Analysis of Proteins in Fish Muscle Tissue

Analysis of Proteins in Fish Muscle Tissue Introduction In vertebrates, the muscular system is an anatomical organ system controlled through the nervous system. Derived from the mesodermal layer of embryonic germ cells, these contractile tissues-of skeletal, smooth, or cardiac origin-are responsible for blood circulation, internal organ function, heat production, and organ protection.[1] With the skeletal system integrated, voluntary and reflexive movement, as well as posture and body position, become possible. Surrounded by an epimysium, skeletal muscles are composed of many long muscle fibers lined with endomysium, which are bound together by perimysium into bundles called fascicles.[2] Within these myocytes, there are smaller strands of myofibrils that contain myofilaments (or sarcomeres) the basic unit of a striated muscle tissue. These repeating sarcomeres contract in response to nerve signals by means of sliding filaments: actin and myosin. The thin filaments consist of two chains of spherical actin proteins twisted in a helical co nformation and troponin as a contraction regulator.[2] Each actin molecule has a myosin-binding site that is covered by tropomyosin during muscle relaxation. Having a head and tail region, myosin II proteins generally form the thick filaments with its six polypeptide chains and can cross bridge with actin filaments due to their elasticity and contractibility properties. Specifically, the motor domain of its two heavy chains adopt an ÃŽÂ ±-helical coiled coil configuration and couple ATP hydrolysis with its motion while its two light chains-which wrap around the neck region of each heavy chain at the IQ sequence motif-have regulatory roles[1]. Although this major multi-subunit protein has remained greatly stabile across the animal kingdom over time, myosin light chains have undergone evolutionary divergences for different species; however, the essential structure and functions have remained highly conserved.[3] Caused by genetic mutations, only favorable variations are passed thro ugh this process allows for specialization, speciation, and evolution that eventually increases survival ability: DNA (genes)  ® RNA ® Protein  ® Trait  ® Evolution. Protein gel electrophoresis and western blotting can be used to compare myosin light chains of different species by identifying any commonalities or alterations in specific subunits. Since proteins reflect changes in the gene pool, the phenotype and function as well as form of an organism can be identified, allowing for the study of their physiological adaptations to the environment. Through comparative proteomics-defined as the analysis of differentially expressed proteins with comparison between at least two protein profiles-changes in the proteome that have been caused by development, diseases, and the environment can be identified allowing for assessment of biological variability and dataset comparability.[4] The objective of this lab was to extract proteins from unknown samples of fish muscle tissue and then qualitatively analyze this protein mixture by performing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) twice. The protein bands of the first gel-representing the total amount of proteins found in the tissue homogenate-were stained and visualized at 595nm with the Bio-Safe Coomassie Blue G-250 dye at 595nm while the fractionated proteins of the second gel were electroblotted onto a nitrocellulose membrane via Western blotting where the specific protein of interest was selectively immuno-detected by chemiluminescence with a horseshoe radish peroxidase-linked secondary antibody. [3,4] Accordingly, the goal of this report is to identify the different types of proteins found in fish muscle-specifically of shark, tilapia, skitter, and salmon-required for muscle contraction and movement and to establish whether they are highly conserved or variable across all animal species. Consequently, information about the environment, niche, or physiological stresses faced by the organism can be elucidated as specific protein modifications that alter muscle function and performance work to increase their fitness and adaptiveness.[2] Differences in proteins may reveal information about the evolutionary relationships among various organisms and by understanding this diversity in the natural world, many biological problems can be solved to improve the quality of human life.       Materials and Methods First, unknown tissue samples from two different fish species were prepared for protein extraction: in a 1.5mL microcentrifuge tube, 250ÃŽÂ ¼L of Laemmli (1x SDS) sample buffer was added as well as the minced tissue. After gently agitating the contents by flicking the tube, it was left to incubate at room temperature for five minutes. Next, the tube was centrifuged to pellet the tissue; this allowed for transfer of the supernatant buffer to a new 1.5mL screw cap tube, which was then boiled at 95 °C for five minutes. Second, SDS PAGE was performed on two separate precast TGX gels (purchased from Bio-Rad) since both Coomassie Blue staining and Western blotting were required. Refer to the BIO314 experiment 7 lab manual for instructions on how the gel apparatus was assembled with the Mini-Protean gels and tetra cell. When this was completed, the loading scheme for Coomassie staining involved pipetting the protein ladder (Biorad cat #161-0375) in lane 1 (at 7 ÃŽÂ ¼L/line) and the actin/myosin standards in lane 6 (at 5 ÃŽÂ ¼L/line). The rest of the lanes were used to load the samples (at 10ÃŽÂ ¼L/line). The same set-up was done for the immunoblotting gel, except only 5ÃŽÂ ¼L/line of each boiled sample was loaded. Refer to the BIO314 experiment 7 lab manual for instructions on how these solutions were loaded. After all of the samples have been loaded, the gel box lid was connected to the electrode assembly by matching the red and black leads with their corresponding electrodes. Then, the leads were plugged into the power supply, which was subsequently turned on and set to run at a constant voltage of 200V. This process was terminated at 30 minutes when the loading dye started to exit the gel. Refer to the BIO314 experiment 7 lab manual for instructions on how the gels were removed. Third, Bio-Safe Coomassie staining was done on the appropriate gel-with samples loaded at 10ÃŽÂ ¼L/line-which was peeled from the plate: it was then inserted into a container of deionized water and washed for 5 minutes on a rocking platform. Afterwards, the gel was transferred to another container with Coomassie staining solution again, this was left on a rocking platform for 15 minutes. Upon completion, the stained gel was put in deionized water (destaining solution) and the lid was capped onto this container, which was placed onto the rocking platform for 15 minutes. Fourth, the immunoblot was prepared and transferred: with blunt-ended tweezers, the PVDF membrane and bottom stack was placed on the cassette base; the membrane was left facing up. Any air bubbles seen were immediately removed with a blot roller. Since one mini gel was employed, the stack was centered in the cassette. Then, the second gel-with samples loaded at 5ÃŽÂ ¼L/line-was peeled from the plate (from the SD S-PAGE step) and stacked over-top of the PVDF membrane. Any air bubbles present were subsequently removed using a blot roller. Next, a second wetted top-ion transfer stack was placed above this gel. This assembled sandwich was rolled thoroughly with a blot roller to prevent any air bubbles from being trapped. Finally, the lid was closed and locked onto the cassette and this was set inside the turbo blotter to initiate the transfer. When the electro-transfer process was finished, the blots were dismantled and stored (at -20 °C) according to the instructions written in the BIO314 experiment 7 lab manual. After one week, the Western blot-that had been rocked on a platform with block solution A for 1 hour-was placed into 10mL of blocking solution B and 5ÃŽÂ ¼L of primary antibody was added on that solution with swirling; this was incubated for 20 minutes. Upon completion, the gel was washed with 15mL of wash buffer (three times, each with 10 minutes of incubation); then 15mL of blo cking solution B and 5ÃŽÂ ¼L of secondary antibody was added and incubated at 15 minutes. The three wash steps were repeated. With the wash buffer drained, the membrane was put on a plastic paper protector (with the protein side up) and 400ÃŽÂ ¼L of substrate (made by mixing reagent A and B in 1:1 ratio, 200ÃŽÂ ¼L each) was spread evenly across the middle of the blot. A plastic protector was then added over it and this was imaged with a digital imager for chemiluminescence detection and analyzed using the BioRad ChemiDOC-MP Imaging System for the molecular weight and signal intensity of the protein bands (refer to the instructions posted on blackboard on how this program was operated). Results and Discussion According to the Coomassie-stained gel, the variability in the staining intensity of the protein bands in lanes 2, 3, 4 and 5-for skeletal muscle tissue samples from shark, tilapia, skitter, and salmon-signify the difference in the relative abundance of individual polypeptides in each organism (note that lane 5, band 11 was used as the reference). Influenced by factors such as protein expression and control, these species have generated different quantities of proteins with similar masses in their muscle tissues as they have adapted to specific environmental and biochemical interactions.[5] In figure 1, the potential mass and intensity values of myosin-light chain (MLC) are as follows: shark (15.43kDa at 0.37, 17.65 at 1.71, 20.64 at 1.09, 21.60 at 0.25, 23.05 at 0.69, 23.79 at 0.92, and 25.54 at 1.02); tilapia (15.33kDa at 1.34, 16.42 at 0.75, 19.02 at 0.35, 20.37 at 1.56, 21.47 at 0.34, and 23.79 at 0.36); skitter (15.92kDa at 2.09, 17.99 at 0.94, 20.12 at 0.48, and 23.75 at 0.55) and salmon (16.07kDa at 1.13, 20.12 at 0.31, 21.08 at 0.64, 21.76 at 0.26, and 24.92 at 0.34). Due to selective immunodetection of MLC proteins in Western blotting by a primary antibody, the various protein bands lying in the general MLC range of 15-25kDa in the Coomassie gel can be narrowed to: shark (23.94kDa at 1.33); tilapia (24.47 at 0.70); skitter (24.47 at 0.36); salmon (24.47 at 0.22) and myosin marker (24.47 at 2.40) all of which resemble the myosin light chain isoform I (>20kDa) as isoforms II (20kDa) and III (15kDa) have lower masses; with a greater variability of myosin, tilapia has an additional band of 20.68kDa at 0.39 that resembles isoform II. [5] The other bands were dismissed as non-specific background interferences (note that lane 4, band 5 was used as the reference for the immunoblot). The high specificity of primary antibodies in probing their target allows for its wide-use in proteomic research as a reliable immunodetection technique; since proteins can indica te evolutionary relatedness or the presence of genetic diseases, their role as biomarkers has allowed for measurements of physiological changes as well as their quantifications.[6] In the appendix, all of the protein bands for the four species have been assigned a protein that corresponds to its molecular weight. From this, it can be denoted that sharks are more closely related to salmons than tilapia and skitters, both of which are tied for second place. However, based on fish phylogeny: sharks and skitters-belonging to the same class called Chondrichthyes-have diverged prior to the class of Actinopterygiis, which include both salmon and tilapia.[7] In terms of classification relative to the order, sharks (of Elasmobranchii) have the greatest evolutionary relationship with skitters (of Rajiformes), then salmons (of Salmoniformes), and lastly tilapia (of Perciformes).[7] As a hexameric ATPase cellular motor protein, myosin is composed of four light chains (MLC)-two non-phosphorylatable essential alkali chains, two phosphorylatable regulatory chains-and two heavy chains (MHC). Specifically, the protein bands of these light chains have a molecular weight as a range from 15 to 25kDa; this diversity in the masses occur largely from alternative RNA splicing mechanisms that generate multiple tissue-/developmental stage-specific isoforms.[7] Although these polymorphic variations do not significantly alter the actin-activated ATPase activity of the myosin-heavy chain, they affect the actin-filament sliding velocities and kinetics-leading to different force-generating abilities.[8] In an evolutionary context, the existence of these hybrid molecules has been adopted by muscles-in response to changing functional demands-to shorten this translocation time in order to increase their overall fitness. Consequently, numerous variants of slow and fast light chains we re developed despite the underlying plasticity of striated muscles.[7] Voluntary muscles are divided into slow twitch and fast twitch muscles. The main difference is that the former red muscle contracts for longer periods of time with little force, require an oxygen-rich operating environment, and contain only two distinct light chains while the latter white type contracts quickly and powerfully for only short bursts of anaerobic activity as they become exhausted due to lactic acid buildup, have glycogenolytic capacity, and possess three different light chain subunits.[8] Over 90% of swimming muscles from sharks are composed of myotomes that can create massive propulsive forces by contracting their high numbers of white fibres; only a few such as the Great White incorporate bands of red muscle to elevate endurance over strength.[9] Accordingly, this explains why the MLC band on the Western blot has the greatest intensity of 1.33 relative to the other species. Conversely, fish species are generally composed of endothermic red-segmented muscles in their t runk musculature-allowing for their stiff-bodied, slow undulatory swimming motions.[6] Due to their decreased mass of white muscles, MLC bands of tilapia, skitter, and salmon are of lower intensity at 0.70, 0.36, and 0.22 respectively. Relative to mammals, fish myosins share the same light chain patterns but have higher variability in MLC mass and quantity due to adaptive differences in movement between red and white myofibrils.[6] Since they have larger phylogenetic diversity, there is an enormous range of contraction speeds and swimming styles among homologous muscles.[6] For example, fast twitch muscles of rabbit, sheep, and chicken have three light chain components at 250kDa-whereas only one is found homologous at 180kDa among pike, dogfish, mackerel, angler-fish, and carp.[5] Moreover, their poikilothermic-nature may have contributed to these light chain divergences as they were forced to adjust to fluctuating environment temperatures that required specific muscle responses fo r survival.[9] Sources of errors with the techniques employed contributed in hindering the accuracy of the results. First, the amount of protein stained with Coomassie dye varied greatly between the sample replicates since the dye may complex with the anionic detergent in its free cationic form interfering with protein concentration estimates. Moreover, this dye selectively targets amino acid resides arginine, tryptophan, tyrosine, histidine, and phenylalanine; however, the assay performed responds primarily to arginine residues eight-times higher than other ones listed above.[2] Second, reproducibility of the sample preparation and protein extraction steps was an issue due to variability among the skills of the student, which may have caused the quantity differences seen among the replicates. For example: if more tissues were added for one specie, the increased concentration of proteins loaded into the lane would be misled for a true difference in expression among or between the species. To over come these problems: one, an automated protein extraction systems should be employed since its robotic liquid handing technology can control for errors and contaminations leading to greater reproducibility and accuracy; two, silver staining can be substituted for Coomassie due to its higher sensitivity (0.2ng versus 7ng respectively); third, adjustable single-/multi-channel Rainin electronic pipettes should be used as its fully automated and repetitive micro-pipetting has superior consistency allowing for higher throughput work.[4,5,6,9] Overall, it has been discovered that-irrespective of muscle tissue origin-myosin light chain molecules are heterogeneous in mass and intensity and the existence of phasically active fast muscles versus slow tonic muscles has led to characteristic light chain patterns among different fish species. Based on similarities and divergences in the overall protein content and intensities of the different fish species mentioned above, sharks are deemed to be more closely related to salmons than tilapia and skitters both of which are tied for second place. However, according to fish phylogeny, sharks and skitters have diverged before salmon and tilapia, leading to an order classification of sharks (Chondrichthyes, Elasmobranchii) having the greatest evolutionary relationship with skitters (Chondrichthyes, Rajiformes), then salmons (Actinopterygiis, Salmoniformes), and lastly tilapia (Actinopterygiis, Perciformes). Radical alterations in their muscle proteome may have originated from adaptive responses to environmental stresses-i.e. osmotic, anaerobic, and thermal condition changes- or during symbiosis and development since cells can make different sets of proteins based on its specific spatial-temporal conditions.[5] The inferences made in this lab come with great uncertainty due many accuracy and reproducibility problems. Thus, fluorescence two-dimensional differential gel electrophoresis can be substituted for SDS-PAGE; high-throughput proteomic technologies like micro arrays, mass spectrometry-based methods, protein chips, and reverse-phased protein-microarrays can be used for protein profiling and detection; and hybrid separation-analysis techniques such as reversed-phase chromatography-ESI ionization online analysis systems can be utilized for greater sensitivity, accuracy, and precision all of which allow an experimenter to draw firmer conclusions. References Bandman, E. et al. Developmental Appearance of Myosin Heavy and Light Chain Isoforms in-Vitro and in-Vivo in Chicken Skeletal Muscle. Developmental Biology. 1982, 2, 508-518. Chatfield, S. Experiment 7: Extraction and Electrophoresis of Proteins: Immunoblot Preparation. BIO 314 Laboratory Manual. 2017. Chatfield, S. Experiment 8: Development of Immunoblots (Western Blots). BIO 314 Laboratory Manual. 2017. Focant, B. et al. Subunit Composition of Fish Myofibrils: The Light Chains of Myosin. Journal of Biochemistry. 1976, 110-120. Lowey, S. et al. Function of Skeletal Muscle Myosin Heavy and Light Chain Isoforms by an in Vitro Motility Assay. The Journal of Biological Chemistry.1993, 268, 20414-20418. Lowey, S. et al. Light Chains from Fast and Slow Muscle Myosins. Nature. 1971, 81-85. Syme, D. et al. Red Muscle Function in Stiff-Bodied Swimmers: There and Almost Back Again. Philosophical Transactions of the Royal Society B: Biological Sciences. 2011, 1507-1515. Tomanek, L. et al. Environmental Proteomics: Changes in the Proteome of Marine Organisms in Response to Environmental Stress, Pollutants, Infection, Symbiosis, and Development. Journal of Animal Science. 2003, 373-390. Young, R. et al. Structural Analysis of Myosin Genes Using Recombinant DNA Techniques. Journal of Animal Science. 1968, 259-268.

Gus germs and steel Essay -- English Literature

Gus germs and steel 1. Yali asks Diamond, â€Å"Why is it that you white people developed sp much cargo and brought it to New Guinea, but we black people had little cargo of our own.† 2. Jared Diamond’s thesis seems to be that external factors such as geography can affect the fate of human societies. In other words, what separates the winners from the losers is geography. Chapter 1 1. â€Å"The Great Leap Forward† is when human history developed about 50,000 years ago. 2. The giant moas in New Zealand and the giant lemurs in Madagascar were exterminated by humans. 3. 15,000 years ago the American West looked like Africa’s Serengeti Plains. It was filled with elephants, giraffes, zebras, and other African animals. All of these animals living in the American West were either killed off by Clovis hunters or died due to drought. 4. All of the giant animals residing in Africa were able to survive because there were no extreme weather conditions and there were not nearly as many deadly hunters in Africa as there were in America. Chapter 3 1. Pizzaro’s capture of Atahuallpa â€Å"offers a broad window onto modern history† because it has happened many times since then. 2. Technology, or the guns and the steel, was used in exterminating the Incas. The germs that the Spaniards brought over on their horses produced small pox. 3. Diamond refers to the battle at Cajamarca a collision because two of the greatest empires â€Å"collided† in a huge fight. Chapter 4 1. Societies with successful food production would grow because there was enough food for everyone. The greatest food producers became the world conquerors because they were a big society with big ideas for technology. 2. The development of diseas... ...teel, I would have to say that the most decisive factor would be the germs. A good example is Smallpox; the virus alone killed more people than either guns or steel. 4. The East lost its enormous lead to the West (Europe) because Europe developed a merchant system, capitalism, and patent protection for invertors. Europe also did not have a dictatorship like China, so inventors in Europe were worry free. 5. China lost its technological lead to Europe because they were a dictatorship and had high taxations. 6. China’s connectedness became a disadvantage for them because their fertile crescent had no other geographical advantages other than domesticating wild plants and animals. China also had an absolute despot that was controlling the country. 7. The histories of the Fertile Crescent and China hold important lessons for the modern world including.

Explain and evaluate claims made by linguistic relativists regarding the relationship between language, thought and culture Essay

The relationship between language, culture and thought has been a controversial discussion over decades. Many linguists and thinkers have argued that language lead to large differences in culture and thoughts. Some argue that language controls people’s view and thought of the world, where language embodies worldview, and some argue the otherwise. Language, culture and thought may always refer as together, but any one of them implies the other two. In this essay, I am going to focus on linguistic relativism and I will evaluate claims made by linguistic relativists. Linguistic relativism is a weaker interpretation of linguistic determinism. It is â€Å"a window through which to view the cognitive process, not as an absolute. It’s set forth to be used in looking at a phenomenon differently than one usually would. † (Badhesha, 2001) Linguistic categorize and usage influence thought and certain kinds of non-linguistic behavior. Linguistic relativity hypothesis has always been a controversial and serious topic. In late-eighteenth and early nineteenth century, Boas claimed that there’s no intrinsic relationship between culture and language. (Boas, 1911) Acquainted with Boas, Edward Sapir was impressed with Boas’ statement. Later, he proposed a theory which becomes the most famous attempt in demonstrating relationship between language, culture and thoughts, â€Å"Sapir-Whorf Hypothesis† (Whorfian Hypothesis). The Sapir-Whorf Hypothesis can be broken into two simple concepts: Linguistic Determinism and Linguistic Relativism. Linguistic determinism holds a stronger view. It refers to â€Å"the concept that what is said has only some effect on how concepts are recognized by the mind†, â€Å"A strict view that what is said is directly responsible for what is seen by the mind. † (Badhesha, 2002) This stronger point is supported by Sapir. Sapir: â€Å"language and culture are not intrinsically associated† but â€Å"language and our thought-grooves are inextricably interwoven, [and] are, in a sense, one and the same† (1921: 228, 232) Sapir believes that language and culture are not explicitly related but language, culture and thoughts are interwoven that cannot be unwoven each other. Sapir also expressed his view that language affects how we perceive the world, â€Å"Even comparatively simple acts of perception are very much more at the mercy of the social patterns called words than we might suppose. †¦ We see and hear and otherwise experience very largely as we do because the language habits of our community predispose certain choices of interpretation. † (1929, p. 210) Sapir here explained that language determines our thoughts and culture, it affects our views of the world and culture is a product of language. Sapir’s student, Benjamin Lee Whorf supported and made his theory stronger. Whorf’s claims are both to the extreme, strongest but to the weaker and more cautious at the same time. The extreme perspective is linguistic determinism, where the weaker is linguistic relativism. In fact, linguistic relativism is widely spread through Whorf’s work. Whorf: â€Å"The background linguistic system of each language is not merely a reproducing instrument for voicing ideas but rather is itself the shaper of ideas. † (1940, â€Å"Science and Linguistics,† Technology Review 42: 229-31, 247-8) In this quote, Whorf proposed the stronger form where language determines thought, language shapes our ideas. â€Å"The world is presented in a kaleidoscopic flux of impressions which have to be organized largely by the linguistic systems in our minds. † (Whorf, 1940a:231) This is a weaker form where Whorf argue that the world is somehow under the influence of our linguistic systems. Both in stronger and weaker form, Sapir-Whorf Hypothesis argue that our thoughts, ideas, behavior and culture are under the influence of language. This hypothesis is supported with evidence. Perception of colors is one of the obvious evidence that support language influences our thoughts. Different languages has their way to encode and categorize colors, in English, there are sixteen basic colors and common two color terms â€Å"dark† and â€Å"light†. However, Russian has different words to distinguish light blue (boluboy) and dark blue (siniy). When Russian and English speakers are put to a test to discriminate the two blue colors, Russian is found to be better at it. As Russian perceive the two blue colors as different colors unlike English speakers who categorize it as blue generally. In both English and Chinese, when giving directions verbally, we tend to use â€Å"left† and â€Å"right† instead of E/W/S/N, this is also true in many other languages. However, Kuuk Thaayorra (aboriginal language spoken in Queensland, Australia), they use E/W/S/N to represent â€Å"left† and â€Å"right†, e. g. when they refer to right hand, they may say east hand. Their perception of the world differs from us due to the use of direction terms. To them, the world needs to include precise orientations. This is an example of perception of space which display language affects our perceptions and thoughts. Grammatical feature is another proof of language affects our culture. Whorf asserts that â€Å"users of markedly different grammars are pointed by their grammar towards different types of observations†¦ and hence are not equivalent as observers but must arrive at somewhat different views of the world† (Whorf 1940b:61) Whorf suggested that speakers of different languages will think about the world differently. Hopi language (aboriginal language spoken in Arizona) is early evidence to Whorfian Hypothesis on language and thought. In Hopi language system, they don’t have tenses such as –ed, -ing, -s in SAE [1], they have different perspective of time from SAE speakers. Also, in Hopi, the concept of time cannot be counted and talked like a physical quantity. Language also affects and reflects our culture and values. In Hong Kong, we refer to relatives in different terms, not only â€Å"aunt/uncle†, â€Å"cousins† unlike English. For example, we have different terms for cousins that are older and younger than us, e. g. â€Å"biu-gor† (older male cousin), â€Å"biu-mui† (younger female cousin). It reflects the hierarchy and projects the importance of respecting senior in Chinese family. We can see that Chinese family, including families in Hong Kong divides and identifies its members. For Chinese people, they perceive seniors as people to respect. Sapir-Whorf Hypothesis is the idea that the language that person speaks will affect their perception of the world and accordingly their behavior and culture. In stronger form, language determines the way we think and what we are capable to think of where as linguistic relativity; the weaker form indicates that our thoughts and culture are under the influence of language. Although there are more and more scientific and real life examples that support Sapir-Whorf Hypothesis, anthropologists and linguists still argues weather the hypothesis is too strong or not today. Many argue that instead of language determining our thoughts and culture, they are in fact inter-related and none of them should be dominating the other two. The Sapir-Whorf Hypothesis has received criticism over the years, and it summed up to be three main criticisms, the idea of causality, the methods and translations. While Whorf and Sapir claimed that language affects our perception of the world and the formation of our cultures. Both of them didn’t include the evolution of language, what made up our language, the cultural values that may have been included while setting language system. It is possible that cultural values that might have developed language helped the way we perceive the world. Humboldt (late 18th century) also questioned Sapir-Whorf’s hypothesis, â€Å"The spiritual traits and the structure of the language of a people are so intimately blended that, given either of the two, one should be able to derive the other from it†¦ language is their spirit, and their spirit is their language; it is difficult to imagine any two things more identical. † He also questioned the causality of such hypothesis. Whether if language is the spirit of thoughts and culture or the otherwise. He claims that it is uncertain which one should be derives and to be the spirit of the others. Another obvious criticism is the methods Whorf’s methods. Some believed that Whorf deliberately translated Hopi language in a certain way to support his own hypothesis, to emphasis another thinking system. Linguists, Steven Pinker accused Whorf’s method with strong attitude, â€Å"No one is really sure how Whorf came up with his outlandish claims, but his limited, badly analyzed sample of Hopi speech and his long-term leanings towards mysticism must have helped. † (Pinker, 1994) He also overthrown Whorf’s claim of Hopi people has different perception of time as â€Å"anthropologist Malotki (1983) has found that the Hopi do have a concept of time very similar to ours. † (Neil Parr-Davies, 2001) Translation is another criticism of Sapir-Whorf Hypothesis. Many critics raised the question, if our thoughts are really affected and determined by language, then presumably certain concepts would only be understandable to people that shared the same language. That suggested that if the hypothesis is entirely true, Whorf would have been failed to understand Hopi people’s concept, needless to say even to understand their first thought. Although criticisms are raised against Sapir-Whorf Hypothesis, there are more evidence and claims that support it even in modern days. In fact, psycholinguistic have been studying far beyond perception of color and Hopi language currently, it has been studying and discovering more evidence in modern days such as emotion perception, memory etc. Modern famous researchers like Lera Boroditsky, John A. Lucy believe in Sapir-Whorf Hypothesis, that language indeed influences thoughts, however in relatively narrow ways. I agree the Sapir-Whorf Hypothesis to a large extent. I believe language does influence our thoughts and perceptions of the world. Recently, I have been studying different accents in the world and discovered that it is evidence supporting the linguistic relativists. For example, if someone speaks English in BBC accents or Queen’s English, we would immediately refer them coming from higher social status in Britain. Indeed, our perception can be easily affected by the trivial elements in language as accents. Even nowadays, there are more and more evidence that prove linguistic relativists; we cannot deny the fact that language does somehow shape our daily thoughts and life. Language changes the way we see different culture, it can reflect a certain culture and background of people. Taking Soviet Ukraine as an example, as the prices and supply of product was centrally controlled, they were very cheap and hard to find, and instead of saying â€Å"What are they selling? † they ask, â€Å"Shcho dajut? †(What are they giving? ). Through the language, we can see the cultural values and background of a country, allowing us to understand and perceive different cultures. Nonetheless, I disagree to a small extent as I believe, on one hand, language may shape our thoughts and culture; on the other hand, our culture and thoughts also shape language. Language may act as a tool merely reflecting our thoughts and culture. Under patriarchy and stereotyping, many European countries referred male as a stronger, rational and more dynamic member of the society while female were emotional, silent and subtle one. Leading European languages like French, Spanish are some examples that reflect social stereotyping under patriarchal influence. In French, almost all sport and daylight activities are referred as male, â€Å"le soleil† (the sun) while almost all night time activities are referred as female, â€Å"la lune† (the moon). Sapir-Whorf Hypothesis is a powerful and leading linguistic theory that suggests the relationship between language, culture and thoughts. It can be interpreted as linguistic determinism and relativism; it remains arguable and controversial whether this hypothesis is too strong. Much evidence arises even until today supporting the hypothesis. It is inevitable that every hypothesis receives criticism, but I truly believe the reason why Whorfianism is still being studied today is because we can never deny the influence of language on our thoughts and culture. Language may not be the one the strongly defines our perception and values, but it does affects us in our everyday life.

Report on the Security System at Natividad Medical Center

Security plan This final part of the project contains an in-depth and comprehensive report on the security system at Natividad Medical Center. Using relevant peer-reviewed and technical reports, I devise an analysis plan that explains thoroughly, how I will analyze as well as evaluate Natividad Medical Centers Hospital Computer Information Systems (HCIS) network and its hospital-grade systems infrastructure and technology components. In this comprehensive report, the details of what is going to be analyzed is presented as well as how I am going to analyze the systems (process and tools). The criterion for measuring the various elements is also presented. The rationale behind the choice of the elements is also presented. What is to be analyzed in the system? The analysis is going to involve a scrutiny of the Natividad Medical Centers Hospital Computer Information Systems (HCIS) network and its hospital-grade systems infrastructure and technology components. The analysis is going to be carried out in line with the requirements of Patient Safety and Health Insurance Portability and Accountability (HIPAA) Acts, the Joint Commission (TJC) as well as other national and international regulatory bodies. All elements of information assurance and security are to be ascertained. In this regard, information availability, confidentiality, integrity and security are our primary concerns (Dennis,2005).As indicated earlier, the hospital industry is governed by tough laws andShow MoreRelatedSystem Analysis and Recommendation Report of Natividad Medical Center859 Words   |  3 PagesSystem analysis and recommendation report In this section, I present a system analysis and recommendation report on the Natividad Medical Centers Hospital Computer Information Systems (HCIS) network and its hospital-grade systems infrastructure and technology components. The system analysis report details the findings of the system analysis in the part of system vulnerability/risk assessment as a critical component of the security plan. Why the system vulnerability/risk assessment was carriedRead MoreRisk Assessment Tools1220 Words   |  5 Pageshowever, they perform them differently (Schreider, 2003). Each product is a questionnaire based on the type of organization, asset value, etc. More sophisticated products also allow importation or links to data from penetration tests, intelligence reports, and other risk-gathering formats. They also perform calculations for risk probability and rank each risk by level of importance. The comprehensive design calculates loss estimates, such as ALE, and financial metrics (cost of mitigation and returnRead MoreThesis About Call Center Agents14127 Words   |  57 Pagesreview mates in a local review center headed to many local BPO companies here in our city. BPO or business process outsourcing involves the contracting of the operations and responsibilities of specific business functions (or processes) to a third-party service provider. Employees who work under these companies are referred as call center agents or customer service representatives. Any graduates here in the Philippines has once in a while heard things related to call center companies, etc. Undergraduates