Ethanol Induced Stress In Yeast

"Ethanol Precipitation of DNA and RNA: How it works." Bitesize Bio. N.p., n.d. Web. 20 Feb.

The expression of lanes 3,4,6,7,8 and 9 were expected but the expression of lane 5 was not expected. The reason lanes 3,4,6,7,8 were expected is that lanes 4,5,7,8 contain genomic DNA, which should contain multiple genes including the genes for Heat shock resistants. Also the genomic DNA in lane 4 and 8 under went heat shock condition, which led to the expression and amplification of Heat shock resistant proteins. This is also the case for lane 6, which contains heatshock cDNA. Since it contains cDNA that underwent heat shock condtion, that means the gene is being express and amplified in order to resist those heated condotions.

One hypothesis was that the potential mutagens, Diet Coke and ThermaFlu, were in fact mutagens. With that in mind, the reason for carrying on this experiment was to test and see if our hypothesis was correct. As the experimentation proceeded, data showed that Diet Coke was the best potential mutagen, and was chosen to continue testing. With this substance being the focal point of the second experiment, the goal was to test different amounts of Diet Coke with the TA 1535 strain of S. typhimurium. Hypothesis was that as the concentration of Diet Coke was increased, colony growth would raise as well.

The Alcohol in the agar interferes with the DNA synthesis of Gram-negative organisms which inhibits growth.

Following this initial study, our laboratory examined consumption behavior in mice that have a genetic knockout of the FABP5 gene. Similarly, both male and females underwent a limited access two bottle choice paradigm. The genetically knocked out (KO) animals were compared to wildtype. As originally hypothesized, the genetically modified animals consumed significantly more ethanol compared to wildtype animals. Conversely, following 30 minutes restrain stress, the FABP5 KO animals had a reduced corticosterone response compare to the wildtype animals. These results contradicted our previous study, using the pharmacological inhibition of

Three wells where eliminated from the experiment, because a vehicle control was not needed for this experiment. Images where then taken of the 9 wells, with the use of phase-contrast microscope, these photos were labeled (T0). After the images where obtained, the well-plate was placed back into the tissue culture hood, and the wells where treated with different treatments, so the effects could be observed. In row “A” contained the HepG2 cells by themselves, no treatment was added. Row “B” contained the HepG2 cells and 4.5µl of Ceramide, which was labeled as the positive control, and finally in row “C” the wells contained our HepG2 cells and the compound Epigallocatechin Gallate. The well was then placed back into the cell culture incubator for 24 hours. Following the incubation, photos where taken using the same phase-contrast microscope and the same area of each scratch, these photos are labeled (Tf). The before and after photos of the scratch are then used to figure out the percent wound closure, which determines the cell migration. In order to calculate the percent closure, the following equation was used; Percent closure=width of Tf box/width of the T0

On the other hand, the investigation of the apolipoprotein genome, the identification of specific loci, the role of alcohol dehydrogenase

The sun is a source of UV light that is very hard for most people to escape. It affects our cells in various ways and sometimes causes cell death. In this experiment, to inspect the damage done by UV irradiation on the genetic composition of Saccharomyces cerevisiae also known as common Baker’s yeast. The strains of this yeast used were the cells that were mutated and the TRP1 gene was inactive. In this strain, the cells would not be able to produce tryptophan, which they need in order to grow. Phenotypic reversion of this gene was examined by first spreading the cells on SD medium without tryptophan and a complete SC medium with tryptophan. The yeast on the SC medium while they SD mostly died on the plate. However, in part two of the experiment whenever these plates were exposed to UV light the TRP1 gene was reactivated and growth occurred. On the SC plates as time went on there were less colonies formed. In turn, on the SD plates the pattern of growth followed a bell shaped curve; there were more colonies after the plate was exposed to half the time maximum time then when the plate was exposed to the maximum time. These results indicate that mutagenesis of Saccharomyces cerevisiae did in fact occur.

Alcohol has been shown to alter normal brain development by interfering with normal cell processes responsible for cell division and gene expression. Guerri and Renau-Piqueras (1997) explained that alcohol affects cell division by inhibiting cytoskeleton activity. This increases the cells in the G0 and G1 phase, which reduces the number of mitotic cell undergoing cell division. A later study by Anthony, Zhou, Ogawa, Goodlett, and Ruiz (2008) found similar findings in the reduction of mitotic cells, but through induced apoptosis. They used mouse dorsal root ganglion stem cells to research the affect of PAE on the G1 and S phase of the cell cycle. Cell cultures were either exposure to 200 mg/dl or 400 mg/dl of alcohol, excluding the control group. These researchers found that alcohol exposure increased the levels of cyclin D1, cyclin D2, and E2F1, each of which play an important role in the regulation of the G1 and S phase of the cell cycle as well as subsequent cell division. Over expression of these proteins leads to decreased regulation in the cell cycle, altered DNA synthesis, increased DNA fragmentations, and ultimately apoptosis. This supports their findings of reduced cell numbers in the cultures exposed to alcohol.

Approximately 88,000 people die annually due to alcohol abuse. There is currently no medication intended to suppress the desire for alcohol; however, it may become a reality in the very near future. In the last year scientists have discovered that the Beta-klotho gene, originally thought to only regulate the progression of the FGFR4 protein in hepatocytes, may have multiple uses. Although it’s not extremely common in people, the gene may have the ability to be used in a drug to help repress the urge to drink alcohol. Scientists speculate the gene may also be the key to regulating obesity and heart disease.In other words, the Beta-koltho gene could potentially solve a few of society’s biggest

A great proportion of alcohol metabolism occurs in the liver; this is also the primary location of the production of glucose from the breakdown of glycogen (glycogenolysis) or from the breakdown of non-carbohydrate sources - amino acids, pyruvate, glycerol (gluconeogenesis). Alcohol is detoxified and removed from the blood through a process called oxidation. Oxidation prevents the alcohol from accumulating and destroying cells and organs. Alcohol is metabolised by an enzyme in the liver cells known as alcohol dehydrogenase (ADH). ADH and ALDH oxidise ethanol into acetate. First ADH breaks down alcohol into acetaldehyde and then another enzyme, aldehyde dehydrogenase (ALDH) rapidly breaks down acetaldehyde into acetate. Acetate is further metabolised and eventually leaves the body as carbon dioxide and water. This oxidation process consumes 2 NAD+ generating 2 NADH. The high concentration of excess NADH produced inhibits gluconeogenesis by preventing the oxidation of lactate to pyruvate. Instead, it causes the reverse reaction to occur, resulting in lactate to accumulate – this can result in acidosis (lactic acid build-up) and hypoglycaemia from the lack of glucose produced. Pyruvate is the first designated substrate of the gluconeogenesis process which can then be used to synthesise glucose, hence why when alcohol is inhibited, gluconeogenesis doesn’t

Ethanol is a non-polar and uncharged alcohol, when at high concentrations, will destroy the lipid bilayer barrier of membranes and increase ion

). The low tolerance might be due to the concomitant acetaldehyde and acetate formation during ethanol reassimilation in these xylose-fermenting yeasts. Acetaldehyde is very toxic to the cell, and concentration above 0.5 mM inhibits all cellular activity (Liberthal et al., 1979). Lucas and van Uden investigated the effects of temperature on ethanol tolerance and thermal death of xylose-fermenting yeast and determined that it was more tolerant of ethanol at lower temperatures (Lucas and Van Uden, 1985). Effect of ethanol on metabolic rate has been examined with ethanol added exogenously. Both Lucas and van Uden and du Preez et al. placed cells into media containing different concentrations of ethanol and measured the specific growth rate (Preez

If I was to do an experiment like this again I could use a different variable instead of alcohols. Instead I could investigate a chain of alkanes or alkenes, “in order to test alkenes I would have to look into the association double carbon bonds have with one

Because alcohol is not found easily in nature, genetic mechanisms to protect against excessive consumption may not have evolved in humans as they frequently have for protection against natural