Techniques for Studying Gluten Protein-Free-Samples for Students

Question: Find a protein of interest and describe five Protein Technologies that have been used to study it. Answer: Protein: Gluten Gluten comprises of two different protein components; gliadin and glutenin. It is the composite of both glutelin and prolamin proteins that are conjoined to starch in grass-related cereal endosperms. Gliadin has between 30000Da and 80000 Da of relative molecular mass while glutenin has several millions Da of relative molecular mass (Hochegger et al, 2015).Gluten is thus a long molecule with strong but flexible characteristics. Structurally, gluten is a yellowish-white, elastic, sticky and tough protein. Because of its characteristics the protein is useful in the bread-making industry because it is able to trap carbon dioxide that is released in the flour-yeast reaction giving flour the chewiness characteristics and making it rise to keep a particular shape. There are different techniques of studying gluten components and analysing their characteristics. These include R5 ELISA test kits, biosensors, Quantitative Real-Time PCR, Liquid Chromatography Tandem Mass Spectrometry, Gel- Perme ation Chromatography and Size Exclusion High Performance Liquid Chromatography. These techniques however vary in terms of sensitivity and in the particular characteristics of gluten protein that they measure. Sandwich R5 ELISA Test Gluten protein components are mainly analysed using the sandwich R5 ELISA format. This analysis technique has a high sensitivity and it is quite specific for each gluten protein (Hochegger et al, 2015). Usually, sandwich R5 ELISA method is utilized in quantifying antigens especially when they are lowly concentrated and also when they are in a sample that has a larger amount of proteins which are non-gluten (Tranquet et al, 2012). It is based mainly on the R5 antibody, and therefore uses both R5 antibody and R5 conjugated antibody, which bind to different antigen sites. R5 antibody can recognize celiac epitopes that are potentially toxic repeatedly occurring in prolamins. These celiac epitopes include QQPFP, PQPFP, QQPYP, QQQFP, QLPYP and LQPFP. The epitopes are found in toxic-celiac peptides such as Gliadin 33 mer peptide, Gliadin 25 mer peptide, and the Gliadin 26 mer peptide (Guerdrum, 2013). The R5 ELISA assay has a gliadins quantification limit of 1.56 ppm. The assay must however be combined with the cocktail extraction solution. Several studies indicate that the R5 ELISA technique has been internationally accepted to be the main gluten detection method in foods by the Codex Alimentarius Commission. However, quantifying gluten in hydrolysed foods using this approach is slightly inaccurate since there is always need for two intact epitopes to help quantify gluten content (Hochegger et al, 2015). As a result, a competitive R5 ELISA that is based on an R5 monoclonal antibody is used to give a precise quantification. The later can quantify intact and/or fragmented gluten since the technique uses just one antibody and thus just one epitope is needed for complete gluten detection. Additionally, the competitive system is considered cheaper but faster as compared to the sandwich ELISA14 system (Guerdrum, 2013). According to the Codex Alimentarius Commission, there is need to modify R5 competitive ELISA assay in order to enable hydrolysed gluten detection. While the competitive technique is not compatible with the coc ktail extraction solution; a combination of the competitive assay and UPEX solution gives complete and very accurate gluten analysis. In regard testing for gluten in wheat, barley and rye R-5 antibody ELISA technique remains the favorite approach (Guerdrum, 2013). This is because the antibody neither reacts with glutenins, nor glutelins in rye and barley. Currently, the Codex committee on protein analytical methods recommends R-5 ELISA technique for analyzing gluten content in foods. Figure 1: obtained from https://www.wgpat.com/proceeding_25th.html The sandwich Omega analytical method thus varies from the competitive sandwich R5 ELISA test. The table below shows the variation in these methods in regard to detecting and characterising gluten protein. Figure 1 above shows a bar graph of gluten levels (ppm) in samples of buckwheat detected by Sandwich R5 ELISA test just after extracting using the cocktail solution. comparison of Sandwich Omega ELISA and the Competitive Sandwich R5 ELISA Sandwich Omega ELISA(Sandwich R5 ELISA) The Competitive Sandwich R5 ELISA It uses 40% of ethanol solution in extracting proteins Needs 2 epitopes for two antibodies to bind a protein It underestimates content of protein in particularly for hydrolysed and/or partially hydrolysed barley. Is able to detect heated i.e. denatured and also the unheated proteins even at gluten levels150 ppm Has higher sensitivity versions for protein analysis Detects gliadin, a gluten component Utilise extraction mixture of provided compounds Only needs a single but specific epitope i.e. the R5 monoclonal antibody Needs just one antibody Can detect protein that is highly heat resistant and toxic at the same time, very accurately The technique overestimates the content of protein in barley It cannot be used to determine hydrolysed gluten proteins It is able to detect heated and/or unheated proteins It can recognize barley, rye and wheat gliadin with 3 ppm and detection levels and can even measure5 ppm (Hochegger et al, 2015). The technique detects gliadin, one of the components of gluten. Figure 2: A comparison of Sandwich Omega ELISA and the Competitive Sandwich R5 ELISA Biosensors There are several biosensors that are used in the detection and characterization of gluten components. Biosensors are used especially to detect gliadin presence in in gluten-free food products. The first electrochemical biosensor relies on an antibody that is raised against gliadins immunodominant epitope that has 5.5 g/L of limit of detection (Gomes de Sousa Filho et al, 2014). The second biosensor is based on anti-gliadin Fab fragments adsorption properties on gold surfaces. The limit of detection for gliadin has been assessed by impedance and shown to be 0.42 mg/L and by amperometry that showed 3.29 g/L LOD (Nirantar et al, 2014). Lately, a biosensor referred to as quartz crystal microbalance biosensor which incorporates nanoparticles of gold can detect gliadin that has 8 g/Kg60 limit of detection (Sensors and Materials, 2016). A different biosensor which uses antibody-conjugated immunomagnetic beads of anti-gliadin and/or fluorescence-dye-loaded immunoliposomal nanovesicles so as to form a sandwich, registered 0.6 mg/L limit of detection for gliadin. Quantitative Real-Time PCR Polymerase chain reaction techniques can also be used to detect, characterize and even quantify DNA of cereals containing gluten. These techniques are able to not only characterize but also can provide various cultivars and enable the selection of genotypes coding for different gluten proteins that have the best quality of making bread (Debnath et al, 2009). PCR in combination with agarose gels can be used detect the presence of wheat in oats. In other studies quantitative PCR system in combination with the agarose gels is used to detect wheat, rye and barley contamination simultaneously, in gluten-free food products. The use of agarose gels is however disadvantageous and therefore the wheat DNA detection and quantification relies purely on quantitative polymerase chain reaction (Q-PCR) (Wang et al, 2010). There is currently a Q-PCR system that is reliable and provides rapid wheat DNA quantification in gluten-free food products and even in the raw materials. Its development is based SYBR Green I fluorescent dye and the modified extraction protocol of Guanidine-HCl or Proteinase K DNA. This particular system is very specific in nature and has a high sensitivity presenting up to 20 pg DNA/mg quantification limit (Debnath et al, 2009). This system when compared with the levels of glutens prolamin that have been determined using R5 ELISA commercial technique, it is clear that except for a few foods that are hydrolysed and/or highly processed; every sample that has prolamin levels that are above the quantification limit of R5 ELISA test (1.5 mg/kg) gives positive signals on Q-PCR system (Meral, 2016). It is therefore recommended that this particular Q-PCR system can be relied on to confirm the presence of gluten or generally wheat in foods, as a non-immunological tool, through the DNA pathway. This is especially useful in approving foods for celiacs and for those that have gluten allergy. Liquid Chromatography Tandem Mass Spectrometry The LC-MS/MS technique is also one of the methods for studying gluten proteins. The technique can be able to detect various species based on several markers with several confirmation points. This makes it less vulnerable to provide false positives and/or false negatives while giving far more detection confirmation (Lock, 2013). Since the LC-MS/MS technique is very specific in nature, it can distinguish species through the use of multiple peptide markers. LC-MS/MS mainly differs from R5 ELISA technique since it can rely on specific markers in wheat, oats, and rye. The individual markers for the above species can distinguish one from the other through LC-MS/MS. The LC-MS/MS tool is thus advantageous than the R5 ELISA technique in that it can use a number of peptide markers with a similar number of MRMs for every gluten peptide to detect gluten presence in a particular sample with different species (Liang et al, 2011). It is therefore the most feasible approach in detecting gluten in fo ods. It has also been proven that ELISA test kits fail to detect allergens in processed foods because of processing which brings changes on the proteins structure and thus preventing the binding of an antibody; leading to false negatives The LC-MS/MS technique detects 510 ppm gluten levels in gluten-free foods provides a linear response that is linear and also larger when compared to the response which is usually obtained ELISA tests(Liang et al, 2011). The current LC-MS/MS technique involves 80-fold sample dilution; that is then injected onto LC-MSMS system. This enables the system to potentially detect low gluten levels (0.55 ppm) particularly when the concentrate is collected and the peptide markers purified using under the SPE protocol (Creese Cooper, 2007). The presence of several markers for every variety of gluten and the ready availability of scans of MRM triggered product ion gives many gluten contamination confirmation points. This also provides confidence in gluten detection results obtained while reducing false positive and/or false negative risks that emerge in ELISA tests. Gel-Permeation Chromatography Size Exclusion High Performance Liquid Chromatography Gel-permeation chromatography relies on the principle of using materials that contain dextran to separate different macromolecules according to their molecular size differences. The procedure basically determines protein molecular weights and can also be used in decreasing concentration of salt in solutions of proteins (Bacskay et al, 2014). The gel- permeation column has a stationary phase which consist small-pored inert molecules. A solution containing various molecules that have varying dimensions, are continuously passed through the column at a flow rate that is constant. Molecules which are larger than the pore sizes are not able to permeate into the gel particles but retained within a restricted area between particles. The larger molecules can pass through the spaces that are between the porous particles to rapidly move inside the column (Jannah, 2015). On the other hand, smaller molecules diffuse into the pores as they get smaller, these molecules leave this column but with a longer retention time. The column material which is frequently used in gel permeation chromatography columns is the Sephadeks G type. Other column materials in this technique include agorose, dextran and polyacrylamide. Studies have indicated that conventional SE chromatography as described above has a lot of disadvantages as it is slow and usually has column beds that are unstable. Thus, results are likely to be unreliable and difficult to quantitate. As a result, there are Size-exclusion High Performance Liquid Chromatography (SE-HPLC) columns which are not only stable but uniform and reliable and thus are utilized in analysing proteins in cereals including gluten (Bacskay et al, 2014). The SE-HPLC separations are usually fast and this allows larger sample numbers for analysis. SE-HPLC has over time been utilized extensively in analysing cereal proteins. The SE-HPLC supports have different and/or varying porosity just like the conventional gel approach. An example of the Toya-Soda-made columns includes the TSK-4000SW, the TSK-3000SW and further, the TSK-2000SW columns. These columns have approximately 450, 240 and 130 A of pore size respectively (Creese Cooper, 2007). They are thus very useful in analysing high, intermediate and even low molecular weighted proteins. The process uses columns of measure 500 X 7.5 mm i.d. with 7-8 ml exclusion volumes and approximately 22 ml of total exclusion volume. Along the process, a guard column for TSK-3000SW with measures of 100 X 7.5 mm i.d. is used (Bacskay et al, 2014). A common solvent used in SE-H PLC particularly for cereal proteins like gluten; is 0.1 M sodium phosphate, with a pH of 7.0 and contains 0.1% SDS. The SDS in the solvent binds to proteins and solubilizes them in a way that the resultant protein-SDS complex sizes relate to protein MW. In the SDS presence, TSK-4000SW separates 10,000 to 1,000,000 MW proteins while TSK-3000SW usually separates 10,000 to 200,000 MW proteins (Tranquet et al, 2012). For proteins and/or peptides that have low molecular weight, the TSK-2000SW column is used. The flow rates in the columns are maintained at 1.0 ml/minute while the analyses carried out at room temperature. In the final analyses , the log MW plotted against the time of elution gives a straight line. Its equation is then used to approximate the MW of any other unknown proteins (Taddei et al, 2013). Usually, the standard measures are run on a daily basis and they are used to inform computer program updates on elution times, percentages, MW for every peak among others. Analysis of Gliadins using the SE HPLC Whole gliadin sample when tested shows the major peak for , and gliadins that correspond to 28,000 Molecular Weight. It also shows smaller peaks and/or shoulders which correspond 11000MW, 41000MW, 63000MW of ? gliadins and further 105000 MW high-molecular-weight gliadin (Tranquet et al, 2012). SE-HPLC is also used in comparing gluten proteins in native and/or reduced forms. For example, the high-MW gliadin is made up of intermediate MW heterogeneous oligomers. Its Molecular weight lies between gliadin and glutenin: and has an average MW of about 125,000 (Bacskay et al, 2014). The SE-HPLC technique rapidly confirms that the high-MW gliadin protein consists mainly, subunits of low-MW glutenin which are joined by way of disulphide bonds. SE-H PLC technique is useful in making comparison between proteins whether in their native or reduced states. Conclusion Gluten protein is a major requirement in bread making. Its levels must however be monitored especially understanding that it can affect a section of the population that are susceptible to celiac disease and those that have gluten allergy. This discussion highlights each technique that is relied upon to detect the amounts of gluten components in foods for rectification purposes. International companies that also sell wheat, barley, oat, rice and rye products rely on the technologies discussed to ensure that their products are gluten free. All the techniques discussed including sandwich R5 ELISA test kits, biosensors, Quantitative Real-Time PCR, Liquid Chromatography Tandem Mass Spectrometry, Gel- Permeation Chromatography and Size Exclusion High Performance Liquid Chromatography; is crucial in the food production sector. References Bacskay, I., Sepsey, A., Felinger, A. (2014). Determination of the pore size distribution of high-performance liquid chromatography stationary phases via inverse size exclusion chromatography.Journal of Chromatography A,1339, 110-117. Creese, A., Cooper, H. (2007). Liquid chromatography electron capture dissociation tandem mass spectrometry (LC-ECD-MS/MS) versus liquid chromatography collision-induced dissociation tandem mass spectrometry (LC-CID-MS/MS) for the identification of proteins.Journal of the American Society for Mass Spectrometry,18(5), 891-897. Debnath, J., Martin, A., Gowda, L. (2009). A polymerase chain reaction directed to detect wheat glutenin: Implications for gluten-free labelling.Food Research International,42(7), 782-787. Development of Portable Quartz Crystal Microbalance for Biosensor Applications. (2016).Sensors and Materials, 1. Gomes de Sousa Filho, J., Wiersma, C., Timpe, S., Doyle, B. (2014). Detection of interactions between botanical extracts and protein using a quartz crystal microbalance biosensor.Planta Medica,80(10). Guerdrum. (2013). The Determination of Gluten using the R5 Competitive ELISA Method.Journal of the American Society of Brewing Chemists. Hochegger, R., Mayer, W., Prochaska, M. (2015). Comparison of R5 and G12 Antibody-Based ELISA Used for the Determination of the Gluten Content in Official Food Samples.Foods,4(4), 654-664. Jannah, A. (2015). Isolation and Characterization of Rice Bran Protein Using NaOH Solution.ALCHEMY,4(1). Lacorn, M., Weiss, T. (2015). Partially Hydrolyzed Gluten in Fermented Cereal-Based Products by R5 Competitive ELISA: Collaborative Study, First Action 2015.05.Journal of AOAC International,98(5), 1346-1354. Lexhaller, B., Tompos, C., Scherf, K. (2017). Fundamental study on reactivities of gluten protein types from wheat, rye and barley with five sandwiches ELISA test kits.Food Chemistry,237, 320-330. LIANG, Y., WU, C., DAI, Z., LIANG, Z., LIANG, Z., ZHANG, L., ZHANG, Y. (2011). Microchip-based reversed-phase liquid chromatography-tandem mass spectrometry platform for protein analysis.Chinese Journal Of Chromatography,29(6), 469-474. Lock, S. (2013). Gluten Detection and Speciation by Liquid Chromatography Mass Spectrometry (LC-MS/MS).Foods,3(1), 13-29. Meral, R. (2016). Polymerase chain reaction (PCR) for the detection of gluten.Journal Of Biotechnology,231, S54. Miranda-Castro, R., de-los-Santos-lvarez, N., Miranda-Ordieres, A., Lobo-Castan, M. (2016). Harnessing Aptamers to Overcome Challenges in Gluten Detection.Biosensors,6(2), 16. Nirantar, S., Li, X., Siau, J., Ghadessy, F. (2014). Rapid screening of proteinprotein interaction inhibitors using the protease exclusion assay.Biosensors And Bioelectronics,56, 250-257. Nordqvist, P., Thedjil, D., Khosravi, S., Lawther, M., Malmstrm, E., Khabbaz, F. (2011). Wheat gluten fractions as wood adhesives-glutenins versus gliadins.Journal Of Applied Polymer Science,123(3), 1530-1538. Olexov, L., Dovi?ovi?ov, L., vec, M., Siekel, P., Kuchta, T. (2006). Detection of gluten-containing cereals in flours and gluten-free bakery products by polymerase chain reaction.Food Control,17(3), 234-237. Taddei, P., Zanna, N., Tozzi, S. (2013). Raman characterization of the interactions between gliadins and anthocyanins.Journal Of Raman Spectroscopy,44(10), 1435-1439. Tranquet, O., Larr, C., Denery-Papini, S. (2012). Selection of a monoclonal antibody for detection of gliadins and glutenins: A step towards reliable gluten quantification.Journal Of Cereal Science,56(3), 760-763. WANG, H., CHEN, R., CHEN, P. (2010). Detection of Genetically Modified Herbicidetolerant Crops by Real-time Fluorescent Quantitative PCR Assay*.Chinese Journal Of Appplied Environmental Biology,2009(6), 866-870.

Board Composition and Operational Risk †MyAssignmenthelp.com

Question: Discuss about the Board Composition and Operational Risk. Answer: Introduction The report helps in understanding about the different kinds of operational risks that are involved in a company. The company that has been taken in this particular report is HG Metal Manufacturing Company that is situated in Singapore. An operations manager is appointed in the respective company as to properly assess the risk and suggest as well as implement different measures to reduce such operational risks from the company. The risk management approaches and framework of HG Metal Manufacturing Company has to be properly analyzed as well. The main aim of the report is to understand as well as analyze the different incidents in the past that has occurred as the operational risks in the respective company as well as the industry. The main purpose of the report is to analyze the good practices of the safety technologies with proper tools and techniques as well. The impact of the hazardous substances treatment has to be implemented as this will help in treating the risk. The structure of the report will include the proper process of treatment of the different kinds of risk and this will help in reviewing the good practices that will help in reducing such operational risks from the respective company. The impact has to be properly discussed that will affect the company and recommendation has to be properly implemented by the Operations Manager as this will help in assessing the risk that can be a danger to the entire company. HG Metal Manufacturing Limitedis a metal company that was founded in the year 1971 that is the retailer of steel as well as metal products. The respective company provides end to end solution to the customers that range from services relates to distribution to the downstream along with the value added services. The HG Metal Manufacturing LimitedCompany is properly equipped with different kinds of sourcing capability that is strong in nature and they have a huge network of suppliers from different countries namely Japan, Russia, Tokyo along with other European countries. The nature of the business is that the company has also offered steel and metal solutions that are customized in nature for different industries namely transportation, energy as well as electronics. Structure of the risk management team The main duty of the risk managers is to analyze the safety of the supervision along with proper handling of different claims. The operations manager has to report to the higher officials in the management regarding any kind of losses, other programs related to prevention of loss of the company as well as results of the insurance marketing. Proper identification of the exposures related to exposure has to be ascertained as this will help them in identification of the solutions of the insurance related products. The entire team will work effectively to minimize the payment of the loss for litigation as well as claims. The operational management of risk has focused previously on the mitigation of the risks from the internal processes that are internal in nature but due to the increase in the regulatory requirements along with the requirement of the improvement in the performance of the business the scope of Operations Risk Management has been expanded. The ORM program of the respective company is based on framework of the sponsoring company and this will help their clients in reimagining the different risks as well as compliance process with internal control that will offer: Reliable reporting that is financial in nature Proper safeguarding of assets (Armstrong and Taylor 2014) Efficient and effective programs Proper compliance with regulations and law Risk Management Team This is the duty of the operations manager that all the workings that are manufactured in cost effective as well as timely manner as this will help in meeting the quality requirements Proper improvement of the operational systems along with processes is the other duty that guarantees the well being of the entire organization and ensure the efficiency at the warehouse Proper examination of the financial statements as this will be used to improve the profitability Proper contribution towards the achievement of the operational as well as strategic objectives Lastly, it is the duty of the operational manager to cater to the concerns of the different personnel (McNeil, Frey and Embrechts 2015) These are the different activities that have to be performed by the operations manager of the respective company HG Metal Manufacturing Limitedwherein they have to handle and manage the raw materials as well as personnel. The proper oversight of the entire inventory along with supplies as well as purchasing has to be properly taken care by the operations manager. The operations manager has to analyze the kind of operational risk that is affecting the brand image of the company and in this particular scenario, the change in the organization can come in different forms that can affect the structure of the entire business. Review of Past incidents In HG Metal Manufacturing LimitedCompany, there was a risk that has been faced by the respective company due to the low quality of the products that are produced by them. In the past, there was a risk related to the poor quality of the metal products that are sold by HG Metal Manufacturing LimitedCompany and the customers faced lot of difficulties as well. The proper quality assurance is essential for the company to provide to the customers as the customers trust the brands and they go ahead with the purchase of different goods as well as different services. HG Metal Manufacturing LimitedCompany supplied different metal and steel products that were sold to the customers but they were of low quality as the assurance of quality was not provided to the clients and this created bad reputation for the defective products that were sold by the company. There were different kind of defects in the steel and metal product and there is a policy of replacement that has been provided by the compa ny to the customers when the problem is discovered (Aven 2016). According to Farrell and Gallagher (2015), operations risk is the kind of issue that affects the internal ability of the company to supply as well as produce different goods as well as services. There can be different risks that can affect the reputation of the firm as well as the impression upon the customers as well. In the last few decades, Barakat and Hussainey (2013) commented that risk management in the supply chain management has properly developed but there are different operational risks in the different companies. Chiu and Choi (2016) outlined that the risk management is the area with different conflicting terms along with proper critical reflection on the principles as well as regulations (Chen, Sohal and Prajogo 2013). The operational risks that has been faced by Reliance Company as the gap analysis was done with the help of proper vulnerability assessment. The root cause analysis was properly reviewed as this will help in resolving such issues with proper implementation of different strategies as shown in the picture below: Identification of hazard risk There are different methods of the identification of the risks by HG Metal Manufacturing LimitedCompany on the basis of the databases of the different events that has happened in the past as this will be helpful in nature to understand such issues that has occurred in the future and try to resolve such issues as this will help in understanding the potential risks that are involved in such kind of cases (Thomas, Crook and Edelman 2017). HG Metal Manufacturing LimitedCompany has to identify the root causes of the different operational risks wherein proper identification of the undesirable events that has gone wrong and proper identification of the potential impacts is essential as well in such events. Secondly, it is essential for the company to identify the different functions that are essential in nature and possible modes have to be identified that are crucial for gaining success (Xu et al. 2014). The method of the qualitative risk assessment is essential to be done by HG Metal Manufacturing LimitedCompany wherein the following steps have to be followed: Screening of the risk is essential to be done by the respective company as this will help in identification of the different risks that are operational in nature. The operational risks has to be properly omitted by the company as this will help the company along with the employees of the organization to understand the causes of the operational risks and resolving such issues with proper implementation of different strategies as well. It is determined with proper analysis of: High impact and low probability (Lam 2014) Low impact and high probability Low impact and low probability High impact and high probability (Soin and Collier 2013) Pareto diagram helps in identification of the different operational as well as other risks with proper implementation of the different strategies that will help in mitigating the risks and this helps in improving the brand name of the organization as well. Analysis of the failure modes as well as effects helps in assessing the risk of the organization that has caused failure in the organization as well. The failures can be related to the operational risks of the company wherein there is impact on the demand as well as supply of the goods and services (Wiengarten et al. 2016). There are two different tools and techniques that help in identification as well as analysis of the operational risks of HG Metal Manufacturing LimitedCompany are as follows: Information gathering technique is one such technique that helps in rating the risk that is high or low in nature. The respective company has to understand whether the risk is dangerous, then the company should immediate stop such process as this will help in implementation of the control techniques. The proper information from the past records has to be collected by the operations manager in the organization as this will help in emphasizing the requirements that are essential in nature to reduce such risks with the help of proper protection. Root Cause Analysis is another technique wherein the level of the risk has to be identified and the involvement of such risks as well. This will help in determination of the risks that are hazardous in nature and such risks have to be controlled with reviewing and monitoring the hazards that can affect at the workplace. There are different kinds of documentation required for the proper assessment of the risks in the workplace that are as follows: Level of the risk that is involved (Heckmann, Comes and Nickel 2015) The requirements that are legislative in nature Implementation of different kinds of control measures that will help in reducing the operational risks in the company Reviewing as well as monitoring the hazards at the workplace as this will help in reducing the kind of risks and this will help in measuring the suitable risk factor as well. These are the different kinds of tools and techniques as this will help them in understanding the level of risk and implement the safety technologies with the help of proper advancement of technologies. Risk evaluation and analysis There are different safety control technologies that will help HG Metal Manufacturing LimitedCompany in order to reduce the operational risks in the respective company. The analysis can be done with the help of the following the steps that are as follows: Proper segregation of the task as this will help in reducing the risks related to theft as well as fraud. This will help in proper prevention of taking proper advantage of the different numerous aspects of business practices as well as transactions. Secondly, the risks has to be curtailed in the process of the business as this will help in reducing the complexities in the business and this will help in mitigating the operational risks. The respective organization can achieve and reduce the complexities with curtailing the different manual activities along with proper implementation of the processes of business (Therivel and Paridario 2013). The respective organization named HG Metal Manufacturing LimitedCompany has to create a proper ethics that is related with proper mitigation of the operational risks management. The ethics of the organization has to be properly enforced by proper combination of the different workplace principles along with the personal values as well The respective organization needs to allocate the right individuals for the right kind of tasks as this will help in reducing the issues that pertains to the execution of the entire process of the business along with the proper skills as well as usage of advanced technologies (Adeusi et al. 2014). This will help in resulting proper and appropriate utilization of the workforce as well as proper adherence to the technologies as well as timelines with proper enhancement of the technologies as this will help in reducing the number of errors as well as breakdown of the processes. Proper monitoring as well as evaluation at proper intervals is essential and necessary in nature as the process of the business is effective as well as efficient in nature and this will help HG Metal Manufacturing LimitedCompany in reducing the operational risks from the different areas of the business (Hopkin 2017). The well designed indicators of the performance have to be properly implemented by the company as this will help in enhancing the process of business. The key performance indicators are critical in nature as this will help in making timely decisions as well as proper mitigation of the different operational risks that can impact the performance of the entire business. The continuous monitoring as well as reviewing is essential in nature as this will help in identification of the discrepancies proactively as well as manage them accordingly (Sharifi et al. 2016). The assessment of the risk has to be done periodically in the company HG Metal Manufacturing LimitedCompany as this will provide proper relief to the entire organization and this will help in making the regulatory frameworks for the entire organization to mitigate such operational risks in the organization. It is imperative to be properly risk ready by properly gauging the regulatory obligations along with the different competencies of the skills of information technologies as well (Alexander 2013). The risk incidents as well as different activities those are remedial in nature for proper effectiveness in the entire organization. The strategies that are effective in nature will help in countering the future risks. The risk occurrences that has happened previously has to be properly analyzed by the company in order to make some amendments in the future as well as in the present as this will help in handling the current operating scenario at the workplace. The task in hand that is critical in nature is wherein the organization can implement different the steps in reducing such kinds of risk. Proper corporate governance as well as the compliance platform enabled proper advancement of technologies and this helped in effectively support the proper implementation of the 7 step approach to reduce the operational risk management (Hora and Klassen 2013). This will help the respective company HG Metal Manufacturing LimitedCompany in handling such risks and this helped them in maintaining and this will help in mitigating the operational risks and this helped the entire company along with the employees to support proper implementation of different technologies (Sturm 2013). Review of proper good practices and safety technologies There are different advantages as well as disadvantages in the risk treatment of the operational risk in the respective company wherein the risks can be nurtured and with the proper handling of the previous risks, the future risks can be mitigated with proper implementation of technologies. However, on the other hand, there are disadvantages as well wherein there can be unmanaged losses that can impact the entire business and the external entities along with ambiguity that is uncontrollable in nature (Wang and Hsu 2013). The potential threats cannot be managed properly as there are external uncertainties that cannot be controlled by the management. Recommendation Therefore, it can be recommended that the respective company has to adopt certain other strategies in mitigating the different kinds of operational risks and this will help in focusing properly on the assessment of the risks and this will help in understanding the impact of the risk on the entire business as well. This will help in promoting the culture of the entire organization as well. Lastly, the risks can be nurtured properly by compensating the risks within rigid comparisons along with different choices that will help in mitigating the risks as well from the entire business. These are the different process through the advancement in technologies that will help in mitigating the operational risks. References Adeusi, S.O., Akeke, N.I., Adebisi, O.S. and Oladunjoye, O., 2014. Risk management and financial performance of banks in Nigeria.Risk Management,6(31). Alexander, K. ed., 2013.Facilities management: theory and practice. Routledge. Armstrong, M. and Taylor, S., 2014. Armstrong's handbook of human resource management practice. Kogan Page Publishers. Aven, T., 2016. Risk assessment and risk management: Review of recent advances on their foundation.European Journal of Operational Research,253(1), pp.1-13. Barakat, A. and Hussainey, K., 2013. Bank governance, regulation, supervision, and risk reporting: Evidence from operational risk disclosures in European banks.International Review of Financial Analysis,30, pp.254-273. Bromiley, P., McShane, M., Nair, A. and Rustambekov, E., 2015. Enterprise risk management: Review, critique, and research directions.Long range planning,48(4), pp.265-276. (Bromiley et al. 2015) Chen, J., Sohal, A.S. and Prajogo, D.I., 2013. Supply chain operational risk mitigation: a collaborative approach.International Journal of Production Research,51(7), pp.2186-2199. Chiu, C.H. and Choi, T.M., 2016. Supply chain risk analysis with mean-variance models: a technical review.Annals of Operations Research,240(2), pp.489-507. Farrell, M. and Gallagher, R., 2015. The valuation implications of enterprise risk management maturity.Journal of Risk and Insurance,82(3), pp.625-657. Heckmann, I., Comes, T. and Nickel, S., 2015. A critical review on supply chain riskDefinition, measure and modeling.Omega,52, pp.119-132. Hopkin, P., 2017.Fundamentals of risk management: understanding, evaluating and implementing effective risk management. Kogan Page Publishers. Hora, M. and Klassen, R.D., 2013. Learning from others misfortune: Factors influencing knowledge acquisition to reduce operational risk.Journal of Operations Management,31(1), pp.52-61. Lam, J., 2014.Enterprise risk management: from incentives to controls. John Wiley Sons. McNeil, A.J., Frey, R. and Embrechts, P., 2015.Quantitative risk management: Concepts, techniques and tools. Princeton university press. Sharifi, S., Sharifi, S., Haldar, A., Haldar, A., Rao, S.N. and Rao, S.N., 2016. Relationship between operational risk management, size, and ownership of Indian banks.Managerial Finance,42(10), pp.930-942. Soin, K. and Collier, P., 2013. Risk and risk management in management accounting and control. Sturm, P., 2013. Operational and reputational risk in the European banking industry: The market reaction to operational risk events.Journal of Economic Behavior Organization,85, pp.191-206. Therivel, R. and Paridario, M.R., 2013.The practice of strategic environmental assessment. Routledge. Thomas, L., Crook, J. and Edelman, D., 2017.Credit scoring and its applications. Society for industrial and Applied Mathematics. Wang, T. and Hsu, C., 2013. Board composition and operational risk events of financial institutions.Journal of Banking Finance,37(6), pp.2042-2051. Wiengarten, F., Humphreys, P., Gimenez, C. and McIvor, R., 2016. Risk, risk management practices, and the success of supply chain integration.International Journal of Production Economics,171, pp.361-370. Xu, G., Dan, B., Zhang, X. and Liu, C., 2014. Coordinating a dual-channel supply chain with risk-averse under a two-way revenue sharing contract.International Journal of Production Economics,147, pp.171-179.

Legalization Of Marijuana Essays (186 words) - Cannabis,

Legalization Of Marijuana Legalization of medical marijuana This act may be cited as the " legalization of marijuana" Sec. 1 This bill will be for the well being of the pain stricken patients in our nation's hospitals, a doctor that will decide if a patient is illegible to receive marijuana. Sec. 2 Patients will only be permitted to get their marijuana at a hospital or a licensed distributor. Sec. 3 The patient's will have to buy the marijuana at a discount from the hospital, or distributor using their own money or if they're health care will cover the treatment. The discount will be 1/8 of the current street price on pure marijuana. The government will grow their own marijuana in a dis-closed location for security reasons. Sec.4 The Federal Drug Administration will monitor the distribution and handling of the medical marijuana Sec.5 This law will take affect 1 year after it is ratified due to the long periods of time needed to locate, distribute and grow this marijuana. Case that needs this treatment now will be allowed the first dosage that becomes available. Bibliography www.Infoseek.com/top ten killers in the United States www.yahoo.com/me-marijuana www.yahoo.com/marijuana

Georgia State Unit Study - Geography, State Symbols Facts

Georgia State Unit Study - Geography, State Symbols Facts These state unit studies are designed to help children learn the geography of the United States and learn factual information about every state. These studies are great for children in the public and private education system as well as homeschooled children. Print the United States Map and color each state as you study it. Keep map at the front of your notebook for use with each state. Print the State Information Sheet and fill in the information as you find it. Print the Georgia State Map and fill in the state capital, large cities and state attractions that you find. Answer the following questions on lined paper in complete sentences. State Capital What is the capital?State Flag What is in the circle of stars?State Flower Who approved the state flower in 1916?State Crop Georgia produces what percentage of the nations supply?State Fruit This fruit gives the state its nickname - what is it?State Bird What is the state bird? Coloring PageState Marine Mammal How long does this mammal grow?State Fish What is the state fish?State Tree What is the state tree?State Insect How does this insect help Georgias economy?State Butterfly What is the coloring of this butterfly?State Vegetable What is unique about this vegetable?State Song Who wrote the state song?State Seal What do the three pillars stand for? Coloring PageState Motto What is the state motto? Georgia Printable Pages - Learn more about Georgia with these printable worksheets and coloring pages. Georgia Word Search - Find the Georgia State Symbols. Did You Know... List two interesting facts. Seven Natural Wonders of Georgia - Most people have heard of the seven wonders of the world. Not as many have heard of the seven natural wonders in the state of Georgia. The Childrens Museum of Atlanta - Take a virtual tour. From Zoo Atlanta: The Animals; Panda Mask; Meerkat Maze Georgia History 101 - An overview of Georgia history. The King Center - Learn all about Dr. Martin Luther King, Jr. Savannah River Ecology Laboratory - Meet the reptiles and amphibians that call the Savannah River region their home. Georgia Flag Printout - Learn about Georgias new flag. Georgia Map/Quiz Printout - Can you answer the questions about Georgia? Odd Georgia Law: No one may carry an ice cream cone in their back pocket if it is Sunday. Related Resources: More State StudiesGeorgia History and Activity BooksHands-on GeographyHands-on Geography Activity Books Additional Resource: Introducing the email course Our 50 Great States! From Delaware to Hawaii, learn about all 50 states in the order they were admitted to the Union. At the end of 25 weeks (2 states per week), youll have a United States Notebook filled with information about each state; and, if youre up the the challenge, you will try recipes from all 50 states. Will you join me on the journey?

Electric Field Simulation Essay Example | Topics and Well Written Essays - 2500 words

Electric Field Simulation - Essay Example Through COMSOL Multiphysics we discovered the results after simulating an electric field by using 2D and 3D of the electrostatic module. These modules provided many kinds of movement in the electric field for the three electrodes, which were energised with +1V, 0V, and -1V electrical voltages. Also, the strong and weak points are posted between the three electrodes and show the electric potential of the field. Finally, this paper will show the form of the distribution of electric potential and electric field between the three electrodes for the above mentioned conditions. Introduction: After Michael Faraday discovered the electric field, he developed electricity into something practical that could be used in many technologies, especially microsystem devices. According to James Clear Maxwell, â€Å"the portion of space in the neighborhood of electrified bodies† is called an electric field [1]. At present, there are several applications used that are related to microtechnology a nd are beneficial to our lives. A good application in medicine is biology cells within medical laboratories. In this way, application is applied to the electric field to move cells and separate or analyze cells via impact electric forces. The movement, separation, and analyzing is done through a technique known as AC electrokinetics. This technique occurs when an electric field interacts with dipoles, but it depends on forces between repulsion and rotation by altering the nature of the dynamic field [2]. This new technique is beneficial in biotechnology because of the electric field [3]. Also, the AC electrokinetics technique depends on a delicate process known as dielectrophoresis. This is â€Å"the migration of uncharged particles towards the position of maximum field strength in a non-uniform electric field† [4]. The basic principle operation of dielectrophoresis is by deference of electromagnetic and dielectric properties. For example, the separation of cancer cells is be hind the electrodes, while the natural cells move away from the electrodes due to variations of the electric field [5]. Figure 1 shows the forces of attraction and repulsion between cancer cells and normal cells. Fig.1. Basic Principle of Dielectrophoresis An electric field is a region around a charged particle or object within which a force would be exerted on other charged particles or objects. It is defined as an area between two charges and then there is a force (positive or negative) exerted [6]. The forces exerted on the test charge will be directly proportional to another charge according to Coulomb’s law [7]: Fe ? q1 q2 If divide the forces on the test charge: E=Fe /q ' Where E = electric field (N/C) and F = force (N) and q' = charge on test charge (C) Also, according to Coulomb’s law, we can find E where: = the permittivity of free space Then we can calculate the electric flux by using Gausses’ law [8]: Q = ? E.d There is a relationship between the elec tric field and electric potential if the electric potential is identified in an action area, then we can calculate the value of the electric field by: dV = - E.d. However, the electric potential consists of lines called equipotential lines. There is a direct correlation between the electric field lines and the energy of electric potential because the first one always puts the electric potential of direction that causes dropping electric potential [9], whereas, in this case, we are dealing with an accelerometer that

Marketing Plan for HIV drug Essay Example | Topics and Well Written Essays - 3250 words

Marketing Plan for HIV drug - Essay Example Promotional aspect of marketing mix is usually considered to be marketing communication. It is all about conveying a common message across different media channels so as to ensure that it reaches target audience. Marketing communication is a strategic approach adopted by a company in order to reach target audience. In this study a product would be outlined which is losing significance in modern world. HIV or AIDS is regarded as a global problem. In its early years it was an incurable disease but in present scenario it is a health problem which can be effectively addressed. The percentage of death rate is considerably falling due to introduction of various treatment and retroviral drugs. However the problem is linked with lack of awareness program about HIV drug. Young people often do not remember about these drugs or are not determined to consume HIV drugs. Through this study a marketing campaign will be designed that could influence target segment to purchase HIV drug and prevent su ch diseases from spreading. HIV is a global issue that has contributed towards death percentage rise. Modern treatments had been introduced by government and healthcare agencies to prevent this disease. There are new drugs being introduced which can eradicate this kind of disease from its roots. HIV drugs are being developed at a faster rate and it is inclined towards saving lives of HIV victims. There are few issues associated with marketing of HIV drug. Firstly it has been observed that individuals are less likely to accept their disease. This in turn restricts drug makers or health care agencies to efficiently reach out to target audience. Death rate due to HIV aids have been decreasing over the years and negligence has been main cause for this issue. On the other hand, individuals who agree to purchase this drug at times are not able to afford such high priced drugs. Affordability is a major area of concern for HIV victims. There are individuals affected by HIV

DNA Extraction From Chicken Liver

DNA Extraction From Chicken Liver Deoxyribonucleic acid (DNA) is the hereditary material in humans and almost all other organisms. Nearly every cell in a persons body has the same DNA. Most DNA is located in the cell nucleus (where it is called nuclear DNA), but a small amount of DNA can also be found in the mitochondria (where it is called mitochondrial DNA or mtDNA). The information in DNA is stored as a code made up of four chemical bases: adenine (A), guanine (G), cytosine (C), and thymine (T). Human DNA consists of about 3 billion bases, and more than 99 percent of those bases are the same in all people. The order, or sequence, of these bases determines the information available for building and maintaining an organism, similar to the way in which letters of the alphabet appear in a certain order to form words and sentences. DNA bases pair up with each other, A with T and C with G, to form units called base pairs. Each base is also attached to a sugar molecule and a phosphate molecule. Together, a base, sugar, and phosphate are called a nucleotide. Nucleotides are arranged in two long strands that form a spiral called a double helix. The structure of the double helix is somewhat like a ladder, with the base pairs forming the ladders rungs and the sugar and phosphate molecules forming the vertical sidepieces of the ladder. An important property of DNA is that it can replicate, or make copies of itself. Each strand of DNA in the double helix can serve as a pattern for duplicating the sequence of bases. This is critical when cells divide because each new cell needs to have an exact copy of the DNA present in the old cell. The extraction of DNA from cells and its purification are of primary importance to the field of biotechnology and forensics. Extraction and purification of DNA are the first steps in the analysis and manipulation of DNA that allow scientists to detect genetic disorders, produce DNA fingerprints of individuals, and even create genetically engineered organisms that can produce beneficial products such as insulin, antibiotics, and hormones.ÂÂ   Once the DNA has been isolated, it is essential to accurately determine its concentration for subsequent manipulation such as cloning or sequence determination. To quantify the amount of DNA that extracted by using spectrophotometry. The aims of this experience is to: To use the properties of DNA to isolate long strands of DNA from liver cells. To determine the yield of DNA isolated from a given amount of tissue. To examine the light absorbing properties of purified DNA. To examne the relationship between the concentration of a DNA solution and the absorbnce at 595nm of DNA-diphenylamine solution. To generate a standrad curve relating DNA concentraton with the absorbance of DNA-diphenylamine solutions. To use a standard curve to determine the concentration of an unknown DNA solution. Materials and Methods As per lab manual. Results Firstly, the chicken liver cell homogenate is treated with a salt solution such as NaCl and a detergent solution containing the compound SDS (sodiumdodecyl sulfate). These solutions break down and emulsify the fat proteins that make up a cell membrane. Finally, ethanol is added because DNA is soluble in water. After adding ethanol a relatively clear aqueous will be produced, the first layer is the milky solution that is the aqueous phase with DNA, the middle layer is the solid (precipitate proteins). The bottom layer is a clear solution (organic). The DNA can be spooled (wound) on a stirring rod and pulled from the solution at this point. The amount of DNA solution we got is 5.4ml.Than we put the DNA solution in 2ml tube (1.041g). The total weight of DNA solution and tube is 1.106g. The amount of DNA we got is 1.106-1.041g = 0.065g. Next we prepare 4 standard tubes by adding TE buffer (ml) to the DNA standard solution (ml). And also added to each of the 3 samples of my DNA. The total DNA (mg) is recorded in the table 1. The observed colour change of 4 standard tube and my 3 samples are recorded in table 2 and 3. We pipette the DNA samples and each standards tubes into separate wells of a 96 well microtitre plate. We measured the absorbance at 595nm of the DNA-diphenylamine solutions using the plate reader. Our results are shown in the graph with the used of the reading of table 4. Form the graph we find that the concentration of undiluted DNA is 0.232=0.46mg/ml. Discussion and Conclusions For this experiment we determinate the yield of the DNA isolate from given amount of tissue is: 1g -> 63mg 0.065g -> 4.095mg (wet weight of the DNA to dry weight) 3ml -> 4.095mg 5.4ml -> 7.371mg (DNA in the entire aqueous phase is collected) 3. 4ml -> 7.371mg 5.3ml -> 9.767mg The final calculation of the dry DNA is 9.767mg/g liver. For the experiment we examine that the light absorbing properties of purified DNA. The wavelength is range 220-300nm. The wavelength of the DNA is 260nm. We also calculated that the yield of DNA per g of liver from Lab 2 is: The amount (mg) of DNA contain => 0.461.5=0.69mg Aqueous from lab 1 = 5.4mg 0.69/2 =0.345mg (0.3455.4)/3 = 0.621mg The final value in mg of dry DNA/g liver is: 0.621mg/g. In the end of the experiments, we managed to complete our objectives. In summary, we learn that the alcohol can causes DNA to precipitate, or settle out of the solution, leaving behind all the cellular components that arent soluble in alcohol. As alcohol is less dense than water, so it floats on top forming two separate layers. We also learn that the advantage of spectrophotometry is that diphenylamine only reacts with DNA more accurate as RNA would not be determined. The disadvantage of spectrophotometry is that it always requires standard solution. The advantage of calculating of yield by its weight is that it does not require standard solution. The disadvantage of calculating of yield by its weight is that it is less accurate as RNA is counted in.