While not reported at the time, it is now understood that injury mechanisms in electrical shock trauma include not only thermal damage produced by Joule heating but also in superposition tissue electroporation and macromolecule degeneration (Lee et al., 2000). [44] Evidence for the existence of such pre-pores comes mostly from the "flickering" of pores, which suggests a transition between conductive and insulating states. Artificially induced cell fusion can be used to investigate and treat different diseases, like diabetes,[7][8][9] regenerate axons of the central nerve system,[10] and produce cells with desired properties, such as in cell vaccines for cancer immunotherapy. It is generally accepted that for a given pulse duration and shape, a specific transmembrane voltage threshold exists for the manifestation of the electroporation phenomenon (from 0.5 V to 1 V). Numerical techniques are then discussed for simulating treatments based on derivations from Maxwell's equations. (B) Applying an electric field drives the negatively charged DNA strands away from the cathode and toward the anode. About the organisms, guns are used mostly for plant-cells and few other "hard to electroporate" organisms. Study concluded, that gene electrotransfer with pIL-12 is safe and well tolerated. The general method of transformation is the chemical transformation in which the treatment of host cells with calcium-chloride makes the cells more permeable to take up exogenous DNA. However, some subjects experienced strong muscle contractions occurring with each electrical pulse, and around 25% suffered mild muscle fatigue after treatment. Yang et al. Further, by tuning the electrical pulse parameters, electroporation can induce cell death without any significant levels of accompanying thermal damage (Davalos et al., 2005), giving rise to many of the benefits seen clinically, which are discussed later in this chapter. Though ATMT method is effective, it is time consuming and hinders further application in genetic engineering. [11] However, the first and most known application of cell fusion is production of monoclonal antibodies in hybridoma technology, where hybrid cell lines (hybridomas) are formed by fusing specific antibody-producing B lymphocytes with a myeloma (B lymphocyte cancer) cell line.[12]. This membrane permeability increase is used for many applications in biotechnology, medicine and the food industry. In utero electroporation surgeries begin by exposing the embryonic pups of a pregnant animal, usually a rodent. Using a gene gun directly shoots a piece of DNA into the recipient plant tissue. Proteins, the extracellular matrix, and critical structures such as blood vessels and nerves are all unaffected and left healthy by this treatment. Based on established marine Nannochloropsis species electrotransformation protocol, nuclear genetic transformation was established in N. limnetica, meanwhile the appropriate antibiotic selection concentration and electric field strength of electroporation were determined. The irreversible damage of cell membrane occurs when high-intensity PEF treatment is applied. P.T. Direct DNA uptake is useful for both stable transformation and transient gene expression. Method # 4. [23] Irreversible electroporation (IRE) has since proven effective in treating human cancer, with surgeons at Johns Hopkins and other institutions now using the technology to treat pancreatic cancer previously thought to be unresectable.[24]. Reversible electroporation is a precursor to the development of irreversible membrane defects. (1991). [21] Soon also gene electrotransfer became of special interest because of its low cost, easiness of realization and safety. [49] There are a number of factors that can influence the efficiency of gene electrotransfer, such as: temperature, parameters of electric pulses, DNA concentration, electroporation buffer used, cell size and the ability of cells to express transfected genes. The material alteration is thus chemical in nature. This type of procedure has the same tumor ablation success as N-TIRE. Upon application of this potential the membrane charges like a capacitor through the migration of ions from the surrounding solution. There are currently several ongoing clinical studies of gene electrotransfer,[27] where safety, tolerability and effectiveness of immunization with DNA vaccine, which is administered by the electric pulses is monitored. The effectiveness of microalgal electroporation was first reported by Brown et al. (2002) studied the effects of iontophoresis and electroporation on the transdermal delivery of buprenorphine from solutions and hydrogels. The method is used to deliver large variety of therapeutic genes for potential treatment of several diseases, such as: disorders in immune system, tumors, metabolic disorders, monogenetic diseases, cardiovascular diseases, analgesia….[16][17][18]. That is, only the cells within areas where E≧Eth are electroporated. Currently, the most accepted theoretical description of the events underlying electroporation is the aqueous pore formation theory (Weaver and Chizmadzhev, 1996). These bursts of electricity increase the resting transmembrane potential (TMP), so that nanopores form in the plasma membrane. Often, a CT machine will be used to help with the placement of electrodes during the procedure, particularly when the electrodes are being used to treat tumors in the brain.[31]. salt content). Recently, citrus molecular genetics and biotechnology work have been accelerated in the world. 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