An optimized FAIRE procedure for low cell numbers in yeast

We report an optimized low‐input FAIRE‐seq (Formaldehyde‐Assisted Isolation of Regulatory Elements‐sequencing) procedure to assay chromatin accessibility from limited amounts of yeast cells. We demonstrate that the method performs well on as little as 4 mg of cells scraped directly from a few colonies. Sensitivity, specificity and reproducibility of the scaled‐down method are comparable with those of regular, higher input amounts, and allow the use of 100‐fold fewer cells than existing procedures. The method enables epigenetic analysis of chromatin structure without the need for cell multiplication of exponentially growing cells in liquid culture, thus opening the possibility of studying colony cell subpopulations, or those that can be isolated directly from environmental samples.     

Reactivity of partially reduced arylhydroxylamine and nitrosoarene metabolites of 2,4,6-trinitrotoluene (TNT) toward biomass and humic acids

Sequential anaerobic/aerobic treatment of 2,4,6-trinitrotoluene (TNT) generally results in the incorporation of residues into biomass and natural organic matter fractions of a system. To better understand the potential contribution of hydroxylamine and nitroso moieties in these reactions, studies were conducted using model systems taking advantage of the biocatalytic-activity of Clostridium acetobutylicum that does not produce aminated TNT derivatives. To evaluate binding to biomass only, systems containing cell-free extracts of C. acetobutylicum and molecular hydrogen as a reductant were employed. At the end of treatment, mass balance studies showed that 10% of the total 14C was associated with an insoluble protein-containing precipitate that could not be extracted with organic solvents. Model reactions were conducted between a mixture of 2,4-dihydroxylamino-6-nitrotoluene (DHA6NT) and 4-hydroxylamino-2,6-dinitrotoluene (4HADNT) and 1-thioglycerol to test the involvement of the nitroso-thiol reaction in binding to biomass. It was demonstrated that DHA6NT formed a new and relatively polar product with 1-thioglycerol only in the presence of oxygen. The oxygen requirement confirmed that the nitroso functionality was responsible for the binding reaction. The reactivity of arylhydroxylamino and nitrosoarene functionalities toward International Humic Substance Society (IHSS) peat humic acid was evaluated under anaerobic and aerobic conditions, respectively. 4HADNT showed no appreciable reactivity toward peat humic acid. Conversely, the nitrosoarene compound, nitrosobenzene, showed rapid reactivity with peat humic acid (50% removal in 48 h). When tested with two other humic acids (selected on the basis of their protein content), it became apparent that the proteinaceous fraction was responsible at least in part for the nitrosoarene's removal from solution. Furthermore, the pretreatment of the humic acids with a selective thiol derivatizing agent had a considerable effect on their ability to react with nitrosobenzene. Finally, molecular modeling tools were used to compare the electrophilic characteristics of potential nitroso intermediates forming from the oxidation of arylhydroxylamino metabolites of TNT. Molecular modeling analysis demonstrated that the more reduced TNT derivative containing nitroso groups were more likely to react with nucleophiles in humic substances than the less reduced nitroso intermediates.     

Randomised trial investigating effect of a novel nicotine delivery device (Eclipse) and a nicotine oral inhaler on smoking behaviour, nicotine and carbon monoxide exposure, and motivation to quit

OBJECTIVE: To monitor the effect of a novel nicotine delivery device that may produce fewer carcinogens (Eclipse) on cigarette smoking, carbon monoxide and nicotine concentrations, and motivation to give up smoking. The smoker's own brand of cigarette and a nicotine replacement product (Nicotrol inhaler) were used as comparisons. DESIGN: After baseline data were recorded, smokers were randomised to either Eclipse or inhaler for two weeks and then switched to the other product for another two weeks. Thereafter a second baseline was obtained. SETTING AND PARTICIPANTS: Fifty smokers were included and data are reported for the 40 with complete data sets. The smokers were not trying to quit but were interested in trying a new product to reduce their risk. They visited a smoking clinic 10 times during the six week period of the trial. INTERVENTION: No counselling to aid reduction by Eclipse or inhaler was given. MAIN OUTCOME MEASURES: At each visit smoking status and carbon monoxide concentrations were recorded. In half of the visits withdrawal symptoms, attitudes towards smoking, heart rate, and blood nicotine concentrations were also recorded. RESULTS: Eclipse use decreased the number of cigarettes smoked per day (cpd) from 19.1 cpd at baseline to 2.1 cpd (p < 0.001), but increased carbon monoxide concentrations in parts per million (ppm) from 21.0 ppm to 33.0 ppm (p < 0.001). A similar decrease in cigarettes smoked per day was seen with the Nicotrol inhaler, from 19.1 cpd to 4.8 cpd (p < 0.001), but carbon monoxide decreased from 21.0 ppm to 12.7 ppm (p < 0.001). The blood nicotine concentration remained fairly stable with Eclipse, increasing slightly from 16.8 ng/ml to 18.0 ng/ml, while for the inhaler a significant drop was noted, from 16.8 ng/ml to 12. 2 ng/ml (p < 0.002). Craving and withdrawal did not increase with Eclipse. Few significant adverse events occurred with Eclipse. CONCLUSIONS: Eclipse can dramatically decrease cigarette consumption without causing withdrawal symptoms or decreases in nicotine concentrations or motivation to quit altogether. Unlike the inhaler, Eclipse produces an increase in carbon monoxide concentration. Thus Eclipse may not be a safer cigarette.     

The relationship between sensory quality and volatile compounds in raw juice processed from elderberries (Sambucus nigra L.)

Elderberry juice was processed by pressing of 101 samples of elderberries (Sambucus nigra L.) picked in Austria, England and Denmark at optimum maturity. Taste panels trained in determination of elderberry flavour evaluated the juices and the results from the sensory evaluation were correlated to the content of volatile compounds. The volatile compounds were isolated from elderberry juices by dynamic headspace technique and analysed by GC-FID and GC-MS. A total of 59 volatile compounds were identified, 9 of which have not previously been detected in elderberry products. A total of 53 compounds occurred in quantifiable amounts and significant differences among the investigated elderberry samples were observed for 50 compounds. The importance of the isolated volatile compounds to elderberry flavour was determined by classical factor analysis using the results from the sensory evaluation and information on the odour of the individual volatile compounds. Classical factor analysis were used including determination of eigenvalues, factor loadings and factor scores by varimax rotation of standardised variables for extraction of four non correlated factors (principal components) explaining approximately 73% of the variation in elderberry flavour. Twenty-eight volatile compounds and sensory quality were included in the four factors. Using the compounds with highest numerical values (loadings) the four factors were named: fresh-elderberry-elder flower (35.2%), fresh-fruity-sweet (17.3%), fresh-fruity (13.7%) and green-floral-fruity (6.7%). Three compounds with characteristic elderberry odour (-damascenone, nonanal, dihydroedulan) were closely correlated with the sensory quality of elderberry together with compounds with fruity, floral, green and/or miscellaneous odours.     

The effectiveness of ozone and acidulant treatments in extending the refrigerated shelf life of fresh-cut potatoes

Abstract: The objective of the study was to determine the effectiveness of acidulant dip treatments (with or without aqueous ozone) to reduce enzymatic browning and to extend the shelf life of fresh‐cut potato slices during refrigerated storage (4 °C) for 28 d. Potato slices subjected to aqueous ozone (2 ppm) had significantly (P≤ 0.05) higher L‐values and lower a‐values, but ozone did not appear to have any effect on aerobic plate counts (APCs) or polyphenol oxidase (PPO) activity. NatureSeal (NS) and sodium acid sulfate (SAS) were the most effective acidulant treatments in reducing browning (significantly [P≤ 0.05] higher L‐values, lower a‐values, and browning index values) regardless of ozone treatment. NS and SAS also had lower PPO activity compared to other treatments on days 0 and 28, and significantly (P≤ 0.05) lower APCs (≤2.00 log CFU/g) over refrigerated storage. Therefore, the SAS treatment was comparable to NS, a commercially available product, and showed promise as an effective antibrowning dip to reduce browning and spoilage in fresh‐cut potato products. Practical Application: A 1% SAS dip treatment which included 1% citric and 1% ascorbic acid was found to be an effective antibrowning dip for fresh‐cut potatoes along with NatureSeal®'s PS‐10, compared to other treatments. They were both effective in maintaining low microbial counts over refrigerated storage. Additionally, aqueous ozone washes (2 ppm) showed significant benefits to reduce browning; however, ozone did not affect microbial counts or PPO enzyme activity. Therefore, the SAS treatment could have potential use in the fruit and vegetable industry to reduce browning and spoilage in fresh‐cut potato products.     

Fat deposition, fatty acid composition and meat quality: A review

This paper reviews the factors affecting the fatty acid composition of adipose tissue and muscle in pigs, sheep and cattle and shows that a major factor is the total amount of fat. The effects of fatty acid composition on meat quality are also reviewed. Pigs have high levels of polyunsaturated fatty acids (PUFA), including the long chain (C20-22) PUFA in adipose tissue and muscle. The full range of PUFA are also found in sheep adipose tissue and muscle whereas cattle ‘conserve’ long chain PUFA in muscle phospholipid. Linoleic acid (18:2n − 6) is a major ingredient of feeds for all species. Its incorporation into adipose tissue and muscle in relation to the amount in the diet is greater than for other fatty acids. It is deposited in muscle phospholipid at a high level where it and its long chain products eg aracidonic acid (20:4n − 6) compete well for insertion into phospholipid molecules. Its proportion in pig adipose tissue declines as fat deposition proceeds and is an index of fatness. The same inverse relationships are not seen in ruminant adipose tissue but in all species the proportion of 18:2n − 6 declines in muscle as fat deposition increases. The main reason is that phospholipid, where 18:2n − 6 is located, declines as a proportion of muscle lipid and the proportion of neutral lipid, with its higher content of saturated and monounsaturated fatty acids, increases. Oleic acid (18:1cis − 9), formed from stearic acid (18:0) by the enzyme stearoyl Co-A desaturase, is a major component of neutral lipid and in ruminants the same enzyme forms conjugated linoleic acid (CLA), an important nutrient in human nutrition. Like 18:2n − 6, -linolenic acid (18:3n − 3) is an essential fatty acid and is important to ruminants since it is the major fatty acid in grass. However it does not compete well for insertion into phospholipid compared with 18:2n − 6 and its incorporation into adipose tissue and muscle is less efficient. Greater biohydrogenation of 18:3n − 3 and a long rumen transit time for forage diets also limits the amount available for tissue uptake compared with 18:2n − 6 from concentrate diets. A positive feature of grass feeding is that levels of the nutritionally important long chain n − 3 PUFA are increased ie EPA (20:5n − 3) and DHA (22:6n − 3). Future research should focus on increasing n − 3 PUFA proportions in lean carcasses and the use of biodiverse pastures and conservation processes which retain the benefits of fresh leafy grass offer opportunities to achieve this. The varying fatty acid compositions of adipose tissue and muscle have profound effects on meat quality. Fatty acid composition determines the firmness/oiliness of adipose tissue and the oxidative stability of muscle, which in turn affects flavour and muscle colour. Vitamin E is an essential nutrient, which stabilises PUFA and has a central role in meat quality, particularly in ruminants.     

The Influence of Ethanol on the Foaming Properties of Beer Protein Fractions: A Comparison of Rudin and Microconductivity Methods of Foam Assessment

The effect of ethanol on the foaming properties of beer protein fractions was studied using a microconductivity method and nitrogen gas to generate the foam. Increasing the ethanol concentration resulted in a decrease in foam stability. Interfacial studies including thin film drainage and dilational elasticity measurements indicated that ethanol reduced the rigidity of the adsorbed protein layer resulting in accelerated drainage from the foam lamellae and increased probability of film rupture. These results conflict with data from the Rudin method (using nitrogen gas to generate the foam) which indicate that, at low concentration, ethanol improves foam stability. These apparently conflicting results may be explained by the foam positive effects of a decline in bubble size and increase in bulk viscosity observed for the Rudin method, contrasted with the negative influence of a reduction in surface viscosity observed for the microconductivity foam assessment method.     

Caring for a family member with a traumatic brain injury

The responses to a questionnaire on subjective burden are reported for 52 primary caregivers of a group of persons with traumatic brain injuries sustained an average of 6 years previously. The aim of the study was to examine satisfaction with social support, perception of coping skills, and appraisal of symptoms as predictors of strain in the carers. A range of responses, both positive and negative, to the work of caring for a relative with a head injury was reported. A high prevalence rate of emotional and behavioural changes in the persons with head injuries was found and the amount of distress caused by these symptoms was found to be predictive of burden. The other factor important in predicting burden was the carers' ratings of their satisfaction with their ability to cope with the work of caregiving. Social support, injury severity, and the demographic characteristics of the persons with head injury and their carers were not significant predictors. Depression in the carers was also investigated and the variable most predictive of elevated depression scores was coping satisfaction. These findings reinforce the importance of strengthening carers coping resources in rehabilitation work with head injured persons and their families.