Blood pressure in relation to biomarkers of lead, cadmium, copper, zinc, and selenium in men without occupational exposure to metals.

The interrelationship of blood lead (BPb), activity of delta-aminolevulinic acid dehydratase (ALAD), erythrocyte protoporphyrin (EP), blood cadmium (BCd), serum copper (SCu), serum zinc (SZn), serum selenium (SSe), hematocrit (Hct), body mass index (BMI), age, smoking habits, and alcohol consumption to blood pressure was examined in 154 Croatian male subjects 19-53 years of age. None of the subjects had been occupationally exposed to metals, or used any medication that could influence blood pressure or metal metabolism. The median and range values were: BPb, 57 (25-254) microg/L; ALAD, 51.9 (22.8-96.4) European units; EP, 0.68 (0.38-1.68) micromol/L erythrocytes; BCd, 0.83 (0.21-11.93) microg/L; SCu, 1113 (763-1662) microg/L; SZn, 961 (734-1213) microg/L; SSe, 73.6 (44.2-106.9) microg/L; systolic blood pressure, 131 (105-165) mm Hg; and diastolic blood pressure, 94 (71-112) mm Hg. Systolic and diastolic blood pressure significantly correlated with an increase in BMI (P<0.0005 and P<0.0001, respectively), EP (P<0.0002 and P<0.002, respectively), and BPb (P<0.005 and P<0.01, respectively). After adjusting for potential confounding variables by multiple regression, an increase in systolic blood pressure was significantly predictive by BMI (P<0.0005) and log BPb (P<0.02) and inversely by log BPb*SSe interaction term (P<0.007), or alternatively by EP (P<0.0001), BMI (P<0.001), alcohol (P<0.02), and Hct (P<0.05). An increase in diastolic blood pressure was significantly predictive by BMI (P<10(-5)), log BPb (P<0.04), and alcohol (P=0.05) and inversely by log BPb*SSe interaction term (P<0.0007), or alternatively by BMI (P<0.0001), EP (P<0.002), alcohol (P<0.004), and Hct (P<0.04) and inversely by smoking (P<0.04). With respect to the EP range in the study population, an increase of 27 mm Hg in systolic and 14 mm Hg in diastolic blood pressure was found. The study results indicate a significant Pb-related increase in blood pressure, particularly within the low-level Pb exposure range (BPb of 25-75 microg/L, and/or EP of 0.4-1.0 micromol/L erythrocytes).

Genetic markers of male infertility: Y chromosome microdeletions and cystic fibrosis transmembrane conductance gene mutations.

Today, approximately 15% of couples have reduced fertility. In most cases the reason is male infertility, usually of genetic origin. Thus, in the context of research in genes involved in reproduction and sex determination, genetic defects in gametogenesis are being extensively studied. The most frequent pathogenic causes of male infertility are Y chromosomal microdeletions and obstructive azoospermia due to congenital absence of the vas deferens (CAVD) in the presence of mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. We have investigated the most common CFTR gene alterations in Croatian men with CAVD, using Roche research prototype assays. Results revealed that the 5T variant was present in 27% of the subjects. The F508 deletion was found in 21% of the subjects. It was the most frequent mutation, although its incidence was much lower than among patients with cystic fibrosis. The prevalence of microdeletions in the azoospermia factor region (AZF) of the Y chromosome in Croatia was 4.5%. This is the first report of Y microdeletions in the Croatian population. Genetic counseling of all couples with the diagnosis of male infertility is recommended before intrauterine insemination, in vitro fertilization, and intracytoplasmic sperm injection, and should also include AZF and CFTR genotyping. Couples requesting assisted reproductive treatment should be offered molecular analysis of the CFTR gene, if male infertility due to obstructive azoospermia is the underlying cause. Also, men with severe oligozoospermia or non-obstructive azoospermia seeking assisted reproductive treatment should be screened for deletions in the Y chromosome.

Semen quality and reproductive endocrine function in relation to biomarkers of lead, cadmium, zinc, and copper in men.

Blood lead (BPb), activity of delta-aminolevulinic acid dehydratase (ALAD), erythrocyte protoporphyrin (EP), blood cadmium (BCd), serum zinc (SZn), seminal fluid zinc (SfZn), serum copper (SCu), and parameters of semen quality and of reproductive endocrine function were measured in 149 healthy male industrial workers 20-43 years of age. The group contained 98 subjects with slight to moderate occupational exposure to Pb and 51 reference subjects. All of the subjects lived in Zagreb, Croatia. Significant (p < 0.05) correlations of BPb, ALAD, and/or EP with reproductive parameters indicated a Pb-related decrease in sperm density, in counts of total, motile, and viable sperm, in the percentage and count of progressively motile sperm, in parameters of prostate secretory function (SfZn, acid phosphatase, and citric acid in seminal fluid), and an increase in abnormal sperm head morphology, serum testosterone, and estradiol. These associations were confirmed by results of multiple regression, which also showed significant (p < 0. 05) influence of BCd, SZn, SCu, smoking habits, alcohol consumption, or age on certain reproductive parameters. These effects were mainly of lower rank and intensity as compared to Pb-related reproductive effects, whereas BCd contributed to a decrease in sperm motility and an increase in abnormal sperm morphology and serum testosterone. No significant Pb- or Cd-related influence was found on levels of the lactate dehydrogenase isoenzyme LDH-C(4) and fructose in seminal fluid or on follicle-stimulating hormone, luteinizing hormone, and prolactin in serum. The seminal fluid concentrations of Pb (SfPb) and Cd (SfCd) were measured in 118 of the 149 subjects, and a highly significant (p < 0.0001) correlation was found between BPb and SfPb levels (r = 0.571) and between BCd and SfCd levels (r = 0.490). The overall study results indicate that even moderate exposures to Pb (BPb < 400 microg/L) and Cd (BCd < 10 microg/L) can significantly reduce human semen quality without conclusive evidence of impairment of male reproductive endocrine function.

Cadmium in the blood and seminal fluid of nonoccupationally exposed adult male subjects with regard to smoking habits.

Blood cadmium (B-Cd) and seminal fluid cadmium (Sf-Cd) were measured in 120 adult male subjects not occupationally exposed to cadmium (Cd), comprising 42 nonsmokers (including nine former smokers) and 78 smokers. The respective median and range values were: 0.46 (0.19-1.49) microgram/l of B-Cd and 0.54 (0.17-1.67) microgram/l of Sf-Cd in nonsmokers, and 4.33 (0.49-13.33) micrograms/l of B-Cd and 0.85 (0.29-3.56) microgram/l of Sf-Cd in smokers. Both indicators showed a highly significant difference in Cd exposure between the groups (P < 0.0001), although the increase in B-Cd was considerably more pronounced than that of Sf-Cd in smokers compared with nonsmokers. The results suggest a nonlinear relationship (log Sf-Cd/log B-Cd: r = 0.501, P < 0.0001), rather than linear relationship (Sf-Cd/B-Cd: r = 0.430, P < 0.0001), between the indicators. Significant correlations were found between smoking habits, i.e., the number of cigarettes per day, and an increase in B-Cd in smokers (r = 0.296, P < 0.01) and in all 120 subjects (r = 0.685, P < 0.0001), as well as between smoking habits and an increase in Sf-Cd in smokers (r = 0.378, P < 0.001) and in all 120 subjects (r = 0.488, P < 0.0001). Both indicators are necessary for evaluation of individual internal Cd dose, since they appear to differ in reflecting recent and long-term cumulative Cd exposure and/or the amount of Cd at the site(s) of its effect(s) in the body.

Sperm motility and kinetics of dynein ATPase in astheno- and normozoospermic samples after stimulation with adenosine and its analogues.

We tested the effects of adenosine and 2-deoxyadenosine on the activation of human spermatozoa. In the asthenozoospermic group of patients adenosine produces an increase in sperm motility from 33.3 +/- 2.1% to 42.1 +/- 3.4%, progressive motility from 22.5 +/- 1.3% to 28.6 +/- 1.7% and forward progression rating from 2.1 +/- 0.2% to 2.8 +/- 0.1%. 2-Deoxyadenosine stimulated asthenozoospermic samples to a greater degree than adenosine. Sperm motility rose to 48.9 +/- 3.4%, progressive motility to 32.1 +/- 3.4% and forward progression rating to 3.0 +/- 0.1% following stimulation with 2-deoxy-adenosine. The kinetic parameters and basic characteristics of dynein ATPase were determined. The maximum activity of dynein ATPase, Vmax, was significantly different (P < 0.001) for asthenozoospermic and normozoospermic samples: 6.46 +/- 2.1 nmol Pi/mg/min and 16.99 +/- 3.7 nmol Pi/mg/min respectively. However, the enzyme affinity for ATP was not different. Stimulation of asthenozoospermic samples with adenosine and 2-deoxyadenosine caused an increase of Vmax (70-90% and 90-110% respectively) and no significant change in KM was observed. In order to block the nucleoside transporter and to eliminate the action of adenosine inside the cell, dipyridamole was used but the effects of adenosine were not neutralized. 5'-(N-ethylcarboxy-amido)-adenosine showed effects similar to those of adenosine, even when applied in 1 microM concentration. These results indicate that adenosine and its analogues stimulate sperm motility and activity of dynein ATPase, most probably via A2 receptors.

Retrograde ejaculation and loss of emission: possibilities of conservative treatment.

Twenty-five patients with retrograde ejaculation/loss of emission were treated with ephedrine sulfate or imipramine hydrochloride. Seventeen of them suffering from both diabetes and retrograde ejaculation were treated with ephedrine or, in case that ephedrine failed to convert retrograde ejaculation into anterograde ejaculation, with imipramine. Positive results were obtained in 5/17 (29.3%) patients, i.e. in 3 (17.6%) and 2 (11.7%) patients on ephedrine and imipramine, respectively. The daily dose of ephedrine was 50 mg and that of imipramine 75 mg, during a 4-week period. In the group with retroperitoneal lymphadenectomy, after treatment with ephedrine, only 1 (12.5%) had retrograde ejaculation while the remaining patients (n = 7) continued to lack semen emission. These 7 patients were treated with imipramine, and 3 of them (42.8%) achieved anterograde ejaculation. In one third of patients with retroperitoneal lymphadenectomy and diabetes, with retrograde ejaculation or loss of semen emission, conservative treatment can offer improvement or conversion to anterograde ejaculation.

Seminal plasma LDH-X in testosterone rebound therapy.

The changes in seminal plasma LDH-X isoenzyme were investigated in 27 oligozoospermic patients treated with large doses of testosterone. The increase in this isoenzyme, proven to occur prior to the sperm count elevation, might possibly serve as a predictor enabling the choice of an optimal term for affecting conception. A high percentage of responders has been found in patients with severe oligozoospermia (less than 0.1 x 10(6) spermatozoa/ml), but with a high initial relative percentage of seminal plasma LDH-X. The determination of this parameter could be one of the criteria for the selection of patients who will probably respond to the therapy.

Semen LDH-X deficiency and male infertility.

Sperm-specific isozyme LDH-X (LDH-C4) in the semen of some infertile men with a normal sperm concentration was absent. This finding suggests a need for semen LDH-X isozyme determination in the evaluation of some causes of male infertility.

Seminal plasma isoenzyme LDH X in infertile men.

The seminal plasma isoenzyme LDH X and hormonal profile were determined on 55 samples from patients with eigher azoo- or oligozoospermia. The correlation found between the value fo LDH X and the gonadotropins can be used as an indicator of germ cell differentiation. Surprisingly high values of LDH X which were found in patients with severe oligozoospermia could indicate that the patients have normal spermatogenesis and possibly may respond to hormonal therapy.

Decreased insulin binding to erythrocytes in subjects with Klinefelter's syndrome.

Insulin binding to specific erythrocyte receptors was investigated in group of 25 subjects with Klinefelter's syndrome (47 XXY genotype) and 14 healthy male volunteers. Insulin binding was significantly decreased in Klinefelter subjects (P less than 0.01 at insulin concentrations of 0.051 and 0.136 mmol/liter); however, their fasting glucose concentration was normal (87 +/- 17), and the glucose disappearance rate was slightly increased (2.3 +/- 0.9; P less than 0.2). These data indicated a compensatory, mechanism involved in the glucose metabolism in Klinefelter's syndrome.