Effects of highly ripened cheeses on HL-60 human leukemia cells: Antiproliferative activity and induction of apoptotic DNA damage

To establish cheese as a dairy product with health benefits, we examined the multifunctional role of cheeses. In this report, we clarify whether different types of commercial cheeses may possess antiproliferative activity using HL-60 human promyelocytic leukemia cell lines as a cancer model. Among 12 cheese extracts tested, 6 (Montagnard, Pont-l’Eveque, Brie, Camembert, Danablue, and Blue) revealed strong growth inhibition activity and induction of DNA fragmentation in HL-60 cells. Based on the quantification of nitrogen contents in different cheese samples, a positive correlation between the ripeness of various cheeses and their antiproliferative activity tested in HL-60 cells was displayed. Four varieties of Blue cheese ripened for 0, 1, 2, or 3 mo demonstrated that the Blue cheese ripened for a long term was capable of causing the strong suppression of the cell growth and the induction of apoptotic DNA damage as well as nucleic morphological change in HL-60 cells. Collectively, these results obtained suggest a potential role of highly ripened cheeses in the prevention of leukemic cell proliferation.     

Phenolic components from the leaves of Panax ginseng and their effects on HL-60 human leukemia cells

This study investigates phenolic components from the Panax ginseng C.A. Meyer leaves led to the isolation of 3 flavonoids (1–3). Their structures were characterized on the basis of physicochemical properties, nuclear magnetic resonance (NMR) spectra, electron spray ionization-mass spectrometry (ESI-MS) data, and comparison with those in the literature. Of them, sophoraflavonoloside (2) and prunin (3) were found to inhibit the growth of HL-60 human leukemia cells with IC₅₀ in values of 29.8 and 21.4 μM by the induction of apoptosis.     

Effects of goat cheese on Helicobacter pylori activity and gastrointestinal complaints

To investigate whether and how consumption of goats' milk cheese affects Helicobacter pylori activity in the stomach of infected subjects, a randomized, controlled and blind study was performed. Sixty volunteers consumed 4 × 25 g day⁻¹ “Gouda-type” cheese for 3 weeks, prepared from either goats' or cows' milk. The goats' milk cheese was rich in short- and medium-chain fatty acids. Immediately before the start and thereafter every seven days, H. pylori activity was measured by the ¹³C-urea breath test, and gastrointestinal well-being was recorded weekly by validated questionnaires. Neither goats' nor cows' milk cheese had any significant effect on H. pylori activity. Ingestion of both cheeses improved gastrointestinal well-being but had no effect on stool parameters. The difference of the total symptoms score was significant between day 0 and 21 in each dietary groups, but not between the goats' and cows' milk cheese.     

Flavonoids from Rabdosia rubescens exert anti-inflammatory and growth inhibitory effect against human leukemia HL-60 cells

Eight flavonoids, 5,8,4′-trihydroxy-6,7,3′-trimethoxyflavone (1), 5,4′-dihydroxy-6,7,8,3′-tetramethoxyflavone (2), nodifloretin (3), pedalitin, (4), penduletin (5), cirsiliol (6), luteolin (7), and quercetin (8), were isolated from the aerial parts of Rabdosia rubescens. Structures were established on the basis of 1D and 2D NMR and HRMS data. All flavonoids were tested for cytotoxicity in a small panel of cancer cell lines (Hep G2, COLO 205, MCF-7, and HL-60) and anti-inflammatory activity in LPS-treated RAW264.7 macrophage. Among them, compounds (1) and (4) were modestly active for inhibiting nitrite production in macrophage. Compound (2) was demonstrated to be selectively active against HL-60 cells with an IC₅₀ of 7.55 μM. This value is comparable to the IC₅₀ 4.64 μM of doxorubicin, the positive control used in this investigation.     

Erythrodiol, a natural triterpenoid from olives, has antiproliferative and apoptotic activity in HT-29 human adenocarcinoma cells

Erythrodiol is the precursor of pentacyclic triterpenic acids present in Olea Europaea. Although olive oil and some of its constituents are reported to have anticarcinogenic activities, erythrodiol has not been assessed in its cell biological functions in detail. We therefore determined its effects on cell growth and apoptosis in human colorectal carcinoma HT-29 cells. Proliferation, cytotoxicity, and apoptosis were measured by fluorescence-based techniques. Erythrodiol inhibited cell growth with an EC50 value of 48.8 +/- 3.7 microM without any cytotoxic effects in a concentration range up to 100 microM. However, exposure of cells for 24 h to 50, 100, and 150 microM erythrodiol increased caspase-3-like activity by 3.2-, 4.8-, and 5.2-fold over that in control cells. We here demonstrate for the first time that, in colon adenocarcinoma cells, erythrodiol exerts antiproliferative and proapoptotic activity.     

Effect of the lactoperoxidase system on the activity of mesophilic cheese starter cultures in goat milk

The effect of goats’ milk lactoperoxidase (LP) system on the activity of commercially available mesophilic cheese starter cultures was investigated. The growth and acid production of the starter cultures were measured at 2 h intervals for 8 h in goats’ milk kept at 30°C. Most of the starter cultures examined were found to be sensitive to the LP system, but varied in their susceptibility to inhibition. The activity of the mixed starter cultures CHN11, CHN22, CHN19, DCC240 and Flora Danica Normal was strongly inhibited by the LP system. However, the mixed starter culture LL 50C showed resistance to the LP system. The single strain culture Lactococcus lactis subsp. lactis NCDO 605 was inhibited by the LP system. However, the cultures Lactococcus lactis subsp. diacetylactis NCDO 176 and Leuconostoc mesenteroides subsp. cremoris ATCC 33313 were insensitive to the LP system. The results of this study indicate the need for routine screening of starter cultures for resistance to the LP system before using them for cheesemaking from goats’ milk preserved by the LP system.     

Effect of goat milk composition on cheesemaking traits and daily cheese production

Cheese yield is strongly influenced by the composition of milk, especially fat and protein contents, and by the efficiency of the recovery of each milk component in the curd. The real effect of milk composition on cheesemaking ability of goat milk is still unknown. The aims of this study were to quantify the effects of milk composition; namely, fat, protein, and casein contents, on milk nutrient recovery in the curd, cheese yield, and average daily yield. Individual milk samples were collected from 560 goats of 6 different breeds. Each sample was analyzed in duplicate using the 9-laboratory milk cheesemaking assessment, a laboratory method that mimicked cheesemaking procedures, with milk heating, rennet addition, coagulation, curd cutting, and draining. Data were submitted to statistical analysis; results showed that the increase of milk fat content was associated with a large improvement of cheese yield because of the higher recovery of all milk nutrients in the curd, and thus a higher individual daily cheese yield. The increase of milk protein content affected the recovery of fat, total solids, and energy in the curd. Casein number, calculated as casein-to-protein ratio, did not affect protein recovery but strongly influenced the recovery of fat, showing a curvilinear pattern and the most favorable data for the intermediate values of casein number. In conclusion, increased fat and protein contents in the milk had an effect on cheese yield not only for the greater quantity of nutrients available but also for the improved efficiency of the recovery in the curd of all nutrients. These results are useful to improve knowledge on cheesemaking processes in the caprine dairy industry.     

Ultrafiltration of cheese milk: Effect on plasmin activity and proteolysis during cheese ripening

Havarti 45+ cheese was manufactured from milk concentrated 1.8–4.6-fold by ultrafiltration (UF) and from normal milk, and the effect of concentration factor on plasmin activity and subsequent proteolysis in cheese during ripening was examined. There was decreased plasmin activity and a reduced rate of proteolysis of α_S2-casein and β-casein in the UF-cheeses, compared with the corresponding controls, independent of concentration factor. The decreased plasmin activity and slower breakdown of α_S2-casein and β-casein in UF-cheeses compared with traditional cheeses can be partly explained by the inclusion of an increased amount of plasmin inhibitors into the UF-cheeses. However, it is suggested that the differences in plasmin activity and proteolysis arise mainly as a result of inactivation of the plasminogen activation system during UF-concentration, due to a combination of time, temperature and the presence of air in the UF-equipment. The effect of milk treatment on the plasminogen activation system should be further investigated.     

Effect of ripening time on bacteriological and physicochemical goat milk cheese characteristics

Food Science and Biotechnology - Cheese ripening involves lactose metabolism, lipolysis and proteolysis, which are affected by many factors. The aim of this study was to assess changes due to...     

Use of human lysozyme transgenic goat milk in cheese making: effects on lactic acid bacteria performance

Genetically engineered goats expressing elevated levels of the antimicrobial enzyme lysozyme in their milk were developed to improve udder health, product shelf life, and consumer well-being. The purpose of this study was to evaluate the effect of lysozyme on the development of lactic acid bacteria (LAB) throughout the cheese-making process. Raw and pasteurized milk from 7 lysozyme transgenic goats and 7 breed-, age-, and parity-matched nontransgenic controls was transformed into cheeses by using industry methods, and their microbiological load was evaluated. The numbers of colony-forming units of LAB were determined for raw and pasteurized goat milk, whey, and curd at d 2 and at d 6 or 7 of production. Selective plating media were used to enumerate lactococcal species separately from total LAB. Although differences in the mean number of colony-forming units between transgenic and control samples in raw milk, whey, and cheese curd were non-significant for both total LAB and lactococcal species from d 2 of production, a significant decrease was observed in both types of LAB among d 6 transgenic raw milk cheese samples. In pasteurized milk trials, a significant decrease in LAB was observed only in the raw milk of transgenic animals. These results indicate that lysozyme transgenic goat milk is not detrimental to LAB growth during the cheese-making process.     

Short communication: Antimicrobial activity of pequi (Caryocar brasiliense) waste extract on goat Minas Frescal cheese presenting sodium reduction

Sodium chloride reduction in foods is a significant focus of the dairy industry; however, it can interfere with dairy product quality. Thus, researchers have carried out studies on alternatives to maintain dairy product safety when presenting reduced NaCl content, such as natural antimicrobial addition. Caryocar brasiliense (pequi) is a fruit with high phenolic compound concentrations in the pulp and peel and known antioxidant and antimicrobial properties. This study aimed to define the optimum stage for pequi waste extract addition during cheese manufacturing in order to maintain and prolong the shelf life of reduced-sodium goat Minas Frescal cheese. Four different goat Minas Frescal cheese treatments were carried out: control cheese (without extract; CC), pequi extract addition to milk (CM), pequi extract addition to mass (CS), and cheese immersion in pequi extract (CIE). The treatments were subjected to microbiological (Staphylococcus spp., Escherichia coli, Enterobacteriaceae, coliforms and fecal coliforms, Lactococcus spp., and lactic acid bacteria counts), textural (hardness and consistency), and instrumental color (luminosity, yellow intensity, red intensity, chroma, hue angle, and total color change) analyses. No Enterobacteriaceae, Staphylococcus spp., E. coli, or coliforms and fecal coliforms were detected during storage for any of the assessed samples, including CC. Regarding texture, all samples presented a trend for decreasing rigidity during storage. In addition, lower luminosity values were also observed in cheeses produced with added pequi extract (CM, CS, and CIE) when compared with CC. All cheeses produced with added pequi were stable regarding all evaluated parameters; however, pequi extract addition to milk (CM) was shown to be more efficient, leading to higher textural parameters and better microbiological quality during storage. Thus, the CM treatment is the most recommended for pequi waste extract addition during Minas Frescal cheese manufacture.     

Impact of copper on the induction and repair of oxidative DNA damage, poly(ADP-ribosyl)ation and PARP-1 activity

Copper is an essential trace element involved, among other functions, in enzymatic antioxidative defense systems. However, nonprotein bound copper ions have been shown to generate reactive oxygen species. To gain insight into the discrepancy between the protective properties of copper on the one hand and its toxicity on the other hand, we examined the genotoxic effects of CuSO(4) in cultured human cells. Here we report that copper, at cytotoxic concentrations, induces oxidative DNA base modifications and DNA strand breaks. However, at lower noncytotoxic concentrations, copper inhibits the repair of oxidative DNA damage induced by visible light. As a first mechanistic hint, inhibition of H(2)O(2)-induced poly(ADP-ribosyl)ation was identified in cultured cells and further experiments demonstrated a strong inhibition of the activity of isolated poly(ADP-ribose)polymerase-1 (PARP-1) by copper. Bioavailability studies of copper showed a dose-dependent uptake in cells and pointed out the relevance of the applied concentrations. Taken together, the results indicate that copper, under conditions of either disturbed homeostasis or overload due to high exposure, exerts defined genotoxic effects. Hence, a balance needs to be maintained to ensure sufficient uptake and to prevent overload.     

Tetrahydrocurcumin, a major metabolite of curcumin, induced autophagic cell death through coordinative modulation of PI3K/Akt-mTOR and MAPK signaling pathways in human leukemia HL-60 cells

Scope: Autophagy (type II programmed cell death) is crucial for maintaining cellular homeostasis. Several autophagy‐deficient or knockout studies indicate that autophagy is a tumor suppressor. Tetrahydrocurcumin (THC), a major metabolite of curcumin, has been demonstrated with anti‐colon carcinogenesis and antioxidation in vivo. Methods and results: In the present study, we found that treatment with THC induced autophagic cell death in human HL‐60 promyelocytic leukemia cells by increasing autophage marker acidic vascular organelle (AVO) formation. Flow cytometry also confirmed that THC treatment did not increase sub‐G1 cell population whereas curcumin did with strong apoptosis‐inducing activity. At the molecular levels, the results from Western blot analysis showed that THC significantly down‐regulated phosphatidylinositol 3‐kinase/protein kinase B and mitogen‐activated protein kinase signalings including decreasing the phosphorylation of mammalian target of rapamycin, glycogen synthase kinase 3β and p70 ribosomal protein S6 kinase. Further molecular analysis exhibited that the pretreatment of 3‐methyladenine (an autophagy inhibitor) also significantly reduced acidic vascular organelle production in THC‐treated cells. Conclusion: Taken together, these results demonstrated the anticancer efficacy of THC by inducing autophagy as well as provided a potential application for the prevention of human leukemia.     

Sweet potato protein hydrolysates: antioxidant activity and protective effects on oxidative DNA damage

In this study, sweet potato protein (SPP) hydrolysates were prepared by six enzymes (alcalase, proleather FG‐F, AS1.398, neutrase, papain and pepsin). The antioxidant activities and protective effect against oxidative DNA damage of SPP hydrolysates were investigated. Alcalase hydrolysates exhibited the highest hydroxyl radical‐scavenging activity (IC₅₀ 1.74 mg mL^?1) and Fe²⁺‐chelating ability (IC₅₀ 1.54 mg mL^?1) (P < 0.05). Compared with other five hydrolysates, the hydrolysates obtained by alcalase had the most abundant <3‐kDa fractions. In addition, below 3‐kDa fractions of alcalase hydrolysates showed the highest antioxidant activities and protective effects against DNA damage through both scavenging hydroxyl radicals and chelating Fe²⁺, which was probably because of the increase in several antioxidant amino acids, such as His, Met, Cys, Tyr and Phe, as well as the hydrophobic amino acids. The results suggested that enzymatic hydrolysis could be used as an effective technique to produce high value‐added peptides products from SPP.     

玉米蛋白粉中叶黄素和玉米黄素对口腔癌细胞DNA的损伤作用

研究玉米蛋白粉中叶黄素和玉米黄素对人口腔癌KB细胞的作用机制。在人口腔癌KB细胞的细胞培养液中分别加入40μL/L、60μL/L叶黄素和30μL/L、40μL/L玉米黄素,分别培养48h、72h。用单细胞凝胶电泳(SCGE)检测叶黄素和玉米黄素对人口腔癌KB细胞DNA损伤程度。玉米蛋白粉中叶黄素和玉米黄素可引起KB细胞DNA链断裂,细胞尾长、尾DNA百分含量和尾矩显著高于对照组,差异有统计学意义(P<0.01)。叶黄素和玉米黄素能导致人口腔癌细胞KB的DNA损伤,抑制肿瘤细胞生长。     

葛根黄酮提取物对HL-60细胞bc1-2和bax基因表达的影响

目的：探讨葛根黄酮提取物对HL-60细胞bcl—2和bax基因表达的影响。方法：用RT—PCR和蛋白质免疫印迹技术检测葛根黄酮提取物、葛根素和大豆甙元处理HL-60后，对bcl—2和bax基因的mRNA和蛋白表达的影响。结果：葛根黄酮提取物、葛根素和大豆甙元处理HL-60细胞后能够上调bax mRNA表达水平，下调bcl-2蛋白表达和上调Bax蛋白表达，且随着处理时间的延长其诱导下调和上调能力增强。结论：葛根黄酮提取物诱导HL-60细胞凋亡与其下调bcl-2mRNA和蛋白表达水平和其上调bax mRNA和蛋白表达水平有关。     

蜂蜜添加方式对羊酸乳品质及抗氧化活性的影响

在羊乳中添加蜂蜜,研制一款功能性乳制品。分别于羊乳发酵前和发酵后添加枣花蜜,比较不同工艺蜂蜜羊酸乳的发酵速度、感官品质、菌数、持水力、质构特性以及抗氧化活性差异。发酵前加入枣花蜜,有助于缩短羊酸乳凝乳时间,但整体口感稍差,乳酸菌增殖抑制程度较大,持水力下降,硬度、稠度、内聚性和黏性指数有所降低,抗氧化活性则显著提高;而发酵结束后添加枣花蜜,羊酸乳凝乳时间无显著变化,但整体风味和口感、持水力及抗氧化活性均得到有效提高,且乳酸菌增殖抑制程度较小,硬度、稠度、内聚性和黏性指数则明显降低。蜂蜜添加方式对羊酸乳发酵速度、品质及抗氧化活性有显著影响,可根据产品目标选择合适工艺。