Cytotoxic and inflammatory effects of contact lens solutions on human corneal epithelial cells in vitro

Purpose

To ascertain the effect that four contact lens (CL) multipurpose solutions (MPS) have on the viability and release of pro-inflammatory cytokines from human corneal epithelial cells (HCEC).

Cytotoxic effects of 7-O-butyl naringenin on human breast cancer MCF-7 cells

The effect of 7-O-butyl naringenin (BN), a chemically synthesized derivative of naringenin, was tested on the proliferation of human breast cancer MCF-7 cells. BN inhibited the proliferation of MCF-7 cells in dosedependent manner (IC₅₀: 67.5±2.1 μM), resulting in an increase in the sub-G1 phase cell population. BN induced the generation of intracellular reactive oxygen species (ROS), which were reduced by pretreatment with N-acetylcysteine (NAC). BN also increased the phosphorylation of stress-activated protein kinase/c-Jun NH₄-terminal kinase 1/2 (SAPK/JNK1/2), c-Jun, and p38. However, the phosphorylation of extracellular-regulated kinase 1/2 (Erk1/2) was decreased in BN-treated cells. Pretreatment of cells with the specific inhibitors SP600125 and SB203580 diminished the BN-induced activation of SAPK/JNK1/2 and p38, respectively. These results indicate that the BN-induced cytotoxicity of MCF-7 cells is mediated by the generation of ROS as well as through the p38, SAPK/JNK1/2, and c-Jun activation signaling pathways. BN may therefore possess chemotherapeutic potential as an anti-proliferative agent.     

Simultaneous determination of two Monascus metabolites in red yeast rice by HPLC using fluorescence detection

A reversed-phase HPLC method with fluorescence detection for the determination of two Monascus metabolites, monasfluore A (MFA) and monasfluore B (MFB), in red yeast rice was carried out. Optimum conditions for the extraction and chromatographic separation were investigated. The method was validated through the following performance criteria: linearity, stability, limit of detection (LOD), quantification (LOQ), etc. This assay was successfully used for determination of the MFA and MFB in 20 samples inoculated from different Monascus sp. The results revealed that significant variations were demonstrated in the contents of the MFA and MFB in these samples. The high contents of both MFA and MFB in sample 13 were found to be 81.400 and 26.300 mg g^?1, respectively. The low contents of both MFA and MFB in sample 14 were also found to be 0.010 and 0.003 mg g^?1, respectively.     

Optimization of adenoviral production in human embryonic kidney cells using response surface methodology

Adenoviruses are the most commonly used vectors in clinical trials for gene therapy. How to efficiently produce abundant and high-quality adenoviral vectors for therapeutic research is a challenge for biochemical engineers. A recombinant adenovirus carrying a green fluorescent protein (GFP) gene together with an anchorage-dependent 293 cell line is used as a model system for evaluating the effects of chemicals on adenoviral production in this study. Our aim is to develop a formulation to be added to an infection medium that could enhance the in vitro production of adenoviral vectors. Eleven ingredients obtained from a literature survey were screened for their stimulatory effects on adenoviral production using the 50% tissue culture infectious dose (TCID(50)) method. Among these ingredients, sucrose and mannitol when supplemented to the infection medium significantly increased adenovirus titer. Central composite design and response surface methodology were also adopted to determine the optimal concentrations of sucrose and mannitol. The formulation developed, which is composed of DMEM/F12 medium plus 0.54 M sucrose and 0.37 M mannitol, can significantly increase adenoviral production by 13-fold that of the control (DMEM/F12 medium).     

Cranberry derived proanthocyanidins can prevent pathogen invasion of kidney epithelial cells

The in vitro effectivity of cranberry derived proanthocyanidins (PACs) for the mitigation of kidney cell infection by selected uro- and entero-pathogens is examined with an adhesion/invasion assay and confocal microscopy. This study demonstrates that PACs effectively reduce invasion of canine kidney cells by pathogenic bacteria: Escherichia coli CFT073 and O157:H7, Enterococcus faecalis 29212, and Pseudomonas aeruginosa 10145. These effects demonstrate the potential for cranberry derived PACs as a useful tool in the prevention of kidney infection.     

Interactions of lactic acid bacteria with human intestinal epithelial cells: effects on cytokine production

As a participant in the mucosal immune response, the intestinal epithelial cell must respond to a variety of stimuli, including lactic acid bacteria (LAB) consumed in the diet. The objective of this study was to compare the abilities of several strains of LAB to modulate cytokine secretion by human intestinal epithelial cell (IEC) line HT-29. Certain strains of Lactobacillus rhamnosus, Lactobacillus delbrueckii, and Lactobacillus acidophilus suppressed the production of the chemokine RANTES by stimulated HT-29 IEC, although the magnitude of this suppression varied depending on the nature of the bacterial growth medium. Similarly, specific strains showed growth condition-dependent suppression of HT-29 interleukin-8 (IL-8) production. Strain-dependent effects were also seen for the suppression of tumor necrosis factor alpha (TNF-alpha) and transforming growth factor beta (TGF-beta) production. The binding of several of these bacterial strains to the HT-29 cell line was also examined. Different strains were found to have differing abilities to interact with IEC, with L. rhamnosus R0011 being the strain that generally had the most extensive effects on HT-29 cytokine production and also bound to HT-29 IEC most effectively. Modulation of IEC cytokine production has the potential to profoundly affect the mucosal microenvironment, influencing the immune response to pathogens and other ingested antigens.     

Cytotoxic effects of geranyl flavonoid derivatives from the fruit of Artocarpus communis in SK-Hep-1 human hepatocellular carcinoma cells

Breadfruit (Artocarpus communis) is cultivated in tropical and subtropical regions as a traditional starch crop. This study investigated the cytotoxic activity of geranyl flavonoid derivatives isolated from the fruit of A. communis. Two new geranyl flavonoid derivatives, arcommunol A and arcommunol B, together with four known compounds, were isolated from the fruit of A. communis. The effects of these compounds on the viability of HepG2, Hep3B, PLC5 and SK-Hep-1 cells were investigated. Arcommunol A showed the highest inhibitory activity with 2.05 μM IC₅₀ in SK-Hep-1 cells. Treatment with arcommunol A changed the ratio of expression levels of pro- and anti-apoptotic Bcl-2 family members and subsequently induced the activation of apoptosis-inducing factor, apoptotic protease activating factor-1, caspase-9 and caspase-3, which may result in a cleavage of poly(ADP-ribose) polymerase. These findings provide critical information regarding the chemopreventive potential of arcommunol A from the fruit of A. communis.     

桐油甘油三酯对体外培养的人体肿瘤细胞的毒性

以纯度为96%的α-桐酸作为对照,考察了从桐油中经NARP-HPLC分离得到的3组含共轭亚麻酸的甘油三酯对人体肿瘤细胞株ECA-109 (食管癌)、SGC-7901(胃癌)、Bel-7402(肝癌)、LS-174-T(结肠癌)、KB(口腔癌)和正常细胞株 L02(肝细胞)的细胞毒性.结果发现α-桐酸对KB、Bel-7402和SGC-7901细胞的毒性明显弱于3组含共轭亚麻酸的甘油三酯.结果提示,除了α-桐酸的共轭三烯结构是桐油甘油三酯的细胞毒性的物质基础之外,含共轭亚麻酸的甘油三酯的分子结构对其抗肿瘤功效也有重要贡献.     

Cytotoxic impact of phenolics from Lamiaceae species on human breast cancer cells

The aim of this study was to evaluate the cytotoxicity of dried aqueous extracts from Thymus serpyllum (ExTs), Thymus vulgaris (ExTv), Majorana hortensis (ExMh), and Mentha piperita (ExMp), and the phenolic compounds caffeic acid (CA), rosmarinic acid (RA), lithospermic acid (LA), luteolin-7-O-glucuronide (Lgr), luteolin-7-O-rutinoside (Lr), eriodictiol-7-O-rutinoside (Er), and arbutin (Ab), on two human breast cancer cell lines: Adriamycin-resistant MCF-7/Adr and wild-type MCF-7/wt. In the MTT assay, ExMh showed the highest cytotoxicity, especially against MCF-7/Adr, whereas ExMp was the least toxic; particularly against MCF-7/wt cells. RA and LA exhibited the strongest cytotoxicity against both MCF-7 cell lines, over 2-fold greater than CA and Lgr, around 3-fold greater than Er, and around 4- to 7-fold in comparison with Lr and Ab. Except for Lr and Ab, all other phytochemicals were more toxic against MCF-7/wt, and all extracts exhibited higher toxicity against MCF-7/Adr. It might be concluded that the tested phenolics exhibited more beneficial properties when they were applied in the form of extracts comprising their mixtures.     

Anti-inflammatory and proliferative responses in human and ovine ovarian surface epithelial cells

The majority of ovarian cancers (>90%) are believed to derive from the ovarian surface epithelium (OSE); a single layer covering the entire surface of the ovary. At ovulation, the OSE cell layer undergoes an inflammatory response, involving cell death and growth, in order to overcome ovarian surface rupture. Abnormalities during these processes are believed to contribute to the development of tumours. Using primary cultures of OSE cells, we have compared anti-inflammatory and proliferative responses directly between human and ovine OSE cells to further establish the use of ovine OSE cells as a suitable model system for the study of human OSE cells. In order to compare effects of inflammatory stimulation, expression and activity of 11betahydroxysteroid dehydrogenase (11betaHSD) type 1 was measured in OSE cells in response to interleukin (IL)-1alpha. As previously identified in human OSE cells, treatment of ovine OSE cells with IL-1alpha stimulated a concomitant increase of 11betaHSD type 1 mRNA (31-fold; P <0.05) and oxoreductase activity, indicating an increased production of anti-inflammatory cortisol. To compare the growth of human and ovine OSE cells, OSE cell number was measured in response to treatment with gonadotropins or growth factors. In the presence of FSH, LH or human chorionic gonadotropin (hCG), ovine and human OSE cell growth was similarly stimulated >1.2-fold (P <0.05). In the presence of connective tissue growth factor (CTGF) and more significantly insulin growth factor I (IGF-I), human and ovine OSE cell growth was also similarly stimulated >1.2-fold (P <0.05) and >1.5-fold (P <0.01), respectively. The induction of both human and ovine OSE cell growth by IGF-I or hCG was further shown to be dependent on activation of the MAP kinase/extracellular-signal-regulated kinase (ERK) pathway. Stimulation of ovine OSE cell growth by hepatocyte growth factor (HGF) was similarly shown to be ERK-dependent; however, for human OSE cells, HGF only mildly stimulated ERK phosphorylation and failed to stimulate OSE cell growth. The demonstration that human and ovine OSE cells share similarities at the level of cell signalling, gene expression and cellular growth supports the use of ovine OSE cells as a suitable model for the study of human OSE cells.     

Comparative characteristics of rice wine fermentations using Monascus koji and rice nuruk

Food Science and Biotechnology - Wine fermentations using rice media containing either Monascus koji or rice nuruk were performed and fermentative characteristics based on the koji type were...     

Effect of Bacillus cereus exocellular factors on human intestinal epithelial cells

To gain insight on the biological effects of the exocellular factors produced by Bacillus cereus, culture filtrate supernatants of different strains were coincubated with differentiated Caco-2 cells. Exocellular factors were able to detach enterocyte-like cells from the substratum after 1 h of incubation. In addition, microvilli effacing and dramatic changes on the cellular surface of enterocytes were found after incubation periods as short as 20 min. Since cell detachment was not inhibited by fetal calf serum, thiol activated cholesterol-binding cytolysin, cereolysin O, does not seem to be involved. Also, translocation of phosphatidylserine from the inner to the outer leaflets of the plasma membrane was demonstrated by using fluorescein isothiocyanate (FITC)-Annexin V. In contrast to the high capability of detaching Caco-2 cells shown by all the strains under study, the mitochondrial dehydrogenase activity was lowered by culture filtrate supernatants in a strain-dependent manner. For strain M2, the decrease in dehydrogenase activity was already evident after 30 min of incubation. Production of biologically active factors depends on the growth phase, and maximal activity was found in late exponential-early stationary phases. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of concentrated exocellular factors showed a very complex scenery supporting the multifactorial character of the biological activity of B. cereus.     

Rapid effects of pesticides on human granulosa-lutein cells

Following our previous demonstration that p,p'-DDE (dichlorodiphenylchloroethylene), at environmentally relevant concentrations, can rapidly increase intracellular calcium [Ca2+]i concentrations in human granulosa-lutein cells, we examined whether other pesticides, such as Kepone, o,p-DDE and methoxychlor, have similar effects. Cultured human granulosa-lutein cells were loaded with Fura-2 AM, and changes in [Ca2+]i concentrations within small areas of single cells were studied with a dynamic digital Ca2+ imaging system. Kepone, at concentrations of 0.2-2 nmol/ml, consistently increased [Ca2+]i concentrations 2-6 times higher than baseline values within minutes of exposure. Methoxychlor at concentrations of 2.8-280 nmol/ml failed to alter [Ca2+]i levels consistently in cells from 10 patients. However, at 0.28 and 1.4 nmol/ml, increases in [Ca2+]i concentrations could be elicited by methoxychlor. The isomer o,p-DDE at 3 nmol/ml increased [Ca2+]i in granulosa cells of 11/20 patients. Pertussis toxin treatment inhibited the [Ca2+]i increases induced by estradiol, p,p'-DDE, o,p-DDE and methoxychlor, but not by Kepone or progesterone, indicating that Kepone and progesterone may act through an insensitive G protein-coupled receptor. The [Ca2+]i increases induced by Kepone also occurred in Ca2+-free medium, suggesting that [Ca2+]i mobilization occurred from the smooth endoplasmic reticulum. Thapsigargin and cyclopiazonic acid, two inhibitors of the endoplasmic reticulum Ca2+ pump, also stimulated [Ca2+]i increases but did not inhibit the Ca2+ response to all the pesticides. These results demonstrate that pesticides can have a rapid effect on human granulosa-lutein cells, and a nongenomic mechanism of action is suggested.     

Quality characteristics of yogurts fermented with short-chain fatty acid-producing probiotics and their effects on mucin production and probiotic adhesion onto human colon epithelial cells

Probiotics can ferment nondigestible carbohydrates and produce short-chain fatty acids (SCFA; acetate, propionate, and butyrate) in the human colon. In this study, the levels of SCFA were determined in the following yogurts fermented with different combinations of probiotics: (1) cocultures of Streptococcus thermophilus and Lactobacillus bulgaricus (control, C); (2) S. thermophilus, L. bulgaricus, and Bifidobacterium bifidum (C-Bb); (3) S. thermophilus, L. bulgaricus, and Lactobacillus acidophilus (C-La); and (4) S. thermophilus, L. bulgaricus, and Lactobacillus gasseri (C-Lg). Results showed that the acetate levels were significantly higher in C-Bb, C-La, and C-Lg yogurts than in C yogurt. Fermentation and physicochemical characteristics of all yogurts were identical. Treatment of mucus-secreting colon epithelial cells (HT29-MTX) with C-Bb, C-La, and C-Lg yogurt supernatants resulted in an increase in the expression of MUC2 and CDX2 and the production of mucin proteins. The adhesion of probiotics onto HT29-MTX cells increased following treatment with C-Bb, C-La, and C-Lg yogurt supernatants. Our data suggest that a yogurt diet rich in acetate improves the protective function of the intestinal epithelium.     

人α-乳白蛋白在奶牛乳腺上皮细胞中的表达

将人α-乳白蛋白(α-LA)0.76 kb的cDNA序列连接到PSV载体SV40启动子后,构建真核表达载体hα-LA-psv.将构建的真核表达载体转染奶牛乳腺上皮细胞,Aα-乳白蛋白的表达量约为0.85 g/L.结果表明,构建的真核表达载体能够在体外培养的牛乳腺上皮细胞中高效表达Aα-乳白蛋白.     

Synergistic effect of D-glucose and epidermal growth factor display on dynamic behaviors of human epithelial cells

We investigated the synergistic effect of D-glucose and epidermal growth factor (EGF) display on the dynamic cellular behaviors of morphology and migration in a culture of human epithelial cells. The time-lapse observation revealed that the cells on the D-glucose/EGF-displayed substrate were endowed with enhanced migration, accompanied with periodic changes in morphology between round and stretched shapes. Immunofluorescence staining of phosphotyrosine PY20 and vinculin was conducted to determine the intracellular localization of phosphorylated tyrosine expression and focal contact formation, respectively. On the substrate displaying D-glucose and EGF, the cells exhibited increases in the levels of the expression of phosphorylated tyrosine and the formation of focal contacts not only at the cellular periphery but also in the cell body. These findings supported the consideration that the displayed D-glucose causes the cells to be in close contact with the surface via grasping glucose transporters on the cytoplasmic membrane.     

Differential effects of retinoids on proliferation of bovine mammary epithelial cells in collagen gel culture

The effects of increasing concentrations of retinol, retinal and retinoic acid on proliferation of bovine mammary epithelial cells were investigated in collagen gel cultures. All retinoids significantly inhibited proliferation of mammary epithelial cells. The relative inhibitory potency of the retinoids was: retinoic acid > retinal > retinol. Maximal inhibition at 10 microg/ml corresponded to a 75-95% inhibition of proliferation obtained in basal medium. Retinol, retinal and retinoic acid also inhibited proliferation of cells growth-stimulated with insulin-like growth factor-I (IGF-I). Retinoids in highest concentrations (10 microg/ml) inhibited 68-85% of proliferation of cells obtained in culture medium containing 25 ng IGF-I/ml. Retinol and retinoic acid also inhibited proliferation of cells growth-stimulated by insulin and other growth factors from the IGF growth factor family (des(1-3)IGF-I and IGF-II), as well as growth factors from the epidermal growth factor family (EGF and TGF-alpha), with retinoic acid being more effective than retinol. At a concentration of 100 ng/ml retinol and retinoic acid inhibited respectively 24-38 and 44-52% of mammary cell proliferation stimulated by growth factors of the IGF family, and at 10,000 ng/ml, 61-71% of cell proliferation was inhibited. The growth-stimulating effect of insulin, EGF and TGF-alpha was inhibited 42-64% by retinol and retinoic acid at 100 ng/ml, and 64-84% at 10,000 ng/ml. The present results show that retinol, retinal and retinoic acid are potent inhibitors of bovine mammary epithelial cell proliferation. It is suggested that retinoids may have concentration-dependent roles in regulation of pubertal mammary growth and development, indicating that the milk yield potential of heifers may be affected by vitamin A status.