## Research Paper Writing

Chapter 13

11-  Implement the linear optimization model that you developed for Valencia Products in Problem 4 on a spreadsheet and use Solver to find an optimal solution. Interpret the Solver Answer Report, identify the binding constraints, and verify the values of the slack variables by substituting the optimal solution into the model constraints.

12.Implement the linear optimization model that you developed for ColPal Products in Problem 5 on a spreadsheet and use Solver to find an optimal solution. Interpret the Solver Answer Report, identify the binding constraints, and verify the values of the slack variables by substituting the optimal solution into the model constraints.

1. 13.Implement the linear optimization model that you developed for Burger Office Equipment in Problem 6 on a spreadsheet and use Solver to find an optimal solution. Interpret the Solver Answer Report, identify the binding constraints, and verify the values of the slack variables by substituting the optimal solution into the model constraints.
2. 14.Implement the linear optimization model that you developed for the investment scenario in Problem 7 on a spreadsheet and use Solver to find an optimal solution. Interpret the Solver Answer Report, identify the binding constraints, and verify the values of the slack variables by substituting the optimal solution into the model constraints.
3. *15.Implement the linear optimization model that you developed for Bangs Leisure Chairs in Problem 8 on a spreadsheet and use Solver to find an optimal solution.
1. Interpret the Solver Answer Report, identify the binding constraints, and verify the values of the slack variables by substituting the optimal solution into the model constraints.
2. Suppose that Mr. Bangs wants to limit the number of Adirondack chairs to at most 20. Modify and re-solve your model to determine the new solution.
3. Suppose that Mr. Bangs does not want to spend more than 40 hours each month on any one activity. Modify and re-solve your original model to determine the new solution.
4. *16.Implement the linear optimization model that you developed for the Morton Supply Company in Problem 9 on a spreadsheet and use Solver to find an optimal solution. Interpret the Solver Answer Report and identify the binding constraints.

17- Implement the linear optimization model that you developed for Malloy Milling in Problem 10 on a spreadsheet and use Solver to find an optimal solution. Interpret the Solver Answer Report and identify the binding constraints.

How Solver Works

1. 18.For the Valencia Products model in Problem 4, graph the constraints and identify the feasible region. Then identify each of the corner points and show how increasing the objective function value identifies the optimal solution.
2. 19.For the ColPal model in Problem 5, graph the constraints and identify the feasible region. Then identify each of the corner points and show how increasing the objective function value identifies the optimal solution.

Chapter 14

Solve Problem 11 in Chapter 13 (Valencia Products) to ensure that the number of units produced is integer-valued. How much difference is there between the optimal integer solution objective function and the linear optimization solution objective function? Would rounding the continuous solution have provided the optimal integer solution?

2.Solve Problem 12 in Chapter 13 (ColPal Products) to ensure that the number of minutes of radio and TV ads is integer-valued. How much difference is there between the optimal integer solution objective function and the linear optimization solution objective function? Would rounding the continuous solution have provided the optimal integer solution?

*3.Solve Problem 15 in Chapter 13 (Bangs Leisure Chairs) to ensure that the number of units produced is integer-valued. How much difference is there between the optimal integer solution objective function and the linear optimization solution objective function? Would rounding the continuous solution have provided the optimal integer solution?

4.For the Brewer Services scenario described in this chapter, suppose that 11 permanent employees are hired. Find an optimal solution to minimize the number of part-time employees needed.

*5.The Gardner Theater, a community playhouse, needs to determine the lowest-cost production budget for an upcoming show. Specifically, they have to determine which set pieces to construct and which, if any, set pieces to rent from another local theater at a predetermined fee. However, the organization has only two weeks to fully construct the set before the play goes into technical rehearsals. The theater has two part-time carpenters who work up to 12 hours a week, each at \$10 an hour. Additionally, the theater has a part-time scenic artist who can work 15 hours per week to paint the set and props as needed at a rate of \$15 per hour. The set design requires 20 flats (walls), two hanging drops with painted scenery, and three large wooden tables (props). The number of hours required for each piece for carpentry and painting is shown below:

Flats, hanging drops, and props can also be rented at a cost of \$75, \$500, and \$350 each, respectively. How many of each unit should be built by the theater and how many should be rented to minimize total costs?

7.Joe is an active 26-year-old male who lifts weights six days a week. His rigorous training program requires a diet that will help his body recover efficiently. He is also a graduate student who is looking to minimize the cost of consuming his favorite foods. Joe is trying to gain weight, or at least maintain his current body weight, so he is not concerned about calories. His personal trainer suggests at least 300 grams of protein, 95 grams of fat, 225 grams of carbohydrates, and no more than 110 grams of sodium per day. His favorite foods are all items that he is familiar with preparing, as shown in the table Data for Problem 7. He is willing to consume multiple servings of each food per day to meet his requirements, although he cannot eat more than one steak per day and does not want to eat more than three pulled pork sandwiches a day. He needs to consume at least two servings of broccoli and one serving of carrots per day but is willing to eat two servings of carrots if necessary. Joe likes a certain brand of nutrition bars, but he would not eat more than one. Unless previously noted, he does not want more than five servings of any one food. How many servings of each food should he have in an optimal daily diet?

8- Gales Products manufactures ribbon for thermal transfer printing, which transfers ink from a ribbon onto paper through a combination of heat and pressure. Different types of printers use different sizes of ribbons. The company has forecasted demand for seven different ribbon sizes, as shown below.

The rolls from which ribbons are cut are 900 mm in length. Scrap is valued at \$0.07 per millimeter. Generate ten different cutting patterns so that each size can be cut from at least one pattern. Use your data to construct and solve an optimization model for finding the number of patterns to cut to meet demand and minimize trim loss.

10- Fuller Legal Services wants to determine how much time to allocate to four different services: business consulting, criminal work, nonprofit consulting, and wills/trusts. Mr. Fuller has determined the average hourly fees and the minimum and maximum hours (for consulting and criminal work) and cases (for wills/trusts) that he would like to spend on each. He has no shortage of demand for his services. The relevant data are shown in the table Data for Problem 10. Develop and solve an integer optimization model to maximize monthly revenue.

*11.Riesemberg Medical Devices is allocating next year’s budget among its divisions. As a result, the R&D Division needs to determine which R&D projects to fund. Each project requires various software and hardware and consulting expenses, along with internal human resources. A budget allocation of \$1,300,000 has been approved, and 35 engineers are available to work on the projects. The R&D group has determined that at most one of projects 1 and 2 should be pursued, and that if project 4 is chosen, then project 2 must also be chosen. Develop a model to select the best projects within the budget.

Chapter 15:

1. For the Valencia Products scenario (Problems 4 and 11 in Chapter 13), use the spreadsheet model to answer the following questions by changing the parameters and re-solving the model. Answer each question independently relative to the original problem.
1. If the unit profit for SpeedBuster is decreased to \$130, how will the optimal solution and profit change?
1. If the unit profit for LaserStop is increased to \$210, how will the optimal solution and profit change?
1. If an additional 1,500 units of component A are available, can you predict how the optimal solution and profit will be affected?
1. If a supplier delay results in only 3,000 units of component B being available, can you predict how the optimal solution and profit will be affected? Can you explain the result?
2. For the ColPal Products scenario (Problems 5 and 12 in Chapter 13), use the spreadsheet model to answer the following questions by changing the parameters and re-solving the model. Answer each question independently relative to the original problem.
1. Suppose that the exposure for TV advertising was incorrectly estimated and should have been 875. How would the optimal solution have been affected?
1. Radio listening has gone down, and new marketing studies have found that the exposure has dropped to 150. How will this affect the optimal solution?
1. The marketing manager has increased the budget by \$2,000. How will this affect the solution and total exposure?
3. For the Burger Office Equipment scenario (Problems 6 and 13 in Chapter 13), use the spreadsheet model to answer the following questions by changing the parameters and re-solving the model. Answer each question independently relative to the original problem.
1. If 25% of the pine is deemed to be cosmetically defective, how will the optimal solution be affected?
1. The shop supervisor is suggesting that the workforce be allowed to work an additional 50 hours at an overtime premium of \$18/hour. Is this a good suggestion? Why or why not?
1. If the unit profit for standard desks is increased to \$280, how will the optimal solution and total profit be affected?
1. If the unit profit of standard desks is only \$190, how will the optimal solution and total profit be affected?
4. For the Markowitz model in Example 14.10, determine how the minimum variance and stock allocations change as the target return varies between 8% and 12% (in increments of 1%) by re-solving the model. Summarize your results in a table, and create a chart showing the relationship between the target return and the optimal portfolio variance. Explain what the results mean for an investor.
5. Figure 15.32 shows the Solver Sensitivity Report after solving the Crebo Manufacturing problem in Chapter 13 (Example 13.10). Using only the information in the Sensitivity Report, answer the following questions.
1. Explain the value of the reduced cost (−0.3)(−0.3) for the number of plugs to produce.
2. If the gross margin for rails is decreased to \$1.05, can you predict what the optimal solution and profit will be?
3. Suppose that the gross margin for rivets is increased to \$0.85. Can you predict what the optimal solution and profit will be?
4. If the gross margin for clips is reduced to \$1.10, can you predict what the optimal solution and profit will be? What if the gross margin is reduced to \$1.00?
5. Suppose that an additional 500 minutes of machine capacity is available. How will the optimal solution and profit change? What if planned maintenance reduces capacity by 300 minutes?

## Research Paper Writing Help

PROTEIN PURIFICATION – A COMPUTER SIMULATION

A week ago, you read a paper in which various separation techniques were used.  This week, you will experiment with a computer program that simulates additional methods of separation.

Protein Purification was written by Andrew Booth of the University of Leeds as part of the eLABorate  project.  The project was funded under the Teaching and Learning Technology Programme by the four higher education funding bodies, HEFCE, HEFCW, SHEFC, and DENI.  © The University of Leeds, 1997.

There are two parts to this exercise. The first (Part I) is to be done before you arrive in lab through the following link:

http://www.agbooth.com/pp_ajax/

The protein purification in Part II is to be completed in lab, working individually.

To carry out this exercise, you must familiarize yourself with the program and the possible purification steps. Before coming to laboratory, complete Part I and review the “Written Work for Protein Purification Lab” found at the beginning of Part II.

Protein Purification

Aims

The aims of this computer simulation are:

to familiarize you with a range of protein separation techniques

to allow you to explore how these techniques work, and to see their constraints

to allow you to devise schemes to purify proteins from a mixture, using combinations of      techniques

How to use the program

Protein Purification is a Windows based program. It uses the standard menu bars and drop-down menus; these can be operated using the mouse or keyboard commands. This document assumes that you are reasonably familiar with Windows. If you have not used Windows before, ask a demonstrator for some help before you start. Throughout this document you will be given the necessary information to use each part of the program. If you follow the exercises in the order in which they are set, you will learn how to use the program as you progress.

Protein Purification has an extensive Help system including: more information about the various protein separation techniques; a list of the time costs of each of the techniques; clues about strategies that you might use; and a progress report of your current separation scheme. The Help menu is available from all parts of the program and you will also find Info buttons; press these to get instant help related to the task in hand.

When you want to finish using Protein Purification, choose Go home from the Quit menu. When you are asked if you want to store your material, answer No. (Remember that you will lose your work on the protein if you answer No, so choose Cancel to return to the program if you change your mind).

Tasks to perform using the program

The exercises are divided into two parts. Part I familiarises you with the program and allows you to explore the various protein separation techniques using a simple mixture of proteins. Once you are familiar with all of these methods, in Part II you will be asked to use combinations of separation techniques to produce a pure sample of a protein from a much more complex mixture.

The tasks in Part I require short answers to be completed on this sheet, preferably before coming to lab. You are strongly advised to read all the questions in a particular section before you begin working on the computer; this should save you time, as many of the questions may be answered from a single, carefully planned experiment. When you have completed Part I there will be a group discussion when you may be asked to present your answers to the rest of the class.

The report on your work in Part II will be completed in lab and handed in. You are strongly advised to follow the guidelines on presentation.

For Part I, complete the sections on (1) SDS-PAGE, (2) Ion-exchange chromatography, and (3) Gel filtration. Sections 5 – 7 on heat treatment, ammonium sulfate fractionation, and hydrophobic interaction chromatography may also be useful for your protein purifications in Part II, but they do not work with the protein mixture Easy3_Mixture (they do work for the more complicated mixture that you’ll be using in Part II). Isoelectric focusing (4) and affinity chromatography (8) also work, but you are not allowed to use them in your purifications!

PART I                     Protein Purification Techniques

Getting started

1. On the title page, click on Start and select Choose a Mixture (or Start from beginning) from the drop-down menu.

2. For the exercises in Part I choose the Easy3_Mixture.

3. Another dialog box pops up; this allows you to select the protein you wish to purify from the mixture by its number. For the first task choose protein 1.

4. A box appears with information about protein 1; when you have read it and noted the relevant information click OK.

5. Throughout the exercises in Part 1 you will need to keep returning to this menu to select another of the proteins from this simple mixture. To do this, from the Quit menu choose Abandon scheme and start again; then start again from the beginning.

It is a good idea before you begin the exercises, to explore the menus and to familiarise yourself with the program in general.

1)  SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis)

Introduction:SDS-PAGE is a separation technique used largely for analysis. It can be utilised to discover properties of the proteins that we wish to purify using other, more preparative, techniques. It also allows us to follow the progress of our protein purification. In preparing the protein sample for SDS-PAGE the proteins are denatured and SDS (which is negatively charged) is bound to the proteins in a constant mass ratio. The result is that all the proteins have almost a constant charge to mass ratio, and during electrophoresis will be separated solely on the basis of their size. An electric field is created across the polyacrylamide gel, and the negatively charged proteins migrate towards the anode (positive electrode). As they move through the gel, the larger molecules are retarded whereas the smaller molecules can pass more easily through the pores in the gel. The result of this sieving is that the smallest molecules would reach the anode first, and the largest last. However, the electric field is turned off before the proteins reach the end of the gel, and the mass of the proteins can be estimated from the distance travelled through the gel. To estimate the relative molecular mass (Mr) of the sample proteins, standard proteins of known Mr are run on the same gel to provide a reference. To reveal the proteins on the gel a stain, such as Coomassie blue, can be used. If the protein we are interested in has been isolated previously, a specific antibody may be available for it. If this is the case then the protein in question can be identified from the array of proteins in the mixture, using an immunoblot. Note that as the proteins are denatured prior to separation by SDS-PAGE, the Mr that can be determined is that of the individual subunits, and not of the native protein. If the protein only has one subunit then of course its Mr can be estimated directly using this technique. If a protein is composed of more than one type of subunit, then more than one spot can be seen for this protein on the gel.

It is often useful to be able to perform a 2-dimensional separation of the protein mixture. The first separation involves isoelectric focusing (IEF). IEF separates proteins on the basis of their pI (isoionic point), each protein migrates in an electric field through a pH gradient until it reaches a position where the pH of the surrounding buffer is equal to the pI of the protein (see section 4, p63 for more details). The IEF is performed in a rod gel (with no SDS present). Once the separation has been completed the rod gel is moulded to the top of a polyacrylamide slab gel containing SDS. As only small amounts of protein are involved, the SDS in the gel is sufficient to bind to the proteins as they migrate into the slab gel from the rod. Thus the second dimension is SDS-PAGE, with the proteins being separated by size.

What to do:From the PAGE menu, select 1-Dimensional PAGE.

Beware, once opened, never close the electrophoresis window from the control menu box (the square in the top left-hand corner of the window), otherwise you will not be able to perform any more electrophoresis. When you have finished a separation, click on Hide gel.

a) Perform an electrophoretic separation of the protein mixture using 1D SDS-PAGE.

(i) How many proteins appear to be present in the mixture? …………………………………………….(1 point)

(ii) Using the protein standards for reference, estimate the Mr of the subunits of each of the proteins in the mixture.

………………………………………………………………………………………………………………………………..(1 point)

(iii) Using the immunoblot (under PAGE select Immunoblot) identify which of the bands is protein 1.

………………………………………………………………………………………………………………………………..(1 point)

b) Now use 2D SDS-PAGE to analyse the simple protein mixture. (Choose Hide gel to leave the 1D SDS-PAGE screen; then select 2-Dimensional PAGE from the PAGE menu.)

(i) How many proteins appear to be present in the mixture now? ……………………………………..(1 point)

(ii) Explain what you would see if you only performed isoelectric focusing on this mixture.

………………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………(2 points)

(iii) What is the advantage of using 2D SDS-PAGE over the 1-dimensional technique?

………………………………………………………………………………………………………………………………………………..

…………………………………………………………………………………………………………………………………..(1 point)

c) Use the immunoblot facility to identify each of the proteins in the mixture, and complete the table. (This will involve selecting the proteins from this simple mixture in turn – see #5 on the previous page)

(3 points)

d) In some of the exercises which follow, you will need to use 2D SDS-PAGE to discover the effectiveness of various separation techniques. You will therefore probably find it helpful sketch the positions of the proteins on this diagram, and label which one is which.

(3 points)

e) (i) From the results of the 2D SDS-PAGE can you be certain that there are only three proteins present in this mixture?

………………………………………………………………………………………………………………………….(1 point)

………………………………………………………………………………………………………………………………………………….

………………………………………………………………………………………………………………………………………(1 point)

2)    Ion-exchange chromatography

Introduction: Ion-exchange chromatography separates proteins on the basis of charge using an ion-exchange resin. An ion-exchange resin consists of an insoluble matrix with charged groups covalently attached. A cation-exchange resin is negatively charged, and binds positively charged ions (cations). Similarly, a positively charged resin is called an anion-exchanger. An ion which binds weakly to the resin may be displaced, or exchanged, by an ion that binds more strongly. The degree to which a protein is retained by an ion-exchange column depends on the sign and magnitude of the protein’s net charge. The overall charge of a protein depends upon the number and type of ionisable amino acid side chain groups, and the pH of its surroundings. Each ionisable side chain group has a distinct pKa, that is the pH at which it is half dissociated. For each protein there will be a pH at which the overall number of negative charges equals the number of positive charges and so it has no net charge. This is its isoionic point (pI). If the pH is below the pI the protein, then the protein molecules are positively charged and will bind to a cation exchanger. If the pH is above the pI, then the protein is negatively charged, and so will bind to an anion exchange resin. A pH equal to the pI results in the protein molecule carrying no net charge and so it will not bind to either type of exchange resin. In selecting which type of exchange resin to use, it is important to consider the pH range over which the protein is stable (and therefore functionally active).

An ion-exchange resin is mixed with a suitable buffer, of an appropriate pH, to form a slurry. This is then poured into a chromatography column. The pH of the buffer will determine the charge on the proteins to be separated. The pH of the starting buffer should be at least one pH unit above or below the pI of the protein to be bound to the resin, to ensure adequate binding. It is also important to bear in mind the pH ranges of the ion-exchangers. A weak ion-exchanger is ionised over only a limited pH range (the term ‘weak’ does not refer to the strength of the binding of the ions to the resin). The resin in the column is washed with the starting buffer, then the protein mixture is applied. Proteins will bind or pass straight through the column, depending on their charge relative to that of the resin. Those that have been bound can be eluted by changing either the pH or the ionic strength of the eluting buffer. At low ionic strength there is minimal competition between the buffer ions and the proteins for charged groups on the ion-exchanger, and so the proteins bind strongly. As the ionic strength is increased the competition increases and so the interaction between the ion-exchanger and the proteins is reduced, causing the proteins to elute, regardless of the type of ion-exchanger used.

In this exercise you will investigate the binding of the proteins from the simple mixture, to two different ion-exchange resins, using both salt (ionic strength) and pH gradients.

What to do: Choose a pair of ion-exchange media to experiment with, either DEAE- and CM-cellulose, or Q- and S-Sepharose. (Tick the appropriate box).

As this is the first time you will have encountered a chromatographic technique in this program, take some time to investigate the menus and the options available, using the following notes to help you. It is worth doing, as many of the other separation techniques in Protein Purification are presented in a similar way

Select Ion exchange chromatography from the Separation menu. A menu pops up; choose one of the ion exchange media from the top list by clicking on it (you can change your mind by clicking on another from the list) and an elution method from the bottom list; when you are happy with your selection click on OK. Another dialog box appears; type in the pH of the equilibration buffer and click on OK. A third box appears in which you enter the values for the start and end of your chosen type of gradient (either salt or pH), then click on OK. A graph, or “chromatogram” will appear. This shows the amount of protein, as detected by UV-absorption, (left-hand y-axis) against the fraction number (x-axis), and also the gradient used (scale on right-hand y-axis).

There are a number of things you can do from this screen, (and this is the same for the other separation techniques which produce a similar type of display).

ར  You can perform an enzyme assay on the fractions, to discover the location of the protein of interest. From the Fractions menu select Assay enzyme activity. A graph of enzyme activity is superimposed on the chromatogram.

ར  You can perform 1D SDS-PAGE on selected fractions. From the PAGE menu select 1-Dimensional PAGE. A box appears to tell you how to select up to 15 fractions for electrophoretic analysis. When you have read this, click on OK to continue, or cancel if you have changed your mind. This is another useful way of finding your selected protein, and checking for contaminants in a chosen fraction.

ར  You can perform 2D SDS-PAGE on selected fractions. From the PAGE menu select 2-Dimensional PAGE. Again a box appears to tell you how to indicate the fraction to be analysed.

ར  If you want to select a certain group of fractions for use in the next step of your purification scheme, from the Fractions menu choose Pool fractions. A box appears, telling you how to use the mouse to select the fractions; you can cancel at this stage, or click on OK to continue. Slide the arrow to the first of the fractions, click the left-hand mouse button, point the arrow to the last of the fractions and click with the mouse again. Select carefully with the mouse, as you do not get the chance to undo your selection. A results box appears; click on OK to continue. (The results in this box are summarised in Progress report, available from the Help menu.)

ར  If you are unhappy with the results of this separation step, you can choose to Abandon this step and continue (available in the Quit menu). (NB You do not have this option if you have already pooled your fractions.).

ར  If you wish to abandon your entire separation scheme, choose Abandon scheme and start again from the Quit menu. If this option is not available, choose Abandon this step first, and then Abandon scheme on the next screen. You will be asked if you really want to do this (as it means that you lose all your data), if you choose No you will be returned to the screen you just left, if you choose Yes, the box appears saying Start from stored material?

a) Using your anion exchanger with salt elution, perform three experiments using an increasing salt gradient of 0-1 M, one at pH 5, another at pH 7 and the third at pH 8.

(i) Report your results using the table (enter fraction numbers in the form 27-38, and indicate the proteins present in each peak by their identification numbers):

(6 points)

(ii) At pH 5, what is the charge (positive or negative) on protein 1?……………………….

Protein 2?………………………….   Protein 3?…………………………… (1 point)

What is the charge of the column (the anion exchanger)?………………………………………………….(1 point)

Will any of the proteins bind to the column? If so, which ones?…………………………………………(1 point)

At pH 7, what is the charge on protein 1?…………….. Protein 2?…………….. Protein 3?…………..  (1 point)

Will any of the proteins bind to the column, and if so, which?…………………………………………….(1 point)

At pH 8, what is the charge on protein 1?…………….. Protein 2?…………….. Protein 3?…………… (1 point)

Will any of the proteins bind to the column, and if so, which?…………………………………………….(1 point)

(iii) At pH 7, which peak (peak 1 or peak 2) represents proteins which bound to the column? ……………

Which peak represents proteins which did NOT bind to the column?………………………………….(1 point)

b) How would your results differ if you had used a cation exchanger? Be specific! (1 point per pH)

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

c) (i) Use the anion exchanger to investigate the effects of changing the pH gradient on the separation of the protein mixture and record the results in this table. (6 points)

(ii) Comment on the results you observed. (3 points, one for each pH)

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

d) (i) Which protein can be separated in a single step using ion-exchange chromatography?

…………………… (1 point)

…………………………………………………………………………………………………………………………………………

………………………………………………………………………………………………………………………………………….

3)    Gel filtration

Introduction: Gel filtration separates proteins on the basis of differences in their size and shape. The technique uses a gel matrix consisting of porous beads of an inert, highly hydrated gel. The gel beads are packed into a glass or plastic column, and then equilibrated with a suitable buffer solution. The protein mixture is applied to the top of the column and then buffer is added to elute the proteins from the column. The eluate is collected at the base of the column as a series of fractions. As the proteins pass down the column they penetrate the pores of the gel beads to different extents, and so travel down the column at different rates. All proteins which exceed the maximum size of the pores will be unable to enter the beads. Therefore these proteins will only pass through the solution between the beads, and so elute from the column first, in the exclusion (or void) volume. All proteins smaller than the minimum size of the pores will equilibrate completely with the buffer inside and outside the gel beads, and so spend a proportion of their time inside the beads. These proteins will therefore move more slowly through the column and will be eluted last. These proteins elute in a volume very close to the bed (total) volume of the column. The pores in the beads are not all exactly identical in size, but span a narrow range of sizes. Proteins that have sizes very similar to the range of pore sizes will be excluded from some pores, whilst entering others. These proteins of intermediate size will therefore be partially excluded from the beads to an extent that depends on their size and shape. They will elute from the column in order of molecular mass, with the largest proteins eluting first and the smallest proteins last.

What to do: Use the various gel media available in this program to investigate the principles of gel filtration. There are three series of gel media to choose from, and each gel type is available with a range of pore sizes:

b Ultrogel is a registered trademark of Pharmacia-LKB

From the separation menu select Gel filtration. A menu pops up; choose one of the gel media from the list by clicking on it, you can change your mind by clicking on another from the list; when you are happy with your selection click on OK. A graph, or “chromatogram” will appear. This shows the amount of protein, as detected by UV-absorption, (y-axis) against the fraction number (x-axis).

a) Using the series of Ultrogel media examine the effect of increasing the pore size of the gel on the separation of the protein mixture.

(i) Complete this table with the fraction numbers occupied by each peak (eg. 32-45): 3 points

(ii) What conclusion can you make regarding the time of elution of a particular protein, as the pore size of the gel is increased? (2 points)

…………………………………………………………………………………………………………………………………………

(iii) Using this technique is it possible to separate the following pairs of proteins from each other? Complete the table below using yes if a pair can be separated and no if they cannot. (3 points)

(iv) Reconcile the results observed here with those you recorded in the table for exercise 1) c).

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………..(2 points)

b) Perform a separation of the protein mixture using Sephadex G-50.

(i) In which fractions are the proteins eluting? ……………………………………………………… (1 point)

(ii) What do you think has happened to the proteins in the first peak?

…………………………………………………………………………………………………………………………(1 point)

(iii) Suggest how the resolution of this separation could be improved, using the Sephadex gel series.

……………………………………………………………………………………………………………………………………

……………………………………………………………………………………………………………………(2 points)

c) Perform a separation of the protein mixture using Bio-Gel P-300.

(i) What results do you observe? …………………………………………………………………………..(1 point)

(ii) Explain what has happened to the proteins during this separation.

…………………………………………………………………………………………………………………………………………

………………………………………………………………………………………………………………………….(1 point)

(iii) Under what circumstances would Bio-Gel P-300 be the most suitable gel medium to separate a group of proteins?

………………………………………………………………………………………………………………………….(1 point)

The point values add up to 60 points; your score will be divided by 3 to give a possible total of 20 points.

4)    Isoelectric focusing(IEF)

Introduction:Isoelectric focusing (IEF) is a method for separating molecules which differ in their charge characteristics. For IEF of proteins, the protein mixture is subjected to an electric field in an inert support medium in which a stable pH gradient has previously been generated. The inert support can be either agarose or polyacrylamide. The pH gradient is formed in this by including a mixture of low molecular mass “carrier ampholytes”. The anode (positive electrode) region is at a lower pH than the cathode (negative electrode). The pH range is chosen such that the proteins to be separated have their isoelectric points within this range. A protein which is in a pH region below its pI will be positively charged, and so will migrate towards the cathode. However, as it migrates, so the pH that the protein experiences will decrease until the protein reaches a pH which is equal to its pI. At this point it has no net charge and so migration ceases. Should the protein overshoot this point, it will enter a region of pH above its pI and so become negatively charged. It will then reverse its direction of migration and now migrate towards the anode. Therefore proteins become focused into sharp stationary bands, with each protein positioned at a point in the pH gradient corresponding to its pI. The technique is capable of extremely high resolution with proteins differing by only a single charge being resolved. It is important to avoid molecular sieving effects so that the protein separation occurs solely on the basis of charge, so the chosen support medium has pores larger than the size of the proteins being separated. IEF is mainly an analytical tool, but can be used to prepare very small amounts of pure protein. In preparative IEF, if the separation has been performed in a slab gel or in a tray of gel beads, then the bands can be cut from the gel and the proteins eluted using a buffer solution.

What to do: Select Preparative isoelectric focusingfrom the Separation menu. A box appears in which you enter the pH values for the start and end of the pH gradient, then click on OK.

In this exercise you are going to explore the effects of pH range on the separation of proteins by isoelectric focusing. Conduct your own experiments and report briefly on your findings and conclusions . Use the following suggestions to help you:

1.    taking a wide range around the pI of the chosen protein

2.    using a narrow range around the pI of the chosen protein

3.    using a range which excludes the pI of the chosen protein

……………………………………………………………………………………………………………………

……………………………………………………………………………………………………………………

……………………………………………………………………………………………………………………

……………………………………………………………………………………………………………………

……………………………………………………………………………………………………………………

……………………………………………………………………………………………………………………

……………………………………………………………………………………………………………………

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5)    Heat treatment

Introduction: Protein purification procedures are usually carried out at low temperature (0-4C) since most proteins are stable at low temperatures. As the temperature increases from 0C to 37-40C their stability decreases significantly. Above 40C or so, most proteins become increasingly unstable and denature. At neutral pH, denatured proteins usually precipitate. Individual proteins differ in their heat sensitivity and so this can be used for purification purposes. The temperature stability of the desired protein is determined by trial experiment, for example by following enzyme activity as the protein mixture is incubated at different temperatures, for a set period of time. The minimum temperature at which gross inactivation occurs is noted. Once this temperature is known, less stable proteins can be preferentially inactivated by incubating the crude protein mixture at a temperature 5-10C below this value for 15 to 30 minutes. Since denaturation of all cell proteins occurs to some extent at all temperatures, and only increases with increasing temperature, the total activity of the desired enzyme usually falls to some extent after heat treatment. However, it may be a useful early step for the purification of rather more heat-stable proteins.

What to do: Use Heat denaturation (available in the Separation menu) to answer these questions:

a) Which of the proteins in this simple mixture is suitable for purification using heat treatment?

………………

b) Explain your choice of protein for question a).

…………………………………………………………………………………………………………………………………………..

c) For how long does the protein mixture need to be held at the following temperatures before all of the contaminating proteins are precipitated?

(i) 60C ……………………………………..

(ii) 50C …………………………………….

d) Why, in practice would it not be a good idea to perform a one step protein purification in this way?

…………………………………………………………………………………………………………………………………………

6)    Ammonium sulphate fractionation

Introduction: The solubility of proteins varies according to the ionic strength, and hence according to the salt concentration, of the solution. Two distinct effects are observed. At low concentrations of salt, the solubility of the protein increases with salt concentration. This phenomenon is called “salting in”. However, as the salt concentration (ionic strength) is increased still further, the solubility of the protein begins to decrease. At sufficiently high ionic strength the protein will be almost completely precipitated from solution – an effect called “salting out”. Since proteins differ markedly in their solubilities at high ionic strength, salting-out is a very useful procedure to assist in the purification of a given protein. The commonly used salt is ammonium sulphate, as it is very water soluble and has no adverse effects upon enzyme activity. It is generally used as a saturated aqueous solution which is diluted to the required concentration, expressed as a percentage concentration of the saturated solution (a 100% solution). Before carrying out a bulk separation, a test is performed to discover the salt concentrations to use. In this preliminary test the concentration of ammonium sulphate is increased stepwise, and the precipitated protein is recovered at each stage. Each protein precipitate is dissolved individually in fresh buffer and assayed for both total protein content and the amount of the desired protein (in this case an enzyme, measured by its activity). The aim is to find an ammonium sulphate concentration which will precipitate the maximum proportion of undesired protein, whilst leaving most of the desired enzyme still in solution. The precipitated protein is then removed by centrifugation and then the ammonium sulphate concentration of the remaining solution is increased to a value that will precipitate most of the enzyme whilst leaving the maximum amount of protein contaminants still in solution. The precipitated enzyme of interest is recovered by centrifugation and dissolved in fresh buffer for the next stage of purification. This technique of ammonium sulphate fractionation is extremely useful to quickly remove large amounts of contaminant proteins, as a first step in many purification schemes. It is also often employed at later stages of purification; to concentrate protein from dilute solution following procedures such as gel filtration.

What to do: Use Ammonium sulphate fractionation (available in the Separation menu) to discover a suitable protocol for protein purification using this technique. In order to carry out preliminary tests on the protein mixture, in the first box type the chosen concentration of ammonium sulphate, click on OK, a results box appears. If you wish to go back and try another concentration click on cancel in this results box, but if you wish to continue to another step in the purification, choose whether you want to use the precipitate or the supernatant by clicking on the appropriate word, then click on OK.

a) What is the maximum concentration (to the nearest whole number) of ammonium sulphate that can be added to this protein mixture without precipitating any of protein 2?

………………………………………………………………….

b) What is the minimum concentration (to the nearest whole number) of ammonium sulphate that can be added to the protein mixture to precipitate all of protein 2?

………………………………………………………………….

c) Is it possible to obtain a pure sample of any of the proteins in this mixture using ammonium sulphate precipitation alone?

………………………………………………………………….

7)    Hydrophobic interaction chromatography

Introduction: This technique separates proteins on the basis of their binding to and elution from a hydrophobic matrix, usually octyl- or phenyl-agarose. Binding of the proteins is often carried out at a high salt concentration to favour hydrophobic interactions. Some proteins may precipitate at this high ionic strength and so need to be removed by centrifugation prior to loading the protein mixture onto the column. Selective elution of bound proteins is then carried out by applying a decreasing salt gradient.

What to do: Explore the effects of changing the starting and final salt concentrations on the separation of the three proteins using Hydrophobic interaction chromatography(select it from the Separation menu).Choose either Phenyl- or Octyl Sepharose (use the same one for the whole of this exercise) and click on OK. Enter the values the Start and End of the salt gradient in the boxes provided, and click on OK.

I am using: (check one)

a) What are the two effects of increasing the initial salt concentration on the elution profile?

(i)………………………………………………………………………………………………………………………………………

(ii)……………………………………………………………………………………………………………………………………..

b) What are the effects of increasing the final salt concentration on the elution profile?

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

c) Which protein can be separated in a single step using this technique, without losing any activity?

…………………………………………………………………………………………………………………………………………

d) Investigate a series of fractions through the second peak eluted using 1D-SDS PAGE, what do you notice?

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

8)    Affinity chromatography

Introduction: Affinity chromatography relies on the preparation of a matrix to which the protein of interest, and preferably only this protein, will bind reversibly. The matrix is usually beaded agarose, polyacrylamide or cross-linked dextran, to which a ligand has been covalently attached. The chemical nature of the ligand is determined by the known biological specificity of the protein to be purified. In the case of an enzyme, the ligand chosen would probably be a substrate or a reversible inhibitor or activator. If it is not possible to use a ligand that is absolutely specific to the molecule of interest, it is often possible to use a group-specific ligand. The matrix bearing the ligand is packed into a column, in a buffer that will be optimal for the protein-ligand binding. For example, if the ligand is an enzyme substrate, then the buffer must contain any co-factors that are required for binding. The buffer usually has a fairly high ionic strength, to minimise non-specific binding of other proteins to the ligand. The sample is applied at the top of the column and washed through the matrix. Ideally, only the protein of interest should bind. It can then be eluted specifically by the addition of a relatively high concentration of substrate or competitive inhibitor, or, failing this, by changing the pH and/or the ionic strength to disrupt the enzyme-ligand interaction. An alternative protocol can be used if an antibody, specific to the protein of interest, is available. This procedure is applicable to all proteins irrespective of their functional activities. The antibody is covalently coupled to a suitable matrix filling the column. Only the required protein will bind to the antibody and can then be eluted by procedures which weaken the antibody-antigen interaction. Affinity chromatography is a potentially powerful technique, but it can only be used when the functional activity of the required protein is known and a suitable ligand is available, or when a suitable antibody to the protein has already been obtained. Unfortunately, in many cases neither condition is satisfied, and so other protein purification methods have to be relied upon.

What to do: Use Affinity chromatography (from the Separation menu) to discover suitable purification methods for the proteins in this mixture. You will need to experiment, as you have no way of knowing the affinity of the various monoclonal antibodies available for each protein. Select a ligand from the left hand list, and choose what you want to elute the proteins with, from the list on the right, then click on OK.

a) For protein 2, discover which monoclonal antibody and elution system gives the best separation and yield.

…………………………………………………………………………………………………………………………………………

b) For protein 1, explain the results you observe when using monoclonal antibody MC01C.

…………………………………………………………………………………………………………………………………………

c) For protein 3, compare the results you observe when using monoclonal antibody MC03C with those for the polyclonal IgG, and explain any differences you observe.

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

d) (i) Which protein has a competitive inhibitor that can be immobilised? ……………………………………………………..

(ii) Which is the most efficient method for performing a separation using this competitive inhibitor? Explain.

…………………………………………………………………………………………………………………………………………

…………………………………………………………………………………………………………………………………………

(iii) How does the yield of protein for the method in d) (ii) compare to the best yield available for this same protein using other methods of affinity chromatography?

…………………………………………………………………………………………………………………………………………

9)    And finally………..

Suggest a two step purification process capable of separating all three of the proteins from each other. There is no single correct answer to this, we will discuss the various options available in the group discussion.

…………………………………………………………………………………………………………………………………………

PART II.  Purifying a protein from a complex mixture

WRITTEN WORK FOR PROTEIN PURIFICATION LAB

Answers to the following questions are due before you do the computer exercise.  All but one can be answered by using the previous sections.

Questions:

1.  Since proteins are quite fragile (labile) molecules, all purification steps should take into consideration the following conditions

1.

2.

3.

2.  Early in purification, two low resolving steps often used are

1.

2.

while a low capacity but high resolution step often used late in purification

is______________________________.

3.  Traditionally, one unit of enzyme activity is defined as

4.  As the enzyme is purified from a mixture of proteins we measure its purity by determining the number of units of enzyme activity per milligram of protein, this measure is the term _____________ _____________of the enzyme.

5.  “Fold” purification compares the _____________________of a fraction to that of the original mix.

6.  Enzyme yield is defined as:

7.  We will consider our purification is probably complete when the apparently pure protein yields one spot when tested by _________________________.

8.  When choosing a gel type for optimal fractionation of complex proteins the gel pore size is such that the desired protein is _____________________________________________.

9.  The ph at which a protein has no net charge is its ________________________________.

At this point it (will, will not) bind to an ion exchange resin.  Below this pH it will assume a __________ charge and bind to a (cation, anion) exchanger.

10.  DEAE cellulose or CM cellulose are effective only in the pH range ___to____.  The starting buffer should be of reasonably __________ ionic strength the affinity of proteins for the ion exchange resins (increases, decreases) as ionic strength (increases, decreases).

11.  The O.D.280  is monitored in each of the eluted fractions because

__________________________________________________________________________

___________________________________________________________________________.

Turn in the “Record of Purification” for two enzymes as well as the 2-D Page gel with the enzymes of interest circled. Be sure to identify them by number.  Remember it is possible that one of your enzymes is a dimer or that the isoelectric point of your enzyme is beyond the range of pH values displayed on the gel.

The report should concisely summarise your results (yield, purity and so on) for each stage of the purification, and your conclusions. You should say why you have chosen to use particular separation techniques for a given protein. The report should also include any relevant details regarding the characterisation of the proteins in question, for example estimated Mr of the subunit. Your report should conclude with your recommendations for the way in which the project should proceed, in terms of the optimum separation technique for a given protein. You do not have to report everything you have done, but if you discover that a particular separation technique is entirely unsuitable for a given protein, you could mention this, to save time for other researchers in the future.

It is strongly recommended that you keep careful notes of everything you do while you are carrying out your investigations; remember that the program does not record everything for you. However, you should not include all that you write down in your final report. Your notes will form the equivalent of a lab note book; you would not publish the entire contents of such a book as a scientific paper!

You will be assigned2 proteins to purify; you must clearly write the number of the protein to be purified at the start of the relevant section of your report. Failure to do this will result in your assignment not being marked.

You will purifyeach of your 2 proteins from a complex protein mixture in a crude mucosal extract. In each case the protein you must purify has not been isolated before. You are aware of its enzymic activity, so you can detect its presence, but there are no specific antibodies available. You have to discover the most efficient and cost effective separation method for each of the proteins assigned to you, so you need to carry out several investigations using different combinations of separation techniques to discover the optimal method. You are aiming to get a pure sample of your protein, with a high yield. The budget allocated to your project is restricted, so you must pay careful attention to the cost of each step in the purification. Your research director will monitor the progress of your project, both in terms of time and financial costs, and will intervene if it appears that you cannot meet the time and budget targets.

The proteins I have been assigned to purify from the crude extract are:

…………………………&…………………………..

What to do: BeginProtein Purification. On the title page, click on Start and select Choose a mixture from the drop-down menu. Choose the Default Mixture and type in the number of your assigned protein.

A common strategy is to run a separation, pool your samples, and then run a 2D electrophoresis experiment to evaluate your results.  A quirk of this program is that you then have to select “hide gel” from the menu before you can continue.

## Research Paper Writing- Finance

HILTON HOTEL

PURPOSE

The purpose of this project is two-fold:

1) to become sufficient in basic Excel functionality, including formulas and formatting

2) to analyze the operations of a hospitality company

In addition this project will address the following learning objectives required of this course.

-Inquiry and Critical Thinking: Graduates are able to identify problems, articulate questions,  and use various forms of research and reasoning to guide the collection, analysis, and use of  information related to those problems.

-Communication: Graduates are able to write and speak effectively to both general and specialized audiences, create effective visuals that support written or spoken communication, and use electronic media common to one’s field or profession.

Hospitality College Learning Objectives:

-Communicate effectively in written, spoken, visual and digital modes to different audiences  (e.g. industry leaders, employees, employers, faculty and peers).

-Manage all forms of capital (e.g., human, financial) in an ethical and sustainable way.

-Analyze financial, marketing, and operational results and outcomes for hospitality operations.

-Demonstrate effective management techniques in hospitality operations (hotel, F&B, gaming, meetings, events, etc.).

You will prepare the income statement and balance sheet for 2 calendar years (2018-2019), conduct a horizontal and vertical analysis, and calculate ratios of the company you choose.  As a group you will analyze these financials statements and ratios to evaluate your company performance between the 2 years.

This project is intended to integrate the material learned in class and specifically Chapters 2, 3, and 5.  This project will give students an application example that is very common in industry.  This project is typically the first steps hospitality management takes in not only analyzing their financial performance and making adjustments if necessary, but also preparing to conduct their forecast and budgets, which we cover in Chapter 9 and 10 (and which you will do in Project 2).

Submissions are expected to resemble those presented to an upper level manager in a hospitality operation and will give students an opportunity to apply written communication skills.

This project is to be completed using a publicly traded hospitality company’s financial statements (10-Ks).  Each group must choose a different hospitality company.

Vertical and Horizontal Analysis

You are to start by building in Excel the year end 2019 and 2018 Income Statement and Balance Sheet for your hospitality firm based off the company’s 10-K (If there is a 10-KA must use that).  All statements should have account names first than the 2019 numbers in the second column and 2018 in the third column.  No other years will be accepted without prior approval.  If your company does not do a calendar year end (or within 1 month of the year end) you must email me what their year-end is and get approval.

The Income Statement must have subtotals for Total Revenue, Total Operating Expenses, Operating Income, Income Before Taxes, and Net Income at a minimum. You may have more subtotals based on the company’s financial statements but you MUST have these 5 at a minimum.  You must include all subtotals off the 10-K in addition to these stated.  All subtotals must be formulas and not hardcoded numbers.  If you believe your company financials do not have these subtotals you must check with the professor for a variation to this requirement.  Anything turned in without these will be penalized points. If you do not know what a subtotal should include do not ignore it, come to office hours to ask.

Only go to Net Income or Net Loss.  If your company has multiple Net Income/(Loss) lines due to subsidiary companies and such, go all the way to the last one right before earning per share.  Do not include any Earnings Per Share information or additional information below that.

The Balance Sheet must have subtotals and totals for Current Assets, Long-term Assets, Other Assets, Total Assets, Current Liabilities, Long-term Liabilities, Total Liabilities, Total Stockholders’ Equity, and Total Liabilities and Stockholders’ Equity at a minimum. More subtotals may be added based on the company’s financial statements, but you MUST have these 9 even if the company does not show these subtotals.  You must include all subtotals off the 10-K. All subtotals and totals must be formulas and not hardcoded numbers.

You are to do a horizontal and vertical analysis for each statement.  You are to do all vertical and horizontal analysis possible given that you only have 2 years of data.  Do not do any more years.

Ratios

Calculate the following ratios for both years:

Current Ratio (coverage ratio)

Working Capital (show to nearest dollar, ie. \$10,000)

Debt-to-Equity (coverage ratio)

Asset Turnover (turnover ratio)

Profit Margin (percentage ratio)

Return on Assets (percentage ratio)

Return on Equity (percentage ratio)

Earnings Per Share (Per unit basis – show in dollars and cents, ie. \$1.25)

If your company is not publicly traded, you must check with the professor to get an alternative ratio to calculate.  You cannot just ignore this ratio.

Solvency Ratio (coverage ratio)

Number of Times Interest Earned (coverage ratio) – If you company does not have interest expense you must see the professor to validate and be given an alternative ratio to calculate.  If you calculate this and have no interest and did not check you will receive no credit on this ratio.

1. For any formulas that use Net Income, you must use your company’s bottom Net Income (the last one).  Using a different Net Income line will cause you to get no points.
1. Some of these ratios may require you to get additional information that is not on the financial statements.  Do not ignore those ratios, make sure to get the additional information.
1. All ratios must be linked to the Income Statement and/or Balance Sheet and the only numbers allowed to be hardcoded are those that do not appear on the financial statements.

Write-up

Based on the horizontal and vertical analysis and the ratios you calculated, as a group select 5 accounts per statement that as management of this company you would want to analyze.  You must select accounts not subtotals or totals.  You should have a write-up on 10 accounts (5 from the Income Statement and 5 from the Balance Sheet).  Each account must answer the following 2 questions (See Criteria section for an example).  You cannot select Accounts Receivable (or an equivalent account) as an account to analyze since the example given below is on that account.

For each account you must provide a short explanation (1-2 sentences) of why you choose to analyze this account.  Your reasoning for account selection should be based on class discussions, what you have learned in class, and the results of your horizontal and vertical analysis or ratio calculations.  Specifically discuss what on the horizontal and vertical analysis or ratios calculations made you choose this account and this includes stating the exact numbers that caused you to select these accounts.  If you choose an account that had minimal change based on your analysis you must support why you choose to look into an account that did not have a big change.

After discussing why you selected the account you must explain what caused the change in the account.  You must get this information from the 10-K directly and cite the page number you got your information from.  If you cannot find a valid explanation for the change you must select another account (although most accounts have a reason but it may not be explicitly stated).  If you do not understand the change, please see the professor for assistance.  Do not guess.  Make sure you include the correct reason.  For instance if the account went down (like in the example below for accounts receivable) you cannot state that the allowance for doubtful accounts, which is a contra asset account went down.  This account going down would cause Accounts Receivable to go up. Also make sure to address an issue that was a main reason for the change.  For instance, if the account changed \$10 million, do not include an explanation for \$1 million of the change when there is also information on \$4 million of the change.

Team Member Evaluation

If you want (not required though) you may do a team member evaluation and turn it in on the same day you turn your project in.  Just like working in teams in the real world, sometimes certain members of a group do not do their share of work even though everyone gets the same credit.  After reviewing the evaluations if the majority of team members give significantly lower scores and is supported with comments and proof such as unanswered emails, that team member’s score will be lowered and if the member did no work they will receive a “0”.  The evaluations along with all supporting proof should be turned in on the same date and can be turned in during office hours instead of in the classroom. Evaluations turned in with no proof will not be accepted. The instructor may request a meeting with you about your evaluation before adjusting teammates grades.

CRITERIA

Due Date

The project is due on Wednesday, October 14, 2020.  Projects received after this date will be penalized 10 points (out of 100) each 24-hour period they are late (beginning at the start of class).  This includes not submitting both parts online by the due date.  You can earn extra credit by turning the project in at the beginning of class or before Wednesday, October 7, 2020.  All projects turned in for extra credit will earn 1% point additional on your final class grade.

Projects must be submitted online (regular due date and extra credit date).  No projects will be accepted for extra credit after the start of class on the due date, including even one minute late.  Any projects not completed are not eligible for extra credit including not having all pages or parts attached.

It is imperative you turn in professional work and not leave this until the last minute.  I will not accept unprofessional work just as your manager would not.  Unprofessional work is not college level, it’s extremely hard to grade, and the scores are extremely low.  If I determine your work is unprofessional, such as no formatting on the statements, multiple pages per statement, papers not straight or unreadable, etc. you will incur a 15 point penalty and be informed via email that you then have ONLY 24 hours to fix and resubmit.

Requirements

All vertical and horizontal analysis and ratios must be completed using Excel. All write-ups must be completed in Word.  Handwritten assignments or portion thereof are unacceptable.

You must have a vertical and horizontal analysis for the Income Statement and Balance Sheet. All statements (4 in total) and ratios should be printed “portrait” style and be on one sheet of paper each.  If you cannot fit them on one sheet of paper you must check with the professor beforehand to get approval for multiple pages or points will be taken off.  To get approval please bring in your copy and a copy of the 10-K statement. (Note: in case of remote learning, you will not need to submit hardcopy).

You must do this in Excel and I will be reviewing all electronic submissions to confirm you did all required formulas in Excel.  Not doing this project in Excel, not submitting this online, or not using the Excel formulas will result in no credit for this part and hence a 60% point penalty on the entire project grade.

On the horizontal and vertical analysis, all dollar figures should be presented exactly like the 10-K and all percentage columns should be shown as percentages with 1 decimal (i.e. 7.2%).  Make sure that your all statements are formatted with dollar signs, commas, borders, underlining, etc. and are presentable.  Also make sure to note what value your company’s dollar values are in, for instance thousands or millions.

On the ratios, dollar figures should be rounded to the nearest dollar or cents depending on ratio, coverage and turnover ratios should be presented to 2 decimals (i.e. 1.12 times or 1.12 to 1), and percentage ratios should be shown as percentages with 2 decimal (i.e. 7.25%).  Make sure that all ratios are formatted correctly and are presentable.

The write-ups must also be portrait style and must be double spaced in 12-point font.

You must submit your Excel submission and your Word submission to WebCampus and check your plagiarism score. Both are due before the due date and time online in addition to being turned in in class.

Spelling and grammar count.  Remember you are presenting to upper management and the report should be written with that in mind.  Run spell check in both Excel and Word. It is suggested you see the Writing Center if your writing skills are lacking.  Points will be deducted for papers that are hard to read and if they are too hard to read it will be a “0”.

You must submit hard copies (or word file, if the class is remote) of each part of the project and you must present your work as follows in order to receive credit:

Individual Excel Analyses and Ratios

Income Statement horizontal analysis

Income Statement vertical analysis

Balance Sheet horizontal analysis

Balance Sheet vertical analysis

Ratio calculations

Print out of the Income Statement from the company’s 10-K for all years used

Print out of the Balance Sheet from the company’s 10-K for all years used

Group Write-ups

Cover Page with all students’ names listed, the name of your company, and the originality score your received when you uploaded the write-ups.  The originality score can be handwritten.

Income Statement write-up

Balance Sheet write-up

Print out all pages from the 10-K that you cite in your write-up.  Put them in order of how you cite and highlight the area you cited.  Also make sure each is numbered and in order.

1. This should be presented as a “unified” group project.  This means that all sheets must be in the same font, format, etc.  Do not just copy and paste each group member’s part into the document and submit without making sure it is consistent in format, tone, wording, etc.  Remember this is a group project so you should not use “I” and instead use “we”.

Staple all of your analysis together.  No binders or paperclips are allowed.

The grading rubric below should be carefully reviewed and followed.

The write-up should be no longer than ¼ page for each account and really only needs to be 2-3 sentences to address the questions sufficiently.  Here is an example of a write-up that would get full credit.

As management in Boyd Gaming, we choose to look into Accounts Receivable, net.  We choose this account because it decreased \$38.33 million and 58.5% from 2013 to 2014.  This account also decreased from 1.1% of total assets in 2013 to 0.6% of total assets in 2014.  This account decreased mainly due to the allowance for doubtful accounts, which lowers the amount of accounts receivable, decreasing \$19.8 million due to the company deconsolidating Borgata in Atlantic City on September 30, 2014, which caused Boyd Gaming to no longer record those assets, liabilities, and non-controlling interests in the property (Pg. 65).   This one decrease accounts for over half the change, 51.7%.

You will notice both the vertical and horizontal reasons were given for selecting the account.  That is not required.  You can select based only on 1 of those (or the ratio calculations) if you want, but I wanted to give you both scenarios so you can see how they should be written.

This is a high-level guide to make sure the project is completed and follows all the requirements.  This has been added since I have found that most points off are because students do not read all the instructions and make up their own “rules”. This is just a guide, so you don’t miss any minimum requirements.

These items are points that come right off the starting score of 100%:

These are the maximum points for each required part:

## Research Paper Writing-Lab Report

EET 1198                                                                                            NAME ____________________

LAB1- INTRODUCTION                                                                 DATE __________

1. A logic probe will be used to check the +5V output on the trainer. Connect, using short wires, the logic probe power clips to the +5V (red) and GND (black) outputs. If the +5V is working the logic probe display will read “F”.
• Go to eLearn and use the link under “Student Resources” to find the list of integrate circuits (ICs) that we may use in the lab. In this part of the lab you will test a 2-input AND gate. In order to do this a 74LS________ chip will be used. Look up the corresponding pin-out diagram by selecting “More Info” and determine which pins inside the IC should be used to test a single 2-input AND gate. Based on the circuit design that was done in lectureyou can test one of the gates inside that IC using pins ___, ___, & ___.
• For the 2 binary inputs we willuse two of the switches found on the trainer. For the outputwe will use LED7 on the trainer. LED7 will indicate 1 (on) & 0 (off).

Wire the inputs and output to the IC, use the switches to provide the inputs in the table, and mark the corresponding output as 0 (LED off) or 1 (LED on). Use your logic probe to confirm the high/low signals coming from the switches to the IC input pins. Show the working circuit to the instructor and get it signed off.

Instructor check _________

• In this part of the lab you will test an OR gate.

In order to do this a 74LS_____ chip will be used. To test one of the gates inside that IC requires using pins___, ___, & ___.

Build the circuit with the new IC, provide the given inputs from the table and record the corresponding outputs in the table. Show the working circuit to the instructor and get it signed off.

Instructor check _________

• Re-build each circuit in the MultiSim software. Show your working circuits to the instructor and get it signed off. Then print and turn in with your lab.

Instructor check _________

## Research Paper Writing- Lab Report

EET 1198                                                                                            NAME ____________________

ACTIVITY2                                                                                       DATE __________

1. Build the given circuit in Multisim. Connect each output (W, X, and Y) to a current limiting resistor and an LED to indicate its “ON”/”OFF” status. Make each switch so that it is controlled by different keys on the keyboard. The keys you choose should be in some logical order or location on the keyboard so that it is simple to toggle the switches on and off for all combinations. Test each input in the provided truth table, and record the corresponding outputs in the table. Show the working simulation to the instructor.

Teacher Check________

• Determine the correct IC, find the pinout diagram, and make a list of what each pin on the IC will be connected to below. Then, breadboard the circuit using the switches on the trainer for inputs and running the W, X, and Y outputs each to a current limiting resistor and an LED on the trainer. Verify your output for each input combination in the given truth table. Show the working circuit to the instructor.

Teacher Check________

Pin 1-

Pin 2-

Pin 3-

Pin 4-

Pin 5-

Pin 6-

Pin 7-

Pin 8-

Pin 9-

Pin 10-

Pin 11-

Pin 12-

Pin 13-

Pin 14-

## Research Paper Writing-Math

Medgar Evers College, CUNY Department of Mathematics

Math 136 Intermediate Algebra/Trigonometry                                       Class QUIZ # 2

Name: _________                          Section: ______ID:  _

1. Vocabulary Quiz. Match by completing the blanks with corresponding equivalent meanings below represented by its letters.
1.  Commutative Property
2.  Evaluate
3. _____Function
4.  Slope – Intercept Form
5. _____Simplify
6.  Distributive Property
7. _____ Standard equation of a line
8.  Point – Slope Form
9. _____ y-intercept
10.  Variable
11. _____ Solve
12.  Rate of Change
1. Y=mx + b
2. A symbol or combination of symbols representing a value orrelation
3. The property that associates two quantities in any definite order
4. A relation between two sets involving independent and dependent variables
5. The ratio of the change in Y and the change in X (m)
6. To determine or calculate the numerical value of an expression
7. To break down or make less complicated orcomplex
8. To work out the answer or solution to an equation
9. Intersection of y-axis and a line graph
10. Ax + By = C
11. Y – y1 = m(x – x1)
• Find the slope of the lines.

(a)

(b)

• Sketch the graph of theline with the intercepts shown below.

a) x-intercept = -1,y-intercept =5

• Evaluate the function f(x) = 3   at
• f(-2)
• f(2) + 3 .
• f(3g)
• Graph and find solution to the system of equations

2x + 3y = 6

X – 2y = 4

• Write the equation of  line in standard form perpendicular to a line 5x + 2y = 6 and passing through a point  (-2, 0)

## Research Paper Writing- Strategic Management

Strategic Management     Assignment A

Unless otherwise stated, answer in complete sentences, and be sure to use correct English, spelling and grammar. Sources must be cited in APA format. Your response should be four (4) double-spaced pages; refer to the “Assignment Format” page located on the Course Home page for specific format requirements.

You will soon find yourself fulfilling roles in organizations where the ability to think strategically about issues will make you much more valuable to employers. For example, your boss may ask you to present information on a new strategy to gain a competitive advantage in the marketplace to the board of directors.

Part A: Strategic Management

Part A refers to the material in Lesson 1 of this course. Using logical, clear writing, do the following:

1. Describe strategy and the strategic management process.
2. Define competitive advantage and describe the two approaches used to estimate a firm’s competitive advantages.
3. Explain why it is important to understand a firm’s strategy.

Part B: External Analysis

Part B refers to the material in Lesson 2 of this course. Using logical, clear writing, do the following:

1. Describe an external analysis.
2. Analyze the two levels of the environment.
3. Identify and define the three elements of the S-C-P model.

Part C: Internal Analysis

Part C refers to the material in Lesson 3 of this course. Using logical, clear writing, do the following:

1. Describe an internal analysis.
2. Explain resources and capabilities.
3. Describe the VRIO framework.

Part D refers to the material in Lesson 4 of this course. Using logical, clear writing, do the following:

1. Differentiate between business strategies and corporate strategies and define the nature of a cost-leadership strategy.
2. Identify six sources of cost advantages for firms.
3. Identify the most appropriate organizational structure for a firm pursuing a cost-leadership strategy.

Assignment B

Unless otherwise stated, answer in complete sentences, and be sure to use correct English, spelling and grammar. Sources must be cited in APA format. Your response should be four (4) double-spaced pages; refer to the “Assignment Format” page located on the Course Home page for specific format requirements.

Making strategic decisions are part of the territory when it comes to the job of an analyst. You’ll need to decide what to do in a given circumstance and then answer questions to explain your decisions. Let’s say you are a Strategic Analyst for a Fortune 500 Company, and your CEO has asked you how to position a specific business and product line to gain a competitive advantage in a single market. This scenario is the foundation of lessons 5, 6, 7, 8, covered in the textbook.

Part A: Product Differentiation

Part A refers to the material discussed in Lesson 5 of this course. Using logical, clear writing, do the following:

1. Define product differentiation and discuss the role that customer perceptions play in product differentiation.
2. Identify the three broad categories of product differentiation and two bases of differentiation under each category.
3. Explain the relationship between product differentiation and managerial creativity.

Part B: Flexibility and Real Options

Part B refers to the material discussed in Lesson 6 of this course. Using logical, clear writing, answer the following:

1. What is strategic flexibility? Why is it thought of as a third generic business-level strategy?
2. What are strategic options?
3. What are real options?

Part C: Collusion

Part C refers to the material discussed in Lesson 7 of this course. Using logical, clear writing, answer the following:

1. What is collusion?
2. What are the two types of collusion and how are they different?
3. How does signaling relate to collusion?

Part D: Vertical Integration

Part D refers to the material discussed in Lesson 8 of this course. Using logical, clear writing, do the following:

1. Define vertical integration and differentiate between forward vertical integration and backward vertical integration.
2. Identify the three fundamental explanations of how vertical integration can create value and discuss how value is created under each.
3. Identify three reasons a firm may be able to create value through vertical integration when most of its competitors are not able to create value through vertical integration.

## Research Paper Writing- Fixed Income and credit analysis

Part 1: (20 Marks) A. Calculate the dirty price, clean price, modified duration and modified convexity of the Government bonds as at the end of January 2020 and the end of July 2020. Discuss your results. (7 marks) B. Calculate the holding period return in each bond. Discuss your results. (5 marks) C. Calculate the modified duration and modified convexity for an equally-weighted portfolio of the four bonds (25% weight for each bond) at both dates (that is, both at the end of January 2020 and the end of July 2020). Estimate the holding period return for the portfolio over the six months between the two dates. Report on your findings. Compare and contrast the return and volatility of the portfolio, and the separate bonds at both dates. Discuss your results. (8 marks) Part 2: (20 Marks) A. Construct and present a yield curve, spot curve and forward curve as at the end of January 2020 and the end of July 2020. Consider the government bond information on the RBA website. Your spot curve and forward curve estimation should go out no more than five years. Present and discuss your findings. (8 marks) B. Review the predictive ability of the yield, spot and forward curves with the comprehensive reference to the relevant academic literature. Discuss the curves that you have estimated in Part 2 (A) regarding this literature. Consider the current COVID-19 pandemic issue, does the January 2020 forward curve appear to predict the six-month spot rates in July 2020? Comment. (12 marks)

## Research Paper Writing- Accounting

PROJECT ASSIGNMENT

ASSIGNED INVESTMENTS:Pull up the finance.yahoo.com web site on your PC. (2) Find Vanguard 500 Index Investor Fund (ticker symbol: VFINX).(3) Find the stock named Nintendo (ticker: NTDOY).Other investments may not be used for this assignment. (4) Gatherthe information you need from severaldifferent websites.

SECURITY ANALYSIS: Security analysts should do detective work to discover valuable information. For more information consult: (A) Google.com;(B)www.marketwatch.com; (C)www.morningstar.com; (D) www.valueline.com; (E) Vanguard’s web site; and, (F) Nintendo’s web site.

REQUIRED INFORMATION:Write double-spaced type-writtenreport that has a total of fourpagesof your writing.Where is each corporation’s world headquarters located and where is its common stock listed? For VFINX, what is its assets under management (AUM), how many different mutual funds Vanguard Group (not VFINX)manages, the goal of VFINX’s portfolio manager, annual turnover rate, the portfolio manager’s education, the fund’s management fee, load fee, exit fee, and 12b-1 fee.What was NTDOY’s largest short-term gain during 2016-2020? Explain NTDOY’shistory and forecast what you predict lies ahead for the stock. How haveVINFX and NTDOY performed over the last few decades? Shouldyou manage these investmentsactively or passively?

REPORT: Writea total of [(two pages for each investment) times (two investments) equals] fourdouble-spaced typewritten pages which follow your cover page. Please do not insert any graphs in your report, you are being graded on your writing skills.Noplagiarism will be tolerated.Staple your(1+2 + 2 =) five-pagedocument together with page numbers typed at the bottom of each page.You may list your References on a sixth page if you wish, or use footnotes instead. Print your name and Prof’s name on every page in case Prof. accidently drops all of papers and the pages become separated and mixed up.

DUE DATE: September 23-26 inclusive, 2020. Your grade will be penalized onepoint per day if you turn in your paper before September 23rd or after September 26th.

THREE GOALS OF THIS TERM PROJECT:  (A) Writing: Learn to write a report in a professional manner.  (B)  Investing: Learn the difference between active and passive investment management.   (C)  Financial data:  Become more familiar with the investments software and data available on the internet.

## Research Paper Writing-Civil engineering

Civil 313 Earthquake Homework Problems 2A – 2F

See the followingfloor plan sketches and elevation of the proposed new 3 story classroom building on campus at CSU Chico.

The gravity framing consists of the following:

Roof:             Plywood on 2×12’s spanning to wood trusses, which span to the exterior walls.

3rd Floor:       Concrete-filled metal deck spanning between wide-flange steel beams which span to wide-flange steel girders or columns in the perimeter walls.

2nd Floor:      8″ concrete slab spanning to 24″ x 24″ square concrete columns below.

The lateral force resisting system consists of concrete shear walls extending the full height of the building (these walls are not shown on the framing plans).  See the attached sketches for floor plans and framing details, an elevation, and shear wall layout.

2A.     Calculate the following (see the additional information below):

A.        The seismic weight tributary to each floor, wx.

B.        The seismic weight of the entire building, W.

C.        The coordinates of the center of mass at the roof only.

2B.     Calculate the total building seismic base shear, V, in the north-south direction only. Assume the total seismic weights of each floor are: Roof = 150 kips, 3rd Floor = 700 kips, 2nd Floor  900 kips (note these are not the answers to 2A!). Use Ss = 0.616, Sl = 0.274. The lateral force resisting system is a Building Frame System with special reinforced concrete shear walls. The building capacity is more than 500 students (Risk Category Ill) and the soil site class is D.

1. Assume the exterior walls weigh 20 pounds per square foot (psf).
2. Neglect the weight of the shear walls (you wouldn’t do this in real life!).
3. Use the unit weights on the Material Weights handout.
4. Add a uniform partition seismic mass of 10 psf for interior walls over the entire area on each floor.
5. Provide an additional 5 psf at the roof and at each floor for weights of mechanical and electrical systems, sprinklers, etc.
6. Wide flange beams and columns are designated “W14x90”, where 14 stands for the nominal depth of the “I” beam in inches and the “90” indicates its weight in pounds per lineal foot.