N-acetyl-beta-D-Glucosaminidase in Tissues of Rats Chronically Exposed to Cadmium and Treated with Ozone.

Our aims were to evaluate N-acetyl-beta-D-glucosaminidase (NAG) activity in an experimental rat model of chronic exposure to cadmium and its response to ozone therapy. Forty male Wistar rats were divided into five groups: control, cadmium only, cadmium and oxygen, cadmium and ozone, and ozone only. Cadmium concentration (ASA method) and NAG activity (by the Maruhn method) were determined in the supernatants of the kidneys, liver, and pancreas. The histopathological alterations were evaluated in tissue sections.The highest concentration of cadmium and NAG activity was observed in rats intoxicated with cadmium. Ozone therapy led to a decrease in cadmium accumulation in the kidneys and liver. An examination of renal, hepatic and pancreatic tissues revealed severe histopathological lesions in Cadmium group (Cd) treated animals. The histopathological changes in animals treated with ozone were similar, but with slightly decreased intensity. Positive correlations between histochemical lesions, NAG activity and cadmium concentration in the study groups were observed. It has been shown that chronic cadmium intoxication has cytotoxic activity in the kidneys, liver, and pancreas, causing an increase in NAG activity. Ozone therapy significantly reduces NAG activity and the severity of histopathological lesions in the kidneys and liver, confirming its beneficial effects.

The Effects of Duodenojejunal Omega Switch in Combination with High-Fat Diet and Control Diet on Incretins, Body Weight, and Glucose Tolerance in Sprague-Dawley Rats.

BACKGROUND: Despite excellent results of bariatric surgery in the treatment of type 2 diabetes and weight loss in human subjects, some patients do not obtain desired results. One of the reasons for this is that not all patients follow caloric intake recommendations. AIM: The aim of this study was to investigate the effect of duodenojejunal omega switch (DJOS) surgery on body weight, glucose tolerance, and incretins in rats. METHODS: DJOS and SHAM surgery were performed on rats maintained for 8 weeks on high-fat diet (HF) and control diet (CD), respectively. After surgery, four groups were kept on the same diet as before the surgery, and four groups had a changed diet (CD vs. HF and HF vs. CD) for the next 8 weeks. Glucose tolerance, insulin tolerance, glucose-stimulated insulin, glucagon-like peptide-1 (GLP-1) and gastric inhibitory polypeptide/glucose-dependent insulinotropic polypeptide (GIP) secretion, food intake, and body weight were measured. RESULTS: A change of diet after surgery resulted in reduced glucose tolerance. Plasma insulin levels were lowered between DJOS and SHAM surgeries for the HF/HF and CD/HF groups. DJOS surgery did not reduce body weight in the studied groups, irrespective of diet. In the HF/HF group, ΔGLP-1 was lower for DJOS surgery in comparison with other groups. Differences of weight changes were observed for groups HF/HF and HF/CD. After DJOS surgery, ΔGIP was lower in the CD/HF group compared with HF/HF. CONCLUSIONS: Our results show that applications of different types of diets, before and after surgery, is a sensitive method for studies of mechanism of glucose intolerance after DJOS surgery.

Long-term Effect of Ileal Transposition on Adipokine Serum Level in Zucker (Orl)-Lepr(fa) Fatty Rats.

BACKGROUND: Ileal transposition surgery is an increasingly used procedure in combination with sleeve gastrectomy to control obesity and type 2 diabetes mellitus (T2DM). A short-term effect on glycemia amelioration after the ileal transposition (IT) procedure is observed; however, it appears that the effect is time dependent, and it remains uncertain if this effect is also linked with an adipose tissue hormonal activity. METHODS: Twenty male Zucker rats underwent IT or sham surgery. Six months after surgery, serum levels of adiponectin, vaspin, resistin, chemerin RBP4 were analyzed using ELISA kits. Tissue concentrations of glycogen sythase kinase alfa (GSK-3α), glucose 6-phosphatase (G6PC), glycogen phosphorylase (PYGM), and phosphofructokinase (PFK) in muscle and GLUT4 in visceral fat, white adipose tissue, and muscle were assessed in duplicate by an enzyme-linked immunosorbent assay (ELISA) kit. Additionally, the transposed ileum and analogical ileal segment of sham-operated rats were processed for histomorphometry analysis. RESULTS: The animals which underwent IT showed significantly a higher adiponectin and vaspin serum level. Concentrations of resistin decreased after IT surgery but were not significantly different between the groups. The plasma level of chemerin decreased significantly after IT and correlated negatively with adiponectin serum level in the IT group. The effects of IT on RBP4 serum level appeared to be significantly lower than those in the sham group and correlated with GLUT4 concentration in IT white adipose tissue negatively, but positively with the sham group. CONCLUSIONS: The data suggest that ileum transposition leads to a stimulatory effect on important adipokines involved in glucose metabolism. The adipokine serum level could be a useful biomarker of postoperative physiological state.

DNA damage in grasshoppers' larvae--comet assay in environmental approach.

The comet assay that provides a quantitative measure of the DNA-strand breaks may be used for assessing the 'genotoxic potential' of the environment. Young adults of Chorthippus brunneus (Orthoptera), collected at three sites in Southern Poland, differing in the level of pollution, particularly with heavy metals: Pilica (reference), Olkusz (moderately polluted) and Szopienice (heavily polluted) - were allowed to mate under laboratory conditions that were free from any pollution. Egg-pods were collected and, after diapause, brain cells from one-day old larvae were used for the comet assay. We compared the level of DNA damage in the larvae originating from these sites and also measured time-dependent DNA repair after single 10min. application of H2O2 (20µM final concentration). The DNA damage was relatively low in larval cells irrespectively of the site pollution their parents came from. However, measured comet parameters - tail DNA content (TDNA), tail length (TL), and olive tail moment (OTM) - were significantly higher in larvae originating from the Szopienice site than in those from the reference site. Incubation of cells with H2O2 resulted in significantly higher values of the comet parameters in the insects from all the study sites with the highest ones observed in the offspring of grasshoppers from Szopienice. Moreover, DNA repair, following the treatment, did not occur in the latter group. These data contribute to almost unexplored subject of genotoxic effects of environmental pollutants in insects. They are discussed in the light of the concept of adaptive strategies in energy allocation depending on the level of biotope pollution.

Digestive enzymes activity in subsequent generations of Cameraria ohridella larvae harvested from horse chestnut trees after treatment with imidacloprid.

In the present study we describe the effect of chloronicotinoid pesticide (imidacloprid) on the digestive enzymes activity of the Cameraria ohridella larvae after lasting 1 year sublethal exposure to imidacloprid pesticide. Caterpillars - L4 stage (fourth instar, hyperphagic tissue-feeding phase) - were collected from chemically protected white horse chestnut trees 1 year after imidacloprid treatment, and compared with caterpillars collected from non-treated trees in a previous study. Enzymes activity of α-amylase, disaccharidases, glycosidases and proteases was assayed. The presence of pesticide in ingested food changed the digestive enzymes profile of caterpillars. The analysis of correlations between different digestive enzymes showed many significant correlations (P<0.05) among glycolytic activities like ß-glucosidase and α-galactosidase activities. Statistically significant correlations for proteolytic activity were found between trypsin and chymotrypsin activity and aminopeptidase activity that occurred only in the 1st generation. PCA distinguished five primary components with eigenvalues higher than 1, from which the first two explain almost 59% of analyzed results. Surprisingly, in the pesticide treated groups significantly higher activities of sucrase and lactase in relation to control were found. In general, glycosidase (α-glucosidase, ß-glucosidase and ß-galactosidase) activities showed a similar pattern of activity in different generations. These results contrast with those obtained with control larvae, where significant differences in activities of α-glucosidase, ß-glucosidase and ß-galactosidase may result from the different quantity and quality food intake by subsequent generations of larvae. No inter-generation differences in total proteolytic activity were observed in treated larvae. The absolute value of total proteolytic activity was higher than that in the control group. The pesticide present in the vascular system of the horse chestnut tree significantly affected some of the digestive enzymes activities and - in consequence - also interrelationships between enzymes, what may affect the food digestion.

Within and between seasonal changes of detoxifying capabilities of Cameraria ohridella (Lepidoptera : Gracillariidae) larvae.

Cameraria ohridella is an invasive leaf miner, a severe pest of horse chestnut trees. Chemical control needs recognition of intrinsic metabolic capability to cope with external stressing factors. Our tasks were to check annual effects of generation, and host tree age on detoxifying abilities of the last larval stage. Activities of CAT, SOD, GSTPX, GST CarE and AChE were assayed in the midgut of larvae from two localities during 3 years and three generations, annually. Activities of GSTPX and GST were high, but CAT and SOD were low in relation to other lepidopteran larvae. In general, the second generation larvae had the highest activity, indicating an effective defence against allelochemicals produced by the host. Effects of host tree age were significant for SOD, GSTPX and CarE activities. Significant annual differences on enzyme profiles of CAT, SOD, CarE and AChE reflected differences of temperature and precipitation between consecutive years.

Alteration of carbohydrates metabolism and midgut glucose absorption in Gromphadorhina portentosa after subchronic exposure to imidacloprid and fenitrothion.

This study was undertaken to test the hypothesis that following exposure to insecticides, changes take place in the metabolism of carbohydrates and absorption in the midgut of insects. The Madagascar hissing cockroach (Gromphadorhina portentosa) was chosen for the experiment as a model organism, due to it being easy to breed and its relatively large alimentary tract, which was important when preparing the microperfusion midgut bioassay. In each group of cockroaches treated with imidacloprid and fenitrothion, absorption of glucose, expressed as the area under the curve (AUC), was elevated compared to the control group. Glucose in the hemolymph of the examined insects was present in a vestigial amount, often below the threshold of determination, so the determinable carbohydrate indices were: hemolymph trehalose concentration and fat body glycogen content. The level of trehalose found in the hemolymph of insects when exposed to fenitrothion, and irrespective of the level of concentration mixed into food, were significantly lower when comparing to the control samples. Imidacloprid acted analogically with one exception at the concentration of 10 mg·kg(-1) dry food where trehalose concentration did not differ from the control values. Coupling with fat body glycogen concentration was less visible and appeared only at the concentrations of 5 and 10 mg imidacloprid·kg(-1) dry food. As described in this study changes in the sugar distribution and midgut glucose absorption indicate that insects cover the increased energy needs induced by insecticides; also at the gastrointestinal tract level. The result indicates that the midgut glucose absorption parameters could be considered as a non-specific biomarker of insecticide toxicity.

Digestive enzymes activity in larvae of Cameraria ohridella (Lepidoptera: Gracillariidae).

This article presents the activity of carbohydratases and proteases in the midgut of Cameraria ohridella larvae--an oligophagous pest whose preferred feeding is horse chestnuts leaves. Optimal media pH of the assayed enzymes were similar to those of other Lepidopterans. Relatively high amylase activity, as well as maltase and sucrase activities, indicates that starch and sucrose are the main digested saccharides. Trehalase activity was similar to that described in other Lepidopterans. Activities of glycosidases were significantly lower than those of disaccharidases what suggests that neither cellulose nor glycosides are important for C. ohridella. Trypsin is the main endoprotease of this pest. Like in other leaf-eaters carboxypeptidase activity was higher than that of aminopeptidase. The activity of the majority of examined enzymes increased in the following successive pest generations, which could be explained by the decreased nutritional value of older leaves. Probably this phenomenon in hydrolases activity in Cameraria is a nonspecific mechanism present at this stage of co-evolution of the horse chestnut and its pest.

Cellular stress reactions assessed by gender and species in spiders from areas variously polluted with heavy metals.

In the funnel web spider Agelena labyrinthica (Agelenidae; A. l.), sheet web spider Linyphia triangularis (Linyphiidae; L. t.) and wolf spider Xerolycosa nemoralis (Lycosidae; X. n.) from two differently polluted meadow sites in southern Poland, we studied the relations between antioxidant parameters (glutathione, GSH; glutathione peroxidases, GPOX, GSTPx; catalase, CAT; stress proteins-Hsp70, metallothioneins Mts), the intensity of apoptosis and necrosis, and heavy metal burdens of the midgut gland. Cellular reactions against stress caused by pollutants seemed to be sex-dependent. The concentrations of Zn and Cu in the midgut glands of male A. l. and X. n. were more than double that of the females, from both study sites. In male spiders from the heavily polluted site, both negative correlations (activity of caspase-3-like proteins vs Cu, Zn concentration; number of depolarized mitochondria vs Cu concentration) and positive correlations (number of necrotic cells vs Cu concentrations; activity of CAT vs Zn ) were noted. The defense of males against high metal content and its prooxidative effects is based mainly on GSH and CAT. In females the antioxidative reactions are species-specific and depend mainly on high peroxidase activity and on stress protein level. The increase in the number of apoptotic cells in the midgut gland of female spiders from the heavily polluted site suggests the defensive role of this process in maintaining the proper functioning of this organ.

Selenium modifies glutathione peroxidase activity and glutathione concentration in mice exposed to ozone-provoked oxidative stress.

The aim of this study was to show the direct effect of selenium on glutathione peroxidase (GSH-Px) activity and GSH/GSSG concentrations in 3- and 6-month-old mice. An ozone-oxygen mixture was used to provoke an oxygen stress. To measure the Se-effect mice were gavaged with sodium selenite. GSH-Px activity and total glutathione concentrations were determined in serum and in the postnuclear fraction of liver and lungs. Additionally glutathione concentrations were determined in whole blood. Both ozone and selenium, administered separately, reduced GSH-Px activity in lungs of 6-month-old animals, while in young mice an opposite effect of Se was observed. Ozone administered jointly with Se did not influence GSH-Px activity in 6-month-old mice, while in young, 3-month-old mice, a stimulatory effect in lungs was observed. There were no significant changes in GSH-Px activity in the liver of 6-month-old mice, but the stimulatory effect occurred in young mice treated with Se and Se & ozone jointly. In young mice, ozone (also ozone with Se) augmented glutathione concentrations. The response to ozone and selenium strictly depended on age and the antagonism between selenium and ozone was observed only in a few cases.