Cell cycle inhibitory effects of leaf extract from Curcuma vamana M. Sabu & Mangaly on mitotically synchronous cultures of Physarum polycephalum Schw.

Leaf extracts of C. vamana, endemic to Kerala state in India, were found to inhibit cell cycle progression in synchronous cultures of P. polycephalum in a concentration and phase-specific manner. Crude alkaloid extract (CAE) elicited maximum cell cycle delays in comparison to soxhletted chloroform, acetone and aqueous extracts. Total alkaloid content of CAE was found to be 64.9 mg/g. CAE showed lowest DPPH radical scavenging activity. Other extracts with higher free radical scavenging activity exhibited lesser cell cycle inhibiting potential. Upto 21% decrease in nuclear DNA was observed in CAE treated samples. However, genotoxicity as evidenced by comet assay was not observed. The extracts were also found to be non-toxic to human RBCs at the highest concentration tested (750 µg/mL). CAE treatment completely suppressed a 63 kDa polypeptide with a concomitant, but weak induction of a 60 kDa polypeptide suggesting that these may be cell cycle related. CAE was found to possess potent antiproliferative activity against PBLs. The study clearly demonstrates the cell cycle inhibitory activity of C. vamana leaf extracts, with CAE being the most potent of them.

Protective role of vitamins C and E in diclorvos-induced oxidative stress in human erythrocytes in vitro

Organophosphate (OP) pesticides such as dichlorvos (DDVP) intoxication has been shown to produce oxidative stress due to the generation of free radicals, which alter the antioxidant defense system in erythrocytes. In this study, the effects of DDVP (1, 10, 100 µM) or DDVP + vitamin C (VC; 10 µM) or vitamin E (VE; 30 µM), on the levels of malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities in human erythrocytes were examined in vitro. There were no statistical differences between all groups for 1 µM concentration of DDVP. Treatment with DDVP alone produced an increase in the level of MDA and decreased activities of antioxidant enzymes (P < 0.05). Groups treated with vitamins and DDVP showed protective effects of vitamins against DDVP-induced changes in antioxidant enzyme activity and lipid peroxidation (LPO) (10 µM). At 100 µM concentration of DDVP vitamins had no effect on DDVP-induced toxicity. The results show that administration of DDVP resulted in the induction of erythrocyte LPO and alterations in antioxidant enzyme activities, suggesting that reactive oxygen species (ROS) may be involved in the toxic effects of DDVP. Also the data show that the plasma level of VC and VE may ameliorate OP-induced oxidative stress by decreasing LPO in erythrocytes at certain doses of OP pesicides.

In vitro antioxidant activity of banana (Musa spp. ABB cv. Pisang Awak).

The methanolic extract of Musa ABB cv Pisang Awak was investigated for the polyphenolic contents and antioxidant activity. The total phenol and flavonoid contents of the fruit extract were found to be 120 mg gallic acid equivalents (GAE) and 440 mg quercetin equivalents (QE)/100 g of sample dry weight, respectively. The antioxidant activity of the Pisang Awak methanol extract (PAME) (20-500 µg/ml) was determined using 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity, reducing capacity, 2-2’-azinobis-3-ethylbenzothiozoline-6-sulfonic acid (ABTS) radical cation decolourization and hydroxyl radical scavenging capacity (OH·). The EC50 values of DPPH, ABTS and OH· activities of the PAME and butylated hydroxy toluene (BHT) were found to be 65 and 9 µg/ml, 29 and 6 µg/ml, 36 and 42 µg/ml respectively. The reducing capacity increased with increasing concentration (31.5-1000 mg/ml) of the fruit extract and the activity was comparable with the standard BHT. The high performance thin layer chromatography (HPTLC) analysis of the extract revealed the presence of polyphenols. The strong and positive correlations were obtained between total phenol/flavonoid contents (R2 = 0.693-1.0) and free radical scavenging ability was attributed to the polyphenols as the major antioxidants.

Carbohydrate content and antioxidative potential of the seed of three edible indica rice (Oryza sativa L.) cultivars.

Rice (Oryza sativa L.) grains or seeds are known to lose much of their nutrient and antioxidant contents, following polishing. The current study was undertaken to evaluate and compare the carbohydrate content and antioxidant parameters in the unpolished and polished seeds of three edible indica rice cultivars, namely Swarna (SW), the most popular indica rice cultivar in India and aromatic or scented cultivars Gobindobhog (GB) and Pusa Basmati (PB). While both the sucrose and starch content was the maximum in PB seeds (both unpolished and polished), the amylose content was the highest in SW polished seeds. SW polished seeds were superior as compared to GB and PB cultivars in terms of total antioxidant capacity, DPPH radical scavenging and Fe(II) chelation potential, as well as the highest lipoxygenase (LOX) inhibition or H2O2 scavenging potential, probably due to the maximum accumulation of total phenolics and flavonoids, the two important antioxidants. The reducing power ability was, however, identical in both SW and GB polished seeds. The PB polished seeds were more potent in superoxide and hydroxyl scavenging, whereas GB in nitric oxide (NO) scavenging. The common observation noted after polishing of seeds was the reduction in the level of carbohydrates and antioxidant potential, though the extent of reduction varied in the three cultivars. The only exception was GB, where there was no alteration in NO scavenging potential even after polishing. Our study showed the better performance of SW polished seeds with respect to higher amylose content and majority of the tested parameters governing antioxidant capacity and radical scavenging potential, thus highlighting the greater dietary significance of SW over the other two cultivars.

Toxic effects of mercury, lead and gadolinium on vascular reactivity

Heavy metals have been used in a wide variety of human activities that have significantly increased both professional and environmental exposure. Unfortunately, disasters have highlighted the toxic effects of metals on different organs and systems. Over the last 50 years, the adverse effects of chronic lead, mercury and gadolinium exposure have been underscored. Mercury and lead induce hypertension in humans and animals, affecting endothelial function in addition to their other effects. Increased cardiovascular risk after exposure to metals has been reported, but the underlying mechanisms, mainly for short periods of time and at low concentrations, have not been well explored. The presence of other metals such as gadolinium has raised concerns about contrast-induced nephropathy and, interestingly, despite this negative action, gadolinium has not been defined as a toxic agent. The main actions of these metals, demonstrated in animal and human studies, are an increase of free radical production and oxidative stress and stimulation of angiotensin I-converting enzyme activity, among others. Increased vascular reactivity, highlighted in the present review, resulting from these actions might be an important mechanism underlying increased cardiovascular risk. Finally, the results described in this review suggest that mercury, lead and gadolinium, even at low doses or concentrations, affect vascular reactivity. Acting via the endothelium, by continuous exposure followed by their absorption, they can increase the production of free radicals and of angiotensin II, representing a hazard for cardiovascular function. In addition, the actual reference values, considered to pose no risk, need to be reduced.

Efecto de la vitamina C, sobre la actividad de glutation peroxidasa y la formación de ateromas, en conejos expuestos a dieta hiperlipidémica / Effects vitamin C, on glutathione peroxidase activity and atheroma formation, in rabbits exposed to a hyperlipidaemic diet

El estrés oxidativo juega un papel muy importante en la aterosclerosis; de hecho existe evidencias que indican que los antioxidantes son moléculas capaces de retardar y/o revertir el proceso aterosclerótico. El objetivo del presente estudio fue comparar el efecto de la vitamina C (Vit C), sobre la actividad de la Glutation peroxidasa (GPx) y la formación de ateromas en conejos. Se estudiaron 36 conejos divididos en 3 grupos: Grupo 1 (Control): conejarina, Grupo 2: huevo y conejarina, Grupo 3: huevo, conejarina y Vit C (100mg/diarios). el período experimental duró 12 semanas. Se determinó perfil lipídico por métodos enzimáticos y la actividad de GPx por cinética en 0 y 12va semana. Los conejos fueron sacrificados y se les realizó estudio histológico de su aorta. Los resultados revelaron un incremento en la actividad de la GPx en los grupos 2 y 3 con respecto al control en la 12va semana de experimentación (p<0,05). Hubo inhibición de lesiones ateroscleróticas en los conejos del grupo 3. En conclusión en condiciones de hiperlipidemia con o sin suplementación de Vit C, existe incremento en la actividad de GPx. Por otra parte, la Vit C disminuye y evita la progresión de ateromas.

Oxidative stress plays an important role in artherosclerosis; so antioxidants are molecules have been used to slow down or inihibit atherosclerosis. The objetive of the presents study was to compare the effect of Vitamin C (Vit C), on serum Glutathione peroxidase activity (GPx) and on the formation of aortic lesions in rabbits. 36 rabbits were studied: Group 1: "conejarina" (commercial rrabit food); Group 2: egg and conejarina, Group 3: egg, conejarina and Vit C (100mg/day). The experimental lasted 12 weeks. Lipid profile was done by enzymatic methods and GPx by kinetic method in weeks 0 and 12. Histological study of rabbit's aorta was done. GPx activity in groups 2 and 3, increased compared with controls, from weeks 12 of experimentation (o<0,05). There was inihition of aortic lesions in groups 3. In conclusion, under hyperlipidemic conditions, with or without Vit C supplementation, activity of GPx there is increase. Vit C reduces and prevents the progression of atheromas.

Evaluation of free radical scavenging activity of Butea monosperma Lam.

In the present study, ethyl acetate, butanol and aqueous fractions derived from total methanol extract of Butea monosperma flowers were evaluated for radical scavenging activities using different in vitro models like reducing power assay, scavenging of 2,2 diphenyl-1-picrylhydrazyl (DPPH) radical, nitric oxide radical, superoxide anion radical, hydroxyl radical and inhibition of erythrocyte hemolysis using 2, 2' azo-bis (amidinopropane) dihydrochloride (AAPH). Methanol extract along with its ethyl acetate and butanol fractions showed potent free radical scavenging activity, whereas aqueous fraction was found to be devoid of any radical scavenging properties. The observed activity could be due to the higher phenolic content in the extracts (16.1, 25.29, and 17.74% w/w in methanol extract, ethyl acetate and butanol fractions respectively). HPTLC fingerprint profile of the ethyl acetate and butanol fractions were developed which would serve as reference standard for quality control of the extracts.

Peroxynitrite: a potent oxidizing and nitrating agent.

Nitric oxide (NO*) reacts with superoxide (O2-*) forming peroxynitrite (PXN) (ONOO-), a strong oxidant which reacts with several biomolecules leading to enormous implications in biological process, holds enormous implications for the understanding of free radicals. The ONOO- formation in vivo has significant implications in free radical biology. It exerts a defensive role in large number of pathophysiological reactions and also acts as signaling molecule in activation of several protooncogenes. It decomposes rapidly to an intermediate and reacts with several biomolecules. Evidence for PXN formation in vivo has been obtained immunohistochemically through detection of a characteristic reaction product with protein tyrosine residues and 3-nitrotyrosine. This "biomarker" of PXN formation has now been identified in various pathologies such as Lou Gehrig's disease, Parkinson's disease, cancer, atherosclerosis as well as in biological aging. 3-nitrotyrosine formation has been documented in various tissues, e.g. even in non-diseased embryonic heart during normal development. Therefore, there is a great opportunity in the postgenomic period to understand the interplay of these molecular interactions with biological events such as apoptosis, gene regulation etc. This review deals with biological significance of peroxynitrite, its precursors, reactions with large range of biomolecules, including aminoacids, proteins, lipids, nucleic acids, antioxidants as well as cytotoxic aspects.

Antioxidant potential of C-phycocyanin isolated from cyanobacterial species Lyngbya, Phormidium and Spirulina spp.

The antioxidant activity of C-Phycocyanin (C-PC) isolated from three cyanobacterial species Lyngbya (marine), Phormidium (marine) and Spirulina (fresh water) was studied in vitro. The results demonstrate that C-PCs from Lyngbya, Phormidium and Spirulina spp. are able to scavenge peroxyl radicals (determined by crocin bleaching assay) with relative rate constant ratio of 3.13, 1.89 and 1.8, respectively. C-PCs also scavenge hydroxyl radicals (determined by deoxyribose degradation assay) with second order rate constant values of 7.87 x 10(10), 9.58 x 10(10) and 6.42 x 10(10), respectively. Interestingly, Lyngbya C-PC is found to be an effective inhibitor of peroxyl radicals (IC50 6.63 microM), as compared to Spirulina (IC50 12.15 microM) and Phormidium C-PC (IC50 12.74 microM) and is close to uric acid (IC50 2.15 microM). Further, the studies suggest that the covalently-linked tetrapyrrole chromophore phycocyanobilin is involved in the radical scavenging activity of C-PC. The electron spin resonance (ESR) spectra of C-PCs indicate the presence of free radical active sites, which may play an important role in its radical scavenging property. This is the first report on the ESR activity of native C-PCs without perturbations that can cause radical formation.

Development of semi-empirical model for radiochemical reactions

A mathematical model, which represents the radio-chemical reactions in water, was developed to study the effect of the radio-chemical products on cell kiling. The five differential equations were solved using dose rate equation and cell survival as a function of dose was computed. The known chemical rate constants were taken from the literature and unknown constants were determined by curve fitting to an experimental data. Sensitivity studies were performed by varying the rate constants and showed that the yield of H-radical had little effect whereas the change in concentration of OH-radical and direct interaction resulted in significant change on cell survival. The sensitivity studies showed good agreement with the observed effects. In conclusion, we developed a mathematical model that could be used as a means for the estimation of radiation damage.

Radicales libres y antioxidantes en la prevención de enfermedades: I mecanismos de generación de radicales libres / Free radicals and antioxidants in disease prevention: I free radical generating mechanisms

Los radicales libres y las especies derivadas del oxígeno (ERO) son continuamente generadas en el organismo como resultado de accidentes de la química, o durante el curso de diversos procesos metabolicos. Las ERO han sido implicadas como iniciadoras de reacciones que conducen a un daño oxidativo, y por lo tanto se considera que dichas especies subyacen al desarrollo de diversas condiciones patológicas. Sin embargo, recientemente, estas especies han sido reconocidas también como importantes mediadores de una serie de eventos de naturaleza fisiológica. El hecho de que las ERO conduzcan a un daño celular, o que sus acciones se limiten a servir como señales biológicas, dependerá en parte de su naturaleza, de los mecanismos y sitios de su generación, y de los sustratos biológicos sobre los que actúan. No obstante, el mayor determinante de que las ERO tengan o no carácter deletéreo esta dado por el balance entre la velocidad a la cual estas especies son generadas y aquella a la cual las mismas son removidas por los mecanismos antioxidantes. En el presente trabajo, el primero de una serie de tres, los autores cubren las principales definiciones esenciales en relación a radicales libres y especies derivadas del oxígeno. Adicionalmente, revisan el conocimiento actual sobre la química y la biología de la generación de estas especies, sus acciones sobre sustratos celulares, y discuten en forma crítica el doble papel que las ERO juegan, ya sea, como mediadores de eventos biológicos esenciales, o como iniciadores de procesos oxidativos conducentes a fenómenos cito-deletéreos

Phosphate radical induced oxidation of pyrimidine bases in aqueous solution.

The photooxidation of pyrimidine bases viz., uracil and cytosine in presence of peroxydiphosphate (PDP) in aqueous solution at natural pH (approximately 7.5) has been carried out in a quantum yield reactor using a high-pressure mercury lamp. The rates of oxidation and quantum yields of pyrimidine oxidation have been found to increase with increase in [PDP] while they are independent of [pyrimidine] and light intensity. On the basis of these experimental results, product analysis and existence of isosbestic points a probable mechanism is suggested in which peroxydiphosphate ion on photolysis gives phosphate radical anions which initiates the reaction by adding to C(5) or C(6) of pyrimidine base leading to the formation of pyrimidine radical via radical cation or hydrolysis. This further reacts with PDP and gives the final products 5,6-dihydroxy pyrimidine and isobarbituric acid.

Biochemistry of free radicals: from electrons to tissues

Free radicals are chemical species with an unpaired electron in the outer valence orbitals. The unpaired electron makes them paramagnetic (physics) and relatively reactive (chemistry). The free radiclas that are normal metabolites in aerobic biological systems have varied reactivities, ranging from the high reactivity of hydroxyl radical (t(1/2) = 10(-9) s)) to the low reactivity of melanins (t (1/2) = days)). The univalent reduction of oxygen that takes place in mammalian organs produces superoxide radicals at a rate of about 2 per cent of the total oxygen uptake. The primary production of superoxide radicals sustains a free radical chain reaction involving a series of reactive oxygen species (hydrogen peroxide, hydroxyl and peroxyl radical and singlet oxygen). Nitric oxide is almos tunreactive as free radical except for its termination reaction with superoxide radical to yield the strong oxidant peroxynitrite. Nitric oxide also reacts with ubiquinol in a redox reaction, with cytochrome oxidase competitively with oxygen, and oxymyoglobin and oxyhemoglobin displacing oxygen. Septic shock and endotoxemia produce muscle dysfunction and oxidative stress due to increased steady state concentrations of reative oxygen and nitrogen species.

Photochemistry and photobiology of actinic erythema: defensive and reparative cutaneous mechanisms

Sunlight is part of our everyday life and most people accept it as beneficial to our health. With the advance of our knowledge in cutaneous photochemistry, photobiology and photomedicine over the past four decades, the terrestrial solar radiation has become a concern of dermatologists and is considered to be a major damaging environmental factor for our skin. Most photobiological effects (e.g., sunburn, suntanning, local and systemic immunosuppression, photoaging or dermatoheliosis, skin cancer and precancer, etc.) are attributed to ultraviolet radiation (UVR) and more particularly to UVB radiation (290-320 nm). UVA radiation (320-400 nm) also plays an important role in the induction of erythema by the photosensitized generation of reactive oxygen species (singlet oxygen ((1)O2))superoxide (O2-) and hydroxyl radicals ((OH) that damage DNA and cellular membranes, and promote carcinogenesis and the changes associated with photoaging. Therefore, research efforts have been directed at a better photochemical and photobiological understanding of the so-called sunburn reaction, actinic or solar erythema. To survive the insults of actinic damage, the skin appears to have different intrinsic defensive mechanisms, among which antioxidants (enzymatic and non-enzymatic systems) play a pivotal role. In this paper, we will review the basic aspects of the action of UVR on the skin: a) photochemical reactions resulting from photon absorption by endogenous chromophores; b) the lipid peroxidation phenomenon, and c) intrinsic defensive cutaneous mechanisms (antioxidant systems). The last section will cover the inflammatory response including mediator release after cutaneous UVR exposure and adhesion molecule expression.

Bioquimica de los radicales libres / Biochemical of the free radicals

El concepto de que el oxigeno o sus productos puedan ser toxicos, para el medico que con frecuencia se enfrenta a enfermedades isquemico-hipoxicas del corazon, cerebro, etc., parece poco creible. Sin embargo, se han ampliado en forma considerable las evidencias que demuestran que los productos a partir del oxigeno puedan jugar un rol patogenico en multiples enfermedades

Los radicales libres: beneficios y problemas: III. Beneficios de los radicales libres del oxígeno / Free radicals: problems and benefits: III. Oxygen free radical benefits

Entre las respuestas inespecíficas de los organismos para la destrucción de los agentes patógenos se han identificado diversos procesos enzimáticos, unos responsables de la hidrólisis de macromoléculas y otros que permiten la producción de radicales libres, y por ende, una secuencia de procesos oxidativos que favorecen la actividad de las células fagocíticas. En este trabajo se analizan algunos mecanismos de síntesis de los radicales libres, sus efectos benéficos y algunos mecanismos de regulación. En particular, se revisa la participación fisiológica del radical libre, óxido nítrico, en los sistemas cardiovascular, nervioso e inmune.

Radicais livres e seu papel em cardiologia: atualizaçäo / Free radicals and its paper in cardiology: up-dating

É feita uma ampla revisão da atomística e dos aspectos químicos relacionados aos radicais livres. Discute-se ainda a influência dos radicais livres nos vários aspectos da Cardiologia e o papel potencial do uso de antioxidantes como recurso terapêutico.

Photoaffinity labeling and photoaffinity crosslinking of enzymes

Photoaffinity labeling is a special type of chemical modification, where the label is activated by the action of light. This article presents the general principles and limitations of this technique, its application to the study of Micrococcus luteus ATPase and the use of photoaffinity crosslinking to probe the structure of this enzyme