Homogenized Mushroom Tissue Based Biosensors

Homogenized Mushroom Tissue Based Biosensors ABSTRACT Homogenized mushroom (Agaricus bisporus) tissue based biosensors by using plant tissue materials is a relatively new development in the biosensor technology. A simple in dip stick kind of visual ethanol biosensor that based on immobalised alcohol oxidase (AOX) homogenized from mushroom tissue onto polyaniline (PANI) film for ethanol (> 5% concentration ) detection in toiletries products. The colour going to change from green to blue due to response of biosensor to the ethanol and the change of colour can be seen by naked eyes. Regarding the enzymatic reaction of ethanol, acetaldehyde and hydrogen peroxide will produced, then PANI film is latter going to be oxidised. The method used to immobalise AOX onro the PANI film is by adsorption. Regarding immobilisation process, AOX solution need to deposit on the PANI film and then left it at room temperature within 30 minutes until it dry. Scan and analyse the changes film’s colour to obtain the biosensor’s response characteris tics toward the ethanol. The biosensor respond. Therefore, this simple visual biosensor is suitable for all-range-aged community to determine the safeness of certain toiletries products from the ethanol. Keywords : Biosensor; Alcohol Oxidase Ethanol; Mushroom Tissue; Polyaniline; Toiletries 6.0Â  EXPERIMENTAL 6.1Â  Chemicals 10-40 units/mg protein of Alcohol Oxidase or simply called (AOX) (A2404, EC1.1.3.13) which extract from mushroom (A. Bisporus). These mushroom can be bought at fresh market as culture vegetables. Before use make sure store it at 4Â °C. In order to immobalization use 225 bloom of gelatin from calf skin and 25.0% glutaraldehyde. Ascorbic acid, 2-propanol, D-glucose, n-butanol and many more chemicals needed can be purchased from Sigma, St. Louis, USA. Aniline with AR-grade, gallic acid (G7384) and l-cysteine (W326305) can be purchased from Sigma Aldrich (Saint Louis, MO, USA). Ethanol which contain >99.5%, methanol, orthophosphoric acid (85%) and sodium hydroxide (pellets) can be delivered by Merck (Nottingham, UK). All needed chemicals are from commercial source which in analytical grade. Millipore Direct-QTM 5 purification system provide the Milli-Q water. Prepare ethanol’s stock solutions in 0.1 M phosphate buffer at suitable pH daily and store it in refrigerator at 4 Â °C. For pH studies, 0.1 M of the phosphate buffer solutions with pH values between 4 and 8 were can be used and to measure the pH value, use commercial glass electrode and pH-meter (model 9318, Hanna Instruments, Woonsocket, RL, USA) and calibrate it at the pH values of 4.00, 7.00 and 9.00. 6.2Â  Homogenization of Mushroom Tissue Homogenize 200 mg mushroom tissue in the 400ml phosphate buffer. Mix this 300 ml homogenate and 10 mg gelatin. Incubate it for 5 minutes at 38Â °C to make sure the gelatin is dissolved. 6.3Â  Preparation of Polyaniline (PANI) Film In order to prevent bumping in the aniline, purify the aniline with vigorous and rapid stirring by distilled under vacuum. Prepare PANI dispersion as nanofibre and Huang and Karner (2006) already mention the method to should to be used to build it. Mix 3.2 mmol or 0.3 g of purified aniline with 10 mL of 1.0M Hydrochloric Acid (HCL). Mix 0.8 mmol or 0.18 g of Ammonium peroxydisulfate into another 10 mL aliquot acid solutions. Add aniline-acid to oxidant and these two solution will mix rapidly within 30 seconds and then allow it to react in undisturbed overnight condition. On the next day, wash the polyaniline by using water and centrifuged. Supernatant liquor with pH 3.3 and strong green colour will form and it is indicate as the PANI particles which can be observed after several times of washing. Any remaining particles with size larger than 1ÃŽ ¼m must be removed before casting by passing the dispersion through a 55-mm glass fiber filter (Whatman GFA, Kent, UK) which is attach to va cuum source. Cast directly PANI dispersion on a substrate of polystyrene and then, left the thin film of PANI that on the sheet of polystyrene in the dark to dry after cut it into individual in 10mm2 size. Next, store that ready film at 4Â °C. The thickness must be 0.7ÃŽ ¼m and use SEM images to determine it. To make sure the thickness of the film is always in the same magnitude order, it must be determined routinely. Then, choose 0.7 ÃŽ ¼m thickness of PANI film to use for the further experiment result in good of PANI film fabrication reproducibility. 6.4Â  Enzyme Immobilization Immerse the PANI film in 0.1 M phosphate buffer which have pH 7.0 to make sure the condition of PANI film is at natural condition which means at pH 7.0. After that, deposit appropriate concentration about 10 ÃŽ ¼L of AOX solution which is from homogenate of mushroom tissues and phosphate buffer on the PANI film and left it to dry within 30 minutes. For the further use, store this PANI film with immobilised AOX at 4 Â °C. 6.5Â  Biosensor Construction Construct a dip stick test visual biosensor of PANI film with immobilized AOX as Figure 1, connect AOX/PANI film with a handle which made by cellulose paper or can use transparent plastic tape. To way to use this this dip-stick format visual biosensor by just dipping this kind of biosensor into the toiletries sample solution for a several seconds (Â ±5 s), then the change of colour can be seen by baked eye if the concentration of ethanol is >5% since that only amount allowed to be in toiletries products and use image analysis for the quantitative measurement change of colour. (a)(b) Figure 1: (a) dip stick format of biosensor (b) dip biosensor into toiletries sample solution (Kuswandi et.al,2014) 6.6 Colour Change Recording See the change of colour by naked eye during alcohol detection since this biosensor is kind of visual mode. Use scanner for example Canon, Cano Scan, Japan and Tokyo for quantification of colour measurements. The presence of ethanol exceed 5% of concentration in toiletries sample solutions will change the colour of the biosensor from green to blue. The detection can be done by dipped the biosensor which in the form of dip stick test in the sample solutions in 5 seconds. Use ImageJ program which can be used as online applet, free download application or can be used in any computer together with Java 5 (Dougherty, 2009 Rueden et.al, 2007) in order to assess the colour after it has been scanned. The purpose s to determine the mean RGB colour value. 7.0Â  EXPECTED RESULT The expected result from this research is the colour of dip stick AOX/PANI film biosensor will change from green to blue if there presence of >5% of ethanol in toiletries products after the biosensor is dipped into the toiletries sample solutions for 5 seconds. Since this is the visual mode biosensor, the change of colour can easily seen by naked eyes. On the hand, use scanner (Canon, Cano Scan, Japan, Tokyo) for quantitative colour measurement and then use ImageJ program to assess the colour change of biosensor and to determine the mean RGB colour value. (Collins, 2007)

Cocaine and the Brain: The Neurobiology of Addiction Essay -- Biology

Cocaine and the Brain: The Neurobiology of Addiction In the eyes of the public, the word addict stirs up a negative image: a person of low moral character who willfully chooses to engage in questionable behavior. This image is perpetuated in the media; on a recent episode of E.R., the chief surgeon criticizes another doctor for allowing a heroin addict (who has been treated for an abscess) to exchange a dirty needle, explaining "we donà ¢t want these low-lives hanging around the hospital." The social stigma attached to addicts reflects the great gap that exists between scientific knowledge and public perception of addiction. Just as mental illness was viewed as a social problem instead of a medical issue until the last several decades, drug addiction continues to be seen as a character flaw instead of as the biological problem that it is. As defined by the American Psychiatric Association, addiction is a "chronically relapsing disorder that is characterized by three major elements: (a) compulsion to seek and take the drug, (b) loss of control in limiting intake, and (c) emergence of a negative emotional state when access to the drug is prevented" (1). This disorder results from the repeated use of a drug over a prolonged period of time, causing physical changes in the brain. Perhaps the most addictive of drugs is cocaine. Cocaine acts on the mesoaccumbens dopamine (DA) pathway of the midbrain, extending from the ventral tegumental area (VTA) to the nucleus accumbens (NAc). (2). This pathway is also known as the reward pathway as it is the area of the brain that is activated when someone has a pleasurable experience such as eating, sex, or receiving praise. (NOTE: The reward pathway was discovered through the technique of... ...se%2fform%2facademic%2fs_genjourn_more.html%25a3AD_FORM%25a4s_genjourn%25a3T1%25a4cocaine%20addiction%25a3S1%25a4Title%25a3S2%25a4AND%25a3T2%25a4Morris%2c%20Kelly%25a3S3%25a4author%25a3S4%25a4AND%25a3T3%25a4%25a3S5%25a4Title%25a3date%25a4AFT%25a3after%25a45%3aYR%25a3frm_rng%25a4%25a3to_rng%25a4%25a3srccat%25a4GENMED%3bALLJNL%25a3source%25a4$srccat%25a3pubtitle%25a4%22The%20Lancet%22%25a3srclf%25a4%25a3srclf_group%25a4%25a3srclf_title%25a4%25a3&wchp=dGLSzS-lSlAl&_md5=9d45ca951ebfccd3b6596b38d0b83ce7 19) Combining Drug Counseling Methods Proves Effective in Treating Cocaine Addiction , Information on drug counseling http://www.nida.nih.gov/NIDA_Notes/NNVol14N5/Combining.html 20) Coping Skills Help Patients Recognize and Resist the Urge to Use Cocaine , Information on therapy for cue-induced relapse http://www.nida.nih.gov/NIDA_Notes/NNVol13N6/Coping.html

Africans in the Berlin Conference

Africans in the Berlin Conference The Berlin Conference of 1884-1885 established most of the borders of contemporary nation-states on the African continents today. The Conference set in motion the â€Å"scramble for Africa. † Out of the 14 African states no African leader was present at the conference to assert claims to territory and to agree to regulations on trading. Five of the states at the conference did not even end up with any colonies but had more of a say than all the African leaders. The boundaries in Africa, drawn during the Berlin Conference, greatly attributed to the political unrest in Africa.During the Berlin Conference all the participants signed a notion that said they could not have colonies unless they have full control of the territory they encompassed. The European powers had to send expeditionary forces to pacify what were called their African â€Å"spheres of influence. † This process was bloody and ugly due to the amount of Africans fighting bac k. These pacifications lasted all the way up to World War 1. These uprisings caused anarchy in some places in Africa. Only Ethiopia successfully kept their freedom militarily. But all across Africa, hundreds of polities large, and small fought to destroy these superimposed boundaries.Pre-existing political entities found themselves arbitrarily or deliberately divided in two, three, or even four parts. The main Hausa territories were split between French Niger and British Nigeria, the Maasai suddenly had to practice nomadic pastoralism across a line between German and British East Africa. Conflicts in Africa could definitely were partly caused by boundaries drawn during the Berlin Conference. Uganda’s numerous civil conflicts are pure evidence. Also the Nigerian Civil War of 1967-1971 can partly be traced back to the Berlin Conference.Conflicts between states also can be traced to the Conference because of the ridiculous political geography they put in place. Examples of these conflicts are Cameroon and Libya’s with Chad. The finger of Namibia that sticks out across the top of Botswana, known as the Caprivi Strip has been a geopolitical hotspot ever since it appeared on the map. The Berlin Conference was a very unjust and unethical. They drew out boundaries for Africa without the African leaders consent and then took over all of the nations. The Berlin Conference was definitely connected to the political unrest in Africa.

Calculating Limiting Reactant of a Chemical Reaction

Chemical reactions rarely occur when exactly the right amount of reactants will react together to form products. One reactant will be used up before another runs out. This reactant is known as the limiting reactant. Strategy This is a strategy to follow when determining which reactant is the limiting reactant.Consider the reaction:2 H2(g) O2(g) → 2 H2O(l)If 20 grams of H2 gas is reacted with 96 grams of O2 gas, Which reactant is the limiting reactant?How much of the excess reactant remains?How much H2O is produced? To determine which reactant is the limiting reactant, first determine how much product would be formed by each reactant if all the reactant was consumed. The reactant that forms the least amount of product will be the limiting reactant. Calculate the yield of each reactant. The mole ratios between each reactant and the product are needed to complete the calculation:The mole ratio between H2 and H2O is 1 mol H2/1 mol H2OThe mole ratio between O2 and H2O is 1 mol O2/2 mol H2OThe molar masses of each reactant and product are also needed:molar mass of H2 2 gramsmolar mass of O2 32 gramsmolar mass of H2O 18 gramsHow much H2O is formed from 20 grams H2?grams H2O 20 grams H2 x (1 mol H2/2 g H2) x (1 mol H2O/1 mol H2) x (18 g H2O/1 mol H2O)All the units except grams H2O cancel out, leavinggrams H2O (20 x 1/2 x 1 x 18) grams H2Ograms H2O 180 grams H2OHow much H2O is formed from 96 grams O2?grams H2O 20 grams H2 x (1 mol O2/32 g O2) x (2 mol H2O/1 mol O2) x (18 g H2O/1 mol H2O)grams H2O (96 x 1/32 x 2 x 18) grams H2Ograms H2O 108 grams O2O Much more water is formed from 20 grams of H2 than 96 grams of O2. Oxygen is the limiting reactant. After 108 grams of H2O forms, the reaction stops. To determine the amount of excess H2 remaining, calculate how much H2 is needed to produce 108 grams of H2O.grams H2 108 grams H2O x (1 mol H2O/18 grams H2O) x (1 mol H2/1 mol H2O) x (2 grams H2/1 mol H2)All the units except grams H2 cancel out, leavinggrams H2 (108 x 1/18 x 1 x 2) grams H2grams H2 (108 x 1/18 x 1 x 2) grams H2grams H2 12 grams H2It takes 12 grams of H2 to complete the reaction. The amount remaining isgrams remaining total grams - grams usedgrams remaining 20 grams - 12 gramsgrams remaining 8 gramsThere will be 8 grams of excess H2 gas at the end of the reaction.There is enough information to answer the question.The limiting reactant was O2.There will be 8 grams H2 remaining.There will be 108 grams H2O formed by the reaction. Finding the limiting reactant is a relatively simple exercise. Calculate the yield of each reactant as if it were completely consumed. The reactant that produces the least amount of product limit the reaction. More For more examples, check out Limiting Reactant Example Problem and Aqueous Solution Chemical Reaction Problem. Test your new skills by answering  Theoretical Yield and Limiting Reaction Test Questions. Sources Vogel, A. I.; Tatchell, A. R.; Furnis, B. S.; Hannaford, A. J.; Smith, P. W. G. Vogels Textbook of Practical Organic Chemistry, 5th Edition. Pearson, 1996, Essex, U.K.Whitten, K.W., Gailey, K.D. and Davis, R.E. General Chemistry, 4th Edition. Saunders College Publishing, 1992, Philadelphia.Zumdahl, Steven S. Chemical Principles, 4th Edition. Houghton Mifflin Company, 2005, New York.

Working The Angles The Shape Of Pastoral Integrity

In today s culture the congregational expectations on a pastor are quite different from the vocational spiritual disciplines required for faithful pastoral ministry. The office of the pastor, for the congregation, has become nothing more than a managerial position of running a religious organization. In response to this Eugene Peterson offers his take on the distinctive work of the pastor and the practices that shape pastoral integrity. In his book, Working the Angles: The Shape of Pastoral Integrity, Peterson outlines the practices of prayer, Scripture and spiritual direction as the backbone of faithful pastoral ministry. The following will discuss these practice, there benefit to pastoral ministry and why Peterson call to these practice are important to pastors today. Peterson opens Working the Angles with three chapters on prayer. He critiques contemporary Seminary culture for training pastors to focus on the ministry of the Word and Sacrament. However, Peterson contests, â€Å"F or the majority of the Christian centuries most pastors have been convinced that prayer is the central and essential act for maintaining the essential shape of the ministry to which they were ordained.†1 For Peterson prayer is never the first word, but it is response to what God has already done and is doing. Therefore, the pastoral task is to restore prayer to this position of response through practice of praying the Psalms and other Scripture. It is vitally important that pastors make space forShow MoreRelatedMetz Film Language a Semiotics of the Cinema PDF100902 Words   |  316 Pagessignifier (or expression) and the significate (or content) of the linguistic unit, but the second articulation can affect only the signifier of its corresponding unit. Monemes may be lexical or grammatical. The sentence We are working contains four monemes; the word working is constituted of the lexical moneme (or semanteme, or, as Mar* De Saussure s translator used the terms signifier/signified. The translator of Andrà © Martinet s Elements de linguistique gà ©nà ©rale prefers significans/significatumRead MoreProject Managment Case Studies214937 Words   |  860 PagesManager s Day (B) (see handout provided by instructor) An International Project Manager s Day (C) (see handout provided by instructor) An International Project Manager s Day (D) (see handout provided by instructor) Ellen Moore (A): Living and Working in Korea 177 Ji nan Broadcasting Corporation 196 4 PROJECT MANAGEMENT ORGANIZATIONAL STRUCTURES 205 Quasar Communications, Inc. 207 Jones and Shephard Accountants, Inc. 212 Fargo Foods 216 Government Project Management 220 Falls EngineeringRead MoreFundamentals of Hrm263904 Words   |  1056 Pagesat Ethics 22 Summary 23 Demonstrating Comprehension: Questions for Review 24 Key Terms 24 HRM Workshop 25 Linking Concepts to Practice: Discussion Questions 25 Developing Diagnostic and Analytical Skills 25 Case 1: Work/Life Balance at Baxter 25 Working with a Team: Understanding Diversity Issues 25 Learning an HRM Skill: Guidelines for Acting Ethically 26 Enhancing Your Communication Skills 26 ETHICAL ISSUES IN HRM: Invasion of Privacy? 9 WORKPLACE ISSUES: We Are Now Entering the Blogosphere

Essay about Communication - 1427 Words

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Content Audit or Inventory Public Money

Question: Describe about the Content Audit or Inventory for Public Money. Answer: Introduction In large listed companies, public money is involved and so it is necessary that the financial information presented to the stakeholders is free from any material misstatements. An audit serves as a check on the compliance and includes areas like quality management and secretarial due diligence. The audit provides a fair opportunity for the evaluation of the risk management policies and also the effectiveness of the control and governance procedures. For all these reasons, an audit would be beneficial to the entity in more ways than one and hence it is considered to be indispensible. Involvement of the Audit The Audit involves an examination of the books of accounts, vouchers, documents and all such statutory records of the company by an independent auditor. This process helps in ascertaining the true and fair nature of the financial statements. The flaws in the internal controls, processes or the accounts are pointed out and the required measures for the proper maintenance as required by the law are explained by the auditors and the appropriate measures can be taken for the same. The Auditor performs the examination based on the substantive procedures and obtains evidences and evaluates the same based on professional judgment and the opinion of the auditor is communicated to the users through the audit report (Messier Emby, 2005). An auditor first obtains the understanding of the business, identifies the areas that could be potentially materially incorrect, verification of the accuracy of the account balances and transactions and appropriate disclosures, testing the effectiveness of the internal controls and deciding whether the accounting policies are reasonable, assessment of the reasonableness of the estimates and judgments based on which the financial statements are prepared by the management, evaluation of the significant events that have occurred after the balance sheet date and qualifying the audit report for any such material inconsistencies or disagreement with the management or violation of law (Kaplan, 2011). Auditor With reference to a company, the audit refers to the financial audit in which the auditor is required to report the users of the financial statements about the adequacy of the financial reporting and the statutory book keeping. The company can appoint any of the below who are having a valid registration with ASIC and qualified for auditor appointment: An individual (registered for company audit) or A firm or A company (authorized Audit Company). For becoming a registered auditor, years of professional studies and training is required. Even after becoming a registered auditor, the Continuous Professional Training and learning is a mandatory requirement. As auditors are synonymous with trust and integrity, misconduct and violation of principles becomes an offence and such auditors are disqualified and banned by ASIC to perform any audits. A list of such banned auditors is available on the ASIC website. The appointment of auditor is usually done by the shareholders in a general meeting but if they fail to do so, then the directors may appoint the auditor. The first auditor of the company is usually appointed by the Board of Directors within one month from the registration of the company who holds office until a general meeting of the company in which either a new auditor is appointed by the members or the existing auditors appointment or term is confirmed (Parker et. al, 2011). An Auditor holds office until he is removed by the company as per S.329 or the auditor is deceased or the auditor obtains the consent of ASIC to resign as per S.342 or the auditor has to retire from office according to a few provisions covered by S.327 or if the company is winding up or the auditor suffers incapability due to the Corporations Act, Division 2 Part 2M.4. Third party (the auditor) access to their confidential business information Auditor is covered by the statutory responsibility to carry out a verification and examination of the books of accounts and financial statements. An auditor can obtain all such information that is necessary for conducting the audit. The basic qualities that every auditor is expected to possess are integrity, objectivity, confidentiality, knowledgeable and independence. According to these guidelines, where the auditor has obtained knowledge of business during audit, such knowledge should not be used by the auditor in setting up a business for his own benefit or shared with relatives of the auditor or with the entitys competitors. In such way, confidentiality is a necessary requirement due to which the profession of auditors is looked upon with respect. Hence, a good auditor does not indulge in such practices and so it is completely safe to entrust the auditor books of accounts and all other such information required for carrying out the audit. Non-audit of the accounts With the expansion in international trade, business structures have become complex and dynamic; there might be various laws and regulations subject to the business and a proper understanding and compliance of the same is required to prevent big sums of penalties. The innumerable amendments and updating of the legal provisions might anytime trigger the requirements of lodging returns or such other compliances without the Corporate realizing it. There are even companies that run business for years without the knowledge of legal provisions and then suffer a costly compliance exercise (Heeler, 2009). When it comes to a sole trader business or even a small partnership set up, audit might not be mandatorily required. But once a company is formed, it is governed by the ASIC on one side with reference to the taxation matters and the Corporations Act on the other side for all the compliance related matters. As the Board of Directors is busy with day to day affairs of running the business profitably, accounts are often overlooked. Hence, audit is required for Businesses to find out how they are performing. The Shareholders or members of the company have entrusted the running of the business on the Directors. If accounts are not audited, the shareholders might point out to the Directors for any fraud or mismanagement or financial irregularities (Gilbert et. al, 2005). If audit is carried out, the management is given a report of the potential lapse in the internal controls along with the recommendations for rectifying the same thus reducing the risk intensiveness of the entity (Gilbert et. al, 2005). Overall the productivity, efficiency, cost effectiveness, proper and systematic tax planning can all be done by conducting an audit. When the company tries to obtain a bank loan, or is having talks for mergers, acquisitions, joint ventures or such other proposals, then audited financial statements are valued and also lend credibility. If audit is not done, there are chances to miss out on a profitable venture. Thus if an audit is not carried out, then all these benefits will be zeroed down. Audit by Jane Jane is holding 40% shareholding in the company and is also in charge of the day to day management of the company. Even as per the code of ethics, a partner or relative of any of the directors cannot perform the audit to ensure the quality of independence is maintained (Baldwin, 2010). As Jane is a shareholder and also in charge of the management, it is not possible that Janes mistakes are discovered by Jane herself. For this reason also, Jane cannot conduct the audit herself. Even if Jane is a qualified auditor, she cannot perform the audit for the above stated reasons. Using of any information beforehand by any officials is a serious offence. In case the auditors found misusing the business information in any way, for financial or non financial interest, then the client company or the entity can sue the auditor for professional misconduct. This might even lead to ASIC disqualifying or banning the auditor which brings disrepute to the auditor. Confidentiality and independence are considered to be the key traits of an auditor without which the auditor cannot become successful. Therefore, considering such dispute it is essential that the company should have different auditor so that any problem is created. Audit by firm For conducting the audit, one of the crucial factors is independence. This quality requires that the auditor conducts the audit without being influenced by any financial or any other factors that could lead to a biased opinion. Thus if an independent party is appointed as the auditor, who is neither a relative of any of the shareholders or directors or materially interested in the company, then such audit would provide an effective opinion as to how an outsider would look at and analyze the business (Cappelleto, 2010). Choosing a particular firm will create plenty of advantages for the business because it lead to immense transparency and hence, will create a strong level of trust. There could also be cases where the business is currently running profitably and economically, but the auditor has seen cases of potential default in some other clients and companies and so the awareness about the same can be passed on to the existing client and sufficient due diligence exercise can be carr ied out to prevent such defaults due to which other companies have suffered. Audit by firm carries a great deal of advantage because proper information is provided and hence, it intimates the related parties about any adverse situation. Moreover, it enhances the goodwill of the company. Audit by firm leads to better understanding and a level of professional accuracy is maintained because in the case of any fraud or negligence, the firm will be held liable and responsible. Therefore, an audit by firm helps the business to prosper. Conclusion From the above report, it is clear that the selection of auditor must be done with precision because the entire process of reporting depends on it. The involvement of the auditor and providing an independent decision is important therefore, before selection of auditor it must be kept in mind that the auditor should not be related. From the above report, it is clear that Jane is a shareholder and hence cannot be an auditor because of the vested interest. It will impact the quality of the decision. 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