SF-9 Lepidopteran Cells Essays - Benzofurans, Biochemistry Methods

SF-9 Lepidopteran Cells Essays - Benzofurans, Biochemistry Methods SF-9 Lepidopteran Cells PHM499 Research Project Supervisors: Dr. P. S. Pennefather, Dr. S. M. Ross Calcium transport study of SF-9 lepidopteran cells and bull frog sympathetic ganglion cells Kenny Yu Faculty of Pharmacy, University of Toronto, 19 Russell Street, Toronto, Ontario M5S 2S2 ABSTRACT The intracellular calcium level and the calcium efflux of the bull-frog sympathetic ganglion cells (BSG) and the SF-9 lepidopteran ovarian cells were investigated using a calcium-sensitive fluorescence probe fura-2. It was found that the intracellular calcium levels were 58.2 and 44.7 nM for the BSG cells and SF-9 cells respectively. The calcium effluxes following zero calcium solution were 2.02 and 1.33 fmolecm-2s-1 for the BSG cells and SF-9 cells. The calcium effluxes following sodium orthovanadate (Na2VO4) in zero calcium solution were 6.00 and 0.80 fmolecm-2s-1 for the BSG cells and the SF-9 cells. The SF-9 cells also lost the ability to extrude intracellular calcium after 2-3 applications of Na2VO4 while the BSG cells showed no apparent lost of calcium extruding abilities for up to 4 applications of Na2VO4. INTRODUCTION Spodoptera frugiperda clone 9 (SF-9) cells are a cultured insect cell line derived from the butterfly ovarian tissue. SF-9 cells are used by molecular biologists for the studies of gene expression and protein processing (Luckow and Summers, 1988). However, there is not much known about these cells' basic biophysiology. Since calcium is involved in many cells' activities such as acting as a secondary messenger, it is important for cells to control their intracellular calcium level. This study was aimed toward looking at the some of the basic properties of the SF-9 cells such as resting calcium concentration and rate of calcium extrusion after being calcium level being raised by an ionophore 4-bromo-A23187. The effect of sodium orthovanadate (an active transport inhibitor) on calcium extrusion was also looked at. Microspectrofluorescence techniques and the calcium-sensitive probe fura-2 were used to measure the intracellular calcium concentration of these cells. In addition, the BSG ce lls were used to compare with the SF-9 cells for the parameters that were studied. It was found that the SF-9 cells appeared to have a calcium concentration similar to the BSG cells. Moreover, the calcium extrusion rates of both cell types with no Na2VO4 added seemed to the same. However, due to insufficient data, the effects of Na2VO4 could not be statistically analyzed. From the data available, it suggested that the BSG cells' rate of calcium extrusion was enhanced by the Na2VO4 and was greater than the SF-9 cells. It was more important to note that the calcium extruding capabilities of the SF-9 cell seemed to impaired after two to three applications of Na2VO4 but it had apparent effects on the BSG cells even up to 4 applications. After obtaining these basic parameters, many questions raised such as how does the SF-9 cells extrude their calcium and why the Na2VO4 affected the calcium efflux for the SF-9 cells but not the BSG cells? The SF-9 cells may have a calcium pump or exchanger to extrude their calcium and they may be very sensitive to the ATP (adenosine 3'-triphosphate) supply. This was apparently different from the BSG cells' since their calcium extrusion were not affected by the Na2VO4.. It may be useful to find the mechanism(s) of the actions of Na2VO4 on the SF-9 cells because it may find possible applications in agriculture such as pest control. MATERIALS AND METHODS Chemicals and solutions 4-bromo-A23187 and Fura-2/AM were purchased from Molecular Probes (Eugene, OR). Na2VO4 was purchased from Alomone Lab (Jerusalem, Israel). Dimethyl sulfoxide (DMSO) was obtained from J. T. Baker Inc. (Phillipsburg, NJ). All other reagents were obtained from Sigma (St. Louis, MO). The normal Ringer's solution (NRS) contained (mM): 125 NaCl, 5.0 KCl, 2.0 CaCl2, 1.0 MgSO4, 10.0 glucose, 10.0 N-[2-hydroxyethyl] piperazine-N'-[2-ethanesulfonic acid] (HEPES). The calcium free Ringer solution (0CaNRS) is the same as the NRS except CaCl2 was substituted with 2.0 mM ethylene glycol-bis(b-aminoehtyl) ether N,N,N',N'-tetraacetic acid (EGTA). Fura-2/AM solution was prepared as follows: a stock solution of 1mM fura-2/AM in DMSO was diluted 1:500 in NRS containing 2% bovine albumin. It was then sonicated for 10 minutes. It was then kept frozen until the day of the experiment. 20 SYMBOL 109 f "Symbol"M 4-bromo-A23187 solution was prepared by diluting a stock of 5mM 4-bromo-A23187 in DMSO 1:250 with NRS. Na2VO4 solution

Foods that Increase Brain Activity and Prevent Decline by EssaySupply.com

Foods that Increase Brain Activity and Prevent Decline Foods that Increase Brain Activity â€Å"Smart foods,† they’re called – foods that enhance brain activity and that keep the brain healthy as the body ages. We now know, from research, that certain elements in some foods have a positive impact on the brain. While much of this testing has occurred with rats, the implications for human brains are certainly there. The elements that are now known to improve brain health and to help prevent brain health decline are Omega-3’s, glucose, blood supply, and anti-oxidants. In fact, the research shows that oxidants are more detrimental to the brain than to any other organ in the body. If you would like detailed information about these studies, there are load of essays online that summarize them. Nevertheless, foods rich in these elements should be added to everyone’s diet right now, no matter what age you are. Blueberries: Studies show that this superfood improves learning capacity and motor skills in animal studies. As well, they protect the brain from oxidants, and more recent studies indicate that there may be properties that retard dementia and Alzheimer’s. Recommended daily amount is 1 cup. Wild Salmon: Omega-3 fatty acids are known to promote good brain function and to reduce or delay effects of aging. These are also found in sardines and herring, and the recommended amount is at least one service 2-3 times a week. Nuts and Seeds: These foods are really high in Vitamin E, a nutrient that is also known to retard the aging of the brain as we age. 1 ounce per day of most any type of nut plus sunflower and sesame seeds is recommended. Vitamin E is also found in peanut butter if that is preferred. Avocados: One of the best benefits of avocados is that they contribute to good blood flow. The brain, like any other organ, relies on a strong blood supply. This food also lowers blood pressure, a condition which, over time, is a factor in the decline of cognitive abilities. Recommended amount is  ¼ -  ½ of one avocado once a week. This food is very high in fat content and calories, so this is the reason for the lower recommended amount. Whole Grains: Everyone should eat at least 3 servings a day of whole grain foods – oatmeal, whole grain cereals, and whole wheat or rye breads. First, these foods lower cholesterol, a leading cause of plaque buildup in the arteries. This buildup decreases blood supply to the brain, and can cause a stroke or heart attack. Both of these events disrupt the flow of blood to the brain. Whole grains also stimulate blood flow, so there is a double-plus to eating the recommended amount. Beans: The brain relies on glucose, and it does not store this nutrient. So, it is important that a steady supply of glucose get to the brain so that it is maintained in a healthy condition. While beans do not of themselves have glucose in them, they do regulate the flow of glucose throughout the body and ensure that the brain gets an adequate supply. Everyone should eat  ½ cup of beans every day. Pomegranates and other bright/dark colored vegetables and fruits. Pomegranates are mentioned specifically because, of all fruits and vegetables, they have the highest amount of anti-oxidants. Other fruits and vegetables include broccoli, tomatoes, oranges, dark berries and grapes, dark lettuce, carrots, asparagus, and squash. Red wines are also full of antioxidants. 5 servings a day please. Tea: If it is freshly brewed tea (no instant!), there are moderate amounts of caffeine which does enhance focus and memory. Tea, especially green tea, also contains anti-oxidants. 2-3 cups a day is ideal. Dark Chocolate: Yes! This is a huge source of anti-oxidants, and it also have moderate levels of caffeine. A recommended daily amount is  ½ - 1 ounce each day. Hopefully, there are some foods on this list that you really like. If so, and there are others you just can’t abide, at least increase the amounts of those foods that you do like. Scientists have located a protein in the brain that is definitely linked to Alzheimer’s, so keeping that protein functioning properly is important. â€Å"Brain foods† will help!