Search results for: ames%20test

Authors: Özlem Arpacı, Zeliha Soysal, Nuran Diril

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Benzoxazoles are heterocyclic compounds that have a fused benzene and an oxazole ring. These heterocyclic compounds are reported to have antibacterial, antitubercular, antifungal, antiviral, antioxidant and anticancer activities. In this study, some benzoxazole derivatives that have broad antimicrobial and potent antitumoral activities, are investigated their mutagenic activities with using the Ames Test. The Ames test was conducted using Salmonella typhimurium TA98, TA100 and TA102 tester strains in the standard plate incorporation assay in the absence of liver S9 fraction. The results are evaluated using SPSS and none of the benzoxazole derivatives showed mutagenic activity using the Ames test in the absence of S9 liver fraction.

Keywords: benzoxazoles, ames test, mutagenic activity, antimutagenic activity, antitumoral activity

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Authors: Awatif Boumaza, Abderraouf Hilali, Hayat Talbi, Houda Sbayou

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The methanolic extract of Zygophyllum cornutun coss, an Algerian medicinal plant, was screened to the presence of mutagenic activity and genotoxic effect using the Ames test (Salmonella/microsome) and the micronucleus assay respectively. Positive results were obtained with both tests. The Ames test showed mutagenic activity in the presence of microsomal activation, while negative result was observed without microsomal activation. In the micronucleus test, two parameters were evaluated: the frequency of the micronucleus that increased in a dose dependent way and the proliferation index that decreased according to the micronucleus frequency. Even that further studies must be carried out, the mutagenic activity and the genotoxic effect of Zygophyllum cornutum should be taken in consideration when used as therapeutic plant.

Keywords: ames test, micronucleus test, mutagenic activity, genotoxicity, Zygophyllum cornutum

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Authors: Nur Fathin Alia Che Wahab, Thirumulu Ponnuraj Kannan, Zuliani Mahmood, Ismail Ab. Rahman, Hanafi Ismail

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Locally produced Biphasic Calcium Phosphate (BCP) consists of hydroxyapatite (HA) and β-tricalcium phosphate (β-TCP) which is a promising material for dentin and bone regeneration as well as in tissue engineering applications. The study was carried out to investigate the mutagenic effect of locally produced BCP using Ames test. Mutagenicity was evaluated with and without the addition of metabolic activation system (S9). This study was performed on Salmonella typhimurium TA98, TA102, TA1537, and TA1538 strains using preincubation assay method. The doses tested were 5000, 2500, 1250, 625, 313 µg/plate. Negative and positive controls were also included. The bacteria were incubated for 48 hours at 37 ± 0.5 °C. Then, the revertant colonies were counted. Data obtained were evaluated using non-statistical method. The mean number of revertant colonies in strains with and without S9 mix treated with locally produced BCP was less than double when compared to negative control for all the tested concentrations. The results from this study indicate that the locally produced BCP is non-mutagenic under the present test conditions.

Keywords: ames test, biphasic calcium phosphate, dentin regeneration, mutagenicity

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Authors: Pravaree Phuneerub, Chanida Palanuvej, Nijsiri Ruangrungsi

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Cymbopogon nardus Rendel (Family Gramineae) is commonly known as citronella grass. The dried root of C. nardus is used for antipyretic, anti-inflammation, anti-analgesic and anticancer in traditional Thai medicine. Transverse sectional and pulverized C. nardus root were illustrated. The volatile oil was extracted from oil gland by hydrodistillation and analysed by GC/MS. Cymbopogon nardus root was exhaustively extracted by continuously maceration in ethanol and water respectively. The mutagenic and antimutagenic properties of the ethanol extract and fractionated water extract of C. nardus root were evaluated by Ames assay using the S. typhimurium strains TA98 and TA100 as the models. The result indicated that the anatomical character of root transverse section displayed epidermis, parenchyma, oil gland, phloem, xylem vessel, endodermis and pith. Histological characters of root powder showed parenchyma containing oleoresin, parenchyma in longitudinal view, reticulate vessel, annular vessel, starch granules and fragment of fiber. The root volatile oil was rich in sesquiterpenes dominated by elemol (22.87%) and alpha-eudesmol (16.09%). For mutagenic activity, the both extracts of C. nardus were no mutagenic toward S. typhimurium strains TA98 and TA100. Furthermore, the ethanol extract and fractionated water extract of C. nardus root demonstrated strong antimutagenic effect against of nitrite treated 1-aminopyrene to S. typhimurium strains TA98 and TA100. This present investigation suggested that the dried root extract of C. nardus can be further developed as promising antimutagenic agent.

Keywords: Cymbopogon nardus, volatile oil analysis, mutagenic, antimutagenic effect, Ames Salmonella assay

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Authors: T. Schilirò, S. Bonetta, S. Bonetta, E. Ceretti, D. Feretti, I. Zerbini, V. Romanazzi, S. Levorato, T. Salvatori, S. Vannini, M. Verani, C. Pignata, F. Bagordo, G. Gilli, S. Bonizzoni, A. Bonetti, E. Carraro, U. Gelatti

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Air pollution is one of the most important worldwide health concern. In the last years, in both the US and Europe, new directives and regulations supporting more restrictive pollution limits were published. However, the early effects of air pollution occur, especially for the urban population. Several epidemiological and toxicological studies have documented the remarkable effect of particulate matter (PM) in increasing morbidity and mortality for cardiovascular disease, lung cancer and natural cause mortality. The finest fractions of PM (PM with aerodynamic diameter <2.5 µm and less) play a major role in causing chronic diseases. The International Agency for Research on Cancer (IARC) has recently classified air pollution and fine PM as carcinogenic to human (1 Group). The structure and composition of PM influence the biological properties of particles. The chemical composition varies with season and region of sampling, photochemical-meteorological conditions and sources of emissions. The aim of the MAPEC (Monitoring Air Pollution Effects on Children for supporting public health policy) study is to evaluate the associations between air pollution and biomarkers of early biological effects in oral mucosa cells of 6-8 year old children recruited from first grade schools. The study was performed in five Italian towns (Brescia, Torino, Lecce, Perugia and Pisa) characterized by different levels of airborne PM (PM10 annual average from 44 µg/m3 measured in Torino to 20 µg/m3 measured in Lecce). Two to five schools for each town were chosen to evaluate the variability of pollution within the same town. Child exposure to urban air pollution was evaluated by collecting ultrafine PM (PM0.5) in the school area, on the same day of biological sampling. PM samples were collected for 72h using a high-volume gravimetric air sampler and glass fiber filters in two different seasons (winter and spring). Gravimetric analysis of the collected filters was performed; PM0.5 organic extracts were chemically analyzed (PAH, Nitro-PAH) and tested on A549 by the Comet assay and Micronucleus test and on Salmonella strains (TA100, TA98, TA98NR and YG1021) by Ames test. Results showed that PM0.5 represents a high variable PM10 percentage (range 19.6-63%). PM10 concentration were generally lower than 50µg/m3 (EU daily limit). All PM0.5 extracts showed a mutagenic effect with TA98 strain (net revertant/m3 range 0.3-1.5) and suggested the presence of indirect mutagens, while lower effect was observed with TA100 strain. The results with the TA98NR and YG1021 strains showed the presence of nitroaromatic compounds as confirmed by the chemical analysis. No genotoxic or oxidative effect of PM0.5 extracts was observed using the comet assay (with/without Fpg enzyme) and micronucleus test except for some sporadic samples. The low biological effect observed could be related to the low level of air pollution observed in this winter sampling associated to a high atmospheric instability. For a greater understanding of the relationship between PM size, composition and biological effects the results obtained in this study suggest to investigate the biological effect of the other PM fractions and in particular of the PM0.5-1 fraction.

Keywords: airborne PM, ames test, comet assay, micronucleus test

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Authors: Nijole Savickiene, Antonella Di Sotto, Gabriela Mazzanti, Rasa Starselskyte, Silvia Di Giacomo, Annabella Vitalone

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Plant lectins are non-enzymic and non-immune origin proteins that specifically recognize and bind to various sugar structures and possess the activity to agglutinate cells and/or precipitate polysaccharides and glycoconjugates. The emerging evidences showed that plant lectins contribute not only to tumour cell recognition but also to cell adhesion and localization, to signal transduction, to mitogenic cytotoxicity and apoptosis. Among chitin-binding lectins, the Urtica dioica agglutinin (UDA), which is a complex of different isoforms, has been poorly studied for its biological activity. In this context and according to the increasing interest for lectins as novel antitumor drugs, present paper aimed at evaluating the potential antimutagenic activity of a lectin-like glycoprotein-enriched fraction from aerial part of Urtica dioica L. Aim: to evaluate the potential chemopreventive properties of a protein - lectin fraction from the aerial part of Urtica dioica. Materials and methods: Protein – lectin fraction has been tested for the antimutagenic activity in bacteria (50–800 mg/plate; Ames test by the preincubation method) and for the cytotoxicity on human hepatoma HepG2 cells (0.06–2 mg/mL; 24 and 48 h incubation). Results: Protein – lectin fraction from stinging nettle was not cytotoxic on HepG2 cells up to 2 mg/mL; conversely, it exhibited a strong antimutagenic activity against the mutagen 2-aminoanthracene (2AA) in all strains tested (maximum inhibition of 56.78 and 61% in TA98, TA100, and WP2uvrA strains, respectively, at 800 mg/plate). Discussion and conclusions: Protein – lectin fraction from Urtica dioica L. possesses antimutagenic and radical scavenging properties. Being 2AA a pro-carcinogenic agent, we hypothesize that the antimutagenicity of it can be due to the inhibition of CYP450-isoenzymes, involved in the mutagen bioactivation.

Keywords: lectins, antimutagenicity, chemoprevention, Urtica dioica

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Authors: Saurabh B. Ganorkar, Atul A. Shirkhedkar

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The need for investigations protecting against toxic impurities though seems to be a primary requirement; the impurities which may prove non - toxic can be explored for their therapeutic potential if any to assist advanced drug discovery. The essential role of pharmaceutical analysis can thus be extended effectively to achieve it. The present study successfully achieved these objectives with characterization of major degradation products as impurities for Zileuton which has been used for to treat asthma since years. The forced degradation studies were performed to identify the potential degradation products using Ultra-fine Liquid-chromatography. Liquid-chromatography-Mass spectrometry (Time of Flight) and Proton Nuclear Magnetic Resonance Studies were utilized effectively to characterize the drug along with five major oxidative and hydrolytic degradation products (DP’s). The mass fragments were identified for Zileuton and path for the degradation was investigated. The characterized DP’s were subjected to In-Silico studies as XP Molecular Docking to compare the gain or loss in binding affinity with 5-Lipooxygenase enzyme. One of the impurity of was found to have the binding affinity more than the drug itself indicating for its potential to be more bioactive as better Antiasthmatic. The close structural resemblance has the ability to potentiate or reduce bioactivity and or toxicity. The chances of being active biologically at other sites cannot be denied and the same is achieved to some extent by predictions for probability of being active with Prediction of Activity Spectrum for Substances (PASS) The impurities found to be bio-active as Antineoplastic, Antiallergic, and inhibitors of Complement Factor D. The toxicological abilities as Ames-Mutagenicity, Carcinogenicity, Developmental Toxicity and Skin Irritancy were evaluated using Toxicity Prediction by Komputer Assisted Technology (TOPKAT). Two of the impurities were found to be non-toxic as compared to original drug Zileuton. As the drugs are purposed and repurposed effectively the impurities can also be; as they can have more binding affinity; less toxicity and better ability to be bio-active at other biological targets.

Keywords: UFLC, LC-MS-TOF, NMR, Zileuton, impurities, toxicity, bio-activity

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Authors: Ali Benali Djouher Leila

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Labiovelarization of velar consonants and labials is a very widespread phenomenon. It is attested in all the major northern Berber dialects. Only the Tuareg is totally unaware of it. But, even within the large Berber-speaking regions of the north, it is very unstable: it may be completely absent in certain dialects (such as the Bougie region in Kabylie), and its extension and frequency can vary appreciably between the dialects which know it. Some dialects of Great Kabylia or the Chleuh domain, for example, "labiovélarize" more than others from the same region. Thus, in Great Kabylia, the adjective "large" will be pronounced: amqqwran with the At Yiraten and amqqran with the At Yanni, a few kilometers away. One of the problems with them is deciding whether it is one or two phonemes. All the criteria used by linguists in this kind of case lead to the conclusion that they are unique phonemes (a phoneme and not a succession of two phonemes, / k + w /, for example). The phonetic and phonological criteria are moreover clearly confirmed by the morphological data since, in the system of verbal alternations, these complex segments are treated as single phonemes: agree, "to draw, to fetch water," akwer, "to fly," have exactly the same morphology as as "jealous," arem" taste," Ames, "dirty" or afeg, "steal" ... verbs with two radical consonants (type aCC). At the level of notation, both scientific and usual, it is, therefore, necessary to represent the labiovélarized by a single letter, possibly accompanied by a diacritic. In fact, actual practices are diverse. - The scientific representation of type does not seem adequate for current use because its realization is easy only on a microcomputer. The Berber Documentation File used a small ° (of n °) above the writing line: k °, g ° ... which has the advantage of being easy to achieve since it is part of general typographical conventions in Latin script and that it is present on a typewriter keyboard. Mouloud Mammeri, then the Berber Study Group of Vincennes (Tisuraf review), and a majority of Kabyle practitioners over the last twenty years have used the succession "consonant +" semi-vowel / w / "(CW) on the same line of writing; for all the reasons explained previously, this practice is not a good solution and should be abandoned, especially as it particularizes Kabyle in the Berber ensemble. In this study, we were interested in two velar consonants, / g / and / k /, labiovellarized: / gw / and the / kw / (we adopted the addition of the "w") for the representation for ease of writing in graphical mode. It is a question of trying to characterize these four consonants in order to see if they have different places of articulation and if they are distinct (if these velars are distinct from their labiovellarized counterpart). This characterization is done using locus equations.

Keywords: berber consonants;, labiovelarization, locus equations, acoustical caracterization, kabylian dialect, algerian language

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Authors: Mmbulaheni Ramulondi, Sandy Van Vuuren, Helene De Wet

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The use of plant combinations to treat various medical conditions is not a new concept, and it is known that traditional people do not only rely on a single plant extract for efficacy but often combine various plant species for treatment. The knowledge of plant combinations is transferred from one generation to the other in the belief that combination therapy may enhance efficacy, reduce toxicity, decreases adverse effects, increase bioavailability and result in lower dosages. However, combination therapy may also be harmful when the interaction is antagonistic, since it may result in increasing toxicity. Although a fair amount of research has been done on the toxicity of medicinal plants, there is very little done on the toxicity of medicinal plants in combination. The aim of the study was to assess the toxicity potential of 19 plant combinations which have been documented as treatments of hypertension in northern KwaZulu-Natal by lay people. The aqueous extracts were assessed using two assays; the Brine shrimp assay (Artemia franciscana) and the Ames test (Mutagenicity). Only one plant combination (Aloe marlothii with Hypoxis hemerocallidea) in the current study has been previously assessed for toxicity. With the Brine shrimp assay, the plant combinations were tested in two concentrations (2 and 4 mg/ml), while for mutagenicity tests, they were tested at 5 mg/ml. The results showed that in the Brine shrimp assay, six combinations were toxic at 4 mg/ml. The combinations were Albertisia delagoensis with Senecio serratuloides (57%), Aloe marlothii with Catharanthus roseus (98%), Catharanthus roseus with Hypoxis hemerocallidea (66%), Catharanthus roseus with Musa acuminata (89%), Catharanthus roseus with Momordica balsamina (99%) and Aloe marlothii with Trichilia emetica and Hyphaene coriacea (50%). However when the concentration was reduced to 2 mg/ml, only three combinations were toxic which were Aloe marlothii with Catharanthus roseus (76%), Catharanthus roseus with Musa acuminata (66%) and Catharanthus roseus with Momordica balsamina (73%). For the mutagenicity assay, only the combinations between Catharanthus roseus with Hypoxis hemerocallidea and Catharanthus roseus with Momordica balsamina were mutagenic towards the Salmonella typhimurium strains TA98 and TA100. Most of the combinations which were toxic involve C. roseus which was also toxic when tested singularly. It is worth noting that C. roseus was one of the most frequently used plant species both to treat hypertension singularly and in combination and some of the individuals have been using this for the last 20 years. The mortality percentage of the Brine shrimp showed a significant correlation between dosage and toxicity thus toxicity was dosage dependant. A combination which is worth noting is the combination between A. delagoensis and S. serratuloides. Singularly these plants were non-toxic towards Brine shrimp, however their combination resulted in antagonism with the mortality rate of 57% at the total concentration of 4 mg/ml. Low toxicity was mostly observed, giving some validity to combined use, however the few combinations showing increased toxicity demonstrate the importance of analysing plant combinations.

Keywords: dosage, hypertension, plant combinations, toxicity

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Authors: Johan Anker

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The purpose of this paper is to discuss the nature of gender construction in modern dystopian fiction, the development of this genre in Young Adult Literature and reasons for the enormous appeal on the adolescent readers. A recent award winning South African text in this genre, The Mark by Edith Bullring (2014), will be used as example while also comparing this text to international bestsellers like Divergent (Roth:2011), The Hunger Games (Collins:2008) and others. Theoretical insights from critics and academics in the field of children’s literature, like Ames, Coats, Bradford, Booker, Basu, Green-Barteet, Hintz, McAlear, McCallum, Moylan, Ostry, Ryan, Stephens and Westerfield will be referred to and their insights used as part of the analysis of The Mark. The role of relevant and recurring themes in this genre, like global concerns, environmental destruction, liberty, self-determination, social and political critique, surveillance and repression by the state or other institutions will also be referred to. The paper will shortly refer to the history and emergence of dystopian literature as genre in adult and young adult literature as part of the long tradition since the publishing of Orwell’s 1984 and Huxley’s Brave New World. Different factors appeal to adolescent readers in the modern versions of this hybrid genre for young adults: teenage protagonists who are questioning the underlying values of a flawed society like an inhuman or tyrannical government, a growing understanding of the society around them, feelings of isolation and the dynamic of relationships. This unease leads to a growing sense of the potential to act against society (rebellion), and of their role as agents in a larger community and independent decision-making abilities. This awareness also leads to a growing sense of self (identity and agency) and the development of romantic relationships. The specific modern tendency of a female protagonist as leader in the rebellion against state and state apparatus, who gains in agency and independence in this rebellion, an important part of the identification with and construction of gender, while being part of the traditional coming-of-age young adult novel will be emphasized. A comparison between the traditional themes, structures and plots of young adult literature (YAL) with adult dystopian literature and those of recent dystopian YAL will be made while the hybrid nature of this genre and the 'sense of unease' but also of hope, as an essential part of youth literature, in the closure to these novels will be discussed. Important questions about the role of the didactic nature of these texts and the political issues and the importance of the formation of agency and identity for the young adult reader, as well as identification with the protagonists in this genre, are also part of this discussion of The Mark and other YAL novels.

Keywords: agency, dystopian literature, gender construction, young adult literature

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Authors: David X. Dong, Qingming Zhang, Meng Lu

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Nitrites enter waterways as runoff from croplands and are discharged from many industrial sites. Excessive nitrite inputs to water bodies lead to eutrophication. On-site rapid detection of nitrite is of increasing interest for managing fertilizer application and monitoring water source quality. Existing methods for detecting nitrites use spectrophotometry, ion chromatography, electrochemical sensors, ion-selective electrodes, chemiluminescence, and colorimetric methods. However, these methods either suffer from high cost or provide low measurement accuracy due to their poor selectivity to nitrites. Therefore, it is desired to develop an accurate and economical method to monitor nitrites in environments. We report a low-cost optical sensor, in conjunction with a machine learning (ML) approach to enable high-accuracy detection of nitrites in water sources. The sensor works under the principle of measuring molecular absorptions of nitrites at three narrowband wavelengths (295 nm, 310 nm, and 357 nm) in the ultraviolet (UV) region. These wavelengths are chosen because they have relatively high sensitivity to nitrites; low-cost light-emitting devices (LEDs) and photodetectors are also available at these wavelengths. A regression model is built, trained, and utilized to minimize cross-sensitivities of these wavelengths to the same analyte, thus achieving precise and reliable measurements with various interference ions. The measured absorbance data is input to the trained model that can provide nitrite concentration prediction for the sample. The sensor is built with i) a miniature quartz cuvette as the test cell that contains a liquid sample under test, ii) three low-cost UV LEDs placed on one side of the cell as light sources, with each LED providing a narrowband light, and iii) a photodetector with a built-in amplifier and an analog-to-digital converter placed on the other side of the test cell to measure the power of transmitted light. This simple optical design allows measuring the absorbance data of the sample at the three wavelengths. To train the regression model, absorbances of nitrite ions and their combination with various interference ions are first obtained at the three UV wavelengths using a conventional spectrophotometer. Then, the spectrophotometric data are inputs to different regression algorithm models for training and evaluating high-accuracy nitrite concentration prediction. Our experimental results show that the proposed approach enables instantaneous nitrite detection within several seconds. The sensor hardware costs about one hundred dollars, which is much cheaper than a commercial spectrophotometer. The ML algorithm helps to reduce the average relative errors to below 3.5% over a concentration range from 0.1 ppm to 100 ppm of nitrites. The sensor has been validated to measure nitrites at three sites in Ames, Iowa, USA. This work demonstrates an economical and effective approach to the rapid, reagent-free determination of nitrites with high accuracy. The integration of the low-cost optical sensor and ML data processing can find a wide range of applications in environmental monitoring and management.

Keywords: optical sensor, regression model, nitrites, water quality

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Authors: Tibor Ajtai, Máté Pintér, Noémi Utry, Gergely Kiss-Albert, Andrea Palágyi, László Manczinger, Csaba Vágvölgyi, Gábor Szabó, Zoltán Bozóki

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According to the best knowledge of the authors, here we experimentally demonstrate first, a quantified correlation between the real-time measured optical feature of the ambient and the off-line measured toxicity data. Finally, using these correlations we are presenting a novel methodology for real time characterisation of ambient toxicity based on the multi wavelength aerosol phase photoacoustic measurement. Ambient carbonaceous particulate matter is one of the most intensively studied atmospheric constituent in climate science nowadays. Beyond their climatic impact, atmospheric soot also plays an important role as an air pollutant that harms human health. Moreover, according to the latest scientific assessments ambient soot is the second most important anthropogenic emission source, while in health aspect its being one of the most harmful atmospheric constituents as well. Despite of its importance, generally accepted standard methodology for the quantitative determination of ambient toxicology is not available yet. Dominantly, ambient toxicology measurement is based on the posterior analysis of filter accumulated aerosol with limited time resolution. Most of the toxicological studies are based on operational definitions using different measurement protocols therefore the comprehensive analysis of the existing data set is really limited in many cases. The situation is further complicated by the fact that even during its relatively short residence time the physicochemical features of the aerosol can be masked significantly by the actual ambient factors. Therefore, decreasing the time resolution of the existing methodology and developing real-time methodology for air quality monitoring are really actual issues in the air pollution research. During the last decades many experimental studies have verified that there is a relation between the chemical composition and the absorption feature quantified by Absorption Angström Exponent (AAE) of the carbonaceous particulate matter. Although the scientific community are in the common platform that the PhotoAcoustic Spectroscopy (PAS) is the only methodology that can measure the light absorption by aerosol with accurate and reliable way so far, the multi-wavelength PAS which are able to selectively characterise the wavelength dependency of absorption has become only available in the last decade. In this study, the first results of the intensive measurement campaign focusing the physicochemical and toxicological characterisation of ambient particulate matter are presented. Here we demonstrate the complete microphysical characterisation of winter time urban ambient including optical absorption and scattering as well as size distribution using our recently developed state of the art multi-wavelength photoacoustic instrument (4λ-PAS), integrating nephelometer (Aurora 3000) as well as single mobility particle sizer and optical particle counter (SMPS+C). Beyond this on-line characterisation of the ambient, we also demonstrate the results of the eco-, cyto- and genotoxicity measurements of ambient aerosol based on the posterior analysis of filter accumulated aerosol with 6h time resolution. We demonstrate a diurnal variation of toxicities and AAE data deduced directly from the multi-wavelength absorption measurement results.

Keywords: photoacoustic spectroscopy, absorption Angström exponent, toxicity, Ames-test

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