Relationship between antioxidant defense in Acanthamoeba spp. infected lungs and host immunological status.

The role of oxidative stress in the pathogenicity of acanthamoebiasis is an important aspect of the intricate and complex host-parasite relationship. The aim of this experimental study was to determine oxidative stress through the assessment of lipid peroxidation product (LPO) levels and antioxidant defense mechanism in Acanthamoeba spp. lung infections in immunocompetent and immunosuppressed hosts. In Acanthamoeba spp. infected immunocompetent mice we noted a significant increase in lung lipid peroxidation products (LPO) at 8 days and 16 days post infection (dpi). There was a significant upregulation in lung LPO in immunocompetent and immunosuppressed mice infected by Acanthamoeba spp. at 16 dpi. The superoxide dismutase activity decreased significantly in lungs in immunosuppressed mice at 8 dpi. The catalase activity was significantly upregulated in lungs in immunocompetent vs. immunosuppressed group and in immunocompetent vs. control mice at 16 dpi. The glutathione reductase activity was significantly lower in immunosuppressed group vs. immunosuppressed control at 24 dpi. We found significant glutathione peroxidase downregulation in immunocompetent and immunosuppressed groups vs. controls at 8 dpi, and in immunosuppressed vs. immunosuppressed control at 16 dpi. The consequence of the inflammatory response in immunocompetent and immunosuppressed hosts in the course of experimental Acanthamoeba spp. infection was the reduction of the antioxidant capacity of the lungs resulting from changes in the activity of antioxidant enzymes. Therefore, the imbalance between oxidant and antioxidant processes may play a major role in pathology associated with Acanthamoeba pneumonia.

The inflammatory effect of infection with Hymenolepis diminuta via the increased expression and activity of COX-1 and COX-2 in the rat jejunum and colon.

The aim of this study was to determine whether Hymenolepis diminuta may affect the expression and activity of cyclooxygenase 1 (COX-1) and cyclooxygenase 2 (COX-2), resulting in the altered levels of their main products - prostaglandins (PGE2) and thromboxane B2 (TXB2). The study used the same experimental model as in our previous studies in which we had observed changes in the transepithelial ion transport, tight junctions and in the indicators of oxidative stress, in both small and large intestines of rats infected with H. diminuta. In this paper, we investigated not only the site of immediate presence of the tapeworm (jejunum), but also a distant site (colon). Inflammation related to H. diminuta infection is associated with the increased expression and activation of cyclooxygenase (COX), enzyme responsible for the synthesis of PGE2 and TXB2, local hormones contributing to the enhanced inflammatory reaction in the jejunum and colon in the infected rats. The increased COX expression and activity is probably caused by the increased levels of free radicals and the weakening of the host's antioxidant defense induced by the presence of the parasite. Our immunohistochemical analysis showed that H. diminuta infection affected not only the intensity of the immunodetection of COX but also the enzyme protein localization within intestinal epithelial cells - from the entire cytoplasm to apical/basal regions of cells, or even to the nucleus.

Hymenolepis diminuta: analysis of the expression of Toll-like receptor genes (TLR2 and TLR4) in the small and large intestines of rats. Part II.

Toll-like receptors in the gastrointestinal tract can influence intestinal homeostasis and play a role in the repair and restitution of intestinal epithelium following tissue damage. In our previous study a statistically significant increase in the level of TLR4 and TLR2 gene expression was observed in rats in early stages of hymenolepidosis. Moreover, the immunopositive cell number and the intensity of immunohistochemical staining (indicating the presence of TLRs within intestinal epithelial cells) increased over the infection period. In this paper, we determined changes in the expression of TLR2 and TLR4 and the number of anaerobic intestinal commensal bacteria in Hymenolepis diminuta infected rats. In the isolated jejunum of infected rats at 16 days post infection (dpi), the expression of TLR4 and TLR2 was significantly higher than uninfected rats. In the colon, a statistically significantly increased expression of TLR2 was observed from 16 to 40 dpi, and TLR4 from 16 to 60 dpi. The jejunum and colon of infected rats contained Gram-negative bacteria (Escherichia coli), Gram-positive bacteria (Enterococcus, Streptococcus, Staphylococcus, Bacillus, Lactobacillus) and Candida. The total number of intestinal bacteria was higher in H. diminuta infected rats, but the observed microbiota had only minor effects on the expression of TLR2 and TLR4. Toll-like receptors play a role in maintaining epithelial barrier function in response to enteric pathogens and parasites. In our study, the alteration of TLR2 and TLR4 expression in the infected rats indicates the potential role of the innate immune system in the pathomechanism of this infection.

Hymenolepis diminuta: analysis of the expression of Toll-like receptor genes (TLR2 and TLR4) in the small and large intestines of rats.

Toll receptors play a critical role in the rapid activation of innate immune responses to a variety of pathogens. In mammals, Toll-like receptors (TLR) have been found in both immune related cells and other cells. At present little is known about the participation of TLR in host defense mechanisms during parasitic infections. The aim of this study was to determine the expression of TLR2 and TLR4 genes in rat intestines during experimental hymenolepidosis. There is difference in expression of TLR2 and TLR4 genes in the colon and jejunum in uninfected rats: in the colon, mRNA of the examined TLR is present in much higher amounts than the jejunum, while the protein of the TLR also had a segmented specific distribution. In the jejunum isolated rats infected with Hymeolepis diminuta 6 and 8 days post infection (dpi), mRNA for TLR4 and TLR2 were significantly more strongly expressed in comparison with the uninfected controls. In the colon, a statistically significantly increased expression of TLR4 gene was observed only at 6 dpi, and at 8 dpi for the TLR2 gene. Moreover, we observed that during inflammation, the immunopositive cell number and the intensity of immunohistochemical staining (indicating the presence of TLR within intestinal epithelial cells), increased together with the duration of the infection period.

Seasonal dynamics of house dust mites in dust samples collected from sleeping places in north-western Poland.

The most common families of mites found in house dust are Pyroglyphidae, Glycyphagidae and Acaridae; all are a source of many antigens responsible for allergic diseases. The aim of this study was to examine the seasonal dynamics of allergenic mite populations in dust samples collected from sleeping places in apartments in north-western Poland. The mites were isolated from the dust using a saturated saline floating method. In 132 dust samples we determined: Dermatophagoides farinae, Dermatophagoides pteronyssinus, Euroglyphus maynei, Hirstia sp., Chortoglyphus arcuatus, Lepidoglyphus destructor, Gohieria fusca and Cheyletus sp. The greatest frequency was observed for D. farinae, D. pteronyssinus, Ch. arcuatus and Cheyletus sp., in the fourth quarter and D. farinae in the third quarter. Smaller coefficients of dominance were found for D. pteronyssinus, Ch. arcuatus and Cheyletus sp., and their greatest mean concentrations were found in the first and fourth quarters. Given the division of the year into heating and non-heating seasons, mites D. farinae and D. pteronyssinus achieved the highest mean concentration in the first season, and Cheyletus sp. in the second season. The analysis of the participation of developmental stages showed that the adults of D. farinae were more prevalent than juveniles in the first, second and third quarters, and imago stages of D. pteronyssinus were more numerous in relation to juveniles in the first, third and fourth quarters. The results confirm the high incidence of house dust mites in sleeping places in north-western Poland dwellings; the best conditions for the development of these mites, mainly D. farinae and D. pteronyssinus, occur in the fourth quarter and are the least favourable in the second quarter. In many cases, these results are consistent with data from other parts of Poland collected by various authors.

Electrophysiological effect of ruthenium red and pH on the skin of the edible frog, Rana esculenta L.

Amphibian skin is a sensitive interface between the organism and the environment. Metal ions from the external environment, some of them being trace elements, act on the amphibian skin. It had been shown that stimulation of tactile receptors affected Na+ transport in the frog skin and changed the potential difference, therefore the aim of this project was to study the effect of ruthenium complex, known as ruthenium red (RR), on the ion transport in this organ in vitro under control conditions, after mechanical stimulation and also in the presence of the Na+ transport inhibitor-amiloride. Three different concentrations of RR (0.12, 1.2, and 12.0 mM) in two different pH values (6.4 and 7.4) were studied in vitro in the Ussing apparatus. The measured electrophysiological parameters were the transepithelial electrical potential difference (PD) and the changes in PD after mechanical stimulation (dPD). The gentle mechanical stimulus was a jet of bath fluid from a peristaltic pump directed on the mucosal surface of isolated frog skin. After mechanical stimulation, transient hyperpolarization invariably occurred (dPD = 1.5 +/- 0.2 mV). In the presence of RR the hyperpolarization was smaller and this diminution was concentration dependent: 0.5 +/- 0.1 mV for 1.2 mM of RR and 0.1 +/- 0.1 mV for 1.2 mM of RR. At pH 6.4 the reactions of the skins on the mechanical and chemical stimuli were smaller, in the presence of amiloride disappearing completely, but after the washing away of amiloride from the experimental organ in pH 6.4 the action of RR was stimulatory. The natural defensive reactions of frog skin related to the ion transport and electrical potential difference are affected or disappear in the presence of ruthenium complex.

Pharmacological modification of ionic currents elicited in epithelia by sensory neuropeptides.

A stimulation of afferent neuronal endings in such epithelial organs as airways, colon and frog skin produces reversible hyperpolarization, usually explained as caused by neuropeptides released from sensory endings. The aim of this study was to examine pharmacological modifications of these electrophysiological reactions, which were evoked by the inhibition of Na+ or Cl- transepithelial transport by application of amiloride (AMI) and bumetanide (BUME). The tissues were mounted in a modified Ussing chamber which was additionally equipped with a nozzle connected to a peristaltic pump and acting as a stimulation device. In the resting, control conditions the transepithelial potential differences--PD of tracheal and colonic walls and frog skin were: -3.3 +/- 0.4, -3.0 +/- 0.4 and -16.5 +/- 4.1 mV, respectively and hyperpolarization after mechanical stimulation--dDP for the same organs was: -1.2 +/- 0.3, -1.2 +/- 0.3 and -2.4 +/- 0.4, respectively. Preincubation with AMI influenced the dPD in all tissues but the PD of the frog skin was only depolarized. Preincubation of the tissues with BUME diminished dPD in all organs. A hypothesis was put forward that epithelial hyperpolarization of tracheal and colonic walls depends on the combined augmentation of Na+ and Cl- transport processes, whereas epithelial hyperpolarization of frog skin depends entirely on Na+ currents and all these transepithelial ionic currents have an adapting value.