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D Buckingham

Publications and source records attributed to D Buckingham.

13 recordsLinked to original sources
PubMed2000

Impact of a deep saturation dive on semen quality.

The demonstration dive 'Aurora' has provided an opportunity to study the impact of extreme hyperbaric conditions on male fertility. This operation involved a 33-day diving programme during which divers were exposed to a maximum pressure of 4.6 Mega Pascals (Mpa) for 7 days. At days - 4, + 27, + 34, + 82 and + 263 relative to the initiation of the dive, semen samples were analysed to determine the quality of spermatogenesis and the functional competence of the spermatozoa. A dramatic fall in semen quality was observed in association with the dive and by day + 82 the potential fertility of the men was seriously compromised as evidenced by oligoasthenoteratozoospermic semen profiles and the poor fertilizing potential of the spermatozoa. These studies indicate, for the first time, that the severe hyperbaric conditions associated with deep saturation dives have a profound effect on male reproductive function.

Diving↗
PubMed1994

Relationships between biochemical markers for residual sperm cytoplasm, reactive oxygen species generation, and the presence of leukocytes and precursor germ cells in human sperm suspensions.

In this study, we have examined the relationship between creatine phosphokinase (CPK), a biochemical measure of human sperm quality (Huszar et al., 1988a,b, 1990; Huszar and Vigue, 1994), and a marker for the presence of residual cytoplasm in human spermatozoa, glucose-6-phosphate dehydrogenase (G6PDH). We then determined whether the diagnostic potential of these enzymes was related to the capacity of the sperm suspensions to generate reactive oxygen species (ROS) and/or the presence of leukocytes and precursor germ cells. Across the data set as a whole, G6PDH and CPK were highly correlated with each other and, to a lesser extent, with the generation of ROS. Contamination of the sperm suspensions with leukocytes might have contributed to these associations, since the presence of such cells was also significantly correlated with CPK, G6PDH, and ROS. However, even after the leukocytes had been carefully removed, G6PDH was still highly correlated with CPK (r = 0.794), indicating that both criteria were providing similar information of the cytosolic component of human sperm suspensions. In the absence of leukocyte contamination, CPK and G6PDH activities were also correlated with the presence of precursor germ cells, and this association may, in part, explain the diagnostic value of these criteria. An additional component of their prognostic value may be reflected in the statistically significant association observed between G6PDH activity and ROS generation. A possible mechanism for such an association is suggested, which should be amenable to experimental verification.

Biomarkers↗
PubMed1994

Analysis of the extent to which sperm movement can predict the results of ionophore-enhanced functional assays of the acrosome reaction and sperm-oocyte fusion.

This study has examined the extent to which the information generated by ionophore-enhanced bioassays of the acrosome reaction and sperm-oocyte fusion might be predicted from the computer-aided analysis of sperm motility. Strong correlations (r approximately 0.7) were observed between specific components of sperm movement in semen and the potential for A23187-induced sperm-oocyte fusion, generating a stepwise regression coefficient of R = 0.663 on the bais of two criteria, percentage progressive motility and amplitude of sperm lateral head displacement (ALH). The movement characteristics of the spermatozoa recovered from the Percoll gradients gave an even higher R value of 0.838 on the basis of four variables (percentage rapid, average path velocity, straightness and ALH). In contrast, the ability of human spermatozoa to undergo acrosome reaction in response to A23187 exhibited a limited correlation with sperm movement, whether these measurements were made in the original semen sample or following Percoll purification (R approximately 0.4). These results have diagnostic implications, since sperm-oocyte fusion and the acrosome reaction clearly differ in their relative dependence on sperm motility. In practical terms, it should be noted that the computer-aided analysis of sperm movement was shown to provide up to 70% of the information generated by the more laboured assessment of sperm-oocyte fusion.

Acrosome↗
PubMed1993

New TS treatment.

Explore the source record for details and available documents.

1-Naphthylamine↗
PubMed1993

Use of a xanthine oxidase free radical generating system to investigate the cytotoxic effects of reactive oxygen species on human spermatozoa.

The reaction between xanthine and xanthine oxidase results in the univalent and divalent reduction of dioxygen to generate superoxide (O2-.) and hydrogen peroxide (H2O2), respectively. With the aid of this system, the direct effect of reactive oxygen species (ROS) on human sperm function has been investigated. A protocol involving the addition of xanthine oxidase to the reaction mixture at 0 and 15 min resulted in a loss of motility involving every component of sperm movement examined. Lower doses of xanthine oxidase, which did not influence sperm motility, were also found to suppress the competence of human spermatozoa to exhibit oocyte fusion in response to the ionophore, A23187. The reactive oxygen species responsible for the disruption of human sperm function was not influenced by the presence of superoxide dismutase (SOD) or scavengers of hypochlorous acid or hydroxyl radicals. However, the cytotoxic species was shown to be extremely stable and could be completely eliminated by catalase, which selectively eliminates H2O2. Confirmation that it is H2O2, and not O2-., which is cytotoxic to human spermatozoa was obtained in studies in which the direct addition of this oxidant was shown to influence both the movement of human spermatozoa and their competence for oocyte fusion. These results carry implications for the diagnosis of defective sperm function and the design of optimized culture media for the treatment of male factor infertility.

Calcimycin↗
PubMed1993

Relationship between iron-catalysed lipid peroxidation potential and human sperm function.

The relationship between lipid peroxidation and the functional competence of human spermatozoa has been investigated in a cohort of 31 infertility patients. Lipid peroxidation was assessed using a sensitive fluorometric assay for the generation of malondialdehyde in response to the presence of a ferrous ion promoter. Sperm function was evaluated by monitoring the movement characteristics of these cells and their capacity for sperm-oocyte fusion. Each sample was separated into high- and low-density sperm populations on discontinuous, two-step (40%:80%), Percoll gradients prior to analysis. The way in which individual ejaculates fractionated on these gradients was highly positively correlated (P < 0.001) with the lipoperoxidation status of the spermatozoa; the greater the potential for malondialdehyde generation, the higher the proportion of cells entering the low density region of the gradients. The lipoperoxidation potential of the freshly prepared spermatozoa was also highly predictive (P = 0.0001) of their capacity for movement at 3 and 24 h and their ability to exhibit sperm-oocyte fusion in response to the ionophore A23187. The potential for malondialdehyde generation in the 40% and 80% Percoll fractions was positively associated with midpiece abnormalities in the spermatozoa. These results emphasize the importance of lipid peroxidation in the pathophysiology of male infertility and suggest a mechanism by which such damage might arise.

Female↗
PubMed1992

Development of a technique for monitoring the contamination of human semen samples with leukocytes.

OBJECTIVE: To determine whether receptors for the N-formyl chemotactic peptide, FMLP, exist on the surface of human spermatozoa and regulate reactive oxygen species generation by these cells. DESIGN: Chemiluminescent analysis of reactive oxygen species generation by suspensions of human spermatozoa before and after removal of leukocytes using a magnetic cell separation technique. SETTING: Academic Research Institute. PATIENTS: Unselected male volunteers. RESULTS: Human sperm suspensions responded to FMLP and phorbol ester (PMA) with a burst of reactive oxygen species production. Autoradiographic analyses employing 3H FMLP and chemiluminescence studies involving the selective removal of leucocytes with anti-CD 45-coated magnetic beads demonstrated that the FMLP responses were because of leukocyte contamination. In contrast, reactive oxygen species production in response to PMA appeared to reflect the oxidant-generating capacity of both leukocytes and spermatozoa. CONCLUSION: The only cells present in the human ejaculate possessing detectable receptors for FMLP and capable of generating reactive oxygen species in response to this reagent come from the leukocyte population. Luminol-dependent, FMLP-induced, chemiluminescence provides a rational basis for monitoring the presence of leukocytes in suspensions of human spermatozoa.

Cell Fractionation↗
PubMed1992

Enhanced detection of reactive oxygen species produced by human spermatozoa with 7-dimethyl amino-naphthalin-1, 2-dicarbonic acid hydrazide.

A new chemiluminescence technique has been assessed for the detection of reactive oxygen species generated by purified populations of human sperm. This revised protocol involves the use of horse-radish peroxidase (HRP) in combination with a luminol analogue, 7-dimethyl amino-naphthalin-1,2-dicarbonic acid hydrazide (DNDH), that exhibits two-three times the quantal efficiency of luminol itself. The chemiluminescent signal generated with these reagents was significantly (P less than 0.001) greater than that obtained with the conventional luminol-based methodology for both the steady-state situation and following stimulation of the sperm with PMA and A23187. Dose-response analyses indicated that the DNDH/HRP chemiluminescence system could give linear standard curves with hydrogen peroxide concentrations into the nmol l-1 range. In contrast, the exponential rise in chemiluminescence recorded with luminol was not observed until hydrogen peroxide concentrations exceeded 10 mumol l-1. It is concluded that the enhanced sensitivity of the DNDH/HRP system to low levels of hydrogen peroxide should facilitate the application of chemiluminescent techniques to the diagnosis of oxidative stress in cases of male infertility.

Fluorescent Dyes↗
PubMed1992

Differential contribution of leucocytes and spermatozoa to the generation of reactive oxygen species in the ejaculates of oligozoospermic patients and fertile donors.

Cells isolated from the ejaculates of a high proportion of patients exhibiting oligozoospermia are characterized by generation rates of reactive oxygen species that considerably exceed those obtained for the normal fertile population. The purpose of this study was to resolve the cellular source of this enhanced activity. Semen samples from a cohort of oligozoospermic patients and a group of fertile controls were fractionated on discontinuous Percoll gradients to generate three cell populations (0, 50 and 100%) of differing density. For each fraction, both the steady-state and the phorbol-ester-induced chemiluminescent signals were significantly (P less than 0.001) greater for the oligozoospermic samples than for the fertile controls. In the fertile donors, leucocytes comprised the major source of reactive oxygen species, particularly in the low-density Percoll fractions; in oligozoospermic patients, however, spermatozoa were identified as a second major source of reactive oxygen species. Particularly striking was an intense phorbol-ester-induced chemiluminescent signal generated by oligozoospermic spermatozoa, purified by passage through isotonic Percoll and free of leucocyte contamination, which was 167 times greater than the median signal generated by the corresponding fraction from the fertile controls (P less than 0.001). These results emphasize the importance of spermatozoa as a major source of reactive oxygen species in oligozoospermia and have implications for the diagnosis and treatment of this condition, as well as for the design of appropriate diagnostic strategies.

Cell Separation↗
PubMedDate not supplied

Sperm penetration into a hyaluronic acid polymer as a means of monitoring functional competence.

The diagnostic significance of sperm penetration assays based on a commercially available hyaluronate preparation (Sperm Select) has been investigated in the male partners of 77 couples characterized by a normal female partner. Sperm penetration into hyaluronate was highly correlated with the ability of the same sperm populations to penetrate bovine cervical mucus and, moreover, depended on the same attributes of semen quality, including the morphology of the spermatozoa, their number, and their motility as reflected by their mean path velocity. Stepwise multiple regression analyses employing these independent variables generated r values of 0.821 to 0.931, depending on the criterion of hyaluronate penetration used; path velocity was consistently the most informative variable according to the standardized regression coefficients. The relationship between hyaluronate penetration and sperm movement was so close that multiple regression equations could be generated that were capable of accounting for up to 76% of the variance in sperm velocity measurements obtained with a computerized image analysis system. Regression equations could also be generated using the hyaluronate penetration data that could account for 65% of the variance observed in an A23187-enhanced zona-free hamster oocyte penetration test, including the successful identification of the subpopulation of patients in whom 0% oocyte penetration had been recorded. Within the same data set, independent variables based on bovine cervical mucus penetration could only account for 43.5% of the variance in sperm-oocyte fusion. Hyaluronate penetration therefore appears to offer a simple, objective means of generating information on the functional competence of human spermatozoa that should find a role in routine diagnostic services where the more specialized tests are not available.

Animals↗
PubMedDate not supplied

Leukocytic infiltration into the human ejaculate and its association with semen quality, oxidative stress, and sperm function.

Immunocytochemical techniques have been used to monitor the size and composition of the leukocyte population in unfractionated human semen samples and sperm populations generated by Percoll gradient centrifugation. The characteristics of the leukocyte population have then been related to the quality of the semen profile, the production of reactive oxygen species, and the functional competence of the spermatozoa. A majority (97%) of the ejaculates examined contained leukocytes, and in 82.4% the major cell type was the granulocyte. Small numbers of T cells, B cells, and monocytes/macrophages could also be found in 62%, 43%, and 21% of samples, respectively, and patients were occasionally identified in whom one of these cell types became the predominant leukocyte species. Although a subpopulation of patients was identified in whom the infiltration of multiple leukocyte species was positively correlated with the concentrations of spermatozoa and precursor germ cells in semen, in general, the presence of leukocytes, to the point of leukocytospermia, did not significantly influence any component of the semen profile. Similarly, the fertilizing potential of the washed spermatozoa, as assessed by in vitro tests of the acrosome reaction and sperm-oocyte fusion, was not correlated with the concentration of seminal leukocytes. In contrast, the carryover of leukocytes into the washed sperm preparations profoundly influenced the fertilizing potential of the spermatozoa via mechanisms that were associated with the production of reactive oxygen species. These results have implications for the diagnostic significance of leukocyte contamination in the context of male infertility and assisted conception.

Cell Movement↗