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  • ISSN: 2333-7117
    Early Online
    Volume 3, Issue 5
    Short Communication
    Ipsita Chakravarty, Kanika Kundu, Sumedha Ojha and Subir Kundu
    Bioreactor design is an integral part of bioprocess intensification. The sole aim of process design is to identify the thrust areas and improve the production of the desired metabolite. Complex secondary metabolites like antibiotics are of great importance. Several researchers have highlighted the significance of media formulation, feed strategies, and control of physico-chemical parameters, metabolic engineering and genetic improvement for enhanced antibiotic fermentation. But, there is a lacuna regarding various aspects of bioreactor design and analysis for antibiotic fermentation. In this short communication, we have summarized our research work/ findings on improved antibiotic production through strategic design and development of bioreactors as a case study. The physical design and operational conditions can significantly tackle problems of substrate inhibition, poor cell growth and improper oxygen mass transfer. Thus, helping in meliorated production of several life-saving drugs.
    Kim Marquart, Patrick Schuler, Martin Gamer, Patrick Schulz, Kerstin Otte, Harald Bradl, Ingo Gorr and Simon Fischer*
    MicroRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally control mRNA abundance and thus critically regulate cellular phenotypes. Inter alia, efforts were made to engineer Chinese hamster ovary (CHO) cells which represent the most frequently applied host cell type for commercial production of therapeutic proteins. In the present study, we established a high-throughput screening approach which enabled us to assess the power of miRNAs in CHO cell line engineering approaches. To this end, a robust cultivation method was developed mimicking appropriate cultivation conditions for CHO manufacturing cell lines in micro-scale format. Different chemical transfection reagents were tested for their capability to efficiently introduce small RNAs such as siRNAs and miRNA mimics into CHO cells. We found that the novel cationic lipid Screen FectA plus exhibits excellent transfection efficiency and functionality in various CHO cell lines cultivated in complex production media. Recent approaches using miRNAs in CHO cells were limited to model cell lines that were far away from industry standard or produced standard IgG molecules. The experiments described here were performed using industrial production CHO cell lines stably expressing different monoclonal antibodies as well as novel antibody-derived molecules at various expression levels. This study demonstrates that the identification of a suitable transfection reagent and the careful optimization of the transfection conditions in combination with a micro-scale cultivation process led to the development of a high-throughput miRNA screening platform. A feasibility study using known pro-productive miRNAs rendered this approach suitable for future attempts aiming at the identification of novel miRNA candidates for innovative CHO cell engineering strategies.
    Zhi Dong Zhou*, Yin Xia Chao, and Eng King Tan
    Parkinson's disease (PD) is the second most common neurodegenerative disease worldwide. The mitochondria dysfunction has long been appreciated in PD pathogenesis and can be the common pathological pathway contributing to dopaminergic (DA) neuron degeneration in PD. However mitochondria have its own quality control (QC) defensive systems to protect against stresses induced mitochondria impairment and cell degeneration. The mitochondrial QC system includes cellular QC and molecular QC systems. The mitochondrial cellular QC system includes mitophagy, mitofusion and mitofission processes. The mitochondria molecular QC system mainly refers to mitochondria unfolded protein response (mtUPR), including stress induced activation of expression of nucleus encoded mitochondria chaperones and proteases, which can be sent back to mitochondria to counteract against stress induced mitochondria impairment. So far little is known about mtUPR signaling pathway and its relevance to human diseases. The disturbance of mtUPR can be the cause for genetic factors as well as environmental factors induced mitochondria impairment and DA neuron degeneration in PD. The screening and identification of novel small molecular weight neuroprotective mtUPR activators can help develop future anti-PD drugs to alleviate progressive DA neuron degeneration in PD. Furthermore novel molecular targets identified in mtUPR signaling pathway and the potential crosstalk between mtUPR and PD relevant genes can add to PD pathogenesis and therapy. In this short review, these issues will be analyzed and discussed.
    Else Driehuis and Hans Clevers*
    TMPRSS2-ERG fusions are common genetic events in prostate cancer. Until now, this genetic alteration was modelled by ERG overexpression. In this short communication, we report the creation of mouse prostate organoids that have undergone gene fusion through a CRISPR/Cas9-based strategy. The genetic fusion of TMPRSS2 and ERG results in ERG overexpression. This effect is androgen receptor-mediated, as expression of the fusion transcript can be restored to wildtype ERG levels by treatment with the androgen receptor antagonist Nilutamide.
    Dennise Lizarraga-Lizarraga, Ataulfo Martinez-Torres, Raul Llera-Herrera, Angeles Edith Espino-Saldana, and Alejandra Garcia-Gasca*
    Myostatin, a negative regulator of skeletal muscle growth, is also expressed in cardiac muscle; however its function in this tissue is poorly understood. In this study, the CRISPR/Cas9 system was used to inactivate mstn in zebrafish. Microinjection was performed in one-cell stage embryos, and genomic DNA sequencing analysis confirmed high mutation efficiency. Coronal histological sections from 5 days post-fertilization (dpf) larvae showed the expected phenotype of skeletal muscle growth, but no evident morphological alterations were observed in the heart. An examination of 2 dpf embryos revealed a significant decrease in the heart rate, suggesting that mstn could play a role in cardiac function during development.
    Review Article
    Guochao Li and Yingli Sun*
    As an important part of precision medicine, liquid biopsy attracts more and more attention of scholars and clinicians. It refers to the real-time monitoring of the dynamic alterations of tumor by detecting circulating tumor cells (CTCs), circulating tumor DNA (ctDNA) and exosomes in patients plasma or serum. Thus, liquid biopsy has irreplaceable advantages in tumor early diagnosis, progression monitoring, curative effects evaluation, prognosis judgement and so on as a non-invasive method. Here we reviewed traditional clinical diagnostic technologies and the liquid biopsy and summarized the advantages, limitations and test indexes of CTCs, ctDNAs and exosomes for their clinical applications. We also enumerated other applications of liquid biopsy by using other body fluids to monitor some particular cancers. At last, we forecasted the opportunities and existed challenges of liquid biopsy.
    Jixin Ding, M. Reza Saadatzadeh, Aaron Cohen-Gadol, and Karen E. Pollok*
    Development of effective anti-cancer therapies continues to be a challenge due to genetic instability that promotes intra-tumoral molecular heterogeneity and subsequent adaptation to therapy. In this review, we briefly discuss the history of the p53 family (p53/p63/p73) and how regulatory circuits affect its function in promoting apoptotic cell death. In addition, we provide perspective on how protein-protein interactions between members of the p53 family, as well as, with other regulatory proteins can dictate response to therapy. In the future, therapies that focus on targeting the p53 family to sustain pro-apoptotic pathways in combination with cancer-specific dysregulated signaling pathways is a promising approach. With further investigation at the basic science level, improvements in efficacy and quality of life for cancer patients with diverse molecular signatures may be realized.
    Research Article
    Gopalan Rajkumar, Periyakali Saravana Bhavan*, Veeran Srinivasan, Annamalai Asaikutti, and Rajendran Udayasuriyan
    The in vitro and in vivo antibacterial activity of marine alga, Turbinaria ornata against Pseudomonas aeruginosa on Macrobrachium rosenbergii was studied. The hexanic, acetonic and methanolic extracts of T. ornata were subjected to in vitro study, which showed that the methanolic extract of T. ornata was worked well and produced 24.02 0.47mm zone of inhibition against P. aeruginosa. In the in vivo study, the characteristic appearance of brown/black spots was detected all over the body of M. rosenbergii when the prawns were subjected to immersion in water containing P. aeruginosa (107 cells/mL) and fed with basal diet contained no incorporated T. ornata. Activities of metabolic enzymes (GOT and GPT) and antioxidant enzymes (SOD and catalase), and lipid peroxidation (LPO) were found to be significantly increased in the hepatopancreas of M. rosenbergii due to the infection with P. aeruginosa. These effects were greatly reduced/ neutralized when M. rosenbergii fed with methanolic extract of T. ornata incorporated feed, which in turn ultimately enhanced the survival rate of M. rosenbergii against P. aeruginosa infection. The 2D gel electrophoresis revealed degradation of hepatopancreatic proteins in P. aeruginosa infected M. rosenbergii, whereas the reverse was seen in methanolic extracts of T. ornata incorporated feed fed prawns. Thus, T. ornata possessed the potency of anti P. aeruginosa. One of the active principles of T. ornata, 2-Hexadecen-1-ol, 3,7,11,15-tetramethyl-,[R-[R*,R*-(E)]] was found to be bind with the protein, exotoxin-A of P. aeruginosa when molecular docking was performed. Therefore, there is scope for developing aquaculture medicine with T. ornata against Pseudomonas infection.
    Special Issue on Diverse Roles of Nitric Oxide in Biomaterials and Implants
    Review Article
    Petukhov VI1*, Baumane LH2, Dmitriev EV3 and Vanin АF4
    Abstract: The paper informs about the specific shifts in metal-ligand homeostasis (MLH) of epidermic cells (hair) in Chernobyl accident liquidators, which are viewed by the authors as biomarkers of oxidative/nitrosative stress. The detected (by EPR-analysis) relationships between NO-production and MLH quantitative shifts can be indicative of the possible participation of nitric oxide in generation of cell electric potential.
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