The Impact of Temperature on Permeability of Cell Membrane - Assignment Example

?The impact of temperature on permeability of cell membrane and Contents 3 This study is a lab experiment of the impact of temperature on permeability of cell membrane. A cell membrane is a very important component of a cell because it is the one that controls movement of substances in and out of a cell, hence protecting it from its environment. The experiment will be performed on a beetroot cell. To further explore on the subject, recent experiments that have been performed with a similar purpose to the current study will be reviewed critically, and their findings linked to the findings of the current study. Most of these experiments focus on the way different levels of temperature can cause damage to a cell membrane, and hence causing it to rapture or to fail to carry out its functions properly. Prediction of the outcome of the study is that permeability of a cell membrane is increased with the increase in temperature, but up to a certain level of temperature increase, the cell membrane is damaged. At very high temperatures, the biochemical structures of a cell can be dented, but the most susceptible part is the cell membrane and that is why it will be the focus of this experiment. 3 The study will apply a controlled experiment to conduct the tests. This method is preferred especially because it can easily establish the cause and effect through manipulation of certain variables while keeping the others variables constant. Furthermore, lab experiment allows for accurate control of variables. Beetroots have been chosen as part of the study’s ethical consideration of using animal in experiments only as a last resort. After the experiment, the data will be recorded in an excel worksheet and later analysed using different methods, some generated by software and others done manually. These include descriptive statistical analysis, explanatory analysis, and use of graphs, scatter plots and charts. 3 Introduction/Background 4 Hypothesis: 6 Aims of the study: 6 Materials and methods 6 Materials 6 16.Methods 7 19.Procedures 8 23.Ethical considerations 10 25.Data analysis 10 73.Timeframe 11 97.References 12 Abstract This study is a lab experiment of the impact of temperature on permeability of cell membrane. A cell membrane is a very important component of a cell because it is the one that controls movement of substances in and out of a cell, hence protecting it from its environment. The experiment will be performed on a beetroot cell. To further explore on the subject, recent experiments that have been performed with a similar purpose to the current study will be reviewed critically, and their findings linked to the findings of the current study. Most of these experiments focus on the way different levels of temperature can cause damage to a cell membrane, and hence causing it to rapture or to fail to carry out its functions properly. Prediction of the outcome of the study is that permeability of a cell membrane is increased with the increase in temperature, but up to a certain level of temperature increase, the cell membrane is damaged. At very high temperatures, the biochemical structures of a cell can be dented, but the most susceptible part is the cell membrane and that is why it will be the focus of this experiment. The study will apply a controlled experiment to conduct the tests. This method is preferred especially because it can easily establish the cause and effect through manipulation of certain variables while keeping the others variables constant. Furthermore, lab experiment allows for accurate control of variables. Beetroots have been chosen as part of the study’s ethical consideration of using animal in experiments only as a last resort. After the experiment, the data will be recorded in an excel worksheet and later analysed using different methods, some generated by software and others done manually. These include descriptive statistical analysis, explanatory analysis, and use of graphs, scatter plots and charts. The impact of temperature on permeability of cell membrane Introduction/Background Permeability of cell membrane is the ability of a cell’s plasma membrane to allow substances to pass through it. The purpose of these substances, passing through the membrane, is to allow removal of waste products and entry of chemicals that are needed for the cell to carry out its functions in the body. On the other hand, cell membrane is the tissue that detaches the inside of cells from their surroundings (Alberts et al., 2002). The cell membrane is made up of hydrophobic and hydrophilic regions. The hydrophilic region, which is on the exterior of the cell, is attracted by water while the hydrophobic region is in the interior of the cell and does not come into contact with water. While the outer surface of the membrane is responsible for selecting the substances that can enter the cell, the interior surface is responsible for controlling the proteins that are used in the functions of the cell and its development. The chief purpose of a cell membrane is to control movement of substances in and out of a cell, hence protecting the cell from its environment (Budin and Devarai, 2011). This is made possible by the lipid and protein components that facilitate its functions. Besides protection of the cell, the cell membrane can also facilitate cell recognition, allow organelle motility and allow transport of substances in and out of the cell. A beetroot cell has anthocynin red pigments, which are contained in a large vacuole. This pigment cannot leak out of the vacuole because it is protected by the cell membrane. However, the cell membrane can rupture and allow the vacuole to free the anthocyanin, following damage of the cell membrane. If the membrane is damaged, it can destroy the structure of permeability and the proteins (Imgrund, 2009). Transportation of proteins in and out of the cell is carried out by the proteins, and when they are denatured, they fail to function properly. Furthermore, when proteins are subjected to severe temperature, the veracity of the cell membrane is diminished. This mean those substances that are not supposed to leave the cell can leave hence killing and denaturing the protein (Gamper, 2009). A number of studies have been conducted, investigating the extent of cell damage due to excessive temperature (Henriques, 1947; Leyko and Bartosz, 1986). Some of the experiences that have been discussed include denaturation of membrane, increase in membrane permeability, blebbing of membrane, cell lysis and collapse of cytoskeleton proteins. At very high temperatures, the biochemical structures of a cell can be dented, but the most susceptible tissue is the cell membrane and that is why this will be the focus of this experiment (Aveyard, 2010). The impact of thermal stress has been quantified through a number of studies that have investigated the changes of membrane in cells and liposomes. Kanehisa and Tsong (1978) conducted such an experiment and found that when loposomes with thermal alterations experience optimum permeability when in state of liquid-crystalline alteration of lipid bilayer. A similar finding was established by Cruzeiro Hansson et al. (1989) in their theoretical lipid membrane approach. Nonetheless, a definitive explanation of cell membrane, in the case of cells, transforms in the course of hyperthermia – quantification of this has been extremely difficult. By use of a kinetic of membrane bledd creation in Hela cells, Moussa et al. (1977) carried out an experiment with the aim of finding out the state of thermal damage –they found that very high temperatures damages the cell membrane and prevent it from performing its functions. Borrelli et al. (1986) in a more recent study have demonstrated that the distribution and size of a membrane blebbing induced with hyperthermia related with cell fatality at 14.50C. Padanilam et al. (1994) and Cravalho et al. (1992) investigated the damage of plasma membrane that was induced with thermal, using fluorescent dye leakage. Nonetheless, these studies did not have a comprehensive analysis of the real-time of cell membrane changes at physiological temperatures. Blinks conducted a study whose focus was investigating the resistance of membrane; hence establishing the extent of cell membrane permeation activation energies. Hypothesis: When temperatures are increased, beetroot cell membrane will move at a faster rate, hence increasing the permeability of the membrane. As such, it is expected that the relationship between temperature and permeability will be positive. In addition, the two variables will establish a positive correlation, implying that as the temperatures goes up, the permeability/absorbency also goes up. What’s more, if the temperatures rise above the membrane temperature, the cell membrane of the beetroot cell will be damaged. This implies that as the temperature goes up, the extent of damage will increase too. Aims of the study: To investigate the impact of temperature of the permeability of a cell membrane Materials and methods Materials 1. Thermostatic water bath – this is chosen because its accuracy is high given that it maintains water at the required level of temperature. This leads to results that are more reliable. 2. Size 4 cork - this is used to ensure a straight cutting of beetroot. 3. Test tubes – each water bath should have a test tube 4. Beaker 5. Thermometer 6. Heat proof mat 7. Paper towel 8. Tripod 9. Colorimeter 10. Measuring cylinder 11. Ten Beetroot cores, cut with a size 4 cork borer 12. Marker pen 13. One tile 14. Kettle for boiling water 15. mounting needle or forceps 16. Methods 17. This study will apply a controlled experiment, which will be performed in the lab. This method is preferred because it can easily establish the cause and effect. For example, in the current study, it can be easy to establish the cause of cell permeability as well as the effect of temperature on permeability. This will be achieved by manipulating a particular variable while keeping the others constant. Furthermore, lab experiment allows for accurate control of variables. For example, a calorimeter can be used to accurately measure the heat of chemical reaction, in the present study. A sample of 10 beetroot cylinders will be randomly selected and repeated experiments performed on them. Since similar patters will be obtained from the series of experiments, the validity of the study is relatively high and generalisation of the findings is more sensible. 18. The experiment will require control of factors, which will be maintained at the same level from one group to another, including the advance treatment of the beetroot cores, the service area and volume of the beetroot, use of water baths in the pre-heating of water in the test tubes, the treatment of cores after heating, the volume of water in the test tubes, and the extent of time that the cores are kept in the water baths. To increase the reliability of the experiment, the water temperature can be maintained with water baths that are thermal-controlled, and by using more samples of beetroot. The following are the materials and equipment used in the study. 19. Procedures 20. Preparation of the experiment will involve use of a size 4 cork borer to slash bores of beetroot. The beetroot is then soaked in a beaker of distilled water and left for one night. The following day, a series of water baths are set up at a range of temperatures. In order to start investigations, 3 or 4 beetroot cores are obtained from the beaker. Each of the cores is slashed into small pieces of 2.5 cm. This is done until one core for each temperature is available. Consequently, the 2.5 cm pieces are placed into a test tube – this test tube must be filled with enough distilled water. 21. For easy identification, each of the test tubes representing each of the water bath temperature is labeled with the level of temperature that it holds. Then, 5 cm3 of distilled water is added to each of the test tubes. These test tubes are place in each water bath, and left for about 6 minutes to be in line with the temperature of the water bath. Then, the beetroot cores are removed from the water and staining done lightly on a paper. The tissue can be handled with mounted needles or forceps, but it is important to assess the level of harm this might cause to the cores. In each of the test tubes, a 2.5 cm beetroot is placed lightly to ensure that all the pigment is well-combined with the water, and after this is achieved, the beet roots are removed from the cores. 22. What follows is the description of the deepness of the color that is contained in each of the test tubes. This is made more visible by a white piece of card if placed behind the test tube. Then, the test tubes are sorted according to the level of the water bath’s temperature. The relationship between the temperature and the amount of pigment discharged from the beetroot is then explained. In this case, a calorimeter can be used to provide response to green or blue filter and to calculate the absorbance. Finally, the absorbance of each test tube is measured and plotted against the temperature in order to establish possible patterns in the findings. 23. Ethical considerations 24. In examination of the impact of temperature on permeability of a cell, the cells of animals or plants can be used to make similar findings. However, due to the ethical considerations that animals should only be used when there is no other option; beetroots have been used instead of animals. What’s more, the objectives of the experiment are highly achievable, in which case they are highly likely have some befits in the long-term. Also, the performance of the experiment will be done with conformity to the necessary safety requirements. 25. Data analysis 26. The results of the experiments will be imported into Minitab software for various data analysis. This data will be manipulated and restructured in order to discover different relationships and patterns that will help in making an appropriate conclusion. This will include creating and interpreting various numerical measures and graphs. The specific outputs that will be drawn from the software include Histograms, Charts, Scatterplots, Paired test, and regression analysis among others (Dytham, 1999). The following table shows possible results that will be analyses in Minitab. Possible output are also suggested. 27. Temperature 28. Observation 29. Colorimeter reading (%transmission of light) 30. (C) 31. 32. Sample A 33. Sample B 34. Sample C 35. Mean 36. 0 37. clear 38. 100 39. 97.2 40. 89 41. 97.1 42. 23 43. very pale pink 44. 93.8 45. 94 46. 95 47. 94 48. 43 49. very pale pink 50. 80.2 51. 78 52. 77.9 53. 79 54. 636 55. pink 56. 26.5 57. 28.9 58. 32 59. 29.3 60. 88 61. dark pink 62. 0.8 63. 0.8 64. 2 65. 0.9 66. 94 67. red 68. 0 69. 0.2 70. 0 71. 0 72. Table 1: results of the experiment 73. Timeframe 74. 1 June 2013 – 15 June 2013 75. Review of literature and gathering of preliminary study materials 76. June 16 2013 – 1 July 2013 77. Gathering of experiment materials 78. July 15th 2013 79. Conducting of the experiment 80. July 16th – July 30th 81. Compilation of the report 82. August 1st 83. Presentation of the report 84. 85. 86. 87. 88. 89. 90. 91. 92. 93. 94. 95. 96. 97. References 98. Alberts B, Johnson A, Lewis J, et al., 2002. Molecular Biology of the Cell (4th ed.). New York: Garland Science. 99. Aveyard, A.H., 2010, Doing a literature review in health and social care. A practical guide. Maidenhead: McGraw-Hill open university press 100. Borrelli, M. J., Wong, R. and Dewey, W., 1986. A direct correlation between hyperthermia- induced membrane blebbing and survival in synchronous G1 CHO cells. J. Cell Phys. 126, pp. 181-190. 101. Budin, I and Devaraj, NK. , 2011. Membrane Assembly Driven by a Biomimetic Coupling Reaction. Journal of the American Chemical Society. 134 (2), pp. 751–753. 102. Cravalho, E. G et al., 1992. Response of cells to supraphysiological temperatures: experimental measurements and kinetic models. In Electrical Trauma: The Pathophysiology, Manifestations and Clinical Management. Cambridge University Press, Cambridge, UK, pp. 281-300. 103. Cruzeiro H., Ipsen, J. and. Mouritsen, G., 1989. Intrinsic molecules in lipid membranes change the lipid-domain interfacial area: cholesterol at domain interfaces. Biochem. Biophys. Acta. 979, pp.166-176. 104. Dytham A.C., 1999, Choosing and using statistics: a biologist’s guide. Blackwell science: oxward 105. Gamper, D., 2009. Investigating Factors That Affect Cell Membrane Permeability. Summer Research Program for Science Teachers. (Online) Available from http://www.scienceteacherprogram.org/biology/DGamper09.html (Accessed 10 April 2013) 106. Henriques, F. C. Jr., 1947. Studies of thermal injury. V. The predictability and the significance of thermally induced rate processes leading to irreversible epidermal injury. Arch. Pathol, 43, pp. 489-502. 107. Imgrund, R., 2009. Factors Involving Proteins. New York: SAGE 108. Kanehisa, M. I., and T. Y. Tsong. 1978. Cluster model of lipid phase transitions with application to passive permeation of molecules and structure relaxations in lipid bilayers. J. Am. Chem. Soc. 100, pp. 424-432. 109. Leyko, W., and G. Bartosz., 1986. Membrane effects of ionizing radiation and hyperthermia. Int. J. Radiat. Biol. 49(5), pp. 743-770. 110. Moussa, N. A., Tell, E and Cravalho, E., 1979. Time progression of hemolysis of erythrocyte populations exposed to supraphysiological temperatures. J. Biomech. Eng. 101, pp. 213- 217. 111. Padanilam, J. et al., 1994. Effectiveness of Poloxamer 188 in arresting calcein leakage from thermally damaged isolated skeletal muscle cells. Ann. NY Acad. Sci. 720, pp. 111-123. Read More