients. In 4 patients the tumor was on the right side and in 3 on the left. The histological diagnoses were seminoma (n = 5), Leydig cell tumor (n = 1), and carcinoma in situ (n = 1). Of the 7 patients, 5 underwent adjuvant radiation therapy. Two patients had sperm cryopreserved. Follow up on fertility status was available in 6 cases. One patient has established a pregnancy and 5 did not achieve a pregnancy after treatment for their cancer. CONCLUSIONS: Most of the men who have testicular cancer and male infertility have a seminona. Therefore, men who present with infertility should be thoroughly investigated to rule out such serious, concomitant diseases along with their infertility.

Adult↗
PubMed2003

Sexual dysfunction in men undergoing infertility evaluation: a cohort observational study.

OBJECTIVE: To study psychosexual problems in men undergoing infertility evaluation. DESIGN: A cohort observational study. SETTING: Male infertility diagnostic center. PATIENT(S): Four hundred twelve men undergoing infertility evaluation between 1999 and 2001. INTERVENTION(S): Baseline and follow-up data on sexual functions. Semen analysis for samples collected by masturbation. A second analysis was requested in 2 weeks upon finding an abnormality of semen parameters. Penile vibratory stimulation to help men who failed to collect semen on their second or subsequent appointments. MAIN OUTCOME MEASURE(S): Sexual functions (erection and orgasm) during the time of infertility evaluation. RESULT(S): Seven of 412 men were excluded from the analysis due to a past history of sexual dysfunction. Of the remaining 405 men, 46 (11%) failed to collect semen by masturbation for a second semen analysis after repeated (2-4 times) attempts at 2- to 3-day intervals. Nine of these men (20%) were able to collect semen using vibratory stimulation. All 46 men experienced problems with erection or orgasm and had severe anxiety during attempts to masturbate and during sexual contact with their partners. CONCLUSION(S): Our study indicates that some men may experience sexual dysfunction of a psychogenic nature in response to the diagnosis of infertility.

Adult↗
PubMed2003

Role of reactive oxygen species in the pathophysiology of human reproduction.

OBJECTIVE: To summarize the role of reactive oxygen species (ROS) in the pathophysiology of human reproduction. DESIGN: Review of literature. SETTING: Fertility research center and obstetrics and gynecology department in a tertiary care facility. RESULT(S): ROS plays an essential role in the pathogenesis of many reproductive processes. In male-factor infertility. oxidative stress attacks the fluidity of the sperm plasma membrane and the integrity of DNA in the sperm nucleus. Reactive oxygen species induced DNA damage may accelerate the process of germ cell apoptosis, leading to the decline in sperm counts associated with male infertility. ROS mediated female fertility disorders share many pathogenic similarities with the ones on the male side. These similarities include a potential role in the pathophysiology of endometriosis and unexplained infertility. High follicular fluid ROS levels are associated with negative IVF outcomes, particularly in smokers. Moreover, oxidative stress may be responsible in hydrosalpingeal fluid mediated embryotoxicity as well as poor in vitro embryonic development. CONCLUSION(S): High levels of ROS are detrimental to the fertility potential both in natural and assisted conception states.

DNA Damage↗
PubMed2003

Negative effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility.

OBJECTIVE: To examine the effects of increased sperm DNA damage in relation to seminal oxidative stress in men with idiopathic and male factor infertility. Prospective study. SETTINGS: Infertility clinic at a tertiary care academic institution. PATIENT(S): Ninety-two infertile men with normal female partners. Sixteen fertile donors served as the control group. INTERVENTION(S): Standard semen analysis and assessment of levels of seminal oxidative stress. Assisted reproductive techniques in 33 of the 92 patients (IUI [n = 19], IVF [n = 10], and intracytoplasmic sperm injection [n = 4]). MAIN OUTCOME MEASURE(S): Sperm DNA damage by sperm chromatin structure assay. Results were expressed as DNA fragmentation index. RESULT(S): Patients were classified as having either idiopathic (n = 23) or male factor infertility (n = 69). Patients with idiopathic and male factor infertility had significantly higher DNA fragmentation index and oxidative stress compared with the case of fertile donors. A clinical pregnancy was achieved in 9 (27%) of 33 patients with assisted reproductive techniques. Significantly higher DNA fragmentation index and oxidative stress were found in men who failed to initiate a pregnancy after assisted reproductive techniques (n = 24), compared with the cases of those who succeeded and of the fertile donors. DNA fragmentation index was correlated positively with oxidative stress (r = 0.27), and negatively with fertilization (r = -0.70) and embryo quality (r = -0.70). CONCLUSION(S): Sperm DNA damage is significantly increased in men with idiopathic and male factor infertility and in men who failed to initiate a pregnancy after assisted reproductive techniques. Such an increase may be related to high levels of seminal oxidative stress.

Adult↗
PubMed2003

Oxidative stress is associated with increased apoptosis leading to spermatozoa DNA damage in patients with male factor infertility.

OBJECTIVE: To evaluate ejaculated spermatozoa from patients with male factor infertility for the role of cytochrome c and caspases 9 and 3 (the proteins known to mediate apoptosis) and to examine association between semen quality and apoptosis in the presence of oxidative stress. DESIGN: Prospective study. SETTING: Male infertility clinic at a tertiary health care center. PATIENT(S): Semen specimens from 35 patients with idiopathic infertility and 8 normal healthy donors. MAIN OUTCOME MEASURE(S): Levels of ROS, cytochrome c, and caspases 9 and 3 and semen variables. RESULT(S): Compared with normal donors, infertile patients had significantly higher levels (expressed as median [25th and 75th percentiles]) of ROS (4.15 x 10(6) counted photons per minute [cpm] [0.26, 40.16 x 10(6) cpm] vs. 0.06 x 10(6) cpm [0.02, 0.29 x 10(6) cpm]; P<.01), cytochrome c (2.78 [2.21, 43.65 vs. 1.5 [1.25, 2.2]; P<.01)], caspase 9 (2.52 [0.9, 4.28] vs. 6 [4.85, 7.63]; P<.006), and caspase 3 (0.56 [0.32, 1.02] vs. 1.69 [1.66, 2.67]; P<.01). Semen variables (motility, concentration, and morphology) were negatively correlated with caspase 9 and 3 (P<.05). Reactive oxygen species was positively correlated with cytochrome c (r=.43; P<.03), caspase 9 (r=.56; P<.001), and caspase 3 (r=.65;P<.01). CONCLUSION(S): Infertile men have decreased sperm variables induced by higher ROS levels in semen. A positive relationship exists between increased sperm damage by ROS and higher levels of cytochrome c and caspases 9 and 3, which indicate positive apoptosis in patients with male factor infertility.

Apoptosis↗
PubMed2003

Alterations in mitochondria membrane potential and oxidative stress in infertile men: a prospective observational study.

OBJECTIVE: To evaluate the mitochondrial membrane potential (MMP) of spermatozoa and its correlation with semen parameters and production of reactive oxygen species (ROS) in infertile men and healthy donors. DESIGN: Controlled prospective study. SETTING: Male infertility clinic, Glickman Urological Institute, The Cleveland Clinic Foundation, Cleveland, Ohio. PATIENT(S): Nineteen infertile men and 7 healthy volunteers. INTERVENTION(S): Standard semen analysis, assessment of MMP and ROS production in spermatozoa. The MMP was assessed by flow cytometry using the probe carbocyanine DiOC(6)(3) and ROS was measured with chemiluminescence assay using luminol. MAIN OUTCOME MEASURE(S): The results of MMP are reported as the median interquartile range (IQR) number of cells counted in different areas of fluorescence. Results of ROS measurement are expressed as x10(6) counted photons per minute per 20 million sperm (cpm). RESULT(S): The patients with abnormal semen parameters had a significantly lower MMP [1337.7 (1066.38, 1879.2)], and higher ROS [1.12 (0.26, 3.86)] than the donors [MMP: 2482.9 (2162.5, 3520.6)] and [ROS: 0.10 (0.01, 0.14)]. The MMP was positively correlated with sperm concentration (r = 0.62) and negatively correlated with the ROS produced (r = -0.45). CONCLUSION(S): Measuring MMP in spermatozoa provides useful information about a man's fertility potential. Increased ROS production by spermatozoa is associated with a decreased MMP.

Carbocyanines↗
PubMed2003

Evaluation of nuclear DNA damage in spermatozoa from infertile men with varicocele.

OBJECTIVE: To examine levels of sperm DNA damage and oxidative stress (OS) in infertile men with varicocele. DESIGN: Prospective controlled study. SETTING: Male infertility clinic, Glickman Urological Institute, Cleveland Clinic Foundation, Cleveland, Ohio. PATIENT(S): Thirty-one infertility patients and 16 fertile controls. INTERVENTION(S): Sperm DNA fragmentation index (DFI), levels of seminal reactive oxygen species (ROS), and total antioxidant capacity (TAC) were assessed using the sperm chromatin structure assay, chemiluminescence assay, and enhanced chemiluminescence assay, respectively. ROS-TAC score was calculated as a measure of OS. MAIN OUTCOME MEASURE(S): Median (interquartile range) DFI and ROS-TAC scores. RESULT(S): Sixteen of the 31 patients had left varicocele [grade I (n = 3), grade II (n = 10), and grade III(n = 3)], and the remaining 15 had normal genital examination. Patients with varicoceles had significantly higher percent DFI than controls (25%, range: 20%-35%; vs. 15%, range: 10%-22%). Patients with varicoceles had significantly lower ROS-TAC scores (21, range: 9.5-31) than the infertile patients with normal genital examination (34, range: 28-42) or the controls (40.3, range: 38-44). CONCLUSION(S): Infertile men with varicoceles showed significantly increased spermatozoal DNA damage that appears to be related to high levels of OS in semen.

Cell Nucleus↗
PubMed2003

Long-term effect of sildenafil citrate on erectile dysfunction after radical prostatectomy: 3-year follow-up.

OBJECTIVES: To evaluate the long-term effect and safety of sildenafil citrate for the treatment of erectile dysfunction after radical prostatectomy (RP). METHODS: The study consisted of 91 patients with erectile dysfunction from our institution who received oral sildenafil citrate after RP. We surveyed these patients using a self-administered questionnaire during the first year of sildenafil citrate use to determine treatment satisfaction, patient compliance, and safety. Those who had responded positively to the drug were surveyed again 3 years later (n = 48). Sildenafil citrate was prescribed at a dose of 50 mg and increased to 100 mg if needed. Data were collected from a self-administered questionnaire using the abridged five-item version of the International Index of Erectile Function questionnaire, referred to as the Sexual Health Inventory of Men, and the Erectile Dysfunction Inventory of Treatment Satisfaction. The patients were stratified according to the type of nerve-sparing (NS) RP procedure they underwent: bilateral NS, unilateral NS, and non-NS. RESULTS: At 3 years, 31 (71%) of the 43 patients who had returned the second surveys were still responding to sildenafil. Of these 31 respondents, 10 (31%) had augmented their dose from 50 to 100 mg. The dropout rate was 27%; 6 of 12 had discontinued because of the return of natural erections, 5 because of a loss of efficacy, and 1 because his spouse had died. No differences were found in the 1-year and 3-year five-item International Index of Erectile Function (Sexual Health Inventory of Men) and Erectile Dysfunction Inventory of Treatment Satisfaction scores between the NS groups. The most common side effects at 3 years were headache (12%), flushing (10%), and blue or blurred vision (2%). No patient discontinued the drug at 3 years because of side effects. CONCLUSIONS: The results of this study indicate that the vast majority of patients with erectile dysfunction after RP who initially respond to sildenafil continue to do so at 3 years and are satisfied and compliant with the treatment regimen.

3',5'-Cyclic-GMP Phosphodiesterases↗
PubMed2003

Long-term potency after iodine-125 radiotherapy for prostate cancer and role of sildenafil citrate.

OBJECTIVES: To assess the long-term sexual potency and attrition in sexual function after iodine-125 ((125)I) seed radiotherapy and the effect of sildenafil on radiation-induced erectile dysfunction (ED). METHODS: This prospective study consisted of 86 sexually active patients (mean age 63.5 +/- 7.7 years) who underwent (125)I seed implantation from 1997 to 1999 to treat low-volume prostate cancer (prostate-specific antigen less than 10 ng/mL, Gleason score 6 or less, stage T1-T2). All patients were followed up every 6 to 8 months for 4 years. Patients prescribed sildenafil citrate for ED completed the abridged five-item version of the International Index of Erectile Function (IIEF) and the Erectile Dysfunction Inventory of Treatment Satisfaction (EDITS) questionnaires. RESULTS: The median follow-up was 49.7 months (range 36 to 66). Of 86 patients, 43 (50%) did not initiate drug therapy; and only 36 (83.7%) of the 43 were interviewed at 4 years. Twenty-three (63.8%) of the 36 patients had erections sufficient for vaginal penetration, with a total mean +/- SD IIEF-5 score of 15.76 +/- 1.13. The other 50% (43 of 86) initiated sildenafil citrate for treatment of ED after seed implantation, with a minimal follow-up of 6 months. At 4 years, 32 (74%) of the 43 were responding positively to sildenafil citrate, with a total IIEF-5 score of 18.3 +/- 1.2. The mean EDITS +/- SD score was 76.5 +/- 3.2, and the spousal satisfaction rate was 72% (31 of 43). The dropout rate was 37% (16 of 43); 10 (63%) of the 16 discontinued because of a lack of efficacy, 3 (19%) because of a return of natural erections sufficient for vaginal penetration, and 3 (19%) discontinued because of side effects (headaches). CONCLUSIONS: ED is a major long-term issue after (125)I seed radiotherapy, with a long-term potency rate of 29%. Sildenafil citrate improves erections in most patients after (125)I seed implantation.

Adenocarcinoma↗
PubMed2003

Lower sperm aneuploidy frequency is associated with high pregnancy rates in ICSI programmes.

BACKGROUND: A large proportion of patients undergoing ICSI have been shown to have an increased sperm aneuploidy rate. This study was undertaken to evaluate the impact of sperm aneuploidy on ICSI outcome. METHODS: To accomplish this, 48 consecutive unselected male patients (median age 34 years) had their sperm aneuploidy rate evaluated in the same swim-up preparation used for ICSI. Chromosomes 8, 12, 18, X and Y were evaluated by fluorescence in-situ hybridization. Patients were divided into two groups (A and B) based on the sperm aneuploidy frequency in their sperm. Group A had values below and group B above the upper limit of normal [1.55%, determined in 14 healthy men (median age 25 years) with normal semen parameters by WHO 1999 criteria (control group)]. RESULTS: Group A consisted of 12 patients (25%) whose sperm aneuploidy rates fell below the cut-off value of the control group (median 1.25%; range 0.85-1.52). Group B consisted of the remaining 36 patients (75%), who had an elevated sperm aneuploidy rate (median 3.25%; range 1.64-23.60). Fertilization (93 versus 85%) and cleavage (100 versus 98%) rates were similar for both groups. Group A had significantly higher clinical pregnancy (75 versus 34%; P < 0.001) and implantation (34 versus 13%; P < 0.001) rates compared with group B. In addition, group A had a lower overall miscarriage rate (11.1 versus 38.9%). Other factors that affect pregnancy and implantation, such as patient age and conventional semen parameters, were similar for both groups. CONCLUSION: This study showed that chromosomally abnormal sperm have a negative impact on ICSI outcome.

Abortion, Spontaneous↗
PubMed2002

Increased DNA damage in sperm from leukocytospermic semen samples as determined by the sperm chromatin structure assay.

OBJECTIVE: To determine DNA damage as measured by the sperm chromatin structure assay (SCSA) in subsets of human spermatozoa at different stages of maturation in patients who are undergoing infertility evaluation. DESIGN: Prospective study. SETTING: Andrology laboratory at a tertiary care hospital. PATIENT(S): Fifty-six patients undergoing infertility evaluation. Patients with normal semen parameters (n = 17), abnormal semen parameters (n = 29), leukocytospermia (n = 10), and a group of healthy fertile men (n = 18) were included in the study. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The shift of green (native DNA) to red (denatured, single-stranded DNA) fluorescence was measured and quantified using the expression alpha(t) (red fluorescence/[red + green fluorescence] per cell). Sperm DNA damage was examined in subsets of spermatozoa isolated by a three-step density gradient. The DNA damage was correlated with classic semen characteristics. RESULT(S): Leukocyte concentration in semen was directly correlated with chromatin alterations in immature and mature sperm. Leukocyte concentration in semen was also directly correlated with immature germ cell concentration and the percentage of abnormal forms in semen. CONCLUSION(S): The increase in chromatin alterations and DNA damage in sperm, as defined by the sperm chromatin structure assay from leukocytospermic samples may be related to alterations in the regulation of spermatogenesis.

Centrifugation, Density Gradient↗
PubMed2002

Increased sperm nuclear DNA damage in normozoospermic infertile men: a prospective study.

OBJECTIVE: To evaluate levels of sperm nuclear DNA damage in infertile men with normal and abnormal standard semen parameters. DESIGN: Prospective study. SETTING: Male infertility clinic. PATIENT(S): Ninety-two men seeking infertility treatment and 16 fertile volunteers. INTERVENTION(S): Standard semen analysis was performed according to the World Health Organization guidelines. MAIN OUTCOME MEASURE(S): Sperm DNA damage was assessed by sperm chromatin structure assay and the results expressed as %DFI. RESULT(S): Of the 92 patients, 21 (23%) had normal standard sperm parameters (concentration, motility, and normal sperm forms), while 71 (77%) had an abnormality in one or more of these parameters. The %DFI [median (25th and 75th percentiles)] in infertile men with normal sperm parameters [23 (15, 32)] was significantly higher than fertile donors [15 (11, 20)] (P=.02), but not significantly different from infertile men with abnormal sperm parameters [28 (18, 41)] (P=.27). CONCLUSION(S): The results of this study indicate that a significant increase in SCSA-defined DNA damage can be found in sperm from infertile men with normal standard sperm parameters. Therefore, sperm DNA damage analysis may reveal a hidden abnormality of sperm DNA in infertile men classified as idiopathic based on apparently normal standard sperm parameters.

Cell Nucleus↗