The Impact of Element–Element Interactions on Antioxidant Enzymatic Activity in the Blood of White Stork (Ciconia ciconia) Chicks

The aim of this work was to determine interrelationships among macroelements Na, K, Ca, Mg, and Fe, microelements Zn, Cu, Mn, and Co, and toxic heavy metals Pb and Cd in the blood of white stork Ciconia ciconia, during postnatal development, in different Polish environments, and their impact on the activity of antioxidant enzymes. We considered the content of thiobarbituric acid-reactive substances (TBARSs), i.e., malondialdehyde (MDA), and activity of superoxide dismutase (SOD), catalase (CAT), ceruloplasmine (CP), glutathione peroxidase (GPx), and glutathione reductase (GR). Blood samples were collected from storks developing at Odra meadows (Kłopot; southwestern Poland). They were compared with blood of chicks from several suburban sites located 20 km away from Zielona Góra (0.1 million inhabitants; southwestern Poland) and near Głogów, where a copper smelter is situated. We also conducted research in the Pomeranian region (Cecenowo; northern Poland). We collected blood samples via venipuncture of the brachial vein of chicks in 2005–2007. They were retrieved from the nest and placed in individual ventilated cotton sacks. The blood was collected using a 5-ml syringe washed with ethylenediaminetetraacetic acid (EDTA). We found significant interactions between macro- and microelements and enzymatic activity and TBARS products. We noticed the predominance of Cd and Pb participation in element–enzyme interactions. Simultaneously, we found interrelationships between cadmium and Na, K, Ca, Mg, and Fe and the activity of antioxidant enzymes SOD, CAT, CP, GR, and TBARS products in the blood of white stork chicks. In the case of lead these relationships were not numerous and they were significant for Ca, Mg, Cu, Mn, and Co. Correlations with enzymes were significant for Pb-CAT and Pb-TBARS. We noted that activities of most enzymes (SOD, CAT, CP, GR) and TBARS products are determined by their interactions with physiological elements Na, Ca, Mg, Fe, and Zn and toxic heavy metals. White stork chicks ranged in age from 17 to 59 days. Concentrations of elements in the blood were age related. Among enzymes, only SOD, CAT, and GPx were age related. Young storks differed in the case of element concentration (except for Ca, Zn, and Cd) and enzymatic activity. We found that significant element–element interaction/enzyme activity predominated in the case of physiological elements and toxic metals, which we explain by the intensive and prevailing access of toxic metals in redox reactions. This causes changes in the priority of these metals, reflected by their influence on the enzymatic activity of antioxidant enzymes. The content of Cd and Pb in blood of young storks from different regions tends to affect the lipid peroxidation process negatively. However, in many cases we observed an increase in enzymatic activity with an increase in heavy metals. This indicates the changes in oxidative stress intensity in chicks in response to environmental differentiation. The increase in lipoperoxidation modifies antioxidant enzyme activity and causes changes in SOD, CAT, CP, GPx, and GR activity in chicks from various regions, principally increases in enzyme activity in chicks from polluted environments and suburbs. We suggest that the source of heavy metals in chicks’ blood might be used as a biological test system of adaptation to oxidative stress. We also report that a high level of heavy metals is accompanied by increased lipid peroxidation. Thus young storks are probably significantly susceptible to environmental conditions. They demonstrated initiation of lipoperoxidation and oxidative modification of proteins that coincide with chemical elements, as a possible antioxidant defense system.     

Behavioral characterization of GLT1 (+/-) mice as a model of mild glutamatergic hyperfunction

GLT1 is one of the major transporters responsible for maintenance of glutamate homeostasis in the brain. In the present study, glutamate transporter 1-deficient GLT1 homozygous (-/-) and heterozygous (+/-) mice were investigated with the intention that they may provide a model of hyperglutamatergic state resulting in various behavioral alterations. The GLT1 (-/-) mice had lower body and brain weight, mild neuronal loss in CA1 hippocampal region as well as focal gliosis and severe focal neuronal paucity in layer II of the neocortex. The short life-span of GLT1 (-/-) precluded us from systematic behavioral studies in these mice. In contrast, GLT1 (+/-) mice exhibiting a 59% decrease in GLT1 immunoreactivity in their brain tissue, showed no apparent morphological brain abnormalities, and their life-span was not markedly different from controls. Behaviorally, GLT1 (+/-) presented moderate behavioral alterations compared to their wildtype littermates, such as: mild sensorimotor impairment, hyperlocomotion (at 3 month of age only), lower anxiety (at 6 months), better learning of cue-based fear conditioning but worse context-based fear conditioning. Our results suggest that GLT1 (+/-) mice may serve as a potentially useful model to study neurodegenerative disease conditions with mild hyperglutamatergic activity.     

Role of fosB in behaviours related to morphine reward and spatial memory

The immediate early genes (IEGs) have been suggested to be implicated in mechanisms of addiction, as well as in learning and memory processes. fosB, which belongs to IEG, has been reported to have pleiotropic impact on response to psychoactive drugs, as well as motivational and stress-related behaviours. In the present study, we used mice with constitutive knock-out of fosB in order to study fosB role in mouse phenotype. We studied rewarding properties of morphine (10 mg/kg i.p.) in conditioned place preference (CPP) paradigm. Additionally, we studied fosB role in spatial memory and spatial working memory using elevated plus maze model of spatial learning (EPMSL) and delayed non-match to place task (DNMTP). In further studies, locomotor, depressive-like and anxiety-like behaviours were measured. Rewarding effects of morphine in fosB −/− mice were abolished whereas spatial learning was impaired. On the other hand, we found no significant differences in locomotor activity, depression-like and anxiety-like behaviours. In summary, our results indicate that mice lacking fosB are less sensitive to rewarding properties of morphine and display spatial memory impairment and suggest involvement of fosB and its proteins in motivational aspects of reinforcers as well as in learning and memory processes.     

Gene expression profile as a prognostic factor in high-grade gliomas

Some clinical factors have been useful in predicting prognosis in high-grade gliomas, however, unexpected differences in survival time have generated attempts to search for more precise parameters. It is clear that tumour behaviour depends mostly on gene alterations. Known single gene alterations failed to accurately define survival time, however, recently, the gene profiling based on microarray technology has raised hopes. Our aim was to assess whether the genetic predictor exceeds clinical parameters in the prognosis of malignant gliomas. We performed gene expression analysis of 28 gliomas (3 grade II, 10 grade III and 15 grade IV, according to WHO classification), and 5 control, normal brain samples, using Clontech oligonucleotide arrays with 3,757 known genes. The signal-to-noise statistics was used to separate classes, and the leave-one-out method was used to assess the smallest number of genes make it clear with a minimal cross-validation error. All gliomas, or only high-grade tumours, were clearly separated from the normal brain samples using 7 or 9 most differentially expressed genes. Hierarchical clustering failed, but the fuzzy c-means method was useful in high-grade gliomas to find a gene prediction model, which, with clinical factors, was assessed in survival analysis. Univariate analysis demonstrated that age, WHO grade (IV vs. III), radiation dose (> or = 50 Gy vs. 42 Gy), postoperative KPS score (100 points vs. others), neurological deficit as the first sign of the disease vs. others, and gene expression profile were significant predictors of survival. In multivariate analysis, the gene expression profile remained the only independent predictor (p = 0.007). Thus, our conclusion is that gene expression pattern predicts outcome in high-grade gliomas independently of other factors.     

Posturography and computerized gait analysis in the Computer Dyno Graphy system as non-invasive methods for evaluation of normal pressure hydrocephalus progression

Disturbances of gait and balance are important clinical problems in patients with normal pressure hydrocephalus. A considerable degree of locomotor disability and increased risk of falls are present in many cases. Accordingly, attempts to develop effective evaluation methods of gait and balance disturbances are made. Posturography and the Computer Dyno Graphy (CDG) system appear to be useful in diagnosis and evaluation of treatment in patients with normal pressure hydrocephalus. Both are non-invasive and repeatable methods that enable accurate evaluation of analyzed parameters at time intervals.     

Polymorphism in the P-glycoprotein drug transporter MDR1 gene in renal transplant patients treated with cyclosporin A in a Polish population

P-glycoprotein (P-gp), the product of MDR1 gene, is a protein which mediates transmembrane transport of a great number of xenobiotics including cyclosporin A used as an immunosuppressive drug in patients with allogenic kidney grafts. The P-gp activity and expression is dependent on the MDR1 gene polymorphism in position C3435T of exon 26. In this study, C3435T polymorphism was analyzed in 116 patients with allogenic kidney graft treated with cyclosporin Aand 144 randomly selected healthy individuals. The prevalence of MDR1 gene genotypes 3435CC, 3435CT, 3435TT were also compared in patients after allogenic kidney graft with both acute and chronic graft rejection (48 patients with acute and 76 with chronic graft rejection) and control groups (respectively 139 and 112). The results of the study demonstrated that the allelic frequency and MDR1 genotype distribution were similar in all evaluated groups. It was revealed that MDR1 gene polymorphism was not a predisposing factor for terminal kidney failure leading to renal transplantation. Moreover, evaluation of C3435T polymorphism of MDR1 gene will probably not be useful for characterization of groups of patients at increased risk of acute and chronic kidney graft rejection.     

Dye-assisted lymph vessels sparing laparoscopic varicocelectomy

OBJECTIVE: Hydrocele, the main complication of laparoscopic varicocelectomy, is thought to result from a disruption of gonadal lymphatics. The aim of this study was to evaluate the effectiveness of patent blue V dyeing to identify and preserve lymphatic vessels and to assess whether the lymphatic sparing technique avoids postoperative hydrocele in adolescent boys undergoing a laparoscopic procedure. MATERIALS AND METHODS: Fifty-two (52) boys affected by varicocele Grade III (range, 12-16 years) underwent a left-sided laparoscopic varicocelectomy. Twenty-six (26) boys were randomly assigned to a lymphatic nonsparing (LNS) group, and the others to a lymphatic sparing (LS) group. Before surgery in the LS group, 2 mL of patent blue V was injected under the tunica dartos on the left side. RESULTS: All varicocelectomies were performed laparoscopically. Lymphatic vessels were identified in 23 (88.5%) boys of the LS group. In the remaining three (11.5%), the lymphatics could not be identified clearly. No adverse local or generalized reactions were noted. At a mean follow-up of 14 months, no recurrent varicocele or testicular volume reduction were detected. Hydrocele developed in 4 LNS patients and 1 was operated on. No patient from the LS group developed a hydrocele. CONCLUSIONS: Staining gonadal lymph vessels with patent blue V is an effective and simple method of visualization of the lymphatic drainage from the testis. Blue-stained lymph vessels could be readily distinguished and preserved during a laparoscopic varicocelectomy, which results in a decrease of hydrocele development. To validate an efficacy of vital staining of lymphatic vessels in avoiding hydrocele formation, a larger series and longer follow-up are necessary.     

Short-term effects of angiotensin II receptor blockade in patients with primary glomerulonephritis: pilot study.

OBJECTIVE: There is evidence that angiotensin II, the main effector of renin-angiotensin system, plays a crucial role in the pathogenesis of chronic renal injury. Angiotensin II type 1 (AT-1) receptor blockers reduce renin-angiotensin system activity by blocking the receptor, the activation of which is responsible for the majority of deleterious angiotensin II effects. The aim of the present study was to investigate renal and metabolic effects of specific AT-1 receptor blocker-losartan in patients with primary glomerulonephritis. DESIGN: Pilot clinical study. SETTING: Department of Nephrology, Transplantology and Internal Medicine, Medical University of Gdańsk, Poland. PARTICIPANTS: Fifteen patients aged 43.3 +/- 11.3 years with primary glomerulonephritis confirmed by renal biopsy were studied. INTERVENTION: Creatinine clearance, urinary excretion of protein and uric acid, urinary activity of N-acetyl-beta-D-glucosaminidase, and serum concentrations of lipids, protein, and uric acid were evaluated before and after 3 months of losartan (Cozaar; Merck Sharp & Dohme, Harlow, Essex, United Kingdom) treatment at a dose of 25 mg daily. RESULTS: We found a significant reduction of urinary excretion of protein (P <.008; average, 32%). Results also revealed a decrease of serum uric acid level (P <.01), probably as a consequence of elevated uric acid urinary excretion (P <.09). No significant changes in creatinine clearance, N-acetyl-beta-D-glucosaminidase activity, protein, or lipid levels were observed. CONCLUSION: We concluded that losartan treatment at a small dose of 25 mg daily produces antiproteinuric effect without adverse effects, notably with no decrease of glomerular filtration, and simultaneously induces beneficial changes in purine metabolism.