Immune Responses of Cytokine Adjuvented DNA Vaccine

Immune Responses of Cytokine Adjuvented DNA Vaccine KOTLA SIVAREDDY Title for the Ph.D Thesis â€Å"Study on Immune responses of Cytokine Adjuvented DNA vaccine (genes coding for structural proteins) for FMD delivered by cationic PLG micro particles†. Objectives: To clone gene coding for structural protein of FMDV in eukaryotic expression vector (pC DNA) under CMV promoter. To clone bovine interleukin 18 gene in suitable cloning vector and express the same in Bacterial/yeast expression system for characterization of expressed recombinant protein. To construct a vaccine construct consisting of FMDV P1 (structural), 3C (Viral protease), and Bovine interleukin 18 genes in eukaryotic expression system under CMV promoter (pC DNA). To study the expression of the vaccine constructs in vitro in BHK-21cells. PLG micro particles’ preparation and characterization. To study the immune responses of the DNA vaccine in experimental animals (guinea pigs / cattle). Foot-and-mouth disease (FMD) caused by foot-and mouth disease virus (FMDV) is an infectious disease affecting cloven-hoofed animals, and poses a serious threat for animal health and exacts an economic toll on the livestock industry. FMD viral genome is a positive-sense single stranded RNA of approximately 8.5kb. The viral RNA genome is translated as a single polypeptide precursor that is subsequently processed by virus-encoded proteases 2A and 3C to produce the structural and non-structural proteins required for virus assembly and replication. One of the initial polypeptide cleavages, mediated by the 2A protein, is a co-translational cleavage at its own C terminus to release it from the 2B protein. The viral 3C proteinase subsequently processes the structural protein precursor, P1-2A, into the capsid proteins, VP0, VP3, and VP1, and the non structural peptide, 2A. These proteins then self assemble to form empty icosahedral capsid particles that contain 60 copies of each protein. Immu nological studies have identified linear and conformational sites that are present on both empty capsids and virions, and antiserum raised against either form has the same serological specificity. Thus, the structural protein precursor, P1-2A, and the 3C protease of FMDV are desirable immune antigens for new vaccine development. In countries where disease eradication has not been achieved, vaccination plays a crucial role in its control. Although inactivated virus vaccines effectively prevent FMD, they have several limitations like short duration of immunity, incomplete viral inactivation and virus escape from vaccine producing facilities. As a result, alternative approaches are being investigated, including the construction of modified live virus, subunit vaccines, synthetic peptides, naked DNA plasmids. DNA vaccination which offers several promising features i.e., DNA is convenient to manufacture and store, its production is safe, sequences from circulating strains can be easily i ncorporated in the vaccines, and it also allows the discrimination of the infected from the vaccinated animals. Several reports have shown the efficiency of DNA vaccination to induce protective immunity in the mouse model. However, the primary difficulty with DNA vaccination is its poor immunogenicity in target species. Cytokines are being used as molecular adjuvants by co administering with DNA vaccines to improve the efficacy of the vaccine. Cytokines play an important role both in the development of a functional immune system as well as in the responses of the organism to infection. Interleukin18 (IL18) is a potent interferon ÃŽ ³ (IFNÃŽ ³) inducing factor (IGIF), enhances Th1 immune responses. Recent studies have shown that IL18 also promote Th-2 type responses and increases dendritic cell (DC) number in lymph nodes in mice. In addition, IL18 has been used as an adjuvant to DNA vaccines for classical swine fever virus, pseudo rabies virus, porcine reproductive and respiratory sy ndrome virus.IL18 was co expressed along with FMDV VP1 in Pichia as fusion protein has enhanced humoral responses and marginally the CMI response in mice. Recombinant fowl pox co-expressing FMDV P1 2A3C and IL18 enhanced the immune responses and gave higher protection in swine Many other studies have shown the positive effect of plasmid encoding the IL-18 as a molecular adjuvant on DNA vaccinations. Efficacy of DNA vaccine could be improved by the inclusion of adjuvants and good vaccine delivery systems. Importantly, cationic microparticle with adsorbed DNA induced enhanced immune responses in comparison to naked DNA and this enhancement was apparent in all species evaluated, including nonhuman primates. Cationic PLG microparticles appear to be effective predominantly as a consequence of the efficient delivery of the adsorbed DNA into DC. Following administration, the micro particles are also very effective at recruiting DC to the injection site, and the micro particles also protect adsorbed DNA against degradation in vivo. A second useful property of micro particles is that they can present multiple copies of antigens on their surface, which has been shown to be optimal for B cell activation. The main advantage of this type of association is the efficient immobilisation of plasmid DNA on the microparticle surface without compromising its integrity. Moreover, after administration, the release of cationic DNA complexes from the surface appeared to facilitate the transfection of cells .At this point, it is not completely understood, whether the adsorption on cationic micro particles can protect plasmid DNA from cleavage through endonucleases after administration in vivo. Nevertheless, release of cationic DNA complexes is expected to provide better protection as compared to release of free DNA. Beside their inherent safety and ease of administration, they improve the DNA capture by antigen presenting cells (APC) and stimulate APC maturation. DNA delivery via PLG has been successfully used to vaccinate against several infections in mice, guinea pigs and even in macaques models. 1. AMPLIFICATION and cloning of gene sequence coding for P12A 3C FMDV serotype ‘Asia1’ gene coding for the polyprotein, P1-2A (2.3 kb) was amplified from viral genome, of the serotype ‘Asia1’ polyprotein gene, using VP4L (Bac) and 2AR (E.coR1) primers. Cloned in to pC DNA at E.co R I, BamHI sites. Ligated and transformed in to DH alpha 5 cells. Transformants are screened by colony PCR by using insert specific primers. Orientation was checked by PCR. Insert release was confirmed by RE digestion by using E.co R I, Bam HI. 3C coding sequences were amplified from clone available in lab. 0.6 Kb was amplified .The purified amplicon was digested and ligated in to p C DNA and transferred in to competent DH5à ¯Ã‚ Ã‚ ¡ cells upon screening by PCR and by re digestion positive clones were conformed. 2. Cytokine amplification (IL18) and cloning in prokaryotic / yeast pC DNA expression vector and characterization of expressed protein. Interleukin 18 (IL18) modulates immune functions by inducing interferon–Î ³(IFN-ÃŽ ³) production and promoting Th1 immune responses. In the present study I amplified and cloned the sequence (582 bp) encoding full length bovine IL18 from peripheral blood mononuclear cells (PBMC) stimulated with Phytohaemoglutinin (PHA). Nucleotide and the deduced amino acid sequence of the cloned IL18 showed an identity of 86-98% with IL 18 sequences of the other ruminants compared. The insert was sub cloned in to eukaryotic expression vector (PcDNA) .The specificity of the expressed IL 18 was confirmed by western blotting. The insert was sub cloned in to pET 32a vector and expressed in E.Coli as fusion protein of 42kDa. The specificity of the expressed IL 18 was confirmed by western blotting. The biological activity of the purified protein was analysed for its ability to induce IFN-ÃŽ ³ production in PBMC as measured by Enzyme linked immunosorbent assay (ELISA) and quantitative polymerase cha in reaction (qPCR). IL18 anti FMD viral activity was conformed in vitro in BHK-21 cells by using plaque assay; viral replication was quantified by Real time PCR, ELISA and titration assays. 3. Study of the expression of the constructs in vitro in BHK-21 Cells Expression of cloned P12A3C and IL18 genes were studied in mammalian expression system for confirming the frame and intactness. The P12A3C, IL18 genes cloned under Eukaryotic promoter was transfected in BHK 21 cells with lipid based lipofectamine. Subsequently, the proteins were confirmed by Western blotting by using using anti FMDV serotype ‘Asia’, serum from experimentally infected cattle. IL18 transfected cell lysate showed 18 KDa by using human IL18 Mab. 4. PLG microparticles preparation and characterization The PLG/CTAB micro particles were prepared using a solvent evaporation technique essentially as described previously and briefly, the micro particles were prepared by emulsifying 10ml of a 6% (w/v) polymer solution in methylene chloride with 1ml of TE buffer at high speed using an soniprep. The primary emulsion was then added to 50ml of distilled water containing CTAB (0.5%, w/v). This resulted in the formation of a water/oil/water emulsion which was stirred at 6000rpm for 12h at room temperature, allowing the methylene chloride to evaporate. The resultingmicro particles were washed in distilled water by centrifugation at 10,000 Ãâ€" g and freeze dried. The plasmid construct was adsorbed onto the microparticles by incubating 100 mg of cationic microparticles with 100 mgs (1 mg/ml solution) of plasmid DNA at 40C for 6 h. The coated microparticles were then separated washed with TE and freeze-dried. Amount of plasmid adsorbed on PLG particles was quantified by eluting the DNA by 0.2 N NaOH (incubation for 10 h at 4 0C and measuring the Optical Density (OD) at 260 nm. Blank PLG micro particles controls were run simultaneously to deduct background value. The size distribution of the micro particles was determined using a particle size analyzer and electron microscopy. 5.A. Evaluation of the Immunological response of various DNA vaccine constructs in guinea pigs. Foot and Mouth Disease (FMD) can be controlled by regular vaccination and restricting the movement of animals infected in the endemic countries.. DNA vaccine construct was made with P1-2A3C coding sequences of serotype Asia1 in p C DNA. To evaluate the optimal dose of the construct in guinea pigs, the plasmid was coated on cationic Poly Lacto-co-Glycolide (PLG) micro particles was injected in to guinea pigs at 2,5,10,15,20,30 ug doses intramuscularly. Sera samples collected from the vaccinated animals at 21st dpv were evaluated for immune response by Enzyme linked immunosorbent assay (ELISA), Serum neutralization test (SNT) and MTT assay. Maximum ELISA / SNT titers and MTT stimulation indices were observed at 10  µg dose which also gave 83% protection when the guinea pigs were challenged with homologues virus. 10ug was found to be the optimal dose to guinea pigs. P12A3CpCDNA and bovine IL-18 pcDNA plasmids were constructed under CMV promoter and the coated with Cationic PLG microparticle, immune response of the co administered constructs was evaluated in guinea pigs. Both the plasmids constructed under CMV promoter and 10 µgs each of the plasmids were inoculated intra muscularly in guinea pigs with a booster dose at 21st day post vaccination (dpv). Both humoral and cellular immune response were analysed by IgG1, IgG2 enzyme linked immunosorbent assay (ELISA), Serum neutralization test (SNT) and MTT assay. Th1, Th2 cytokine profile was analysed by real time PCR and the phenotyping of T cell sub population in the peripheral blood was performed by flowcytometry. The results have sown significantly higher humoral and cell mediated immune responses in P12A3CIL18+PLG group than P12A3C IL18, and inactivated virus vaccine inoculated groups. Similarly, higher CD4, CD8 population and Th1, Th2 cytokine levels were seen in former group. P12A3CIL18+PLG vaccine protected all the six animals when challenged with homologous virus compared to five in inactivated virus vaccine group respectively. These results have shown that the plasmid encoding for P12A3C pcDNA when co inoculated with IL18 and PLG induce higher and protective immune responses, suggesting rBoIL-18 and Micro particles has a potential to enhance the efficacy of vaccine against FMD. 5. B Evaluation of the Immunological response of various DNA vaccine constructs in Cattle. Healthy male cattle calves of local breed ( Hallikar Breed) of 6 months to one year age group were purchased from local village shandy( cattle market). These animals were housed in healthy animal shed facilities available at IVRI Animal experimental station at Yelahanka , Bangalore. After initial quarantine the animals were bled and the sera were screened for FMDV antibodies for serotype Asia 1 by SNT. The FMD antibody free animals were divided in to 6 groups of six animals each namely Group I to Group IV. All the group were vaccinated with each construct with 200 ug injected by intramuscularly except conventional vaccine group injected with 2 ml of FMDV Inactivated vaccine. One group kepted for control group (vaccinated with PBS) .After 21 st days of first vaccination with same amount booster dose was injected P12A3CpCDNA and bovine IL-18 pcDNA plasmids were constructed under CMV promoter and the coated with Cationic PLG microparticle, immune response of the co administered constructs was evaluated in guinea pigs. Both the plasmids constructed under CMV promoter and 200 µgs each of the plasmids were inoculated intra muscularly in calves with a booster dose at 21st day post vaccination (dpv). Both humoral and cellular immune response were analysed by IgG1, IgG2 enzyme linked immunosorbent assay (ELISA), Serum neutralization test (SNT) and MTT assay. Th1, Th2 cytokine profile was analysed by real time PCR (ÃŽ ³IFN, IL4, IL2, ÃŽ ±IFN, IL12, IL25,TLR-4,TLR3,TLR-2,IL8,IL10) and the phenotyping of T cell sub population (CD4 and CD8) and intracellular cytokine molecules (ÃŽ ³IFN, IL4, IL2) in the peripheral blood was performed by flowcytometry. The results have sown significantly higher humoral and cell mediated immune responses in P12A3CIL18+PLG group than P12A3C IL18, and inactivated virus va ccine inoculated groups. Similarly, higher CD4, CD8 population and Th1, Th2 cytokine levels were seen in former group. P12A3CIL18+PLG vaccine protected four out of six animals when challenged with homologous virus compared to 3 in inactivated virus vaccine group respectively. Non structural proteins,ELISA conformed in challenged animals.These results have shown that the plasmid encoding for P12A3C pcDNA when co inoculated with IL18 and PLG induce higher and protective immune responses, suggesting rBoIL-18 and Micro particles has a potential to enhance the efficacy of vaccine against FMD Journal papers and conference/seminar papers from Doctoral research work 1. Expression of Bovine (Bos indicus) interleukin-18 inEscherichia coli and its biological activity.Kotla Siva Reddy, Dowlathabad. Muralidhar Rao, Hosur Joyappa Dechamma,Veluvarthy V.S. Suryanarayana and Golla Ramalinga Reddy.Published in Microbiology and Immunology 2010; 54: 564–567. 2. Enhancement of DNA vaccine (P12A3C-pcDNA) efficacy against Foot- andMouth Disease by co-administration of Interleukin-18 expressing (IL18pcDNA) plasmid in Guinea Pigs. Siva Reddy .K. Muralidhar Rao.D., Badrinaryana.M. Suryanaryana.VVS. and Reddy G.R. Accepted in FEMS Immunology and Medical Microbiology. Dec -2010 1–9. 3. Dose optimization of Cationic PLG micro particle coated DNA vaccine against Foot and Mouth Disease in Guinea pigs. Siva Reddy, K.,Rashmi., B.R., Muralidhar Rao, D., Dechamma H.J., Banumathi .N., Suryanarayana V.V.S and Reddy .G.R. accepted in J.of Life science.(Article in press) 4. Cytokine profile studied by Real time PCR in FMDV antigen stimulated Bovine PBMC cells. Siva Reddy .K., Muralidhar Rao,D.,PrabhuDas,K., Suryanaryana.VVS., Reddy ,G.R. Accepted in Journal of Biotechnology , Bio engineering and Bio –Informatics. (Article in press). 5. Bos indicus Interleukin 18 complete coding sequence published in NCBI Gen bank .SivaReddy,K., Muralidhar Rao,D., Dechamma,H., Banumathi,N.,Suryanaryana,V. and Reddy,G. Acc.No. FJ985771 6. Enhancement of DNA vaccine (P12A3C-pcDNA) efficacy against Foot- andMouth Disease by co-administration of Interleukin-18 expressing (IL18pcDNA) plasmid in Guinea Pigs. Siva Reddy .K. Muralidhar Rao.D., Badrinaryana.M. Suryanaryana.VVS. and Reddy G.R. Presented in Society for applied biotech biotechnology (SAB) annual conference at Dharmapuri Dec 17,18 7.Bovine Interleukin -18 inhibits Foot-and-Mouth Disease virus Replication in BHK- 21 cells. K. Siva Reddy, D.Murali Dhar Rao, Kakoli Ahmed, H.J Dechamma N.Bhanumathi ,VVS Suryanarayana ,G.R Reddy presented at VIROCON 2010 XIX National Conference RECENT TRENDS IN VIRAL DISEASE PROBLEMS AND MANAGEMENT SVU Tirupathi, Mar 18-20 ,2010 . 8. Cationic Micro Particle (PLG) coated DNA vaccination Induces a long term immune response and Protective Immunity against Foot –and-Mouth disease virus. K. Siva Reddy, Rashmi Dechamma N.Bhanumathi ,VVS Suryanarayana ,G.R Reddy Presented at VIROCON 2010 XIX National Conference RECENT TRENDS IN VIRAL DISEASE PROBLEMS AND MANAGEMENT SVU Tirupathi, Mar 18-20 ,2010. 9. Dose response studies of ID- p VAC (SECRETORY VECTOR CONSTRUCT) coated on cationic PLG micro particles against FMDV in guinea pigs. Siva Reddy K., Reddy G.R. Presented at SBC Annual conference Impact of Basic and Translational Research on Medicine, Agriculture and Industry, IIT Madras 18-20 DEC -2008. Communicated Articles 1. Cationic Micro Particle (PLG) coated DNA vaccination induces a long term immune response and Protective Immunity against FMD in GuineaPigs. Siva Reddy ,K ., MuraliDhar Rao,D.,Rashmi, B.R., Dechamma H,J., Banumathi.,N., Suryanarayana V.V.S and Reddy G.R Communicated in to Vet Immunology and Immunopathology(Under review). 2. Bovine Interleukin 18 inhibits Foot and mouth disease virus replication in BHK-21 Cells. K. Siva Reddy, D.MuraliDhar Rao, K.PrabhuDas, VVS Suryanarayana,G.R Reddy communicated in to Biotechnology and Applied Biochemistry. 3. Enhanced efficacy of a Foot and mouth disease DNA vaccine (P12A3CpcDNA) by adsorption onto cationic PLG microparticle in guinea pigs .K. Siva Reddy, D.MuraliDhar Rao, K.PrabhuDas, VVS Suryanarayana ,G.R Reddy communicated into International journal of Immunopharmacology.

Project Management – the Importance of the Planning Process

An individual assignment under the topic: „Project Management – the Importance of the Planning Processâ€Å" Teacher: Maria do Sacramento Basilio Student: 2012 Beja Content Content2 Introduction4 Project Management5 The Life Cycles of Projects7 PLANNING PROCESS8 Project Plan Elements11 Project Control13 Project Termination14 Conclusion16 Literature17 Introduction Each organization‘s activity in its own way contributes to organization‘s goals. It is not always easy to assess the impact of the work or the decision to achieve these objectives in the context of a complex organization of activities.Common to assume that what unites all the organizations processes to the general population and focus their efforts on a defined term direction, is a strategy. Project work is one of the extraordinary management forms. Each project is progressing to a certain stage, which is called the project life cycle. Despite the widely different names can be said that all projects are characterized by four main phases: initiation, planning, realization, finishing. An object of the Project: the planning process. The aim: to analize the essential part of Project life cycle – planning process.My essay will consists of two parts. In the first part I am going to introduce the Project Management and Project Life Cycle, in the second part of the project I will analyse the planning process and will make a conclussion about it‘s importance. Project Management In order to understand project management, one must begin with definition of a project. A project can be considered to be any series of activities and tasks that: * Have a specific objective to be completed within certain specifications * Have defined start and end dates * Have funding limits Consume resources (money, people, equipment) Project management, on the other hand, involves project planning ant project monitoring and includes such items as: Project planning: * Definition of work requiremen ts * Definition of quantity and quality work * Definition of resources needed Project monitoring: * Tracking progress * Comparing actual outcome to predicted outcome * Analyzing impact * Making adjustments Successful project management can then be defined as having achieved the project objectives: * Within time * Within cost * At the desired performance/technology level While utilizing the assigned resources effectively and efficiently * Accepted by the customer The potential benefits from project management are: * Identification of functional responsibilities to ensure that all activities are accounted for, regardless of personal turnover * Minimizing the need for continuous reporting * Identification of time limits for scheduling * Identification of methodology for trade-off analysis * Measurement of accomplishment against plans * Early identification of problems so that corrective action may follow * Improved estimating capability for future planning Knowing when objectives canno t be met or will be exceeded Unfortunately, the benefits cannot be achieved without overcoming obstacles such as: * Project complexity * Customer’s special requirements * Organizational restructuring * Project risks * Changes in technology * Forward planning and picking Project management can mean different things to different people. Quite often, misunderstand the concept because they have ongoing projects within their company and feel that they are using project management to control these activities.In such a case, the following might be considered an appropriate definition: Project management is the art of creating the illusion that any outcome is the result of a series of predetermined, deliberate acts when, in fact, it was dumb luck. Although this might be the way that some companies are running their projects, this is not project management. Project management is designed to make better use of existing resources by getting work to flow horizontally as well as verticall y within the company.This approach does not really destroy the vertical, bureaucratic flow of work but simply requires that line organizations talk to another horizontally so work will be accomplished more smoothly throughout the organization. The vertical flow of work is still the responsibility of the line managers. The horizontal flow of work is the responsibility of the project managers, and their primary effort is to communicate and coordinate activities horizontally between the line organizations.The following would be an overview definition of project management: Project management is the planning, organizing, directing, and controlling of company resources for a relatively short-term objective that has been established to complete specific goals and objectives. Furthermore, project management utilizes the systems approach to management by having functional personnel (the vertical hierarchy) assigned to a specific project (the horizontal hierarchy). The above definition requi res further comment. Classical management is usually considered to have five functions or principles: * Planning Organizing * Staffing * Controlling * Directing In the above definition, the staffing function has been omitted. This was intentional because the project manager does not staff the project. Staffing is line responsibility. The project manager has the rigt to request specific resources, but the final decision of what resources will be committed rests with the line managers. Talking about the meaning by a â€Å"relatively† short-term project, not all industries have the same definition for a short-term project.In engineering, the project might be for six months or two years; in construction, three to five years; in nuclear components, ten years; and in insurance, two weeks. Long-term projects, which consume resources full-time, are usually set up as a separate division (if large enough) or simply as a line organization. The Life Cycles of Projects All organisms have a life cycle. They born, grow, wane, and die. This is true for all living things, for stars and planets, for the products we buy and sell, for our organizations, and for our projects as well.A project’s life cycle measures projects completion as a function of either time (schedule) or resources (budget). This is life cycle must be understood because the PM’s managerial focus subtly shifts at different stages of the cycle. During the early stages, the PM must make sure that the project plan really reflects the wishes of the client as well as the abilities of the project team and is designed to be consistent with the goals and objectives of the parent firm. As the project goes into the implementation stage of its life cycle, the PM’s attention turns to the job of keeping the project on budget nd schedule – or, when chance interferes with progress, to negotiating the appropriate trade-offs to correct or minimize the damage. At the end of the project, the PM turns into a â€Å"fuss-budget† to assure that the specifications of the project are truly met, handling all the details of closing out the books on the project, making sure there are no loose ends, and that every â€Å"i† is dotted and â€Å"t† crossed. Many projects are like building a house. A house-building project starts slowly with a lot of discussion and planning. Then construction begins and progress rapid.When the house is built, but not finished inside, progress appears to slow down and it seemingly takes forever to paint everything, to finish all the trim, and to assemble and install the built-in appliances. Progress is slow-fast-slow, as shown in figure: It used to be thought that S-shaped curve of figure represented the life cycle for all projects. While this is true of many projects, there are important exceptions. This is about the project that approaches completion by a very different route that the tradidtional S-curve, as shown in this figure:T o make a conclusion of the topic about product life cycle, there are two different paths (life cycles) along which projects progress from start to completion. One is S-shaped and the other is J-shaped. It is an important distinction because identifying the different life cycles helps the PM to focus attention on appropriate matters to ensure successful project completion. PLANNING PROCESS The most important responsibilities of a project manager are planning, integrating and executing plans. Almost all projects, because of their relatively short duration and often prioritized control of resources, require formal, detailed planning.The integration of a planning activities is necessary because each functional unit may develop its own planning documentation with little regard for other functional units. Planning, in general, can be best described as the function of selecting the enterprise objectives and establishing the policies, procedures, and programs necessary for achieving them. P lanning in a project environment may be described as establishing a predetermined course of action within forecasted environment. The project’s requirements set the major milestones, and the line managers hope that they can meet them.If the line manager connot commint because the milestones are perceived as unrealistic, the project manager may have to develop alternatives, one of which may be to move the milestones. Upper-level management must become involved in the selection of alternatives during the planning stage. Planning is, of course, decision making, since it involves choosing among alternatives. Planning is a required management function to facilitate the comprehension of complex problems involving interacting factors. The project manager is the key to successful project planning.It is desirable that the project manager be involved from project conception through execution. Project planning must be systematic, flexible enough to handle unique activities, disciplined through reviews and controls, and capable of accepting multifunctional inputs. Successful project managers realize that project planning is an iterative process and must be performed throughout the life of the project. One of the objectives of project planning is to completely define all work required (possibly through the development of a documented project plan) so that will be readily identifiable to each project participant.This is necessity in a project environment because: * If the task is well understood prior to being performed, much of the work can be preplanned. * If the tas is not understood, the during the actual task execution more knowledge is gained that, in turn, leads to changes in resource allocations, schedules, and priorities. * The more uncertain the task, the greater the amount of information that must be processed in order to ensure effective performance. Without proper planning, programs and projects can start off â€Å"behind the eight ball† because o f poorly defined requirements during the initial planning phase.Below is a list of the typical consequences of poor planning: * Project initiation * Wild enthusiasm * Disillusionment * Chaos * Search for the guilty * Punishment of the innocent * Promotion of the nonparticipants * Definition of the requirements Obviously, the definition of the requirements should have been the first step. There are four basic reasons for project planning: * To eliminate or reduce uncertainty * To improve efficiency of the operation * To obtain a better understanding of the objectives * To provide a basis for monitoring and controlling work Planning is decision making based upon futurity.It is a continuous process of making entrepreneurial decisions with an eye to the future, and methodically organizing the effort needed to carry out these decisions. Furthermore, systematic planning allows an organization of set goals. The alternative to systematic planning is decision making based on history. This ge nerally results in reactive management leading to crisis management, conflics management, and fire fighting. Effective total program planning cannot be accomplished unless all of the necessary information becomes available ant project initation. These information requirements are: * The statement of work (SOW) The project specifications * The milestone schedule * The work breakdown structure (WBS) The statement of work (SOW) is a narrative description of the work to be accomplished. It includes the objectives of the project, a brief description of the work, the funding constraint if one exists, and the specifications and schedule. The schedule is a â€Å"gross† schedule and includes such things as the: * Start date * End date * Major milestones * Written reports The last major item is the work breakdown structure. The WBS is the breaking down of the statement of work into smaller elements so that better isibility and control will be obtained. Project Plan Elements The project master plan should contain nine elements: a project overview, a statement of objectives, a description of the technical and managerial approaches to the work, all contractual agreements, schedules of activities, a list of resource requirements or a project budget, personnel requirements, project evaluation methods, and preparations to meet potential problems. These are the elements that constitute the project plan and the basis for a more detailed planning of the budgets, schedules, work plan and general management of the project.Once this basic plan is fully developed and approved, it is disseminated to all interested parties. I would like to describe each element. * Overview. This is a short summary of the objectives and scope of the project. It is directed to top management and contains a statement of the goals of the project, a brief explanation of their relationship to the firm’s objectives, a description of the managerial structure that will be used for the project, an d a list of the major milestones in the project schedule. * Objectives. This contains a more detailed statement of the general goals noted in the overview section.The statement should include profit and competitive aims as well as technical goals. * General approach. This section describes both the managerial and the technical approaches to the work. The technical discussion describes the relationship of the project to available technologies. For example, it might note that this project is an extension of work done by the company for an earlier project. The subsection on the managerial approach takes note of any deviation from routine procedure – for instance, the use of subcontractors for some parts of the work. * Contractual aspects.This critical section of the plan includes a complete list and description of all reporting requirements, customer-supplied resources, liaison arrangements, advisory committees, project review and cancellation procedures, proprierty requirements , any specific management agreements, as well as the technical deliverables and their specifications, delivery schedules, and a specific procedure for changing any of the above. Completeness is a necessity in this section. If in doubt about whether an item should be included or not, the wise planner will include it. * Schedules.This section outlines the various schedules and lists all milestone events. The estimated time for each task should be obtained from those who will do the work. The project master schedule is constructed from these inputs. The responsible person or department head should sign off on the final, agreed-on schedule. * Resources. There are two primary aspects to this section. The first is the budget. Both capital and expense requirements are detailed by task, which makes that a project budget. One-time costs are separated from recurring project costs. Second, cost monitoring and control procedures should be described.In addition to the usual routine elements, the monitoring and control procedures must be designed to cover special resource requirements for the project, such as special machines, test equipment, laboratory usage or construction, logistics, field facilities, and special materials. * Personnel. This section list the expected personnel requirements of the project. Special skills, types of training needed, possible recruiting problems, legal or policy restrictions on work force composition, and any other special requirements, such as security clearances, should be noted here.It is helpful to time-phase personnel needs to the project schedule. This makes clear when the various types of contributors are needed and in what numbers. These projections are an important element of the budget, so the personnel, schedule, and resources sections can be cross-checked with one another to ensure consistency. * Evaluation Methods. Every project should be evaluated against standards and by methods established at the project’s inception. T his section contains a brief description of the procedure to be followed in monitoring, collecting, storing, and evaluating the history of the project. Potential Problems. Sometimes it is difficult to convince planners to make a serious attempt to anticipate potential difficulties. One or more such possible disasters such as subcontractor default, technical failure, strikes, bad weather, sudden required breakthroughs, critical sequences of tasks, tight deadlines, resource limitations, complex coordination requirements, insufficient authority in some areas, and new, complex, or unfamiliar tasks are certain to occur. The only uncertainties are which ones will occur and when.In fact, the timing of these disasters is not random. There are times, conditions, and events in the life of every project when progress depeneds on subcontractors, or the weather, or coordination, or resource availability, and plans to deal with unfavorable contingencies should be developed early in the projectâ⠂¬â„¢s life cycle. Some PMs disdain this section of the plan on the grounds that crises cannot be predicted. Further, they claim to be very effective firefighters. It is quite possible that when one finds such a PM, one has discovered an arsonist.No amount of current planning can solve the current crisis, but preplanning may avert some. Project Control The two fundamental objectives of control are: 1. The regulation of results through the alteration of activities. 2. The stewardship of organizational assets. Most discussions of the control function are focused on regulation. Physical Asset Control Physical asset control requires control of the use of physical assets. It is concerned with assets maintenance, whether preventive or corrective.At issue also is the timing of maintenance or replacement as well as the quality of maintenance. Physical inventory, whether equipment or material, must also be controlled. It must be received, inspected, and possibly stored prior to use. Records of all incoming shipments must be carefully validated so that payment to suppliers must also be applied to suppliers from inside the organization. Even such details as the project library, project coffee maker, project room furniture, and all the other minor bits and pieces must be counted, maintained, and conserved.Human Resource Control Stewardship of human resources requires controlling and maintaining the growth and development of people. Projects provide particulary fertile ground for cultivating people. Because projects are unique, differing one from another in many ways, it is possible for people working on projects to gain a wide range of experience in a reasonably short time. Measurement of physical resource conservation is accomplished through the familiar audit procedures. The measurement of human resource conservation is familiar audit procedures.The measurement of human resource conservation is far more difficult. Such devices as employee appraisals, personnel performan ce indices, and screening methods for appointment, promotion, and retention are not particularly satisfactory devices for ensuring that the conservation function is being properly handled. The accounting profession has worked for some years on the development of human resource accounting, and while the effort has produces some interesting ideas, human resource accounting is not well accepted by the accounting profession.Financial Resource Control Though accountants have not succeeded in developing acceptable methods for human resource accounting, their work on techniques for the conservation of financial resources has most certainly resulted in excellent tools for financial control. This is the best developed for the basic areas needing control. It is difficult to separate the control mechanisms aimed at conservation of financial resources from those resources from those focused on regulating resource use. Most financial controls do both.Capital investment controls work to conserve the organization’s assets by insisting that certain conditions be met before capital can be expended, and those same conditions usually regulate the use of capital to achieve the organization goal of a high return on investments. The techniques of financial control, both conservation and regulation, are well known. They include current assets controls, and project budgets as well as capital investment controls. These controls are exercised through a series of analyses and audits conducted by the accounting function for the most part.Project Termination As it must to all things, project termination comes to every project. At times, project death is quick and clean, but more often it is a long process. The process of termination is never easy, always complicated, and, as much as we might wish to avoid it, almost always inevitable. The problem is how to accomplish one of the several levels of what is meant by project termination with a minimum of trouble and administrative dislo cation. A project can be terminated in one of four ways: * by extinction * addition * integration * starvation y Extinction: * The project has successfully completed scope and the client has accepted it. * It has been superseded by the external developments like technological advancement, market crisis etc * It has failed to achieve it’s goal. * It has no longer support from the Senior Management. It is also sometimes referred to as â€Å"termination by murderâ€Å". The important point to notice is that all project activity ceases in this kind of termination. by Addition: * The project is a major success. It becomes the formal part of the parent organization. The transition or   transfer of the resources such as the project personnel, materials and equipment to the newly created unit within the parent   organization. by Integration: * The project is successfully completed. The   project product is integrated to the operations of the client. This is the most common mo de and most complex operation. The resources are   released   and   disturbuted in the parent organization. by Starvation: * The project is terminated by budget decrement. * It is also known as   withdrawal of â€Å"life support†. The reason of this termination is generally to shadow the failure of non-accomplishment of the goals.This can save face of the senior management and avoid embarrassment. The Project Final Report incorporates the process knowledge gained from the Project. In addition to preservation of Project records, the Final Report embodies the experience from which we learn. It should include: Project performance comments, administrative performance comments, organizational structure comments, personnel suggestions, possibly a confidential section. Conclusion Project planning is probably the most time-consuming project management activity.Project planning continues from the initial idea through to the system delivery. Plans must be regulary updated using the new available information. There are many techniques for developing a project plan. They are fundamentally similar. All of them use a systematic analysis to identify and list the things that must be undertaken in order to achieve the project‘s abjectives, to test and validate the plan and to deliver it to user. Planning is an essential function in the success of any project. Planning does not refer simply to pulling out a calendar and recording things to do on random dates.Planning is all about actively plotting a course to meeting goals. Goals are really the start of any discussion about planning in a successful project. Literature 1. Harold Kerzner „Project Management. A Systems Approach to Planning, Scheduling, and Controllingâ€Å" sixth edition 2. Samuel J. Mantel Jr. Jack R. Meredith, Scott M. Shafer, Margaret M. Sutton „Project Management in Practiseâ€Å" second edition 3. Jack R. Meredith, Samuel J. Mantel „Project Management. A Managerial app roachâ€Å" third edition 4. V. Buda, A. Chmieliauskas „Projektu valdymasâ€Å" 2006

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This age is called the age of science, yet' we can’t deny the importance of literature in our lives. Literature is called the mirror of life; it is also called a mode of the expression of feelings and emotions. As long as human beings do have emotions and feelings, literature will be created and literature will be read. It gives a way to one's thought, modifies and brings new dimensions. It relaxes you when you are tense' enlightens your dark mind and lightens your heavy heart. This age is called the age of science, yet' we can’t deny the importance of literature in our lives.Literature is called the mirror of life; it is also called a mode of the expression of feelings and emotions. As long as human beings do have emotions and feelings, literature will be created and literature will be read. It gives a way to one's thought, modifies and brings new dimensions. It relaxes you when you are tense' enlightens your dark mind and lightens your heavy heart. â€Å"Roselilyâ₠¬  by Alice Walker, is one of the literatures that I really liked. In the short story Roselily, Alice Walker tells two stories in one.The most obvious story is the one about the Black American woman Roselily, who stands before the alter, just about to marry a muslim, while she thinks about her past, wonders about the future and is questioning whether she is making the right choice. The other, hidden story is the story about Black American women in general, their history and their ongoing search for something better. The way I understand the short story, Roselily`s story is, as it is presented to the reader through Roselily’s thoughts as she is in the middle of her wedding, a reflection of Black Americans` and women in particular situation around the 1960s.At this time, Blacks are free Americans with the equal rights as other Americans, in theory. Roselily is an independent woman of her time, but being a single mother of four children, working long hours for most likely lousy wages in a sewing plant, she is far from free. The Blacks are no longer slaves in the cotton fields, they are now paid slaves in the refinement industry. Roselily is most aware of her situation, and she is willing to leave her past and start a new life with a new man. She has probably been searching for a better life for quite some time, by being with different men, who all could give her a child, but not a new life.I am sensing an urge in Roselily, to move on, symbolized by all the cars described in the short story: They are constantly moving from one place to another, they give you mobility, prevent you from being stuck somewhere you do not want to be. Roselily knows that she does not want to stay in the sewing plant, she knows that she wants to move on to something better, but she does not know what better is, and she certainly doubts if what she has chosen will be better than what she had. Her divided personality is like the different groups of Blacks in the civil rights moveme nts.Some Blacks wanted segregation, some wanted their own nation, some wanted to be more African, some wanted to live like the White Americans, some were Muslims and some were Christians. They all agreed that their current situation was not acceptable, but they did not know how to improve it. At the same time as Roselily wants to start a new life, she is afraid of losing her roots. She pictures her children  «exalted on a pedestal, a stalk that has no roots », and  «She wonders how to make new roots ».Alice Walker here describes Black Americans and probably also her own search for identity. Which heritage is the right one; the one from their Black American ancestors, or the one from the African ancestors? As Roselily is about to marry, about to go to Chicago and start a new life with her husband, she is starting to regret her decision and faces the truth: Just like Black Americans went from the slavery in the cotton fields to paid slavery in the sewing plants, she is now mov ing to the slavery of Islam. She thinks of his religion and sees ropes, chains, handcuffs.She thinks of Chicago, her new home, and realizes that all she knows about the place is Lincoln, the president. President Lincoln abolished the slavery, but that did not free the Black Americans. Alice Walker shows feminism in this literature. Most of her works depicts racism, sexism, feminism, troubled relationships, and isolation. Alice Walker was the eighth child of sharecroppers. Despite the economic hardships of her family, she was remarkably dedicated to her education and graduated with degrees from both Sarah Lawrence and Spelman College.While attending school, Walker became frustrated with the lack of literature on the culture and history of the black experience, so she challenged educational institutions to create a representative curriculum. In the 1960s, she became involved in the civil rights movement. Her experiences became the basis for her excellent novel Meridian. Her best-known work, however, is The Color Purple. Critics and audiences alike have praised its richly drawn female characters and seemingly effortless use of black vernacular. Although she has written six novels, Walker remains very active politically, championing women’s issues and women’s work.

The Epic of Gilgamesh Ancient Mesopotamian - Free Essay Example

Sample details Pages: 2 Words: 557 Downloads: 10 Date added: 2019/07/29 Category History Essay Level High school Tags: Gilgamesh Essay Did you like this example? In terms of religion, the epic tells us that the Mesopotamian people are polytheistic; they believed each of the gods to be the supreme authorities of their respective domains. They also saw them as their protectors which can be seen at the very beginning of the tale when they cry to the gods about their problem with Gilgamesh as a ruler effectively asking for help. Because they looked to the gods for protection one can infer they were deeply religious; much like the religions of today, people will be loyal to whichever god they feel will protect them evil during their lives.. Don’t waste time! Our writers will create an original "The Epic of Gilgamesh Ancient Mesopotamian" essay for you Create order Although there is not much said in the way of gender roles in ancient Mesopotamia, they seemed to be believers in autonomy, as evidenced by one of their main grievances of Gilgamesh; he was indiscriminately raping of Uruks women without regard to their virginity or marital status basically, the women had no choice. The people of Mesopotamia ultimately wanted to live in peace and harmony under a king who genuinely cared about their wellbeing instead of his own. When complaining about Gilgamesh they remark that a king should be somewhat of a shepherd to his people which indicates they would like their king to be guiding and nurturing to them, as a shepherd is someone who raises and cares for sheep. Discussion Response This is an articulate and well-thought response. I would agree that the people of Uruk favored civilization over wilderness as evidenced by the Mesopotamian people and their desires for the type of king theyd like, wise, comely, and resolute (Sandars) all traits of a civilized demeanor. Their favor of civilization over savagery is made interesting because, in the end, it was the savagery of Enkidu that played a significant role in changing the attitude of Gilgamesh and changing him into the type of ruler they wanted and needed. Discussion Prompt Two One choice that Odysseus made that demonstrates his strong leadership quality is his decision to try and deceive the Cyclops Polyphemus; this is an incredible decision not only because of Polyphemus physical nature but because Odysseus had just seen him brutally murder and consume two of his men. Instead, he kept his cool and demonstrated the mental tenacity of a leader which allowed for him to create his successful plan to get Polyphemus drunk, give him the wrong name of Nobody so he could not effectively ask the other Cyclopes for help, Nobody is killing me now by fraud and not by force! (Homer 9.450), and then escape under the animals. Another decision made by Odysseus that shows his leadership ability is his choice be dedicated to Athenas plan for him to stay disguised as a beggar when he returns to Ithaca. His leadership skills also help to keep up the rouse when a nurse recognizes him due to a scar he has as he is able to convince her to keep quiet. Even after seeing his wife Penelope and being tempted to reveal himself, he was able to put his personal feelings aside for the good of his family and the people of Ithaca. Effective leaders all have certain qualities in common including a commitment to their causes, the ability to persevere under adverse circumstances, and a willingness to take risks to achieve desired resultsall characteristics shown by Odysseus.