Wistar Preparation Report

The Helsinki declaration was founded in 1964 and since here they have been updating principles of good practice in this kinds of experiments. They say, “Welfare of animals used for research must be respected”. Since here the ethical views for non-humans subjects started. Later on the APA gave instructions to determine whether an experiments on animals is acceptable or not. For example animal research should not harm the animal or distress it on any way. If the researchers don’t follow this code, they wont be allowed to keep on with the experiment.

Li, and Huang discuss the controversial topic of gene therapy by first providing a factual summary of gene therapy. Their introduction goes into the specifics of the types of vectors (carriers that deliver a gene to target cells) used to transfer genes in order to aide damaged proteins. Afterwards, they transition into the detailed of process of gene therapy. They start this off with naked DNA and state it is the simplest approach to nonviral transfers and the least toxic method. However, despite being the least toxic and a nonviral transfer it does not have as great of an effect as Cationic lipids. Cationic lipids have been used in various experiments but are known to be safe only when administered in small doses. Furthermore they go into

Gene therapy is a relatively new technology that has shown some promising results. Such as the drug called Strimvelis that is used to help children with severe combined immunodeficiency, a disease that leaves infants with little to no protection against viruses, bacteria, fungi, and other potentially harmful things. It appears to be a cure carried out through a genetic pair. Strimvelis was tested on 18 children, starting 15 years ago, all of whom are still alive today. Gene therapy is used to inject genes into a person’s cells and tissues in order to treat a disease, as well as hereditary diseases where an abnormal allele is replaced with a fully functional allele. The carrier, called a vector, is used to transport the normal gene to the target cells. The most common type of vector are viruses that have been genetically altered to carry human DNA. Scientists have been trying to control the way the viruses have evolved encapsulating and delivering their genes to normal cells in a pathogenic way by manipulating the viral genome to remove disease-causing genes and inserting healthy ones. Theoretically, it is possible to transform either the cells of the body (somatic) or the cells of the sperm, ova, and their precursors (germline). All of the gene therapy experiments so far on humans have been directed to somatic cells, because germline experiments have been remaining controversial. Somatic gene therapy is split into two broad categories: ex vivo (cells are modified outside of the body then returned back again) or in vivo (genes are changed in cells while still located inside of the

Gene therapy is a rapidly developing technology first used in 1990.1 It involves the use of modified DNA inserted into an organism with a missing or mutated sequence. There are two methods of delivering this correction into the patient’s cells. In the viral method, a virus containing the vector with the corrected form of the gene, infects the cell naturally. Adenoviruses, adeno-associated viruses, lentiviruses, and retroviruses can all be used. They each utilize a different method of delivery and form of DNA. Adeno-associated viruses and lentiviruses are the easiest forms to construct.2 The corrected DNA faces many obstacles to gene expression and ultimately protein production. The viruses must target and enter the correct cell, avoid

Gene therapy is deemed to be the ‘use of genes as medicine’. “Gene therapy is a form of therapy that involves inserting one or more corrective genes that have been designed in the laboratory, into the genetic material of a patient's cells to cure a genetic disease by replacing the missing or malfunctioning genes” (Mandal). There are two types of gene therapy, which are in vitro gene therapy, which takes place outside the body and in vivo therapy, which takes place inside the body. Both types are very time-consuming processes containing many detailed steps that can easily go wrong. First is in vitro or ex vivo gene therapy, which again is outside the body. In this technique, cells are surgically taken out from the tissue area that is impacted. The next step is that the corrective DNA or genes that will be used to treat the disease are spliced or injected into the cells, and then the cells are left to be cultured. The final step is that the new tissues are put back into the body in the area, which was affected. The use of bone marrow cells as recipients of the gene is very common due to the fact that they create the blood that will eventually be circulated throughout the body (Oracle ThinkQuest).

Total proteins were prepared from cells of embryonic cortex at different ages (E10–18) and adult brain (2 months) with 500 µl of cell lysis buffer. Cell lysis buffer contains 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 1 mM Na2EDTA, 1 mM EGTA, 1% Triton, 2.5 mM sodium pyrophosphate, 1 mM β-glycerophosphate, 1 mM Na3VO4, 1 μg/ml leupeptin, 2 µg/ml aprotinin, and 1 mM PMSF. After homogenization on ice for 10 min, the supernatant of each sample was collected and stored in aliquots at -80 °C. For each sample, the protein concentration was determined by Bradford assay (Thermo Fisher Scientific). Cell lysate (20–50 µg) was resolved on 12% SDS-polyacrylamide gel electrophoresis and transferred onto PVDF or nitrocellulose membranes. The protein blots were blocked with 5% milk in TBS (10 mM Tris–HCl, pH 8.0, 150 mM NaCl) overnight at 4 °C and incubated with primary antibodies at a dilution of 1 to 1000–2000 in 5% milk prepared in TBS. The secondary antibody was donkey anti-mouse IgG-700 or anti-rabbit IgG-800 (LI-COR). Quantitative western blot was detected by the Odyseey CLx Infrared Imaging System according to the company’s instructions

Administration of poly herbal capsules did not produce any change in serum electrolytes such as sodium, potassium, chloride and bicarbonate indicating a normal electrolyte balance is maintained during poly herbal capsules administration (Table 4).

As years go by and technology advances, many medical techniques that failed in the past are starting to seem more possible. Researchers are discovering that a technique called gene therapy is able to help prevent and reverse the effect of many conditions. Originally in the early 1900's, gene therapy failed in

In the last two decades, gene therapy has improved from mental concepts to clinical trials and laboratory experiment. Gene therapy is a technique that is currently being tested to see if genes can be used to treat or prevent disease. This method can successfully be able to replace a mutated gene with a healthy copy, knock out a mutated gene that is not functioning and introduce a new type of gene in the body to help fight a disease a human being may have. Gene therapy is a work in progress system that basically inserts a “normal” genome to replace the gene that is “abnormal” and causes disease.

Gene therapy is an experimental treatment, which is used to replace mutated, improperly functioning genes with regular DNA. There are several gene therapy methods, most of which involve using a vector to transport the corrected gene into targeted cells. Retro viruses, adeno viruses, adeno ascosciated viruses and liposomes are commonly used as

Human Gene Therapy was first written about in 1963 with the first clinical study using gene transfer being conducted in 1990. Human Gene Therapy is characterized by the insertion of a functioning gene into cells to fix a cellular dysfunction or to create new cell function (Culver 1994). There are many diseases that come from dysfunctional or defective genes. These diseases include cystic fibrosis, combined immunodeficiency syndromes, hemophilia, and many types of cancer. (Cotrim & Baum 2008). A gene can be distributed to a cell using a carrier known as a vector. The most common vectors used in gene therapy are viruses. The viruses

There is a variety of ways that animal experimentation can be conducted and there are many purposes for it. The following major fields of use may be acclaimed: basic biomedical research, applied biomedical research, regulatory testing of drugs, compounds and products, routine testing of biological substances and products, and educational purposes (Kolar). Basic research is used to investigate veterinary or medical implications as well as biological phenomena. Almost anything can be done to the animals during these

Animal experimentation has existed since ancient times and contributed to human life and survival. The most radical progress in the health science has all been made possible by animal research. Thus, animal experimentation is recognized and accepted as an important tool in saving lives.

The coverslips were washed with 1X PBS. Permeabilized for 5 minutes with 500 µl PBS + 0.2% Triton X-100, then washed again with 1X PBS. The cells were blocked for 1 hour with 2% BSA in PBS. Then the cells were incubated for 1 hour with primary LAMP1 antibody raised in mice at a concentration 1:50 (cat. ab25630 Abcam), diluted with 2% BSA in PBS. The coverslips were washed 3 times with 1X PBS and incubated for 45 minutes with secondary Alexa Flour 647 goat anti mouse antibody (cat. A21236 Invitrogen), at a concentration of 1:500, diluted in PBS + 2% BSA. The coverslips were then washed 3 times with 1X PBS. The coverslips were mounted using DAKO mounting media (cat. S3023), and imaging of the cells was performed using an LSM 710 (ZEISS) confocal fluorescence microscope.

Gene therapy treats and prevents a disease by introducing a vector of genetic material into certain cells to alter the function or ability of a gene. The promise of gene therapy as a cure for diabetes has been considered ever since this new technology emerged into the clinical and research sphere. Although such methods have yet to undergo human clinical tests, gene therapy holds much potential to bring a radical new way of treating autoimmune diseases such as diabetes. By targeting certain genes that control the insulin and ?-cell production in the pancreas, gene therapy will someday fulfill its potential to cure the disease that is the number one cause of heart disease in the