Hyperglycaemia and oxidative stress upregulate HSP60 & HSP70 expression in HeLa cells

Heat Shock Proteins 60 & 70 (HSP60 & HSP70) are intracellular protein that has been shown to be present at elevated levels in systemic circulation in Type 2 Diabetes mellitus (T2DM) patients. Conditions that lead to its secretion, and the mechanism of its translocation from cells, have not yet been defined. The aim of this study was to determine if specific cell stressors associated with T2DM, namely hyperglycaemia and oxidative stress, result in the upregulation of HSP60 in human cells in vitro. Human HeLa cells were grown in media supplemented with 100 mM glucose, 200 μM hydrogen peroxide (H₂O₂), and 50 μM sodium azide. Initially, the effect of these treatments on cell growth rate was examined, with each treatment significantly inhibiting growth rate. LDH and MTT assays were also used to successfully demonstrate that these treatments do not significantly increase cell lysis, but do significantly impair mitochondrial dehydrogenase activity. To confirm this mitochondria specific form of inhibition, DCFDA assay were used to investigate any increases in intracellular reactive oxygen species (ROS) generation. All three treatments resulted in significantly increased ROS generation, with greater ROS production occurring with a greater exposure time. Interestingly, when the protein levels of HSP60 and HSP70 were measured after 3 and 7 days of exposure of the HeLa cells to 100 mM glucose, 200 μM H₂O₂, and 50 μM sodium azide significant induction of these two molecular stress proteins were observed ranging from 2.43-5.08 fold compared to untreated control cells

Summary of Middleton vs. Warden, 120 Nev. Adv. Rep. 74

The Nevada Supreme Court removed Middleton’s appointed appellate counsel due to substandard representation. On initial review, the Nevada Supreme Court ordered Middleton\u27s counsel to submit an amended brief, limited to 80 pages. Counsel\u27s amended brief was simply the original brief with the final few pages removed so as to meet the 80-page requirement. Counsel had repeatedly violated court orders, and the work product he ultimately submitted was unacceptable for representation of a client who was facing a death sentence

Summary of MGM Mirage v. Cotton, 121 Nev. Adv. Op. 39

When MGM Mirage employee Brenda Cotton walked through her employer’s parking lot ten minutes before her shift, she tripped over a parking lot curb and injured herself. Cotton sustained an ankle fracture and ligament tear. Deciding that Cotton had failed to prove that her injury arose out of her course of employment, MGM denied her workers’ compensation claim. MGM’s decision was upheld by a hearing officer, because the injury did not occur during working hours. The hearing officer’s decision was reversed on appeal. MGM’s petition for review was denied by the district court

The Characterization of BECy-PES Polymer Blends

The bisphenol E cyanate ester (BECy) and Poly(ether sulfone) (PES) blend is a very unique system with a promising future in the aerospace and military industries; for it combines the positive attributes of both polymers. BECy is a thermoset that has very high strength; performs well in high temperature environments; and it is a flowable liquid at room temperature, which allows for easy processing. PES is a thermoplastic that is a good toughening agent with good thermal attributes. However, there is much about the BECy-PES blend that has yet to be discovered. Thus, a study was performed in collaboration between Dr. Samy Madbouly and Dr. Nicola Bowler that sought to further characterize the BECy-PES blend in a non-cured state. Different compositions of the blend were created and characterized with rheology; Differential Scanning Calorimetry (DSC); and Thermogravimetric Analysis (TGA). As a result of the rheology study, blend samples with high concentrations of PES (above 40 wt%) had higher viscosities than samples with lower concentrations of this polymer. The DSC data indicated that samples with higher concentrations of PES also had higher glass transition temperatures than the samples with lower concentrations. Last, the TGA data suggested that the blend undergoes curing at 200°C

Summary of United Insurance Company of America vs. Chapman, 120 Nev. Adv. Rep. 83

The Nevada Supreme Court ruled that the prejudgment interest awarded to Chapman should have been calculated pursuant to the specific interest statute, rather than the general interest statute, because the special interest statute was in effect when the judgment was entered. The district court had erred by awarding the attorney fees to Chapman, because United’s claim was brought on reasonable ground. The award of postjudgment interest awarded to Chapman was also reversed

Long Term Effects of Chemotherapy on Cognition, Preventative Potential of Antidepressants

Each year, over 1.6 million people in the U.S. alone will be diagnosed with some form of cancer. With advances in treatment, survival rates have risen to nearly 65%. While remission and survival are the ultimate goals of treatment, it has become clear that many cancer survivors (estimates range from 15% to 70%) treated with chemotherapy experience significant, long-lasting cognitive impairment. This chemotherapy associated cognitive impairment is often called Chemo Fog or Chemo Brain. For some, the effects are mild, such as having difficulty with focusing, concentrating, and speed of processing. For others, the cognitive impairments can be significant and may dramatically alter their day-to-day lifestyles, even preventing some from ever returning to work. Growing evidence suggests these impairments are sometimes permanent. The present study attempts to identify the underlying mechanisms of Chemo Fog that contribute to impairment in learning, memory and processing speed, associated with chemotherapy administration. This study also investigates a potential antidepressant treatment that, when co-administered, may reduce the effects of chemotherapy drugs. Two proposed explanations as to why such changes follow chemotherapy treatment are; a reduction in the proliferation of new brain cells, and a decrease in the myelination of axons. A B6 strain of mice was used as a model for this investigation. Animals were administered the chemotherapy agent 5-Fluorouracil (5-FU) and five a known neuroprotectant (fluoxetine) over a period of 5 days. Neural tissue was examined to assess the effects of the treatment at 1 day, 14 days and 6 months post-treatment. A chi squared test was used to analyze data from day 1 and day 14 tissue. A Oneway Anova test was used to analyze data from 6-month tissue. Analysis revealed a significant reduction in myelin at 1 and 14 days but not 6 months for individuals that received only 5-FU as compared to controls. Analysis also showed that animals treated with fluoxetine and 5-FU had myelin that was consistent with control animals for short-term time points (1+14 days) suggesting myelin preservation. Statistical tests showed no myelin protection for the animals treated with fluoxetine at the 6-month time point as their tissue was not statistically different from animals receiving 5-FU alone