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.     

Diagnosis and pathophysiology of paradoxical embolism

A review of the experience with 134 consecutive patients with germinal cell testicular neoplasia indicates that definitively accurate staging of the malignancy at presentation is the single most important prognostic factor. Nearly two-thirds of the patients with all types of germinal malignancies survived or died of other causes and the highest survival rates were seen among patients with earlier stages of seminoma. In recent years patients with all types of germinal malignancies of the testis have been treated by radical retroperitoneal lymphadenectomy with enhancement of survivals. Patients with seminoma and non-seminomatous tumors exhibited increased survival rates with node dissection. In 66 consecutive lymphadenectomies the complication rate was less than 14 per cent, with only 1 death related to the operation. Retroperitoneal lymphadenectomy not only affords therapeutic advantage but also provides an opportunity for accurate surgical staging of disease and allows for rational decision relative to additional treatment, radiation therapy or chemotherapy.     

Effect of dietary CLA supplementation on renal inflammation in diabetic mice

Dietary conjugated linoleic acid (CLA) supplementation was studied for diabetic renal inflammation improvement through modulation of inflammatory responses and oxidative stress. ICR mice were administrated alloxan (180 mg/kg, i.p.) once and fed an AIN-93G diet, or diets containing 0.5 or 1% CLA for 8 weeks. Dietary CLA supplementation did not change body weights or ameliorate fasting blood glucose, blood urea nitrogen, and kidney MDA levels. However, dietary CLA supplementation improved plasma creatinine levels and overall inflammatory response in diabetic nephropathy by modulation of phosphorylated inhibitory kappa B and the protein levels of interleukine-1β, inducible nitric oxide synthetase, and the c-reactive protein without regulation of blood glucose levels. CLA supplementation in diabetic mice did not restore the manganese superoxide dismutase antioxidant defense system. CLA supplementation was beneficial in diabetic nephropathy through regulation of inflammatory response and has potential as an antiinflammatory terapeutic in patients with mild diabetes and diabetic complications.     

A trout (Oncorhynchus mykiss) perfusion model approach to elucidate the role of blood removal for lipid oxidation and colour changes in ice-stored fish muscle

Whole body saline-perfused rainbow trout (Oncorhynchus mykiss) was ice-stored for 4 weeks and compared with unwashed/washed minces from unbled and bled trout in terms of rancid odour, peroxide value (PV), thiobarbituric acid reactive substances (TBARS) and redness loss. Muscle from saline-perfused fish, which had 72% less total haem, was deficient in rancid odour during the whole storage, while bled (54% less haem) and unbled samples developed rancid odour already after ~4 and 2 days; higher intensity without bleeding. PV/TBARS also developed in the order unbled > bled > perfused samples; however, PV/TBARS were not as completely prevented as rancid odour after perfusion. Saline washing (3 × 3 volumes) of unbled mince removed 84% haem and yielded the second most stable sample while saline washing (1 × 1 volumes) destabilised unbled mince, despite 64% haem removal. Concurrent antioxidant removal during washing of minces obviously counteracted the effect of blood removal and washing fish mince with small volumes of solution should be used with great care.     

Heat Curing of Soy Protein Films at Atmospheric and Sub-atmospheric Conditions

ABSTRACT: The effect of heat curing at atmospheric or subatmospheric conditions on selected properties (moisture content, water vapor permeability (WVP), color, tensile strength (TS), elongation (E), and total soluble matter (TSM) content) of cast soy protein isolate films was investigated. Films were heat cured at 85 °C for 6, 12, 18, or 24 h at absolute pressures of 101.3, 81.32, or 61.32 kPa. Heat-cured films had increased (P < 0.05) TS and decreased (P < 0.05) WVP and E compared to control, unheated films. Heat treatment under vacuum reduced the WVP of films faster than heat curing at atmospheric pressure. High TS values, low E values, and low TSM values were also reached within short heating time under vacuum. However, vacuum treatment increased the size and number of cavities in cured films as evidenced by scanning electron micrographs.     

Effect of modified atmosphere packaging on the shelf-life of coated, whole and sliced mushrooms

Whole and sliced fresh mushrooms (Agaricus bisporus) were packaged with PVC wrap or two polyolefins (PD-941 and PD-961) films after coating with CaCl₂ and chitosan. Package gas composition, color, weight loss and maturity were measured during storage at 12 °C and 80%RH. For PD-961, the highest in-package concentration occurred during the first day of storage regardless of treatments, while wrap and PD-941 showed varying degrees in-package concentration with different processes and coatings. The whiteness of whole mushrooms varied significantly with the type of coating, but not with the type of films. The extent of darkening was greater in coated whole mushrooms than in sliced ones. Weight loss occurred in all packages and varied from 3 (g/100 g) to about 7 (g/100 g) after 6 days of storage. Due to a lower permeability, PD-961 packages had the lowest weight loss. The type of packaging films significantly affected the maturity index, where PD-961 most effectively lowered maturity index for both whole and sliced mushrooms, thus extending the shelf-life. The type of coating did not appear to affect maturity index except for the wrap package where chitosan coating markedly lowered the maturity index of sliced mushrooms.     

Processing of Sapota (Sapodilla): Drying

ABSTRACT: The drying characteristics of sapota ( Achras zapota, cv , Kalipatti), a tropical fruit crop, were studied. Fruits were cut in sizes of half, quarter, and 5 mm slices and dried at temperatures of 55, 60, 65, and 70 °C in convection air and vacuum ovens. Moisture content of the fruit reduced from 72 to 78% (w.b) to 8.5 to 12.5% (w.b.) with drying times of 15 to 35 h in a convection air oven and of 14 to 31 h in a vacuum oven. Log and modified log models were fitted for the drying data. Empirical models were developed to predict drying constants as a function of drying temperature and sizes of samples with good correlation. Keywords: sapota, sapodilla, drying     

Measuring oxidative DNA damage and DNA repair using the yeast comet assay

Chromosomal DNA damage can be a result of several processes and agents of endogenous or exogenous origin. These cause strand breaks or oxidized bases that lead to strand breaks, which relax the normally supercoiled genomic DNA and increase its electrophoretic mobility. The extent of DNA damage can be assessed by single cell gel electrophoresis, where the chromosomal DNA migration distance correlates with the extent of DNA damage. This technique has been used for a variety of applications with several organisms, but only a few studies have been reported for Saccharomyces cerevisiae. A possible reason for this absence is that low cellular DNA content could hamper visualization. Here we report an optimization of the comet assay protocol for yeast cells that is robust and sensitive enough to reproducibly detect background DNA damage and oxidative damage caused by hydrogen peroxide. DNA repair was observed and quantified as diminishing comet tail length with time after oxidative stress removal in a process well described by first‐order kinetics with a tail length half‐life of 11 min at 37 °C. This is, to our knowledge, the first quantitative measurement of DNA repair kinetics in S. cerevisiae by this method. We also show that diet antioxidants protect from DNA damage, as shown by a three‐fold decrease in comet tail length. The possibility of assessment of DNA damage and repair in individual cells applied to the model organism S. cerevisiae creates new perspectives for studying genotoxicity and DNA repair. Copyright © 2010 John Wiley & Sons, Ltd.