Sławomir Brzeźnicki

Predictors of environmental exposure to polycyclic aromatic hydrocarbons among pregnant women — Prospective cohort study in Poland

ObjectivesThe aim of the study was to characterize the PAH exposure level based on 1-hydroxypyrene (1-HP) in urine of Polish pregnant women and to assess the relationship between PAH and factors such as smoking, environmental tobacco smoke exposure, place of residence, heating and cooking method.Materials and MethodsThe study population included in this analysis consisted of 449 pregnant women who had been the subjects of the prospective Polish Mother and Child Cohort study performed in 8 regions of Poland. The women were interviewed three times during pregnancy (once in each trimester). 1-HP concentration in urine was chosen as the biomarker of exposure to PAH. The urine sample was analysed using high performance liquid chromatography (HPLC). The active and passive smoking exposure was verified by saliva cotinine, analysed by high performance liquid chromatography coupled with tandem mass spectrometry (LC-MSMS) and isotope dilution method.Results1-HP concentration in urine ranged from 0.02 to 10.2 μg/g creatinine with the geometric mean (GM) 0.4 μg/g creatinine. The significantly higher concentration of urinary 1-HP in pregnant women was observed for summer collection (GM ratio: 1.1; p = 0.01), among smokers (GM ratio: 1.7; p < 0.001) and for the women living in big cities (GM ratio: 1.3; p = 0.001).ConclusionsThe significantly higher concentration of urinary 1-HP in pregnant women was observed for summer collection, among smokers and those living in big cities.

EXPOSURE TO POLYCYCLIC AROMATIC HYDROCARBONS AND NEWBORN BIOMETRIC INDICATORS

OBJECTIVES The aim of the study was to examine the impact of polycyclic aromatic hydrocarbons (PAH) on foetal growth. MATERIALS AND METHODS The prospective Polish Mother and Child Cohort study was performed in 8 regions of Poland. The study population consisted of 449 mother-child pairs All women were interviewed three times during pregnancy (once in each trimester). 1-hydroxypyrene (1-HP) concentration in urine was chosen as the biomarker of exposure to PAH. The urine sample collected from the participant women between 20-24 weeks of pregnancy was analysed using high performance liquid chromatography (HPLC). The active and passive smoking exposure was verified by determination of saliva cotinine level using high performance liquid chromatography coupled with tandem mass spectrometry/positive electrospray ionisation (LC-ESI+MS/MS) and isotope dilution method. RESULTS The exposure to PAH measured by 1-HP level in urine of pregnant women was significantly associated with child birth weight (β = -158.3; p = 0.01), chest circumference (β = -0.7; p = 0.02) and cephalisation index (β = 4.2; p = 0.01) after adjustment for gestational age, child gender, pregnant woman marital status, educational level, season of last menstruation period (LMP), prepregnancy body mass index (BMI), and weight gain in pregnancy. After inclusion salivary cotinine levels into the analysis, the results were not statistically significant. CONCLUSIONS Prenatal exposures to PAH adversely influence foetal development including child weight, length, head and chest circumference. Tobacco smoking is the important source of PAH. After controlling for active and passive smoking, the observed associations were not statistically significant.

Association between a biomarker of exposure to polycyclic aromatic hydrocarbons and semen quality

ObjectivesGrowing evidence supports the reproductive and developmental toxicity of polycyclic aromatic hydrocarbons (PAHs) from prenatal and postnatal exposure, but the results of epidemiological studies regarding harmful effects of PAHs exposure on male reproductive system still remain limited and inconclusive. The aim of the present study was to investigate the relationship between 1-hydroxypyrene, a biomarker of polycyclic aromatic hydrocarbons exposure and semen quality.Materials and MethodsThe study population consisted of 277 men attending an infertility clinic for diagnostic purposes and having normal semen concentration of 20–300 mln/ml or slight oligozoospermia (semen concentration: 15–20 mln/ml) (WHO 1999). All the men were healthy and under 45 years of age. All participants were interviewed and provided a semen sample. The interview included questions concerning demographics, socio-economic status, medical history related to past diseases which may have an impact on semen quality, lifestyle factors and occupational information. Concentrations of 1-hydroxypyrene (1-OHP) in the urine samples were analyzed using high performance liquid chromatography (HPLC).ResultsA positive association was found between the level of 1-OHP in urine and sperm neck abnormalities as well as the percentage of static sperm cells (p = 0.001, p = 0.018, respectively). Additionally, exposure to PAHs measured by 1-OHP in urine decreased semen volume and the percentage of motile sperm cells (p = 0.014, p = 0.0001, respectively).ConclusionsPresented findings indicate that the environmental level of PAHs exposure adversely affects male semen quality. The future large-scale studies should incorporate different biomarkers to generate a more accurate and full assessment of the effects of PAHs exposure on male fertility.

Human sperm aneuploidy after exposure to polycyclic aromatic hydrocarbons.

The purpose of this cross-sectional study was to investigate whether environmental exposure to polycyclic aromatic hydrocarbons (PAHs) was associated with sperm aneuploidy. A sample of 181 men who attended an infertility clinic for diagnostic purposes and who had a normal semen concentration of 20-300×106 spermatozoa mL-1 or slight oligozoospermia (semen concentration of 15-20×106 spermatozoa mL-1;

Optimization of ultra-performance liquid chromatography (UPLC) with fluorescence detector (FLD) method for the quantitative determination of selected neurotransmitters in rat brain

BACKGROUND Glutamate (Glu) and γ-aminobutyric acid (GABA) are the main neurotransmitters in the central nervous system for excitatory and inhibitory processes, respectively. Monitoring these neurotransmitters is an essential tool in establishing pathological functions, among others in terms of occupational exposure to toxic substances. MATERIAL AND METHODS We present modification of the HPLC (high-performance liquid chromatography) to the UPLC (ultra-performance liquid chromatography) method for the simultaneous determination of glutamate and γ-aminobutyric acid in a single injection. The isocratic separation of these neurotransmitter derivatives was performed on Waters Acquity BEH (ethylene bridged hybrid) C18 column with particle size of 1.7 μm at 35°C using a mobile phase consisting of 0.1 M acetate buffer (pH 6.0) and methanol (60:40, v/v) at a flow rate of 0.3 ml/min. The analytes were detected with the fluorescence detector (FLD) using derivatization with o-phthaldialdehyde (OPA), resulting in excitation at 340 nm and emission at 455 nm. RESULTS Several validation parameters including linearity (0.999), accuracy (101.1%), intra-day precision (1.52-1.84%), inter-day precision (2.47-3.12%), limit of detection (5-30 ng/ml) and quantification (100 ng/ml) were examined. The developed method was also used for the determination of these neurotransmitters in homogenates of selected rat brain structures. CONCLUSIONS The presented UPLC-FLD is characterized by shorter separation time (3.5 min), which is an adaptation of the similar HPLC methods and is an alternative for more expensive references techniques such as liquid chromatography coupled with tandem mass-spectrometry (LC-MS/MS) methods. Med Pr 2017;68(5):583-591.

The capability of fungi isolated from moldy dwellings to produce toxins.

OBJECTIVES The main objective was analysis and assessment of toxinogenic capabilities of fungi isolated from moldy surfaces in residential rooms in an urban agglomeration situated far from flooded areas in moderate climate zone. MATERIAL AND METHODS The assessment of environmental exposure to mycotoxins was carried out in samples collected from moldy surfaces in form of scrapings and airborne dust from 22 moldy dwellings in winter season. In each sample 2 mycotoxins were analyzed: sterigmatocystin and roquefortine C produced by Aspergillus versicolor and Penicillium chrysogenum, respectively. Mycotoxins were analyzed by high-performance liquid chromatography (HPLC) in: scrapings from moldy surfaces, mixture of all species of fungi cultured from scrapings on microbiological medium (malt extract agar), pure cultures of Aspergillus versicolor and Penicillium chrysogenum cultured from scrapings on microbiological medium; mycotoxins in the indoor air dust were also analyzed. RESULTS The production of sterigmatocystin by individual strains of Aspergillus versicolor cultured on medium was confirmed for 8 of 13 isolated strains ranging 2.1-235.9 μg/g and production of roquefortine C by Penicillium chrysogenum for 4 of 10 strains ranging 12.9-27.6 μg/g. In 11 of 13 samples of the mixture of fungi cultured from scrapings, in which Aspergillus versicolor was found, sterigmatocystin production was at the level of 3.1-1683.2 μg/g, whereas in 3 of 10 samples in which Penicillium chrysogenum occurred, the production of roquefortine C was 0.9-618.9 μg/g. The analysis did not show in any of the tested air dust and scrapings samples the presence of analyzed mycotoxins in the amount exceeding the determination limit. CONCLUSIONS The capability of synthesis of sterigmatocystin by Aspergillus versicolor and roquefortine C by Penicillium chrysogenum growing in mixtures of fungi from scrapings and pure cultures in laboratory conditions was confirmed. The absence of mycotoxins in scrapings and air dust samples indicates an insignificant inhalatory exposure to mycotoxins among inhabitants in moldy flats of urban agglomeration situated far from flooded territories. Int J Occup Med Environ Health 2016;29(5):823-836.

P281 Exposure to widespread environmental endocrine disrupting chemicals and human sperm sex ratio

In recent years, a trend toward a declining proportion of male births has been noted in several, but not all, industrialised countries. The underlying reason for the drop in the sex ratio is unclear, but one theory states that widespread environmental endocrine disrupting chemicals affecting the male reproductive system in a negative manner could be part of the explanation. The present study was designed to investigate whether the urinary phthalate, pyrethroids and polycyclic aromatic hydrocarbons metabolites concentrations were associated with sperm Y:X ratio. The study population consisted of 194 men aged under 45 years of age who attended infertility clinic in Lodz, Poland for diagnostic purposes with normal semen concentration of 20–300 mln/ml or with slight oligozoospermia (semen concentration of 15–20 mln/ml) (WHO, 1999). The Y:X ratio was assessed by fluorescent in situ hybridization. Urinary concentrations of 1-hydroxypyrene were measured by high performance liquid chromatography, phthalate metabolites were analysed using a procedure based on the LC-MS/MS methods and metabolites of synthetic pyrethroids were assessed by gas chromatography ion-tap mass spectrometry method. After adjustment for potential confounders (past diseases, age, abstinence, smoking, alcohol consumption, sperm concentration, motility, morphology) 5 OH MEHP, CDCCA to TDCCA and 1-OHP was negatively related to Y:X sperm chromosome ratio (p = 0.033, p < 0.001, p = 0.047 respectively). As this is the first study to elucidate the association between the level of metabolites of widespread environmental endocrine disrupting chemicals (phthalates, synthetic pyrethroids, polycyclic aromatic hydrocarbons) on sex chromosome ratio in sperm therefore, these findings require further replication in other populations.