Childhood disadvantage and adolescent socioemotional wellbeing as predictors of future parenting behaviour.

INTRODUCTION: In extending work on early life antecedents of parenting, we investigate associations between childhood family history of disadvantage, adolescent socioemotional wellbeing, and age at first parenthood and subsequent parenting behaviour. METHODS: Parent-child interactions were recorded when participants in the longitudinal Dunedin Multidisciplinary Health and Development Study (New Zealand) had a three-year-old child. Data were available for 358 mothers and 321 fathers, aged between 17.7 and 41.5 at the time of their child's birth. Associations between parenting and antecedent data on socioeconomic disadvantage, adolescent wellbeing and mental health, as well as current adult mental health and age at parenting, were tested for using structural equation modelling. RESULTS: Family disadvantage in childhood and lower adolescent wellbeing was associated with less positive future parenting, but only adult (not adolescent) anxiety/depression symptoms were directly associated with parenting behaviour. Childhood family disadvantage was associated with further disadvantage across the life course that included less positive parenting of the next generation. In contrast, socioemotional wellbeing during adolescence and later age of onset of parenting were associated with more positive parenting. CONCLUSIONS: Reducing childhood disadvantage and improving socioemotional wellbeing during childhood and adolescence is likely to have intergenerational benefits through better parenting of the next generation.

Neuroprotective effects of novel phosphatidylglycerol-based phospholipids in the 6-hydroxydopamine model of Parkinson's disease.

Administration of VP025 (Vasogen Inc.), a novel drug formulation based on phospholipid nanoparticles incorporating phosphatidylglycerol, has previously been shown to have a neuroprotective effect in the brain. We examined the effect of VP025 in a rat model of Parkinson's disease, the 6-hydroxydopamine (6-OHDA) lesion of the medial forebrain bundle. VP025 or phosphate-buffered saline (PBS) was administered to rats 14 days, 13 days and 1 day before the unilateral 6-OHDA lesion. Functional integrity of nigrostriatal dopaminergic neurons was assessed 7 and 21 days later by amphetamine-induced rotational testing and we observed that rotational counts were significantly less in rats that were pretreated with VP025 compared with PBS-pretreated 6-OHDA-lesioned rats. Neurochemical analysis at 10 and 28 days after lesion revealed that VP025 protected against a 6-OHDA-induced decrease in concentrations of striatal dopamine and its metabolites. Immunocytochemical studies of the ipsilateral substantia nigra showed that VP025 significantly inhibited 6-OHDA-induced loss of dopaminergic neurons. We also observed that increases in immunostaining for activated microglia and for activated p38 in dopaminergic neurons of 6-OHDA-lesioned rats were prevented by VP025. This study shows that VP025 has significant protective effects on the 6-OHDA-lesioned nigrostriatal pathway and may therefore have potential for the treatment of Parkinson's disease.

Population pharmacokinetics of imatinib mesylate in patients with chronic-phase chronic myeloid leukaemia: results of a phase III study.

AIMS: This study was designed to investigate the biochemical and physiological covariates or comedications that affect the pharmacokinetics of imatinib mesylate in patients with chronic-phase chronic myeloid leukaemia (CP CML). METHODS: Pharmacokinetic data were analyzed in 371 patients receiving 400 mg imatinib once daily during a phase III trial of imatinib vs interferon-alfa plus cytarabine for the treatment of newly diagnosed CP CML. Covariates included age, weight, sex, ethnicity, haemoglobin (Hb) concentration, white blood cell (WBC) count, liver function, and creatinine concentration. Blood samples for imatinib analysis were taken on treatment days 1 and 29. Nonlinear mixed effects modelling was used for the population pharmacokinetic analysis. RESULTS: Population mean estimates (95% confidence interval) at day 1 for apparent clearance (CL) and apparent volume of distribution (V) of imatinib were 14 (13-15) l h(-1) and 252 (237-267) l, respectively. Modelling suggested that CL decreased by 4 (3-5) l h(-1) from day 1 to day 29, whereas V remained unchanged. Interindividual variability in CL and V was 32% and 31%, respectively. Weight, Hb, and WBC count demonstrated small effects on CL and V. Doubling body weight or Hb or halving the WBC count was associated with a 12%, 86% and 8% increase in CL, respectively, and a 32%, 60% and 5% increase in V, respectively. Comedications showed no clear effects on imatinib CL. CONCLUSIONS: Population covariates and coadministered drugs minimally affected imatinib pharmacokinetics in newly diagnosed CP CML patients.

The effect of VAS972 on allergic contact hypersensitivity.

BACKGROUND: Contact hypersensitivity (CHS) is a Th1-mediated immune response that can be down-regulated by immunosuppressive agents such as cyclosporine and environmental stimuli such as ultraviolet light. Recently, an immunomodulation therapy, VAS972, has been developed which is believed to down-regulate the Th1 arm of the immune response. This VAS972 involves modifying autologous blood by controlled exposure to the oxidizing agent ozone and UVC light, at an elevated temperature ex vivo. The processed blood is then administered by intramuscular injection. OBJECTIVE: To further evaluate the immune modulating effect of VAS972. METHODS: We examined the effect of VAS972 treatment on CHS. Contact hypersensitivity was induced with dinitrofluorobenzene (DNFB) in animals receiving VAS972- processed blood, control blood, or saline. A preliminary study was also conducted to evaluate the effect of plasma and cellular fractions of processed blood. RESULTS: Mice injected with VAS972-processed blood demonstrated a significantly lower (46%) CHS response than controls. Histologic examination of challenged ear skin from control mice displayed edema with a significant lymphocytic infiltration, whereas animals administered processed blood demonstrated a reduction in lymphocytic infiltration. Mice injected with either plasma or the cellular fraction of the VAS972-treated blood also demonstrated a significant suppression (49% and 41%, respectively). CONCLUSION: The results of this study demonstrated that VAS972 suppresses CHS and cellular infiltration. Furthermore, the plasma and cellular components of the VAS972 treatment were also able to induce immunosuppression. This further supports the hypothesis that VAS972 down-regulates the Th1 arm of the immune response.

Effect of combined heat, ozonation and ultraviolet irradiation (VasoCare) on heat shock protein expression by peripheral blood leukocyte populations.

The re-administration of whole blood subjected to heat, ozonation and ultraviolet irradiation (VasoCare therapy) has been shown to elicit clinical benefits in individuals with vascular disease. Given that these stressors induce heat shock protein (Hsp) expression and that heat shock protein reactivity is implicated in the pathogenesis of vascular disease, this study assessed the effect of VasoCare on intracellular expression of Hsp60 and Hsp70 by treated peripheral blood leukocytes. Contrary to expectations, VasoCare induced a significant reduction (approximately 40%) in the proportion of peripheral blood mononuclear cells expressing intracellular Hsp60 and Hsp70, whereas it had no effect on heat shock protein expression by peripheral blood neutrophils. Cell surface heat shock protein expression was not detectable. The reduced expression of Hsp60 by mononuclear cells was concomitant with an increase in the levels of Hsp60 in treated plasma. Although the mechanism underlying the clinical effectiveness of VasoCare therapy has yet to be established, it may be that re-administration of treated blood or soluble factors derived therefrom modifies in vivo immune responsiveness to heat shock proteins or associated molecules.

Treatment of severe Raynaud's syndrome by injection of autologous blood pretreated by heating, ozonation and exposure to ultraviolet light (H-O-U) therapy.

OBJECTIVE: To determine the effect of re-injection of small samples of autologous blood, pretreated with heat, ozone and ultraviolet light (H-O-U therapy) in patients with severe Raynaud's syndrome. EXPERIMENTAL DESIGN: Open trial in 4 patients. SETTING: Temperature/humidity controlled vascular laboratory. PATIENTS: Severe Raynaud's syndrome of more than 5 years duration and defined as more than 5 attacks daily or 10 attacks in one week, at least half of which were painful and lasting for more than 30 minutes. Three patients were refractory to infusions of Iloprost. INTERVENTIONS: Patients were treated daily or on alternate days for a two to three weeks period by re-injection of citrated autologous blood pre-treated with heat, ozone and ultraviolet light (H-O-U therapy). MEASURES: Clinical observations; mean equilibrated hand temperature (infrared thermography); distributive and microcirculatory blood-flow (venous occlusion strain-gauge plethysmography, infrared photoplethysmography, laser Doppler flowmetry) iontophoresis of acetylcholine and sodium nitroprusside; estimations: serum levels of 6-keto-PGF1alpha and serum levels of anti-hsp65 antibody. RESULTS: Reduction or abolition of Raynaud's attacks for at least three months after treatment. Mean equilibrated hand temperature increased but did not normalise. Blood flow parameters improved but did not reach statistical significance. Iontophoresis of acetylcholine showed an increase in laser Doppler flowmetry which was statistically significant. Serum levels of 6-keto-PGF1alpha, fell significantly in three patients. Serum levels of anti-hsp65 antibody fell in the one patient which was followed sequentially. CONCLUSIONS: H-O-U therapy may prove useful in patients with severe Raynaud's syndrome.

The measurement of CA 125 and placental protein 14 in uterine flushings in women with recurrent miscarriage; relation to endometrial morphology.

The concentrations of CA 125 and placental protein 14 (PP14) were measured in uterine flushings obtained throughout the luteal phase of the cycle from eight normal fertile women. The concentrations of both proteins increased in a similar pattern throughout the luteal phase of the cycle, with the most dramatic increase occurring 6 days after their luteinizing hormone surge (day LH +6). However, a greater variation in CA 125 concentrations was seen compared to that seen for PP14. The concentrations were compared to those obtained on day LH +7 of the cycle from a group (n = 35) of women with recurrent miscarriage. The ranges in concentration of PP14 and CA 125 in the flushings of fertile and recurrent miscarriage patients were very similar. However, a greater proportion of women with recurrent miscarriage (55%) had low concentrations (< 5 ng/ml) of PP14 than in the control group (12.5%) and the concentrations of PP14 in the uterine flushings were significantly less (P < 0.05) in women with recurrent miscarriage compared to the normal fertile group. There was no significant difference in the concentration of CA 125 in the uterine flushings between the two groups. Histological observation of the endometrial biopsy samples from recurrent miscarriage patients gave menstrual cycle datings that ranged from day LH +2.5 to LH +6.5 with retarded endometrium (< day LH +5) in 12 of 35 (34%) patients. Of these 12 patients, 10 (83%) had low PP14 concentrations and six (50%) had low CA 125 concentrations in their uterine flushings. In the recurrent miscarriage patients with histologically normal (> or = day LH +5) endometrial development, 10 out of 23 (43%) also had low PP14 concentrations and 8 out of 23 (35%) had low CA 125 in their uterine flushings. The results suggest that PP14 is better than CA 125 as a marker for endometrial function in this group of women. In some cases (52%) the low concentrations of PP14 in the uterine flushings could be explained by retarded endometrial development but for the others the reduction in PP14 concentration in the uterine flushing was not associated with retardation of endometrial development.

Significance of molecular marker-positive cells after autologous peripheral-blood stem-cell transplantation for non-Hodgkin's lymphoma.

PURPOSE: To evaluate the significance of molecular marker-positive cells in a cohort of non-Hodgkin's lymphoma (NHL) patients undergoing high-dose chemotherapy and autologous peripheral-blood stem-cell transplantation (PBSCT). PATIENTS AND METHODS: Twenty-eight PBSC transplants have been performed in 24 patients with poor-prognosis NHL. Molecular analysis of the t(14;18) (q32;q21) translocation (bcl-2/immunoglobulin [Ig] heavy-chain joining locus [JH] fusion) or antigen receptor gene rearrangements was performed to determine the presence of lymphoma cells at presentation, in PBSC harvests, and before and after autologous PBSCT. Kaplan-Meier estimates of survival and Cox regression analyses were used to test the effect of bone marrow involvement, tumor-cell contamination of PBSCs, disease stage, and chemotherapy sensitivity at transplantation, and presence of marker-positive cells post-PBSCT on disease-free and overall survival. RESULTS: Thirteen of 24 patients (54%) are alive following PBSCT at a median follow-up time of 654 days (range, 193 to 1,908). Nine patients are in complete remission (CR) at day 216 to 1,799 (median, 805) and four are alive following relapse (day 440, 573, 1,188, and 1,908). Eleven patients (46%) have died: three of transplant-related complications at day 0, 1, and 13, and eight of recurrent disease (day 132 to 1,330; median, 451). Longitudinal marker studies post-PBSCT showed that of 16 relapse events, 13 (81%) were positive for the lymphoma marker at or before clinically documented relapse. Marker studies became negative post-PBSCT in nine of nine patients who entered and remained in CR. Disease-free survival (DFS) was significantly shortened in patients in whom marker-positive cells were detected in serial samples posttransplantation (P = .006). Cox regression analysis showed that patients in this group had a 24 times higher risk of relapse (P = .03). CONCLUSION: The results show that the reappearance or persistence of marker-positive cells after autologous PBSCT is strongly associated with relapse.

Stimulation of human endometrial epithelial cell interleukin 6 production by interleukin 1 and placental protein 14.

The effect of interleukin 1 (IL1) and placental protein 14 (PP14) on the production of interleukin 6 (IL6) by cultured human endometrial epithelial cells prepared from endometrial biopsy material obtained at different stages in the menstrual cycle was investigated. Basal IL6 production by cells prepared from proliferative endometrium was greater than that produced by cells prepared from secretory endometrium (7.3 +/- 0.3 and 1.1 +/- 0.2 ng/well/24 h respectively, P < 0.001). IL1 (0.025-2.5 ng/ml) caused a dose-dependent increase in IL6 production by cells prepared from both proliferative and secretory endometrium, but cells prepared from secretory endometrium responded to a lower concentration of IL1 than those prepared from proliferative endometrium. IL-1-stimulated IL6 production by epithelial cells prepared from secretory endometrium typically reached 10 times basal values, while in cells prepared from proliferative endometrium stimulated levels were approximately twice the basal values. PP14 (1-50 micrograms/ml) also caused a dose-dependent increase in IL6 production by epithelial cells prepared from secretory endometrium, but had no effect on IL6 production by cells prepared from proliferative endometrium. Even in secretory cells PP14 was less effective than IL1 at stimulating IL6 production, with stimulated levels only reaching twice the basal values. This suggests that PP14 and IL1 act via different mechanisms in the stimulation of IL6 production. The results show that IL6 production by human endometrial epithelial cells is stimulated by other immunomodulatory peptides and this may be part of the network of such peptides in the endometrium which may influence embryo implantation.

The correlation of placental protein 14 concentrations in uterine flushing and endometrial morphology in the peri-implantation period.

The relationship between the concentrations of placental protein 14 (PP14) in uterine flushing and the endometrial morphology in the mid-luteal phase was assessed in a prospectively designed study involving the precise timing of all samples by the luteinizing hormone (LH) surge. A total of 29 regularly cycling women with unexplained infertility or recurrent miscarriage were studied. To flush the uterine cavity, 10 ml of physiological saline solution was used immediately prior to sampling of an endometrial specimen for morphological study, in the mid-luteal phase. PP14 concentrations were measured by radioimmunoassay in uterine flushings and plasma samples; the endometrium was assessed by the use of histological dating criteria and morphometric techniques. PP14 levels in uterine flushings were correlated with endometrial dating and volume fraction measurement of the glands. They were consistently below the sensitivity of the assay with histological dating of < day LH +5, or when the glandular lumen occupied < 20% of the gland. In contrast, PP14 concentrations in plasma were not related to histological dating or morphometric analyses, and did not differ in patients with normal endometrial development (20.8 ng/ml) and in those with retarded endometrial development (22.5 ng/ml). The presence of detectable concentrations of PP14 in uterine flushing was significantly associated with normal histological dating. Uterine flushing may therefore provide a reliable, non-invasive alternative to endometrial biopsy in the evaluation of endometrial function in the peri-implantation period.

The production of placental protein 14 and interleukin 6 by human endometrial cells in culture.

Epithelial and stromal cells prepared from endometrium taken at different times in the menstrual cycle were grown in primary culture and the production of placental protein 14 (PP14) and interleukin-6 (IL6) measured. Only the epithelial cells produced PP14. Epithelial cells from endometrium in the late secretory phase produced significantly greater amounts of PP14 (42 +/- 5.8 ng/24 h) compared with that produced by cells from early secretory and proliferative endometrium (16 +/- 1.7 and 11 +/- 1.9 ng/24 h respectively). PP14 production by cells from endometrium at all stages in the cycle was increased by progesterone, or progesterone and oestradiol together, while oestradiol alone had no effect on PP14 production. The greatest stimulation was seen during the early secretory phase when stimulated levels of PP14 reached those obtained during the late secretory phase. IL6 production by epithelial cells also varied depending on the phase of the menstrual cycle. More IL6 was produced by the cells prepared from the endometrium in the proliferative phase (10.9 +/- 0.56 ng/24 h) compared with that produced by cells from early and late secretory endometrium (2.5 +/- 0.19 and 1.45 +/- 0.09 ng/24 h respectively). Addition of steroids to the media stimulated the production of IL6 by cells from proliferative and early secretory endometrium but decreased IL6 production from cells in the late secretory phase. IL6 was also produced by stromal cells but could only be detected in supernatants of cells prepared from late secretory endometrium, and the amounts produced (0.8 +/- 0.09 ng/24 h) were less than that produced by epithelial cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Concentration of endometrial protein PP14 in uterine flushings throughout the menstrual cycle in normal, fertile women.

OBJECTIVE: To determine the variation in concentration of endometrial protein PP14 in uterine flushings throughout the menstrual cycle comparing this to concentrations in plasma samples. DESIGN: Precise timing of all samples by the luteinising hormone surge. SETTING: Jessop Hospital for Women, Sheffield. SUBJECTS: Twenty-three regularly cycling, previously fertile volunteer women. INTERVENTIONS: Observational study; 10 ml of physiological saline was used to flush the uterine cavity once or serially in the cycle of the study. MAIN OUTCOME MEASURES: The measurement of PP14 levels by radioimmunoassay in uterine flushings and plasma samples. RESULTS: In uterine flushing, PP14 levels were not detectable in significant amounts in the proliferative phase and the early luteal phase; after day LH + 6, the concentration rises rapidly with a doubling time of only 6.6 to 14.6 h in the midluteal phase. In the late luteal phase, the concentrations in uterine flushing were over a hundred times higher than the corresponding plasma samples. CONCLUSIONS: The measurement of PP14 in uterine flushings is likely to be of greater value than the measurement in plasma samples; it may provide a valuable alternative to the evaluation of endometrial function.

The effects of clomiphene citrate and cyclofenil on cervical mucus volume and receptivity over the periovulatory period.

OBJECTIVE: To determine the effects of clomiphene citrate (CC) and cyclofenil on cervical mucus (CM) volume and receptivity sampled serially over the periovulatory period. DESIGN: Using prospective luteinizing hormone (LH) timing CM volume and receptivity were compared in standard CC and cyclofenil-stimulated cycles using normal ovulatory cycles as controls. LOCATION: The Donor Insemination Unit at the University Research Clinic, Sheffield, United Kingdom. PATIENTS: Twenty anovulatory patients and 10 normally ovulating patients, all of whom were participating in a treatment cycle of donor insemination. INTERVENTIONS: The 20 anovulatory patients were allocated at random into two groups: group 1 was administered 50 mg of CC on days 2 to 6 of the menstrual cycle; group 2 was administered 400 mg of cyclofenil on days 3 to 12 of the menstrual cycle. All the patients were given a single treatment of donor insemination 24 to 36 hours after the onset of the LH surge. RESULTS: Clomiphene citrate and cyclofenil were shown to exert differential impacts on CM quantity and quality. In terms of quantity, the CC patients produced significantly lower volumes of CM than the cyclofenil patients and controls. In terms of quality, the CC patients and controls produced CM of similar receptivity, whereas the cyclofenil patients produced CM that was significantly more receptive to sperm than both the CC patients and controls. CONCLUSIONS: Neither CC nor cyclofenil exerted a detrimental impact on CM quality throughout the periovulatory period.

Round cells and sperm fertilizing capacity: the presence of immature germ cells but not seminal leukocytes are associated with reduced success of in vitro fertilization.

Fifty semen samples produced for IVF by patients diagnosed as having unexplained infertility were screened for leukocytes, leukocyte subsets, and immature germ cells using a mAB-based staining procedure. Nonfertilizing ejaculates were found to contain significantly larger numbers of immature germ cells, although no significant differences in leukocyte content were observed between groups. Neither sperm density or progressive motility were significantly different between fertilizing and nonfertilizing groups. We conclude that seminal leukocytes have little if any influence on the fertilizing capacity of the spermatozoa in patients undergoing IVF for unexplained infertility, but the presence of large numbers of germinal elements is associated with reduced fertilizing capacity and may be indicative of an immature sperm population.

The leukocytic reaction of the human uterine cervix.

PROBLEM: Sperm interaction with the immune system of the human cervix is poorly understood. METHOD: The leukocytic response of the human cervix to sperm was examined in a closely monitored patient population (N = 10), using monoclonal antibody cell identification techniques. Baseline data were collected from both cervical mucus and smears sampled before treatment by donor insemination. Donor insemination was timed to coincide with ovulation by monitoring plasma LH concentrations twice daily. Following insemination the numbers of leukocytes were recorded in cervical mucus and smear samples taken over a 24-h period relative to the time of treatment. Controls treated with "pure sperm," seminal plasma, cryopreservative, and cervical smearing alone were also included in the study. RESULTS: Only those women treated with sperm cells exhibited substantial elevations in leukocyte numbers following inseminations. Additionally, serial cervical smearing induced an inflammatory response of the cervix. In all the women, the neutrophil was the predominant leukocyte of the cervix both during the baseline and treatment periods (median values ranged from 77 to 86%). Macrophages, T-helper lymphocytes, and T-suppressor lymphocytes were also detected, but only in low numbers (2-10.6%). Two patients and one control ("pure sperm") became pregnant during their study cycle. CONCLUSIONS: We conclude that the leukocytic reaction is a physiological response of the cervix to sperm, the function of which remains to be established.

An analysis of the variation of plasma concentrations of placental protein 14 in artificial cycles.

OBJECTIVE: To analyze the factors affecting the variation of plasma concentration of placental protein 14 (PP14) in artificial cycles. DESIGN: The effects of different hormone replacement therapy (HRT) regimens were examined in a crossover design. SETTING: Jessop Hospital for Women, Sheffield, United Kingdom. PATIENTS: Eighteen women with premature ovarian failure: 6 associated with Turner's syndrome and 12 with idiopathic premature ovarian failure. INTERVENTIONS: Four different HRT regimens; 36 study cycles. MAIN OUTCOME MEASURES: Plasma PP14 concentrations on days 1, 15, 19, and 29 of the artificial cycles. RESULTS: In cycles treated with a standard HRT, the levels were similar to those of the natural cycle. Subjects with Turner's syndrome did not have elevated PP14 levels, whereas the majority (9/12 [75%]) of those with idiopathic premature ovarian failure had elevated levels on day 29 of the cycle. Levels of PP14 were reduced when either the doses of estradiol valerate were reduced to 1/3 or the doses of progesterone (P) were reduced to 1/5 of the standard HRT. CONCLUSIONS: Plasma levels of PP14 are dependent not only on P stimulation but also on adequate estrogen priming.

The removal of morphologically abnormal sperm forms by phagocytes: a positive role for seminal leukocytes?

A preliminary investigation was undertaken further to determine the function of the leukocytic cells found in semen. We performed semen analysis and quantified leukocyte subsets using immunocytochemical staining techniques in ejaculates of 351 patients. Leukocyte profiles were examined in relation to sperm morphological data for evidence of a sperm removal/selection process. Three types of seminal phagocytic cell were found to contain spermatozoa: small polymorphonuclear leukocytes (approximately 10-12 microns), monocytes of similar size and much larger (30-40 microns) macrophages capable of engulfing multiple sperm heads. The total leukocyte count (P less than 0.01), the numbers of phagocytic cells i.e. polymorphonuclear leukocytes (P less than 0.05), monocyte/macrophages (P less than 0.01) and HLA-DR positive cells (P less than 0.01), were significantly higher in those samples with greater than 50% ideal sperm forms. Significantly fewer of these same cell types were observed in samples with greater than 50% head defects. There was no difference in the number of tail or midpiece defects between leukocytospermic (greater than 10(6)/ml) and non-leukocytospermic semen samples. Oligozoospermic samples contained significantly fewer leukocytes (P less than 0.005), although above a concentration of 5 x 10(6)/ml, the sperm number was not correlated with leukocyte number. These data, along with repeated observation of spermatozoa or sperm fragments within phagocytic cells, support the hypothesis that leukocytes have a role in the removal of abnormal spermatozoa from the ejaculate.

Origin of reactive oxygen species in human semen: spermatozoa or leucocytes?

Peroxidative damage induced by reactive oxygen species (ROS) has been proposed as one of the major causes of defective sperm function. In previous studies of the production of ROS in semen, the contribution of contaminating leucocytes was not assessed. We determined the levels of ROS in 60 semen samples from men attending our infertility clinic and demonstrated by performing extraction experiments with antibody-coated magnetic beads that, within this unselected population of patients, leucocytes were the major source of ROS in the low-density Percoll fraction. Of the sperm motion parameters examined using computerized semen analysis, beat-cross frequency was the only one significantly affected by the ROS in semen.

Preliminary communication: possible role of reactive nitrogen intermediates in leucocyte-mediated sperm dysfunction.

This preliminary study was designed to examine whether nitric oxide, a reactive nitrogen intermediate produced by leukocyte metabolism of L-arginine, could reduce sperm motility. Increasing doses (10(-6)-10(-4) M) of the nitric oxide-generating drug, sodium nitroprusside (SNP) were added to motile sperm preparations and incubated over 20 hr. Quantitative sperm motility measurements were made using a computer-assisted motility analyzer in each treated sample and controls over this time period. The percentage of motile sperm, progressive motility, and concentration of motile cells were all significantly reduced with all doses of SNP. This effect was not observed in preparations treated with oxyhaemoglobin (HbO2). Mean path velocity was unaltered. Sperm viability in SNP treated sperm did not differ significantly from that of control sperm over the same time period. We conclude that further experiments are required to determine whether the production of nitric oxide/reactive nitrogen intermediates in vivo by activated leukocytes could be a contributory factor in the development of immunologic infertility.

Positive immunoselection--a method of isolating leukocytes from leukocytic reacted human cervical mucus samples.

In this brief communication we report a simple and accurate method of isolating and quantifying specific leukocytes from midcycle human cervical mucus, using monoclonal antibody-coated magnetic beads. Cervical mucus samples (pre- and postinsemination) were broken down enzymatically and incubated with a series of these beads. This method of positive immunoselection consistently retrieved representative levels of leukocytes (means = 73.8% +/- 1.59%; mean leukocyte retrieval rate +/- S.E.) from the cervical mucus samples. Significantly more leukocytes (P less than 0.0001) were isolated from the postinsemination samples, the predominant leukocyte of which was the neutrophil, which comprised 83% of the leukocyte population. These results reaffirm that a leukocytic influx is initiated across the human uterine cervix following the introduction of semen samples, the function of which is possibly phagocytic clearance of the nonfertilizing population of sperm.