The effects of prometryne on subchronically treated mice evaluated by SCGE assay.

Prometryne is a methylthio-s-triazine herbicide used to control annual broadleaf and grass weeds in many cultivated plants. Significant traces are documented in environment, mainly water, soil and plants used for human and domestic animal nutrition. Data on the toxic effects of prometryne and other methylthio-s-triazine have scorcely been published. The goal of this study was to investigate if prometryne, applied orally, could induce DNA damage in mouse leukocytes, in subchronical in vivo experimental design. Three different doses of prometryne were applied per os repeatedly every 48 hours. After the 7th dose (day 14) and the 14th dose (day 28) blood leucocytes were analyzed by alkaline Single Cell Gel Electrophoresis (Comet) assay. The results of three different comet parameters showed general increase in Olive tail moment, tail length and tail intensity values in treated groups of animals. The increase in measured values was almost proportional to the dose received and the time of exposure. We conclude that prometryne or its metabolic residues have the potential to induce processes that cause genotoxic effects on leukocytes on mice in in vivo repeated exposure.

Effects of prometryne on apoptosis and necrosis in thymus, lymph node and spleen in mice.

Prometryne is a methylthio-s-triazine herbicide. Significant traces are documented in environment, mainly waters, soil and plants used for nutrition. The aim of this study was to estimate prometryne immunotoxic properties through induction of apoptotic and/or necrotic changes in thymocytes, splenocytes and lymph node cells after repeated subchronical exposure. Three different doses of prometryne (185, 375, 555mgkg(-1)) were applied per os every 48h, over 28 days. Flow cytometry assay (annexinV-FITC and PI) was conducted to record apoptotic and necrotic damage. In the spleen significant changes in the percentage of apoptotic cells were not detected between treated and control groups respectively. In thymus and lymph node, within the lowest dose group (185mgkg(-)1), an increase in percentage of early apoptosis without any significant increase in necrosis was detected. Medium (375mgkg(-1)) as well as high dose triggered increase in late apoptosis in lymph node while in thymus; late apoptosis was increased only in animals exposed to the highest dose (555mgkg(-1)). The highest applied dose, in thymus and lymph node respectively, caused a general decrease in percentage of vital cells in favour of marked increase of percentages of all types of dying cells (apoptotic, late apoptotic/early necrotic and necrotic). Prometryne caused disbalance in major organs of immune system, markedly lymph nodes and thymus, by induction of early apoptotic changes in dose/time specific manner.

Ovarian follicular concentration of IL-12, IL-15, IL-18 and p40 subunit of IL-12 and IL-23.

BACKGROUND: The aim of the study was to determine the presence of interleukin (IL)-12, IL-15, IL-18 and p40 subunit of IL-12/IL-23 in follicular fluid from spontaneous cycles and the relation between the concentration of selected cytokines and IVF-embryo transfer outcome. METHODS: IVF-embryo transfer and enzyme immunoassay (EIA) (R&D Systems, Minneapolis, MN, USA and MBL, Nagoya, Japan) were used. RESULTS: Follicular fluid of women included in the IVF-embryo transfer procedure contained common p40 subunit of IL-12/IL-23 (median 70.1 pg/ml), IL-15 (median 1.3 pg/ml) and IL-18 (median 38.2 pg/ml). There was a significant negative correlation between follicular fluid concentrations of IL-15 and IL-18 (R=-0.392, P=0.003). Significantly higher concentrations of common p40 subunit of IL-12/IL-23 (median 79.8 pg/ml) were found in the follicular fluid taken from follicles containing oocytes, when compared with those without an oocyte (median 44.5 pg/ml, P=0.006). Patients who achieved clinical pregnancy had significantly decreased concentration of IL-15 (median 0.8 pg/ml) compared with patients without successful IVF-embryo transfer outcome (median 1.4 pg/ml, P=0.047). CONCLUSION: Follicular fluid collected from spontaneous cycles contains detectable levels of p40 subunit of IL-12/IL-23, IL-15 and IL-18. Increased concentrations of p40 subunit of IL-12/IL-23 in follicles containing oocytes suggest an important role of this cytokine in reproduction. Possible negative value of IL-15 as a predictor of IVF-embryo transfer success remains to be determined.

Persistant immunodominant anti-gag SLYNTVATL responses in HIV-patients with up to 7 years of HAART.

We analyzed Gag-specific CD8+ T-cells in HIV-patients on long-term HAART and in untreated chronically-infected patients by using iTAg MHC class I tetramers (HLA-A*0201) specific for SLYNTVATL. Gag SLYNTVATL-specific CD8+ T-cells were detectable in 18 of 26 treated patients (median 5.2 years of HAART) and in 10 of 14 untreated patients. Median percentage of Gag SLYNTVATL-specific CD8+ T-cells in treated patients was 0.10 (range 0.00-0.70%). Median number of Gag SLYNTVATL-specific CD8+ T-cells per 50,000 CD8+ T-cells was 56.0 cells (range 2.0-344.0 cells) and was not significantly different compared with untreated patients (p = 0.978). Numbers of Gag SLYNTVATL-specific CD8+ T-cells were inversely correlated with the duration of undetectable plasma viremia (p = 0.02, Rho = -0.430). Chronically-infected HIV-patients on HAART (for up to 7.7 years) maintained a stable subpopulation of Gag SLYNTVATL-specific CD8+ T-cells. This finding is relevant for the analysis of treatment-induced immune reconstitution and, possibly, for future therapeutic strategies in HIV-disease.

Increased expression of CXCR3 and CCR5 on memory CD4+ T-cells migrating into the cerebrospinal fluid of patients with neuroborreliosis: the role of CXCL10 and CXCL11.

The aim of this study was to evaluate the contribution of chemokine receptor CXCR3 and the corresponding ligands CXCL10 and CXCL11 to the recruitment of peripheral blood (PB) memory CD4+ T-cells into the cerebrospinal fluid (CSF) of patients with acute neuroborreliosis. Percentages of memory CD45RO+CD4+ T-cells expressing CXCR3 and CCR5 were significantly enriched in the CSF compared to the PB. Concentrations of CXCL10 and CXCL11 in the CSF of neuroborreliosis patients were significantly higher compared with the corresponding serum samples. Our results suggest that CXCL10 and CXCL11 create a chemokine gradient between the CSF and serum and recruite CXCR3-expressing memory CD4+ T-cells into the CSF of neuroborreliosis patients and that CCR5 also plays a role in this process.