Effects of addition of strawberry juice pre- or postfermentation on physiochemical and sensory properties of fermented goat milk.

Fruit juice addition can influence the physiochemical and sensory properties of fermented milk. Effects of adding strawberry juice (15% vol/vol) pre- or postfermentation on particle size, rheology properties, volatile flavor profile, and microstructure of fermented goat milk were investigated. Samples with juice added prefermentation showed larger reinforcement in particle size, apparent viscosity, and storage and loss moduli than samples with juice added postfermentation. Compared with the control, all samples showed improved fruit aroma and reduced goaty flavor; this effect was more remarkable in samples in which juice was added before fermentation. Fermented goat milk with strawberry juice added prefermentation showed the highest levels of 1-hexanol and 2-hexen-1-ol (26.16 and 22.03%, respectively) and the lowest percentage of octanoic acid (2.47%), which are mainly responsible for fruit and goaty flavor, respectively. Microstructure analysis showed that samples with juice added prefermentation had a compact protein network. Results indicated that addition of strawberry juice before fermentation may be a good technology for developing a stirred strawberry fermented goat milk.

Interactions between whey protein or polymerized whey protein and soybean lecithin in model system.

Soybean lecithin is often used as a surfactant in food formulation. The aim of this study was to investigate the interactions between soybean lecithin (SL, 0-3%, wt/vol) and whey protein (WP, 10%, wt/vol) or polymerized whey protein (PWP, 10%, wt/vol) induced by heating WP solutions at 85°C for 0 to 20 min at pH 7.0. The samples were evaluated for zeta potential, particle size, morphology, rheological properties, thermal properties, secondary structure, and surface hydrophobicity. Zeta potential of WP increased linearly as SL level increased from 0 to 3%, whereas that of PWP changed with plateau at SL level of 1%, which may be due to the aggregation of SL. The addition of SL increased the particle size and apparent viscosity of both WP and PWP. All the samples exhibited different morphology depending on SL level and heating time according to transmission electron microscopy images. Whey protein showed obviously decreased gelation time and increased storage modulus in the presence of SL. Differential scanning calorimetry curves confirmed the effects of SL on the thermal properties of both WP and PWP. Circular dichroism spectra indicated that SL had effects on the secondary structure of both WP and PWP. The changes in surface hydrophobicity indicated the hydrophobic interactions between WP/PWP and SL. Data indicate that the physicochemical and functional properties of WP and PWP can be altered by adding soybean lecithin.

Effects of gamma radiation on microbial, physicochemical, and structural properties of whey protein model system.

Gamma radiation has been used in food processing for many years, though it has certain effects on food components. Whey protein solutions (10%/30%, wt/vol) were treated with gamma radiation at various dosages (10-25 kGy) and evaluated for microbial changes in the solutions and physicochemical and structural changes of whey proteins. Whey protein solutions after gamma radiation showed substantially lower populations of all viable microorganisms than those of controls. The 10% whey protein solution treated at radiation of 20 or 25 kGy remained sterile for up to 4 wk at room temperature. Gamma radiation increased viscosity and turbidity and decreased soluble nitrogen of whey protein solutions compared to nonradiated control samples regardless of radiation dosage. Nonreducing sodium dodecyl sulfate-PAGE suggested that whey proteins under gamma radiation treatment formed aggregates with high molecular weights. Reducing sodium dodecyl sulfate-PAGE showed that disulfide bonds played a role in gamma radiation-induced whey protein cross-linking. Scanning and transmission electron microscopy micrographs exhibited large aggregates of whey proteins after gamma radiation treatment. Results suggested that gamma radiation could be applied to whey protein solution for purposes of reducing microbial counts and cross-linking protein molecules.

Expression and antibody generation of the cancer-testis antigen, BIOT2-S.

Biot2-S is a mouse cancer-testis antigen gene that was identified using the cross-reactive serological analysis of recombinant cDNA expression libraries (SEREX) technique in the State Key Laboratory of Biotherapy, West China Hospital, Sichuan University. To express BIOT2-S and generate its antibody for further investigation, the Biot2-S prokaryotic recombinant expression vector Biot2-S/pGEX6P-1 was constructed with Escherichia coli DH5α as a cloning vector, and BIOT2-S was expressed in E. coli Rosetta (DE3). The recombinant BIOT2-S was expressed in the form of an inclusion body and the targeted recombinant BIOT2-S was produced at the level of approximately 25% total bacterial proteins after being induced with optimum conditions (0.2 mM isopropyl-ß-D-thiogalactopyranoside for 6 h at 37°C). The target protein was purified by glutathione S-transferase (GST)-trap FF affinity chromatography and detected by western blot. The purified recombinant protein was further confirmed by electrospray ionization quadrupole time-of-flight mass spectrometry after removal of the GST-tags. Then the purified BIOT2-S was used to immunize adult rabbits to generate its antibody. The antibody was purified and its specificity determined. The titer of the antibody was shown to reach 10(4) and the antibody was demonstrated to be able recognize the corresponding protein in the testes of mouse and chicken; the tumor cell lines CT-26 and S180 also reacted with the antibody. This study provides a valuable foundation for further research on the cancer-testis antigen BIOT2-S.

Key autophagic targets and relevant small-molecule compounds in cancer therapy.

Autophagy is a highly conserved lysosomal degradation process which can recycle unnecessary or dysfunctional cell organelles and proteins, thereby playing a crucial regulatory role in cell survival and maintenance. It has been widely accepted that autophagy regulates various pathological processes, among which cancer attracts much attention. Autophagy may either promote cancer cell survival by providing energy during unfavourable metabolic circumstance or can induce individual cancer cell death by preventing necrosis and increasing genetic instability. Thus, dual roles of autophagy may determine the destiny of cancer cells and make it an attractive target for small-molecule drug discovery. Collectively, key autophagy-related elements as potential targets, oncogenes mTORC1, class I PI3K and AKT, as well as tumour suppressor class III PI3K, Beclin-1 and p53, have been discussed. In addition, some small molecule drugs, such as rapamycin and its derivatives, rottlerin, PP242 and AZD8055 (targeting PI3K/AKT/mTORC1), spautin-1, and tamoxifen, as well as oridonin and metformin (targeting p53), can modulate autophagic pathways in different types of cancer. All these data will shed new light on targeting the autophagic process for cancer therapy, using small-molecule compounds, to fight cancer in the near future.

UNC51-like kinase 1, autophagic regulator and cancer therapeutic target.

Autophagy, the cell process of self-digestion, plays a pivotal role in maintaining energy homoeostasis and protein synthesis. When required, it causes degradation of long-lived proteins and damaged organelles, indicating that it may play a dual role in cancer, by both protecting against and promoting cell death. The autophagy-related gene (Atg) family, with more than 35 members, regulates multiple stages of the process. Serine/threonine protein kinase Atg1 in yeast, for example, can interact with other ATG gene products, functioning in autophagosome formation. One mammalian homologue of Atg1, UNC-51-like kinase 1 (ULK1) and its related complex ULK1-mAtg13-FIP200 can mediate autophagy under nutrient-deprived conditions, by protein-protein interactions and post-translational modifications. Although specific mechanisms of how ULK1 and its complex transduces upstream signals to the downstream central autophagy pathways is not fully understood, past studies have indicated that ULK1 can both suppress and promote tumour growth under different conditions. Here, we summarize some properties of ULK1 which can regulate autophagy in cancer, which may shed new light on future cancer therapy strategies, utilizing ULK1 as a potential new target.

Solubility and relative absorption of copper, iron, and zinc in two milk-based liquid infant formulae.

The relative solubility of copper, iron, and zinc in two experimental liquid infant formulae are examined. The results of these trials suggest that substituting organic forms of copper and iron in the mix results in an almost three-fold increase in their solubility (iron lactate, 73.4% vs ferrous sulfate, 27.6% and copper gluconate, 11.3% vs cupric sulfate, 3.0%). Organic zinc substitutes did not show this pattern of increased solubility Electron microscopy was employed to document the changes in protein-protein and protein-lipid interactions and to examine the pattern of electron-dense precipitates in the two experimental formulae. Electron micrographs of the liquid infant formula that had been formulated using inorganic salts (sulfates) showed extensive attachment of denatured whey proteins and casein micelles to the oil droplet surfaces and the surface of the oil droplets were also punctuated by electron-dense granule. The oil droplets of the formula produced using organic versions of the mineral salts were smooth and clear of electron-dense deposits. Experiments were designed to determine whether the observed changes in solubility and microstructure were correlated with increases in relative absorption of the minerals. We applied a technique of in vitro acidification followed by a peptic digestion of the two experimental infant formulae with human milk samples as controls. Coupling this in vitro digestion with an absorption model consisting of live isolated intestinal loops from guinea pig we were able to assess the relative absorption of copper, iron and zinc in the test digests. The relative absorption of the three minerals from digests of human milk was significantly higher than for either of the experimental formulae. Relative mineral absorption from digests of the two experimental infant formulae tested was only significantly different (P < 0.05) for Fe. Based on the results from this study we can conclude that substituting organic forms of iron in bovine milk-base infant formulae would have beneficial effects on both the solubility and bioavailability of this important micronutrient.

Effect of milk preacidification on low fat mozzarella cheese: II. Chemical and functional properties during storage.

The effect of milk preacidification on cheese manufacturing, chemical properties, and functional properties of low fat Mozzarella cheese was determined. Four vats of cheese were made in 1 d using no preacidification (control), preacidification to pH 6.0 and pH 5.8 with acetic acid, and preacidification to pH 5.8 with citric acid. This process was replicated four times. Modifications in the typical Mozzarella manufacturing procedures were necessary to accommodate milk preacidification. The chemical composition of the cheeses was similar among the treatments, except the calcium content and calcium as a percentage of protein were lower in the preacidified treatments. During refrigerated storage, the chemical and functional properties of low fat Mozzarella were affected the most by milk preacidification to pH 5.8 with citric acid. The amount of expressible serum, unmelted cheese whiteness, initial unmelted hardness, and initial apparent viscosity were lower with preacidification. The reduction in initial unmelted cheese hardness and initial apparent viscosity in the pH 5.8 citric treatments represents an improvement in the quality of low fat Mozzarella cheese that allows the cheese to have better pizza bake characteristics with shorter time of refrigerated storage.

Whiteness change during heating and cooling of Mozzarella cheese.

Whiteness (L-value) changes in low-fat and low-moisture, part-skim Mozzarella cheeses during heating (7 to 60 degrees C) and cooling (60 to 7 degrees C) were evaluated. In low-fat Mozzarella, a large increase in whiteness was observed during heating, and a decrease in whiteness was observed during cooling. In low-moisture, part-skim Mozzarella, the whiteness changes during heating and cooling were smaller. Serum phase was removed from low-fat and low-moisture, part-skim Mozzarella cheeses. White protein gels were formed when the isolated serum phase from either low-fat or low-moisture, part-skim Mozzarella was heated. The white gel that formed was composed predominantly of casein and casein proteolysis products. The gel might have been produced by heat-induced, hydrophobic protein-protein interactions, and it tended to dissociate when cooled. Formation of a gel during heating increased light scattering, which increased the L-value. The gel dissociated during cooling and no longer scattered light, which decreased the L-value. We hypothesized that a gel, which was reversible, formed in the serum phase of cheese during heating and might have been responsible for the observed changes in the L-value of low-fat Mozzarella cheese during heating and cooling. The additional fat in low-moisture, part-skim Mozzarella compared with low-fat Mozzarella masked some of the color changes in the serum phase of low-moisture, part-skim Mozzarella. A model was developed to describe the contributions of the casein matrix plus serum phase of Mozzarella cheese and the contribution of fat to the changes in whiteness of Mozzarella cheese during heating and cooling.

Susceptibility of beta-lactoglobulin and sodium caseinate to proteolysis by pepsin and trypsin.

The susceptibility of beta-LG and sodium caseinate to proteolysis by pepsin and trypsin was investigated using SDS or urea-PAGE. The effects were studied of heat, urea, and 2-mercaptoethanol on proteolysis. Native beta-LG was resistant to hydrolysis by pepsin or trypsin because of its compact globular structure. Heat treatment of beta-LG solutions at 90 to 100 degrees C for 5 or 10 min caused changes in the structure or conformation of the protein that rendered it accessible to pepsin and enhanced the extent of proteolysis by trypsin. The susceptibility of beta-LG to proteolysis by pepsin was markedly increased in the presence of urea (3 to 6 M), and the effect was reversible after removal of urea by dialysis. Proteolysis by trypsin was also increased by the presence of 2% 2-mercaptoethanol. Sodium caseinate was very accessible to pepsin without pretreatment and was extensively hydrolyzed at pH 1 to 5 in the presence of 5 M urea (which prevented the protein from precipitation in the isoelectric region); optimal pH was about 2. The activity of pepsin on sodium caseinate at pH 2 was not significantly affected by urea concentration up to about 8 M. The results indicated that the changes in conformation and structure of beta-LG that were induced by heating, reduction, or urea rendered the protein susceptible to peptic hydrolysis.

Effect of individual caseins on plasminogen activation by bovine urokinase-type and tissue-type plasminogen activators.

The effect was examined of individual caseins on the rate of plasminogen activation by bovine urokinase-type and tissue-type plasminogen activators. All individual caseins (alpha-CN, beta-CN, and kappa-CN) enhanced the activity of both types of plasminogen activators. Optimal concentrations for alpha-CN and beta-CN were 5 and 25 micrograms/ml, respectively. The enhancement of enzymatic activity declined when concentrations of alpha-CN and beta-CN were higher. In contrast, increasing concentrations of kappa-CN from 0 to 200 micrograms/ml resulted in corresponding increases in activity of both types of plasminogen activators. On a weight basis, alpha-CN was the most effective enhancer of plasminogen activator activity. Indirect evidence obtained with experiments utilizing alpha-CN immobilized on agarose suggested that the effect is related to extensive binding of plasminogen and both types of plasminogen activators to casein.

Distribution of plasminogen activator in different fractions of bovine milk.

The type and relative amounts of plasminogen activator (PA) in different fractions of bovine milk obtained from 15 Holstein cows were examined. Raw milk was centrifuged to separate skim milk and a somatic cell pellet. PA was mainly localized within the casein fraction, being 42 times that in the serum, and in association with somatic cells. The predominant form of PA in milk casein was isolated from SDS-PAGE gel extracts and had a molecular mass of approximately 75 kDa. Its activity was increased 4.1-fold (P < 0.01) in the presence of fibrin but was unaffected by the presence of amiloride, indicating that it was due to tissue-PA. The predominant forms of PA associated with milk somatic cells were isolated from SDS-PAGE gel extracts and had molecular masses of approximately 30 and approximately 50 kDa. The activity of both proteins was unaffected by the presence of fibrin but was dramatically reduced by the presence of amiloride, indicating that they represented urokinase-PA.