Genotoxicity of coke-oven and urban air particulate matter in in vitro acellular assays coupled with 32P-postlabeling and HPLC analysis of DNA adducts

This study is an in vitro part of the ongoing biomarker studies with population from a polluted region of Northern Bohemia and coke-oven workers from Czech and Slovak Republics. The aim of this study is to compare DNA adduct forming ability of chemical compound classes from both the urban and coke-oven extractable organic mass (EOM) of airborne particles. The crude extracts were fractionated into seven fractions by acid-base partitioning and silica gel column chromatography. In in vitro acellular assays we used calf thymus DNA (CT DNA) with oxidative (+S9) and reductive activation mediated by xanthine oxidase (+XO) under anaerobic conditions. Both the butanol and nuclease P1 versions of 32P-postlabeling for detection of bulky aromatic and/or hydrophobic adducts were used. The results showed that the spectra of major DNA adducts resulting from both the in vitro assays are within the fractions similar for both the urban and coke-oven samples. The highest DNA adduct levels with S9-activation were detected for the neutral aromatic fraction, followed by slightly polar and acidic fractions for both samples. With XO-mediated metabolism, the highest DNA adduct levels were detected for both the acidic fractions. Assuming additivity of compound activities, then the acidic fraction, which in the urban sample comprises a major portion of EOM mass (28%), may contain the greatest activity in both in vitro assays (39 and 69%, +S9 and +XO, respectively). In contrast, the aromatic fraction constituting only 8% of total urban EOM mass may account for comparable activity (34%) with organic acids. The highest DNA adduct forming activity of the coke-oven sample accounts for the aromatic fraction (82 and 63%, +S9 and +XO, respectively) that also contains the greatest portion of the total EOM (48%). To characterize some of the specific DNA adducts formed, we coupled TLC on 20x20 cm plates with HPLC analysis of 32P-postlabeled adducts. In both S9-treated samples of the aromatic fraction, we tentatively identified DNA adducts presumably diolepoxide-derived from: 9-hydroxy-benzo[a]pyrene (9-OH-B[a]P), benzo[a]pyrene-r-7,t-8-dihydrodiol-t-9,10-epoxide[+/-] (anti-BPDE), benzo[b,j,k]fluoranthenes (B[b]F, B[j]F, B[k]F), chrysene (CHRY), benz[a]-anthracene (B[a]A) and indeno[cd]pyrene (I[cd]P). These DNA adducts accounted for about 57% of total DNA adducts detected in both S9-treated samples of the aromatic fraction. DNA adducts of XO-treated samples were sensitive to nuclease P1 and HPLC profiles of the major adducts were markedly different from the major adducts of S9-treated samples. However, the combination of TLC and HPLC did not confirm the presence of DNA adducts derived from 1-nitropyrene (1 NP), 9-nitroanthracene (9 NA) and 3-nitrofluoranthene (3 NF) that were detected by GC-MS in the slightly polar fraction. We concluded that the chemical fractionation procedure facilitates the assessing of DNA adduct forming ability of different chemical compound classes. However, based on the results obtained with the whole extracts, it does not fulfil a task of the actual contribution of individual fractions within the activity of the whole extracts. Our results are the first in detecting of DNA adducts derived from urban air and coke-oven particulate matter.     

Rapid shift in peak melatonin secretion associated with improved performance in short shift work schedule

OBJECTIVE: To describe the changes in the characteristics of human immunodeficiency virus (HIV)-related deaths in children with perinatally acquired infection. METHODS: A retrospective review of all deaths that occurred in HIV-infected children managed at The New York Hospital-Program for Children with AIDS during a 7-year period from January, 1990, to December, 1996. Differences in the characteristics at death between 15 children who died in 1990 and 10 children who died in 1996 were analyzed. RESULTS: Fifty-eight deaths in our cohort of HIV-infected children were identified during the 7-year period. The mean age at death was 4.43 years. Sixty-nine percent of children were black, 55% were male and 94% were receiving Medicaid. The mean weight/age Z score was -3.9 and the mean CD4 index was 0.067 with 65% having <50 CD4 cells/microl at the time of death (TOD). The most common organ/organ systems to be involved at the TOD were lung (78%) and central nervous system (61%). Mycobacterium avium complex (MAC) was the most common isolate at the TOD (26%) followed by Pneumocystis carinii (20%) and Pseudomonas aeruginosa (17%). The leading non-infectious cause of death was cardiac failure (9%). Comparison of the characteristics at the TOD between 1990 and 1996 revealed significant differences in mean age (2.1 vs. 9.2 years, P < 0.0001), mean CD4 count index (0.18 vs. 0.02, P < 0.03), mean number of organ/organ system involvement (3.9 vs. 5.9, P < 0.05), percent receiving antiretroviral therapy (33% vs. 70%, P < 0.02), mean number of years receiving antiretroviral therapy (0.88 vs. 3.86 years, P < 0.01), percent receiving P. carinii pneumonia prophylaxis (27% vs. 100%, P < 0.001), percent receiving MAC prophylaxis/therapy (0% vs. 100%, P < 0.0001), and cause of death from P. carinii pneumonia (53% vs. 0%, P < 0.01). CONCLUSIONS: Compared with children who died in 1990, HIV-infected children who died in 1996 were significantly older, more lymphopenic and more likely to have a greater number of organ system involvements and to have received antiviral therapy and antimicrobial prophylaxis. In 1996 no child died of P. carinii pneumonia. In 1996 MAC and P. aeruginosa were the two most important opportunistic infections causing death. These changes in the characteristics at death will warrant review of resources used in treating these children and may be critical in advising parents and care givers about the prognosis of this chronic infection.     

Effect of filtration on virgin olive oil stability during storage

The effect of filtration and dehydration on the stability and quality of virgin olive oil during storage at room temperature (25 °C) and under accelerated conditions (40 °C) was studied. Different types of monovarietal olive oil, namely unfiltered (UF), filtered (F) and filtered‐dehydrated (FD), were obtained from Arbequina, Colombaia, Cornicabra, Picual and Taggiasca cultivars. Results showed that filtration and dehydration decreased the rate of hydrolysis of the triacylglycerol matrix, especially at the higher temperature and in oils with a higher initial free acidity (e.g. free acidity of 0.82% and 0.63% in UF and FD Colombaia samples, respectively, after 8 months of storage), and delayed the appearance of rancid defects (e.g. UF and FD Arbequina samples lost extra‐virgin grade after 10 and 12 months of storage, respectively). The formation of simple phenols due to the hydrolysis rate of their secoiridoid derivatives was also greater in unfiltered olive oils (e.g. 174 μmol/kg and 137 μmol/kg in UF and FD Picual samples, respectively, after 8 months of storage). Thus, filtration and especially dehydration could help to prolong the shelf life of high‐quality and less stable olive oils like those obtained from the Arbequina and Colombaia varieties.     

Characterisation of flavour compounds in Biska – a herbal spirit produced with mistletoe

Mistletoe (Viscum album) is mostly used in its herbal form owing to its content of diverse biologically active substances. In Croatia's Adriatic region, it is used for production of Biska, a strong alcoholic beverage produced by maceration of mistletoe in spirit or brandy, with or without the addition of honey. In this work, Biska was characterised by analysis of aroma compounds and physicochemical parameters and is the first study of the aroma profile of a strong alcoholic beverage produced with mistletoe. The aroma of 14 samples of Biska – five commercial and nine homemade – was analysed using GC/MS with solid‐phase microextraction. A total of 166 aroma compounds were detected. Major components were ethyl esters (medium and long chain fatty acids), fatty alcohols, isopropyl myristate, aldehyde decanal and some terpenes. The distribution of aroma compounds was evaluated by principal component analysis. Some significant differences in physicochemical properties were observed including ethanol concentration (28–44% v/v) and pH (4.4–6.5). Although the colour of Biska was light brown with greenish reflections, there were differences in chromatic parameters. The commercial samples were darker and were less transparent than homemade Biska, perhaps reflecting more extensive maceration/extraction or the addition of caramel. © 2018 The Institute of Brewing & Distilling     

Molecular and Morphological Characterization of Immiscible SAN/EPDM Blends Filled by Nano Filler

The compatibilizing effect of nano sized calcium carbonate filler on immiscible blends of styrene‐co‐acrylonitrile/ethylene propylene diene (SAN/EPDM) was examined. The surface energy of the calcium carbonate was modified by stearic acid. The compatibility of SAN/EPDM blends was studied by following the glass transition temperature T_g by DSC. SEM was used to observe the blend morphology and the X‐ray analyzer was used to detect the calcium from filler in samples. Mechanical properties of the blends were determined, and related to changes of polymer‐filler interactions and morphology. The results suggest that the morphology of the SAN/EPDM blends studied was affected by the reduction of surface energy of the filler.

SEM micrograph of an SAN/EPDM blend with 5% of maximally treated filler.     

Influence of calcium carbonate filler and mixing type process on structure and properties of styrene–acrylonitrile/ethylene–propylene–diene polymer blends

The properties of styrene–acrylonitrile (SAN) and ethylene–propylene–diene (EPDM) blends containing different types of calcium carbonate filler were studied. The influence of mixing type process on the blend properties was also studied. Two different mixing processes were used. The first one includes mixing of all components together. The other process is a two‐step mixing procedure: masterbatch (MB; EPDM/SAN/filler blend) was prepared and then it was mixed with previously prepared polymer blend. Surface energy of samples was determined to predict the strength of interactions between polymer blend components and used fillers. The phase morphology of blends and their thermal and mechanical properties were studied. From the results, it can be concluded that the type of mixing process has a strong influence on the morphological, thermal, and mechanical properties of blends. The two‐step mixing process causes better dispersion of fillers in blends as well as better dispersion of EPDM in SAN matrix, and therefore, the finest morphology and improved properties are observed in blends with MB. It can be concluded that the type of mixing process and carefully chosen compatibilizer are the important factors for obtaining the improved compatibility of SAN/EPDM blends. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012     

Fluctuations in the Levels of Antioxidant Compounds and Antioxidant Capacity of Ten Small Fruits During One Year of Frozen Storage

The fluctuations in antioxidant capacity (2,2-diphenyl-1-picrylhydrazyl, 2,2´-azinobis (3-ethylbenzthiazoline-6-sulphonic acid), and ferric reducing/antioxidant power assays), total phenol, total flavonoid, and total anthocyanin contents of 10 small fruits (sweet cherries, sour cherries, strawberries, red currants, raspberries, blackberries, hawthorn, cornelian cherries, and red and white grapes) were monitored during storage at ?20°C. After one year in storage, all varieties except hawthorn and white grapes retained equal or slightly greater total phenol content in comparison to the initial values. The total flavonoid and total anthocyanin levels also remained stable or even increased after 12 months in all fruits exept hawthorn and strawberries. Red currants and raspberries exhibited optimal preservation of antioxidant capacity, while hawthorn showed a decrease in the antioxidant capacity during different time points in storage as well as at the end of storage. Analyzed fruits were classified into four main groups based on hierarchical cluster analysis.     

The surface energy as an indicator of miscibility of SAN/EDPM polymer blends

Surface properties of blends prepared of styrene-acrylonitrile and ethylene-propylene-diene changing the homopolymer proportions and compatibilized by high impact polystyrene have been studied by contact angle measurement. The surface free energy of interphase of homopolymers pairs, work of adhesion, and wetting coefficient were calculated using Wu’s geometric mean method and the total surface free energy and acid–base components of the blends by using van Oss, Good and Chaudhury method. Blends were also characterized by dynamic mechanical analysis and by scanning electron microscopy. The miscibility of studied blends was estimated through changes of surface free energy, energy of interface and through the shifts of glass transition temperature and changes in morphology. From the results, it can be seen that added compatibilizer reduces the interface energy and provides more homogenous system by interfacial segregation and rearrangements of molecules at the blend surface. The results of morphological observations reveal that the addition of a small percentage of compatibilizer decreases the domain size of the dispersed phase and enhances the compatibility of the blends.     

No evidence for an association of AChR beta-subunit gene (CHRNB1) with myasthenia gravis

Using a polymorphic dinucleotide repeat, we have investigated the contribution of the gene encoding the beta-subunit of the muscle acetylcholine receptor (CHRNB1), the target autoantigen, to the susceptibility to myasthenia gravis (MG). We have combined a case-control study (comparing 143 patients and 162 controls) and a transmission-disequilibrium test bearing on 35 simplex families with heterozygous parents. There was no evidence for an association of CHRNB1 with MG, even after subgrouping patients according to thymus histology, or other clinical criteria. Interestingly however, the shortest four variants of the CHRNB1 microsatellite were seen only in patients with thymus hyperplasia and in none of the control subjects (P < 0.0025).