Effects of Major Ingredients in Cattle Milk on Enzyme Kinetics of Recombinant β-galactosidase (BGalP) Expressed in Pichia pastoris

Background and objective: β-galactosidase enzymes hydrolyze lactose into glucose and galactose for production of lactose free dairy products. However, different ions and fat content in milk may act as the inhibitor or activator for β-galactosidase enzymes. A cold-active β-galactosidase enzyme (BGalP), originally from Planococcus sp. L4, was previously expressed in Pichia pastoris to perform lactose hydrolysis in the refrigerated milk. In this study, the effects of milks major ingredients were evaluated on the enzymatic kinetics to confirm its capacity for hydrolyzing milk lactose.Material and methods: The activity was determined in different concentrations of NaCl, KCl, MgCl2, and CaCl2 as well as in the milk with low, medium or high-fat content. In these experiments ortho-Nitrophenyl β-galactoside was used as the substrate. Additionally, glucose was measured as the product after incubation of milk with BGalP enzyme for 24 h at room temperature.Results and conclusion: This study demonstrated that ions and fat content did not adversely affect the enzyme activity in the concentration corresponding to the milk contents. Ca (27.5-32.5 mM), Cl (25.3-30.9 mM), Na (15.2-39.1 mM) and Mg (3.75-5.83 mM) had no inhibitory effects, but KCl decreased the enzyme activity. Since Cl existed in MgCl2, and CaCl2 exerted no inhibitory effects, it can be concluded that inhibitory effects of KCl resulted from potassium rather than chloride. The results indicate that BGalP enzyme was not inhibited by milks major ingredients and has the potential to be used for the production of lactose-free dairy products.Conflict of interest: The authors declare no conflict of interest

Effects of Zataria multiflora Extract and Carvacrol on Doxorubicin-Induced Oxidative Stress in Rat Brain

Background: Due to the antioxidant effects of Zataria multiflora (ZM) and Carvacrol (CAR) in various problems and the prominent role of the ROS in neurotoxicity induced by Doxorubicin (DOX), this study was designed to investigate the effects of ZM hydroalcoholic extract and CAR on DOX-induced oxidative stress in rat brain. Methods: 24 male rats were randomly divided into four groups including: 1)Control ,2)Doxorubicin (DOX) that received DOX via a tail vein on the first day of the study, 3,4) ZM+DOX and CAR+DOX which received ZM and CAR by gavage for 28 consecutive days. Brain tissue removed for redox markers evaluation. Results: MDA level in the DOX group was significantly increased compared to control group while in treated groups did not show any significant changes in comparison with the DOX group. Also, Thiol content in DOX group showed significant reduction compared to control group. Thiol contents in treated groups showed no significant difference compared to DOX group. Catalase (CAT) activity, an antioxidant enzyme, in the DOX group were significantly decreased compared to control group and increased in treated rats in comparison with the DOX group. Activity of Superoxide dismutase (SOD), an antioxidant enzyme, in the DOX group was significantly reduced compared to control group and increased in treated rats in comparison with the DOX group. Conclusion: The present study showed that ZM hydroalcoholic extract and CAR could inhibit DOX induced oxidative stress of the brain mainly with effect on the enzymatic antioxidant defense system

Effects of standardized Zataria multiflora extract and its major ingredient, Carvacrol, on Adriamycin-induced hepatotoxicity in ratAt a glance commentary

Background: At a glance commentaryScientific background on the subject Since Adriamycin is used as an anticancer drug among various communities and is one of its side-effects, it is important to find a way to prevent this complication. Of these, medicinal plants such as Zataria multiflora can be important in preventing the complications.What this study adds to the field This study shows that carvacrol has a stronger effect on the liver antioxidant system as well as hepatic enzymes activity than the whole plant extract, and then carvacrol can be more helpful than whole plant in the treatment of hepatic damage caused by Adriamycin.Due to antioxidant effects of Zataria multiflora (ZM) and Carvacrol (CAR) in many cases and the prominent role of reactive oxygen species (ROS) in hepatotoxicity induced by Adriamycin (ADR), the aim of this study is to investigate the effects of ZM and CAR on ADR-induced hepatotoxicity. Methods: Twenty four male Wistar rats were randomly divided into four groups including: 1)Control, 2)Adriamycin (ADR), 3,4) ZM + ADR and CAR + ADR that received ZM and CAR for 28 consecutive days. Blood samples were collected on the days 0, 14 and 28 to determine the alkaline phosphatase (ALP), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels. Also, the hepatic redox markers were evaluated. Results: ADR significantly increased ALP, ALT and AST in comparison with the control (p < 0.05 - p < 0.001). In CAR + ADR group, the serum ALP, ALT and AST were significantly reduced compared to those of the ADR group (p < 0.01 to p < 0.001). Also, in ZM + ADR group, serum ALP and ALT compared to ADR was significantly reduced (p < 0.001). MDA level in the ADR group significantly increased compared to control (p < 0.01). The MDA level in ZM + ADR (p < 0.05) and CAR + ADR (p < 0.01) groups were significantly reduced compared to that of ADR. Thiol levels in ZM + ADR group significantly increased compared to the ADR group (p < 0.05). The activities of CAT in the ADR group was significantly reduced compared to control (p < 0.05) and increased in treatment groups in comparison with the ADR (p < 0.01). Conclusion: Long-term administration of ZM extract and CAR could reduce the oxidative damage in the rat liver induced by ADR through the strengthening of the antioxidant system. Keywords: Adriamycin, Zataria multiflora, Carvacrol, Hepatotoxicit

Hydroalcoholic extract of psoralea drupacea inhibits proliferation and migration of hepatocellular carcinoma cells and decreases angiogenesis in chick chorioallantoic membrane

Objective: Hepatocellular carcinoma is one of the leading causes of cancer-related death worldwide. Experimental studies reported that some plants in the genus of Psoralea (Fabaceae family) show anticancer potential. The present study aimed to evaluate the effects of Psoralea drupacea extract (PDE) on HepG2 liver cancer cells. Methods: The proliferation, cell cycle, and migration of HepG2 cells were determined by thiazolyl blue tetrazolium bromide test, propidium iodide staining, and scratch assay, respectively. The effects of PDE on the activity of matrix metalloproteinases (MMPs) and angiogenesis were evaluated by the gelatin zymography method and chicken chorioallantoic membrane model, respectively. Results: The culture of HepG2 cells in the presence of PDE (24 hr and 48 hr) significantly reduced their viability (at a concentration of ≥ 50 µg/mL) and increased the percentage of cells in the sub-G1 stage. PDE also increased the antiproliferative and proapoptotic activities of doxorubicin (3 and 6 µg/mL). The extract significantly decreased the generation of reactive oxygen species and lipid peroxidation in the cells. Moreover, PDE (25 and 50 µg/mL) significantly suppressed the migration ability of HepG2 cells, which was associated with inhibition in the activity of MMP2 and MMP9 (50 µg/mL). Furthermore, treatment with PDE significantly reduced the number and diameter of vessels in the chick chorioallantoic membrane. Conclusion: PDE decreased the survival and cell cycle progression of liver cancer cells through a mechanism other than oxidative stress. This extract also showed an anti-angiogenesis effect and diminished the migration ability of HepG2 cells by inhibiting MMP activit

Anticancer Mechanisms of Berberine: A Good Choice for Glioblasto-ma Multiforme Therapy

The most typical malignant brain tumor, glioblastoma multiforme (GBM), seems to have a grim outcome, despite the intensive multi-modality interventions. Literature suggests that biologically active phytomolecules may exert anticancer properties by regulating several signaling pathways. Berberine, an isoquinoline alkaloid, has various pharmacological applications to combat severe diseases like cancer. Mechanistically, it inhibits cell proliferation and invasion, suppresses tumor angiogenesis, and induces cell apoptosis. The antitumoral effect of berberine in GBM is increasingly recognized. This review sheds new light on the regulatory signaling mechanisms of berberine in various cancers, proposing its potential role as a therapeutic agent for GBM

Auraptene-induced cytotoxicity mechanisms in human malignant glioblastoma (U87) cells

Glioblastoma multiforme (GBM), like the devastating type of astrocytic tumors, is one of the most challenging cancers to treat owing to its aggressive nature. Auraptene, as a prenyloxy coumarin from citrus species, represents antioxidant and antitumor activities; however, the underlying antitumor mechanisms of auraptene against GBM remain unclear. The present study aimed to evaluate the cytotoxic and apoptogenic effect s of auraptene, as a promising natural product, and the possible signaling pathways affected in human malignant GBM (U87) cells. Reactive oxygen species (ROS) production significantly decreased in the first 2, and 6 hours after treatment with auraptene however, ROS levels increased in other incubation times (8 and 24 hours), dramatically. N-acetyl-cysteine (NAC) markedly attenuated auraptene–induced ROS production, and consequently reversed auraptene–induced cytotoxicity in 8 and 24 hours after treatment, as well. Induction of apoptosis occurred in the first 24- and 48-hours concentration-dependently. The qRT-PCR showed an up-regulation in p21, CXCL3, and a down-regulation in Cyclin D1 genes expression. Western blot analysis confirmed the up-regulation of the Bax/Bcl-2 ratio protein levels concentration-dependently. Hence, this study collectively revealed that the increase in ROS level is at least one of the mechanisms associated with auraptene-induced GBM cell toxicity as well as the induction of apoptosis through Bax/Bcl-2 modulation and genes expression involved that contribute to the cytotoxicity of auraptene in U87 cells. So, auraptene might be utilized as a potential novel anti-GBM agent after further studies