Arsenic: toxicity, oxidative stress and human disease.

Arsenic (As) is a toxic metalloid element that is present in air, water and soil. Inorganic arsenic tends to be more toxic than organic arsenic. Examples of methylated organic arsenicals include monomethylarsonic acid [MMA(V)] and dimethylarsinic acid [DMA(V)]. Reactive oxygen species (ROS)-mediated oxidative damage is a common denominator in arsenic pathogenesis. In addition, arsenic induces morphological changes in the integrity of mitochondria. Cascade mechanisms of free radical formation derived from the superoxide radical, combined with glutathione-depleting agents, increase the sensitivity of cells to arsenic toxicity. When both humans and animals are exposed to arsenic, they experience an increased formation of ROS/RNS, including peroxyl radicals (ROO•), the superoxide radical, singlet oxygen, hydroxyl radical (OH•) via the Fenton reaction, hydrogen peroxide, the dimethylarsenic radical, the dimethylarsenic peroxyl radical and/or oxidant-induced DNA damage. Arsenic induces the formation of oxidized lipids which in turn generate several bioactive molecules (ROS, peroxides and isoprostanes), of which aldehydes [malondialdehyde (MDA) and 4-hydroxy-nonenal (HNE)] are the major end products. This review discusses aspects of chronic and acute exposures of arsenic in the etiology of cancer, cardiovascular disease (hypertension and atherosclerosis), neurological disorders, gastrointestinal disturbances, liver disease and renal disease, reproductive health effects, dermal changes and other health disorders. The role of antioxidant defence systems against arsenic toxicity is also discussed. Consideration is given to the role of vitamin C (ascorbic acid), vitamin E (α-tocopherol), curcumin, glutathione and antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase in their protective roles against arsenic-induced oxidative stress.

Antioxidant properties of carotenoids: QSAR prediction of their redox potentials.

There is a great need to predict the antioxidant properties of molecules such as carotenoids. These compounds are of great interest due to their contribution to various important biological and industrial processes, including toxicity and fate. In our study, redox potentials were compiled from several literature sources. Redox potential values ranged from 537.2 mV for zeaxanthin up to 691.5 mV for beta-carotene; they correspond to the formation of cation radicals, using the standard calomel electrode (SCE). The redox potential values were measured using conventional electrochemical techniques, cyclic voltammetry and Osteryoung square-wave voltammetry. A quantitative structure-activity relationship (QSAR) was developed to model and consequently to predict the values of redox potential. The predicted values of redox potential for four external carotenoids, namely beta-carotene, zeaxanthin, cantaxanthin and astaxanthin, are presented and discussed. They indicate the dependence of redox potential on structure, donor and acceptor groups and polarisability.

Free radicals, metals and antioxidants in oxidative stress-induced cancer.

Oxygen-free radicals, more generally known as reactive oxygen species (ROS) along with reactive nitrogen species (RNS) are well recognised for playing a dual role as both deleterious and beneficial species. The "two-faced" character of ROS is substantiated by growing body of evidence that ROS within cells act as secondary messengers in intracellular signalling cascades, which induce and maintain the oncogenic phenotype of cancer cells, however, ROS can also induce cellular senescence and apoptosis and can therefore function as anti-tumourigenic species. The cumulative production of ROS/RNS through either endogenous or exogenous insults is termed oxidative stress and is common for many types of cancer cell that are linked with altered redox regulation of cellular signalling pathways. Oxidative stress induces a cellular redox imbalance which has been found to be present in various cancer cells compared with normal cells; the redox imbalance thus may be related to oncogenic stimulation. DNA mutation is a critical step in carcinogenesis and elevated levels of oxidative DNA lesions (8-OH-G) have been noted in various tumours, strongly implicating such damage in the etiology of cancer. It appears that the DNA damage is predominantly linked with the initiation process. This review examines the evidence for involvement of the oxidative stress in the carcinogenesis process. Attention is focused on structural, chemical and biochemical aspects of free radicals, the endogenous and exogenous sources of their generation, the metal (iron, copper, chromium, cobalt, vanadium, cadmium, arsenic, nickel)-mediated formation of free radicals (e.g. Fenton chemistry), the DNA damage (both mitochondrial and nuclear), the damage to lipids and proteins by free radicals, the phenomenon of oxidative stress, cancer and the redox environment of a cell, the mechanisms of carcinogenesis and the role of signalling cascades by ROS; in particular, ROS activation of AP-1 (activator protein) and NF-kappaB (nuclear factor kappa B) signal transduction pathways, which in turn lead to the transcription of genes involved in cell growth regulatory pathways. The role of enzymatic (superoxide dismutase (Cu, Zn-SOD, Mn-SOD), catalase, glutathione peroxidase) and non-enzymatic antioxidants (Vitamin C, Vitamin E, carotenoids, thiol antioxidants (glutathione, thioredoxin and lipoic acid), flavonoids, selenium and others) in the process of carcinogenesis as well as the antioxidant interactions with various regulatory factors, including Ref-1, NF-kappaB, AP-1 are also reviewed.

Oxygen free radical generating mechanisms in the colon: do the semiquinones of vitamin K play a role in the aetiology of colon cancer?

It is proposed that bile acids (deoxycholic acid), the K vitamins, iron(II) complexes and oxygen interact to induce an oncogenic effect in the colon by the generation of free radicals. In the relatively low oxidising/reducing conditions of the colonic lumen the K vitamins exist in the reduced form; however, if absorbed into the mucosa they have the capacity to be chemically oxidised and to enter into a redox cycle yielding oxygen radicals. The semiquinone radical of K(1) (phylloquinone) has been stabilised in bile acid mixed micelles and investigated by electron paramagnetic resonance spectroscopy and quantum chemical calculations. The estimated half-life of the radical was about 30 min which confirms a remarkably high stability in aqueous micellar solution. A model is presented in which the reduced K vitamins may initiate superoxide radical, O2(-*) generation leading to Fe(II) mediated Fenton reactions in the stem colon cells.

Metals, toxicity and oxidative stress.

Metal-induced toxicity and carcinogenicity, with an emphasis on the generation and role of reactive oxygen and nitrogen species, is reviewed. Metal-mediated formation of free radicals causes various modifications to DNA bases, enhanced lipid peroxidation, and altered calcium and sulfhydryl homeostasis. Lipid peroxides, formed by the attack of radicals on polyunsaturated fatty acid residues of phospholipids, can further react with redox metals finally producing mutagenic and carcinogenic malondialdehyde, 4-hydroxynonenal and other exocyclic DNA adducts (etheno and/or propano adducts). Whilst iron (Fe), copper (Cu), chromium (Cr), vanadium (V) and cobalt (Co) undergo redox-cycling reactions, for a second group of metals, mercury (Hg), cadmium (Cd) and nickel (Ni), the primary route for their toxicity is depletion of glutathione and bonding to sulfhydryl groups of proteins. Arsenic (As) is thought to bind directly to critical thiols, however, other mechanisms, involving formation of hydrogen peroxide under physiological conditions, have been proposed. The unifying factor in determining toxicity and carcinogenicity for all these metals is the generation of reactive oxygen and nitrogen species. Common mechanisms involving the Fenton reaction, generation of the superoxide radical and the hydroxyl radical appear to be involved for iron, copper, chromium, vanadium and cobalt primarily associated with mitochondria, microsomes and peroxisomes. However, a recent discovery that the upper limit of "free pools" of copper is far less than a single atom per cell casts serious doubt on the in vivo role of copper in Fenton-like generation of free radicals. Nitric oxide (NO) seems to be involved in arsenite-induced DNA damage and pyrimidine excision inhibition. Various studies have confirmed that metals activate signalling pathways and the carcinogenic effect of metals has been related to activation of mainly redox-sensitive transcription factors, involving NF-kappaB, AP-1 and p53. Antioxidants (both enzymatic and non-enzymatic) provide protection against deleterious metal-mediated free radical attacks. Vitamin E and melatonin can prevent the majority of metal-mediated (iron, copper, cadmium) damage both in vitro systems and in metal-loaded animals. Toxicity studies involving chromium have shown that the protective effect of vitamin E against lipid peroxidation may be associated rather with the level of non-enzymatic antioxidants than the activity of enzymatic antioxidants. However, a very recent epidemiological study has shown that a daily intake of vitamin E of more than 400 IU increases the risk of death and should be avoided. While previous studies have proposed a deleterious pro-oxidant effect of vitamin C (ascorbate) in the presence of iron (or copper), recent results have shown that even in the presence of redox-active iron (or copper) and hydrogen peroxide, ascorbate acts as an antioxidant that prevents lipid peroxidation and does not promote protein oxidation in humans in vitro. Experimental results have also shown a link between vanadium and oxidative stress in the etiology of diabetes. The impact of zinc (Zn) on the immune system, the ability of zinc to act as an antioxidant in order to reduce oxidative stress and the neuroprotective and neurodegenerative role of zinc (and copper) in the etiology of Alzheimer's disease is also discussed. This review summarizes recent findings in the metal-induced formation of free radicals and the role of oxidative stress in the carcinogenicity and toxicity of metals.

A technique for the fast sampling of biological tissues for electron paramagnetic resonance spectroscopy.

A simple technique for the fast sampling of biological tissues for electron paramagnetic resonance (EPR) spectroscopy is described. By using tube-like stainless steel cutter, a cylindrical piece of tissue is dissected free; while holding the sample and cutter on a silicone rubber cutting base, the sample is transferred directly into the EPR test tube by pushing the tube into the instrument. The main features of the technique are its simplicity, speed, elimination of the necessity to manipulate frozen tissue (e.g., breaking, chopping, etc.), precise information on the sample volume, the ability to adjust sample size according to need, and avoidance of the use any foreign compounds (such as spin traps). The technique has been successfully applied to the EPR detection of free radicals in rabbit spinal cord exposed to ischemia. It can be easily modified for manipulation with samples in an inert atmosphere.

Analysis of the radial and longitudinal effect in a double TE(104) and a single TE(102) rectangular cavity.

The response of the cavity to the rotation of a point-like sample in the horizontal (y-z) plane passing through the center of the Bruker double TE(104) and single TE(102) rectangular cavities in concentric circles of radii rho = 0, 1, 2, 3, 4, and 5 mm from the cavity center (radial effect) has been analyzed. The experimentally observed dependencies of the EPR signal intensity, I(pp), showed the following: (i) for rho = 0 mm (a sample position in the cavity center), I(pp) is independent of the angle of rotation; (ii) for rho = 1, 2, and 3 mm, the I(pp) dependence progressively changes from circular to oval; (iii) when the radius is further increased to rho = 4 and 5 mm, the I(pp) dependence changes dramatically, giving a figure eight shape. These experimental observations are in very good agreement with the theoretical calculations, in which the response is modeled using modified Cassinian curves, K(rho, phi). Similar trends were observed for any position of the horizontal (y-z) plane at which the sample is situated along the vertical x axis of the cavity; however, the amplitude of the signal decreases with increase in the absolute value of the x coordinate, ||x ||. The variation in the signal amplitude along the cavity x axis (longitudinal effect) can be calculated theoretically using a modified sine-squared curve, G(x). In general, the response of the cavity to a point-like sample situated at any position, P(rho, phi, x), can be represented as a product of the mentioned Cassinian curve, K(rho, phi), and sine-squared curve, G(x), giving for the signal intensity I(pp)(rho, phi, x) approximately K(rho, phi)G(x). The response to a large cylindrical sample which is concentrically situated on the cavity x axis can then be obtained by integrating the above product, K(rho, phi)G(x), over the sample volume. The nonlinear radial effect may give rise to a serious source of systematic error in quantitative EPR spectroscopy and shows that accurate and precise positioning of the sample in the microwave cavity is essential.

[Rubber band ligation of hemorrhoids in ambulatory care]. / Elastická ligatúra hemoroidálnych uzlov v ambulantnej praxi.

OBJECTIVE: The aim of the study was to evaluate the outcome of elastic ligation in internal haemorrhoids in our out-patient setting. METHODS: The authors performed elastic ligation of haemorrhoids in 77 patients, and of these 71 (92%) procedures were accomplished in out-patients. The procedure was indicated in second and third-degree symptomatic haemorrhoids and was carried out without local anaesthesia with the aid a Seward Medical applicator. Typically, one haemorrhoidal node was treated per one session. All patients were clinically examined two weeks after completion of treatment. RESULTS: The success rate of the procedure was 91%. The results were analyzed by a questionnaire. The most common unpleasant symptoms were pain and tenesms. Bleeding occurred in 6 patients. There was no serious life-threatening complication. CONCLUSION: Based on our observation we consider band ligation as a reliable therapy for symptomatic internal haemorrhoids in out-patients. We suggest reminding patients of possible of complications, unpleasant symptoms sensations restricting their daily activity on the first days after the procedure.

[Gastrointestinal surgical complications after kidney transplantation]. / Gastrointestinálne chirurgické komplikácie po transplantácii oblicky.

BACKGROUND AND OBJECTIVE: Clinical spectrum of gastrointestinal (GI) complication in renal transplant recipients ranges from nonspecific signs of abdominal discomfort to life-threatening surgical emergencies. The manifestation of such complications is modified by antirejection therapy with resulting risk of diagnostic and therapeutic error. The present study analyses the incidence of serious GI complications after renal transplantation and their treatment. METHODS: Retrospective analysis in a university transplant center in years 1988-2000. There was 239 renal transplants performed in 234 patients (age 42 +/- 14). Male to female ratio was 140.99. Polycystic kidney disease patients comprised 9% (n = 22). RESULTS: Five serious GI complications resulting in death were recorded (2x upper GI bleed, 1x acute pancreatitis, 1x bowel obstruction in sclerotizing peritonitis, 1x diffuse purulent peritonitis of unknown origin). There was no case of diverticulitis. The average time onset of lethal GI complication was 2.8 +/- 1.4 years after renal transplantation. There was no statistically significant relation to underlying diagnosis of renal failure, warm and cold ischemia time, and time elapsed after transplantation. CONCLUSION: GI complications have increased incidence in patients after renal transplantation. The incidence of complications has no relationship to underlying diagnosis of renal failure, warm and cold ischemia time, and time elapsed after transplantation. Patients involved in transplant program should be carefully followed and thoroughly examined with any new GI symptomatology. Further studies are necessary to evaluate potential of pre-transplant screening programs.

Analysis of the movement of line-like samples of variable length along the X-axis of a double TE104 and a single TE102 rectangular cavity

Movement of line-like samples with lengths from 5 to 50 mm along the x-axis of the double TE104 rectangular cavity has been analyzed. The observed dependencies of the EPR signal intensity versus sample position showed: (i) a sharp maximum for sample lengths from 5 to 20 mm; (ii) a plateau, over which the EPR signal intensity remained constant within experimental errors of 0.26-1.07%, for lengths from 30 to 40 mm; and (iii) a "sloping plateau," which could be approximated by the linear function (correlation, r = 0.98) for sample length 50 mm. Theoretical values of the experimentally observed dependencies of the intensity versus sample position were calculated using the modified sine-squared function and the correlation between observed and theoretically predicted dependencies is very good. The experimental dependence of the EPR signal intensity versus the sample length for samples situated at the same point in the cavity was nonlinear with a maximum for the 40-mm sample. The dependence of the EPR signal intensity upon the movement of a large cylindrical sample (o.d. 4 mm and length 100 mm) along the x-axis of the cavity was similar to that found for the 50-mm sample. However, an additional oscillating signal superimposed on the sloping plateau was observed. The presence of a large sample fixed in the complementary cavity of the double TE104 cavity caused an additional deformation of the signal intensity for a 30-mm sample which was moving in the first cavity. The primary effect was that the plateau was replaced by a region in which the intensity increased linearly with sample position, r = 0.99. Each of the above phenomena may be a source of significant errors in quantitative EPR spectroscopy. Cylindrical samples to be compared should be of identical length and internal diameter. Accurate and precise positioning of each sample in the microwave cavity is essential. Copyright 1997 Academic Press. Copyright 1997Academic Press

Free radicals in rabbit spinal cord ischemia: electron spin resonance spectroscopy and correlation with SOD activity.

1. In nonanesthetized rabbits temporal occlusion of the abdominal aorta was used to induce oxidative stress in the lower part of the body including distal segments of the spinal cord. 2. Spinal cord samples were taken from the animals exposed to 25-min aortic occlusion (AO) or to occlusion followed by 1- or 2-hr reperfusion (AO/R1 or AO/R2, respectively) or from sham-operated animals (C). The presence of free radicals (FR) in the spinal cord samples frozen in liquid N2 was assessed by ESR spectroscopy without spin trapping. Moreover, superoxide dismutase (SOD) activity and conjugated diene (CD) levels were measured in the samples. 3. In the AO group FR were detected in the spinal cord regions close to the occlusion (lower thoracic and distal segments) along with a decrease in SOD activity. The calculated g value (g = 2.0291) indicated that the paramagnetic signal recorded might be attributed to superoxide radicals. FR were absent in the AO/R1 group. Concurrently, the SOD activity revealed a significant tendency to return to the control level. FR appeared again in the AO/R2 group, mostly in the upper and middle lumbar regions, along with a decrease in SOD activity. No sample from the C group revealed FR. A significant increase in CD levels was observed in the thoracolumbar region only in the AO/R2 group. The temporary absence of FR in the AO/R1 group suggests activation of defense antioxidant mechanisms (e.g., specific enzymatic systems such as SOD), which might have been exhausted later. 4. Changes in SOD activity similar to those observed in the thoracolumbar region, though less noticeable, occurred in the obviously noncompromised tissue (upper cervical region). This points to a kind of generalized response of the animal to aortic occlusion. 5. Direct ESR spectroscopy revealed the presence of FR as well as their time course in the spinal cord during the early phase of ischemia/reperfusion injury and the inverse relationship between FR and SOD activity.

1H NMR ganglioside ceramide resonance region on the differential diagnosis of low and high malignancy of brain gliomas.

1. The high-resolution 1H NMR (MRS) spectra of human brain tumor homogenates revealed a broad resonance at 5.3-5.4 ppm in glioblastoma multiforme (N = 16) and brain metastases (N = 3). The broad resonance was identified as ceramide, a sphingosine-fatty acid combination portion of ganglioside, indicating an elevated abundance of monounsaturated fatty acids. GLC analysis of gangliosides in the highly malignant glioblastoma multiforme revealed that the elevated monounsaturated fatty acid is oleic acid (C18:1). The resonance at 5.3-5.4 ppm region was not detectable in normal human brain (N = 2), in meningiomas (N = 2), or in low-grade astrocytomas (N = 12). In normal human brain the abundance of monounsaturated fatty acid is minimal. 2. This investigation was made possible because the method of producing homogenate resulted in (i) no loss of lipids during the process and (ii) a well-homogenised sample, with (iii) no loss in chemical integrity. 3. The properties of tumor gangliosides include antigenic specificity and immunosuppressive activity and the ceramide, a sphingosine-fatty acid combination, noticeably influences the ganglioside immunosuppressive activity. 4. The observation of 1H NMR ceramide resonance in high-malignant brain tumors emphasizes the dramatic role of aberant gangliosides and ceramide precursors on the grade of malignancy and invasiveness. 5. Further insight into the specific nature of the ceramide portion of gangliosides in grading the malignancy of brain tumors should be investigated further.