Genotoxicity

Introduction: Medicinal plants, such as flaxseeds play an important role in the health of individuals and communities. Regarding the phytochemical properties of flaxseed, this study aimed to investigate the hydroalcoholic effects of this plant on the genetic toxicity of cyclophosphamide in peripheral blood lymphocytes.

Materials & Methods: The flaxseed extract was initially prepared by maceration using ethanol solvent in two rounds. After one hour of incubation with different concentrations, the blood samples were incubated with 750µ M in a bain-marie at 37° C for 24 h. To evaluate the production of micronucleus in dual nuclear lymphocytes suppressed in cytokines, the slides were prepared and evaluated using light microscopy. The data were analyzed in SPSS software and the mean values were compared using Tukey's test and.
Ethics code: 30.96.3.1002

Findings: The incubation of blood samples with cyclophosphamide leads to induced additional genotoxicity in lymphocytes. Moreover, flaxseed extract pretreatment significantly reduced the micronucleus frequency (P<0.0001). In addition, the results showed the effective role of flaxseed extract as a protective agent in reducing the genotoxicity of the pesticide cyclophosphamide.

Discussion & Conclusions: According to the obtained results, flaxseed is a potent antigenotoxic agent against cyclophosphamide-induced DNA damage. Since the flaxseed extract does not have cytotoxic effects, it can be used as a protective agent against the toxic effects of cyclophosphamide.

The study found 280 ug/ kg of acrylamide (carcinogenic with genotoxic potential) in experimental carob syrup samples that were cooked at a constant temperature of 110ºC; less than the 303 ug/kg of the commercial samples. Correlatively, these results indicated that the more time of exposure of solids soluble and water evaporation during a constant temperature cooking, is related to a higher content of acrylamide in carob syrup. The different levels of acrylamide contains in the carob syrup marks depends on the productive method, having a variety range from 0 ug/kg of acrylamide to 303 ug/kg. Additionally, the more concentration of solids caused by the temperature, major is the soluble solids that increase other components different than sugars which contribute to a bitterness flavor in carob syrup. The low per capita consumption and the low frequency of carob syrup consume in the range delimited from 0.7 g/ person/ day contribute to not to pass the values limits of acrylamide intake. The combinated presence of acrylamide and HMF (hydroxymethylfurfural) do not overcome the maximum limit consumption of acrylamide; in other words it is not dangerous (real) for the human health considering the low rate of peruvian intake. There are some commercial samples that indicate an acrylamide result of 0 ug/kg which are probably related to an optimal technical process or the metabolization to another element that are not studied yet. On the other hand, a young person with a lower weight of 63.7 kg, do not should eat more than 25 daily portions of 42 g of carob syrup with a acrylamide content of 0.303 mg/kg (ppm); instead an adult with a 71.3 kg weight can intake 28 daily portions of carob syrup without any associated health risk to acrylamide consumption. Finally, the less contain of acrylamide in carob syrup is associated to a major daily portions of carob syrup consumption. CAA = Amount of acrylamide in algarrobin (ug / kg) * Kg consumption of algarrobin / kg person-day CHMFA = Amount of HMF in algarrobin (ug / kg) * Kg consumption of algarrobin / kg person-day Objectives Quantify the presence of acrylamide and hydroxymethylfurfural (HMF) in the consumption of algarrobina in Piura City-Peru. Methods The carob pod is harvested in a yellow state. The sugars of carob pod are extracted by a leaching process from the carob tree. Correlatively, it is chopped in boiling temperature in natural environmental conditions. Then, the extract is filtered and concentrated to more than 70º Brix in order to obtain soluble solids. Four samples of carob syrup (with different soluble solids) were analyzed by using ion chromatography, mass spectrophotometry and liquid chromatography to quantify the content of acrylamide and HMF. To determine the amount of acrylamide and HMF consumption in carob syrup, it was used the per capita consumption of the surveyed people, specifically the intake for an average person with 72 kg weight. Results For one kilogram of carob, 171.4 ml of algarrobin was obtained, with a density of 1.4 g / ml, a pH: 4.796, 4.26, 5.02 and 5.27 respectively, with a content of soluble solids of 77.07, 81.3, 74.98 and 82.6% and a per-capita consumption of algarrobin of 0.17 lt

In the face of the increasing numbers and haphazard siting of mechanic workshops and the consequent environmental pollution due to its activities, there is paucity of information in the literature on the possible genotoxic and mutagenic effects of the associated waste and the soils of these workshops. This study therefore aimed at assessing the pollution level of the soils from the automobile workshop in Lagos and the genotoxic potential of their simulated leachate using Allium cepa L. 2 kg of soil samples were collected at a depth of 0-15 cm (top soil) using soil auger from five different points at the automobile mechanic workshop in order to get a representative sample (composite sample) and transferred directly into clean, sterile containers. A control soil sample was obtained from the Botanical garden of the University of Lagos, Akoka. The physicochemical analysis of the soil was also carried out using standard methods. The samples were analysed for lead (Pb), chromium (Cr), cadmium (Cd), zinc (Zn), arsenic (As), nickel (Ni), and mercury (Hg) using inductively coupled plasma atomic emission spectroscopy (ICP-AES) after acid digestion. Aliphatic and aromatic hydrocarbons, TPH and oil and grease were determined. Leachate simulation from the soil was carried out according to the American Society for Testing and Materials. The result showed an obvious influence of automobile workshop activities on the physicochemical properties of soil as well as on the root meristem of Allium cepa. Automobile workshop soil contained significantly higher concentrations for all analysed heavy metals (Cd, As, Cu, Pb, Ni and Zn) in comparison to the uncontaminated garden soil. Cd, Cu and Pb levels were higher than specified regulatory standards. All the leachates concentrations showed one form of aberration: sticky chromosomes, C-mitosis, bridges, lagging chromosomes, binucleate, vagrant cells and micronuclei. The highest aberration occurred in 50% concentration of the treatment while the lowest occurred in the control. The aberrations observed varied with exposure time. The results of the present study showed that simulated leachates from an automobile workshop is genotoxic and mutagenic in the bioassays used in this study. The soils

Context: Algae have gained importance in cosmeceutical product development due to their beneficial effects on skin health and therapeutical value with bioactive compounds. Spirulina platensis Parachas (Phormidiaceae) is renowned as a potential source of high-value chemicals and recently used in skincare products.
Objective: This study develops and evaluates skin creams incorporated with bioactive S. platensis extract.
Materials and methods: Spirulina platensis was cultivated, the aqueous crude extract was prepared and in vitro cytotoxicity of S. platensis extract in the range of 0.001–1% concentrations for 1, 3 and 7 d on HS2
keratinocyte cells was determined. Crude extracts were incorporated in skin cream formulation at 0.01% (w/w) concentration and in vitro wound healing and genotoxicity studies were performed.
Immunohistochemical staining was performed to determine the collagen activity.
Results: 0.1% S. platensis extract exhibited higher proliferation activity compared with the control group with 198% of cell viability after 3 d. Skin cream including 1.125% S. platensis crude extract showed
enhanced wound healing effect on HS2 keratinocyte cell line and the highest HS2 cell viability % was obtained with this concentration. The micronucleus (MN) assay results indicated that S. platensis extract
incorporated creams had no genotoxic effect on human peripheral blood cells. Immunohistochemical analysis showed that collagen 1 immunoreactivity was improved by increased extract concentration and it was strongly positive in cells treated with 1.125% extract incorporated skin cream.
Conclusions: The cell viability, wound healing activity and genotoxicity results showed that S. platensis incorporated skin cream could be of potential value in cosmeceutical and biomedical applications.

“Ames” bacterial mutation tests are widely performed for evaluation and registration of new materials including industrial chemicals, agrochemicals, medical devices, pharmaceuticals, pharmaceutical impurities and other materials. Tests are used to predict their potential long-term adverse health effects (including carcinogenicity). Given their importance, pre-screening ‘miniaturized’ versions have been developed
which allow higher throughput and use less test material, including the widely-employed 24-well micro-Ames (mAmes) test which uses 20 times
less material. However, little quantitative information has been published on the methodology or sensitivity of this system. We describe methods
and results used in direct comparisons of the sensitivity of micro and standard systems using the same cultures, formulations, etc. Initial testing utilized the plate incorporation method and, later, the pre-incubation method. In a subsequent phase of testing, a four-way direct comparison was made between the preincubation and plate incorporation methods in both systems using some direct-acting mutagens. Tests used only those strain/S9/chemical combinations
where a response was expected. Historical control results accumulated during testing are also presented. Spontaneous and induced revertant colony counts for the µAmes system were consistently proportionate and approximately 1/20th those for the standard Ames test. Sensitivities of the two systems were found to be nearly identical in almost all cases for a wide variety of weak and strong inorganic and organic mutagens. Standardized procedures and increased reliability of the estimate of the
background revertant frequency in the µAmes system means that the two systems give equivalent results and are expected to be highly predictive of one another.

The importance of Allium cepa test contributes knowledge in preventing toxicity in the environment. This test widely used in determining genotoxic and cytotoxic substances found in the water system. In this study, the genotoxic effects of H 2 O 2 (Hydrogen Peroxide) and CH 2 O (Formalin) were determined using Allium cepa root tip cells while the water samples collected from the mining areas of Sorex Barobo, Surigao del Sur and Rosario, Agusan del Sur were also tested for genotoxicity. One-Way ANOVA shows that water samples treated with H 2 O 2 were statistically significant (P<0.05) when compared to CH 2 O while the water sample collected from the mining areas shows statistically non-significant. A concentration-dependent increased were observed among the dividing cells and aberrant cells of the treated and collected water samples. The observed abnormalities were seen into its roots morphology indicating genotoxicity. Thus, the obtained results in this study show that Allium cepa test is useful as bio-indicator to detect genotoxicity.

This paper reports the first data from an integrated study investigating genotoxicity in the Brantas River, Java, Indonesia. Results showed that organic sediment extracts from the sites in the Brantas Delta retained genotoxic compounds identified using the SOS Chromotest and that the Aloo River and, to a lesser extent, the Surabaya River were the most contaminated studied sites. This genotoxicity was attributable to compounds that did not require any bioactivation under the test conditions. Occurrence of genotoxic effects was further investigated in erythrocytes from Nile tilapia, Oreochromis niloticus. High numbers of micronuclei were counted, especially in fish sampled in the rivers of the Brantas Delta. Moreover, cytoplasmic alterations which could be indicative of the presence of lipofuscin were found in the cytoplasm of the fish blood cells, especially in fish from the Aloo, Surabaya and Kalimas rivers. Altogether, our data showed that genotoxicity is occurring in fish living in rivers of the delta of the Brantas River and suggest that sediments from these sites may constitute a major source of pollution and hazard for species living or feeding in the area.

Abstract
Environmental reservoirs are the potential sources of pathogenic E. coli. These are ideal habitat for survival, growth, and proliferation of E. coli as they are rich in nutrients. A wide range of different animals harbor pathogenic E. coli, which are consistently associated with human and animal diarrheal infection. Infected cattle typically excrete pathogenic E. coli. These discharges are released into the aquatic environment or storm water runoff. The surface water carries a number of pathogenic bacteria when used for the drinking purposes. Apart from these, the water is treated in municipal treatment plants and supplied through pipelines for drinking purposes. The processed clean water sometimes also gets contaminated due to leakage of nearby sewer lines. Highly prevalent diarrheagenic E. coli variants are present in the surface and potable waters in India. The consumption of unsafe water in developing countries causes a number of disease outbreaks. The detection of target pathogens needs improvements to overcome existing drawbacks and should be based on simple, sensitive and specific methodologies. Polymerase Chain Reaction is one of the latest techniques in modern biology which is effective in the detection of these pathogens. Real Time PCR technique has advantages over conventional methods in terms of rapidity, high specificity and sensitivity. The real time PCR uses primer pair and molecular probe (TaqMan, Molecular Beacon, Scorpion etc.) for the sensitive detection of pathogens.
Keywords: E. coli, Water, Detection, PCR, Real Time PCR