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Purpose: We evaluated the morphological, immunological and functional response to small intestinal submucosa grafting of the tunica albuginea to determine its potential as a grafting material for penile surgery., Materials and Methods: Male New Zealand White rabbits underwent a sham procedure (6) or tunical excision and grafting with small intestinal submucosa (6). The erectile response to the intracavernous vasoactive agents sodium nitroprusside plus a papaverine, phentolamine and prostaglandin E1 combination (Sigma Chemical Co., St. Louis, Missouri) was evaluated 45-day postoperatively. The area under the graft was evaluated for stromal collagen and smooth muscle content by Masson's trichrome stain. Protein expression of smooth muscle specific alpha-actin and the inflammatory markers inducible nitric oxide synthase (NOS) and transforming growth factor-beta1 (TGF-beta1) was evaluated by immunohistochemical methods. Total RNA was extracted from the corpora cavernosum underlying the small intestinal submucosa graft and reverse transcriptase-polymerase chain reaction (RT-PCR) was done using an Access system (Promega, Madison, Wisconsin) with gene specific primers for inducible NOS, TGF-beta1 and vascular endothelial growth factor (VEGF)., Results: Grafting of the tunica albuginea with small intestinal submucosa had no significant effect on the magnitude or duration of the erectile response to intracavernous vasoactive agents. Histological examination demonstrated no inflammatory changes in the tunica albuginea or corporeal tissue underlying the area of the small intestinal submucosa graft and there was no appreciable alteration in smooth muscle or collagen content. The 2 groups showed intense positive immunostaining to alpha-actin. Weak expression of TGF-beta1 predominantly associated with smooth muscle fibers was identified in the 2 groups of rabbits by immunostaining and RT-PCR. No significant inducible NOS was detected by immunostaining or RT-PCR in either group. Strong VEGF expression was observed in grafted rabbits. The most noticeable (3-fold) increase in expression was detected in splice variant 165., Conclusions: Small intestinal submucosa grafting of the tunica albuginea preserves the duration and magnitude of the erectile response to vasoactive agents. This type of tunical grafting does not stimulate a significant inflammatory response, or cause corporeal fibrosis or loss of cavernous smooth muscle content. Stimulating VEGF may facilitate wound healing and the maintenance of normal erectile function.

The objective of the present study was to evaluate age-related changes in the protein and gene expression of modulators of erectile function (nitric oxide [NO] and endothelin-1 [ET-1]) and growth factors such as transforming growth factor (TGF-beta1) and vascular endothelial growth factor (VEGF) in the penile tissue of Brown-Norway (BN) rats. Young and old BN male rats were euthanized, and the penile tissue was processed for immunohistochemical and molecular analyses. Total RNA was extracted, and an Access reverse transcription-polymerase chain reaction (RT-PCR) system was used for messenger RNA (mRNA) expression analysis. Immunohistochemical studies showed a decreased expression of endothelial nitric oxide synthase (eNOS) protein and an increased staining for ET-1. Quantitative analysis of PCR products revealed decreased levels of VEGF mRNA expression in the old population of rats. The most significant decrease was detected between bands corresponding to splice forms 164 (21%) and 120 (18%). The observed alterations in the gene expression of growth factors such as VEGF may contribute to the abnormal age-related morphological and physiological alterations in the erectile tissue.

Male sexual function requires an intricate interplay between the man and his environment. Cognitive integration and physiological response to sexual stimulation is dependent on complex neurologic functions that may be impaired by central or peripheral neurologic disorders. This article reviews the normal neuroanatomy of sexual functioning in men, and the epidemiology, pathophysiology and management of sexual dysfunction in spinal cord injury, cerebrovascular accident, multiple sclerosis and Parkinson's disease.

Spinal cord injury (SCI) commonly affects males in their reproductive years. After spinal cord injury, most men experience fertility related problems including erectile and ejaculatory dysfunction, impaired spermatogenesis, abnormal sperm viability, motility, and morphology, genitourinary infection and endocrine abnormalities. In this article we will review the pathophysiology, evaluation and management of infertility in spinal cord injury. The impact of spinal cord injury on seminal plasma and the contribution of seminal oxidative stress to the poor sperm quality of men with spinal cord injury will be presented. Success with sperm retrieval techniques and assisted reproductive technology in SCI men with spinal cord injury will be discussed.

Objectives: To evaluate the direct in vitro cytotoxicity of vasoactive agents (papaverine, phentolamine, and prostaglandin E(1) [PGE(1)]) to human penile cavernosal smooth muscle cells. Intracavernous pharmacotherapy with vasoactive agents for male erectile dysfunction has been associated with long-term complications such as a reduction in penile smooth muscle content and fibrosis., Methods: Human penile cavernosal tissue explants (1 to 2 mm(3) size) were obtained with proper institutional review board approval from patients undergoing penile prosthesis implantation. Primary culture was initiated in Dulbecco's modified Eagles medium containing 10% fetal bovine serum, and monolayer cavernosal cells were grown in 48-well tissue culture dishes. At 60% to 80% confluence, cells were labeled overnight with (51)Na(2)CrO(4) (1.5 microCi) and then incubated with therapeutic concentrations of papaverine (1.5 to 30 mg/mL), phentolamine (0.5 mg/mL), and PGE(1) (5 microg/mL) alone, as well as in combination, for 30 minutes at 37 degrees C. At the end of incubation, an aliquot of supernatant was collected in scintillation vials. The release of cell-free chromium in supernatants was determined in a liquid scintillation counter, and results were expressed as the percentage of cytotoxicity., Results: Papaverine induced a significant dose-dependent increase in chromium release from the cavernosal cells. At therapeutic concentrations, papaverine (30 mg/mL) produced up to 60% cytotoxicity; PGE(1) (5 microg/mL) resulted in 40% toxicity. The combination of papaverine with either PGE(1) or phentolamine had a cumulative toxic effect, and maximal toxicity (70%) was observed with the triple combination., Conclusions: Papaverine-induced cytotoxicity to cavernosal smooth muscle cells may contribute to the fibrosis and loss of smooth muscle content associated with the intracavernous pharmacotherapy. Quantitative evaluation of in vitro cytotoxicity in human cavernosal smooth muscle cell culture may be important in the development of new intracavernosal vasoactive agents.

Pro-inflammatory cytokines are elevated in the semen of patients with genitourinary inflammation (GUI). Whether this increase in cytokines in GUI patients plays any critical role in male factor infertility is not clear. The present study investigated the in vitro effects of two important pro-inflammatory cytokines, lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma), on sperm motility, viability, membrane integrity and motion parameters. Washed spermatozoa from healthy donors were incubated with LPS (0.1 mg/mL) or IFN-gamma (0.1 mg/mL) alone or in combination. Sperm motility, viability, membrane integrity and computer-assisted motion were evaluated at various time intervals (0, 30, 60 and 180 min) after treatment. Sperm membrane integrity was analysed using the hypo-osmotic swelling test (HOST). LPS and IFN-gamma individually did not alter sperm viability or motility, but their combination showed a significant time-dependent decrease (p < 0.05) in sperm motility, viability and membrane integrity. Sperm motion parameters (straight-line velocity, curvilinear velocity, mean linearity, or amplitude of lateral head displacement) were not affected by LPS or IFN-gamma at the concentrations used in this study. These data suggest that the combination of LPS and IFN-gamma is detrimental to human spermatozoa and may contribute to male factor infertility in patients with chronic GUI.

Background: In men with spinal cord injury (SCI), fertility is impaired because of a combination of ejaculatory dysfunction and poor semen quality. We hypothesized that ultrastructural and metabolic abnormalities of sperm could be an underlying factor in infertility in these men., Methods: To investigate mechanisms contributing to the abnormal sperm parameters, we analyzed seminal constituents and sperm ultrastructure in ejaculates from 7 men with SCI and compared them with 5 control subjects. Sperm adenosine triphosphate (ATP) content was measured using a firefly luciferase bioluminescent assay kit, and ultrastructure was evaluated by electron microscopy. Seminal oxidative stress (8-iso-prostaglandin F-2alpha [8-iso-PGF2alpha]) and transforming growth factor-beta1 (TGF-beta1) levels were measured by specific enzyme-linked immunoabsorbent assay kits., Results: Semen samples from men with SCI showed azoospermia (3) and severe asthenospermia (2; motility 0-5%). A majority (65%) of sperm from asthenospermic samples showed degenerative changes and significant axonemal defects. Incubation of normal sperm with SCI seminal plasma induced a concentration-dependent decrease in sperm motility (43%) accompanied by a significant drop in intracellular ATP content (33%). Semen samples from men with SCI exhibited levels of 8-iso-PGF2alpha 3.5-fold higher than those from controls and levels of TGF-beta1 that were 10% higher than those from controls., Conclusion: Our results suggest that seminal constituents of men with SCI are detrimental to sperm movement and that ultrastructural degenerative changes may contribute to the impaired sperm motility and viability seen in these patients. These preliminary results must be confirmed in larger patient populations and longitudinal studies.

Nitric oxide (NO) is a product of nitric oxide synthase (NOS) activity and is recognized as the main mediator of penile erection by induction of cavernosal smooth muscle relaxation. Although excessive NO can be generated via inducible NOS activation under certain inflammatory and noninflammatory conditions, for example, in response to TGF-beta and gamma-IFN (the proinflammatory cytokines), the effect of excessive NO produced as reactive nitrogen radical (NO.-) in the corpora cavernosa is not known. The present study was designed to evaluate whether the effect of NO.- on human cavernosal cells in primary culture is via oxidative stress. Cell growth was monitored by DNA synthesis, and mitochondrial function was evaluated by adenosine triphosphate (ATP) production. Primary culture was initiated with explants from human corpora cavernosa, and the monolayer cavernosal cells (passage 2-3) were plated on 12-well tissue culture plates. At 70%-80% confluency, the cells were incubated with varying concentrations of sodium nitroprusside (SNP) for 16 hours. The cell growth (DNA synthesis) was monitored by measuring [3H] thymidine incorporation, ATP levels (nanomoles per 10(4) cells) were measured by chemiluminescence assay using a luminometer, the total oxidative stress was monitored by measuring the levels of 8-iso PGF2alpha (picograms per milliliter) by using an enzyme-linked immunosorbent assay kit, and NO production was monitored by accumulation of nitrite levels (micrometer per 10(4) cells). Human cavernosal smooth muscle cells (HCSMC) exposed to SNP (0 to 0.8 mM) exhibited a dose-dependent (two- to fivefold) decrease in DNA and ATP synthesis, accompanied by a two- to threefold increase in the levels of 8-iso PGF2alpha and about an eightfold increase in nitrite accumulation. These findings suggest that the NO released by SNP (>0.8 mM) exhibited a significant cytotoxicity to HCSMC, mediated by increased oxidative stress to these cells.

Purpose: Phosphodiesterases (PDEs) are an important component of the signal transduction pathway during the erectile response. To determine the PDE isoforms in the corpora cavernosa in the cat and to establish the functional presence of PDE 4 in human cavernosal tissue, the erectile response to intracavernosal phosphodiesterase (PDE) inhibitors alone and the combination of PDE inhibitors and prostaglandin E1 (PGE1) was evaluated in the anesthetized cat. The in vitro formation of cAMP and cGMP in human cavernosal smooth muscle cells (HCSMCs) treated with PGE1 and rolipram in primary culture was also measured., Materials and Methods: In pentobarbital-anesthetized cats, increases in intracavernosal pressure, penile length, and duration of erectile response were determined after intracavernosal injections of (i) the type 3 cAMP-specific, cGMP-inhibitable PDE inhibitor, milrinone, (ii) the type 4 cAMP-specific PDE inhibitor, rolipram, (iii) the type 5 cGMP-specific PDE inhibitor, zaprinast, and (iv) the combination of rolipram and PGE1. Systemic arterial pressure was concurrently assessed in these experiments. All responses to PDE inhibitors were compared with a control triple-drug combination comprised of papaverine (1.65 mg.), PGE1 (0.5 microg.), and phentolamine (25 microg.). HCSMCs were incubated with PGE1 (3 microM) and rolipram (10 microM) individually or in combination up to 2 hours at 37C. The intracellular cAMP and cGMP was extracted by cold absolute ethanol and measured (pmol./10(6) cells) by a commercially available EIA kit., Results: Milrinone (3 to 100 microg.), rolipram (3 to 100 microg.), and zaprinast (3 to 100 microg.) induced dose-dependent increases in intracavernosal pressure and penile length (p <0.05) when administered intracavernosally. The maximum increase in cavernosal pressure in response to zaprinast was associated with no significant change in systemic arterial pressure. When rolipram was combined with PGE1 (0.1 microg.), the increases in intracavernosal pressure and the duration of erectile response were significantly higher (p <0.05) and longer (p <0.05) than those observed when rolipram alone was injected intracavernosally. PGE1 (3 microM) and rolipram (10 microM) produced significant increases (p <0.05) in the accumulation of intracellular cAMP levels in HCSMCs in primary culture above those of the baseline values while intracellular levels of cGMP did not change., Conclusions: PDE inhibitors administered intracavernosally caused dose-dependent increases in cavernosal pressure in the cat. When a specific cAMP PDE inhibitor was combined with PGE1, the erectile response was enhanced and intracellular levels of cAMP were increased in HCSMCs in primary culture. These data suggest further exploration of the combination of various PDE inhibitors and PGE1 in the pharmacologic treatment of erectile dysfunction and provide functional evidence for the presence of PDE 4 isoenzyme in human penile cavernosal cells.

Objective: To review the pathophysiology, evaluation, and management of erectile dysfunction and infertility in spinal cord injury (SCI)., Study Selections: Studies that covered various treatment options and their contraindications, complications, or side effects, including sildenafil (Viagra), intracavernosal injection therapy, topical medications and a urethral delivery system, a vacuum erection device, and penile prostheses. Other studies covered the effects of SCI on reproduction: spermatogenesis and testicular function, and seminal constituents. In addition, assisted reproductive techniques were compared: external vibratory stimulation, electroejaculation, testicular sperm aspiration, and intracytoplasmic sperm injection., Conclusion: Goal-directed therapy is the mainstay of treatment of erectile dysfunction in men with or without SCI. The choice of therapy is often defined more by the needs of the patient's sexual relationship than by his erectile dysfunction. The majority of men with SCI are infertile because of a combination of ejaculatory dysfunction, impaired spermatogenesis, and poor semen quality. Although many technological advances have evolved to overcome ejaculatory dysfunction, the sperm density, motility, and function remain poor. Until these parameters are improved, men with SCI will have to pursue more financially and emotionally taxing procedures. Further studies to elucidate the cellular and molecular mechanisms of diminished sperm quality are needed. Addressing the issues of erectile dysfunction and male infertility may help to preserve the relationship between the patient and his partner.

Purpose: Nitric oxide (NO) synthesized by nitric oxide synthase (NOS) is recognized as the central mediator of penile erection. This process appears to be mediated mainly by neuronal NOS (nNOS), which is localized to the nonadrenergic, noncholinergic innervation of the penis. However, the role of non-neuronal penile constituents (specifically the cavernosal smooth muscle), as well as other NOS isoforms in NO production in the human penis is not well understood. The present study evaluates the expression of non-neuronal (inducible and endothelial) isoforms of NOS in human penile cavernosal smooth muscle cells in culture., Materials and Methods: Primary culture was initiated with explants of human corpora cavernosa. For gene expression studies, total RNA was extracted from cavernosal cells and subjected to reverse transcriptase polymerase chain reaction (RT-PCR). For NADPH-diaphorase histochemistry, the cells were incubated with 1 mM beta-NADPH and 0.5 mM nitrobluetetrazolium at 37C for 3 hours. For indirect immunofluorescence and electron microscopy, cells were incubated overnight at 4C with specific primary (eNOS; calmodulin) and secondary antibodies. A conventional avidin biotin complex technique was used for electron microscopy., Results: The mRNA expression studies revealed that these cells express both endothelial (eNOS) and inducible (iNOS) forms. Localization studies showed positive signals for NADPH-diaphorase, eNOS, and calmodulin. The electron microscopic evaluation confirmed the localization of eNOS to the cytoplasm and small vesicles in the cells., Conclusions: These findings support the hypothesis that human cavernosal smooth muscle cells express both endothelial and inducible forms of NOS, which may significantly contribute to NO production in the penile architecture during the erectile process.

The purpose of this study was to investigate the in vivo effects of intracavernosal injections of adrenomedullin (ADM), calcitonin gene-related peptide (CGRP), nociceptin, vasoactive intestinal polypeptide (VIP), sodium nitroprusside (SNP), and prostaglandin E1 (PGE1) on penile erection in castrated and intact (control) anesthetized cats. Erectile responses to ADM, CGRP, nociceptin, VIP, SNP, and PGE1 were compared with responses to a standard triple-drug combination (1.65 mg of papaverine, 25 microg of phentolamine, and 0.5 microg of PGE1) in both castrated and control cats. In control animals, ADM, CGRP, nociceptin, VIP, SNP, and PGE1 induced penile erections similar to those elicited by the triple-drug combination. However, in castrated animals, there was a significant decrease in erectile response; the response to intracavernosal injection of the standard triple-drug combination in castrated cats was 28% of that of the control group of animals. Serum testosterone levels demonstrated a significant (P < 0.0001) positive correlation (r = 0.52) with intracavernosal pressure in response to the standard combination. A marked reduction in serum testosterone levels was observed in castrated cats when measured by radioimmunoassay (0.34 +/- 0.1 ng/dl in castrated cats, compared with 31.15 +/- 6 ng/dl in control cats). These data suggest that the presence of testosterone is a necessary prerequisite to sustain a pharmacologically induced penile erection in the cat.

Human serum albumin (HSA) is being considered as an alternate media for sperm enrichment in assisted reproductive technology (ART) because of recent concern with the use of Percoll. In this study, we compared HSA and Percoll for 1) sperm recovery, 2) reactive oxygen species scavenging potential, and 3) effects on total oxidative stress to spermatozoa. The spermatozoa-enriched fractions obtained from Percoll (80%:40%) and HSA (12%) were monitored for sperm motility, viability, hypoosmotic swelling test (HOST), and adenosine triphosphate (ATP) levels. The effect of superoxide anions (O2.-) on donor human spermatozoa was observed in the presence of either HSA or Percoll media. A combination of luminol and the Cypridina luciferin analog 2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo(1,2-alpha)pyraz in-3-one hydrochloride was used as a highly sensitive chemiluminescence probe in our hypoxanthine and xanthine oxidase-based assay for O2.-. Sperm membrane total oxidative stress was determined by measuring levels of the prostanoid 8-iso-Prostaglandin F2alpha (8-iso-PGF2alpha). Significant differences in sperm parameters between the Percoll-enriched spermatozoa (motility 60%+/-4%, viability 56%+/-6%, and HOST 73%+/-7%) and those enriched with HSA (motility 84%+/-5%, viability 85%+/-4%, and HOST 84%+/-3%; P < 0.01) were observed. Adenosine triphosphate levels were significantly higher, by almost 50%, in samples processed with HSA than with Percoll (P=0.03). The dismutation rate of O2.- in HSA (slope -6.8) was significantly lower than in Percoll (slope -87.0; P < 0.01). Sperm motility and ATP levels decreased at a slower rate after treatment with O2.- in the presence of HSA when compared to Percoll; moreover, spermatozoa in HSA regained partial motility after 2 hours, whereas spermatozoa in Percoll were immobilized. No significant differences in 8-iso-PGF2alpha levels in spermatozoa enriched by either HSA or Percoll were observed. We conclude that the HSA sperm enrichment procedure improves the recovery of higher quality spermatozoa compared to Percoll and, because of its antioxidant properties, may be useful in processing high leukospermia semen samples for ART purposes.

Male genital tract infections and non-specific inflammatory conditions may be associated with unexplained infertility. Previous studies have shown the presence of cytokines such as tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) in the semen of infertile men. However, the mechanism of effect of these cytokines on human sperm function is still controversial. The present study was undertaken to investigate the in-vitro effects of TNF-alpha and IFN-gamma on human sperm motion, viability and the hypoosmotic swelling test (HOST). Washed spermatozoa from normal volunteers (n = 9) were incubated in the presence/absence of TNF-alpha (1 microgram/mL) plus IFN-gamma (0.1 microgram/mL). Sperm motility, viability, HOST, and video sequences were recorded at different time intervals (0, 30, 60 and 180 min). Sperm motion parameters were analysed using computer-assisted semen analysis. There was a time-dependent negative effect of TNF-alpha plus IFN-gamma on sperm motility, viability, HOST, and lateral-head displacement (ALH). The maximum decrease was observed between 60 and 180 min for sperm motility (50.8 +/- 5.6%), viability (52.8 +/- 4.0%), HOST (38 +/- 2%) and ALH (4.7 +/- 0.1 microns) compared to control samples (62.2 +/- 2.8, 62.4 +/- 2.9, 58 +/- 4, and 5.3 +/- 0.4, respectively; All p < 0.05). There was no significant effect on sperm straight-line velocity and mean linearity when compared to control. These data suggest that the common inflammatory cytokines TNF-alpha plus IFN-gamma have only partial detrimental effects on sperm motility, viability, membrane integrity and lateral head displacement, which may contribute to the poor fertilizing potential of human spermatozoa during inflammatory conditions.

The Gunn rat is an excellent model of Crigler-Najjar syndrome, type 1. In previous studies we demonstrated that heterotopic 15-20-cm jejunal transplants from Wistar rats lowered serum bilirubin levels by 40%, and the reduction was transient (6 weeks). In contrast, orthotopic transplants decreased bilirubin levels by 60% and the effect persisted throughout the 8-week study. This study was initiated to identify the luminal substance(s) which are responsible for the persistent bilirubin-lowering effect of jejunal transplants. Thirty-one Wistar to Gunn 15-20-cm jejunal transplants were randomized to receive daily Thiry-Vella graft irrigation with 5 ml of normal saline (n = 8); bile salts (cholate + deoxycholate, 40 mg/ml, n = 5; fats (Microlipid, 20 mg/ml, n = 5); proteins (Casec caseinate, 40 mg/ml, n = 5); and sugars (Moducal + Polycose, 40 mg/ml, n = 8). Bilirubin levels were measured spectrophotometrically at weekly intervals. At 4 and 8 weeks, enzyme-induced bilirubin conjugation activity was measured using added known amounts of added bilirubin. Irrigation of the transplants with saline, protein, and sugar resulted in moderate (40%) lowering of serum total and indirect bilirubin levels. Fat was significantly more effective, lowering mean total bilirubin levels from 9.6 +/- 0.4 to 1.6 +/- 0.2 mg/dl at 6 weeks. After this time, bilirubin levels increased slightly. Bile salts were slightly less effective, lowering bilirubin levels at 6 weeks by only 75%. However, this effect persisted and at 8 weeks levels averaged 2.4 +/- 0.2 mg/dl. Conjugating enzyme activity in the transplants increased from 1.4 +/- 0.3 to 2.5 +/- 0.5 mg bilirubin conjugated/mg tissue/hr. Luminal fats and bile salts appear to augment enzyme-induced bilirubin conjugation in heterotopic jejunal transplant recipients.

Problem: Mechanisms involved in infertility due to genitourinary (GU) inflammation are unknown. The production of pro-inflammatory (GRO alpha) and anti-inflammatory (IL-10) cytokines in seminal plasma is monitored in this study., Method: GRO alpha, IL-10, and granulocyte elastase were evaluated in semen from I) normal, II) infertile patients, and III) infertile patients with leukocytospermia., Results: GRO alpha in infertile patients with GU inflammation was 1.5-fold higher compared to group II and 2.5-fold higher compared to group I patients. The IL-10 was higher in group III than the other two groups. A positive correlation was observed between granulocyte elastase and GRO alpha in all groups. Group III patients exhibited poor sperm parameters., Conclusions: A shift towards increased production of pro-inflammatory chemokine GRO alpha may have a potential role in male infertility associated with GU inflammation.

Oxygen toxicity is an inherent challenge to aerobic life, including spermatozoa, the cells responsible for propagation of the species. How this toxicity affects the spermatozoan in its interactions with the ovum is still unknown. An increase in oxidative damage to sperm membranes, proteins, and DNA is associated with alterations in signal transduction mechanisms that affect fertility. Recent evidence suggests that spermatozoa and oocytes possess an inherent but limited capacity to generate ROS to aid in the fertilization process. Though a variety of defense mechanisms encompassing antioxidant enzymes (SOD, catalase, and GSH peroxidase and reductase), vitamins (E, C, and carotenoids), and biomolecules (GSH and ubiquinol) are available, a balance of the benefits and risks from ROS and antioxidants appears to be necessary for the survival and functioning of spermatozoa. An assay system for the evaluation of OSS needs to be developed. Such an assay will assist the clinician in the assessment of fertility status of both male and female partners. The determination of this OSS value will also theoretically identify the subgroups of responders and nonresponders to any putative antioxidant therapy. Though the therapeutic use of antioxidants appears attractive, clinicians need to be aware of exaggerated claims of antioxidant benefits by various commercial supplements for fertility purposes until proper multicenter clinical trial have been completed.

Objective: To quantify the levels of reactive oxygen species, superoxide dismutase (SOD), and interleukins (IL) 2 and 8 in seminal plasma of infertile patients as well as to examine the possible relationship between oxidative stress and proinflammatory cytokines., Design: Semen collected from normal fertile donors, infertile men without symptoms of genitourinary (GU) inflammation, and infertile men with symptoms of infection-inflammation of the GU tract was evaluated for the levels of granulocyte elastase, reactive oxygen species, SOD, IL-2, and IL-8. Any correlation between the levels of reactive oxygen species and other parameters in these population was analyzed statistically., Results: Significantly high levels of granulocyte elastase (18.32 +/- 1.52 U/L), reactive oxygen species (6 x 10(5) cpm), IL-8 (3.7 +/- 0.10 microgram/L), and IL-2 (18.32 +/- 1.47 ng/L) were observed in semen of infertile patients with leukocytospermia compared with the other two groups. In leukocytospermic samples, the activity of SOD was significantly lower (624.89 +/- 41.16 NU/mL) compared with nonleukocytospermic samples (787.85 +/- 24.26 NU/mL) or fertile donors (816.29 +/- 50.16 NU/mL). A significant positive correlation was observed between the levels of reactive oxygen species and IL-8., Conclusions: These findings suggest that increased oxidative stress observed during leukocytospermia may modulate the level of proinflammatory cytokines. The increased oxidative stress may be due to a defect in the reactive oxygen species scavenging system.

The generation of reactive oxygen species (ROS) and sperm damage was evaluated in (a) samples obtained during electroejaculation (EE) of men with spinal cord injury and (b) in electrolyzed HAM's F-10 medium subjected to electric current in vitro. Chemiluminescence data showed a significant increase in ROS in the ejaculates (6 x 10(7) photons/ml) collected immediately after EE and in the electrolyzed medium (3 to 7 x 10(6) photons/ml) when compared to the control (4 to 7 x 10(4) photons/ml). Incubation of normal human sperm with the electrolyzed medium resulted in a significant threefold decrease in percent motility and a twofold decrease in percent viability. Sperm subjected to direct electric stimulation in vitro exhibited a significant twofold decrease in percent motility and percent viability. Superoxide dismutase (SOD) activity decreased significantly in sperm subjected to direct electric current in comparison to the control or the sample incubated with electrolyzed medium. These studies indicate that in vitro and in vivo electrical stimulation generate reactive oxygen species and affect SOD activity, which in part are responsible for decreased sperm motion and viability.

The present study evaluates the changes in sperm motility due to H202 induced membrane damage. Washed human sperm suspended in HAM's F-10 (20-30 x 10 6/ml) were incubated (37 degrees Celsius) with varying concentrations (0-0.05%) of H202 for up to 15 minutes. Sperm were analyzed for % motility, % viability, the ratio of cholesterol to phospholipids (C/PL), and the degree of lipid peroxidation (LPO). Motility was monitored manually and viability was evaluated by the Eosin Y staining method. Total lipids were extracted with chloroform:methanol (1:2) and used in colorimetric determination of cholesterol and phospholipid contents (mcmol/106 sperm). Lipid peroxidation was measured by the production of malondialdehyde (nmol MDA/108 sperm). The results (mean +or- SEM, n=8) indicate a dose a time-dependent effect on % motility during the 15 minute incubation period. In comparison to control (8 +or- 4%), samples incubated with 0.01% H202 exhibited a 25 +or- 3% decrease in % motility, while a complete loss of motility was observed with 0.05% H202. No significant differences in decrease in sperm viability were observed between control (211 +or- 4) and H202 (0.01%) treated samples (14 +or- 2%). An increase of (54 +or- 5%) in lipid peroxidation was observed with 0.01% H202, as compared with an 18 +or- 1% increase in control samples at 15 minutes. The C/PL ratio increased by 46 +or- 4% at 15 minutes in H202 treated samples while showing a 34.3% decrease in control samples. H202 inhibited sperm motility while increasing membrane LPO and C/PL, without altering sperm viability. It would appear that lipid peroxidation and alteration of sperm membrane composition lead to the loss of sperm motility.

IDPH-791, a novel centrally acting muscle relaxant, in doses up to 500 mg/kg (po) for 14 days did not result in any appreciable adverse effect on body weight gain, food or water consumption including biochemical and haematologica parameters in rats. Variations observed in the biochemistry and haematology were either comparable to controls or were within normal limits.

The effect of calcium chloride and nifedipine on prolactin-(PRL) induced analgesia was studied by chemical assay and compared with other analgesics like morphine, thyrotrophin-releasing hormone (TRH) and clonidine. Nifedipine potentiated the analgesic effect of PRL similar to morphine while that of clonidine and TRH remained unaltered. Further, calcium chloride administration antagonized the analgesic effect of PRL and of morphine. These data suggest that PRL, similar to morphine, may alter calcium movements across the membrane to produce analgesia.