Modulation of the antioxidant/pro-oxidant balance,cytotoxicity and antiviral actions of grape seed extracts

Grape seed extracts (GSEs) were investigated in yeast cells harbouring defects in their antioxidant system (regarding the cellular growth and growth recovery from H₂O₂ insult). GSEs antioxidant activity was detected in wild-type and mutant strains Δcta1, Δgsh1 and Δoye2glr1, while pro-oxidant activity in Δsod1 cells was seen. Assessment of proliferation of prostate cancer PC3 and HBV-replicating HepG2 2.2.15 cells treated with GSEs has shown higher cytotoxicity of red grape seed extract (RW) than white grape seed extract (WW) subjective to dose and period of administration. No antiviral effect was detected by measuring the secreted virion particles in HepG2 2.2.15 cells treated with GSEs. The GSEs play a dual antioxidant/pro-oxidant role in vivo according with the cellular antioxidant system deficiencies and exhibit cytotoxic properties in PC3 and HepG2 2.2.15 cell lines, but no antiviral action against HBV.     

Detection of human serum albumin on protein array using scanning tunneling microscopy

Electrical detection of biological binding events, such as protein–protein interaction and DNA hybridization, has emerged as an alternative method to conventional colorimetric and fluorescence based methods. In this study, we demonstrate an electrical detection technique of protein array which can be simply extended for multifunctional measurements and detection of biological binding events on micro-scale array. Micro-contact technique was used for the fabrication of protein chip. The fabricated protein array on Au substrate was characterized by fluorescence microscopy and scanning tunneling microscopy (STM). The chip was designed to investigate immunocomplexes comprised of our model protein, human serum albumin (HSA), corresponding antibody fragments, and Au nanoparticle–antibody conjugates. The peak-like pulse obtained by electrical tunneling current between these complexes and the STM tip varies on the surface density of the bound complexes. Using the electrical detection technique based on STM, 100 fg/mL of HSA could be successfully detected by STM. Importantly, the proposed concept of measurement could allow multiple analyses of analytes at nano-scale array, which is difficult to be analyzed by conventional fluorescence based method.     

The use of bovine serum protein as an oral support therapy following coronavirus challenge in calves

The objective of this experiment was to investigate the therapeutic efficacy of a supplemental bovine serum protein blend fed to calves challenged with virulent coronavirus. Twelve Holstein bull calves (approximately 3 wk of age) were allocated by initial body weight to Control (n = 5) and treated (n = 7) groups. On d 0, all calves were orally challenged with 1 x 10(7) plaque-forming units of virulent coronavirus isolate. Infection was allowed to progress for 24 h before treatment was started. On d 1, treated calves began receiving 160 g of dry bovine serum powder (16 g IgG) mixed into milk replacer powder (67 g) at both an a.m. and p.m. feeding. Control calves received only milk replacer powder (227 g) at both feedings. Response to coronavirus challenge and dietary treatment was monitored prior to a.m. and p.m. feeding by the collection of multiple clinical measures. Fecal consistency was decreased by coronavirus challenge but was not affected by dietary treatment. Mean daily rectal temperature and heart rate were not affected by dietary treatment. Average packed cell volume was higher in treated calves than in control (35.0 and 27.0%). Coronavirus challenge resulted in an immediate increase in respiration rate, decreasing by d 7. Control calves tended to have a greater average respiration rate compared with treated (28.7 vs. 26.8 breaths/min). Treated calves had a higher average feed intake than control (0.57 vs. 0.44 kg/d). These data suggest that bovine-serum supplemented milk replacer may decrease the severity of disease in young calves exposed to coronavirus.     

天然高分子牛血红蛋白对钾离子的吸附研究

用离子选择电极法研究了牛血红蛋白在各种条件下对钾离子的吸附.结果表明,在中性条件下K~ 几乎没有被吸附;随着溶液pH的增加和K~ 浓度的增加,牛血红蛋白对K~ 的吸附量也增加,在一定条件下达到饱和.牛血红蛋白对K~ 的吸附呈可逆性,但蛋白质本身结构,当pH由>11调至<8.5后,呈不可逆变化.如溶液中存在Na~ ,则K~ 的吸附量下降.表明存在竞争吸附.这些实验结果支持了缔合一诱导理论.     

Effect of beta-glucan from oats and yeast on serum lipids

Heart disease is the leading cause of death in the U.S. One way to reduce the risk of developing the disease is to lower serum cholesterol levels by making dietary changes. In addition to reducing intake of total fat, saturated fat, and dietary cholesterol, serum cholesterol can be further reduced by added fiber, especially from sources rich in beta-glucan. In this review, two sources of beta-glucan are described; one source is oats and the other yeast. Their chemical structures and physical properties are compared, and their effect on serum lipid levels is described. Oat beta-glucans are found in various breakfast cereals and snacks. Usually, several servings of these products are required to meet the Food and Drug Administration's claim of reducing the risk of heart disease. The yeast-derived fiber is a more concentrated source of beta-glucan than the oat product. It is currently being tested in a wide variety of food products.     

Thermal unfolding of β‐lactoglobulin, α‐lactalbumin,and bovine serum albumin. A thermodynamic approach

Heat‐treatment is one of the most commonly used processes in food preparation technology. An understanding of the thermodynamics of protein stability and of conformational changes of proteins, acquired through the measurement of the denaturation temperature, is therefore of particular importance. This paper attempts to shed light on the interpretation of recent calorimetric data on the thennal denaturation of bovine β‐lactoglobulin, α‐lactalbumin, and bovine serum albumin by showing that thermodynamic parameters of heat‐induced unfolding, measured by differential scanning calonmetry, are closely related to the prevailing chemical conditions such as pH, concentration of ions, protein purity, and protein concentration.     

Application of microscopical techniques in the study of autolysosome dynamics in PC12 neurites

Autophagy is a principal degradation pathway for the turnover of intracellular proteins or cytoplasmic organelles in response to starvation. During autophagic activation, autophagosomes fuse with lysosomes to form autolysosomes where incorporated materials are degraded. However, the dynamics of autolysosomes in neurites of live cells was still poorly known. In this study, various subsets of microscope were applied to analyse the autophagy induction and highly dynamic transport of autolysosomes in rat PC12 neurites. Beading formation was found in degenerating PC12 neurites under phase contrast light microscope after serum deprivation. The monomeric red fluorescence protein‐green fluorescence protein‐light chain 3–labelled autolysosomes accumulated throughout PC12 neurites after 18 h of serum deprivation as revealed by fluorescence microscope analysis. The single‐membrane autolysosomes were also visualized in PC12 cells under transmission electron microscope. Moreover, fluorescence recovery after photobleaching experiment, which was conducted by confocal laser scanning microscope, demonstrated that autolysosomes were motile vesicles and moved along PC12 neurites during starvation. The directional transport of monomeric red fluorescence protein ‐labelled autolysosomes in neurites was further monitored by a motorized video microscope. Both anterograde and retrograde transport of autolysosomes were observed. In addition, the autolysosomes were precisely mapped by using 2D Gaussian fitting and then their highly dynamic movement was robustly tracked by using multidimensional assignment. Collectively, by using different microscopical techniques, our results confirmed the dynamic transport of autolysosomes in starved PC12 neurites and may provide valuable insight into understanding the biophysical characteristics of autolysosomes in neurites under physiological and pathological conditions.     

Sonocatalytic damage of bovine serum albumin (BSA) under ultrasonic irradiation by mixed TiO2/SiO2 powder

BACKGROUND: In order to effectively damage some biomolecules under ultrasonic irradiation, a mixed TiO₂/SiO₂ powder with high catalytic activity and selectivity was used as a sonocatalyst. RESULTS: The mixed TiO₂/SiO₂ powder heat treated at 450 °C for 30 min was adopted as a sonocatalyst and the damage to BSA molecules under ultrasonic irradiation was assessed. In addition, the effects of such variables as molar ratio of TiO₂ and SiO₂, treatment temperature and time, ultrasonic irradiation time, catalyst amount, solution acidity, ionic nature and strength, ultrasonic irradiation power and D₂O concentration on the damage to BSA molecules were studied by means of UV‐visible and fluorescence spectra. The results showed that the degree of damage was aggravated by an increase in ultrasonic irradiation time, catalyst amount, solution acidity, ultrasonic irradiation power and D₂O concentration, but was reduced by an increase in ionic strength. CONCLUSION: The results indicated that the mixed TiO₂/SiO₂ powder displayed higher activity and selectivity compared with nano‐sized TiO₂ and SiO₂ powders during the sonocatalytic damage of BSA. The extent of the damage decreased in the order TiO₂/SiO₂ > nano‐sized TiO₂ > nano‐sized SiO₂. These results are of great significance for applying sonocatalytic methods to treat tumours. Copyright © 2008 Society of Chemical Industry     

Albumin stabilizes (–)‐epigallocatechin gallate in human serum: Binding capacity and antioxidant property

(–)‐Epigallocatechin gallate (EGCg) is the major component of green tea and is known to show strong biological activity, although it can be easily oxidized under physiological conditions. In this study, we indicate that EGCg is stable in human serum and that human serum albumin (HSA) stabilizes EGCg under aerobic condition. Although EGCg is usually decomposed within 1 h in aqueous solution at neutral pH, EGCg in serum and phosphate buffer (pH 7.4) containing HSA was stable over 1 h, even at neutral and slightly alkaline pH. Under these conditions, EGCg binds to HSA non‐covalently. The sulfhydryl group acts as an antioxidant for EGCg oxidation. Incubation of EGCg with HSA is accompanied by the oxidation of a free sulfhydryl group in HSA. These results suggest that the antioxidant property and the binding capacity of HSA contribute to the stabilization of EGCg in human serum.