Johan Juhl Weisser

Sediment baseline study of levels and sources of polycyclic aromatic hydrocarbons and heavy metals in Lake Nicaragua

Selected metals and polycyclic aromatic hydrocarbons (PAHs) were analyzed in sediment samples from 24 sites in Lake Nicaragua sampled May 2010 to provide a baseline of pollution levels. Cu exceeded the Consensus-Based Sediment Quality Guideline (CBSQG) Threshold Effect Concentrations (TECs) at 21 sites while Ni exceeded the value at one site. Comparison of the sampling sites showed that the south-eastern shore and a central part of the lake contained the highest levels of As, Cd, Cr, and Ni, while the western part of the lake contained the highest levels of Cu, Pb, and Zn. Analysis of PAH levels showed that the CBSQG TECs were exceeded by naphthalene at five sites. The sum concentrations of the 16 US EPA priority PAHs (∑PAH16) ranged from 0.01 mg kg(-1) dw to 0.64 mg kg(-1) dw. The highest ∑PAH16 concentration was found upstream in River Mayales and the PAH composition revealed a heavy PAH fraction (e.g., creosote). The main sources of PAHs in Lake Nicaragua were determined as of diffuse petrogenic and pyrogenic origin as well as diagenetic produced perylene. The relative importance of these PAH sources was determined by interpretation of loading and score plots from a principal component analysis. This study concluded that areas of Lake Nicaragua represent an important pollution baseline for future studies in this lake and other tropical lakes.

Enantioselective endocrine disrupting effects of omeprazole studied in the H295R cell assay and by molecular modeling

Enantiomers possess different pharmacokinetic and pharmacodynamic properties and this may not only influence the therapeutic effect of a drug but also its toxicological effects. In the present work we investigated the potential enantioselective endocrine disrupting effects of omeprazole (OME) and its two enantiomers on the human steroidogenesis using the H295R cell line. Differences in production of 16 steroid hormones were analyzed using LC-MS/MS. Additionally, to evaluate the differences in binding modes of these enantiomers, docking and molecular dynamics (MD) simulations of S-omeprazole (S-OME) and R-omeprazole (R-OME) in CYP17A1, CYP19A1 and CYP21A2 were carried out. Exposing H295R cells to OME and its enantiomers resulted in an increase of progesterone (PRO) and 17α-hydroxy-progesterone (OH-PRO) levels. At the same time, a decrease in the corticosteroid and androgen synthesis was observed, indicating inhibition of CYP21A2 and CYP17A1. In both cases, the effect of R-OME was smaller compared to that of the S-OME and a certain degree of enantioselectivity of CYP17A1 and CYP21A2 was suggested. Docking indicated that the N-containing rings of OME possibly could interact with the iron atom of the heme for S-OME in CYP17A1 and S- and R-OME in CYP21A2. However, density functional theory calculations suggest that the direct N-Fe interaction is weak. The study demonstrates enantioselective differences in the endocrine disrupting potential of chiral drugs such as omeprazole. These findings may have potential implications for drug safety and drug design.

Developing a new research tool for use in free-ranging cetaceans: recovering cortisol from harbour porpoise skin

In this paper we present a new analytical method that allows the extraction of the stress hormone cortisol from naturally sloughed cetacean skin cells. Skin cortisol is unaffected by short-term cortisol fluctuations and could thus be used to assess chronic stress levels in cetaceans.

Simvastatin decreases steroid production in the H295R cell line and decreases steroids and FSH in female rats.

Endocrine modulating effects of Simvastatin (SV) and its metabolite, Simvastatin β-hydroxy acid (SVA), were investigated in H295R cells and in female Sprague-Dawley (SPRD) rats. H295R cells were exposed to SV and SVA concentrations from 0 to 10μM for 48h. Four groups of SPRD rats received 0 (CT), 1.3 (L), 5.0 (M), and 20.0 (H)mg SV/kg bw/day for 14 days. 10 Steroids were investigated in H295R growth media, and in tissues and plasma from rats using GC-MS/MS. Plasma LH and FSH were quantified by ELISA. In the H295R assay, SV and SVA particularly decreased progestagens with IC50-values from 0.10-0.13μM for SV and from 0.019-0.055μM for SVA. In rats, SV decreased progestagens in ovaries, brain and plasma, and plasma FSH in the M (72.4% decrease) and H group (76.6% decrease). Because progestagens and gonadotropins are major players in fertility, administration of SV might exert negative effects on female reproduction.

A novel method for analysing key corticosteroids in polar bear (Ursus maritimus) hair using liquid chromatography tandem mass spectrometry

This paper presents the development and evaluation of a methodology for extraction, clean-up and analysis of three key corticosteroids (aldosterone, cortisol and corticosterone) in polar bear hair. Such a methodology can be used to monitor stress biomarkers in polar bears and may provide as a useful tool for long-term and retrospective information. We developed a combined pressurized liquid extraction (PLE)-solid phase extraction (SPE) procedure for corticosteroid extraction and clean-up followed by high pressure liquid chromatography tandem mass spectrometry (HPLC-MS/MS) analysis. This procedure allows for the simultaneous determination of multiple steroids, which is in contrast to previous polar bear studies based on ELISA techniques. Absolute method recoveries were 81%, 75% and 60% for cortisol, corticosterone and aldosterone, respectively. We applied the developed method on a hair sample pooled from four East Greenland polar bears. Herein cortisol and corticosterone were successfully determined in levels of 0.32±0.02ng/g hair and 0.13±0.02ng/g hair, respectively. Aldosterone was below limit of detection (LOD<0.17ng/g). The cortisol hair concentration found in these East Greenland polar bears was consistent with cortisol levels previously determined in the Southern Hudson Bay and James Bay in Canada using ELISA kits.

Occurrence and risk assessment of antibiotics in water and lettuce in Ghana

Hospital wastewater and effluents from waste stabilization ponds in Kumasi, Ghana, are directly discharged as low quality water into nearby streams which are eventually used to irrigate vegetables. The presence of 12 commonly used antibiotics in Ghana (metronidazole, ciprofloxacin, erythromycin, trimethoprim, ampicillin, cefuroxime, sulfamethoxazole, amoxicillin, tetracycline, oxytetracycline, chlortetracycline and doxycycline) were investigated in water and lettuce samples collected in three different areas in Kumasi, Ghana. The water samples were from hospital wastewater, wastewater stabilization ponds, rivers and irrigation water, while the lettuce samples were from vegetable farms and market vendors. Antibiotics in water samples were extracted using SPE while antibiotics in lettuce samples were extracted using accelerated solvent extraction followed by SPE. All extracted antibiotics samples were analyzed by HPLC-MS/MS. All studied compounds were detected in concentrations significantly higher (p=0.01) in hospital wastewater than in the other water sources. The highest concentration found in the present study was 15μg/L for ciprofloxacin in hospital wastewater. Irrigation water samples analyzed had concentrations of antibiotics up to 0.2μg/L. Wastewater stabilization ponds are low technology but effective means of removing antibiotics with removal efficiency up to 95% recorded in this study. However, some chemicals are still found in levels indicating medium to high risk of antibiotics resistance development in the environment. The total concentrations of antibiotics detected in edible lettuce tissues from vegetable farms and vegetable sellers at the markets were in the range of 12.0-104 and 11.0-41.4ng/kg (fresh weight) respectively. The antibiotics found with high concentrations in all the samples were sulfamethoxazole, erythromycin, ciprofloxacin, cefuroxime and trimethoprim. Furthermore, our study confirms the presence of seven antibiotics in lettuce from irrigation farms and markets, suggesting an indirect exposure of humans to antibiotics through vegetable consumption and drinking water in Ghana. However, estimated daily intake for a standard 60kg woman was 0.3ng/day, indicating low risk for human health.

Sertraline Suppresses Testis and Adrenal Steroid Production and Steroidogenic Gene Expression While Increasing LH in Plasma of Male Rats Resulting in Compensatory Hypogonadism

Selective serotonin reuptake inhibitors are used as first line treatment in major depressive disorder. However, selective serotonin reuptake inhibitors have also been associated with sexual disorders, abnormalities, and sexual dysfunction, although mechanisms are unclear. The aim of this project was to investigate the possible endocrine disrupting effect of sertraline (SER) on sex steroid production in male rats exposed to 3 therapeutically realistic doses of SER 1.25, 5, and 20 mg/kg/day. To achieve this, we analyzed all the major steroids in testis, adrenals, brain, and plasma using Liquid chromatography tandem mass spectrometry. Furthermore, we investigated the potential effects on gene expression on the major genes involved in testicular, adrenal and brain steroidogenesis using quantitative PCR. Moreover, plasma luteinizing hormone (LH) levels were analyzed. We observed significant reduction in steroid production, in particular on the testicular Δ-4 axis and on the adrenal CYP17-hydroxylase axis. Effects in brain and plasma were less pronounced. Testicular gene transcription was also significantly down-regulated except for the CYP19 (aromatase) gene. In contrast, no effects on the adrenal gene expression were observed, except for an up-regulation of the CYP17. Plasma LH and LH/TS were increased, in particular in the lowest exposure group, indicating some degree of compensatory hypogonadism. In conclusion, this study demonstrates extensive endocrine disruption during SER exposure in male rats, both directly on steroid production in major endocrine tissues, but also indirectly by affecting gene expression. Furthermore, increased LH levels may augment decreased sex steroid production, in particular testosterone production, inducing a state of compensatory hypogonadism.

Occurrence, removal and risk assessment of steroid hormones in two wastewater stabilization pond systems in Morogoro, Tanzania

The present study investigated the occurrence and removal of 10 steroid hormones (4 androgens, 3 progestagens and 3 estrogens) in two WSP systems, Mafisa and Mzumbe in Morogoro, Tanzania. All 10 steroid hormones were detected in the influent of both WSP systems in the dry as well as in the rainy season. The concentrations of steroids in influent wastewater ranged from 0.1 ng/L for 17-OH-pregnenolone to 445 ng/L for estrone and from below limit of detection for 17-OH-pregnenolone to 45 ng/L for estrone in effluent. During dry season, the overall mean ± standard deviation removal efficiency for the 10 steroids were 70 ± 21% for Mzumbe WSP and 97 ± 3% for Mafisa WSP. During the rainy season the overall mean removal efficiency for all the steroid hormones were 52 ± 32% for Mzumbe WSP and 94 ± 8% for Mafisa WSP. Risk was characterized by calculating the risk quotients (RQs) for fish and humans. 46% of the total RQs calculated were above one, indicating high risk. Low RQs were estimated for androgens and progestagens but the estrogen concentrations measured in the WSP systems and Morogoro River indicated a high risk for fish. However, estrogens appeared not to pose an appreciable risk to human health from water intake and fish consumption. The results indicated that WSP systems are quite effective in removing steroid hormones from wastewater. Thus, low technology systems such as WSP systems are suitable techniques in low income counties due to relatively low costs of building, operating and maintaining these systems.

The Combination of Fosfomycin, Metronidazole, and Recombinant Human Granulocyte-Macrophage Colony-Stimulating Factor is Stable in vitro and Has Maintained Antibacterial Activity

BACKGROUND Treatment of secondary peritonitis includes surgery and antimicrobial agents. Antimicrobial agents are often administered intravenously, however, the alternative route intraperitoneal administration could be considered. Investigations must be conducted prior to clinical application. Therefore, we aimed to investigate the combination of fosfomycin, metronidazole, and recombinant human granulocyte-macrophage colony-stimulating factor with regard to its chemical properties and the solutions stability. In addition, the antibacterial effect of the mixed drug solution was compared with the effect of the individual antibacterial agents. METHODS The drugs were mixed to an aqueous solution. Basic chemical investigations of pH, precipitation, and calculated osmolarity of the drug combination were conducted. Fosfomycin and metronidazoles chemical stability was investigated using High Pressure Liquid Chromatography-Mass Spectrometry. Microbiological investigations using the agar cup method were carried out to measure the antibacterial effect of fosfomycin and metronidazole. RESULTS The aqueous solution of the combination of the three drugs had a pH of 7.46-7.62, which was stable during 24 h, was without precipitation, and had a calculated osmolarity of 293 mOsm/l. High Pressure Liquid Chromatography-Mass Spectrometry found stable concentrations of fosfomycin and metronidazole both alone and in combination during 24 h. The antibacterial effect of the drug combination solution was similar to the antibacterial effects of fosfomycin and metronidazole alone. CONCLUSION The drug combination had neutral and stable pH, was iso-osmotic, and had stable concentrations during 24 h of storage. The antibacterial effect of fosfomycin and metronidazole were not altered when the drugs were mixed.