Drafts

Surface and interior images of fixed NIH 3T3 cells treated with fluorescently labeled transferrin and lysotracker were taken at 100X magnification at three separate time points (0 min. incubation, 30 min. incubation, 90 min. incubation) in order to capture endosome migration over time (Figure 1). At t=0 min., it is clear that there is an abundance of transferrin molecules at the surface of the cell (Figure 1A), resulting in a relative high average total fluorescence intensity compared to the interior of the cell (2.5, 2.1 respectively) (Figure 2). At t=30 min. the average total fluorescence intensity of the surface and interior of the cell is relatively similar (3.3, 3.2, respectively) (Figure 2), At t= 90 min. there is a relatively higher average total fluorescence intensity at the interior of the cell compared to the surface of the cell (4.1, 3.3, respectively) (Figure 2).  Additionally, it can be visually noted that there is a significantly larger presence of transferrin molecules in the interior image than the surface image (Figure 1H, 1G, respectively). Lysozymes are unable to be visualized due to overpowering green stain in composite images. 3D projections of the cells were unable to be created due to immense technical difficulties.

The process by which cells intake extracellular material from the environment is known as endocytosis, and can occur in both a receptor-mediated and non-specific manner. This laboratory exercise investigates both of these endpcytotic processes over time using fluorescence microscopy to observe endosome migration in fibroblast (NIH 3T3) and epithelial (LLC Pk1) cells treated with transferrin and TMR-dextran. Additionally, effects of nocodazole in LLC Pk1 cells was studied to explore the efficacy of vesicle motility along microtubules in cells with depolymerized tubulin.

To a 10 mL round bottom flask sodium hydroxide (0.064 g, 1.60 mmol), 2-naphthol (0.200 g, 1.39 mmol), and a few boiling stones were added. Next, absolute ethanol (3 mL) was added and the air condenser was attached. The mixture was refluxed for 25 min while the sand bath was at 30% heat and cooled for 3 min. Then, n-butyl iodide (0.200 mL) was added to the mixture through the top of the condenser and reflux continued for 1 h. After reflux was complete, the reaction mixture was poured over ~10 g of ice in a beaker. The round bottom flask was then rinsed with cold water (2 x 1.0 mL) and this was transferred to the reaction mixture. The reaction mixture was stirred with a glass rod until 95% of the ice melted and the final product (0.036 g, 13.6% yield) was collected via suction filtration after being rinsed with ice water. A small portion of the product was then analyzed using thin layer chromatography and different solvent systems of ethyl acetate and hexanes were experimented with until a decent separation between starting material and product was seen.

Females are attracted to territories with more vegetation because it attracts more prey for them and serves as a water supply which they are able to retain well due to the reduced number of sweat glands. The females will go to the male with the most vegetation in his territory, eat and mate. The female American Rhingon mates once a year for the last three of her five-year life span. The females reach sexual maturity after two years while the males reach sexual maturity after only one, however both male and female are abandoned by their mother after six months of nursing. Mating season is during the months of April and May, this is because they estivate, deep in the sand, during the months of June to August due to the unbearable temperatures and no rainfall. During this period of dormancy, they develop their young who take the full three months to develop. Their core temperature cools due to the burrow which slows their metabolism enough to keep the mother alive and to nourish the embryo. They return from their dormancy period once the young have developed and temperatures have dropped, their young being altricial as their dormancy does not aid the full development but reach independency after six months.

BRM270 when combined with the use of miRNA-128, which acts as a tumor suppressor downregulated in lung cancer would be a great way to combat chemoresistance of NSCLC [19]. Instead of using the virus we could use this compound combined with miRNA-128 to combat the resistance developing of gefitinib. Increases sensitivity of TKI. BRM270 has a low toxicity and targets the NF-kB signaling pathway, induces cell cycle arrest and apoptosis. This is combined with the downregulation of marker CD133. Since we want to suppress Vimentin in our initial method using BRM270 will be another way to suppress Vimentin and induce E-cadherin which is good because then this will mean we are progressing to inhibiting metastatic progression.

BRM270 is made up of 7 distinct types of plants known as Saururus chinensis,, Aloe vera, Arnebia euchroma,Citrus unshiu Markovich, Portulaca oleracea, Scutellaria baicalensis, and P.vulgaris var. Lilacina. Studies have shown that this substance plays a role in inhibiting tumor cell proliferation [19]. Some of these extract plants are known to be involved in anti-inflammatory activity like P.vulgaris var. Lilacina [20] BRM270 has been illustrated to suppress formation of tumors and does this by negatively regulating nuclear factor-kB signaling in drug resistant cancer stem cells. A study was conducted in which lung adenocarcinoma cells migration and invasion to other areas as well as chemoresistant derivatives was inhibited by treating it with BRM270.

What were the authors trying to test, and what predictions did they make?

    The authors were assessing the genetic diversity and relatedness in the critically endangered species hawk Buteu ridgwayi. The island population of birds is expected to exhibit a lower genetic variation than mainland populations. This is because there is reduced gene flow, smaller population sizes and increased genetic drift due to isolation. The island birds also likely suffer from inbreeding depression.

What, exactly, did the authors do?

    The authors collected samples, and monitored nests and breeding pairs over four years. Blood was collected from 149 Ridgway’s Hawks. Fourteen microsatellite loci were tested to examine allelic variation. The program Bottleneck was used to test for the presence of a genetic bottleneck. Alrequin was used to calculate the observed heterozygosity, expected heterozygosity, and number of alleles. The relatedness was determined used a program called Kin Group.

What did the authors find (i.e, what were their data)?

    The allele frequencies ranged from 0.0069 to 0.4960. The observed heterozygosity across loci ranged from 0.521 to 0.884. Relatedness coefficients of the seven sampled breeding

pairs ranged from -0.09 to 0.55.  There was no structure present, all of the samples came from one genetic population. A significant excess of heterozygotes was found.

How did the authors interpret their findings?

    The results from the study show that the population underwent a recent genetic bottleneck, had a high level of heterozygosity, and occurred inbreeding. The significant excess of heterozygotes indicates a recent genetic bottleneck. The relatedness analysis verified the presence of inbreeding. Island avian populations are of great concern, because they have a higher risk of extinction than mainland animals. These island birds are more susceptible to extinction primarily due to human caused habitat loss and introduced species.

The target product of hydrobenzoin was obtained thorugh a reduction reaction of benzoin using sodium borohydride as the reducer and resulted in a 55% yield.  Recrystallization with acetone as a solvent was used to separate the hydrobenzoin product from remaining impurities.  The presence of hydrobenzoin was confirmed from the thin layer chromatography performed. 

Regarding abortion, however, Warren argues that the above cases do not apply to fetuses, even if there are many who view fetuses as valuable. Warren argues that so long that the fetus remains unborn, its survival is dependent on the mother, and thus infringes on her rights to freedom, happiness, and self-determination. Warren argues that a woman’s rights override those of who would like to see the infant preserved. She compares this situation to that of a man defending himself against a wild animal; he has the right to kill it in defense, even if there are those that wish for to see the animal live. Warren continues by stating that when the fetus is born, its preservation no longer violates any of its mother’s rights even if she wants to destroy it, because she is free to put it up for adoption. As such, the moment of birth marks the end of the mother’s right to decide its fate (Warren, 2).

Therefore having a lower recruitment of neutrophils in the lung could result in a decrease of lung metastasis with TLR3 already inhibited. This then suggest that TLR3 is also involved in promoting pre-metastatic niche creation by recruiting neutrophils to the lung [7]. When TLR3 is present in host stromal cells than chemokine production is high and this then means that neutrophils have an easy way to mobilize and be recruited which we already know are known to contain Bv8 and this will then mean that transportation of tumor cells and facilitation is increased as well [7]