Browsing by Author "Mohamed, Aly Fahmy"

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THE BACILLUS CALMETTE-GUéRIN DERIVED PURIFIED PROTEIN (PPD) POTENTIATES IN-VITRO ANTI-CANCER ACTIVITY OF CERASTES CERASTES SNAKE VENOM IN COLON AND PROSTATE CANCER CELLS
(Inventi, 2017-06) Sabatier, Jean-Marc; Shebl, Rania I; Bakkar, Ashraf; Mohamed, Aly Fahmy; Ayman, Mohamed
Prostate and colon cancer represent a major health problem worldwide. In the present study, we evaluated the anticancer properties and cytotoxicity of Cerastes-cerastes (CC) snake venom on colon (Caco-2) and prostate (PC-3) cancer cells after their pretreatment with variable concentrations of Bacillus Calmette-GuÃ©rin (BCG) derived purified protein derivative (PPD). We monitoned the cell cycle arrest profile and specific cellular apoptosis markers (i.e. pro- and anti-apoptotic genes P53, Bax and Bcl-2 in CC- and BCG/PPD-pretreated cells using real time PCR. The cytotoxicity was determined by using MTT assay. Our data show that 24 h-treatment of cancer cells with CC venom induced a concentration-dependent cytotoxicity with IC50 values of 60 (Caco-2 cells) and 81 (PC-3 cells) Î¼g/ml. Interestingly, addition of BCG/PPD at 25 and 50 Î¼g/ml markedly increased the CC venom-induced toxicity on cancer cells, with IC50 values of 1.04 and 0.59 Î¼g/ml for Caco-2 (up to 102-fold increase) or 2.78 and 0.70 Î¼g/ml for PC-3 cells (up to 116-fold increase). By analyzing the cell cycle arrest and related gene expression pattern, the main phase of cell cycle arrest was found to be G2/M in both cell lines. An S-phase arrest was also observed in PPD pretreated colon Caco-2 cell line to a greater extent than that observed in cells only treated with CC venom. Up regulation of proapoptotic and down regulation of anti-apoptotic genes in PPD pretreated cells were significantly enhanced as compared to cells treated with CC venom alone. In this study, we suggest that PPD -via its synergistic action with the CC venom-might be used as an enhancer of the anti-cancer properties of CC venom.
EVALUATION OF THE IMMUNOGENICITY OF EACH OF L-AMINO OXIDASE- AND L-ASCORBIC ACID-INACTIVATED HEPATITIS A VIRUS IN MICE AS POTENTIAL VACCINE CANDIDATES
(SLOVAK UNIV AGRICULTURE NITRA, 2016-01) Mohamed, Aly Fahmy; Abdulall, Abeer Khairy; Tawfick, Mahmoud Mohamed); Abdullatif, Amal Osman
Hepatitis A virus (HAV) is one of the most common causes of acute viral hepatitis worldwide. Formaldehyde is the currently used inactivating agent in HAV vaccine processing despite of its adverse effects. The current study aimed to evaluate both L-amino acid oxidase (LAO) and L-ascorbic acid (LAA) as alternative inactivants for HAV and the immunogenicity of inactivated HAV in mice. Vero cell line was used for cultivation of HAV. The cytotoxicity of LAO and LAA on Vero cells was evaluated using 3-(4,5-dimethylthiazol-2-yl) 2,5 diphenyl tetrazolium bromide (MTT) assay. The immunogenicity of each LAO- and LAA-inactivated HAV was examined in parallel with reference HAV vaccine in mice. Humoral (total IgG) and cellular immune responses (IFN-gamma and IL-5) were evaluated in mice sera using ELISA. Both LAO and LAA could efficiently inactivate HAV within 30 and 36 hrs post treatment, respectively, at concentrations of 0.4 mu gm/ml of LAO and 1.5 mg/ml of LAA. Inactivated vaccines were immunogenic to mice on both the humoral and cellular levels. LAO prepared vaccines showed a more promising immune reactivity than LAA prepared ones and alum-adsorbed vaccines were more immunogenic than non-adjuvanted ones. In conclusion, data recorded suggest that both LAO and LAA can be used as inactivating agents for HAV grown in cell culture. LAA- and LAO-inactivated HAV can be potential vaccines as they provide effective humoral and cellular immune responses comparable to that of the reference vaccine. The stability of test vaccines is recommended to be traced at different thermal conditions, in addition to different stabilizers and different pharmaceutical formulations must be tested trying to produce a lyophilized formula for long-term stability.
Grape Seed Extract Induces G2/M Cell Cycle Arrest and Apoptosis Via Generation of Reactive Oxygen Species in Breast and Colon Cancer Cell lines
(Inventi, 2017) Sabatseir, Jean Marc; Shebl, Rania Ibrahim; Bakkar, Ashraf; Mohamed, Aly Fahmy; Yaseen, Reem A
Grape seed as a fruit waste was found to have medicinal potentials, specially its procyanidine containment. The present work aimed to investigate the anticancer potential against two of the mostly spread cancers worldwide namely breast and colon cancers. Data recorded revealed that grape seed extract (GSE) demonstrated a dose dependent cytotoxic potential with an IC50 values of 148.5 and 190.2 Î¼g/ml for CaCo-2 and MCF-7 respectively. Gene expression pattern of apoptosis related genes revealed that GSE stimulates a p53-independent apoptotic pathway which was associated with G2/M phase arrest (pË‚0.05). The elevated level of reactive oxygen species (ROS) in the order of 28.5% and 40.8% in CaCo-2 and MCF-7, respectively suggested that GSE is involved also in initiating the intrinsic pathway of apoptosis. These findings focus on the potential of using GSE as a multi-targeted cancer therapy to overcome the problem of resistance to current treatments.
PREPARATION OF A MORE STABLE LYOPHILIZED BCG-T WITH THE SAME BIOPOTENTIAL FOR BLADDER CANCER IMMUNOTHERAPY
(Inventi, 4/15/2017) Mohamed, Aly Fahmy; Bakkar, Ashraf; Yahia, Mohamed; Yasser, Menatallah
BCG-T is a new trend for immunotherapy of bladder cancer as an alternative to chemotherapy. The available product is liquid form of a limited stability profile. The present study aims to prepare a lyophilized form of BCG-T using different stabilizer namely Na glutamate and lactose. Stability of pharmaceutical lyophilized injection dosage form compared with the liquid form was assessed revealing that the Na glutamate and lactose stabilized BCG-T was significantly more stable than BCG-T in the liquid formula. Also, the dry weight and relative humidity and safety of both Na glutamate and lactose stabilized BCG-T were insignificantly different from the liquid form (P >0.05).
ZINC OXIDE NANORODS INDUCED APOPTOSIS IN HUMAN PROSTATIC AND HEPATOCELLULAR CARCINOMA VIA MITOCHONDRIA DYSFUNCTION MEDIATED THOUGH BAX/ BCL-2 WITH P53 ACTIVATION
(Inventi, 2017-06) Sabatier, Jean–Marc; Bakkar, Ashraf; Shebl, Rania I; Mohamed, Aly Fahmy; Hassan, Amr; Gamal, Omar
The present study aimed to experimentally synthesis zinc oxide nanorods (ZnO NRs) using albumin as bio-template by a sol-gel method and to characterize the products using UV-Visible, FTIR, XRD, TGA and HRTEM. The crystallinity and morphology of the ZnO NRs were confirmed to have an average diameter of 70 nm and 250 nm length. The formation mechanism depends on the nucleation of Zn+2 in sites of the albumin followed by Zn+2 assembly in the cavity of albumin and finally thermal treatment to form ZnO in rod shape then calcination to final form ZnO NRs form as shown in HRTEM. Cytotoxicity of developed ZnO-NRs was conducted using MTT assay on both HepG2 and PC-3 cells. Data revealed that ZnO-NPs were toxic to both HepG2 and PC-3 cell lines displayed a concentration dependent viability. The flow cytometry illustrated that apoptosis of hepatocellular carcinoma (HepG2) depends on the cell growth arrest at G1/S phase indicated that inhibition Cyclin E (CDK 2) while prostatic carcinoma (PC-3) depends cell growth arrest at G2/M phase indicated that inhibition of Cyclin ACDK1. The apoptotic mechanism was investigated using rt-PCR. The apoptotic mechanism in both cell lines HepG2 and PC-3 was depended on the upregulated of Bax protein and down regulation of Bcl-2 indicated that the mechanism of mitochondrial outer membrane permeability (MOMP) or mitochondria dysfunction was dependent on activation of P53 protein.