NIMH and University of North Carolina at Chapel Hill will review and prioritize all requests for screening.

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1 Investigator Edition This document is a tutorial for requesting compound screening by the Psychoactive Drug Screening Program (PDSP) at University of North Carolina at Chapel Hill. This service provides screening of novel psychoactive compounds for pharmacological and functional activity at cloned human or rodent CNS receptors, channels, and transporters. Screening of compounds is provided to qualified academic investigators at no cost. NIMH and University of North Carolina at Chapel Hill will review and prioritize all requests for screening. STEP 1 of 6: Register with the Online PDSP Database In order to access the assay request manager, you must be registered with our system. If you have already received an with your username and password you can go to step 2. If you don t have an account, you can register by going to: You will note that PDSP is using Hyper Text Transfer Protocol Secure (HTTPS), which is the secure version of HTTP, the protocol over which data is sent between your browser and the website that you are connected to. The 'S' at the end of HTTPS stands for 'Secure'. It means all communications between your browser and the website are encrypted. Once you have an account you are registered as an Investigator and you are ready to access our PDSP Online Database. You can login here: When logging in, if you incorrectly log in three times in a row, your account will be locked to prevent brute force password cracking techniques. You can me if you think your account has been locked or if you forget your password. This is another one of the many ways in which we are trying to protect your account and your data. Once you login, you should see drop down menus below the yellow navigation buttons. There should be 4 menus, Edit, View, Admin, and Help. Please Estela Lopez (EstelaLopez@unc.edu) if you do not see the menus when you login. These menus work in the same fashion as windows menus. Selecting a particular menu will expand the drop down, and any selection you pick will take you to that page. The yellow navigation buttons ( PDSP Home, Ki DB Home, Tracer Database, etc) are NOT part of the PDSP Online Database. They are simply there for your convenience and are other sites related to the PDSP and Dr. Bryan Roth s laboratory.

2 STEP 2 of 6: Add an assay request To start the assay request process, select the Add an Assay Request from the Admin menu. Once you click on Step 1 the following form will be displayed, You need to fill all the information out on this form, then click on Add Assay Request. Please note that the system will store your requests and you can access them at any time. You

3 do NOT have to do this whole tutorial all at once. Once you have added a request you can always edit it later and pick up where you left off. Note that the request has now been saved in the system but it has NOT been submitted. You are free to change anything and everything about the request until it is submitted to us. To continue working with the assay request, click on the add compounds link. STEP 3 of 6: Add compounds to your request In order to have your compounds screened, they must be added to an assay request. Each request can have any number of compounds. You will then request assays for each compound. Each compound you add to this request MUST have a name, and molecular weight. The chemical formula and the compound s structure are optional. This is NOT required. Adding an Image of a Compound Images must be one of:.bmp,.gif,.jpg,.jpeg,.tif Images must be small, shoot for no more than a couple hundred K. (< 500K) Please include images that include only one compound. To add an image, click on the Browse button. Select the image you want to include from the file dialog, then click Open. The path to the image now appears in the text field. When you have completed filling out the compounds details you have two choices: Submit Compound and Add Another Compound or Submit Compound. Submitting multiple compounds If you want to add another compound after this one, you can click the Submit Compound and Add Another Compound button. This will simply add your compound, and reload the Add

4 Compound page. Clicking on the Submit Compound and Add Another Compound brings up the next screen: Note that the assay request s current compounds are listed. This is a way to double-check that everything was added correctly. If you don t see your compound in the list, something went wrong and you should see an error message. You can contact Estela Lopez (EstelaLopez@unc.edu) if this happens. Adding One Compound at a Time If you click the Submit Compound button you will only be adding one compound at this time. In this example I ve added another compound, called tutorialcompound2 and clicked on the Submit Compound button. The resulting screen would look like: Note that both my compounds show up in the list. From here we can now go to the Compound Manager which is the main screen for viewing and editing assay request compounds. We select which receptors we would like to screen the given compound at by adding receptors to the compound. We can add receptors to single compounds or to multiple (even all) compounds in

5 the current request. This is all done through the Compound Manager. STEP 4 of 6: Assign receptors to your compounds The compound manager has two main parts explained below, they are: 1. Compound Selection 2. Assay Selection This page is mainly for selecting which receptors a compound or compounds will be screened at. Each selected compound will be screened at whichever receptors are selected. Compound Selection

6 This section of the page has 3 purposes: Select compounds to be used in conjunction with the assay selection section Delete compound(s) from assay request Edit a single compound There are two list boxes; the one on the left is the list of compounds you have added to this assay request. The list box to the right of the buttons is only used when ONE compound is selected, and it will list the assays that you have requested for that compound. The 4 buttons in the middle of this section operate as follows: Edit Selected Compound: You can edit a compound by selecting it and clicking on the Edit Selected Compound button. You can only edit ONE compound at a time. Editing a single compound will give you the opportunity to change the compound name, molecular weight, chemical formula, and structure image (if applicable). Detailed explanations on editing a compound are on page 3. Remove Selected Compounds: You can permanently remove a compound from the current assay request by highlighting it and clicking the Remove Selected Compounds button. The system will ask you to make sure that you mean to delete the compound(s). Click OK to delete the compound. Select All/Unselect All Compounds: You can multi-select compounds by holding the control key and clicking on individual compound names. Alternatively, you can click the Select All Compounds button to select all compounds. Once you have more than one compound selected, the right hand list box will be blank because each compound COULD have different assays requested. You can unselect all compounds by clicking the Unselect All Compounds button. After your compounds are selected using these tools, you will request assays. Receptor Selection

7 The assay selection section is used in conjunction with the compound selection section. You are trying to request assays for the compounds selected in the compound selection section. This section is useful if you want to apply the same assays to multiple compounds. You can also specify which type of experiment you are requesting. The default experiment type is primary binding assay. ONLY UNDER SPECIAL CIRCUMSTANCES would you choose secondary binding or functional assays. Note: You can only specify ONE experiment type per compound. You cannot have some primary binding assays and others to be secondary binding assays for the same compound. So when you select an experiment type here, you are making that the experiment type for all of the selected compounds. It is possible to have one compound have primary binding and another compound to have secondary binding. There are 3 ways to assign assays to the selected compounds by selecting the appropriate radio button: 1. Screen at Standard Receptors - This is the default choice and is the set of receptors we generally screen compounds at. 2. Screen at Existing Receptor Group Here you can pick a group of receptors. When you select this radio, whichever group is the current selection, for instance Adenosine, the contents of that group will be populated in the current assay list. 3. Screen at a Custom Set of Receptors With this option, you are essentially making your own group of receptors. Screening at a Custom Set of Receptors You are now using the two list boxes. When you select the Screen at a Custom Set of Receptors radio button, the list on the right will be cleared. This serves as the current list of assays. When you click on the Assign Assays to Selected Compounds button, you are requesting the selected compounds to be screened at all the receptors in the current assays list. You can change the current assay list using the 4 buttons:

8 Clicking Add Receptor(s) to List will append the receptors selected in the left list box to the current receptors list. If a receptor is already in the current assays list it won t be added twice. Clicking this button will simply make the current assays list the same as the receptor list on the left. Every receptor we have will be in the current assays list. This button will remove any receptor that is selected in the LEFT hand list of receptors from the current assays list. Clicking this button will clear the current assays list. For example, select Screen at a Custom Set of Receptors, this changes the radio selection from Screen at Standard Receptors and will clear the current assays list. If you wanted to request assays at 5ht1a, 5ht1b, and 5ht1d, select those three receptors and click Add Receptor(s) to List. They are now listed in the current assays list box. Now if I wanted to remove 5ht1a and 5ht1d from the list of current receptors, I would select them as shown below:

9 Then click the Remove Receptor(s) From List button, which results in: Maybe you want both compounds screened at 5ht1a, 5ht1b, 5ht1e, 5ht3, and 5ht6. To do that, I would first select both compounds. Then select the appropriate receptors from the receptor list and click Add Receptor(s) to List to add them to the current assays list. Finally, click the Assign Assays to Selected Compounds button.

10 Once I click Assign Assays to Selected Compounds : No matter what receptors each compound previously had assigned to them, they will be REPLACED by the list of current assays (right hand list box in the assay selection section). The resulting screen will look like: Note that the compounds affected are printed and the assays those compounds are requested to

11 be screened at are printed. TutorialCompound1 and tutorialcompound2 are set to be screened at 5ht1a, 5ht1b, 5ht1e, 5ht3, 5ht6. Clicking on Return to Manage Compounds will bring you back to the previous page. Now when you select one of the compounds, you will be able to see which assays a given compound has by clicking on its name. The compound selection section looks like: Editing a Single Compound Now, if you want to change a single compound s details, you can select its name from the compound list and click the Edit Selected Compound button. You can only edit ONE compound at a time with this button. Suppose I want to edit tutorialcompound2. I select it from the list on the left and click the Edit Selected Compound button. The page is shown below. If you added an image to the compound, it should be shown on this page. If it isn t, you can Estela Lopez (EstelaLopez@unc.edu). You can change (or add an image) to the compound by clicking the Browse button. Alternatively, you can remove the picture by typing NULL into the Change Image to: text box. Also note that the assay selection section of the compound manager is present on this page. The same rules apply here except assays are only assigned to this compound, instead of multiple compounds.

12 So if you wanted to add (or remove) a few receptors, follow the instructions for the receptor selection section of the compound manager discussed previously. Maybe we want this compound to be screened at the D1 and D5 dopamine receptors in addition to the current ones. I

13 would select D1 and D5 from the list on the right, click the Add Receptor To List button and they would be appended to the list on the right. The list on the right is the current receptor list for the current compound. I would then click the Update Compound button. Note that the list of assays now includes D1 and D5. And when we return to the compound manager, if I were to click on tutorialcompound2, the new list of requested assays would show up in the right hand list box of the compound selection section. I would repeat the above steps, adding compounds as necessary and requesting assays until I am satisfied. I would then return to the main assay request manager screen. You can reach this screen by clicking on the Return to Assay Request Manager link which is available on most screens. STEP 5 of 6: Submit your request Your compounds will not be assigned PDSP compound numbers until your request has been submitted and approved. After being assigned compound numbers and the samples being received at PDSP, then the experiments will be scheduled. You can then check the system at any time to see if your results have been posted. You can submit your request from the assay request manager main page shown below.

14 So there is now a new assay request named tutorialcompounds which I created for this tutorial. For each assay request we have a number of fields (columns): Submit Request: This column is used to submit requests. If you want to submit an assay request, click on the appropriate checkbox. There will be a confirmation dialog box just to be sure. If you click OK, your request will be submitted. The status of the request will then be Submitted and you will not be able to edit your request. Any changes will need to be made via or by phone. Name: This is just the name of the request. If you click on this link you will be able to edit the request s name, the grant number, type of project, the receptor/enzyme activity, and the rationale for testing the compounds. These were the first fields you could fill out when you added the request. Status: Status definitions are listed below the table, and all requests are New until you submit them. Once you submit a request, it must be approved by multiple parties. If no one has approved or denied your request yet, it will have a status of Submitted. If one (or more) but not all, of the parties have approved your request, its status will be Pending. If ANY one of the parties has denied your request, its status will be Denied. You will also receive an explaining why your request has been denied and what you can do about it. If all the parties required have approved your request, it will have a status of Approved. You will also receive an in this case explaining what happens next. Compounds: In this column you have two links, Add and Edit. Clicking Edit will take you to

15 the Compound Manager again. Clicking Add will give you the chance to add more compounds to the assay request. Both forms are described earlier in the tutorial. Comments: This column is used to add comments to your request and also to view comments others have added to your request. Some possibilities here are if your request is denied, there might be a comment added that explains why. Or you might want to comment that you want some receptors screened one way, or at a certain concentration, etc. Anything you want to say extra that you couldn t put anywhere else in the request. Date Modified: This is the last day that ANYONE modified your request in any way. Step 6 of 6: Find out your PDSP numbers and watch for experiment results Once your request has been approved, your compounds are all assigned PDSP numbers. You then use these numbers to check for experiment results. PDSP numbers are viewable via the compound list. You can reach the compound list from the View menu. Clicking the View Compound List button with the radio for All compounds will list all of the compounds that have been approved. The list will include the original compound name, its structure, molecular weight, the assays it is going to be screened at, and the PDSP compound number it has been assigned. Results are viewable via the various Result Lists. These are all available from the View menu.

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