Halal issues in the mammalian cell culture for recombinant protein production

Awareness and interest in halal products has significantly increased among muslim and non-muslim countries. Biopharmaceutical products including recombinant therapeutic proteins contribute to a considerable percentage in the worth of the overall global halal industry, which is said to be US$ 2.3 trillion. The number and demand for approved biopharmaceutical products produced from mammalian cell culture methods such as vaccines, monoclonal antibodies, antibodies, hormones, and other therapeutic proteins are increasing worldwide. The mammalian cell culture technology has been known to be perfectly suited to the production of recombinant therapeutic proteins. Mammalian cell culture is a general term used for the isolation of the cells of a mammalian from specific tissues further cultured and reproduced in an artificial growth media. The overall process in the recombinant protein synthesis using mammalian cell culture involves the use ingredients or materials that may be questioned from the perspective of halal procedures that make the product cannot fulfill the requirements of halal pharmaceuticals. This review will discuss the application of mammalian cell culture bioprocesses in recombinant biopharmaceutical protein production with focus on the halal-compliance status of the materials and methods

Dental bio-aerosol: a silent threat in dental practice

Many dental procedures produce extensive bio-aerosols and splatter that are routinely contaminated with bacteria, viruses, and blood. Dental bio-aerosol has been considered as a potential occupational hazard in dental setting since the aerosolized microorganism from a patient’s mouth during dental treatment procedures may stay airborne for long periods of time and inhaled into the lungs of dental health care workers (DHCW). Reducing the risk of exposure to dental bio-aerosol has become a challenge for DHCW. To date, various infection control reports and procedures have been published to inform and educate dental health care personnel about the importance of practicing adequate infection control. This brief review highlights the potential risks that can be encountered with dental bio-aerosol. Various methods undertaken to control the infection caused by the aerosol are also discussed

Site-directed mutagenesis for the production of mutant TP53 gene and analysis of its tumor suppressor activity

The p53 therapy based approach is a good candidate for treatment of TP53-defect cancer. However, this treatment is unsuitable for some cancer cases especially for that caused by dominant-negative activity of the mutant p53 protein. Thus, more effective treatment is needed to overcome this problem and DNA vaccination may be the suitable candidate where mutant p53 may become a suitable candidate as an antigen for the DNA vaccination strategy. Therefore, the research aims to produce mutant TP53R248Q through PCR site–directed mutagenesis and to confirm its tumor suppression ability. The mutation of R248Q was generated via Polymerase Chain Reaction (PCR) site-directed mutagenesis to pCMVp53 plasmid. Following that, these constructed TP53R248Q was transfected into the TP53-null H1299 cell lines in order to investigate its tumor suppression ability, by studying its phenotype and genotype expression. Phenotype study was conducted by using colony formation assay while quantification of TP53 downstream target gene, p21waf1 was conducted for the genotype study. The transfection with exogenous TP53R248Q resulted in massive colony proliferation and downregulation of p21waf1, thus confirmed that the generated mutant TP53 has lost its ability to restrain cells growth. These data therefore confirmed that the PCR site-directed mutagenesis technique has been successfully carried out to induce the desired mutation in the TP53 gene. Thus, this technique may become an interesting option to generate novel recombinant proteins, especially in the development of specifically designed DNA vaccine as a gene therapy in the future

In silico analysis of molecular interactions between the anti- apoptotic protein survivin and dentatin, nordentatin, and quercetin

Survivin is a member of the inhibitor of apoptosis (IAP) family and is reportedly overexpressed in various types of human malignancies. Because the phytochemical compounds dentatin, nordentatin, and quercetin have demonstrated antiproliferative effects in various cancer cell lines, we compared their binding affinities for survivin in silico. Molecular docking analyses were performed using PyMol, Discovery Studio Biovia 2017, AutoDock Vina, and AutoDock Tools version 1.5.4. These computations indicated greater survivin binding affinity of quercetin (ΔG −7.0 kcal/mol) than nordentatin and dentatin (ΔG −6.5 and −5.5 kcal/mol, respectively), but suggest that all three compounds act as ligand inhibitors of survivin. The present data warrant validation using in vitro and in vivo assays

Wound healing properties of biotransformed asiaticoside by aspergillus niger

Biotransformation is extensively used to create useful metabolites from various natural products and as an alternative to chemical synthesis for the preparation of pharmacologically-active compounds. This study investigated the effects of asiaticoside and its biotransformed product on wound healing activities. Asiaticoside, the bioactive constituent of Centella asiatica has been reported to possess wound healing properties. Microbial transformation of asiaticoside using Aspergillus niger was carried out to produce an asiaticoside biotransformed product and the wound healing activities of asiaticoside and its biotransformed product were investigated. Their effects on transforming growth factor-beta 1 (TGFβ1) and tissue inhibitor of metalloproteinase 1 (TIMP1) gene expression were examined to understand the mode of action and on the cell proliferation and wound healing using human keratinocytes. Results of in vitro study showed that asiaticoside concentrations between 7.5 and 120 μg/mL gave higher value of cell proliferation than the negative control. On the other hand, the biotransformed product concentrations between 0.058 and 3.75 μg/mL exhibited high cell viability but the viability was lowest at 15.0 μg/mL, suggesting cytotoxic effects on the cells. In wound healing assays, there were significant differences on wound closure in comparison to the negative control (P<0.05). Both asiaticoside and the biotransformed product increased the expression of TGFβ1 and TIMP1 respectively, with the latter showing more enhanced expressions of both genes. The biotransformed product also showed faster migration and healing rate under microscopic observation

Anti-proliferative bacteriocins active against MRSA from coagulase negative Ent. mundtii strain C4L10 isolated from non-broiler chicken

Bacteriocins produced by a variety of lactic acid bacteria (LAB) are gaining more attention for not only as alternative food preservative but also as therapeutics agent. A bacteriocin, Ent C4L10, was produced by coagulase negative Enterococcus mundtii strain C4L10 (Accession No. KC731423) previously isolated probiotic from Malaysian non-broiler chicken. Based on agar diffusion assay, it showed antimicrobial activities against Methicillin resistance Staphylococcus aureus (MRSA) used as an indicator organism. Approximately 10 kDa protein was purified employing three-phase partitioning (TPP) method and it was shown to be highly thermostable, retaining activities at 121°C for 15min, and was stable in a pH range of 4-9. There was however a loss in activity after protease treatment. PCR amplification using enterocin gene primers showed that Ent C4L10 sequence is highly similar to Bacteriocin L-1077 (83% identity). In order to study its anti-proliferative potential, purified Ent C4L10 was also tested against four human cell lines; i.e., lung cancer (H1299), breast cancer (MCF 7), colon cancer (HCT116) and oral cancer (HSC3). It was found that oral cancer cell line was the most sensitive with a cytotoxic index of IC50 of 9.009µg/ml, followed by breast cancer (IC50 (11.51µg/ml), and the least sensitive was with colon cancer cell line (IC50 of 20.57µg/ml). In conclusion, putative 10 KDa Ent C4L10 is a class IIa bacteriocin isolated from coagulase negative Enterococcus mundtii strain C4L10 shown to have anti-proliferative properties. Therefore, this bacteriocin has not only great potential for use in food preservation, it future use as an antitumor agent should also be explore

Thymoquinone and its pharmacological perspective: A review

Thymoquinone (TQ) a plant-derived dietary of Nigella sativa, is a well-known traditionally and clinically used natural medicine. The diverse pharmacological properties of TQ have been identified including antimicrobial, antihistamine, antioxidant effects, immunomodulator, and anticancer activities. In this review, investigations regarding the effects of TQ in oxidative stress, immunomodulation, and various type of cancer have been reviewed based on the available relevant literature. TQ-based induction of the immune system by modulating different inflammatory mediators i.e., cytokines, leukins, interleukins, interferons, and other immune cells have been reviewed here. Several studies signified remarkable anticancer potentials of TQ depending upon its concentration and type of cancer cell. Conclusively, understanding pharmacological activities of TQ its molecular mechanism could help researchers to develop a potent analog of well-established chemotherapeutic drugs in clinical trials

Antiproliferative activity of Curcusone B from Jatropha Curcas on human cancer cell lines

This study was designed to determine antiproliferative properties of curcusone B from the stem bark of Jatropha curcas on cell lines K562 (chronic myelogeneous leukemia), and H1299 (human non-small cell lung carcinoma). Cells were cultured in the presence of curcusone B at various concentrations and the percentage of cell viability was evaluated by trypan blue exclusion method and MTT assay. Curcusone B showed significant dose-dependent inhibition of cell proliferation of K562 and H1299 cell lines with an IC50 of 6 μg/mL and 15.0 μg/mL, respectively. The findings suggested that the curcusone B had strong antiproliferation which was dose dependent and may have potential as an anticancer agent

Growth Suppression of non-small lung carcinoma cell H1299 transfected with p53 and p73beta

p53 gene has been hailed to have important roles in maintaining genomic integrity. The critical roles of this tumor suppressor in cell cycle, DNA repair, and opoptosis profoundly contribute to development of cancer. p53 mutations have been found in almost fifty percent of all cancer types, including lung cancers. Hence, the idea of restoring the normal function of p53 gene by exogeous p53 replacement therapy has been discussed and investigated in order to overcome cancer. However, introduction of wild-type p53 protein is unable ot induce apoptosis in all tumor cases, at least in part, due to their resistance to exogenous p53. p73Bhas been regarded as one of the strongest candidate as it is similar to p53 in many aspects: structural homology, transactivation of p53-downstrean genes, and induction of apoptosis. It has been reported that a similar therapeutic strategy can be successfully applied in p73 activation as well. Interestingly, the mutation of this gene has been rarely found in cancer, suggesting different mechanism or regulation pathway of this gene. The aim of this study is to compare the tumor suppressor activity of p53 and p73B, on p53 and p73B constructed in pCMV plasmid was conducted through chemical mediated transfection. Introduction of exogenous expression of p53 significantly suppress colony formation of H1299 cells under G418 selection while p73B could partially suppress the colony formation. Consistently, exogenous p53 and p73B raise expression of p21/Waf gene. These results show overlapping regulation performed by p73B which suggest its application as alternative candidate in gene therapy