Morphometric changes in the corpus luteum of pregnant hypokinetic rats

The aim of this work was to study the influence of hypokinetic conditions on the ovary and corpus luteum of pregnant rats. The rats were kept in hypokinetic conditions for 5 days in the period between the 13th and 18th days of pregnancy. A three-dimensional reconstruction of the ovary and corpora lutea and also a stereological evaluation of the luteal cells and their nuclei were performed using serially cut material. Hypokinesia caused a decrease in the mean volume of the ovary and individual corpus luteum and in the total volume of corpora lutea per ovary in immobilised animals as compared to the control. Moreover, a decrease was observed in the mean number of luteal cells and an increase in the size of these cells, as well as in the mean volume fraction of their nuclei. These results indicate that immobilisation of pregnant rats for 5 days considerably influences the morphology of the corpus luteum and luteal cells

Endogenous Kynurenine Aminotransferases Inhibitor is Proposed to Act as “Glia Depressing Factor” (GDF)

The endogenous neuroinhibitory amino acid receptor antagonist kynurenic acid (KYNA) has been hypothetically linked to physiological processes and to the pathogenesis of several brain disorders. The aim of this study was to search KYNA metabolism i.e. KYNA levels and enzymes synthesising KYNA kynurenine aminotransferase I and II (KAT I and II) in the central nervous system (CNS) and in the peripheral nervous system. Within the investigated species we found a remarkably low KYNA content (3.4 nM) in piglet’s serum compared to rat and human serum. Furthermore, in contrast to high KAT activity present in rat and human livers, a lack of KAT I and KAT II activity was found in piglet liver and other piglet peripheral organs. Therefore we attempted to find a reason for the absence of KYNA formation in piglet peripheral tissue and we researched to find if KYNA formation in rat liver homogenate (measured under standard assay conditions for KAT activity) can be influenced by the application of piglet tissue homogenates and other body fluids. KYNA formation in rat liver homogenate was investigated in the presence of piglet liver, piglet brain, rat brain and human brain homogenates, and also in the presence of cerebrospinal fluid (CSF) of the control and of Multiple Sclerosis patients. We found a significant and dose dependent reduction of rat liver KAT I and KAT II activities in the presence of piglet brain, piglet liver, and human brain, but not in the presence of rat brain homogenate. Interestingly, CSF of the human control subjects significantly lowered rat liver KAT I activity. Furthermore, the inhibitory effect of CSF of Multiple Sclerosis (MS) patients was significantly weaker when compared to the CSF of control subjects. Our data, for the first time, indicated the presence of active component(s)—depressing factor—in the body, which was able to block KYNA formation. Reduced KAT inhibitory effect by CSF of MS patients would suggest a lowered “depressing factor” level in CSF of MS patients and is possibly responsible for an enhancement of KYNA formation and for glia activation and gliosis in the CNS. Subsequently, two fractions obtained after centrifugation of CSF from patients with Neuroborreliosis showed a significantly different ability to block KAT I activity. The CSF-sediment fraction exerts a stronger inhibitory activity than the CSF-supernatant fraction, supporting further the presence of a depressing factor. For the first time, data revealed and demonstrated the ability of endogenous components to block KYNA’s synthesis. We propose that a glia depressing factor (GDF), which is abundantly present in the body, might simultaneously control glia cell’s KAT activity, respectively KYNA synthesis and also glia proliferation. The mechanism(s) of action, the composition and structure of this factor needs to be further elaborated

Proliferation of macrophages due to the inhibition of inducible nitric oxide synthesis by oxidized low-density lipoproteins

Oxidized low-density lipoprotein (ox–LDL) is assumed to be a major causal agent in hypercholesteraemia-induced atherosclerosis. Because the proliferation of lipid-loaden macrophages within atherosclerotic lesions has been described, we investigated the dependence of macrophage proliferation on the inhibition of inducible nitric oxide synthase (iNOS) by hypochlorite oxidized LDL. Ox–LDL induces a dose dependent inhibition of inducible nitric oxide synthesis in lipopolysaccharide-interferon stimulated mouse macrophages (J774.A1) with concomitant macrophage proliferation as assayed by cell counting, tritiated-thymidine incorporation and measurement of cell protein. Native LDL did not influence macrophage proliferation and inducible nitric oxide synthesis. iNOS protein and mRNA was reduced by HOCl-oxidized LDL (0–40 µg/ml) as revealed by immunoblotting and competitive semiquantitative PCR. Macrophage proliferation was increased by the addition of the iNOS inhibitor L–NAME. The addition of ox–LDL to L–NAME containing incubations induced no further statistically significant increase in cell number. Nitric oxide donors decreased ox–LDL induced macrophage proliferation and nitric oxide scavengers restored macrophage proliferation to the initial values achieved by ox–LDL. The decrease of cytosolic DNA fragments in stimulated macrophages incubated with ox-LDL demonstrates that the proliferative actions of ox-LDL are associated with a decrease of NO-induced apoptosis. Our data show that inhibition of iNOS dependent nitric oxide production caused by hypochlorite oxidized LDL enhances macrophage proliferation. This might be a key event in the pathogenesis of atherosclerotic lesions

Zagrożenia zdrowotne wśród dzieci i młodzieży. T. 2

Praca recenzowana / Peer-reviewed pape

Effects of Various Kynurenine Metabolites on Respiratory Parameters of Rat Brain, Liver and Heart Mitochondria

Previously, we demonstrated that the endogenous glutamate receptor antagonist kynurenic acid dose-dependently and significantly affected rat heart mitochondria. Now we have investigated the effects of L-tryptophan, L-kynurenine, 3-hydroxykynurenine and kynurenic, anthranilic, 3-hydroxyanthranilic, xanthurenic and quinolinic acids on respiratory parameters (ie, state 2, state 3), respiratory control index (RC) and ADP/oxygen ratio in brain, liver and heart mitochondria of adult rats. Mitochondria were incubated with glutamate/malate (5 mM) or succinate (10 mM) and in the presence of L-tryptophan metabolites (1 mM) or in the absence, as control. Kynurenic and anthranilic acids significantly reduced RC values of heart mitochondria in the presence of glutamate/malate. Xanthurenic acid significantly reduced RC values of brain mitochondria in the presence of glutamate/malate. Furthermore, 3-hydroxykynurenine and 3-hydroxyanthranilic acid decreased RC values of brain, liver and heart mitochondria using glutamate/malate. In the presence of succinate, 3-hydroxykynurenine and 3-hydroxyanthranilic acid affected RC values of brain mitochondria, whereas in liver and heart mitochondria only 3-hydroxykynurenine lowered RC values significantly. Furthermore, lowered ADP/oxygen ratios were observed in brain mitochondria in the presence of succinate with 3-hydroxykynurenine and 3-hydroxyanthranilic acid, and to a lesser extent with glutamate/malate. In addition, 3-hydroxyanthranilic acid significantly lowered the ADP/oxygen ratio in heart mitochondria exposed to glutamate/malate, while in the liver mitochondria only a mild reduction was found. Tests of the influence of L-tryptophan and its metabolites on complex I in liver mitochondria showed that only 3-hydroxykynurenine, 3-hydroxyanthranilic acid and L-kynurenine led to a significant acceleration of NADH-driven complex I activities. The data indicate that L-tryptophan metabolites had different effects on brain, liver and heart mitochondria. Alterations of L-tryptophan metabolism might have an impact on the bioenergetic activities of brain, liver and/or heart mitochondria and might be involved in the development of clinical symptoms such as cardiomyopathy, hepatopathy and dementia

Spatial distribution and risk assessment of heavy metals in bottom sediments of two small dam reservoirs (south-east Poland)

Sediments of two dam reservoirs in SE Poland, Zalew Zemborzycki (ZZ) and Brody Iłżeckie (BI) were studied. The sediments from both reservoirs were sampled in the transects perpendicular to the shoreline, at the river inflow and the frontal dam. The total concentration of Mn, Zn, Pb, Cd, Cu, Cr and Ni was determined by ICP-EAS method after the sample digestion in the mixture of concentrated HNO3 and HClO4 acids. The statistical analyses: value intervals, mean values, variation coefficient, the median and the skewed distribution were performed. To estimate differences between the means for transects, Tukey’s test was applied with least significant difference (LSD) determination. The maps of the metal spatial distribution were drawn and sediment quality according to the geochemical and ecotoxicological criteria evaluated. Differences between the reservoirs in terms of heavy metals concentration in bottom sediments, and regularities in their spatial distribution were found. In the ZZ sediments the concentration was at the level of geochemical background (Zn, Cr), slightly (Cd, Cu, Ni) or moderately (Pb) contaminated sediments. The metal concentration in the sediments of the BI was up to eight times higher as compared to the ZZ. Moreover, sediments from the BI reservoir showed a greater variability of metal concentration than those from ZZ, which resulted from the dredging operation performed in the part of the reservoir. Metal concentration in sediments of the dredged part was ca. 2–5 times lower than in the undredged one, which indicates that after the dredging operation, accumulation of these metals was slight. The concentrations of Zn, Pb and Cd from the undredged part of BI were at the level of contaminated sediments and exceeded the probable effects level (PEL). In the ZZ, the greatest accumulation of metals occurred in the upper part of the reservoir and at the frontal dam, and the lowest in the middle part of the reservoir. In BI, the lower outflow of water in this reservoir caused a lower metal concentration in the sediments at the frontal dam, as compared with the other sediments in the undredged part of the reservoir. The results indicate that in small and shallow reservoirs, areas of accumulation of heavy metals depend on such factors as a parent river current, reservoir depth, water waving, reservoir shape (narrowing, coves/bays), and type of water outflow