Expression of TRPV5 in Astrocytes: Implications for Ischemic Stroke
Elevation of intracellular calcium secondary to increased calcium influx along with increased gliosis are implicated in the pathogenesis of focal ischemic stroke. In astrocytes, which play a major role in maintaining homeostasis in brain ischemia, the identities of the ion channels responsible for increased calcium influx during ischemia is relatively unknown although several Ca2+-permeable transient receptor potential (TRP) channels have been identified to have contributing roles. The transient receptor potential vanilloid 5 (TRPV5) channel is a Ca2+-permeable cationic channel expressed primarily in kidney epithelial cells and at low levels in the brain, but the exact localization and role this channel plays in the brain has not been explored. To investigate the possible role TRPV5 plays in astrocytic calcium influx in ischemia, we examined the functional expression of TRPV5 in astrocytes subjected to hypoxia-ischemia in vitro and in rat models of ischemic stroke in vivo. We hypothesize that TRPV5 contributes to increased calcium influx in ischemia. By treating astrocytes with culture conditions without glucose and with low oxygen levels, we found that TRPV5 is upregulated with increasing durations of simulated hypoxia-ischemia in vitro. Similarly, rat models of ischemic stroke with middle cerebral artery occlusion also show TRPV5 upregulation in reactive astrocytes, suggesting a possible role of TRPV5 in reactive gliosis in vivo. Microfluorimetric intracellular calcium imaging using Fura-2 on primary cultured astrocytes show a voltage-independent increase in astrocytic calcium influx after hypoxia-ischemia in vitro that is selective for extracellular Ca2+ concentration and is reduced by inhibition of TRPV5 with ruthenium red. Electrophysiology measurements using the whole-cell patch clamp technique on primary cultured astrocytes reveal a non-selective cation current similar to that of TRPV5 that is inhibited by Mg2+, another inhibitor of TRPV5. Preliminary results on astrocyte cell viability during hypoxia-ischemia with TRPV5 inhibition by ruthenium red also suggest that inhibition of TRPV5 could enhance astrocyte survival and reactive gliosis in vitro, indicating a beneficial role in blocking non-selective Ca2+ entry via TRPV5 into astrocytes. Since TRPV5 is highly selective for Ca2+ and an important channel for Ca2+ absorption in various epithelial cells, TRPV5 upregulation may contribute significantly to elevated Ca2+ influx in astrocytes in hypoxia-ischemia. Also, Ca2+ influx has been demonstrated to play a crucial role in reactive gliosis, further suggesting that TRPV5 upregulation is involved in reactive gliosis. We propose that TRPV5 is involved in ischemia-induced calcium influx in astrocytes, and might participate in the pathogenesis of focal ischemic stroke.
What really attracts boys to girls
The purpose of this research was to challenge misconceptions about what attracts teenaged boys to girls. It has been observed that some teenaged girls hold stereotypes about boys and believe boys are only attracted to superficial qualities like beautiful hair. The aim is to investigate differences between what girls believe boys find attractive about them and what boys actually find attractive about girls. Hypothesis: There is a significant difference between what teenaged boys find attractive in girls and what teenaged girls think boys find attractive. Procedure Background reading was done to discover which characteristics are commonly used to measure attractiveness among teenagers. A survey questionnaire was used for gathering data. To gather information for the construction of the questionnaire, 4 focus groups were interviewed. The groups were asked to determine indicators of attraction and repulsion. Two separate questionnaires were designed (one for boys and one for girls), based on focus group information, with similar questions which could be used to compare results. Questions were related to personality and looks. The research was representative as 399 teenagers participated in the final survey. Results were collated in Excel tables. Percentages were calculated by using a scientific calculator and Excel. A bivariate analysis was created to examine the main question: ranking qualities that attract boys to girls. A comparison was made between boys’ and girls’ opinions from different age categories. Mean averages were calculated for this question. Other questions were analysed using mode averages translated into percentage of each gender. Graphs were designed using the values calculated. Observations/Data/Results 72% of boys prefer a girl with average looks and an awesome personality but only 37% of girls believed that to be true. The average boys from 12-14, 15-17 and 18-20 year old categories agree that confidence, humour, a nice body, politeness and intelligence are the five most attractive qualities in a girl they would consider dating. The order of these five change across the age groups but are constantly the top five, however confidence and humour always trump a “nice body”. The average 12-14 and 15-17 year old girl overrates a nice body and underrates politeness and intelligence but confidence and humour feature in their top five. The average 18-20 year old girl agrees with the boys priorities. When defining what makes a girl beautiful, 68% of boys highlight personality traits and 59% of boys mentioned physical features. When noting what repulses boys about pretty girls, more than 50% of both sexes agree that smelling bad was most off putting. 52% of boys mentioned taking drugs as unattractive yet only 23% of girls recognized this as repulsive. Conclusions The data supports the hypothesis. Confidence and humour are more important to the average boy than a nice body across all age groups. The younger the girl, the less she understands what attracts boys. 63% of teenaged girls believe that looks are more important to boys than personality. This is clearly not true. 72% of boys value a great personality more highly than good looks.
Totarol
Research Question: To what extent does totarol show antibiotic potency against significant strains of Staphylococcus aureus causing skin and soft tissue infections in New Zealand, compared with commonly prescribed drugs used to treat the specific strain of Staphylococcus aureus being tested? Purpose of research: This essay will investigate the antibacterial potency of totarol against significant strains of Staphylococcus aureus (S.aureus) causing skin and soft tissue infections (SSTIs) in New Zealand. Only a small amount of research on totarol has been conducted. They all suggest totarol possesses antibiotic potency against various species of bacteria. The mode of action of totarol is currently unknown. Procedure 1: The totarol I ordered from Mende Biotech Ltd arrived in two forms; a powder called Totarol™ and a viscous brown liquid called Liquid K7 (LK7) in which the Totarol™ powder was dissolved in mostly sunflower oil. The investigation took place in two stages. In the first stage, the MIC value of the LK7 against reference strain S.aureus 29213 was determined by following the CLSI methodology for MIC testing (M07-A9 Clinical and Laboratory Standards institute). The MIC test was also conducted to identify whether any major ingredients in the LK7 possessed significant antibacterial potency. MIC values where compared with that of flucloxacillin. Data from stage 1 testing: LK7 had an MIC value of 1µg/mL, which was very similar to flucloxacillin’s MIC value of 0.5µg/mL. No other major ingredients in LK7 showed antibacterial potency. Totarol™’s antibacterial activity could not be accurately measured, due to the powder resisting even mixing. Procedure 2: In the second stage, disc diffusion tests were conducted against various S.aureus clinical isolates obtained from SSTIs in the Waikato community. The discs that were placed for each clinical isolate included LK7, cefoxitin, fusidic acid, mupirocin and erythromycin discs. Data from stage 2: 75% of LK7 discs produced double zones of inhibition. I hypothesized that this was due to two active ingredients found in the LK7. I predicted the one that produced larger zones of inhibition to be Totarol™. The other more stable ingredient producing the inner zones of inhibition is unknown. Conclusion: I proposed a breakpoint of outer zone sizes that were ≥ 15 mm in diameter to signify that that particular clinical isolate was ‘susceptible’ to LK7. From this breakpoint, LK7 and fusidic acid both had the same number of clinical isolates that were classified as ‘susceptible’. LK7 was the median of the number of susceptible clinical isolates. This data answered my research question; totarol in the LK7 form specifically, would be just as effective in treating SSTIs caused by S.aureus, as even the most commonly prescribed antistaphylococcal drugs currently being used.
Carbon Nanostructures Via Dry Fce Exposed to High Temperature
This science project is designed to answer a question of whether or not a chemical reaction is needed to produce industrial quantities of carbon nanostructures by exposing dry ice to a high temperature that is at least 3100°C. A small carbon arc furnace powered by an electric welder is used to produce the high temperature. During control runs, the carbon arc furnace is energized for a predetermined time, after which the carbon arc furnace is de-energized and any carbon particles within the furnace are collected. During carbon nanostructures synthesis runs, dry ice is placed within the carbon arc furnace. The carbon arc furnace is energized and the dry ice is consumed for the predetermined time. Carbon nanostructures synthesized during the synthesis runs are collected once the carbon arc furnace is de-energized and allowed to cool. The volume of the carbon particles collected during the control runs is compared to the volume of the carbon nanostructures produced by the synthesis runs. This science project has discovered that on average at least 16 times more carbon nanostructures are produced during synthesis runs consuming dry ice as opposed to the control runs. Moreover, the synthesis runs did not rely on chemical reactions. Further still, samples of the synthesized carbon nanostructures were imaged using a transmission electron microscope (TEM). The TEM images clearly show high-quality carbon nanostructures that include carbon nanotubes, faceted carbon nanospheres, and the super-material graphene.