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Essay: Gene transfer techniques Ė pros and cons. Gene transfer is a technique to stably and efficiently introduce functional genes (that are usually cloned) into the target cells. Genetic transfer is the mechanism by which DNA is transferred from a donor to a recipient. Once donor DNA is inside the recipient, crossing over can occur. The result is a recombinant cell that has a genome different from either the donor or the recipient. A recombination event must occur after transfer in order that the change in the genome be heritable (passed on to the next generation).The genes are the blueprints essential to generate all the proteins in our bodies which eventually perform all the biological functions. Therefore, when a gene is efficiently introduced into a target cell of the host, the protein which is term about encoded by that gene is produced . Gene transfer technologies developed initially as a research tool for essay writing in english sports and games, studying the term paper, gene expression and thesis mempengaruhi its function. A range of techniques and naturally occurring processes are utilized for the gene transfer. Techniques are available for term paper housekeeping, the introduction of DNA into many different cell types in make, culture, either to study gene function and regulation or to produce large amounts of recombinant protein. Cell cultures have therefore been used on a commercial scale to synthesize products such as antibodies, hormones, growth factors, cytokines and viral coat proteins for immunization.The most important application of this technology is in term paper about, vivo gene therapy, i.e. the introduction of DNA into the cells of live animals in order to treat disease.

The extraordinary potential of altered DNA molecule is to give rise to new life-forms that are better adopted for survival. Change in base sequence of DNA leads to change in protein causing disease condition which can be corrected by manipulation of gene. Much emphasis has been given to the gene manipulation in cardiovascular diseases, parkinsonís disease, lysosomal disorders, ocular gene therapy and essay writing osteoarthritis. Overview of gene-transfer strategies: Gene transfer can be achieved essentially via three routes. Paper? The most straightforward is direct DNA transfer, the debra resume, physical introduction of foreign DNA directly into the cell. About? For example, in cultured cells this can be done by microinjection, whereas for cells in vivo direct transfer is often achieved by bombardment with tiny DNA-coated metal particles. The second route is termed transfection, and teaching this encompasses a number of housekeeping, techniques, some chemical and accuplacer essay some physical, which can be used to about persuade cells to essay sports and games take up DNA from term their surroundings.

The third is to package the DNA inside an. animal virus, since viruses have evolved mechanisms to naturally infect cells and introduce their own nucleic acid. The transfer of foreign DNA into a cell by this route is termed transduction. The choice of methods of DNA transfer depends upon the target cells in make me an essay, which transformation will be performed. It also depends upon the objectives of gene manipulation. The transfection may be either stable or transient. Although, choice of about, DNA transfer method is very important, the other important steps are selection of gene, isolation of gene, preparing recombinant DNA and selection of transformed cells. The regeneration of organism with new characteristics is also equally important. The delivery of genes in vitro can be done by treating the cells with viruses such as retrovirus or adenovirus, calcium phosphate, liposomes, particle bombardment, fine needle naked DNA injection, electroporation or any combination of these methods.

These are the powerful tools for research and have possible applications in in english sports and games, gene therapy. FACTORS TO CONSIDER WHEN SELECTING A GENE DELIVERY METHOD. The advancement of new technologies has made gene delivery both more efficient yet more confusing given the formidable array of reagents and methods now available. Determining the best gene delivery approach for any particular experiment requires careful consideration of a number of term paper about housekeeping, different factors as stated below. 1. Cellular Context or Environment Ė The context or environment of the cells (i.e., in who can me an essay for university, vivo vs. in. vitro vs. ex vivo) to which nucleic acids will be delivered is the first factor to consider when. deciding upon a gene transfer method.

The obstacles and challenges posed by delivery to in. vitro cell cultures are very different than those posed by in about, vivo or ex vivo cells, and require. different tools and techniques for overcoming them. For example, targeting nucleic acids to. particular cells while avoiding other cells is usually a major concern for in vivo gene transfer. applications, sometimes a concern for ex vivo applications, but usually not a concern for resume, in vitro. Another important issue that must be addressed when planning in vivo (but not in vitro) gene. delivery studies is the immunogenicity of the term about housekeeping, gene transfer system. This is particularly important when considering the use of certain viral vectors discussed previously, which may be highly immunogenic.

Also related to immunogenicity is the issue of disadvantages of internet writing, general safety, both for the experimenter and for the subject of any study utilizing transfection. Almost always, the use of viral vectors requires very extensive safety precautions than does the use of term, synthetic transfection methods. 2. Cell or Tissue Type Ė The origin of the cells or tissue to which nucleic acids will be. transferred is typically the second parameter considered when selecting a gene delivery. method. For a number of different reasons, the difficulty of essay sports, cellular nucleic acid delivery varies. from cell type to cell type. Because of this, it is always important to review the literature to. determine if the cell type of interest has previously been utilized for gene transfer experiments, which exact methods have been used, and what delivery efficiencies were achieved, i.e., what. percentages of cells actually transfected or transduced, with each method.

Certain cell types are often described as Ďnotoriously difficult to transfectí. These include primary cell cultures of many varieties. Successful gene transfer to such cells. usually requires a good deal of research before selecting any particular gene delivery method. 3. Term? Delivery Efficiency Ė For most applications, transfecting or transducing as many cells as.

possible is preferred, especially when it is make me an essay for university necessary to housekeeping detect the presence of rare transgene. products. Thus, it is important to determine the level of delivery efficiency required for a given. application before deciding on a particular gene transfer method. In most cases, it is mempengaruhi pencapaian found that.

viral vectors provide the highest delivery efficiencies. For some commonly used and easily transfected cell lines, cationic lipids nearly match viral vectors in term paper, their gene transfer efficiency, while providing safer and much quicker procedures. 4. Stable vs. Transient Gene Delivery Ė Gene transfer procedures are frequently categorized. by whether the delivered gene remains separate from the thesis belajar pencapaian, host cell chromosome or whether it is integrated into housekeeping the host cell chromosome. In the first case, known as transient gene delivery, expression of the transgene typically dissipates after a given period of time Ė usually within. several days Ė because the expression vector is either degraded or expelled from the host cell.

In the second case, known as stable gene delivery, expression of the transferred gene is. prolonged, or stable, because the vector is integrated into debra the host cell chromosome. Obviously, different applications require different time periods of transgene expression. Thus, a careful. comparison between different gene delivery methodologies will allow a suitable choice for. generating the desired transient or stable expression. In the case of viral vectors, determining whether a particular virus is suited for stable or transient. transduction is term paper about housekeeping simply a matter of learning the general characteristics of the vector. Debra? In the case.

of chemical or physical/mechanical gene transfer, determining whether a given method is. suitable for transient vs. stable delivery may be as easy as reading sales literature from term paper about housekeeping the. manufacturer or checking primary research literature. 5. Type of Delivered Molecule Ė Another factor to consider when selecting a gene delivery. method is what type of nucleic acid or other molecule is being transfected or transduced. For. example, a reagent that efficiently delivers plasmids may not efficiently deliver DNA. oligonucleotides.

It is frequently easiest to simply review the literature to learn what methods have been used successfully, and to contact other researchers who have experience delivering the molecule of interest. 6. Cytotoxicity Ė Another issue that comes up frequently when choosing among different gene. delivery methods is the cytotoxicity of the various methods and reagents. Some methods, such. as electroporation and gene guns, can be extremely harsh on cells and accuplacer often result in the death. of a majority of cells. Also, the DEAE-dextran can be quite harsh on term housekeeping some cell types, especially. when a DMSO or glycerol shock step is used as part of the procedure. In some applications, such as when a small number of transfectants are required from a recalcitrant cell type, this is.

not a problem. In the majority of applications however, a high percentage of cell death is simply. not acceptable. The plethora of s rice, available cationic lipids and polymer reagents have varying. degrees of toxicity with different cell types, and their concentrations during transfection can be. optimized as needed to minimize cell death. 7. Suspension vs. Adherent cells Ė Most commonly transfected cells grow Ė and are. transfected Ė while attached to some type of term about, support, whether that support is the teknik belajar mempengaruhi, tissue from. which they are derived, some inert membrane or scaffolding, or the surface of paper, a tissue culture.

plate. Less common are cells that grow while suspended in medium, such as those derived from. blood and other bodily fluids. Suspension cells are generally more difficult to writing in english sports transfect; therefore, it may be necessary to rely on harsher gene delivery methods, such as. electroporation, gene guns, microinjection, or viral vectors for acceptable transfection levels. Also, some cationic lipids can effectively transfect suspension cells, and should be tried when.

possible due to term paper housekeeping their simplicity and cost-effectiveness. 8. Expertise Ė The degree of difficulty in performing the various transfection techniques can. vary significantly, and test essay therefore the expertise and experience of the researcher is usually an. important factor to term housekeeping consider. For example, methods like microinjection viral transfection methods are invariably more complex than cationic lipids and polymer reagents. Also, of the chemical methods, calcium phosphate co-precipitation can be quite tricky due to its sensitivity to teaching slight changes in term, experimental conditions. 9. Time Ė Another important factor to consider is the time required for gene delivery.

Viral. methods, while highly efficient, can be quite time consuming, typically taking anywhere from two weeks to one month to complete. In contrast, chemical and physical/mechanical methods can usually be completed within a few days, but frequently provide lower transfection efficiencies. Thus, it is not uncommon to have to of internet essay weigh the benefits of faster results and less labor against the benefit of higher efficiency. 10 Cost Ė Finally, cost can be a major factor when deciding on the best gene delivery. method for term about, a given application. The most expensive methods tend to be the. physical/mechanical methods, such as electroporation and essay about teaching methods gene guns, due to the specialized.

equipment they require.Viral gene delivery is term paper perhaps the second most expensive method, since it requires the in english sports and games, vector reagents, extensive time and labor to complete lengthy vector preparation procedures, as well as proper disposal of viral waste materials. The third most expensive gene delivery methods utilize polymers and cationic lipids, which do not require any specialized equipment or lengthy viral vector preparation steps. Such reagents typically cost in housekeeping, the hundreds of dollars, depending on the number of transfections to test essay be completed. About? Finally, the least expensive methods utilize calcium phosphate, which is a cheap and writing abundant mineral, and naked DNA, which obviously requires no transfection reagent cost, but can be highly inefficient. GENE TRANSFER TECHNIQUES. Transformation is the naturally occurring process of gene transfer which involves absorption of the genetic material by a cell through cell membrane causing the fusion of the foreign DNA with the native DNA resulting in term about, the genetic expression of the received DNA. Writing In English Sports And Games? Transformation involves the uptake of ďnakedĒ DNA (DNA not incorporated into structures such as chromosomes) by competent bacterial cells. Cells are only competent (capable of taking up DNA) at a certain stage of their life cycle, apparently prior to the completion of cell wall synthesis . Transformation is usually a natural method of gene transfer but as a result of technological advancement originated the artificial or induced transformation.

Thus there are two types called as natural transformation and artificial or induced transformation. In natural transformation, the foreign DNA attaches itself to the host cell DNA receptor and with the help of the protein DNA translocase it enters the host cell. The presence of nucleases restricts the entry of two strands of the paper about, DNA, destroys a single strand thus allowing only about teaching methods, one strand to enter the host cell. Term Paper Housekeeping? Any DNA that is accuplacer essay not integrated into the chromosome will be degraded. This single stranded DNA mingles with the host genetic material successfully. The artificial or induced method of transformation is done under laboratory condition which is either a chemical mediated gene transfer or done by electroporation..

Genetic engineers are able to induce competency by putting cells in certain solutions, typically containing calcium salts. At the entry site, endonucleases cut the DNA into fragments of 7,000-10,000 nucleotides, and the double-stranded DNA separates into single strands. The single-stranded DNA may recombine with the hostís chromosome once inside the cell. This recombination replaces the gene in term about, the host with a variant Ė albeit homologous Ė gene. DNA from a closely related genus may be acquired but, in general, DNA is not exchanged between distantly related microbes.

Not all bacteria can become competent. Conjugation is a means of gene transfer in many species of bacteria. Cell-to-cell contact by a specialized appendage, known as the F-pilus (or sex pilus), allows a copy of an who can make essay F- plasmid (fertility plasmid) to transfer to a cell that does not contain the plasmid. Housekeeping? On rare occasions an F-plasmid may become integrated in debra s rice resume, the chromosome of its bacterial host, generating what is known as an Hfr (high frequency of recombination) cell. Such a cell can also direct the synthesis of a sex pilus. As the chromosome of the Hfr cell replicates it may begin to cross the pilus so that plasmid and chromosomal DNA transfers to the recipient cell. Such DNA may recombine with that of paper about, its new host, introducing new gene variants. Plasmids encoding genes for virulence factors antibiotic-resistance are passed throughout populations of bacteria, and between multiple species of bacteria by conjugation. In the writing sports and games, laboratory, conjugation can be used to transfer disrupted genes on a self-transmissible plasmid, to develop a mutant strain. A gene of housekeeping, interest from a recipient E. coli strain is cloned into a self-transmissible vector and maintained in a donor strain for genetic manipulation. The deleted gene construct is disadvantages of internet essay writing then transferred by conjugation from the donor strain back into the recipient strain.

Transduction is term housekeeping another method for transferring genes from one bacterium to another; the transfer is mediated by bacteriophages. Accuplacer? A bacteriophage infection starts when the virus injects its DNA into a bacterial cell. The bacteriophage DNA may then direct the synthesis of new viral components assembled in the bacterium. Bacteriophage DNA is replicated and term paper then packaged within the phage particles. Early in the infective cycle the who can make me an essay for university, phage encodes an enzyme that degrades the DNA of the host cell. Some of these fragments of bacterial DNA are packaged within the bacteriophage particles, taking the place of phage DNA. About? As the number of accuplacer test, phages increases it breaks open (lyse) the cell. When released from the infected cell, a phage that contains bacterial genes can continue to infect a new bacterial cell, transferring the bacterial genes.

Sometimes genes transferred in this manner become integrated into the genome of their new bacterial host by paper about homologous recombination. Such transduced bacteria are not lysed because they do not contain adequate phage DNA for viral synthesis and these phages can enter an alternate life cycle called lysogeny. In this cycle, all the virusís DNA becomes integrated into the genome of the host bacterium. Of Internet Essay Writing? The integrated phage, called a prophage, can confer new properties to the bacterium. Researchers use transduction as a means of gene transfer as it requires a media like virus for transferring genes from one cell to the other. Viruses are thus as a tool to introduce foreign DNA from the selected species to the target organism. There are two types of transduction called as generalized transduction in which any of the bacterial gene is transferred via the bacteriophage to the other bacteria and specialized transduction involves transfer of limited or selected set of genes. Viral vectors for gene transfer:

The use of viruses as vectors for transduction, i.e. the introduction of genes into animal cells by exploiting the natural ability of the virus particle, within which the transgene is packaged, to adsorb to term paper about the surface of the cell and gain entry is about methods being employed . Term About? Due to debra the efficiency with which viruses can deliver their nucleic acid into cells and term the high levels of replication. and gene expression it is essay possible to achieve, viruses have been used as vectors not only for paper about, gene expression in cultured cells but also for gene transfer to living animals. Four classes of viral vector have been developed for use in human gene therapy and have reached phase 1 clinical trials. These are the retrovirus, adenovirus, herpesvirus and adenoassociated virus (AAV) vectors . Before introducing the individual vector systems,we discuss some general properties of viral transduction vectors. Transgenes may be incorporated into. viral vectors either by addition to the whole genome or by replacing one or more viral genes. This is generally achieved either by ligation (many viruses have been modified to incorporate unique restriction sites) or homologous recombination.

For many applications, it is favourable to of internet essay use vectors from which all viral coding sequences have been deleted.These amplicons (also described as Ďgutless vectorsí) contain just the cis-acting elements required for packaging and genome replication. The advantage of such vectors is their high capacity for term, foreign DNA and test essay the fact that, since no viral gene products are made, the vector has no intrinsic cytotoxic effects. The choice of vector depends on about housekeeping the particular properties of the virus and essay writing in english the intended host, whether transient or stable expression is required and how much DNA needs to be packaged. For example, icosahedral viruses such as adenoviruses and retroviruses package their genomes into preformed capsids, whose volume defines the maximum amount of foreign DNA that can be accommodated. Conversely, rod-shaped viruses such as the baculoviruses form the capsid around the genome, so there are no such size constraints. There is paper housekeeping no ideal virus for gene transfer Ď each has its own advantages and disadvantages. In recent years, a number of hybrid viral vectors have been developed incorporating the beneficial features of essay, two or more viruses. 1) viral vectors that especially efficient at transducing the intended cell type but not other cell types. 2) retroviruses are particularly proficient at delivering genes to paper about housekeeping dividing cells, such as cancer cells or immortalized cell lines. 1) cannot deliver genes to terminally differentiated cells. 2) viral vectors may be highly immunogenic.

TRANSFECTION BY CHEMICAL METHODS. One of the methods of gene transfer where the genetic material is deliberately introduced into the animal cell in view of studying various functions of proteins and the gene is transfection. This mode of gene transfer involves creation of pores on essay the cell membrane enabling the cell to receive the foreign genetic material. Paper? When transformation is carried out in eukaryotic cells it is s rice resume termed as transfection. 1) Calcium phosphate mediated DNA transfer. The process involves a mixture of isolated DNA (10-100ug) with solution of. calcium chloride and potassium phosphate. Paper? . When the two are combined, a fine precipitate of the teknik pencapaian, positively charged calcium and the negatively charged phosphate will form, binding the DNA to be transfected on its surface .Cells are then incubated with precipitated DNA either in solution or in tissue culture dish. A fraction of cells will take up the calcium phosphate DNA precipitate by a process not entirely understood but thought to term about be endocytosis. Transfection efficiencies using calcium phosphate can be quite low, in the range of 1-2 % . It can.

be increased if very high purity DNA is essay teaching methods used and the precipitate allowed to form slowly. 1. Frequency is very low. 2. Term About? Integrated genes undergo substantial modification. 3. Many cells do not like having the solid precipitate adhering to them. and the surface of their culture vessel. 4. Integration with host cell chromosome is random. 5, Due to above limitations transfection applied to somatic gene therapy. This technique is used for introducing DNA into mammalian cells. This process has been a preferred method of identifying many oncogenes. 2)DNA transfer by DAE-Dextran method:

DNA can be transferred with the help of DAE Dextran also. DAE-Dextran may be used in the. transfection medium in mempengaruhi, which DNA is present. This is a polycationic, high molecular weight substance and is convenient for transient assays. The negatively charged DNA binds to the polycation and the complex is taken up by the cell via endocytosis.

If DEAE (diethylaminoethyl )-Dextran treatment is term paper housekeeping coupled with Dimethyl Sulphoxide (DMSO) shock, then upto 80% transformed cell can express the essay writing in english sports, transferred gene. The advantage of this method is that, it is cheap, simple and about can be used for transient cells. which cannot survive even short exposure of debra s rice, calcium phosphate. The major advantages of the technique are its relative simplicity and speed, limited expense, and remarkably reproducible interexperimental and intraexperimental transfection efficiency. Disadvantages include inhibition of cell growth and induction of heterogeneous morphological changes in cells. Furthermore, the housekeeping, concentration of serum in the culture medium must be transiently reduced during the transfection. It does not appear to be efficient for the production of stable transfectants. For cells that grow in suspension, electroporation or lipofection is usually preferred, although DEAE-dextran-mediated transfection can be used. Transfer of DNA by polycation-DMSO:

As calcium phosphate method of DNA transfer is reproducible and efficient but there is narrow range of optimum conditions. DNA transfer by. Another polycationic chemical,the detergent Polybrene is carried out to increase the adsorption of DNA to thesis belajar mempengaruhi pencapaian the cell surface followed by a brief treatment with 25-30% DMSO to increase membrane permeability and enhance uptake of DNA. In this method no carrier DNA is required and stable transformants are produced. This method works with mouse fibroblast and chick embryo. LIPOSOME MEDIATED GENE TRANSFER. Liposomes are spheres of lipids which can be used to transport molecules into term the cells. These are artificial vesicles that can act as delivery agents for exogenous materials including transgenes. They are considered as spheres of lipid bilayers surrounding the molecule to be transported and promote transport after fusing with the cell membrane.

Liposomes for use as gene transfer vehicles are prepared by test adding an appropriate mix of bilayer constituents to an aqueous solution of DNA molecules. In this aqueous environment, phospholipid hydrophilic heads associate with water while hydrophobic tails self-associate to term about housekeeping exclude water from within the lipid bilayer. This self-organizing process creates discrete spheres of continuous lipid bilayer membrane enveloping a small quantity of DNA solution. The liposomes are then ready to writing in english and games be added to target cells. Germline transgenesis is about possible with liposome mediated gene transfer.Lipofection is the most common and generally utilized gene transfer technique in the recent years and it utilizes cationic lipids. The combined DNA and debra resume cationic lipids act instantaneously to form structures called as lipoplexes that are more complex in structure than the simple liposomes. Cationic lipids have a positive charge and these form cationic liposomes which interact with the negatively charged cell membrane more readily than uncharged liposomes.

This leads to a fusion between cationic liposome and the cell surface resulting in quick entry by endocytosis and the delivery of the paper about housekeeping, DNA directly across the plasma membrane. Cationic liposomes can be produced from a number of cationic lipids that are commercially available and sold as an in vitro-transfecting agent, termed lipofectin. However this pathway would usually result in the fusion of lipoplexes with lysosomes and undergo degradation. This problem is overcome by utilizing the neutral helper lipids, which are generally included along with the cationic lipid. This allows entrapped DNA to escape the endosomes, reach the nucleus and get access to the cellís transcriptional machinery.The endosomal structure is destroyed by increasing the osmotic pressure created by the lipidsí buffering action within the endosomes and by thesis teknik mempengaruhi the fusion of the lipid with the endosomal membrane. The ability of a lipid to destroy endosomes is paper one of the main characteristics of a transfection reagent. 2. Test? Long term stability. 4. Protection of nucleic acid from degradation.

5. particularly proficient at delivering genes to dividing cells, such as cancer cells or immortalized cell lines. 6. cationic lipids effectively transfect suspension cells. 1) Cannot deliver genes to terminally differentiated cells. A rapid and simple technique for introducing genes into a wide variety of microbial, plant and term housekeeping animal cells, including E. coli, is electroporation. Electroporation uses electrical pulse to writing produce transient pores in the plasma membrane thereby allowing macromolecules into the cells. These pores are known as electropores which allow the about, molecules, ions and water to pass from one side of the membrane to another. The electropores reseal spontaneously and the cell can recover. The pores can be recovered only essay, if a suitable electric pulse is applied .The formation of electropores depends upon the cells that are used and the amplitude and duration of the electric pulse that is term paper about applied to them. When subjected to electric shock, cells take up exogenous DNA from the suspending solution.

Once inside the cell, the DNA is integrated and the foreign gene will express. A proportion of these cells become stably transformed and teknik belajar can be selected if a suitable marker gene is carried on the transforming DNA. The most critical parameters are the intensity and duration of the electric pulse, and these must be determined empirically for different cell types. Electric currents can lead to dramatic heating of the cells that can results in cell death. Heating effects are minimized by using relatively high amplitude, a short duration pulse or by using two very short duration pulses. However, once optimal electroporation parameters have been established, the method is simple to carry out and paper about housekeeping highly reproducible. Many different factors affect the who can me an essay for university, efficiency of about housekeeping, electroporation, including temperature, various electric-field parameters (voltage, resistance and capacitance), topological form of the writing sports, DNA, and various host-cell factors (genetic background, growth conditions) and post-pulse treatment. General applications of electroporation. 1. Introduction of exogeneous DNA into animal cell lines, plant protoplast, yeast protoplast and bacterial protoplast.

2. Electroporation can be used to increase efficiency of transformation or transfection of bacterial cells. 3. Wheat, rice, maize, tobacco have been stably transformed with frequency upto 1% by term housekeeping this method. 4. Genes encoding selectable marker may be used to introduce genes using electroporation. 5. To study the transient expression of molecular constructs. 6. Electroporation of early embryo may result in the production of transgenic animals. 7. Hepatocytes, epidermal cells, haematopoietic stem cells, fibroblast, mouse T and B lymphocytes can be transformed by this technique. 8. Naked DNA may be used for gene therapy by applying electroporation device on animal cells. Advantages of electroporation. 1. Method is fast. 3. Applied for essay, a number of cell types.

4. Simultaneously a large number of cell can be treated. 5. Term About? High percentage of stable transformants can be produced. 1)Limited effective range of. 1 cm between the electrodes. 2)Surgical procedure is required to place the electrodes deep into disadvantages essay the internal organs. 3)High voltage applied to tissues can result in irreversible tissue damage as a result of thermal heating. The technique has high input costs, because a specialized capacitor discharge machine is about housekeeping required that can accurately control pulse length and thesis teknik belajar mempengaruhi amplitude. 4) Larger numbers of cells may be required than for other methods because, in many cases, the most efficient electroporation occurs when there is up to 50% cell death. Sonication is the act of applying sound energy to agitate particles in a sample. Paper? Ultrasonic frequencies (20 kHz) are usually used, leading to the process also being known as ultrasonication . Sonoporation, or cellular sonication, is the use of these ultrasonic frequencies for modifying the permeability of the cell plasma membrane.

This technique is usually used in molecular biology and non-viral gene therapy in order to allow uptake of large molecules such as DNA into the cell, in a cell disruption process called transfection or transformation. Sonoporation employs the acoustic cavitation of microbubbles to enhance delivery of such large DNA molecules. Although the early results could achieve only test, 10- to 30-fold increases in transfection efficiency in vitro, technical refinements have lead to enhancements of up to several thousand fold in vitro ,sufficient to encourage ultrasonication mediated gene transfer in vivo. 1) It is non-invasive and term housekeeping well tolerated, 2)extraordinary safety record over a wide range of frequency and intensity. 3)high levels of public acceptability and understanding. there are highly sophisticated, flexible, cost-effective and readily available diagnostic and therapeutic systems that can achieve site-specific transfer of essay methods, ultrasound energy almost anywhere in the body, except perhaps the about, lung. 4) The bioactivity of this technique is thesis mempengaruhi pencapaian similar to, and in some cases found superior to, electroporation. 5) in vivo is term housekeeping relatively nontoxic.

1)Extended exposure to low-frequency (MHz) ultrasound has been demonstrated to result in complete cellular death (rupturing) as well as microvascular hemorrhage and disruption of tissue structure, thus cellular viability must also be accounted for when employing this technique. 2) Whether ultrasound affects later steps in the transfection process, particularly plasmid entry into the nucleus, remains unclear. Sonoporation is under active study for the introduction of foreign genes in tissue culture cells, especially mammalian cells. Sonoporation is also being studied for essay writing in english sports and games, use in about, targetedGene therapy in vivo, in a medical treatment scenario whereby a patient is given modified DNA, and essay about methods an ultrasonic transducer might target this modified DNA into specific regions of the patientís body. In microinjection DNA can be introduced into cells or protoplast with the help of very fine needles or. glass micropipettes having the diameter of 0.5 to 10 micrometer. The DNA may be directly delivered into the nucleus or in the cytoplasm. Some of the DNA injected may be taken up by the nucleus.

The desired gene in the form of plasmid or alone is injected directly into the plant protoplast. Injection into the nucleus is paper more efficient than that into who can me an the cytoplasm. Paper About Housekeeping? Micro-injection is carried out with automatic equipments (robotics) using a micro-needle. Computerized control of holding pipette, needle, microscope stage and video technology has improved the efficiency of this technique. Advantages of microinjection. 1. Frequency of stable integration of DNA is accuplacer essay far better as compare to other methods. 2. Method is effective in transforming primary cells as well as cells in paper about, established cultures. 3. The DNA injected in this process is subjected to less extensive modifications. 4. Mere precise integration of essay writing, recombinant gene in term about housekeeping, limited copy number can be obtained. 2. Skilled personal required.

3.Has not been successful for many plant cells . 4. Teaching? Embryonic cells preferred for manipulation. 5. Paper About Housekeeping? Knowledge of mating timing, oocyte recovery is essential. 6. Method is useful for protoplasts and essay about teaching methods not for the walled cells. 7. Process causes random integration. 8. Rearrangement or deletion of host DNA adjacent to site of integration are common. 9. The technique is laborious, technically difficult, and limited to the number of cells actually injected. Applications of microinjection. 1. Process is applicable for plant cell as well as animal cell but more common for term paper, animal cells. 2. Technique is ideally useful for producing transgenic animal quickly. 3. Procedure is thesis mempengaruhi pencapaian important for gene transfer to embryonic cells.

4. Applied to term paper housekeeping inject DNA into plant nuclei. 5. Method has been successfully used with cells and protoplast of. tobacco, alfalfa etc. Fig: microinjection in thesis mempengaruhi pencapaian, mouse egg. Biolistics or particle bombardment is a physical method that uses accelerated microprojectiles to. deliver DNA or other molecules into intact tissues and term cells. Biolistics transformation is relatively new and novel method amongst the physical methods for artificial transfer of exogenous DNA. The nucleic acid is delivered through membrane penetration at a high velocity, usually connected to microprojectiles like microscopic particles of tungsten or gold coated into cells using an electrostatic pulse, air pressure, or gunpowder percussion. As the particles pass through the essay teaching, cell, the DNA becomes free to integrate into the plant-cell genome.

The gene gun is term housekeeping a device that literally fires DNA into target cells . Essay About Teaching Methods? The DNA to be transformed. into the term paper, cells is coated onto microscopic beads made of either gold or tungsten. Beads are carefully. coated with DNA. The coated beads are then attached to the end of the plastic bullet and loaded into. the firing chamber of the gene gun. An explosive force fires the bullet down the barrel of the gun. towards the target cells that lie just beyond the end of the teknik, barrel.

When the term about, bullet reaches the end of. the barrel it is caught and stopped, but the essay about teaching methods, DNA coated beads continue on toward the target cells. Some of the beads pass through the cell wall into the cytoplasm of the target cells. Paper About Housekeeping? Here the bead and. the DNA dissociate and the cells become transformed. Once inside the disadvantages writing, target cells, the DNA is. solubilised and term paper about housekeeping may be expressed. 1. Biolistics technique has been used successfully to transform soyabean, cotton, spruce, sugarcane, papaya, corn, sunflower, rice, maize, wheat, 2. Genomes of subcellular organelles have been accessible to in english sports genetic. manipulation by biolistic method. 3. Mitochondria of term, plants and chloroplast of about, chlamydomonas have been.

4. Method can be applied to filamentous fungi and paper about yeast (mitochondria). 5. The particle gun has also been used with pollen, early stage. embryoids, meristems somatic embryos, plant cells, root section, seeds and pollen. 6. This approach has been shown to debra s rice resume be effective for transferring transgenes into mammalian cells in vivo . 1. Requirement of protoplast can be avoided. Unlike electroporation and microinjection, this technique does not require protoplasts or even single-cell isolations and is applicable to even intact plant tissue.

2. Paper Housekeeping? Walled intact cells can be penetrated. 3. Manipulation of genome of subcellular organelles can be achieved. 4. This is who can make essay also a highly mechanized or robotic mediated technique in which the. speed of the micro-projectile particles is controlled by foolproof mechanisms. 5) It is also gaining in use as a method for term about housekeeping, transferring DNA construct into whole animals.

6) This technique is fast, simple and safe, and it can transfer genes to a wide variety of tissues. 7)there appears to be no limits to the size or number of genes that can be delivered. 1. Integration is random. 2. Who Can Me An Essay For University? Requirement of equipments. 3.It can be a challenge to obtain a sufficient number of cells modified by term housekeeping this method to writing in english sports and games see a biologically significant effect. A new technique is presented to incorporate exogeneous gene materials (DNA) into cells with a microbeam irradiation from an about uv pulsed laser. A frequency-multiplied Laser, 355 m wavelength, 5 ns pulse duration, punches a self-healing hole of submicrometer aperture in thesis teknik, cell membrane under selected irradiation conditions.

At the site of the beam impact, due probably to local temperature changes, the cell membrane modifies its permeability. As a consequence of the hit, circular areas, whose radius may be apparently regulated by changing the irradiation time and/or the radiation intensity (energy), appear on the wall, last for a short time and fade spontaneously . It takes a fraction of housekeeping, a second for the aperture to close, long enough to allow the foreign DNA, contained in the medium, to who can make me an slip into term about housekeeping the cell. It is well established that a strong pressure wave, known as a laser-induced stress wave (LISW), accompanies laser-induced plasma. Essay Teaching Methods? We have extended their method to deliver macromolecules, such as genes. Plasmid DNA (circular DNA residing in bacterial cytoplasm) is used as a vector of the gene of interest. We inject the plasmid into target tissue, on which a laser target is placed, and subject the target to irradiation with a high-intensity, nanosecond laser pulse to induce plasma and hence an LISW.

By placing optically transparent material on term about housekeeping the target, the plasma is confined, resulting in an increase in of internet writing, the LISWís impulse. Interaction of term paper about, tissue cells with the LISW allows plasmid to enter the test essay, cytoplasm. 1) it only paper about housekeeping, takes advantage of the presence of phenol-red, a normal cell medium component, with no need of addition of for university, extraneous substances; (2) it is a very mild treatment virtually suitable for any cell type and. (3) it allows transfection of selected cells even in the presence of paper about, cells of s rice, different type (providing that they are morphologically distinguishable). 4) . it is rapidly replacing virus mediated techniques as they have serious side effects caused by immune response and limited targeting characteristics rendering them inappropriate for clinical application. 5) The method offers a clear advantage over existing methods: increases the paper housekeeping, success rate of teknik belajar, DNA transfection as well as the efficiency of term about, cell modification by orders of magnitude. 6) . It enables minimally invasive tissue interaction. Thus there are a number of debra, ways by which the genes can be introduced into the cells. With the advent of molecular tools and technologies it is now comparatively easy to introduce gene into term about housekeeping cells without losing its integrity and biological activity.

Moreover the recent development in molecular biology has made the and games, transfer of term paper, gene with great accuracy to the target cell. The transfer of gene through different gene transfer technologies has cured a number of diseases. Research is on who can progress to housekeeping cure those diseases which cannot be cured by using drugs. Moreover the treatment of diseases by gene transfer provides better result for a prolong period of time. It is the need of hour to discover new and accuplacer test essay cheap method of term, gene transfer technologies so to make the treatment of the diseases a little easier and affordable.

Improvements in gene transfer are required in terms of transfection approaches to allow improved transgene uptake efficiencies. 1)Society of Applied Sciences. Gene Transfer Technologies and their Applications: K.H.KHAN. Medical Biotechnology Division, School of Biosciences and Technology, VIT University, Vellore-632014, Tamil Nadu, India. 2)Principles of gene manipulation: R.W.Old ,S.B. Primrose and R.M.Twymann Ď sixth edition,chapter 10 and 11. 3) BIOTECHNOLOGY 2020-From the essay about teaching, Transparent Cell to. the Custom-Designed ProcessĖProf.

Gerhard Kreysa Dr.-Ing. Housekeeping? E.h. Essay Teaching Methods? Dr.h.c. Term? Dr. R??diger Marquardt. 4) INTRODUCTION TOBIOTECHNOLOGY AND GENETIC ENGINEERING.

5)From genes to of internet writing genomes- concepts and term about housekeeping applications of of internet writing, DNA technologyĖJeremy W Dale, Malcom von Schantz. Search our thousands of essays: If this essay isn't quite what you're looking for, why not order your own custom Science essay, dissertation or piece of coursework that answers your exact question? There are UK writers just like me on hand, waiting to help you. Each of us is qualified to a high level in our area of expertise, and term paper housekeeping we can write you a fully researched, fully referenced complete original answer to your essay question. Just complete our simple order form and you could have your customised Science work in your email box, in as little as 3 hours. This Science essay was submitted to and games us by a student in order to help you with your studies. This page has approximately words. If you use part of this page in term, your own work, you need to provide a citation, as follows: Essay UK, Essay: Gene transfer techniques Ė pros and cons . Available from: [04-10-17]. If you are the thesis teknik mempengaruhi, original author of this content and no longer wish to have it published on our website then please click on the link below to request removal:

13-09-17 - The Human Genome project (HGP) 22-08-17 - Adeno associated virus (AAV) vectors 01-08-17 - Test methods on the alkali carbonate reaction in concrete - fluxonic diode - Identifying an unknown organism from a cell phone 05-07-17 - Risk factors for alcohol abuse in men who have sex with men 01-06-17 - Monsanto vs. the world 19-05-17 - Patterns formed by increasing the scale factor Ė maths 12-05-17 - Swine influenza infections 24-04-17 - Medicinal plants and wound healing. We offer professional custom essay writing, dissertation writing and about coursework writing service. Our work is high quality, plagiarism-free and delivered on time. Essay UK is a trading name of Student Academic Services Limited , a company registered in who can me an, England and Wales under Company Number 08866484 .

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Custom Mangrove Forest Project essay paper writing service. According to Orr (2008), Al-Dhakira Mangrove forest covers an term paper housekeeping area of 4 kilometer of the coast from south to north, and about 2 kilometers at the farthest points, from east to west. The specific site of the project is located just opposite Al-Dhakira town. Teknik Belajar! He notes that the ecosystem has dense mangrove Avicennia marina, mudflats, especially, those which are broad in nature, and finally the salt-marsh vegetation. The bay is surrounded by a spit right from the term about open sea up to the east, which has offered it a kind of natural protection.

Its long drift runs from the north to the south. Additionally, it has a good spawning and who can make, nursery area which has been utilized economically for rearing some valuable fish and even the shrimp stocks. The area, therefore, supports artificial fishing, while also providing grazing land, especially to camels which feed on the mangroves. Orr (2008) further indicates that Al-Dhakira is part of the Qatar Peninsula which is paper, mainly flat and looks like a stony desert. Qatar is among the areas in of internet the region with the very harsh climates. It is normally extremely dry and receives a very low annual rainfall which is usually less than 130 mm. The area has long summers which are mostly hot and of high humidity. Its winters, on the other hand, are normally cool during the day, while it is paper, freezing during the night. Writing In English! Being a desert, its temperature ranges from 23oC to 35oC during cold and warm seasons respectively. Spalding, Kainuma Collins (2010) argue that with the extreme climatic conditions in this area, it has not been easy for vegetation to thrive.

They are not even where there is soil widespread, but they are normally extremely shallow and mostly made of sandy loams which are often overlaid with varying sizes of rock debris. This type of soil cannot allow the paper about housekeeping people in the area to practice agriculture. Equally there is limited possibility of the growth of natural vegetation and wildlife with just some scattered vegetation to the North. Its wetlands are majorly marine and coastal. It is, therefore, the who can me an for university mangrove trees that live on salty water and can survive in term paper about this harsh climate. This has also been the accuplacer essay case, especially in term paper about housekeeping the areas surrounding the coast where Al-Dhakira, the east coast, is covered with a widespread of mangroves. However, Orr (2008) notes that the about teaching methods area has only term about housekeeping one species of mangrove, which has been described as the essay writing sports black mangrove or the Avicennia marina . There have also been attempts by the government to plant mangroves, though its effort has been limited to small areas in the local regions. Today, Al-Dhakira area is surrounded with the term paper forest of mangrove trees which makes it an ideal home for certain types of fish, organisms and sea birds. Spalding, Kainuma Collins (2010) noted that even though the area has not been comprehensively studied, a few studies conducted in the area have shown that the area is fit for wintering and the passage of water birds. It is, therefore, inhabited by such birds as Platalea leucordia and accuplacer essay, the Sterna anaethetus . Paper About Housekeeping! The site can, therefore, generate tourism inside. However, up to this time the forest has not gained enough publicity at both the individual citizen and institutional levels.

This has been identified as the major cause of lack of attention to this ecosystem which has remained as one of the pillars of the essay about teaching surrounding maritime life in Qatar. According to Spalding, Kainuma Collins (2010), the term problem has been the lack of information and proper study of educate people and institutions on this profound element of me an for university life in the area. Orr (2008) confirms that extensive researches on the existing literature have proved futile because there is no current information regarding the current state of vegetation in the area. Term Paper! Such details like an estimate of the vegetation cover and the records of the changes that have been taking place over time cannot be easy traced. He argues that the publications have also failed in providing the estimates of the area under vegetation around the small town. This is a clear indication that there has not been a comprehensive survey of the area under the who can Al-Dhakira.

This can lead to true loss of about housekeeping marine life and who can make me an essay, eventually to more damage to the environment. In fact, mangrove forests have become an important part of the people’s life in the small town of Al-Dhakira. Orr (2008) observes that the area is term about housekeeping, currently a source of both coastal and essay writing in english, marine resources, including fish. The management of coastal resources is, therefore, a very key aspect of this ecosystem, as it is connected directly to people’s welfare. The site has been protected with booms, especially whenever oil spills occur. However, Orr (2008) notes that it has not been formally recognized by paper housekeeping, the government. This means that it has not been put under protection as one of the essay writing key areas for conservation of wildlife in the country. The area has therefore faced a number of problems which seemingly will worsen in the future. First among such problems is the issue of camel grazing that has remained a major challenge to the sustainability of the mangrove ecosystem. Second, the land is term about, bound to face the challenges related to the ongoing growth of a new town currently coming up to the north coast. Orr (2008) warns that without appropriate interventions, there will be a problem of land reclamation and recreational disturbances in essay writing in english sports the near future.

Another problem that has always affected the site is that one of the oil spill. Unfortunately, there has been very little if any to mitigate the effects of these factors in paper about the future. The impact of oil, for example, would have been so adverse if it could not have been for the topographical nature of the teaching methods site which has provided a good protection around it. Term Paper About Housekeeping! The effect would worsen in the future with 90% of the population of Qatar presently living in its towns and cities. The Starting Point. Having looked at the current situation, it is clear that there is a great need for writing in english and games an urgent action towards the protection of Al-Dhakira Mangrove forest. There are various issues that will be considered as a key to the starting and the realization of the goals of this project.

First, this project will have to identify the real problems facing mangrove forests in Al-Dhakira, the actual partners, the customers, and the beneficiaries. The project will then establish the type and volume of the work it would take for the project to be implemented. The project will also aim at learning from what other similar projects, carried out in other countries, are facing. According to Spalding, Kainuma Collins (2010), even though there have been various attempts to establish mangrove plantations, much has not been achieved. Term About! He observes that the only areas which have realized some level of success are the protected inlets which exist along the east coast. The areas lying to the west coast have particularly lagged behind because of their high level of water salinity. Even though humans in Qatar have not ventured into accuplacer major usage of these forests, they sometimes carry out certain activities like recreational boating and fishing.

Just like the other countries from the Middle East, one of the term paper housekeeping problems facing Al-Dhakira has been the browsing of thesis belajar pencapaian domestic animals which has not posed much threat because the pressure has not been so much allowing the term about forest to re-grow. However, Orr (2008) notes that oil spills is the greatest problem, as it causes a great pollution. Equally, there has been a problem of coastal development whose effect has been on resume the increase, as it has resulted into term an ever increasing reclamation to accommodate the excess population migrating into these areas. Additionally, he observes that the resume growth of about housekeeping these coastal towns has also led to increase damping of solid and liquid waste in mangrove forests. Unfortunately, the country has not demarcated any coastal area for protection. However, there have been numerous proposals seeking for the protection of Al-Dhakira through declaring it a biosphere reserve.

In addition, Qatar Petroleum has equally aiming at protecting the mangroves in Qatar. In comparison to what is happening in Saudi Arabia which has sparse mangroves on the coasts of both the Persian Gulf and Red sea. The country also faces the about challenges of high salinity and the browsing of camel and commercial. About! Mangroves have also been used here to of internet essay writing attract tourists, especially those who would want to participate in bird watching. The country also faces the term paper about housekeeping challenge of reclamation of s rice resume land for urbanization and industrialization. Equally oil pollution has also been a threat, especially along the Arabian Gulf; an example occurred during the 1991 first Gulf War. However, unlike Qatar, the country has recognized the contribution of these areas and various measures have, therefore, been put in place towards rescuing the mangrove from the effects of term paper housekeeping these threats. Such measures include expanding of the shrimp farming and maintenance and protection of the mangrove fringes, especially those near the ponds. The country has also declared four areas protected.

Additionally, Orr (2008) noted that there had been the promotion of plantation of mangrove in small scale. Such efforts have been put forward by such bodies as the National Commission for Wildlife Conservation and who can make me an, Development which together with other parties like the various oil companies have been supporting the Project of the Wetland Protection and Mangrove Rehabilitation. The country has also experimented with the plantation of a number of alien species along the Northern Gulf coast. Having determined the current situation, the team will prepare a questionnaire and housekeeping, conduct an initial feasibility study. The entire period of the study will take one month. Thesis Teknik Pencapaian! This one month will be divided to enable all the stages of term about research to be accomplished. These include: seeking for permission from the relevant authorities, development of the questionnaires which fit every target group, training of those who would take part in the administration of the questionnaires, carrying out a pretest of the tools, conducting the actual research, analyzing the findings, and finally presentation and dissemination of the findings. The study will consider as its sample population people from among those already taking part in the Mangrove Project in who can me an essay for university Qatar and other people who will be drawn from the larger Qatar.

The team will then analyze the information collected from the field and come up with a report which will clearly outline all the details and plan on how the project will be implemented. At the beginning of the project, the term about team shall hold an official meeting which will bring together all the stakeholders in the project. This project is timely and very relevant as it will not only complement the existing projects on the ground working to manage the resources along the Qatar coast, but it will also help in bringing together the stakeholders to ensure that their intervention is of great impact. The project has been necessitated by the worsening situation of the mangrove ecosystem at Al-Dhakira, even amidst the government claiming that it had put the area under protection. The participants in this project shall include: all the about teaching Qataris, Al-Dhakira population, Qatar schools, the research team which is running the prevention project, fishermen, tourists and most of the institutions and the companies in Qatar. Identifying the Problems. The attempt to conserve the mangrove forest in Al-Dhakira has seen a number of challenges with the major problem being the issue of the salinity of the water in the surrounding area. This has resulted from the construction of term paper fish-pens which has caused disturbances to the normal flow ebbing the tidal water and navigation.

This in turn affects the filtration process which is always made possible by the presence of the mangrove forest. Once affected, the forest loses its effectiveness in filtering the wastes originating from the communities staying off the shore. Mempengaruhi Pencapaian! This results into siltation and, thus, lowers the quality of water. Many fish are, thus, killed making the economic state of the community even worse. The issue is further complicated by the fact that the water in Al-Dhakira has kept on term paper housekeeping rising, creating an ecological imbalance that is accuplacer test, not conducive for the growth of the mangrove. Paper About Housekeeping! Together with the essay writing problem of the term paper about scarcity of marine life which has majorly been caused by the effects of the large construction projects, located just along the coastal waters, Qatar is slowly losing its mangrove forest in thesis belajar mempengaruhi pencapaian the Al-Dhakira area. Projects Goals and Objectives.

The goal of this project is to protect mangrove trees. Term Paper! This will be achieved majorly by debra s rice, creating awareness through public education to lobby all the term about housekeeping people of Qatar to participate in the protection and attraction program. The project also aims at building an information center with a view of ensuring sustainability through presenting the available information materials to all the people of Qatar, thereby creating a knowledgeable population. Additionally, the project aims at strengthening the accountability of the institutions involved through training them in the management of the mangrove ecosystems. Lastly, the project also aims at increasing the disadvantages of internet writing level of participation of the community towards the management of the housekeeping mangrove ecosystem. The project will employ a number of strategies which have been well outlined.

These strategies will include conducting a detailed study. This shall be done by the consultants. The report will then be analyzed for me an a better understanding of the present situation of the ecosystem. Term Paper! Another strategy will be to organize series of teaching methods presentations to the responsible parties, such as the relevant government officials, project personnel, university students and even school pupils. The project team will also seek to gain approval for about housekeeping constructing scenario for protecting the mangrove trees. Essay Writing Sports And Games! To ensure sustainability of the impacts of the project, the team will seek to ensure the building of an information center which shall be responsible for running the forest. The centre will include a small team to protect the forest and provide information while at the same time carrying out term about housekeeping continuous studies to update the same information. Gathering Information and Requirements. The information shall majorly be obtained from the administration of questionnaires and the deliberations that shall be reached at by the disadvantages of internet team of stakeholders. The team shall use the results from the analysis of the feedbacks from the questionnaires and that of the deliberation by the project team to come up with useful information that the team can use to guide its campaign on paper the need to protect and manage mangrove forests. However, such strategies, as chosen by the team, will then have to be scrutinized to accuplacer test essay allow the identification and adoption of the most useful ones.

In the process of reviewing the strategies, the term team will visit schools and universities to solicit for belajar mempengaruhi pencapaian their support as well as to provide the relevant information to them. With this kind of interaction, the team will be able to term paper about test effectively its strategies. The team will also conduct workshops during which it will be possible to define various issues related to mangroves with the stakeholders being encouraged to disadvantages come up with their own strategies towards addressing such challenges. The media will also be contacted to provide an opportunity which can be utilized by the stakeholders concerned with educating the public to pass various environmental related information issues, especially those directly linked to the management of mangrove ecosystem. The community will be provided with such messages during the various meetings which shall be open for all the community members. The community members are also important during the implementation stage of the project.

The community may include such issues as the need to avoid the construction of fish pond along the mangrove ecosystem. There is need for the community to understand that such developments can adversely affect their own natural resource base and cause economic crisis, as it will affect the supply of all the resources obtained from that ecosystem. Finally, the team will also approach the decision makers to get the initial approval. This stage will also include the intentions/ideas for expansion or additional services to be provided at the location. The project institutionalization will be strengthened by persuading various institutions to commit financially as part of about their support to the project. The success factors in the study will include: protecting the ecosystem, providing the right environment for who can make me an for university the marine life, introducing a new attraction place for about the tourists inside and outside Qatar, protecting the surrounding land against the possibility of the rise of sea level, and preventing the area from the temperature rise. Identifying Expectations and Risks.

There is no true present risk, apart from the cost, that will be directly incurred in carrying out the activities that aim at achieving the objectives of the project. The scope of the work will include the initial consultant’s presentation, initial agreement, setting up the team members, estimating the sports and games workload expected and the budget required. Term Housekeeping! The scope will also clearly state if there are other people involved, the location and the number of field trips to essay be conducted. The stakeholders include all Qataris, Al-Dhakira populations, Qatar Schools and the research team which is running the prevention project. The research team will also be responsible for carrying out a rapid resource assessment to know the extent to which the damage of the ecosystem has occurred. Other stakeholders will include the fishermen, tourists and most of the term institutions and companies in Qatar. The project will also work together with municipal authorities along the coastal strip. Whether or not the approval is about teaching, granted will depend on term paper housekeeping the benefits that are expected from the project. There will be the management responsible for writing in english sports achieving the objectives. They will approve the project and assign the team their duties to ensure that the about goals are realized. Finally, at this stage, the consultancy team will kick start the project by collecting the information, developing the next move and providing the necessary recommendations.

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Essay on Why Cannabis Should be Legalized. Naturally, I had to write about marijuana! Itís written in a variation of APA format and I thought I might as well submit it so some of our readers could read itÖmaybe steal it for an assignment youíre putting offÖfeel free to critique as much as you want; after all, I am a student and term, that is essay in english and games, how I learn. And I know I could have expanded a LOT more but it was just a simple school essay. Also, some things, like the Heath/Tulane Study, may have some variations, but I found tons and tons of different information and I tried to compile the info I felt was most credible. Enjoy! Marijuana is a very prominent and controversial issue in society today. Although many slanderous claims have been made about cannabis in recent history, the truths are slowly starting to term paper, resurface. Unfortunately, these truths are under heavy criticism due to the stereotypical view of what people view as the typical ďpot smoker.Ē This skewed perception of about a lazy and unmotivated America is the result of over seventy years of propaganda and misinformation spread by private interests who needed cannabis illegal for their own personal gains. As a former marijuana consumer and current administrator for a marijuana activism website, I strongly believe that marijuana should be legalized for all uses. Using marijuana for medicinal purposes is term, not a recent discovery.

In fact, written references to use medical marijuana date back nearly 5,000 years. Western medicine embraced marijuanaís medical properties in the mid-1800s, and by the beginning of the 20th century, physicians had published more than 100 papers in the Western medical literature recommending its use for essay a variety of disorders ( Accessed on June 5, 2010). These disorders include pain relief, nausea, glaucoma, and movement disorders. It is also used as a powerful appetite stimulant which helps patients that had HIV/AIDS or cancer patients going through chemotherapy. Today, thousands of patients are able to term paper about housekeeping, use marijuana as an effective method of essay treatment for their ailments. Paper? This is possible because fourteen state governments across the country have acknowledged cannabisí effective healing properties and legalized its consumption for medicinal purposes with the consent of writing a licensed physician.

However, these patients are at the mercy of the federal government and about housekeeping, its authority. One of the biggest positive impacts that the legalization of disadvantages marijuana would produce is the reintegration of hemp into our society, and paper, more importantly in current times, our economy. Hemp is who can make for university, a plant that is term housekeeping, grown for industrial use only; in fact, hemp contains less that 1% THC and causes no ďhighĒ when smoked. From an test, industrial standpoint, hemp provides many advantages over a great deal of current resources that America utilizes. The most common use for hemp is in the production of textile based products. Hemp fibers are considerably strong so that makes it ideal for term paper housekeeping the production of many products like paper, fabric, and rope. Hemp produces a higher yield per acre than cotton and has a growing cycle of only 100 days instead of 160. Hemp seeds provide more protein than soy, and according to the U.S. Department of Agriculture, you can produce four times as much paper from an acre of marijuana than an acre of trees; plus, marijuana can be harvested at who can make essay for university a quicker rate than trees ( Accessed on June 5, 2010). But perhaps the term most important attribute of hemp is its ability to accuplacer test, produce a substantial amount of cellulose. Cellulose is a compound that, in more recent years, has been converted into a biofuel called cellulostic ethanol.

This biofuel can power everything that gasoline currently does and is being produced most commonly in paper about housekeeping, the form of ďenergy crops;Ē mostly corn and of internet essay, cotton. Hemp holds an advantage though, because it can be produced more frequently than cotton, and term paper about, it yields four times the debra s rice amount of cellulose you can get from a corn stalk ( Accessed on June 5, 2010). In todayís economic situation, hemp would be a huge revenue creator. Currently, the paper about housekeeping United States has to import all of its hemp from countries like Canada and China, who have no laws banning the cash crop. Not only would domestic hemp production stop the pumping of American dollars into foreign farmersí pockets, but it would create a mass amount of jobs for otherwise unemployed Americans. Essay And Games? In these times of uncertainty, we need jobs and we need more domestic industry, and hemp can accomplish both of those.

Recent times have also shown us the damage that we are doing to our environment and the realization that we canít depend on fossil fuels anymore. With hemp being a major contributor to the alternative fuels movement, it makes one wonder why itís still illegal to paper about, grow it. Because of the recent focus on marijuana legalization, many people are looking back to s rice, why cannabis was criminalized in term paper about housekeeping, the first place. Contrary to popular belief, the of internet essay reason marijuana was made illegal had nothing to term paper about, do with health issues. Essay Writing In English And Games? The real reason was simple, good old fashioned, American business. As the methods for housekeeping processing hemp into make essay, paper and plastics were becoming more readily available and affordable, business leaders including William Randolph Hearst and about housekeeping, DuPont stood to lose fortunes.

They were faced with either spending massive amounts of money adapting their prominent, timber-based, industries to produce hemp, or be run out of business by essay writing the conveniences and low prices that came with these hemp producing innovations. Paper Housekeeping? Instead of doing either of these, they both did everything they could to s rice, make their competition illegal. This wasnít as hard as one might think because lucky for Hearst, he also owned a chain of well known newspaper chains. DuPontís chief financial backer Andrew Mellon was responsible for appointing Harry Anslinger in 1931 as the term paper housekeeping head of the debra s rice Federal Bureau of housekeeping Narcotics and Dangerous Drugs. Hearstís papers routinely fabricated stories about ďmarijuana-crazed black menĒ raping white woman, and accuplacer, with these types of horrifying acts reported all over the media, Anslinger testified before congress that, ďMarijuana is the most violence causing drug in term about, the history of disadvantages of internet essay mankind.Ē ( Accessed on June 7, 2010).

Later in history, during the term housekeeping Red Scare, Anslinger again testified to congress against marijuana, but this time he stated that marijuana caused people to become peaceful and passive, and the communists would use it to weaken Americaís will to fight. Another major factor to the criminalization of marijuana (and the term ďmarijuanaĒ itself), was racism. In the 1930ís the Great Depression hit hard, and at the same time an influx of Mexican immigrants were coming into America. Unfortunately, just like modern times, in the 1930ís these immigrants were not greeted with open arms. Just like they are today, Mexican-Americans were accused of stealing work from ďhonest, hard-working Americans.Ē Many of these immigrants smoked a Mexican weed called ďmarihuana.Ē Marihuana was not cannabis. It wasnít until William Randolph Hearst used the name for the smokable form of cannabis in his ludicrous newspaper articles that people started to associate Mexicans with the drug, too. People supported making the about methods drug illegal because if the Latin-American people were caught consuming it, they would be deported back to their country of origin.

As a kid growing up, youíre constantly told of the term paper dangers of accuplacer essay drugs, and since it is illegal, marijuana is a drug and therefore it is dangerous. Again, the ďdangersĒ of term paper housekeeping marijuana can be traced back to the 1930ís and the negative propaganda that surrounded it to debra s rice resume, appease private interests and reinforce racial stereotypes; never on actual dangers to your health. Term? In 1974, Dr. Robert Heath of Tulane University conducted a test in which lab monkeys were given thirty marijuana joints daily to thesis teknik pencapaian, test for harmful side effects. After 90 days, many of the monkeys started to die and the university reported that the monkeys had died from paper about a mass dying of accuplacer test essay brain cells and they concluded that the consumption of marijuana destroyed the brain. The official test observations and results, however, were withheld from the public for six years until finally, in 1980, they were released to NORML (The National Organization for the Reform of Marijuana Laws) due to a lawsuit NORML had brought to the university.

Upon further investigation of the study, it was revealed that Dr. Heath actually pumped the equivalent of term paper about housekeeping sixty-three joints via gasmasks into the monkeys everyday in just a five minute time frame; much more than the first reported thirty joints a day. The monkeys were reported to have died because of accuplacer test essay brain damage, but in reality they died of suffocation; when the body doesnít get enough oxygen, one of the first things to term, start dying off are brain cells ( Accessed June 7, 2010). To this day, there has been no test like the Heath/Tulane study. In 2006, alcohol contributed to 85,000 deaths in the United States. In 2000 alone, tobacco contributed to a staggering 435,000 deaths!

In the entire history of mankind, marijuana has never been recorded as a cause of death. No one in recorded history has died from the consumption of cannabis ( Accessed on June 7, 2010). However, marijuana has been linked to short term memory damage and to a slightly reduced lung capacity when regularly inhaled. Although no where near as severely as alcohol, marijuana does also impair judgment and who can essay for university, motor skills. As is the same with most other things, those who regularly ingest cannabis usually build up a tolerance to these side effects which makes them even less hindering, but some individuals never fully develop one. Marijuana is also not as addictive as most media outlets would lead you to believe.

The most addictive drug, even above heroin, is nicotine. Housekeeping? After nicotine and heroine comes cocaine, and s rice resume, then alcohol. Below that is caffeine, and at the very bottom of the paper housekeeping chart is marijuana; less addictive than your can of Dr. Accuplacer? Pepper ( Accessed on June 7, 2010). Marijuana laws, as they are now, cost tax payers billions of dollars a year.

The taxpayers have to pay for every individual that is sent to prison for possession of marijuana and nothing more. Most people are sent to prison for possession because they are on parole and it is a violation to about, have any illegal drugs on disadvantages essay you or in your system. Because of the three strike rule, they are only three joints away from spending life in prison, which is extremely expensive for the taxpayers; all because of marijuana remaining an illegal drug. Taxpayers also have to term paper about housekeeping, pay for every bit of essay litigation that leads up to the ďoffenderísĒ incarceration. It is estimated by a Harvard study, that over $7 billion is spent annually on marijuana prohibition ( Accessed on June 7, 2010). If cannabis were legalized, and then regulated and taxed like other commercial goods, it would bring in huge dollar amounts to the federal government. Itís estimated that the tax revenue generated from marijuana would be upwards of $2.4 billion annually if it were taxed like everyday goods, and as high as $6.2 billion annually if it were taxed like alcohol and tobacco ( Accessed on June 7, 2010). There are many arguments against the legalization of marijuana; many of which I have already addressed.

There are some arguments that have more to do with social problems than with personal values or health issues. One prominent argument against term paper housekeeping marijuana legalization is that if cannabis is made legal, then it will get into the hands of our children. This has been proven untrue; there is even a counter-argument that states it is in fact easier for make me an essay children to get marijuana than it is for them to obtain alcohol. 35% of students said marijuana was easily obtainable while only housekeeping, 5% said alcohol was easy to obtain ( Accessed on June 7, 2010). Many people, including politicians, have agreed with this data taking into account that drug dealers donít ask for I.D. Probably the biggest argument against marijuana is that itís a ďgateway drug.Ē Many people claim that consuming marijuana greatly increases your chance of doing harder drugs like cocaine and heroin. This is a textbook example of a ďpost hoc ergo propter hoc fallacy.Ē This informal fallacy occurs when one assumes the temporal order of events supports their casually being related. Just because most people that do hard drugs started with marijuana does not mean that marijuana will cause people to do harder drugs. If someone wants to experiment with cocaine they are going to disadvantages, do it regardless if they have tried marijuana before. Term Paper About Housekeeping? People need to stop putting the blame of heavy drug users onto this harmless plant.

No one wants to take responsibility for thesis mempengaruhi their actions and own up to their mistakes; everyone looks for the scapegoat. By this logic, you shouldnít eat meat because it leads to cannibalism. Also, itís never reported that cigarettes are a ďgateway drug.Ē How many people do you think tried smoking a cigarette or drinking alcohol before smoking marijuana? By the ďgateway drugĒ theory it should be cigarettes and alcohol that are leading people to meth and about housekeeping, heroin, not marijuana. Itís hard to see why cannabis is still illegal in mempengaruhi, the country. Paper About? To this day, I have not heard an of internet essay writing, intelligent argument against the legalization and regulation of cannabis that cannot be quickly dismissed or refuted.

Most politicians, I would assume, are worried that supporting this controversial subject would yield negative feedback from their supporters. The public has gotten it into their heads that legalizing marijuana is the same as condoning it, which it is not. One doesnít have to term about housekeeping, condone the smoking of marijuana to vote for itís legalization. The fact is, many people would outlaw fast food, cigarettes, alcohol, and tanning beds because of the harm they cause. Disadvantages Essay? But this is America, and America is about freedom and about choice. Give the people in our nation the choice to enjoy marijuana (which is not nearly as dangerous as those four things I just mentioned) without the fear of prosecution or criticism. In these economic times the government should be looking for ways to create jobs, produce ongoing revenue, and increase morality of its citizens. Why wonít they overturn this unjust law?

Canít they just expand their minds? (1999, September 3). CASA Study Finds Marijuana Easier for Teens to Get Than Beer. Retrieved June 7, 2010, from term about housekeeping : old/106/casastudy.shtml. (2008). Essay About Teaching? Annual Cause of Death in the United States. Retrieved June 7, 2010, from Drug War Facts:

(2010, January 30). Introduction to Medical Use. Term Paper About? Retrieved June 5, 2010, from debra s rice resume NORML: Herer, J. (2009). The Emperor Wears No Clothes.

Retrieved June 7, 2010, from term housekeeping Washington Free Press: Miron, J. (2005, June). The Budgetary Implications of belajar pencapaian Marijuana Prohibition. Retrieved June 7, 2010, from : Murphy, K. Housekeeping? (2009, June 3).

How Marijuana Became Illegal. Make? Retrieved June 7, 2010, from Washington Free Press: (n.d.). Which Drugs are the Most Addictive?. Retrieved June 7, 2010, from Schaffer Library of Drug Policy: Urena, A. (2002, November 25). Legitimate Reasons to Legalize Cannabis. Retrieved June 5, 2010, from housekeeping Cannabis News:

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