Monday, April 6, 2020
On August 30, 1831, is the most exciting news for Charles Darwin. He got the offer of a position on the H.M. S. Beagle. This offer was from his guidance, Henslow, at Cambridge. Henslow had been welcome to be the natural scientist for the ship, but he had turned down the opportunity. Fitzroy is a twenty-six-year-old male, and he was the captained at the time. He was anxious to have the brotherhood of somebody who was of his social class. Majority of the experienced naturalists had turned out to be occupied or reluctant, so Henslow gave the opportunity to Charles Darwin who has no experience. Sadly, there was an obstacle to be crossed before he is allowed to take off on his journey. He was required to get his blessings from his father, Robert, but the dad had enough of Charless laziness and attitude. Charles left the town, so he can talk to his uncle, Josiah, about his problems. We will write a custom essay sample on On August 30, 1831, is the mos or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page Josiah concurred with his nephew that this was an ideal opportunity. He thinks that there was absolutely no motivation to believe that the voyage would interfere with his profession when he returned. Later, the uncle composed a letter to Robert listing the reasons why the voyage would be useful for Charles. He came back to his hometown to find that his father had been persuaded. If Josiah was not there for the help, Charles would never get the experience to travel. Charles was happy. The ship was originally planned to depart in two or three weeks, but as he was getting ready, he got bad news. It has been said that there was a miscommunication and Fitzroy had just guaranteed the situation to a companion. Charles would only get the position if the companion cannot or declined the request. Regardless of the misfortune, he raced down to London to meet the captain for a meeting. Fitzroy appeared to be unpredictable. A few days later, Fitzroys companion gave up the position, so Charles has taken the position. He additionally discovered that the journey will probably be somewhat three years than two. The ship, H.M.S. Beagle, was delayed so many times even though it is readied, but poor climate postponed the crew much more. On December 10, they finally set sail, yet were soon turned back by storm winds that caused trouble and left Charles wretchedly disgusted. He was not happy about it. On December 21, couple days before Christmas, they had what resembled consummate climate and attempted once more. Fitzroy had an awful start, so he quickly controlled the ship on solid land, however luckily nothing was harmed. When Charles woke up after his first nights sleep on the boat, he found that they were gone to England. A gust of wind from the southwest was pushing them back to where they had originated from. On December 27, after Christmas, the ship finally left. Their very first stop was intended to be Tenerife in the Canary Islands, a similar place that Darwin had always wanted to visit with Henslow. Shockingly, they must be isolated for twelve days before landing, because of the current cholera outbreak in Britain, so the captain gave the request to set sail for St. Jago in the Cape Verde Islands, 300 miles off the African drift. As the ship is moving, Darwin started his work as a naturalist by gathering tiny fish, plankton. When they arrived at St. Jago, he climbed through the well of lava slopes, experiencing his first tropical wilderness in a little valley and seeing genuine proof of geographical change: a layer of compacted ocean shells in the precipices thirty feet above ocean level. On February 8, 1831, they visited at St. Pauls Stones to execute birds for sustenance. After that, they traveled to Bahia. The ship crew spent several weeks in South America before they take off to Rio. His most energizing find was a fossil Megatherium, an extinct ground-abiding relative of the sloth. This was back on September 22. This must be the biggest moment for him because it is rare to found fossils and my favorite part of the book. It is fascinating. Darwin would collect data and samples to send it off to Henslow, his friend. At the beginning of 1833, a year and couple months later, they almost got their ship sunk from an awful climate. They made it securely to the home region of the Fuegians. They had expedited board from Britain, two gentlemen and a lady who had been kidnapped by the captain on a past excursion. They dropped off the Fuegians with an English minister who would have liked to spread Christianity. However, when the Beagle returned nine days after the fact that preachers things had been stolen. The Captain had purchased the second boat in the Falklands. Darwin took off, voyaging 200 miles in two weeks and executing eighty various types of birds and different species. He kept on sending his samples to Henslow. This is the part where it improves my understandings of biology. He used these birds to study different types of finches beaks and how they adapt food based on their environment. The Beagle got to travel south again in the last month of the same following year, passing strange countries on its way through the Straits of Magellan to where the prisoner Fuegians had been dropped off a long time previously. Darwin headed inland towards the Andes with a group of people and tools, yet arrangements ran low and they were compelled to turn back before contacting them. Luckily, he knew he would have an opportunity to contact them from the opposite side when the Beagle went to Chile in South America. On June 6, 1834, they made it around the Horn and touched base at the island of Chiloe, off the west shoreline of southern South America. From that point, they went to Valparaiso on the last week of July. Since it was winter, it was excessively unsafe, making it impossible to achieve the Andes legitimate, however, Charles Darwin made it to the lower regions in August, returning through Santiago. There was a little scary moment for a second: Fitzroy had clearly had a breakdown because of questions about the exactness of his estimations on the eastern shoreline of South America. Luckily, the officers persuaded him to continue his post and it was settled that there was no compelling reason to come back toward the east drift for promoting estimations. Beginning in the spring, he finally accomplished his fantasy of seeing the Andes very close. After coming back from the successful Andes campaign, Darwin rejoined the Beagle for the excursion north to Lima, where they touched base on mid-July 1835. Two months later, they traveled west into the Pacific Ocean to their first look at the Galapagos Islands, which Charles Darwin w as later to make well known, on September 15. Galapagos Islands are an archipelago of volcanic islands in the Pacific Ocean near South America. Charles saw different species of marine iguanas, birds, lizards, and even big tortoises that live on land. The sad part is that they killed at least a dozen of tortoises for samples and specimen. When he moves to different small islands, he noticed that the birds are somewhat different than others. He is very curious why these birds are different than others. He drew different types of finches beaks in his journal. Charles also noticed that the giant tortoise has different shapes of the shell. The left the island on October 20, 1835. In conclusion, The Voyage of the Beagle is not just a vital book in the historical backdrop of present-day thought yet, in addition, an exceedingly critical one in the life of Darwin. As a young fellow, Charles had a little feeling of business or heading. When he was only sixteen-years-old, he started a profession of medicine at Edinburgh College. Finding, in any case, that he was unfit for the calling, he entered Christ School, Cambridge, after three years in 1828 to set himself up to be a pastor. Neglecting to take respects or to separate himself in any capacity, he acknowledged the offer of Captain Fitzroy of the Beagle to sign on as a naturalist on a voyage the world over that, in the end, took five years. Amid that time, Darwin found himself and his profession as well as started mentioning those objective facts that he later formed into the hypothesis of advancement clarified in On the Origin of Species. This work, together with crafted by Karl Marx and of Sigmund Freud, constituted an intense impact on twentieth-century logical idea and qualities. In this book, we have this man whose thoughts have reformed totally our comprehension of life, composing with fascinating about the voyage which started and formed his reasoning regarding the matter. He is one of the historys most influential thinkers.
Sunday, March 8, 2020
The Causal Factors of Divorce in America essays It is hypothesized that one causal factor of divorce among heterosexual American couples is financial problems. It is said that the number one cause of divorce is financial problems. No mater how rich or poor a person is, one constant subject of marital disagreements leading to divorce is money. In reality, of course, it is not the lack of money that causes divorce. The vast majority of couples who split cite money as the main or primary factor. A recent study found that 57 percent of divorced couples said financial disputes were a primary cause of marital discord (Larson, Newell, Topham, Nichols, 2002). Ten percent said it was the main cause of divorce. However, the issue of money doesn't always have to lead to the demise of a relationship. For married couples the financial aspect of the relationship actually has some benefits(Johnson, Wright, Ketring, 2002). There is a sense of stability and security when two people can work together and manage money cooperatively (Johnson, Wright, Ketring, 2002). Married couples also enjoy more tax benefits than singles because they are more likely to own a home than to rent. However, money can cause some serious pr oblems in a relationship. While different views about money can lead to strife, experts say many times the issue of money is only the lip of the iceberg for a mound of problems that exist in the relationship (Johnson, Wright, Ketring, 2002). One money issue for couples of today have to deals with is the women bringing home the larger paycheck (Crouter, Sayer, McHale, 2001). Now more women are working and many of them are holding down high-paying, professional jobs. Women generally are the money managers in the relationship responsible for budgeting, balancing the checkbook, etc. with little or no input from their partner(Lavee & Katz, 2002) . This was not much of a problem for couples say, a generation ago, when the man was the primary breadwinner of the household. The resentme...
Friday, February 21, 2020
Topic Selection and Rationale (Attempt 1 ) - Essay Example The ability to properly define and/or guarantee protections for unique works or concepts could easily lead to stalemate if the intellectual property is governed domestically yet regulated abroad when doing business in foreign nations. In microeconomics, comparative advantage could explain a potential stalemate when it comes to intellectual property protection. This is when a firm has the ability to produce a work or product at a lower opportunity cost than competition. An opportunity cost is when the highest valued alternative must be sacrificed to select another strategy. If one business operates in a market where there are many competitors, and are able to experience lower cost in the action, competition may be more adamant about protecting their intellectual property as a competitive tool. Because the WTO acts as an agency and forum for these discussions, it is likely that the WTO will become engaged in trying to settle the dispute. A competing company might have very high cost objectives, however they cannot lower their opportunity costs to seek a special project or product innovation. Animosities between the rival companies could cause conflict with the membership of the WTO, based on the high cost of doing business against a competing product able to avoid high opportunity costs. In macroeconomics, inflation could also lead to problems with intellectual property rights disagreements that will ultimately involve the WTO. This is when the price of goods and services rises due to supply, finance, or money supply. Companies that are unable to compete effectively against another company that enjoys comparative advantage may also have inflationary issues arising from the cost of doing business which, in turn, affects overhead costs and production costs. The company could, at the same time, be impacted by inflation associated with international distribution.
Wednesday, February 5, 2020
Tacit Knowledge for the Training of pre-service Industrial Design and Technology Teachers in Australia - Dissertation Example From the research it can be comprehended that a lot has been heard and talked about tacit knowledge in the field of teaching and teacher training. According to Chandra and Chalmers, the knowledge contains certain tacit aspects that cannot be codified but are only transmitted through personal experience or training. Tacit knowledge is defined by Shulman as the knowledge that cannot be transferred to other using words either written or verbalised. As such, tacit knowledge has been recognised in the literature of teacher training only one aspect Ã¢â¬â who possesses this knowledge Ã¢â¬â rather than focusing on other aspects and taking it a step further on how this knowledge can be passed down or diffused within the pre-service training period. Various educational theories have cited tacit knowledge as the sole means of learning to teach but this has been a topic of debate and controversy throughout the history. Ma debating on one of the educational theories stated that the traditio nal epistemology is about teaching which include the concepts of objectivist, behaviourist, and positivist. Even though, as stated by Ball, that each of these concepts visualise a different teaching method, the concepts centre on the idea of delivering the content from a tacit point-of-view. In other words, the traditional teaching epistemology focuses on articulating the tacit knowledge rather than to prompt the pre-service teachers to respond or act in a particular manner. (Ball, 2000, p.244). Over the past few years, attention to research on teacher training and education has shifted away from behaviours or skills teaching to the knowledge and beliefs of the teachers i.e. the tacit knowledge (Lampert, 2001, p.127). This shift occurred because of dissatisfied teachers that deliver the teaching contents in a mechanistic manner. In order to understand this behaviour, investigations were carried out in classroom settings that focused around the knowledge and beliefs of the teachers ( Chick, 2002, p.180). Recently, the interest in the training of pre-service teachers has increased because of the concept and utilisation of knowledge. Although researchers take different approaches in their researches and purposes, knowledge is broadly defined as tacit knowledge that teachers learn with respect to their teaching experiences (Sullivan and McDonough, 2002, p.250). Tacit knowledge, for many researchers, implies the presence of complex and context specific teaching nature that can contribute to the enhancement of teachers teaching status as a profession (Chick, 2002, p.187). This tacit knowledge has been explained by many researchers, but was introduced and elaborated by Shulman using the concept of pedagogical content knowledge (Chandra and Chalmers, 2008, p.25). The concept explains tacit knowledge as the interpretations and transformations of subject matter in context that can facilitate student learning (Chandra and Chalmers, 2008, p.26). Teachers who are preparing to teach in secondary schools that often struggle in subjects such as science particularly industrial design and technology. They need to take sufficient courses to meet the degree requirements and learn enough about the subject to be able to teach in schools. However, the training of pre-service industrial design and technology teachers often falls short of preparing the new teachers with the necessary skills and knowledge (Shulman, 1986, p.11). As argued by Ma (1999, p.26), the curriculum changes in the schools particularly in the industrial design and technology subject area, teachers are not being adequately training. The reform in the curriculum requires that teachers need to be trained to be able to provide quality education to all students (Ball, 2000, p.243). However, the exact contents of the subject have been subjected to controversy as when, what, where and how can it be acquired by the teachers. One research suggests that in order to underst
Tuesday, January 28, 2020
Expression of Recombinant Green Fluorescent Protein (rGFP) Expression and Purification of recombinant Green Fluorescent Protein (rGFP) from E. coli using Ni2+-Agarose Column Chromatography. Andrea Bustamante Janakikeerthika Darmarpandi Abstract Green Fluorescent Proteins are vital components of bioluminescence in marine animals. There unique ability to withstand and recover from harsh conditions and regain fluorescence was of great interest. The purpose of the following set of experiments was to express and purify a His6-Xpress epitope tagged recombinant form of Green Fluorescent Protein grown and harvested from E. coli. The desired protein is initially released into solution using the properties of freeze-quick thaw cycles that then help release the contents of the nucleus of neighboring bacteria following a chain reaction. It is then submitted through a Ni2+-agarose affinity chromatography column where the target protein was purified. The resulting wash and elution fractions where run through a Bradford assay, SDS-PAGE/Coomassie blue staining, and a Western blot to determine the molecular weight of the protein to be 32kDa. The overall specific activity was determined to be 433000 RFU/ mg of total protein with a resulting 20 percent purity. The results show that expression and purification of rGFP from bacterial cells was possible. Introduction Aequorea victoria is a jellyfish capable of producing a green fluorescent light when Ca2+ ions activate a photoprotein, known as aequorin, which excites Green Fluorescent Protein (GFP). Wild type GFP is a 27kDa, homodimer composed of 238 amino acid residues that absorbs light at an excitation wavelength of 395nm (blue light) and emits light at an emission wavelength of 510nm (green light). Aequorea victoria GFP has a distinctive three dimensional structure that encases a chromophore (formed by cyclization of Ser65-dehydrogenized Tyr-Gly67) and allows for stability under harsh conditions (Prasher, 229-230.) . This structure allows for regaining of fluorescence even after the protein has been denatured upon removal of the denaturant. Therefore, GFPÃ¢â¬â¢s are extremely stable to changes in pH, temperature, oxidation and reduction, and chemical reagents (Pan, Pickett, and Rippel 225.) Poly-histidine tags involve addition of a series of histidine residues to the N or C terminus of a protein of interest. Poly-histidine tags are affinity tags that serve to facilitate protein purification by exploiting the positively charged histidine residueÃ¢â¬â¢s affinity for negatively charged columns. This series of experiments involved a six repeat histidine codon contained within a DNA plasmid which resulted in a recombinant Green Fluorescent Protein that contained a six residue histidine tag located at the N-terminus. The HisÃ 6 tagged recombinant Green Fluorescent Protein was then subjected to Ni2+-agarose column affinity chromatography. Ni2+-agarose affinity chromatography allows for the purification of poly-histidine tagged proteins due to the selectivity and affinity of the Ni2+-agarose matrix for His6 tagged proteins. rGFP binds the column due to the interactions between the His6 tagged proteins in the mobile phase with the metal Ni2+ ions immobilized within the matrix in the stationary phase. The Ni2+ ions contained within the matrix are capable of binding electron rich molecules including histidine residues and allowing most other molecules to pass unbound. This results in the binding of the desired protein to the column and the purging of most undesired proteins and contaminants from the column into wash fractions (Ninfa, et al. 100-101.) The column was then subjected to imidazole, which competes with rGFP for Ni2+ ion attachment, and this allows for the elution of the target protein. Due to its unique properties, isolation of GFP was of great interest and expression and purification were the main focus of the following series of experiments. A suitable way to accomplish this was devised using the combination of poly-histidine tagging and affinity chromatography. The purpose of this experiment was to express and purify a six-Histidine tagged recombinant form of Green Fluorescent Protein from E. coli through the use of Ni2+-agarose affinity chromatography. After expression and purification, a Bradford assay was performed to estimate total protein amount. This was followed by SDS-PAGE/Coomassie blue staining to determine purity and molecular weight. The confirmation of the presence of rGFP was done using the Western Blot. Materials and Methods Growth of G strain In a test tube, 10ml of liquid LB growth media containing 100ug/ml Amp and 25ug/ml Cam was inoculated with a single bacterial colony of strain G (BL21(DE3)uv>) and was allowed to grow overnight at 37Ã °C. The culture was shaken until saturated. In a flask, 500ml of liquid LB media (pre-warmed to 30Ã °C) was inoculated with about 4 ml of the saturated overnight culture (or until the 500ml culture reached an OD600 reading of 0.1) and allowed to grow at 37Ã °C until the OD600 reading reached 0.5. At approximately OD600 ~0.5, or time zero, 1ml of the culture was harvested into a 1.5ml centrifuge tube and pelleted. The supernatant was discarded and the Ã¢â¬Å"G0Ã¢â¬ pellet stored at -20Ã °C for later use. The remaining culture was induced with 1mM IPTG and allowed to grow. After 3 hours, 1ml of the culture was harvested into a 1.5ml centrifuge tube and pelleted. The supernatant was discarded and the Ã¢â¬Å"G3Ã¢â¬ pellet stored at -20Ã °C for later use. An additional 15ml of the IPTG induced culture was harvested into a 15ml centrifuge tube and pelleted. The supernatant was discarded and the Ã¢â¬Å"G3-15mlÃ¢â¬ was stored at -20Ã °C. Preparation of rGFP Crude Extract Immediately after removal of the Ã¢â¬Å"G3-15mlÃ¢â¬ pellet from freezer, breaking buffer [10mM Tris, pH 8.0; 150mM NaCl] was added into the centrifuge tube. The breaking buffer was pipetted up and down (being careful not to introduce air) until pellet had thawed and homogeneity was reached. The solution was transferred into a 1.5ml centrifuge tube, vortexed for 5 minutes, labeled and placed in 37Ã °C water bath for 10minutes after which the centrifuge tube was transferred to a rotating platform shaker in a dry air 37Ã °C incubator for 20 minutes. After lysis, the mixture was centrifuged at 14000xg, 4Ã °C, for 10 minutes. In a dark room in the presence of a hand held UV light, the fluorescence of the pellet and supernatant where observed the recorded. The supernatant was then decanted and care was taken not to get the pellet back into the supernatant as centrifugation would be required if this did occur. This supernatant was the GCE (rGFP crude extract) Preparation of Ni2+-agarose Column In a 3ml plastic syringe, enough glass wool was placed into the well to cover up to the 1/4 ml marking. The syringe was secured onto a ring stand and placed perpendicular to the ground. About 100ul of breaking buffer was pipetted into the top of a closed luer-lock and allowed to overflow. 1ml of buffer was then pipetted into the syringe column and the luer-lock was immediately screwed onto the syringe. An additional 2ml of breaking buffer was added to the column and several drops of buffer were allowed to flow out. The luer-lock was then returned to the closed position. A total of 500ul of breaking buffer was added to the column and then 1ml of a 0.5ml bed volume Ni2+-agarose slurry was added to the column. The luer-lock was opened and agarose matrix was allowed to Ã¢â¬Å"gravity pack.Ã¢â¬ The column was pre-equilibrated with 5ml of breaking buffer and then the luer-lock was returned to the closed position. Ni2+-NTA Chromatography Separation Procedures 100ul of GCE was transferred into a centrifuge tube, labeled, and set aside. Breaking buffer was added to remaining GCE if content was less than 1ml. GCE was slowly applied to the Ni2+-agarose column and allowed about 5-10 minutes for protein to bind to column. The luer-lock was opened and 0.5ml of effluent was collected into 1.5ml centrifuge tube and labeled W1. This was repeated with the subsequent effluent labeled W2.The column was then observed under an ultraviolet light and fluorescence recorded. The column was then washed with 4ml of buffer in 0.5ml increments. The effluent was collected and labeled W3 to W10. The column was then washed again with a total of 5ml of breaking buffer. This effluent was discarded. A total of 5ml of elution buffer containing 10mM Tris, pH 8.0; 150mM NaCl, 300mM imidazole was added to the column in 0.5ml increments. The eluents were collected and labeled E1-E10.The column was then observed under a UV light and the fluorescence recorded. The W1-W6 and E1-E6 fractions were also observed under UV light and their fluorescence rec orded qualitatively. Determining Total Protein Amount A standard curve was created using six different samples of Bovine Serum Albumin (1mg/ml) of known amount. The amounts of BSA used all had a final volume of 50ul and included 0ug, 3ug, 5ug, 10ug, and 20ug total proteins. A total of 1ml of Bradford reagent was added to each, vortexed, and allowed to incubate for 10 minutes. The results where read using 200ul in a microtiter dish and read using a microplate reader set to 595nm. The results where plotted on a graph as absorbance (595nm) vs. BSA (ug) and a best fit line was drawn. The Bradford assay was then performed once on the W1-W6 and E1-E6 samples. Any samples whose absorbance fell outside the standard curve were repeated less sample in the assay. Once all samples fell within the standard curve, the Bradford assay was repeated two more times for each sample. The total protein amount was then extrapolated from the standard curve using the absorbance values. Estimating Purity and Molecular Weight The SDS-PAGE was prepared using a 12 percent resolving gel that was poured between the Bio-Rad glass plate Ã¢â¬Å"sandwichÃ¢â¬ and allowed to polymerize. A 5 percent stacking gel was prepared and added on top of the resolving gel, a comb was inserted, and the gel was allowed to polymerize. Once that polymerized, the combs were removed and the electrophoresis tank was set up. 15ul of G0, G3, GCE, W3, W4, E2, and E3 samples were added to the SDS-PAGE along with a standard molecular weight ladder. The samples were electrophoresed at 200volts for 45 minutes. The gel was then stained using Coomassie blue dye and the stain removed. Confirmation of rGFP 2-Ã ²-mercaptoethanol was added to the centrifuge tubes containing the G0, G3, GCE, W3, W4, E2, and E3 samples and were loaded along with a molecular weight ladder and electrophoresed as described above. The stacker was removed and the resulting gel set up for transfer onto a nitrocellulose membrane for Western Blot analysis. The overall setup required a Ã¢â¬Å"building upÃ¢â¬ of components with the positive electrode base on the bottom, followed by filter paper soaked in transfer buffer, nitrocellulose paper above that, the SDS/PAGE layer, another layer of filter paper soaked in transfer buffer, Western blot solution was poured over all the components, and finally the negative electrode lid was locked into position. To ensure transfer, the nitrocellulose gel was stained using Ponceau S and allowed to incubate for two minutes on a rocker and then destained using ddH2O. The membrane was then blocked using 5% non-fat dry milk/TBS solution and incubated for 30 minutes on a rocking p latform. This was then and washed three times with 0.05%Tween 20/TBS with 5 minutes of incubation between each wash. It was then probed with mouse IgG anti-Xpress epitope MAb solution and allowed to incubate for 45 minutes. The 0.05%Tween 20/TBS wash was repeated in triplicate. A secondary probe using sheep IgG anti-mouse IgG conjugated horseradish peroxidase polyclonal anti-serum solution was performed as above and then washed in triplicate. The nitrocellulose gel was developed using TMB until desired intensity was reached and development was stopped with water and results recorded immediately. Results The expression of the target protein was doubly repressed in the G0 (uninduced) sample of E. coli. First, the Lac repressor protein binds to the lac operator and prevents transcription by T7 RNA polymerase (Garrett and Grisham 915-916). Second, T7 RNA was repressed by lysozyme protein that binds to T7 RNA polymerase and inhibits transcription. Expression of rGFP in the G3 (3 hour post induction) sample was made possible through the use of IPTG (Garrett and Grisham 914.) The purpose of IPTG was to repress the Lac repressor which resulted in T7 RNA polymerase being able to transcribe DNA downstream of the T7 promoter and expression of His6-Xpress-GFPuv, resulting in the fluorescent capable recombinant Green Fluorescent Protein. (Figure 1) This resulting recombinant GFP is a 279 amino acid protein. rGFP has a six Histidine tag at its N terminus between amino acids 5 and 10, an Xpress epitope between amino acids 24 and 31, Green Fluorescent Protein between amino acids 39 and 277, and a 3 amino acid end tag between amino acids 277 and 279. The chromophore is found between amino acids 103 and 105 in the DNA sequence. (Figure 2) Results of Ni2+-agarose affinity chromatography and Bradford assay indicated that the E3 (elution 3) sample contained the most rGFP activity with approximately 18,600 RFU (relative fluorescent units) and an estimate 43ug of total protein. The specific activity calculated for the sample was 433000 RFU/ mg of total protein. (Figure 3) The SDS-PAGE/Coomassie staining gave an estimate molecular weight for rGFP of 32kDa based on a total traveled distance of 2.3cm along the SDS/PAGE. The overall purity of the band was approximately 20 percent. The higher molecular weight band was most likely contaminants at about 45kDa and the lower molecular weight band was possibly a result of the degradation of the c-terminus at 27kDa. (Figure 4) Western Blot indicated prominent bands in the E3, E2, GCE, and G3 lanes. Lanes W4 and W3 showed very light bands and lane G0 shows an absence of bands. All visible bands appear at about 32 kDa and therefore confirm the presence of rGFP. (Figure 5) Conclusion The successful expression and purification of recombinant Green Fluorescent Protein is significant in the scientific community due to the possible uses for it in the future. Green Fluorescent Protein is significant because it provides an inexpensive and relatively easy method of detection. The possibility for real time detection means result could be obtained in real time. Future experiments will focus on linking rGFP to proteins during transcription and translation. This would result in a desired protein with a GFP tag whose fluorescence can then be used for identification. This should result in the ability to locate a target protein using the fluorescence of rGFP. Future applications of GFP could include incorporation into the genetic code of small mammals. These could encode fluorescent neurons which in turn could help further research in areas such as nerve tissue regeneration or other advances in neurobiology. Its unique properties of endurance could be exploited to understand how it can endure harsh environments and still regain functionality after remediation. This would have significant applications in molecular and cellular biology in understanding cellular degeneration and how help patients with diseases involving cellular degeneration. Bibliography Pan, Jing, Elizabeth Pickett, and Scott Rippel. Biochemistry Laboratory Lecture Notes. Dallas: UTD copy center, 2013. 225-289. Print. Pan, Jing, Elizabeth Pickett, and Scott Rippel. Biochemistry Laboratory Manual. Dallas: UTD copy center, 2013. 38-77. Print. Prasher, Douglas C., Virginia K. Eckenrode, et al. Primary Structure of the Aequorea victoria green-fluorescent protein. Gene. 111. (1992): 229-233. Print. Garrett, R., and Charles M. Grisham. Biochemistry. 4th ed. Belmont, CA: Brooks/Cole, Cengage Learning, 2010. Print. Ninfa, Alexander J., and David P. Ballou. Fundamental laboratory approaches for biochemistry and biotechnology. Bethesda, Md.: Fitzgerald Science Press, 1998. 89-107. Print.
Monday, January 20, 2020
Autism Spectrum Disorder (ASD) is not one specific disorder, but a group of disorders that have similar behavioral characteristic, such as difficulty with communication and socialization. The cause of ASD is still unknown but there are studies that indicate there may a genetic or environmental component. More specifically, ASD can cause limitations in intellectual abilities as well as difficulties with attention and motor coordination. ASD is usually identified during the first three years of life and affects boys at a higher rate than girls (Autism Society). ASD actually occurs at five times a higher rate in boys than in girls, affecting as many as 1 in 54 boys as compared to 1 in 252 girls (Center for Disease Control and Prevention). Newer reports have shown that the prevalence of ASD has increased over previous years and indicates that ASD can actually affect as many as 1 in 88 individuals (Autism Speaks). There are a multitude of different types of therapies than can assist individuals whose lives are affected by ASD. This paper describes many of the challenges faced by individuals with ASD and also explores some of the physical based activities that have been shown to be effective in helping children with ASD overcome many of the obstacles they face. Motor impairments are one of a large number of concerns for children who are diagnosed with ASD. These motor impairments can be quite diverse and include impairments that affect the individual with ASDÃ¢â¬â¢s gross and fine motor skills and control of posture (Bhat, Landa, & Galloway, 2011). There is research reflecting that eighty-three percent of children with ASD demonstrate below average functioning on various motor-skill related tests. These motor impairments can affec... ...dquatictherapy.htm O'Connor, J., French, R., & Henderson, H. (2000). Use of Physical Activity to Improve Behavior of Children With Autism -- Two for One Benefits. Palaestra , 22-29. Prupas, A., Harvey, W. J., & Benjamen, J. (2006). Early Intervention Aquatics. Journal of Physical Education, Recreation & Dance , 46-51. Reid, G., O'Connor, J., & Lloyd, M. (2003). The Autism Spectrum Disorders: Physical Activity Instruction Part III. Palestra , 20-48. Smith, B. (2011, April). Hippotherapy: Teaching Strategies for Students with Autism. Retrieved March 23, 2012, from Bright Hub: http://www.brighthub.com/education/special/articles/19121.aspx Tomchek, S. D., & Dunn, W. (2007). Sensory Processing in Children With and Without Autism: A Comparative Studing Using the Short Sensory Profile. The American Journal of Occupational Therapy , 190-200.
Saturday, January 11, 2020
Introduction The term Ã¢â¬Å"disasterÃ¢â¬ has only transient significance. It is a set off, a flag to signal a meaning, a spur to produce a specific reaction. Yet it has almost no significance for the activities that are carried out under the marque of a disastrous event. Disasters do not cause effects. The effects are what we call a disaster (Dombrowsky 1998, 21). No disaster Ã¢â¬Å"worksÃ¢â¬ and an earthquake is not anything more than shock waves, never a fist that hits a city. The term that Ã¢â¬Å"a disaster strikesÃ¢â¬ is as wrong as saying Ã¢â¬Å"the winds blow,Ã¢â¬ as there is no separate process that swells the cheeks to rage. Wind is air in explicit motion, not a separate being that makes the air move. In a definite way, it is the same with disasters: there is no difference between a disaster and its effects. And to cope with disaster, Disaster Management came into existence. Crozier and Friedberg characterize a Disaster Management organization as a Ã¢â¬Å"merger of human beings with the aim to solve their vital problemsÃ¢â¬ (1979:12). The capabilities of the organization for problem-solving, however, develop step by step from those solutions that have been successful in the first place. Read more:Ã Sharing Responsibility During Disaster Disaster Management, as Crozier and Friedberg put it, and then lean to organize themselves around their success. In the long run, the successful solutions particularly have to be defended against competitors and envy. Thus, the operation of organizations leans to turn into a hedgehog position. Progressively, the operation of the organization is transferred into the centre of the efforts of its members. at that time, an organization has turned to selfishness. Its only interests are self-preservation; the organization is controlling itself, with the original reason for its establishment being turned into a subordinate, accessory matter. Simultaneously, the discernment of reality changes from a creative, problem-oriented awareness, toward a cynical, solution-oriented persistence. Instead of scanning for upcoming problems, the self-preserving organization describes reality within the framework of its available resolutions. The interest is less in focusing on potential solutions for upcoming problems, but more on the applicability of the available solutions. This shift is significant because it marks a difference in the ways of perceiving the world. The primary way is analyzing the problem in favour of finding a passable solution. Thus, basic aim of interagency disaster management is not to focus on the fundamental problems of the victims, but on the solutions they have at hand or can offer. Reality then is completely seen from one approach; the solution defines the problem, and deductively, reality. Ã¢â¬Å"Disaster management Ã¢â¬ are mainly defined this way. The cases where warm clothing was sent to African famines or thousands of tons of contraceptives or cough mixtures were sent to mass victim situations are not only mistakes, but the rational outcome of the inner dynamics of self-preserving organizations. The concept is characteristically used in disaster medicine and in emergency planning for nuclear accidents. Also directly influenced by practice are those conceptualizations of disaster that are built along the distinctive planning and action systems of emergency management. Successful community-based and participatory approaches to disaster management often distinguish local human capacity as well as susceptibility. Empowering those most pretentious by disaster through a role in management, planning and response can have psychosocial and practical gains. People exposed to disaster might have come close to death, which might shake their belief in themselves and their principles to the foundations. Life might be perceived as chaos, diminishing their ability to respond adequately to the new situation, manage their lives and meet basic needs. Rebuilding the foundation will form meaning from chaos, and much of the foundation in life is build on human relations.