Interactions between arginine-rich histones and deoxyribonucleic acids. II. Circular dichroism

Circular dichroism (CD) was used to investigate the conformations of arginine-rich histones, H3 (III or f3) and H4 (IV or f2a1), and DNA in the complexes prepared by four different methods: (A) NaCl gradient dialysis with urea; (B) NaCl gradient dialysis without urea; (C) direct mixing in 2.5 x 10(-4) M EDTA, pH 8.0; and (D) direct mixing in 0.01 M sodium phosphate, pH 7.0. Using the CD spectrum of native chromatin as a criterion to judge the closeness of a complex to its native state, it was observed that a complex made by direct mixing at low ionic strength (methods C and D) is better than the ones made by NaCl gradient dialysis with or without urea (methods A and B). It is explained as a result of lack of ordered secondary structures in histones due to the presence of urea in method A or due to nonspecific aggregation in NaCl without urea (method B). Compared with all the earlier reports in literature on the CD of histone-DNA complexes, the CD spectra of arginine-rich histone-DNA complexes prepared by methods C and D are closest to that of native chromatin both in shape and in amplitude. These results imply (a) that arginine-rich histones play an important role in maintaining the conformation of chromatin and (b) that the binding of these two histones to DNA prepared by methods C and D are close to that in native chromatin. Noticeable variation in conformation of free and bound histone and histone-bound DNA has also been observed in histone H3 with one or two cysteine residues, and in reduced or oxidized state even when the complexes were prepared and examined in the same condition. CD spectra of arginine-rich histones in 0.01 M phosphates, pH 7.0, indicate the presence of alpha-helix which could be responsible for a favorable binding of the less basic regions of these histones to DNA under this condition as demonstrated by thermal denaturation (Yu, S. .S, Li H. J., and Shih, T. Y. (1976), Bio-chemistry, the preceding paper in this issue). To preserve or generate alpha-helical structures in histones seems to be a critical step in reconstituting good histone-DNA complexes.     

Comparison of biota-sediment accumulation factors across ecosystems

Sets of biota-sediment accumulation factors (BSAFs) for fish were compared across ecosystems for nonionic organic chemicals. The sets of BSAFs, when plotted against each other, in log-log space, formed linear relationships and demonstrated that the relative scaling or ranking of the individual BSAFs within a set are consistent, if not the same, across ecosystems. This behavior holds for chemicals that either are, or are not, metabolized by fish. These results demonstrate that sets of BSAF values can differ but with parallel shifts in magnitude between ecosystems (for example, all of the BSAFs in the set are uniformly larger in one ecosystem, while in another they all are uniformly smaller) in response to underlying differences in ecosystem conditions and parameters such as trophic level, diet of the organisms, and distribution of the chemical between the sediment and water column.     

Detection of <Emphasis Type="Italic">Campylobacter</Emphasis> colonies using hyperspectral imaging

The presence of Campylobacter in foods of animal origin is the leading cause of bacterially induced human gastroenteritis. Isolation and detection of Campylobacter in foods via direct plating involves lengthy laboratory procedures including enrichments and microaerobic incubations, which take several days to a week. The incubation time for growing Campylobacter colonies in agar media usually takes 24–48 h. Oftentimes the problem is the difficulty of visually differentiating Campylobacter colonies from non-Campylobacter contaminants that frequently grow together with Campylobacter on many existing agars. In this study, a new screening technique using non-destructive and non-contact hyperspectral imaging was developed to detect Campylobacter colonies in Petri dishes. A reflectance spectral library of Campylobacter and non-Campylobacter contaminants was constructed for characterization of absorption features in wavelengths from 400 to 900 nm and for developing classification methods. Blood agar and Campy-Cefex agar were used as culture media. The study found that blood agar was the better culture medium than Campy-Cefex agar in terms of Campylobacter detection accuracy. Classification algorithms including single-band thresholding, band-ratio thresholding and spectral feature fitting were developed for detection of Campylobacter colonies as early as 24 h of incubation time. A band ratio algorithm using two bands at 426 and 458 nm chosen from continuum-removed spectra of the blood agar bacterial cultures achieved 97–99% of detection accuracy. This research has profound implications for early detection of Campylobacter colonies with high accuracy. Also, the developed hyperspectral reflectance imaging protocol is applicable to other pathogen detection studies.     

Survey of bottled drinking waters sold in Canada for chlorate, bromide, bromate, lead, cadmium and other trace elements

Mineral, spring and other bottled drinking waters sold in Canada in the winter of 1995-96 were surveyed for chlorate, bromide, bromate, Cr(VI), Li, B, Al, Mn, Cu, Zn, Sr, Ba, Be, V, Cr, Co, Ni, As, Se, Mo, Ag, Cd, Sb, Tl, Pb, Na, K, Ca and Mg. Chlorate and bromide were determined by ion chromatography (IC) with conductivity detection, Cr(VI) by IC with colorimetric detection, bromate by solvent extraction and gas chromatography (GC), trace elements by inductively coupled plasma mass spectrometry (ICPMS), and Na, K, Ca and Mg by flame atomic absorption spectrometry (FAA). Most chemicals in the 199 samples analysed were well within national and international drinking water guidelines. World Health Organization and/or Canadian drinking water guidelines were exceeded for B (22 samples), Al (9), Cr (1), Mn (5), Ni (1), As (10), Se (24) and Pb (1). Bromate levels are reported for information purposes and are considered as the maximum concentrations in the samples. In three distilled water products, unexpectedly high concentrations of Cu (88-147 µg l^-1) and Ni (16-35 µg l^-1) were found, and a comparison of distilled and non-distilled waters from two of the brands suggested the likely cause to be contamination during the distillation process. Li concentration in one sample was at a therapeutic dose and could pose an overdose risk to individuals on Li medication.     

Cigarette smoking and quitting behaviors among unemployed adults in the United States.

Little is known about factors associated with smoking among the unemployed. This study estimated the prevalence of smoking and examined sociodemographic factors associated with current, former, and successful quitting among unemployed adults aged 18-64. Cross-sectional data on 13,480 participants in the 1998-1999 and 2001-2002 Tobacco Use Supplements to the Current Population Surveys were analyzed. Multivariate logistic regression analyses were used to examine factors associated with study outcomes (current vs. never, former vs. current, successful quitter vs. other former smoker). Among the unemployed, 35% were current smokers and 13% were former smokers. Of the former smokers, 81% quit successfully for at least 12 months. Participants with family incomes of less than US$25,000 were more likely than those with incomes of $50,000 or more to currently smoke (OR=2.13, 95% CI=1.85-2.46). Service workers and blue-collar workers were less likely than white-collar workers to report former smoking. Participants unemployed for 6 months or more were twice as likely as those unemployed for less than 6 months to quit successfully (OR=2.05, 95% CI=1.07-3.95). Unemployed blue-collar workers had a greater odds ratio of successfully quitting than white-collar workers (OR=1.83, 95% CI=1.17-2.87). Smoking rates were high among the unemployed, and quitting behaviors varied by sociodemographic factors and length of unemployment. Studies are needed to examine the feasibility of cessation interventions for the unemployed.     

Effect of biotin supplementation on meat quality of F1 Wagyu/Black Angus feedlot steers of known genotype

Biotin (D-biotin) was supplemented to F1 Wagyu/Black Angus steers fed a wheat-based ration to evaluate the effect on meat quality. One hundred and eight steers of known Wagyu sire lines were assigned to three biotin treatments (0, 10 and 20mg/head/day) with each treatment replicated four times using an unfasted liveweight of 410.5kg (±24.42 SD). Biotin supplementation had no effect (P>0.05) on beef marbling standard at either the 5/6th or 10/11th rib quartering site, 10/11th rib intra-muscular fat percentage, intra-muscular fat fatty acid composition or adipose melting points. Wagyu genotype had an effect (P<0.05) on beef marbling standard and intra-muscular fat percentage at the 10/11th rib, inter-muscular and intra-muscular melting point and fatty acid composition of intra-muscular fat. A significant (P<0.001) but poor correlation existed between beef marbling standard and intra-muscular fat percentage (R(2)=0.198). Total conjugated linoleic acid had a highly significantly (P<0.0001) positive correlation to intra-muscular fat percentage (R(2)=0.446).