Alkaline DNase Activity Research Articles

Protective role of glutathione in oxidative stress caused by cadmium and copper

IntroductionCadmium is defined as one of the leading toxic industrial pollutants (Valko i sar., 2005). Although some products containing cadmium can be recycled, much of the pollution with this metal is the result of inadequate disposal and uncontrolled incineration of cadmium-containing waste (Jarup, 2003). Copper particles are released into the atmosphere from copper smelters and ore processing facilities, as well as from anthropogenic sources (use of pesticides, herbicides and fungicides). Oxidative stress occurs due to increased production of reactive oxygen species (Parkinson’s and Alzheimer’s disease) or reduced ability of cells to neutralize it through their internal antioxidants (eg mutation of the superoxide dismutase gene in amyotrophic lateral sclerosis).ObjectivesThe aim of this research was to examine the protective role of supplement, GSH, S-donor ligand, and in conditions of acute and chronic intoxication with sublethal doses of cadmium-II-chloride and copper II sulfate.MethodsAfter medial laparotomy albino rates Wistar soy, a 10% homogenate of brain tissue was made in an appropriate medium and an analysis of acid and alkaline DNase activity was performed (Kocić i sar., 2004; Kocić i sar., 1998).ResultsThis experiment demonstrated the beneficial role of GSH supplement that exhibit antioxidant character in preventing and reducing the adverse effects of acute and chronic cadmium and copper intoxication.ConclusionsAntioxidants prevent the formation of oxidative stress in the cell by reducing and stopping DNA damage and degradation, and thus represent potential scavengers of free radicals.DisclosureNo significant relationships.

Physiological strength of lipoic acid in copper intoxication

IntroductionThe average concentration of copper in the air, e.g. in the USA it ranges from 5–20 ng/m3, in soil from 5–70 mg/kg, and the intake of copper from food is 1.0–1.3 mg/days for adults (0.014–0.019 mg/kg/day) (Barceloux, 1999). The effect of lipoic acid is reflected in the intensification of ATP synthesis, participates in the assimilation of lactic acid, activates the enzyme cycle of tricarboxylic acid, stimulates the growth of lactic acid bacteria by replacing acetate (acetate transfer factor), stimulates CoA synthesis (fatty acid utilization), prevents liver damage by various toxins, normalizes aldolase and transferase levels.ObjectivesThe aim of this study is to show the useful role of a supplement, lipoic acid, as an antioxidant in the prevention of oxidative stress.MethodsAll procedures were performed after anesthesia of albino rats with ketal in accordance with the principles of sacrifice in laboratories. After medial laparotomy albino rates Wistar soy, a 10% homogenate of brain tissue was made in an appropriate medium and an analysis of acid and alkaline DNase activity was performed (Kocić i sar., 2004).ResultsDNases are thought to be the main executors of apoptosis, responsible for internucleosomal DNA fragmentation, which is the breakdown of chromosomal DNA into oligonucleosome-sized fragments. Administration of lipoic acid has been shown to protect against oxidative stress caused by copper.ConclusionsBased on the results of this research, it can be concluded that lipoic acid is a powerful and powerful antioxidant.DisclosureNo significant relationships.

Comparison of toxic effects of lead and copper and protective power of glutathione on oxidative stress parameters

IntroductionLead as an industrial pollutant can be detected at all stages of the working and living environment. Lead, based on its properties, solubility and mobility, accumulates in the soil, so that the average concentrations of oil in the soil are between 15 and 25 mg/kg (Radojević at all., 1999). Due to increased human activity, the amount of copper in the air, soil and water has increased. Glutathione (GSH) is an essential cofactor of many enzymes, such as: formaldehyde dehydrogenase, glyoxalase, prostaglandin endoperoxide isomerase, dehydrochlorinase and others. GSH is a biological redox in the metabolism of erythrocytes, it also plays a role in the transport of amino acids. Reactive forms of oxygen cause oxidative biomolecules (lipids, proteins, DNA) (Freidovich, 1999; Massaad i Klann, 2010).ObjectivesThe aim of this research was to examine the protective role of supplements GSH in conditions of chronic intoxication with sublethal doses of lead acetate and copper II sulfate.MethodsThe preparation of biomaterials for testing and making homogenates of brain tissue of albino rats of Wistar strain was performed and the activity of acid and alkaline DNase was measured spectrophotometrically (Kocić i sar., 1998).ResultsLead otherwise “as soft Lewis acid” has a pronounced affinity for interaction with “soft bases” such as S-atoms of the thiol group in antioxidants, natural biomolecules and supplements in this case in glutathione.ConclusionsIt can be said that GSH is a desirable supplement and antioxidant in the detoxification of reactive oxygen species in rats exposed to lead poisoning.DisclosureNo significant relationships.

Glutathione as a powerful antioxidant in oxidative stress in the brain tissue of rats caused by the pathophysiological action of copper

IntroductionDue to increased human activity, the amount of copper in air, soil and water has increased. Copper, at minimum concentrations, is essential for the normal functioning of the organism (cellular respiration, hemoglobin formation, growth and reproduction). At higher concentrations, copper is deposited in the liver, brain tissue, and bone marrow.ObjectivesTo investigate the protective role of the supplement, glutathione (GSH), the S-donor ligand, in conditions of chronic copper intoxication via the parameters of oxidative stress, ie. Alkaline and acidic DNase values in brain tissue in albino rats of Wistar strain.MethodsThe model system for testing the effects of copper exposure and the protective effect of GSH was a study on female albino rats of Wistar strain, stored in the vivarium of the Scientific Research Center for Biomedicine, Faculty of Medicine, Niš, Serbia. Endonucleases, alkaline and acidic DNase activities were determined spectrophotometrically from homogenates of brain tissue.ResultsCopper is believed to be a likely cause of oxidative damage to the DNA molecule, as manifested by increased alkaline and acidic DNase activity. The results of this study show that GSH is a potent chelator that binds copper and enables its elimination from the body.ConclusionsIn this experiment, the beneficial role of GSH supplements, which has an antioxidant character, in the prevention and reduction of the adverse effects of chronic copper intoxication was demonstrated. In this way, GSH acts as a powerful protector and antioxidant.DisclosureNo significant relationships.

Melatonin inhibits apoptosis and oxidative tissue damage in cisplatin-induced pulmonary toxicity in rats.

Cisplatin is one of the most frequently used chemotherapeutics, which is known to cause both tumor and normal lung tissue damage through the generation of free radicals and cells apoptosis/necrosis. Melatonin is a neurohormone that regulates numerous physiological processes in the body both through receptor pathways and by maintaining tissue redox homeostasis. The extent of rat lung damage induced by cisplatin and the effects of melatonin on this process was determined based on the pathohistological changes and biochemical disturbances in tissue lipid peroxidation, protein carbonyl modification and in the activity of xanthine oxidase (XO), caspase-3 and DNases. Histopathological analysis of rat lung tissue obtained from animals that received cisplatin found them to be edematous, with significant deterioration of alveolar epithelium. These morphological changes are accompanied by a significant increase in all studied oxidative stress-related parameters, as well as with the activity of apoptosis-related enzymes. A five-day treatment with melatonin completely prevented a cisplatin-induced increase in oxidative stress-related parameters and in the activity of XO, caspase-3 and alkaline DNase. Also, the histopathological changes observed during microscopic analysis were much less pronounced than in the group that received cisplatin only. These results can potentially be connected with the ability of melatonin to inhibit the activity of XO, caspase-3 and alkaline DNase and/or its ability to scavenge free radicals, thus preventing lung damage induced by cisplatin.

Depurinized milk downregulates rat thymus MyD88/Akt/p38 function, NF-\u03baB-mediated inflammation, caspase-1 activity but not the endonuclease pathway: in vitro/in vivo study

The aim of this study was the evaluation of 15 days dietary regimen of depurinized (DP) milk (obtained using our patented technological procedures) or 1.5% fat UHT milk instead of standard chow diet, on rat thymus and bone marrow MyD88/Akt/p38, NF-κB, caspase-1 and endonuclease pathways, in relation to peripheral blood cell composition. To determine whether the reduced mass of the thymus is a consequence of the direct effect of DP/UHT milk on apoptosis of thymocytes, in vitro Annexin-V-FITC/PI assay was performed. Significant decreases in the thymus wet weight, thymocyte MyD88, Akt-1/phospho-Akt-1 kinase, p38/phospho-p38, NF-κB, caspase-1 activity and CD4+/CD8+ antigen expression were obtained, especially in the DP milk group. The activity of thymocyte alkaline and acid DNase increased in the DP but not in the UHT milk group. The level of IL-6 significantly decreased in DP milk treated group, while the level of total TGF-β and IL-6 increased in UHT milk group. Significant differences in hematological parameters were obtained in commercial milk fed group. Observed results about prevention of experimental diabetes in DP pretreated groups may suggest that purine compounds, uric acid and other volatile toxic compounds of commercial milk may suppress oral tolerance, probably via IL-6 and TGF-β cytokine effects.

The Effects of Melatonin on Oxidative Stress Parameters and DNA Fragmentation in Testicular Tissue of Rats Exposed to Microwave Radiation.

Microwaves from mobile phones are one of the environmental toxicants that are capable of compromising male fertility by inducing oxidative stress and apoptosis in the testes. Melatonin is a lipophilic tryptophan indole amine and a potent antioxidant. The aim of the study was to evaluate the effect of melatonin treatment on oxidative stress parameters and DNA fragmentation in the testicular tissue of rats exposed to microwave radiation (4 h/day). Adult Wistar rats were divided in 4 groups: I--treated with saline; II--treated with melatonin; III--exposed to microwaves; IV--exposed to microwaves and treated with melatonin. The melatonin (2 mg/kg ip) was administered daily. The animals were sacrificed after 20, 40 and 60 days. Melatonin treatment prevented previously registered increases in malondialdehyde after only 20 days. Furthermore, it reversed the effects of microwave exposure on xanthine oxidase (after 40 days) and acid-DNase activity (after 20 days). However, neither protein carbonyl content nor catalase and alkaline Dnase activity were changed due to melatonin treatment. Melatonin exerts potent antioxidant effects in the testes of rats exposed to microwaves by decreasing the intensity of oxidative stress; it also reduces DNA fragmentation.

Enzymatic, antimicrobial and toxicity studies of the aqueous extract of Ananas comosus (pineapple) crown leaf

Ethnopharmacological relevanceVarious parts of the plant pineapple (Ananas comosus) are used in traditional medicine worldwide for treatment of a number of diseases and disorders. In folk medicine, pineapple leaf extract was used as an antimicrobial, vermicide, purgative, emmenagoogue, abortifacient, anti-oedema and anti-inflammatory agent. Compared to the fruit and stem extracts of pineapple, information about its leaf extract is limited. The potential of pineapple crown leaf extract as an ethno-medicine has been evaluated in terms of its enzymatic activities related to wound healing, antimicrobial property and toxicity. Materials and methodsMajor protein components of the extract were revealed by 2-D gel electrophoresis followed by MS/MS analysis. Zymography, DQ-gelatin assay were performed to demonstrate proteolytic, fibrinolytic, gelatinase and collagenase activities. DNase and RNase activities were revealed from agarose gel electrophoresis. Antimicrobial activity was evaluated spectrophotometrically from growth inhibition. Sprague-Dawley rat model was used to measure acute and sub-acute toxicity of the extract by analyzing blood markers. ResultThe extract contains several proteins that were clustered under native condition. Proteomic studies indicated presence of fruit bromelain as major protein constituent of the extract. It showed nonspecific protease activity, gelatinolytic, collagenase, fibrinolytic, acid and alkaline phosphatase, peroxidase, DNase and RNase activities along with considerable anti-microbial property. The leaf extract did not induce any toxicity in rats after oral administration of acute and sub-acute doses. ConclusionPineapple leaf extract is nontoxic, contains enzymes related to damage tissue repairing, wound healing and possibly prevents secondary infections from microbial organisms.

Melatonin protects rat thymus against oxidative stress caused by exposure to microwaves and modulates proliferation/apoptosis of thymocytes

The aim of the study was to evaluate the effect of melatonin on oxidative stress, DNA fragmentation, apoptsis and proliferation in thymus tissue of rats exposed to microwaves. Wistar rats were divided in four groups: I - treated with saline; II - treated with melatonin; III - microwaves exposed; IV - microwaves exposed and melatonin treated. Melatonin (2 mg/kg i.p.) was administered daily. Animals were sacrificed after 20, 40 and 60 days. A significant increase in malondialdehyde and carbonyl group content, as well as decrease in catalase and increase in xanthine oxidase activity were registered under microwave exposure. Melatonin prevented the increase in malondialdehyde and carbonyl group content, and reversed the effect on catalase and xanthine oxidase activity. Both, alkaline and acid DNase activity were increased due to microwave exposure. Furthermore, microwaves caused increase in apoptosis rate (detected using Annexin V-FITC/PI kit) and reduced proliferative capacity of thymocytes (induced by ConA). However, melatonin caused decrease in alkaline and acid DNase activity, decrease in apoptotic rate and increase in proliferation rate of thymocytes. Melatonin exerts protective effects on rat thymocytes by modulating processes of apoptosis and proliferation, and causes decrease in DNA fragmentation and oxidative stress intensity under exposure to microwaves.

Monitoring the effects of exposure to lead and cadmium in working and living environment through standard biochemical blood parameters and liver endonucleases activity

Heavy metals as pollutants in the working and living environment are a serious health and environmental problem because they are toxic, non-biodegradable, accumulate in living systems and have a long half-life in soil. Sources of lead contamination are combustion products in the chemical industry and metallurgy, industrial waste water, landfills, traffic etc. Lead enters into the body via the food chain and drinking water. In the body lead is deposited in the liver, kidneys, brain and mineral tissues. Excretion of lead causes damage to the epithelial cells of certain organs. High level exposure to cadmium is usually the result of environmental pollution by human activities. Exposure to cadmium can lead to acute and chronic tissue damage of various organs, including liver and kidneys in humans and in animals. In this paper we analyzed the effects of lead and cadmium exposure, in working and living environment, on the model system of experimental animals, particularly the activity of certain liver enzymes, acid and alkaline DNase, and standard biochemical blood parameters. The study showed that lead and cadmium significantly affect the protein content, red blood cells, hemoglobin and hematocrit, and the activity of liver enzymes. This harmful effect of this toxic metal can be reduced by the supplements.

Polymorphisms of tumor-necrosis factor-α−308 and lymphotoxin-α + 250: Possible modulation of susceptibility to apoptosis in chronic lymphocytic leukemia and non-Hodgkin lymphoma mononuclear cells

Tumor necrosis factor alpha (TNF-α) and lymphotoxin alpha (LT-α) have been shown to play an important role in the pathogenesis of limphoproliferative disease. Both cytokines regulate cell-survival and cell-death in leukemic cells. TNF-α and LT-α are highly produced in chronic lymphotic leukemia (CLL) and non-Hodgkin lymphoma (NHL) patients. Genetic polymorphism within regulatory regions of these cytokine genes can alter their expression levels. This study investigates an influence of TNF-α−308 and LT-α + 250 polymorphisms on the activity of alkaline DNase in mononuclear cells of both patient groups as a potent biochemical marker of DNA fragmentation in the terminal phase of apoptosis. Study was performed on mononuclear cells of CLL and NHL patients. SNP were obtained by PCR-RFLP method. The activity of alkaline DNase was measured by spectrophotometric method. The study provided evidence of the influence of TNFG/A genotype and A alleles in the susceptibility to NHL, since the association of LT-αG/G genotype with CLL was observed. High-producing TNF-α−308/LT-α + 250 heterozygous haplotype is associated with high NHL incidence. The investigated SNP influence the activity of alkaline DNase in CLL and NHL patients. The observed polymorphisms may modulate susceptibility of leukemic cells to apoptosis by way of DNase activity.

Sodium nitroprusside and peroxynitrite effect on hepatic DNases: an in vitro and in vivo study

BackgroundIt has been documented that nitric oxide (NO) donor sodium nitroprusside (SNP) and authentic peroxynitrite are capable of promoting apoptosis in a number of different cell types. Various endonucleases have been proposed as candidates responsible for the internucleosomal cleavage of the genomic DNA observed during apoptosis, but the main effect is attributed to the alkaline-DNases (Mg2+- and caspase-dependent) and acid-DNase. The aim of this study was to examine an in vivo and in vitro possibility for alkaline- and acid-DNases to be activated by SNP and peroxynitrite.ResultsThe effect on liver tissue alkaline and acid DNase activity together with the markers of tissue and plasma oxidative and nitrosative stress (lipid peroxidation, SH group content, carbonyl groups and nitrotyrosine formation) was investigated in plasma and liver tissue. The activity of liver alkaline DNase increased and that of acid DNase decreased after in vivo treatment with either SNP or peroxynitrite. A difference observed between the in vivo and in vitro effect of oxide donor (i.e., SNP) or peroxynitrite upon alkaline DNase activity existed, and it may be due to the existence of the "inducible" endonuclease. After a spectrophotometric scan analysis of purified DNA, it was documented that both SNP and peroxynitrite induce various DNA modifications (nitroguanine formation being the most important one) whereas DNA fragmentation was not significantly increased.ConclusionAlkaline DNase activation seems to be associated with the programmed destruction of the genome, leading to the fragmentation of damaged DNA sites. Thus, the elimination of damaged cells appears to be a likely factor in prevention against mutation and carcinogenesis.

In vivo reactivation of DNases in implanted human prostate tumors after administration of a vitamin C/K(3) combination.

Human prostate cancer cells (DU145) implanted into nude mice are deficient in DNase activity. After administration of a vitamin C/vitamin K(3) combination, both alkaline DNase (DNase I) and acid DNase (DNase II) activities were detected in cryosections with a histochemical lead nitrate technique. Alkaline DNase activity appeared 1 hr after vitamin administration, decreased slightly until 2 hr, and disappeared by 8 hr after treatment. Acid DNase activity appeared 2 hr after vitamin administration, reached its highest levels between 4 and 8 hr, and maintained its activity 24 hr after treatment. Methyl green staining indicated that DNase expression was accompanied by a decrease in DNA content of the tumor cells. Microscopic examination of 1-microm sections of the tumors indicated that DNase reactivation and the subsequent degradation of DNA induced multiple forms of tumor cell death, including apoptosis and necrosis. The primary form of vitamin-induced tumor cell death was autoschizis, which is characterized by membrane damage and the progressive loss of cytoplasm through a series of self-excisions. These self-excisions typically continue until the perikaryon consists of an apparently intact nucleus surrounded by a thin rim of cytoplasm that contains damaged organelles.

Evaluation of Serum Alkaline DNase Activity in Treatment Monitoring of Head and Neck Cancer Patients

Our previously published data on breast cancer suggest that serum alkaline DNase, a known circulating tumour marker, can be used for treatment monitoring of cancer patients. Serum alkaline DNase activities were analyzed in 215 untreated head and neck cancer patients. The enzyme activity ranged from 0.17 to 97.97 IKU/l in untreated cancer patients. Responders (n = 314) showed significantly elevated activity of alkaline DNase as compared to untreated cancer patients (p < 0.001). While non-responders (n = 168) showed comparable activity with untreated cancer patients. Serum alkaline DNase activities were significantly elevated in responders as compared to non-responders (p < 0.001). Our clinical studies during follow-up of patients indicated that the variations in serum alkaline DNase activities in individual patients correlate closely with response to therapy. Serum alkaline DNase also appeared to be useful in predicting treatment response in the long-term follow-up of patients. Serum alkaline DNase was systematically examined as a possible indicator for recurrence in patients under complete remission. In conclusion, serum alkaline DNase may be useful as a treatment monitoring in patients with head and neck malignancies.

Herpesvirus alkaline deoxyribonuclease; a possible candidate as a novel target for anti-herpesvirus therapy.

Herpesvirus alkaline deoxyribonucrease (DNase) is coded in the genome of all herpesvirus species determined total sequence and is conserved in structure. In order to determine whether the enzyme could be a target for a novel antiherpesvirus therapy, the anti-herpes simplex virus type 1 (HSV-1) activity of antisense oligonucleotide for HSV-1 alkaline DNase was studied. Six antisense phosphorothioate oligonucleotides, targeted to an internal AUG start codon, were designed and evaluated. One of the oligonucleotides, UL12-4, inhibited wild type and thymidine kinase-deficient HSV-1 replication to 21.5 and 19.5% at 40 microM, respectively. The quantity of alkaline DNase mRNA and DNase activity in HSV-1-infected Vero cells was reduced to one eighth and 66.9% of control, respectively, by treatment with 40 microM of UL12-4, but no effect was observed on the quantity of HSV-1 glycoprotein H mRNA (gamma2 gene) or on the replication of Vero cells. These results indicate that UL12-4 inhibits HSV-1 replication by decreasing the amount of alkaline DNase mRNA. The herpesvirus alkaline DNase could be a novel target for anti-herpesvirus drug.

Influences of diet and surgical trauma on serum alkaline DNase activity levels

Changes in serum alkaline DNase activities might predict the therapeutic response in various malignant diseases. A decrease in serum alkaline DNase activity within days from the onset of therapy has been related to tumour necrosis and may be a possible sign of clinical response to effective treatment. To study if changes in serum alkaline DNase activity could be induced by non-tumour related tissue destruction, sera were collected on several occasions perioperatively in 18 patients undergoing surgery for benign gynaecological disease. Thirty apparently healthy women served as the control group. A significant decrease (P less than 0.001) in serum alkaline DNase activity was observed after an overnight fast in both groups of women. In contrast to the control women, the operated patients showed a significant decrease (P less than 0.001) in serum alkaline DNase activity throughout the operative period and 1 week postoperatively. We conclude that serum alkaline DNase activity is influenced by dietary factors as well as surgical trauma. These factors may limit the clinical usefulness of SADA in patients with cancer.

Initial experiences with serum alkaline DNase activity in monitoring the effects of therapy for carcinoma of the uterine cervix

The objective was to evaluate if variations in serum alkaline DNase activity (SADA) can predict the effects of therapy in women with early stages of primary cervical carcinoma. 29 out of 33 patients had no evidence of disease after therapy. Only 5 out of the 29 women showed increased SADA levels after therapy compared with the pretreatment SADA value. Of the 4 women with evidence of disease after therapy, 3 had unchanged or decreased SADA levels. We conclude that serum alkaline DNase activity seems to have little to offer in predicting the effects of treatment in stage I and stage II cervical carcinoma.

Variations in serum alkaline DNase activity have preditive value in the monitoring of clinical response to cancer therapy : B. Dewez, A. Iaraoglou, M. Lana, H. Taper, M. Robertroid. UCL 7369-1200 Bruxelles

Variations in serum alkaline DNase activity have preditive value in the monitoring of clinical response to cancer therapy : B. Dewez, A. Iaraoglou, M. Lana, H. Taper, M. Robertroid. UCL 7369-1200 Bruxelles

Predictive Value of Serum Alkaline Dnase Activity Variations in Treatment of Head and Neck Cancer

The aim of this study was to evaluate the variation in serum alkaline DNase activity (SADA) as a means of therapeutic monitoring in patients with head and neck cancer. Blood samples from 40 patients were collected before, during, and some weeks up to months after therapy. A decrease in SADA during treatment was usually associated with a primary clinical response, while no decrease indicated non-response to therapy. In patients with complete tumor regression the initial decrease of SADA was usually followed by an increase exceeding the initial level. A similar increase was not observed in patients with tumor progression.

Variations in serum alkaline DNase activity: a new means to assess early detection of relapse in patients treated for acute nonlymphoblastic leukemia

Our previously published clinical results on various malignancies indicated that the variations in serum alkaline DNase activity (SADA) could be a sensitive test for therapeutic monitoring of human malignancies. In the present study, the clinical efficacy of SADA detecting relapse in 32 acute nonlymphoblastic leukemia (ANLL) patients in remission was tested. The observation period ranged from 3 to 17 months. A simple and rapid biochemical technique based on spectrophotometric measurements was used to assay SADA. Of the 32 patients, 17 remained in remission and had less than a 15% variation in SADA levels. They had no clinical symptoms of recurrence at any time. In the remaining 15 patients, after a period of stability, a progressive decrease in SADA, with variations of more than 15%, was observed without any treatment. At that time, no abnormalities of clinical parameters were detected in these patients. A recurrence of disease as evidenced by routine examinations was found relatively late after the first decrease in SADA in all 15 patients (range 1.5 to 5.5 months). These results suggest that periodic measurements of SADA during the posttherapeutic course can be used as a means to assess early detection of an eventual recurrence.