Abstract To elucidate the effects of inflammation on sperm quality, this study analysed classical sperm characteristics, leukocytes and elastase in neat semen, and sperm apoptotic markers, i.e. ...changes in plasma membrane phospholipid asymmetry, mitochondrial membrane potential (MMP), DNA integrity and intracellular reactive oxygen species (ROS), in semen prepared by density gradient using flow cytometry from 348 men of infertile couples. Increased leukocytes (⩾0.1 × 106 /ml) were associated with a decreased sperm concentration, motility and normal morphology ( P ⩽ 0.001). Sperm necrosis and DNA denaturation were increased (31.3 versus 26.6%, P = 0.020; 15.5 versus 11.5%, P = 0.011, respectively), whereas spermatozoa with normal MMP were decreased (64.1 versus 70.0%, P = 0.004). High leukocyte levels (⩾1 × 106 /ml) were not associated with any of the observed sperm parameters. At low elastase concentration (100–290 μg/l), DNA denaturation was higher (16.1 versus 10.5%, P = 0.024) compared with very low elastase concentration (<100 μg/l). A high elastase concentration (290–1000 μg/l) was associated with higher ROS index compared with low elastase concentration (1.28 versus 1.01, P = 0.016). Slightly increased leukocytes and elastase are associated with slightly poorer sperm characteristics and/or increased sperm necrosis, DNA denaturation and intracellular ROS and decreased MMP.
The authors investigated renal damage in 45 mercury miners under conditions of relatively short and low-level exposure to elemental (metallic) mercury vapour (Hg0). The analysis included urinary ...mercury, immunoelectrophoresis of urinary proteins, immunofixation and high-resolution electrophoresis, quantitative analysis of urinary albumin, and urinary alpha 1-microglobulin before and after exposure. The activity of urinary N-acetyl-beta-D-glucosaminidase (NAG) enzyme was determined after exposure. The average duration of exposure of miners was 37 (6-82) days. Urinary mercury significantly increased during exposure. Immunoelectrophoretic changes in the composition of urinary proteins occurred after exposure in 22 of 45 miners, of whom 15 showed high molecular weight (HMW) pattern of urinary proteins and seven showed low molecular weight (LMW) pattern. Only a slight increase in the urinary alpha 1-microglobulin concentration and NAG activity was found in miners with the LMW pattern of urinary proteins. The results point to a slight glomerular and tubular damage in a significant proportion of exposed miners with increased absorption of mercury vapour.
Elastase is a protease released by polymorphonuclear neutrophils (PMN) during the inflammatory process. Since 1987, seminal elastase-inhibitor complex (Ela/alpha1-PI) has been proposed as a marker of ...male silent genital tract inflammation. Measured by immunoassay in seminal plasma, Ela/alpha1-PI at a cut-off level of > or = 230 microg/l, is useful in the detection of genital tract inflammation. The prevalence of increased seminal Ela/alpha1-PI in infertile men is significantly higher than that observed in fertile men. The Ela/alpha1-PI level is positively correlated with other seminal fluid markers of male genital tract inflammation: reduced semen volume, citric acid, fructose, and increased albumin, complement component C3, caeruloplasmin, immunoglobulins IgG and IgA, and cytokines interleukins-8 and -6. A higher seminal Ela/alpha1-PI level is significantly associated with tubal damage in female partners. After antibiotic therapy, a decrease of Ela/alpha1-PI level is observed. The presence of tubal damage in the partner may negatively affect the response to antibiotic treatment. A higher seminal Ela/alpha1-PI is associated with lower percentage of sperm with single-stranded deoxyribonucleic acid (DNA) and better fertilization rate in in vitro fertilization. Besides infertility, the determination of Ela/alpha1-PI is useful to confirm the presence of prostate and other male accessory gland bacterial inflammation. Screening for PMN Ela/alpha1-PI is easy to perform and reproducible and is a reliable quantitative test for diagnosis and prognosis of silent genital tract inflammation of couples. Moreover, sequential determinations allow the follow-up of inflammation during and after therapy.
Summary
There is no consensus on whether and how male genital tract inflammation affects sperm fertilizing potential. The aims of this prospective study were to evaluate the elastase‐inhibitor ...complex in seminal plasma (s‐EI) level, a marker of male genital tract inflammation, in men undergoing in vitro fertilization (IVF) having no clinical signs of inflammation, and its association with semen characteristics, and the predictive role of s‐EI for the outcome of IVF in terms of fertilization, embryo development to the blastocyst stage and pregnancy. The study involved 104 male partners of infertile couples with normal spermiogram undergoing IVF. On the day of oocyte retrieval, spermiogram and s‐EI assessment using homogeneous immunoassay were performed. The outcome of IVF according to the s‐EI level was assessed in 85 cycles with two or more oocytes. In 67 cycles embryos were cultured to the blastocyst stage. Spearman's and Pearson's correlation tests, chi‐square test, and multiple linear and logistic regression analyses were used for statistical evaluation. Increased s‐EI level (≥250 μg/L) was detected in semen of 32% of men, and in 24% of men without leucocytospermia. We found an association between the s‐EI level and semen leucocytes (r = 0.49, p = 0.004) but not with classical sperm characteristics. No correlation between the s‐EI level and fertilization was observed. Increased s‐EI levels were associated with a poorer blastocyst development rate (p = 0.03) and a higher number of arrested embryos (p = 0.04). Extended embryo culture to the blastocyst stage shows a negative effect of clinically silent male genital tract inflammation on embryo developmental potential.
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•17% and 20% APOE ε4 carriers were identified among mothers and new-borns.•Mothers carrying APOE ε4 had higher Hg, As in venous and cord blood.•Mothers carrying APOE ε4 allele had ...higher Se in venous blood and plasma.•Statistical models confirmed maternal APOE ε4 and venous plasma Se association.•Seafood, ever-smoking, and parity had strong influence on studied associations.
We investigated the relationship between lipid binding glycoprotein apolipoprotein E (apoE; gene APOE) polymorphisms (ε4 allele carriers versus no carriers = ε4+/ε4−) and trace elements (TEs) (e.g., (methyl)mercury, arsenic, lead, cadmium, selenium, manganese, copper, and zinc) in mothers (N = 223) and their new-borns (N = 213) exposed to potentially toxic metal(loid)s from seafood consumption. The apoE isoform encoded by the ε4 allele is believed to have beneficial effects in early life but represents a risk factor for age-associated diseases. Under certain conditions ε4 carriers are more susceptible to oxidative stress and metal(loid) toxicity. DNA from Croatian pregnant women (N = 223, third trimester) and their new-borns (N = 176), was genotyped for APOE by TaqMan® SNP assay – rs429358 and rs7412. Seafood intake data and TE levels in maternal urine, milk, hair, peripheral venous blood, mixed cord blood, and new-borns’ urine were available from previous studies. We compared TEs between ε4+ and ε4− carriers using Mann-Whitney U tests and applied multiple linear regression models to analyse the TE’s dependence on the presence of allele ε4 (genotypes ε3/ε4, ε4/ε4) in combination with other explanatory variables. We identified 17% (n = 37) and 20% (n = 35) ε4 allele carriers in mothers and new-borns, respectively. The Mann-Whitney U test showed that mothers with the ε4 allele had significantly higher mean levels of (methyl)mercury in peripheral venous blood, cord blood, and hair; arsenic in urine and cord blood; and selenium in peripheral venous blood and plasma. However, taking confounders into account, only the maternal plasma selenium remained statistically significant in the linear regression models (ε4 carriers vs non-carriers: 62.6 vs 54.9 ng/mL, p < 0.001). Literature suggestions of possible ε4 allele impact on Hg levels were not observed, while superior selenium status observed in healthy pregnant women carrying allele ε4 could be linked to the proposed APOE ε4 beneficial effects early in life.
The genetic susceptibility to low-level lead (Pb) exposure in general populations has been poorly investigated and is limited to the single nucleotide polymorphism (SNP) rs1800435 in the ...delta-aminolevulinic acid dehydratase gene (ALAD). This study explored associations between ten selected ALAD SNPs with Pb concentrations in blood (BPb) and urine (UPb) among 281 men aged 18–49 years from Slovenia, including 20 individuals residing in a Pb-contaminated area. The geometric mean (range) of BPb and UPb were 19.6 (3.86–84.7) μg/L and 0.69 (0.09–3.82) μg/L SG, respectively. The possible genetic influence was assessed by examining SNP haplotypes, individual SNPs, and the combination of two SNPs using multiple linear regression analyses. While no significant associations were found for haplotypes, the presence of variant alleles of rs1800435 and rs1805312 resulted in an 11% and 13% decrease in BPb, respectively, while the presence of variant allele of rs1139488 (homozygous only) exhibited significant 20% increase in BPb, respectively. Additionally, variant allele of rs1800435 resulted in lower UPb. Individual SNPs in the model explained only around 1 additional percentage point of BPb variability. In contrast, combination analyses identified six combinations of two SNPs, which significantly explained 3–22 additional percentage points of BPb variability, with the highest explanatory power observed for the rs1800435-rs1139488 and rs1139488-rs1805313 combinations. Moreover, excluding participants from the Pb-contaminated area indicated that exposure level influenced SNPs-Pb associations.
Our results confirm the importance of the ALAD gene in Pb kinetics even at low exposure levels. Additionally, we demonstrated that identifying individuals with specific combinations of ALAD SNPs explained a larger part of Pb variability, suggesting that these combinations, pending confirmation in other populations and further evaluation through mechanistic studies, may serve as superior susceptibility biomarker in Pb exposure compared to individual SNPs.
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•ALAD SNP haplotypes were not associated with Pb levels.•rs1800435 and rs1805312 were associated with lower BPb and/or UPb.•rs1139488 was associated with higher BPb.•Specific combinations of 2 SNPs explained Pb variation better than single SNP.•Exposure level influenced associations between SNPs and Pb.
500 years of mercury (Hg) mining in the town of Idrija has caused severe pollution in Idrija and its surroundings. Following the closure of the mine in 1995, the environment remains contaminated with ...Hg. Sources of elemental-, inorganic- and methyl Hg exposure were identified, potential environmental level of exposure to Hg was evaluated and actual internal exposure to Hg was assessed in selected susceptible population groups comprising school-age children and pregnant women living in Idrija and in control groups from rural and urban environments. The study of pregnant women (n=31) was conducted between 2003 and 2008, and the study of school-age children (n=176) in 2008. Potential interaction of Hg with selenium (Se) in plasma was assessed in both study populations, while in pregnant women antioxidative enzyme activity (glutathione peroxidase, superoxide dismutase and catalase) in erythrocytes of maternal and cord blood was also assessed. Actual exposure to Hg as indicated by levels of Hg in children's blood (geometric mean (GM) 0.92µg/L), mother's blood (GM 1.86µg/L), children's urine (GM 1.08µg/g crea.), mother's urine (GM 2.51µg/L), children's hair (GM 241ng/g) and mother's hair (GM 251ng/g) was higher in the two study groups from Idrija than in the control groups from rural areas, but was still at the level of a “normal” population and reflects mainly exposure to elemental Hg (Hg°) from dental amalgam and, to a certain extent atmospheric Hg°. Furthermore, the internal doses of Hg received during pregnancy did not decrease the bioavailability of Se. Based on observation in children, the increase in Se protein expression is suggested to be a consequence of moderately elevated exposure to Hg°. The observed changes in activity of antioxidative enzymes, as biomarkers of oxidative stress, appear to be mainly associated with pregnancy per se and not with an increased exposure to Hg. In view of the continuing increased potential for Hg exposure and the low number of pregnant women studied, the results warrant a further longitudinal study of a larger group of pregnant women residing in the area of the former mercury mine.
•Estimated environmental Hg exposure in the town of Idrija still poses a potential risk to susceptible groups of inhabitants.•Actual internal exposure reflects mainly exposure to Hg° from dental amalgam and, to a certain extent atmospheric Hg°.•The internal doses of Hg received in the Idrija children and pregnant women did not decrease the bioavailability of Se.•Changes in activity of antioxidative enzymes were not associated with exposure to Hg in the town of Idrija.
The aim of the present study was to establish the population- and laboratory-specific reference intervals (RIs) for the Slovenian adult population for 24 trace elements (TEs) in blood, plasma and ...erythrocytes and to evaluate the impact of gender, age, seafood consumption, smoking habits and amalgam fillings on TEs levels.
TEs (Mn, Co, Cu, Zn, Se and Mo, Li, Be, V, Cr, Ni, Ga, As, Rb, Sr, Ag, Cd, Sn, Cs, Au, Hg, Tl, Pb and U) were determined in 192
selected blood donors (107 women and 85 men, aged 18-65 years), using inductively coupled plasma mass spectrometry (ICP-MS) with the Octopole Reaction System. Participants filled out a questionnaire, and RIs were established according to the Clinical and Laboratory Standards Institute (CLSI) guidelines for TEs.
Uniform RIs for non-essential and gender-specific for essential TEs in blood, plasma and erythrocytes were established. In our population, higher blood and plasma Cu, and erythrocyte Mn levels in women were found. In men, blood Zn, plasma Zn, Mn and Se, and erythrocyte Cu levels were higher. Zn levels were higher in 30-39 years age group. Pb and Sr increased with age. Smoking positively affected Cd, Pb, Cs and Rb; seafood consumption increased As, Hg and Zn; and amalgam increased Hg, Ag and Cu levels.
Essential TEs were inside recommended levels, and the non-essential ones were far below critical levels. Established RIs will provide an important foundation for clinical diagnostics, safety erythrocyte transfusions assessment, toxicology and epidemiological studies.
The impacts of single-nucleotide polymorphisms (SNPs) in ALAD and VDR genes on Pb health effects and/or kinetics are inconclusive at low exposure levels, while studies including APOE SNPs are rare. ...In this study, we examined the associations of ALAD, VDR and APOE SNPs with exposure biomarkers of Pb and other trace elements (TEs) in Italian pregnant women (N = 873, aged 18–44 years) and their newborns (N = 619) with low-level mixed-element exposure through diet, the environment or endogenously. DNA from maternal peripheral venous blood (mB), sampled during the second and third trimesters, was genotyped for ALAD (rs1800435, rs1805313, rs1139488, rs818708), VDR (rs2228570, rs1544410, rs7975232, rs731236) and APOE (rs429358, rs7421) using TaqMan SNP assays. Personal and lifestyle data and TE levels (mB, maternal plasma, hair and mixed umbilical cord blood CB) from the PHIME project were used. Multiple linear regression models, controlling for confounding variables, were performed to test the associations between SNPs and TEs. The geometric means of mB-Pb, mB-Hg, mB-As and mB-Cd (11.0 ng/g, 2.16 ng/g, 1.38 ng/g and 0.31 ng/g, respectively) indicated low exposure levels, whereas maternal plasma Zn and Se (0.72 μg/mL and 78.6 ng/g, respectively) indicated adequate micronutritional status. Variant alleles of ALAD rs1800435 and rs1805313 were negatively associated with mB-Pb levels, whereas a positive association was observed for rs1139488. None of the VDR SNPs or their haplotypes had any association with Pb levels. Regarding APOE, the ϵ4 allele was associated with lower mB-Hg and CB-Hg, while a positive association was found with the ϵ2 allele and CB-Pb when the model included only newborn girls. The observed associations indicate possible modification effects of ALAD and APOE SNPs on Pb or Hg kinetics in women and their newborns with low exposure to non-essential TEs, as well as an adequate nutritional status of Zn and Se.
•Maternal ALAD and APOE SNPs influenced (cord)blood Pb and Hg in pregnant women.•ALAD rs1800435 and rs1805313 variant allele was associated with lower blood Pb.•ALAD rs1139488 variant allele was associated with higher blood Pb.•APOE ε2 allele was associated with higher umbilical cord blood Pb in female newborns.•APOE ε4 allele was associated with lower blood and umbilical cord blood Hg.
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