INTRODUCTION: Apoptosis is a distinct form of a programmed cell death ( PCD) and defect in apoptosis are now thought to contribute to the development and progression of cancer. Regarding the keen interest in programmed cell death in the last years, more and more assays and commercial kits are emerging to prove apoptosis. However, their detectability and reliability have been often discussed. Here we introduce rapid and simple method for evaluating apoptosis in cancer researches and genotoxicities. METHODS: NIH-3T3 cell line were used in this study, after treatment with apoptotic agent, DNA was extracted by simple protocol. Then DNA ladder and flow cytometry assays were done for detection of apoptosis in NIH-3T3 cell line. RESULT: Primary and late apoptosis in the treated cells was determined by flow cytometry analysis. Accordingly DNA ladder assay using 1.5 % gel electrophoresis showed fragmentation in the DNA of treated cells. CONCLUSSION: This research provides a fast, simple and cost-effective method in observing apoptosis in mammalian cells and could be use in cancer research and genotoxicity.
Introduction
Apoptosis is a distinct form of a programmed cell death ( PCD) or cell suicide , first described by Kerr et. al. in the 1970s(1, 2). It is a normal physiological phenomenon that plays an important role in embryonic development, maintenance of tissue homeostasis and pathology(3) . Apoptosis triggered by exogenous and endogenous stimuli as radiation, oxidative stress and genotoxic chemicals. It is defined by characteristic changes in the nuclear morphology including chromatin condensation and fragmentation , overall cell shrinkage and formation of apoptotic cell bodies. Among the many markers of apoptosis, DNA fragmentation is...
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...spite benefits of the technique , that protocol cannot be used for other purposes such as PCR , because of presence of high amount of DMSO (7).DNA ladder assay is an easily available method and seems to be very useful for quick screening of apoptotic changes in cell populations. This method allows working with cell lysates and does not require any special laboratory equipment(10).The present study results showed that this DNA ladder assay used here, is a useful method for evaluating DNA damage and fragmentation caused by apoptotic agents , drugs , food additives and etc.
Conclusion
Although flow cytometry is a quantitative technique for determining apoptosis, we believe that this simple and rapid DNA ladder Assay will be very useful for numerous labs that routinely study cell death or carry out routine experimental/clinical screening of drugs and chemotherapeutics.
Montaner, B. and Perez-Tomas, R. 2001. Prodigiosin-induced apoptosis in human colon cancer cells. Life sciences, 68 (17), p. 2025-2036.
Euthanasia is one of the most complicated issues in the medical field due to the debate of whether or not it is morally right. Today, the lives of many patients can be saved with the latest discoveries in medicine and technology. But we are still unable to find cures to all illnesses, and patients have to go through extremely painful treatments only to live a little bit longer. These patients struggle with physical and psychological pain. Dr. Martin Luther King Jr. discusses the topic of just and unjust laws in his “Letter from Birmingham Jail” which brings into question whether it is just to kill a patient who is suffering or unjust to take that person’s life even if that person is suffering. In my opinion people should have the right, with certain restrictions, to end their lives in the way they see fit if they are suffering from endless pain.
We concluded that it is possible to examine small amounts of DNA by first replicating the sample using Polymerase Chain Reactions then using gel electrophoresis to. determine the genotype of the DNA. The main goal for our experiment was to learn how to examine DNA when there is only a small sample present. We examined the samples of DNA obtained from student hair using the replication method of Polymerase Chain Reaction and the inspection method of gel. electrophoresis.
Antonie Phillips Van Leeuwenhoek, the father of microbiology is one of the most influential people of science; But how and why?
3. Prospects for Antisense Nucleic Acid Therapy of Cancer and AIDS. Eric Wickstrom, Ed. Wiley-Liss, Inc., NY, 1991. pp 25-33, 35-51, 125-141.
Sophisticated methods of testing are now being applied to human cells in petri dishes. Human volunteers are also being used and micro-dose with samples so small that they do not cause adverse reactions. The argument exists that these alternative testing methods are not only more cost effective but also more relevant because they are conducted using human cells and specimens; a method that isn’t hindered by species differences. In addition, computer generated models are being used to produce virtual reconstructions in order to test toxicity.
The Phase I trial will be discussed here as it pertains to the topic at hand. The typical treatment for cervical cancer if surgery is not a viable option – like if the cancer has spread, then called locally advanced cervical cancer – is chemotherapy and radiation treatment at the same time. This phase I clinical trial is simply looking to add ipilimumab to this regimen, but once the chemo/radiation has been completed (LACC article). Chemo and radiation destroy tumor cells, which causes tumor-associated antigens to be released. Once released, these antigens are exogenous (outside the cell) and will be presented to helper T cells to initiate an immune response.
Telomere are special DNA structure that consist of repetitive nucleotide sequences, which serves as a “cap” to protect the ends of the chromosomes. These repetitive sequences can range from thousands of base pairs in Vertebrates to about a few hundreds of base pairs in yeast cells (Oeseburg, et al. 2009). Located at the ends of the chromosomes, the telomeres serves as a biological life line for cells. Once the telomeres reach a certain length, the cell will cease to divide. Oeseburg, et al (2009) suggested that the telomere has a crucial length, once reached, it could result in chromosome end-to-end fusion and chromosome dysfunction; which may eventually lead to cell apoptosis, c...
As an inducer of HIF-1 production, it’s been used to study the apoptotic effects in HepG2 cells.
The subject of death and dying can cause many controversies for health care providers. Not only can it cause legal issues for them, but it also brings about many ethical issues as well. Nearly every health care professional has experienced a situation dealing with death or dying. This tends to be a tough topic for many people, so health care professionals should take caution when handling these matters. Healthcare professionals not only deal with patient issues but also those of the family. Some of the controversies of death and dying many include; stages of death and dying, quality of life issues, use of medications and advanced directives.
A cellular signaling pathway has four major parts that determine its function. The initial signal, ranging in size and composition, is the first component in the signal. This will then determine what type of signal can occur and which receptor can accept the signal. Once the signal reaches the receptor, either intracellular or extracellular, the receptor responds by altering in order size/shape in order to accept the signaling ions. With completion of the cellular signal, the message is spread throughout the cell, causing short or long-term effects.
In Buddhism, eternalism and annihilationism are both considered heretical beliefs. The concept of each was brought up through the 10 Unanswered Questions, those which the Buddha would not respond to because they violated the core beliefs. The two questions that deal specifically with eternalism and annihilationism are “is the body one thing and the soul another?” and “is the soul the same as the body?” Through the presupposition of the existence of the soul present in both questions, to answer would be to acknowledge the existence of a soul and therefore, the belief in a self. These questions also refused the Doctrine of No-Self, by supporting the idea of eternalism, and the concept of rebirth, by supporting the idea of annihilationism. Buddhism seeks the middle-way between these two extremes that may appear to be contradictory. The Doctrine of No-Self and Dependent Origination are utilized in the explanation of the middle-way and, as a result, of Nagarjuna’s concept of emptiness. When the concept of radical interdependence, exhibited in the Doctrine of Emptiness, is understood then it reveals the understanding of
Euthanasia is the painless killing of a patient suffering from an incurable and painful disease or in an irreversible coma. Euthanasia can either take the form of passive or active assisted-suicide. Euthanasia is a hotly debated topic international that receives a lot of media attention when a story breaks about a personal story of someone suffering from an incurable and painful disease asks to be euthanized.
This philosophical study will define the good aspects of death that is revealed through the monotony of life over extended periods of time. In my view, death is a good thing because the concept of “self” presumes that I will remain unchanged if I was to live as an immortal. over time, the concept of I would become stagnant, and i would soon wish to kill myself out of boredom. Bernard Williams is an important premise for this belief, since the assumption of immortality usually rests on the assumption of maintaining the “self” over long periods of time. I do not believe that this motive is substantial in the desire to love forever, which would make death a good thing in terms
Cell death is not a random process, but occurs is a very organized programmed sequence of molecular events. This programmed cell death is called apoptosis. Apoptosis is a very clean death as compared to that of necrosis, where the cells swell and burst spilling their contents over their neighbors and eliciting an inflammatory response. Apoptosis is mediated by proteolytic enzymes called caspases, which cleave certain intracellular proteins to help kill the cell. There are two major classes of caspases initiator and executioner. Initiators as their name implies, iniate the apoptotic process and executioners catalyze cell wide protein cleavage events that will kill the cell. Apoptosis is a cascade event meaning there are multiple activation