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.

Effects of melatonin on DNA damage induced by cyclophosphamide in rats

The antioxidant and free radical scavenger properties of melatonin have been well described in the literature. In this study, our objective was to determine the protective effect of the pineal gland hormone against the DNA damage induced by cyclophosphamide (CP), an anti-tumor agent that is widely applied in clinical practice. DNA damage was induced in rats by a single intraperitoneal injection of CP (20 or 50 mg/kg). Animals received melatonin during the dark period for 15 days (1 mg/kg in the drinking water). Rat bone marrow cells were used for the determination of chromosomal aberrations and of formamidopyrimidine DNA glycosylase enzyme (Fpg)-sensitive sites by the comet technique and ofXpf mRNA expression by qRT-PCR. The number (mean ± SE) of chromosomal aberrations in pinealectomized (PINX) animals treated with melatonin and CP (2.50 ± 0.50/100 cells) was lower than that obtained for PINX animals injected with CP (12 ± 1.8/100 cells), thus showing a reduction of 85.8% in the number of chromosomal aberrations. This melatonin-mediated protection was also observed when oxidative lesions were analyzed by the Fpg-sensitive assay, both 24 and 48 h after CP administration. The expression of Xpf mRNA, which is involved in the DNA nucleotide excision repair machinery, was up-regulated by melatonin. The results indicate that melatonin is able to protect bone marrow cells by completely blocking CP-induced chromosome aberrations. Therefore, melatonin administration could be an alternative and effective treatment during chemotherapy.

Antioxidant effect of 4-nerolidylcatechol and α-tocopherol in erythrocyte ghost membranes and phospholipid bilayers

4-Nerolidylcatechol (4-NC) is found in Pothomorphe umbellataroot extracts and is reported to have a topical protective effect against UVB radiation-induced skin damage, toxicity in melanoma cell lines, and antimalarial activity. We report a comparative study of the antioxidant activity of 4-NC and α-tocopherol against lipid peroxidation initiated by two free radical-generating systems: 2,2′-azobis(2-aminopropane) hydrochloride (AAPH) and FeSO4/H2O2, in red blood cell ghost membranes and in egg phosphatidylcholine (PC) vesicles. Lipid peroxidation was monitored by membrane fluidity changes assessed by electron paramagnetic resonance spectroscopy of a spin-labeled lipid and by the formation of thiobarbituric acid-reactive substances. When lipoperoxidation was initiated by the hydroxyl radical in erythrocyte ghost membranes, both 4-NC and α-tocopherol acted in a very efficient manner. However, lower activities were observed when lipoperoxidation was initiated by the peroxyl radical; and, in this case, the protective effect of α-tocopherol was lower than that of 4-NC. In egg PC vesicles, malondialdehyde formation indicated that 4-NC was effective against lipoperoxidation initiated by both AAPH and FeSO4/H2O2, whereas α-tocopherol was less efficient in protecting against lipoperoxidation by AAPH, and behaved as a pro-oxidant for FeSO4/H2O2. The DPPH (2,2-diphenyl-1-picrylhydrazyl) free-radical assay indicated that two free radicals were scavenged per 4-NC molecule, and one free radical was scavenged per α-tocopherol molecule. These data provide new insights into the antioxidant capacity of 4-NC, which may have therapeutic applications for formulations designed to protect the skin from sunlight irradiation.

Oxidative stress and cellular immunity in patients with recurrent aphthous ulcers

Recurrent aphthous ulcer (RAU) is an inflammatory condition of the oral mucosa characterized by painful, well-circumscribed, single or multiple round or ovoid ulcerations. The exact etiologic factor(s) of these ulcerations are not yet understood. The objective of this study was to evaluate inflammatory processes and free radical metabolism of 25 patients with RAUs compared to 25 healthy controls. The levels of malondialdehyde (MDA) and glutathione (GSH) were determined by high-performance liquid chromatography. Tumor necrosis factor-alpha (TNF-α), interleukin-2 (IL-2), IL-10, and IL-12 were determined by ELISA. Nitric oxide (NO), myeloperoxidase (MPO), total antioxidant status (TAS), and total oxidant status (TOS) levels were measured spectroscopically in serum. The levels of MDA, GSH, TNF-α, IL-2, IL-12, MPO, and TOS, and oxidative stress index (OSI) were higher, and the levels of NO, IL-10, and TAS were lower in patients with RAU than in controls. Statistical analysis showed that GSH, TNF-α, IL-2, IL-10, and OSI differed significantly in patients with RAU compared to controls. These parameters have important roles in oxidant/antioxidant defense.

Antioxidant capacity of anthocyanins from acerola genotypes

Anthocyanins from 12 acerola genotypes cultivated at the Active Germplasm Bank at Federal Rural University of Pernambuco were isolated for antioxidant potential evaluation. The antioxidant activity and radical scavenging capacity of the anthocyanin isolates were measured according to the β-carotene bleaching method and 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay, respectively. The antioxidant activity varied from 25.58 to 47.04% at 0.2 mg.mL-1, and it was measured using the β-carotene bleaching method. The free radical scavenging capacity increased according to the increase in concentration and reaction time by the DPPH assay. At 16.7 μg.mL-1 concentration and after 5 minutes and 2 hours reaction time, the percentage of scavenged radicals varied from 36.97 to 63.92% and 73.27 to 94.54%, respectively. Therefore, the antioxidant capacity of acerola anthocyanins varied amongst acerola genotypes and methods used. The anthocyanins present in this fruit may supply substantial dietary source of antioxidant which may promote health and produce disease prevention effects.

Chemical characterization of leaves of organically grown carrot Dacus carota L.) in various stages of development for use as food

In Brazil, street markets and vegetable distributors discard vegetable leaves and stems, including those of carrot (Dacus carota L.). Seeking to reduce the waste of vegetable parts, this study characterized chemically the leaves of organically grown carrot in three stages of development to determine the best time for their removal and consumption as food. The leaves were dehydrated in an oven at 70 °C for 43 hours and analyzed for chemical composition, antioxidant activity, chlorophyll content, fatty acid composition, and also calcium (Ca), sodium (Na), potassium (K), magnesium (Mg), manganese (Mn), iron (Fe), zinc (Zn), and copper (Cu) contents. The analyses indicated 100 days of development as the ideal stage for the removal and consumption of carrot leaves with good antioxidant activity requiring only 63.78 ± 0.5 mg.L-1 methanol leaf extract to inhibit 50% of the concentration of the free radical DPPH (2,2-diphenyl-1picrilidrazil), and total protein and alpha-linolenic acid (18:3 n-3/LNA) contents of 18.23% ± 2.8 and 876.55 ± 20.62 mg.100 g-1 of dry matter, respectively.

Evaluation of antioxidant activity of agro-industrial waste of acerola (Malpighia emarginata D.C.) fruit extracts

This study was carried out to evaluate the antioxidant capacity of the agro-industrial waste from acerola. Hydroacetone, hydroethanolic, and hydromethanolic extracts were obtained using the sequential extraction process, and they were screened for their free radical DPPH• (1,1-diphenyl-2-picrilhidrazil) and ABTS•+ (2,2'-azino-bis-(3-etilbenzotiazolin 6-sulfonic acid) scavenging activity and their effect on the linoleic acid peroxidation by the ferric thiocyanate method. Soybean oil with the addition of the extracts (200 ppm) was submitted to Schaal oven test (60 °C, 28 days), in which the samples were analyzed for peroxide value and conjugated dienes. Hydroethanolic and hydromethanolic extracts exhibited good DPPH scavenging activity (low value of EC50 and TEC50 and high value of AE), good ABTS scavenging capacity (1445.1 and 1145.5 µMol TEAC.g-1, respectively), and high percentage inhibition of peroxidation of linoleic acid (96.12 and 91.84%, respectively) and showed the ability to retard the formation of peroxides and conjugated dienes.

Phenolic compounds and antioxidant activity in fermented rice (Oryza sativa) bran

This study investigated the content of total phenolic compounds and antioxidant activity in fermented rice bran in order to evaluate the effect of solid state fermentation on these properties. The process was performed with the fungus Rhizopus oryzae CTT 1217 in tray reactors at 30 °C for 120 hours. Samples of fermented rice bran were collected every 24 hours. Antioxidant property was evaluated by the diphenyl-1-picrylhydrazyl radical scavenging method and through the inhibition of enzymatic oxidation and lipid peroxidation of olive oil. The methanol extract of the biomass obtained at 96 hours of fermentation inactivated 50% of free radical in 15 minutes. The same extract reduced the peroxide value in the olive oil by 57% after 30 days of storage. The aqueous extract of the biomass obtained at 120 hours was the most efficient inhibitor of the darkening reaction catalyzed by peroxidase.

Phenols, flavonoids and antioxidant activity of aqueous and methanolic extracts of propolis (Apis mellifera L.) from Algarve, South Portugal

Propolis is a resinous substance collected by honeybees to seal honeycomb, which has been used in folk medicine due to its antimicrobial and antioxidant properties. In the present study, water and methanol were used to extract phenols and flavonoids from propolis collected in thirteen different areas in the Algarve region during the winter and spring. The ABTS•+, DPPH•, and O2•- scavenging capacity, and metal chelating activity were also evaluated in the propolis samples. Methanol was more effective than water in extracting total phenols (2.93-8.76 mg/mL) (0.93-2.81 mg/mL). Flavones and flavonols were also better extracted with methanol (1.28-2.76 mg/mL) than with water (0.031-0.019 mg/mL). The free radical scavenging activity, ABTS (IC50=0.006-0.036 mg/mL), DPPH (IC50=0.007-0.069 mg/mL) and superoxide (IC50=0.001-0.053 mg/mL), of the samples was also higher in methanolic extracts. The capacity for chelating metal ions was higher in aqueous extracts (41.11-82.35%) than in the methanolic ones (4.33-29.68%). Propolis from three locations of Algarve region were richer in phenols and had better capacity for scavenging free ABTS and DPPH radicals than the remaining samples. These places are part of a specific zone of Algarve known as Barrocal.

Antioxidant activities of kombucha prepared from three different substrates and changes in content of probiotics during storage

Kombucha is a health-promoting fermented beverage worldwide. The present study compared the free-radical scavenging abilities and total reducing power (TRP) of kombucha prepared from low-cost green tea (LGTK), black tea (BTK), and tea powder (TPK). LGTK had the highest scavenging abilities against 2,2-diphenyl-1-picrylhydrazyl (DPPH), superoxide anion and hydroxyl radicals, while BTK showed the highest TRP. Changes in content of probiotics in LGTK were investigated during storage as well. The number of acetic acid bacteria decreased moderately up to 10 days of storage. The number of lactic acid bacteria (LAB) decreased significantly, and their survival rate was only 0.98% at the 8th day of storage.

Physicochemical properties and antioxidant activity of Tunisian date palm (Phoenix dactylifera L.) oil as affected by different extraction methods

The chemical analysis of flesh and seed of date palm fruit (Kentichi) was evaluated. Carbohydrates were the predominant component in all studied date cultivars (~78.69-83.46 g/100g dry matter), followed by moisture content (~9.23-11.17%), along with moderate amount of fat (~0.56-7.10 g/100g dry matter), protein (~2.16-2.80 g/100g dry matter), and ash (~1.18-1.64 g/100 g dry matter). Some antioxidants (Ascorbic acid, total phenolic, total flavonoid, chlorophyll and carotenoids) were found in different values in both date fruit and seed. The physicochemical properties and antioxidant activity of both flesh and seed oil which was extracted using Hexane, Soxhlet and Modified Bligh - Dyer extraction methods were determined. The experimental results showed that temperature, different solvents and extraction time had significant effect on the yield of the date palm oil and physicochemical properties. Date Flesh oil showed an important free radical scavenging activity towards 1-1-diphenyl-2-picrylhydrazyl (DPPH) free radical.

Influence of agro-ecological production areas on antioxidant activity, reducing sugar content, and selected phytonutrients of orange-fleshed sweet potato cultivars

Sweet potato is an important staple, and it is mainly known for its contribution of β-carotene in human diet. The effects of cultivar and habitat on this pigment and other nutritional characteristics of the crop still require investigation. In this study, three locally bred cultivars of sweet potato, two of which are orange-fleshed, were grown in three different agro-ecological areas to determine soluble sugar content, β-carotene, and total antioxidants of roots. In addition antioxidant activity, total carotenoids, and chlorophyll content were determined in edible leaves. Reducing sugars, β-carotene, total antioxidants capacity, total carotenoids, and chlorophyll content were significantly affected by environmental conditions. The location at lower altitude and closer to the coastline showed high evapotranspiration, thus reducing sugar content, antioxidant activity, and phytonutrients in both storage roots and leaves. Absence of water stress in agro-ecological locations further inland and at higher altitudes was associated with an increase in these compounds. Free radical scavenging activity of DPPH was higher in the storage roots (610.49 µmoles TE/100g) than in the leaves (426.06 µmoles TE/100g); nevertheless, opposite results were found for the ferric ion reducing activity (FRAP). The deep orange-fleshed cultivar A45 contained high β-carotene (15 mg/100g), which is enough to meet RDA for vitamin A. There is evidence of agro-ecological effect on sweet potato nutritional value.

Physicochemical and antioxidative properties of black, brown and red rice varieties of northern Thailand

Rice, the seed of Oryza species, is the major cereal crop in most of the developing countries. Nearly 95% of global rice production is done in Asian countries, and about half of the world’s population consumes it. Some speciality rices are not commonly consumed. Colored rice is one of such variety. In these varieties, high amounts of anthocyanin pigment are deposited in the rice coat to form its black (also known as purple), brown and red colors. Minimum studies are there to explain the properties of these rice varieties of Thailand. Thus, the current study was aimed to assess the physicochemical and antioxidative properties of three rice varieties (Chiang Mai Black rice, Mali Red rice and Suphanburi-1 Brown rice) of different cultivars of northern Thailand. Rice bran extracts of these three cultivars were prepared with different solvents (polar and non-polar) for the evaluation of total phytochemical content and anti-oxidant free-radical-scavenging properties. Chiang Mai Black rice contained higher concentration of phenolic acid, flavonoids, and anthocyanins (Cyanidin 3-glucoside, peonidin 3-glucoside, cyanidin chloride). Chiang Mai Black rice is richer in free-radical-scavenging compounds and activities than the other tested varieties. Polar extractions of rice bran are high in anti-oxidative compounds and activities than non-polar extractions.

The kinetics and induced decomposition on the thermal decomposition of hydroperoxides /

This project is focussed on the thermsLl decomposition of t-butyl hydroperoxide and sec-butyl hydroperoxide at 120°C to 160°C in three alcohol solvents. These are methanol, ethajiol and isopropyl alcohol. The aim of the project was to examine the process of induced decomposition. Thermal decomposition of t-hutyl hydroperoxide and sec-butyl hydroperoxide indicate that these reactions have first-order kinetics with activation energies on the order of 20 to 28 K cal/mole, Styrene was used as a free radical trap to inhibit the induced decomposition. The results permitted calculation of how much induced decomposition occurred in its absence. The experimental resvilts indicate that the induced decomposition is important for t-butyl hydroperoxide in alcohol solvents, as shown by both the reaction rate suid product studies. But sec-butyl hydroperoxide results show that the concerted mechanism for the interaction of two sec-butylperoxy radicals occurs in addition to the induced decomposition. Di-sodium E.D,T.A. was added to reduce possible effects of trace transition metal ion .impurities. The result of this experiment were not as expected. The rate of hydroperoxide decomposition was about the same but was zero-order in hydroperoxide concentration.

Modifications to the Suzuki reaction and mechanistic insights on the NBS mediated cleavage of benzylidene acetals /

One of the most challenging tasks for a synthetic organic chemist today, is the development of chemo, regio, and stereoselective methodologies toward the total synthesis of macromolecules. r . The objective of my thesis was to develop methodologies towards this end. The first part of my project was to develop highly functionalized chirons from D-glucose, a cheap, chiral starting material, to be utilized in this capacity. The second part of the project dealt with modifying the carbon-carbon bond forming Suzuki reaction, which is utilized quite often as a means of combining molecular sub units in total synthesis applications. As previously stated the first area of the project was to develop high value chirons from D-glucose, but the mechanism of their formation was also investigated. The free radical initiated oxidative fragmentation of benzylidene acetals was investigated through the use of several test-case substrates in order to unravel the possible mechanistic pathways. This was performed by reacting the different acetals with N-bromosuccinimide and benzoyl peroxide in chlorobenzene at 70^C in all cases. Of the three mechanistic pathways discussed in the literature, it was determined, from the various reaction products obtained, that the fragmentation of the initial benzylic radical does not occur spontaneously but rather, oxidation proceeds to give the benzyl bromide, which then fragments via a polar pathway. It was also discovered that the regioselectivity of the fragmentation step could be altered through incorporation of an allylic system into the benzylidene acetal. This allows for the acquisition of a new set of densely functionalized. chiral, valuable synthetic intermediates in only a few steps and in high yields from a-Dglucose. The second part of the project was the utilization of the phosphonium salt room temperature ionic liquid tetradecyltrihexylphosphonium chloride (THPC) as an efficient reusable medium for the palladium catalyzed Suzuki cross-coupling reaction of aryl halides, including aryl chlorides, under mild conditions. The cross-coupling reactions were found to proceed in THPC containing small amounts of water and toluene using potassium phosphate and 1% Pd2(dba)3. Variously substituted iodobenzenes, including electron rich derivatives, reacted efficiently in THPC with a variety of arylboronic acids and afforded complete conversion within 1 hour at 50 ^C. The corresponding aryl bromides also reacted under these conditions with the addition of a catalytic amount of triphenylphosphine that allowed for complete conversion and high isolated yields. The reactions involving aryl chlorides were considerably slower, although the addition of triphenylphosphine and heating at 70 ^C allowed high conversion of electron deficient derivatives. Addition of water and hexane to the reaction products results in a triphasic system in which the top hexane phase contained the biaryl products, the palladium catalyst remained fully dissolved in the central THPC layer, while the inorganic salts were extracted into the lower aqueous phase. The catalyst was then recycled by removing the top and bottom layers and adding the reagents to the ionic liquid which was heated again at 50 ^C; resulting in complete turnover of iodobenzene. Repetition of this procedure gave the biphenyl product in 82-97% yield (repeated five times) for both the initial and recycled reaction sequences.