Drafts

Bacteriophages also have another type of life cycle. It is called the lytic cycle, the purpose of this life cycle is to rapidly replicate, lyse the cell, then package it’s DNA to go to a bacteria and infect another one. The lytic cycle has the bacteriophage replicate it’s DNA rapidly while making packaging for that DNA. The packaging includes the head and tail of the bacteriophage, this will keep the DNA safe until it gets to the next bacteria that it will infect. It also has to package it in a way that the bacteriophage will be able to insert its DNA into the next bacterial cell that it infects. This is a very interesting process. The goal of this process is to make as many bacteriophages as possible, this will allow the bacteriophages to leave the host cell and infect a bunch of other host cells. There are many proteins that the bacteriophage carry with them to make packaging for their DNA. And like in the lysogenic life cycle some bacteriophage use the host proteins to create their own packaging. Bacteriophages really are interesting, they infect bacteria which can infect us. There are viruses for everything, they can infect everything in life. The only thing that can’t be infected by a virus is a virus itself, and that is only because they can’t replicate on their own.

Chemical transformation of E. Coli cells is the process by which bacterial cell asre made to take up desired plasmids. Chemical transformation is performed after TOPO cloning. Some preparatory steps include equilibrating the water bath to 42⁰C, warming selective plates at 72⁰C for 30 minutes and thawing vials of One Shot Top 10 E.Coli cells on ice. To start off, 2 microliters of TOPO cloning reaction products are added into vials, mixed gently and then incubated on ice for 5-30 minutes. The cells are heat shocked for 30 second at 42⁰C without shaking before being placed on ice for 2 minutes. 250 microliters of S.O.C. medium is added, the tube is capped tightly and then shook horizontally at 200RPM and 37⁰C for 1 hour. 

Bacteriophages might be one of the coolest things in all of the natural world. They are viruses that infect bacteria. Bacteriophages are the pictures you see that look like an alien spaceship landing on a cell surface. They send their DNA into the bacterial cell. They can go through 2 separate life cycles, lytic and lysogenic. The lysogenic life cycle of the bacteriophage is for replicating DNA as fast as possible within the cell. What this means is that the bacteriophage replicates it’s own genome within the host cell replication machinery. Other types of bacteriophages package their own replication machinery so they don’t have to borrow it from the host cell. I think that packaging their own replication machinery is significantly less common than borrowing it from the host cell. It is more efficient for the bacteriophage to borrow the replication machinery from the host because it doesn’t have to waste space carrying it to the next host cell if it is already there. Typically during the lysogenic cycle DNA from the bacteriophage is inserted in to the bacterial genome. This is sometimes how genes are passed from bacteria accidentally. It is a really interesting process and sometimes drug resistance accidentally gets passed from bacteria to bacteria.

This disease known as the African sleeping sickness seems to affect a majority of sub-Saharan African countries. Tsetse flies are the main cause of this transmission of infection to individuals living within these countries. I thought it was interesting that these flies only inhabit tropical Africa and are large biting parasites that live by feeding on the blood of animals. The blood of the animals is where initially the flies themselves are infected. I also think that because of the scarce and in availability to the treatment of this disease is how the spread of the disease continues to infect so many individuals. There has been a reduced number of new cases since 2009 which used to be 10,000, meaning that the research and discovery of treatments and medications is helping the countries infected. The goal of this renovated medication is to stop the disease at an earlier stage and prevent the widespread of infection. Because the disease can be fatal, I think that with an earlier treatment could help prevent the loss of many human lives as well.  

Oviposition preferences of the female azuki bean weevil determine the success of the offspring. The choice of the female determines the food source for the larvae, and because the bean on which it was laid will be the only food source for its entire development, the right decision on the female's part is crucial. After pupation, the beetle does not consume food or water and will seek to reproduce. In experiments that test for preferences, it is found that the azuki bean weevil has a high preference for the cowpea legume out of five beans- cowpea, mung bean, azuki bean, kidney bean, and soybean. The number of eggs per cowpea when the female was given a choice revealed roughly 6 eggs per female on the cowpea, and zero eggs per female on the kidney bean. Furthermore, the developmental time and the rate of adult emergence was reduced for the eggs laid on cowpeas, suggesting that the choice was beneficial to offspring. Further research on other species suggests that certain legumes produce a higher concentration of phenol, found to be toxic to certain species, thus leading to avoidance on those legumes.

For the treatment regarding the three mi-RNAs in ovarian cancer associated fibroblasts, I tried to figure out downstream targets by which these epigenetic changes of mi-RNAs in CAFs induce epithelial to mesenchymal transition of cancer cells. I proposed a few possible pathways, although these are likely far more complicated, and there are likely many more that I did not cover. What makes mi-RNAs interesting is that because they are so short, approximately 20 bases in length, they often have binding specificity for thousands of genes. This means that treatment involving mi-RNAs usually evolves from an observational study about mi-RNA dysregulation, and the reversal of that phenotype having anti-cancer effects. With mi-RNA treatment, it is often the case that it is discovered that reversal of the phenotype is effective in treating cancer, without figuring out exactly why, because the possibilities are so vast.

0.75 mL of acetic acid and 1.20 mL of isopentyl alcohol were added to a round bottom flask. Four drops of concentrated sulfuric acid was carefully added to the flask before being refluxed in an air condensing distillation apparatus. The solution was boiled for 15 minutes and then the organic layer of the collected liquid was poured back into the round bottom flask and boiled for another 15 minutes. The process was repeated one more time for a total reflux time of 45 minutes. The solution was then cooled to room temperature and poured into a centrifuge tube with 1 mL of water. The solution was mixed and the bottom layer was removed to waste. 1 mL of sodium bicarbonate was used to rinse the solution and the aqueous layer was removed again. Then 1 mL of sodium chloride was used to rinse and the aqueous layer was again removed. Calcium chloride spheres were added to the solution and then the dry solution was moved into a tared vial were the odor was observed. Lastly, an IR test was performed to determine the purity and identity of the isopentyl acetate.

The aim of the project is to gain more insight into the function of the alpha-4 GABAA receptor by the deep examination of subunit expression in the brain and spinal cord of zebrafish. The in-situ hybridization procedure done previously (Monesson-Olson et al., 2018), allowed the visualization of six subunit expression patterns, including that of alpha-4. This gave a rough picture of subunit function. Since the mutation screen revealed interesting alpha-4 subunit activity, further analysis is required. Conducting this fluorescent in-situ hybridization procedure will allow me to identify not just where the alpha-4 subunit is expressed but also what structures it interacts with. Knowing what the alpha-4 subunit interacts with will aid in predicting its function. By extension, this will provide a framework for developing treatments that can suppress the effects of related neurological disorders like epilepsy.

Monticello is located just outside of Charlottesville, Virginia, which has a four-season humid subtropical climate with all months being well-watered, though the period from May to September is the wettest. Winters are somewhat cool but mild, and Summers are hot and humid. The site was excavated using test units of equal sizes to keep the sampling strategy consistent. Each test unit was excavated stratigraphically using shovels and hand trowels. Stratigraphic layers were differentiated by soil color and texture. All of the sediment removed from the test units was sifted through a screen, and all artifacts were collected and labeled according to unit and layer. Summertime temperatures are high in the region, with indoor temperatures of around 100 °F. Jefferson himself is known to have been interested in ancient temperature-control techniques such as ground-cooled air and heated floors. Monticello's large central hall and aligned windows were designed to allow a cooling air-current to pass through the house.

Many of the countries where most of the cases occur; put these treatments out of reach for many patients because of its complexity. The new drug, fexindazole, is a much simpler treatment in which it can be taken as a once-a-day pill for 10 days without having to be hospitalized. After the DNDi (Drugs for Neglected Diseases Initiative), a nonprofit organization, teamed up with drug maker Sanofi, they could test the drug on patients and apply for an EMA recommendation for the drug itself. The EMA has just recently approved of the drug and have expressed a “positive opinion” for the fexindazole, opening opportunities for patients to receive treatment earlier and help slow the spread of the disease as well.