Ebola Virus Mechanism of Infection

Ebola Virus Mechanism of Infection The Ebola virus (EBOV) is an enveloped, non-segmented, negative-strand RNA virus, which  together with Marburg virus, makes up the filoviridae family. The virus causes severe  hemorrhagic fever associated with 50-90% human mortality1. Four species of the virus (Zaire,  Sudan, Cà ´te d’Ivoire, and Reston ebolavirus) have thus far been identified, with Zaire typically  associated with the highest human lethality2. A fifth EBOV species is confirmed in a 2007  outbreak in Bundibugyo, Uganda3,4. Infection with EBOV results in uncontrolled viral  replication and multiple organ failure with death occurring 6-9 days after onset of  symptoms5. Fatal cases are associated with high viremia and defective immune responses,  while survival is associated with early and vigorous humoral and cellular immune  responses6-9. Although preliminary vaccine trials in primates have been highly  successful10-13, no vaccines, specific immunotherapeutics, or post-exposure treatments are  currently approved for human use. Since 1994, EBOV outbreaks have increased more than  four-fold, thus necessitating the urgent development of vaccines and therapeutics for use in the  event of an intentional, accidental or natural EBOV release. The EBOV genome contains seven genes, which direct the synthesis of eight proteins. Transcriptional editing of the fourth gene (GP) results in expression of a 676-residue transmembrane-linked glycoprotein termed GP, as well as a 364-residue secreted glycoprotein  termed sGP14,15. EBOV GP is the main target for the design of vaccines and entry inhibitors. GP is post-translationally cleaved by furin16 to yield disulfide-linked GP1 and GP2  subunits17. GP1 effects attachment to host cells, while GP2 mediates fusion of viral and host  membranes16,18-20. EBOV is thought to enter host cells through receptor-mediated  endocytosis via clathrin-coated pits and caveolae21, followed by actin and microtubuledependent  transport to the endosome21, where GP is further processed by endosomal  cathepsins22-24. Essential cellular receptor(s) have not yet been identified, but DC-SIGN/LSIGN25,  hMGL26, ÃŽ ²-integrins27, folate receptor-ÃŽ ±28 and Tyro3 family receptors29 have all  been implicated as cellular factors in entry. Here, we report the crystal structure of EBOV GP,  at 3.4 Ã… resolution, in its trimeric, pre-fusion conformation in complex with neutralizing  antibody Fab KZ52. GP1 is responsible for cell surface attachment, which is probably mediated by a region  including residues 54-20132. GP1 is composed of a single d omain (∠¼65 Ã… Ãâ€" 30 Ã… Ãâ€" 30 Ã…),arranged in the topology shown in Fig. 2a, and can be further subdivided into the (I) base, (II)  head and (III) glycan cap regions (Fig. 2a and Supplemental Fig. S3). The base (I) subdomain  is composed of two sets of ÃŽ ² sheets, forming a semi-circular surface which clamps the internal  fusion loop and a helix of GP2 through hydrophobic interactions (Fig. 2b). Moreover, this  subdomain contains Cys53, which is proposed to form an intermolecular disulfide bridge to  Cys609 of the GP2 subunit17. Cys53 resides near GP2 in the ÃŽ ²2-ÃŽ ²3 loop at the viral membraneproximal  end of the base subdomain (Fig. 2a-b). Our EBOV GP contains an intact GP1-GP2  disulfide bridge, based on reducing and non-reducing SDS-PAGE analysis. However, the region containing the counterpart GP2 cysteine is disordered, which may reflect functionally  important mobility in the region. The head (II) is located between the base and glycan cap   regions towards the host membrane surface. Two intramolecular disulfide bonds stabilize the  head subdomain and confirm the biochemically determined disulfide bridge assignments17.   Cys108-Cys135 connects a surface-exposed loop (ÃŽ ²8-ÃŽ ²9 loop) to strand ÃŽ ²7, while Cys121-Cys147 bridges the ÃŽ ²8-ÃŽ ²9 and ÃŽ ²9-ÃŽ ²10 loops (Fig. 2a). The glycan cap (III) contains four  predicted N-linked glycans (at N228, N238, N257 and N268) in an ÃŽ ±/ÃŽ ² dome over the GP1  head subdomain (Fig. 1b and 2a). This subdomain does not form any monomer-monomer  contacts and is fully exposed on the upper and outer surface of the chalice. The central ÃŽ ² sheets  from the head and glycan cap together form a fairly flat surface and, in the context of the GP  trimer, form the three inner sides of the chalice bowl. Ebola virus GP2 GP2 is responsible for fusion of viral and host cell membranes and contains the internal fusion  loop and the heptad repeat regions, HR1 and HR2. Many viral glycoproteins have fusion  peptides, located at the N terminus of their fusion subunit, which are released upon cleavage  of the precursor glycoprotein. By contrast, class II and class III fusion proteins, as well as class  I glycoproteins from Ebola, Marburg, Lassa and avian sarcoma leukosis viruses, contain  internal fusion loops lacking a free N terminus. The crystal structure reveals that the EBOV  GP internal fusion loop, which encompasses residues 511-556, utilizes an antiparallel ÃŽ ²Ã‚  stranded scaffold to display a partially helical hydrophobic fusion peptide (L529, W531, I532,  P533, Y534 and F535) (Fig. 2c). The side chains of these hydrophobic residues pack into a  region on the GP1 head of a neighboring subunit in the trimer, reminiscent of the fusion peptide  packing in the pre-fusion parainfluenz a virus 5 F structure33. A disulfide bond between Cys511  at the base of ÃŽ ²19 and Cys556 in the HR1 helix covalently links the antiparallel ÃŽ ² sheet. This  disulfide bond between the internal fusion loop and HR1 is conserved among all filoviruses,  and is analogous to a pair of critical cysteines flanking the internal fusion loop in avian sarcoma  leukosis virus34,35. Interestingly, the EBOV internal fusion loop has features more similar to  those observed in class II and III viral glycoproteins (in particular to flaviviruses) than those  previously observed for class I glycoproteins (Supplemental Fig. S4). It thus appears that  regardless of viral protein class, internal fusion loops share a common architecture for their  fusion function. EBOV GP2 contains two heptad repeat regions (HR1 and HR2), connected by a 25-residue  linker containing a CX6CC motif and the internal fusion loop. The crystal structures of postfusion  GP2 fragments30,31 have revealed that the two heptad repeat regions form antiparallel  ÃŽ ± helices and that a CX6CC motif forms an intrasubunit disulfide bond between Cys601 and  Cys608 (Supplemental Fig. S5). In the pre-fusion EBOV GP, HR2 and the CX6CC motif are  disordered. By contrast, the HR1 region is well ordered and can be divided into four segments:  HR1A, HR1B, HR1C and HR1D (Fig. 2c), which together assemble the cradle encircling GP1. Similarly, heptad repeat regions in influenza and parainfluenza viruses also contain multiple  segments in their pre-fusion helices that substantially rearrange in their post-fusion  conformations33,36,37. The first two segments, HR1A and HR1B (residues 554-575), together form an ÃŽ ± helix with an  Ã¢Ë† ¼40 ° kink at T565, which delineates HR1A from HR1B. Interestingly, the bend between  HR1A and HR1B contains an unusual 3-4-4-3 stutter, which may act as a conformational  switch31, rather than the typical 3-4 periodicity of heptad repeats (Supplemental Fig. S6). A  similar stutter has also been noted in parainfluenza virus 5 F33. The Ebola virus HR1C (residues  576-582) forms an extended coil linking HR1B to the 14-residue ÃŽ ± helix of HR1D (residues  583-598). HR1D forms an amphipathic helix and the hydrophobic faces of each HR1D join to  form a three-helix bundle at the trimer interface. Although the breakpoint maps directly to a  Lee et al. Page 3 Nature. Author manuscript; available in PMC 2009 June 22. NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript  chloride ion binding site in the post-fusion conformation of GP230,31 and at least two other  viruses38,39, no chloride ion is observed here as HR1 and HR2 do not come together to form  the six-helix bundle. Instead, the pre-fusion GP2 adopts a novel conformation, intimately  curled around GP1 (Fig. 1c). Ebola virus GP-KZ52 interface KZ52 is an antibody isolated from a human survivor of a 1995 outbreak in Kikwit, Democratic  Republic of the Congo (formerly Zaire)40. This antibody neutralizes Zaire ebolavirus in  vitro40 and offers protection from lethal EBOV challenge in rodent models41, but has minimal  effects on viral pathogenicity in non-human primates42. KZ52 is directed towards a vulnerable,  non-glycosylated epitope at the base of the GP chalice, where it engages three discontinuous  segments of EBOV GP: residues 42-43 at the N terminus of GP1, and 505-514 and 549-556  at the N terminus of GP2 (Fig. 3 and Supplemental Fig. S7). Although the majority of the GP  surface buried by KZ52 belongs to GP2, the presence of both GP1 and GP2 are critical for  KZ52 recognition43. It is likely that GP1 is required to maintain the proper pre-fusion  conformation of GP2 for KZ52 binding. Indeed, KZ52 is the only antibody known to bridge  both attachment (GP1) and fusion (GP2) subunits of any viral gly coprotein. Given that KZ52  requires a conformational epitope seen only in the GP2 pre-fusion conformation and that the  KZ52 epitope is distant from the putative receptor-binding site (RBS), KZ52 likely neutralizes  by preventing rearrangement of the GP2 HR1A/HR1B segments and blocking host membrane  insertion of the internal fusion loop. Alternatively, IgG KZ52 may sterically hinder access to  the RBS or to a separate binding site of another cellular factor, especially if multiple attachment  events are required for entry. The KZ52 epitope of GP is convex and does not have a high shape complementarity to the  antibody (Sc index of 0.63), although ∠¼1600 Ã…2 of each GP monomer are occluded upon KZ52  binding. The antibody contacts a total of 15 GP residues by van der Waals interactions and 8  direct hydrogen bonds (Supplemental Fig. S7). Ten out of 15 residues in the structurally defined  KZ52 epitope are unique to Zaire ebolavirus (Supplemental Fig. S6), thus explaining the Zaire  specificity of KZ52. Ebola virus GP glycosylation We generated a fully glycosylated molecular model of EBOV GP to illustrate the native GP  trimer as it exists on the viral surface (Fig. 4). The majority of N-linked glycosylation sites are  concentrated in the mucin-like domain and glycan cap of GP1. Given that the mucin-like  domain is ∠¼75 kDa in mass (protein and oligosaccharide), the volume of this domain is  predicted to be similar to each GP monomer observed here. The crystal structure suggests that  the mucin-like domain is linked to the side of each monomer and may further build up the walls  of the chalice, forming a deeper bowl (Fig. 4). Although a mixture of complex, oligomannose  and hybrid-type glycans are found on intact, mucin-containing GP144, those glycans outside  the mucin-like domain are likely to be complex in nature: the mucin-deleted GP used for  crystallization is sensitive to PNGaseF, but not to EndoH treatment (Supplemental Fig. S8).   Modeling of complex-type oligosaccharides on the EBOV GP indicates that the majority of  the GP trimer is cloaked by a thick layer of oligosaccharide, even without the mucin-like  domain (Fig. 4). The ∠¼19 additional oligosaccharides on the full-length GP (17 on the mucinlike  domain and 2 more on GP1, disordered here) further conceal the sides and top of the  chalice. The KZ52 binding site and, presumably, the flexible regions of HR2 and the  membrane-proximal external region (MPER) remain exposed and perhaps vulnerable to  binding of antibodies and inhibitors. Lee The development of neutralizing antibodies is limited in natural Ebola virus infection. Many  survivors have low or insignificant titres1,7, and those antibodies that are elicited preferentially  recognize a secreted version of the viral glycoprotein that features an alternate quaternary  structure and lacks the mucin-like domain43. The glycocalyx surrounding EBOV GP likely  forms a shield that protects it from humoral immune responses and/or confers stability insideor outside a host. The mucin-like domain and glycan cap sit together as an external domain to  the viral attachment and fusion subunits, reminiscent of the glycan shields of HIV-1  gp12045,46   and Epstein-Barr virus gp35047, perhaps pointing to a common theme for immune  evasion. Alignment of filoviral sequences indicate that regions involved in immune evasion  have a low degree of sequence conservation [i.e. GP1 glycan cap (∠¼5%) and mucin-like domain  (0%)], but the N-glycosylation sites in the glycan cap are mostly conserved among all EBOV  subtypes (Supplemental Fig. S6), indicating the functional importance of these posttranslational  modifications. Sites of receptor binding and cathepsin cleavage Although a definitive receptor for EBOV remains to be identified, previous studies32,48,49  have determined that residues 54-201, which map to the base and head subdomains of GP1,  form a putative receptor-binding site (RBS) for attachment to host cells. Additional  experimental studies have identified at least 19 GP1 residues, assigned into four groups based  on the location in the structure, that are critical for viral entry48-50 (Fig. 5). Many of these  residues are apolar or aromatic and are involved in maintaining the structural integrity of GP1  for receptor binding or fusion. However, six residues (K114, K115, K140, G143, P146 and  C147) cluster within a ∠¼20 Ãâ€" 15 Ã… surface in the inner bowl of the chalice and may thus  represent important receptor contact sites. All residues in the putative RBS are highly conserved  among Ebola virus species (Supplemental Fig. S6). Importantly, this putative RBS is recessed beneath the glycan cap and perhaps further masked  by the mucin-like domain (Fig. 4), suggesting that additional conformational change or removalof the mucin-like domain could reveal additional surfaces required for receptor or cofactor  binding. It has been demonstrated that endosomal proteolysis of EBOV GP by cathepsin L  and/or B removes the mucin-like domain to produce a stable ∠¼18 kDa GP1 intermediate which  has enhanced viral binding and infectivity22-24. The precise site of cathepsin cleavage is  unknown and the role of cathepsins in natural infection is as yet unclear. However, formation  of an ∠¼18 kDa GP1 fragment implies that cathepsin may cleave near the GP1 ÃŽ ²13-ÃŽ ²14 loop  (residues 190-213). Indeed, this loop is unresolved in the pre-fusion structure, suggesting  enhanced mobility and accessibility to enzymatic cleavage. Cleavage within this loop would  remove the entire mucin-like domain and glyc an cap region (Fig. 5). As a result, ÃŽ ²7 to ÃŽ ²9  strands and their associated loops would become exposed. These regions of GP are in proximity  to the previously identified residues critical for viral entry. The fold, location and  physicochemical properties of this site should now provide new leads in the search for the  elusive filoviral receptor(s). A summary of the Ebola virus mechanism of infection, including the events of cathepsin  cleavage and conformational changes to GP2 during fusion, is presented.

Health and Social

Hollie Kelly- D1 Justify how the two-day diet plan meets the dietary needs of the two service uses Type-two diabetes Roger I have transformed Rogers two day diet plan to ensure he does not take in low carbohydrates as they are high in fats. I have only used complex carbohydrates to form part of Roger’s diet. I have tried to keep Rogers diet low in fat and sugar with only obtaining a small amount of fat and sugar in his two-day diet plan. In general, people with type 2 diabetes have a lifespan that is five to ten years less than those without the disease.The most common long-term effect of type 2 diabetes is damage to blood vessels. Because of this, diabetics are twice as likely to develop cardiovascular disease, which can result in blocked arteries, and eventually lead to a stroke or heart attack. The main cause of death in type 2 diabetes sufferers is cardiovascular disease and associated complications. Obese Susan I have transformed Susan’s two-day diet plan to ensure her calorie intake isn’t as high as it was. I have maintained a balanced diet that should fill her up for both days. I have ensured her intake of fats and sugar is to a minimum.I have balanced out her intake of dairy products, carbohydrates and protein so it helps her lose weight faster to obtain a healthy lifestyle. Obesity can reduce your life expectancy by up to 9 years and many chronic diseases can be prevented by maintaining a healthy weight. Being overweight can also put extra pressure on joints and limbs, making activity quite difficult and sometimes any movement at all can be painful. Other physical problems caused by obesity are that obese women who become pregnant have a higher risk pregnancy that than of a healthy weight.

Management Accounting Case Study - 2102 Words

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Prevention And Prevention Of Vaccination - 1417 Words

Imagine a world ravaged by disease and death. It started with one person in a remote village in Africa or Asia. Slowly the disease spread throughout the village infecting hundreds. Soon the outbreak jumps to neighboring villages and eventually towns. Thousands begin to become infected and death is not far away. As the disease starts to spread from town to city to country to continent millions begin to die. The disease is tearing through the globe causing an epidemic and wiping millions out. The disease seemed something minor that did not need a vaccination when health officials first warned of it. First world countries were bullied into believing that their children were at risk for neurological damage that had no scientific data behind it because it seemed scary. Parents stopped having their children receive vaccines making them susceptible to the disease and its wrath. This an extreme example of a potentially real scenario, but luckily there is a way to prevent something like this from becoming reality: vaccination. According to the United States of America Centers for Disease Control and Prevention, or the CDC, a vaccine is inputting a specific disease, such as Chickenpox, into the body imitating the symptoms of the real disease or infection while not actually giving the receiver the symptoms in most cases. The body then takes action by creating white blood cells as a defense which attacks and kills the infection. This then leaves extra cells that the body stores in aShow MoreRelatedPrevention And Prevention Of Vaccination1405 Words   |  6 Pagesvaccines, making them susceptible to the disease and its wrath. This an extreme example of a potentially real scenario, but luckily there is a way to prevent something like this from becoming reality: vaccination. 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Americ Are We Really Free - 903 Words

America: Are we really free? Every day, nearly one-third of U.S. children ages four to nineteen eat from a fast food restaurant this adds on about six extra pounds per child every year and increases their risk of obesity (Fast Food). In my research I learned a lot about the health risks that come with eating fast food. Fast food causes childhood obesity rates in America to increase because of its convenience, ingredients, and lack of telling people what is in the food. The convenience of fast food plays a major role in how often people eat it. When driving down a main road chances are you will see a countless number of fast food restaurants and drive thrus. I know that it saves time and money for people who are coming home from work but people should plan out their meals nights before. One generation ago three quarters of the money that is used to buy food was spent on fresh food to cook at home. Now almost half of that same money is used to purchase fast food (Introduction). Fast food is also cheap which adds to the reasons why people enjoy it. The parents will allow the children to order what they want because chances are it only will cost them a couple of dollars. Childhood obesity rates go up as the prices of fast food goes down (Introduction). In 1970 over six billion dollars were spent on fast food. In 2014 one hundred ninety-five dollars were spent (Kelly). The fast food industry is aware of the power it has over American citizens and it is going to use that powerShow MoreRelatedHistorical Anyalysis Essay2125 Words   |  9 PagesHistory is formed from analyzing evidence from the past and making a connection. Many historians have different personal perspectives on history, and by using the historical method they can all draw conclusions as to what happened and advocate a theory. We find that history is much more than what happened in the past from reading, The Strange Death of Silas Deane. This is a good example of applying the historical methods and producing a new theory based on reliable evidence. The author classified theRead More African Minkisi and American Culture Essay6248 Words   |  25 Pagescall moquisie [minkisi] everything in which resides, in their opinion, a secret and incomprehensible virtue to do them good or ill, and to reveal event of past and future† (Williams, 13). The term illness, in this context, is quite different than what we refer to illness. Illness, to the BaKongo, meant anything f rom sickness, to loss of property, and the inability to succeed in things like school and work. . â€Å"The perpetual struggle with the unseen forces that cause illness and misfortunes was (and is)

Political-Legal Environment Affecting the Economy of an Industry Free Essays

Political- legal environment is one of the economic environment that its development has strongly impacts on the marketing decisions of the company. The political environment consists of laws, government agencies, and pressure groups that influence or limit various organizations and individuals in a given society. Each factors of political environment has its own functions and impacts on the economic activities of the industry. We will write a custom essay sample on Political-Legal Environment Affecting the Economy of an Industry or any similar topic only for you Order Now The economic systems work best with at least some regulation. When an industry with well-conceived regulation, it can have more opportunities in competition and ensure fair market for goods and services. The public policy is a set of laws and regulations that the government makes to guide commercial or to limit business for the good of society as a whole. Almost marketing activites requires a wide range of laws and regulations to stable the system and help to make true marketing decisions. Legislation affect business around the world and it has increased steadily over the years such as local-content laws, import restrictions, tax controls, price controls, labor problems. .etc†¦China is one of the Samsung market and it has many laws relates to some issues such as custom import and trading, foreign investment, information technology, pricing and etc†¦. o regulate the activites or the development of Samsung electronics industry in China, for example: Provisional Regulations on Trade Administration of Technology Introduction and Equipment Importation 1- 1996, Implementation Rules on Small-Volume Frontier Trade of Importing Machinery and Electronic Products, Notice on Policies for Reform and Adjustment of Import Tax  , Regulation on Administration of Organizations in Charge of Import or Machinery and Electronic Products in Localities and Departments at the Central Level†¦etc†¦ and through those laws, Samsung enterprises located in China must comply wit h all the articles of Chinese laws and all the regulations that they sign together. The regulations are constantly changing and the marketer must work hard to keep up with changes in regulations and their interpretations.. . . Another example is the activites of Samsung electronics VietNam (SEV) in export and paying tax. According to some regulations and laws on taxes, the contributions to the budget through taxes including import or export taxes, value added tax (VAT) and environmental protection taxes. The political sanctions, which protect the companies from each other. Although business executives may praise competition, they sometimes try to neutralize it when it threatens them, so laws are passed to defined and prevent unfair competition. For example, Korean companies have been increasingly subject to the extraterritorial reach of foreign antitrust laws, in 2005, the US department of Justice(DOJ) imposed fines of $300 million on Samsung electronic, a Korean manufactuer of dynamic random access memory(DRAM) and its US subsidiary, Samsung semiconductor, for participating in an international conspiracy to fix prices in the DRAM market. In addition, according to local reports, the Japan fair trade commission will fine affiliates of Samsung electronics an unprecented amount of several billion yen for price fixing. This was because their sale to subsidiaries of Japanese companies indirectly affected the Japanese market. The Japanese sanctions also protect the right of their authorities and consumers. Another example is the patent infringement between Samsung and Apple. According to US infringement law, US Court ordered  Samsung  to pay $1. 05 billion damages, after ruling that the South Korean firm infringed on some  Apple  patents in  patent infringement trial between Samsung and Apple. And after that, Samsung’s entire line of galaxy were banned in Europe such as the Galaxy S II the Galaxy Tab 7. 7†¦ The government regulation also help to protect the interests of society against unrestrained business behavior. For example, Samsung had been accused by  China Labor Watch  of countenancing illegal and even inhumane conditions at its factories in China, and the group noted that the workers’ overtime at the facilities could reach or often times exceed 100 hours a month. Under current Chinese law, overtime is limited to 36 hours a month. After that, Samsung quickly moved to address the problem also further suggests that Samsung is taking this very seriously. How to cite Political-Legal Environment Affecting the Economy of an Industry, Papers

A Case Study Analysis On Managing Business Operations of ASDA @UK

Question: With reference to the organisation that you studied for ASDA UK and the key leadership and management issues identified affecting its performance, you are now required to recommend how the organisation could resolve these issues. Answer: Executive summary The purpose of the study is to analyze the major issues related with ASDA Company. A different issue has been identified but issue relates with employees motivation and satisfaction has been majorly focused. The problem has been discussed briefly and different motivation theory that is Herzberg Theory of Motivation and Maslow Hierarchy Needs Theory in order to briefly explain and understand the situation. Lastly, recommendations have been provided that can help ASDA to increase employee engagement. Introduction The management of business operations is crucial factor for engaging all the activities of an organization to one point so that set goal can be achieved in due time. The company can fall straight on face if there is gap in the activities and it can lead to management issues that can have long term affect on the company. The purpose of this study is to prepare a report in order to discuss the issues that might be faced by any particular organizations. Therefore, for this study ASDA has been selected which is an UK based company. ASDA is a retail industry or a supermarket chain that is engaged in business of retails food, general merchandise, clothing, financial services and other services. The market share of ASDA is 17.5% and is in second number after Tesco in UK (www.asda.com, 2015). Findings: Key Issues in ASDA Management Issues Employees performance: The issues that is currently associated with ASDA relates with the employees that are not able to enhance their performance to best level. The employees were feeling heavy burden on meeting the daily target due to heavy pressure from the superiors. On the other hand, some employees of the company ASDA were sacked even if the employees take a mere break of seven minute without notifying the supervisor. Therefore, it shows inflexibility in the workplace (Internationalworkplace.com, 2015).Double Charging Issues: The next issue that is faced by the company is complaint of the customers for charging twice by the company. The customer complains that the grocery site of ASDA has a login problem and they cannot login at the time of need (Clear, 2013). Technology Issues in ASDA Online Delivery Problem:The Morley Distribution Centre of ASDA faced a system error that lead to delivery of order in time on the eve of Christmas. Due to the error in the distribution centre almost 200 customers of ASDA suffered delays and around 50 of the customers did not only receive the orders. The technical problem led a bad impression of the supermarket among the customers. The website of the company broke down and due to that customers faced difficulty in receiving and placing orders (MacDonald, 2012). The problem that will be discussed in the brief manner will be related to the management issues of employees motivation. The employees of the company are not able to perform confidently that is lowering down their performance level and personal growth. According to Rayton (2012, pp.1), the employees may not been complete into the task due to lack of proper guidance and support from the leaders or management of the company. The employees performance may dip down if the management or leaders are not taking their responsibility in due manner in order to provide right direction to employees to perform the work in productive manner. Therefore, it can affect the production and supply of quality service to the consumers. On the other hand, Talukdar (2012, pp. 303) mentioned that the inability to perform or adopt effective leadership style can decrease the motivation level of the workers. Therefore, it can decrease the engagement of employee in the work or in management decision making process. In such case, the employee may no longer be interested to remain associated with the ASDA Company and may prefer to resign and work in other companies. Apart from that, Foss and Lindenberg (2012, pp. 372) discussed that before takeover of ASDA by Wal-Mart, the manger of the company effectively linked HR policies with the major policies such as employee and customer satisfaction, quality service, employees can access to top management and had informal working place. Best Employer Award (year) Position 2002 1st 2003 7th 2004 31st 2005 87th After 2006 Was not even in top 100 Discussing employee performance issues with theories Frederick Herzberg Motivational Theory Hygiene Factors Working Conditions : The employees of ASDA may not find the working environment suitable if there is lack of proper equipment and high percentage of dirt in the working place can de-motivate the employees to work in effective manner (Jobmanji.co.uk, 2015). Supervision: The lack of ability and efficiency of the supervisor or leaders to control the behavior of the employees can affect the performance of other workers of ASDA. On the other hand, the inability of the supervision from the management or managers may not be able to provide better direction to employees to complete the work in profitable and efficient manner (Korzynski, 2013, pp. 186). Salary: The discrimination in the declaration of salary can upset the employees. For instance, the women that work in ASDA have issued legal action to the company as they claimed that male workers are paid higher than the women workers in spite of doing jobs of equal level. Therefore, it de-motivated and unsatisfied the employees to work in the company (Internationalworkplace.com, 2015). Company Policy: If the policy of the company is too rigid and harsh then it can decrease the motivation level of the employees. On the other hand, in ASDA the hours of work for the employees is 8 hours in a day but if an employee is asked to work for overtime, the company does not have any firm policy for that and employees are not right paid for giving their extra time. On the other side, the retirement plan policy is only for the higher management not for the low level employees. Therefore, it de-motivates the employees to carry their business activities effectively (Naong, 2012, pp. 487). Motivation Factors Recognition: The Company in order to motivate the employees can provide respect and recognition to them for their work. This would help in making employees more engaged towards work. Responsibility: The employee can be some amount of responsibility to take decision at the time of doing task so that in absence of manager or supervisor they can guide the other new employees to perform the job (Engr.sjsu.edu, 2015). Growth: The employees can be provided better training that can help the workers to have professional and personal growth. The training can be effective in solving the work related issues of employees. The other motivational factors can be work itself, promotion and achievement and other hygiene factors can be relationship with peers or top management, job security and status. Therefore, considering these all factors can help in motivating and encouraging the employees (Uroevi and Miliji, 2012, pp. 247). Maslows Hierarchy of needs Physiological Needs: The management of the company may require providing basic needs of life to workers such as food, cloth, shelter, etc. This would help in motivating the employees to effectively work for the society (Vestal, 2012, pp. 535). Safety needs: The employees may be provided with safety requirement so that they can carry the business activities with ease. The employee laws may be made keeping in view the employees requirement so that they can be protected if any wrong thing happens. Moreover, advanced technology can be used so that workers can be kept safe from any injuries (Huang, 2012, pp. 189). Security needs: The senior management of Asda can provide job security to the efficient employees and can do a life insurance cover for employee that can stimulate and encourage the workers to remain associated with the company. Love and belongingness needs: The manager or supervisor has to provide respect, affection, love, etc. that can help in generating a good feeling in workers. The respect and love can motivate workers to give their best effort and increase productivity. Esteem Needs: According to this need, the workers can be given recognition and respect can be given for their hard work. On the other hand, giving the responsibility can motivate employees to perform and excel well (Talukdar, 2012, 497). Self actualization: If the basic needs and other needs are fulfilled then it can help in developing personal growth among workers and can contribute more towards the organizational goal. Conclusion ASDA is majorly facing problem inside their management in context to employees motivation and satisfaction. The employees are having difficulty in working environment, salary and lack of guidance from the leader that affects the motivation stimuli among the employees. The other issues related with ASDA are online site problem and environmental impact. The issue of employee is great concern for the company because if employees leaves the company than the company would end nowhere. The theory helps in understanding the condition of the case and analyzing the factors that can help to retain and improve employee performance. Recommendations The company has to hire supervisor or leaders that have required skills and efficiency to monitor and direct the employees to increase the performance of workers and generate a feeling of job satisfaction. The quality supervisor should understand the problem that might be faced by the workers and providing solution to them. This would be effective in building relationship with each other. The company has to adopt Training Development Program to enhance the skill of the workers and making them understand the requirement of each job. In training the employees can how to perform the job so that wastage can be done at minimum level. On the other hand, training would help in handling the orders of customers and providing them quality service. In order to solve the double charging issues, the company has to build better and efficient online shopping system so that customers do not face bank charges and customers can shop more reliably. On the other hand, in order to solve the online delivery issue, the company has to hire the expertise people that are capable of handling technical issue and distribution centers so that customer does not have to wait too long for the product. Moreover, the refund policy of the company can be reframed if customer faces problem to technical issues. Reference List Clear, M. (2013). Asda charged twice for my online shopping order: what's Asda doing?. [online] Which? Conversation. Engr.sjsu.edu, (2015). Management Theories, Roles, Motivations and Communication. Foss, N. and Lindenberg, S. (2012). Teams, Team Motivation, and the Theory of the Firm. Manage. Decis. Econ., 33(5-6), pp.369-383. Huang, Q. (2012). Action Research on Motivation in English Reading. TPLS, 2(8). Internationalworkplace.com, (2015). Asda forced to pay out after staff complaint - again. Jobmanji.co.uk, (2015). Reviews From People Who Have Worked At Asda. Korzynski, P. (2013). Employee motivation in new working environment. International Journal of Academic Research, 5(5), pp.184-188. Naong, M. N. (2012). The impact of management indecisiveness on employee motivation and performance. African Journal of Business Management, 6(9). Rayton, B. (2012). "Employee Work Engagement, Affect and Outcomes". Academy of Management Proceedings, 2012(1), pp.1-1. Talukdar, A. (2012). Impact of performance pay on enhanced employee performance: mediating role of work motivation and pay satisfaction. JIBED, 6(3/4), p.303. Uroevi, S. and Miliji, N. (2012). Influence of Demographic Factors on Employee Satisfaction and Motivation. Organizacija, 45(4). Vestal, K. (2012). Which Matters: Employee Satisfaction or Employee Engagement?. Nurse Leader, 10(6), pp.10-11. www.asda.com, (2015). Online Food Shopping, ASDA Direct, George and more at Asda.com. Your.asda.com, (2015). Sustainability Our Approach. Zaptorius, J. (2013). Applying the Priority Distribution Method for Employee Motivation. Business, Management and Education, 11(2), pp.256-280. MacDonald, G. (2012). Asda hit by online Christmas food delivery problems.