Exhibit 10.5


AGREEMENT
BETWEEN

ALBERT

EINSTEIN HEALTHCARE NETWORK

AND

LINGUAGEN


This Agreement is entered
into by and between Linguagen Corp., a Delaware corporation (“Linguagen”),
and Albert Einstein Healthcare Network (“AEHN”), a non-profit
corporation in Philadelphia, PA, as of September 30, 2005 (the “Effective
Date”).


WHEREAS, Linguagen wishes
to use AEHN’s animal facilities located at 5501 Old York Road, Philadelphia, PA
19141 (“Facility”) for executing Linguagen Protocols (as defined below)
and AEHN has unoccupied space which is available in its Facility that may be
leased to Linguagen for its use.


WHEREAS, Linguagen in
conjunction with its use of the Facility for performing its Protocol proposes
to retain the services of AEHN to care and maintain its laboratory animals (“Services”).


NOW, THEREFORE, in
consideration of the mutual covenants contained herein, the parties hereby
agree to be legally bound as follows:


ARTICLE
I – STATEMENT OF WORK


AEHN shall make the
Facility available and perform Services for Linguagen in connection with
Linguagen’s protocol attached hereto as Exhibit A and any additional
protocols that Linguagen may from time to time perform at the Facility
(collectively, “Protocols”). Notwithstanding the foregoing, all
Protocols must be approved by AEHN’s Institutional Animal Care and Use
Committee “IACUC”) in accordance with the “Animal Use Protocol” attached
hereto as Exhibit B. In addition to Linguagen personnel authorized to
conduct Protocols at the Facility, Linguagen shall be allowed to have visitors
at the Facility, provided, that such visitors shall be supervised by either
Linguagen personnel or AEHN personnel at all times.


ARTICLE
II – PERIOD OF PERFORMANCE


This Agreement shall
begin on September 30, 2005 and shall not extend beyond September 30, 2006
unless the period is extended in writing by mutual agreement of the parties.


ARTICLE
III – CONSIDERATION


Linguagen shall pay AEHN
in accordance with the budget attached hereto as Exhibit C (“Budget”).
AEHN shall submit a detailed invoice for all amounts payable under this
Agreement on the last business day of each month in which the Facility is used
by Linguagen or on which AEHN performs Services. Payment of any undisputed
amounts shall be made within thirty (30) days of receipt of a proper invoice.
Checks shall be made payable to Albert Einstein Healthcare Network and sent to:


Attention: Dan Lyons

Office for Research and Technology Development

Albert Einstein Healthcare Network



 1
 









5501 Old York Road, Korman 100

Philadelphia, PA 19141


AEHN Tax ID number is
23-2290323


ARTICLE
IV – INTELLECTUAL PROPERTY RIGHTS


A.                     Linguagen
shall have worldwide right, title and interest in and to any and all
inventions, original works of authorship, findings, conclusions, data,
discoveries, developments, concepts, improvements, trade secrets, techniques,
processes and know-how, whether or not patentable or registrable under patent,
copyright or similar laws, that AEHN (through its employees) may solely or
jointly conceive, develop or reduce to practice, or cause to be conceived,
developed or reduced to practice, in the performance of the Services
(collectively, the “Inventions”), including any and all moral rights and
intellectual property rights inherent therein and appurtenant thereto,
including, but not limited to, all patent rights, copyrights, trademarks,
know-how and trade secrets and the rights to apply for the same (collectively, “Intellectual
Property Rights”.


B.                       Upon
the request and at the expense of Linguagen, AEHN shall execute and deliver any
and all instruments and documents and take such other acts as may be necessary
or desirable to document the assignment and transfer described in Paragraph A
of Article IV herein or to enable Linguagen to secure its rights in the
Inventions and Intellectual Property Rights relating thereto in any and all
jurisdictions, or to apply for, prosecute and enforce Intellectual Property
Rights in any and all jurisdictions with respect to any Inventions or to obtain
any extension, validation, re-issue, continuance or renewal of any such
Intellectual Property Right. Without limiting the foregoing, AEHN shall
disclose to Linguagen all pertinent information and data with respect thereto
and shall execute all applications, specifications, oaths and all other
instruments which Linguagen deems necessary in order to apply for and obtain
such rights and in order to assign and convey to Linguagen the sole and
exclusive right, title and interest in and to such Inventions and any
Intellectual Property Rights relating thereto. If Linguagen is unable for any
other reason to secure AEHN’s signature to apply for or to pursue any
application for any United States or foreign patent, trademark, copyright or
other registration covering Inventions assigned to Linguagen hereunder, then
AEHN hereby irrevocably designates and appoints Linguagen and its duly
authorized officers and agents as AEHN’s agent and attorney in fact, to act for
and in AEHN’s behalf and stead to execute and file any such applications and to
do all other lawfully permitted acts to further the prosecution and issuance of
letters patent or trademark, copyright or other registrations thereon with the
same legal force and effect as if executed by AEHN.


ARTICLE
V – ADMINISTRATIVE PERSONNEL


The contract
administrator for AEHN is Mary Klein and the technical administrator for AEHN
is Dan Lyons. The above personnel at AEHN can be contacted at (215) 456-7215.
The technical and contract administrator for Linguagen is Heather Kaminski, who
can he contacted at (609)-860-1500.



 2
 









ARTICLE VI – TERMINATION


Either party may
terminate this Agreement upon thirty (30) days written notification to the
other party.


A.                     In
the event of termination by Linguagen, AEHN will be reimbursed for any non-
cancelable obligations properly and reasonably incurred prior to the date of
notice of termination


B.                       In
the event of early termination without “cause” by AEHN, AEHN will provide
general cooperation and assistance in the transfer of all animals housed at
AEHN’s Facility and shall reimburse Linguagen for all reasonable costs
associated with such transfer. For purposes of clarity “cause” shall mean
Linguagen’s failure to perform any of the following obligations: (a) to make
payments of any undisputed amount due under the terms of this agreement within
thirty (30) days of receipt of the proper invoice, (b) to comply with the Guide
for the Care of Use of Laboratory Animals included in the Animal Use Protocol
attached hereto as Exhibit B, (c) to comply with AEHN’s attending veterinarian
orders and/or instructions, or (d) to properly monitor Linguagen personnel at
the Facility for any condition or behavior which might reasonably and adversely
affect the working environment at the Facility, or (e) to replace disruptive or
offending Linguagen personnel from the Facility upon the reasonable request of
AEHN.


ARTICLE
VII – INDEMNIFICATION


A.                     Linguagen
shall defend, indemnify and hold harmless AEHN, its trustees, officers, agents,
and employees from any and all liabilities, loss or damages, costs, judgments,
claims, actions or suits (“Claims”) arising out of or in connection with
the activities carried out pursuant to this Agreement. Notwithstanding the
foregoing, Linguagen shall not be obligated to indemnify for any Claim where
(a) such Claim is caused by gross negligence or willful malfeasance of AEHN,
its IACUC, trustees, officers, agents, or employees, (b) Linguagen is not
promptly notified of such Claim, (c) Linguagen does not have sole control over
the defense and settlement of such Claim, or (d) Linguagen does not have the
right to select defense counsel and to direct the defense or settlement of such
Claim.


B.                       AEHN
shall defend, indemnify and hold harmless Linguagen, its, officers, board
members, agents, and employees from any and all Claims arising from (a) a
material breach of this Agreement by AEHN, or any other trustee, officer,
agent, or employee of AEHN, or (b) gross negligence or intentional misconduct
on the part of AEHN or any trustee, officer, agent or employee of AEHN.
Notwithstanding the foregoing, AEHN shall not be obligated to indemnify for any
Claim where (a) such Claim is caused by gross negligence or willful malfeasance
of Linguagen, its officers, agents, or employees, (b) AEHN is not promptly
notified of such Claim, (c) AEHN does not have sole control over the defense
and settlement of such Claim, or (d) AEHN does not have the right to



 3
 









select defense counsel and to direct the defense or
settlement of such Claim.


C.                       The
indemnifying party shall provide a diligent defense against or settlement of
any claims brought or actions filed with respect to the subject of the
indemnity contained herein, whether such claims or actions are rightfully or
wrongfully brought or filed. The indemnifying party shall have the right to
settle Claims at its sole expense, provided, that the indemnifying party may
not settle a claim without the written consent of the indemnified party, which
consent shall not be unreasonably withheld or delayed.


D.                      The
indemnitee shall cooperate with indemnifying party and its legal
representatives in the investigation and defense of any Claim under this
Agreement. In the event a Claim is asserted, the indemnitee shall have the
right to select and to obtain representation by separate legal counsel. If the
indemnitee exercises such right, all costs and expenses incurred by indemnitee
for such separate counsel shall be borne solely by the indemnitee.


ARTICLE
VIII – USE OF NAMES


AEHN and Linguagen will
obtain prior written permission from each other before using the name, symbols
and/or marks of the other in any form of publicity in connection with the
Agreement. Notwithstanding the foregoing, such written approval shall not be
required for disclosure (a) by either party of the existence of this Agreement,
(b) Linguagen’s use of the name, symbols and/or marks of AEHN as the operative
site where the Protocol was performed, or (c) as required by law.


ARTICLE
IX – ENTIRE AGREEMENT


This Agreement
constitutes the entire agreement between the parties regarding the subject
matter herein. Any changes to this Agreement shall be made in writing and must
be signed by authorized representatives or designees of each party. No waiver
of any rights under this Agreement shall be effective unless in writing signed
by the party to be charged. A waiver of a breach or violation of any provision
of this Agreement will not constitute or be construed as a waiver of any
subsequent breach or violation of that provision or as a waiver of any breach
or violation of any other provision of this Agreement.


ARTICLE
X – MISCELLANEOUS


A.                     All
notices and other communications required or permitted hereunder shall be in
writing and deemed to have been given when hand delivered, sent by facsimile or
mailed by registered or certified mail or overnight courier with tracking
capabilities, as follows or as a party may otherwise notify to the other in
accordance with this Section 5.2 (provided, that such notice of change
of address or recipient shall be deemed given only when received at the address
listed below.


B.                       This
Agreement shall be governed and interpreted in accordance with the law of the
Commonwealth of Pennsylvania without giving effect to any conflict of laws



 4
 









provisions. The parties agree that any dispute or
controversy arising out of or relating to any interpretation, construction,
performance or breach of this Agreement may be brought in the United States
District Court in Pennsylvania, or if such court does not accept jurisdiction
or will not accept jurisdiction, in any court of general jurisdiction in the
Commonwealth of Pennsylvania.


C.                       If
any provision of this Agreement or application thereof to anyone or under any
circumstances is adjudicated to be invalid or unenforceable in any
jurisdiction, such invalidity or unenforceability shall not affect any other
provision or application of this Agreement which can be given effect without
the invalid or unenforceable provision or application and shall not invalidate
or render unenforceable such provision or application in any other
jurisdiction.


D.                      This Agreement may be executed in two counterparts and by facsimile,
each of which shall be deemed to be an original as against any party whose
signature appears thereon, but both of which together shall constitute but one
and the same instrument.


[SIGNATURE PAGE FOLLOWS]



 5















IN WITNESS WHEREOF, the parties hereto have executed this Agreement as
of the 30 day of September, 2005.


	
  LINGUAGEN CORP

  2005 Eastpark Boulevard

  Cranbury, NJ 08512-3515

  	
   

  	
  ALBERT EINSTEIN

  HEALTHCARE NETWORK

  Office for Research and Technology

  5501 Old York Road, Korman 100

  Philadelphia, PA 19141

  



 


	
  /s/ Scott Horvitz

  	
   

  	
   

  	
  /s/ Mary Klein

  	
   

  
	
  Scott Horvitz

  Chief Financial Officer

  	
   

  	
  Mary Klein, PhD

  Director, Office of Research and

  Technology Development

  



 


	
  9.30.05

  	
   

  	
   

  	
  10/6/05

  	
   

  
	
  Date

  	
   

  	
  Date

  



 




	
  Approved

  	
  Illegible

  	
   

  
	
  Finance

  	
  9/30/05

  	
   

  
	
  Legal

  	
  Illegible

  	
   

  
					






 



 1















Exhibit A

Animal Use Protocol


	
  Albert
  Einstein Healthcare Network

  
	
  Animal
  Care Committee

  
	
  ANIMAL
  USE PROTOCOL APPLICATION

  



A.                       Administrative
Information


1.                         INDICATE
YOUR SELECTION BY CHECKING ALL THAT ARE APPROPRIATE.


	
  x

  	
   

  	
  New:

  
	
  o

  	
   

  	
  Renewal:

  
	
  o

  	
   

  	
  Revision:

  
	
  o

  	
   

  	
  Pilot Study:

  
	
  o

  	
   

  	
  Other:

  



 


2.                         PROTOCOL
TITLE:                          Lickometer
Taste Testing in Mice


3.                         PRINCIPAL
INVESTIGATOR(S):                         Kyle Palmer, PhD


4.                         PHONE:                       609-860-1500
x226


5.                         BEEPER:                   609-858-8797


6.                         E-MAIL:                       kyle.palmer@linguagen.com


7.                         DEPARTMENT/LOCATION:
:  Linguagen Corp.


8.                         DEPARTMENT
CHAIRPERSON:


9.                         TECHNICIAN(S)/CO-INVESTIGATOR(S)DEPT.(S)           TITLE(S)            PHONE


Heather Kaminski, Senior Research Associate     609-860-1500 x 216


10.                  CONTACT PERSON (S) FOR AFTER NORMAL WORKING
HOURS: List name(s) to contact in the event there is a need to reach
a protocol participant for animal care problems that occur after normal working
hours or when the Principal Investigator is not available. Heather Kaminski.
908-578-6859


B.                       PROJECT
DESCRIPTION


1.                         START
DATE:                       October
2005


2.                         FINISH
DATE:                        October
2008



 1
 









3.                         PURPOSE: To
ascertain difference in taste preference of solutions containing either a
standard tastant or a novel compound tastant in mice.


4.                         ABSTRACT: In order to determine taste preference in mice we
will be using a behavioral study known as a lickometer test. This test will
allow us to determine if there is an aversion to a liquid substance based on
the taste of the animal. We want to use this assay in order to determine if a
substance can be used to alleviate an aversive bitter taste to mice. These
substances will be important because they can potentially be used to modify
bitter taste in human which can help patients comply with oral regimens


5.                         LAYMAN’S
PARAGRAPH: Humans have an aversion to bitter taste which is beneficial in
avoiding toxic compounds. However, when prescribing oral suspension of drugs,
especially in pediatric and geriatric patients, aversion the the bitterness of
the drugs leads to a lack of compliance. In order to monitor substance which
may block bitter taste we will be using mice to measure aversion. This aversion
is measured through a behavioral apparatus called a lickometer. The lickometer
is a modified mouse cage which allows a mouse to “taste” a small amount of
liquid during a short period of time. The number of licks or tastes the mouse
takes is recorded by computer. This number is then used in comparison to a
nonbitter compound, usually water, to determine if the compound does not taste
well to the animal. In this manner one can add additional compounds to bitter
ones to monitor any effect they may have in alleviating the aversive taste to
mice.


6.                         DESCRIBE
METHODS AND PROCEDURES SPECIFIC TO THE ANIMAL USE IN THE PROTOCOL: Mice
will be tested in an automated lickometer referred to as the ‘Davis Rig’ (DiLog
Instruments, Tallahassee, FL) The Davis Rig allows presentation of up to 16
different taste stimuli within a single behavioral session, with duration and
order of stimuli presentation at control of technician. The test chamber is a
rectangular cage made of plastic with a wire mesh floor. An opening in the
center of the front wall gives animals access to a sipper tube that contains a
taste solution. Fluid access is controlled by a computer operated shutter. The
amount of licks will be monitored for each session using a high frequency AC
contact circuit. Mice will be water deprived ON (~15 hours) prior to the
initial test session of the week. Mice will have access to water ad libitum in
their own cages except during the initial ON restriction. The first session
will allow mice to learn that water is available to them through the sipper
tube by exposing the animal to the open sipper tube containing distilled water
for 20 minutes. The next session will allow the mouse to learn brief-access
trials. The same distilled water will be used in this trial but instead of 20
minutes the animal will be exposed to 15 second intervals of open shutter until
30 trials were reached. Testing will occur over the next three days in three
identical sessions. The 30 trials will now be broken into 5 blocks of 6 trials
each where each of the 6 trials contains a different tastant solution. Mice
will be kept in the rigs no more then 30 minutes per trial. Number of licks
again will be monitored and compared to a set of standard compounds including
quinine, ranitidine, and cyclohexamide. Mice will be weighed periodically to
monitor weight loss and mice losing more then 20% body weight will be removed
from testing. After a 5 day test mice will be given a rest period before
another test begins.



 2
 









C.                       ANIMAL
SELECTION


1.                         SPECIES:               Mus musculus


2.                         STRAIN:                   C57/BI6


3.                         SEX:     F/M


4.                         AGE
AND/OR WEIGHT RANGE:                             18-20g


5.                         SOURCE: Taconic

PLEASE JUSTIFY THE SELECTION OF A VENDOR NOT
ON THE LIST:


6.                         JUSTIFY
THE USE OF LIVING ANIMALS: The complexity of the processes being study
cannot be duplicated or modeled in simpler systems.


7.                         JUSTIFY
THE APPROPRIATENESS OF THE SPECIES SELECTED FOR THE STUDY: This species
provides a good model for duplicating the human response to aversive bitter
tastes.


8.                         TOTAL
NUMBER OF ANIMALS TO BE USED IN THIS PROTOCOL:   120


9.                         RATIONALE
FOR DETERMINING THE NUMBER OF ANIMALS NEEDED: We will use 4 groups of ten
animals each. Group 1 will have water and compound A. Group 2 will have water
and drug B. Group 3 will have x and standard A. Group 4 will be controls (water
and no drugs). Data will be compared using a Student’s “T” test. From previous
experiments, we estimate that ten will be an adequate group size to show
significant results. Each of the 4 groups can be used for 3 (1 week) studies.


D.                       PRE-ANESTHESIA
/ ANESTHESIA / SEDATIVES / ANALGESICS


1.                          If Sedation, analgesics, tranquilizers. and /or
anesthesia is (are) used, please Identify below. (see appendices for
recommendations)


a.


	
  PRE-ANESTHETIC OR

  SEDATIVE AGENT

  	
   

  	
  DOSE

  	
   

  	
  ROUTE

  	
   

  	
  VOLUME

  	
   

  	
  FREQUENCY

  OF DOSING

  	
   

  	
  DURATION OF

  TREATMENT

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  N/A

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
     

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  



 


b.


	
  ANESTHETIC DRUG

  	
   

  	
  INITIAL

  DOSE

  	
   

  	
  SUPPLEMENT

  DOSE

  	
   

  	
  ROUTE

  	
   

  	
  EXPECTED TIME UNDER

  ANESTHESIA

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  N/A

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
     

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  




 3
 









c.


	
  1.               POST-ANALGESIC

  OR SEDATIVE

  AGENT

  	
   

  	
  DOSE

  	
   

  	
  ROUTE

  	
   

  	
  VOLUME

  	
   

  	
  FREQUENCY

  OF DOSING

  	
   

  	
  DURATION OF

  TREATMENT

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  N/A

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
     

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  



 


2.                         ANESTHESIA
MONITORING (check all techniques that will be used for assessing adequate
anesthesia level. Also indicate the frequency of monitoring.


	
  o

  	
   

  	
  Response to toe/skin pinch - frequency:

  
	
  o

  	
   

  	
  Palpebral/corneal reflex - frequency:

  
	
  o

  	
   

  	
  Assessment of jaw and/or skeletal muscle tone -
  frequency:

  
	
  o

  	
   

  	
  Monitoring of physiological response:

  
	
   

  	
   

  	
  Instrumentation
  for heart rate monitoring: (Name device)

  
	
   

  	
   

  	
  Instrumentation
  for blood pressure monitoring: (Name device.)

  
	
   

  	
   

  	
  o    Other: Explain:

  



 


E.                         EUTHANASIA


1.                        METHOD OF
EUTHANASIA (See recommended methods of euthanasia in the appendices.)


o                     Overdose of anesthetic:


	
  Identify Anesthetic

  	
   

  	
  Approximate Amount

  	
   

  	
  Route

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
     

  	
   

  	
   

  	
   

  	
   

  	
   

  



 


o                     Cervical dislocation with
anesthesia or sedation:


	
  Identify Anesthetic

  	
   

  	
  Approximate Amount

  	
   

  	
  Route

  	
   

  
	
  

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
     

  	
   

  	
   

  	
   

  	
   

  	
   

  



 


o                     Cervical dislocation without
anesthesia or sedation:


o                     Exsanguination under anesthesia or
sedation:


	
  Identify Anesthetic

  	
   

  	
  Approximate Amount

  	
   

  	
  Route

  	
   

  
	
  

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
     

  	
   

  	
   

  	
   

  	
   

  	
   

  



 


x                   CO2 chamber


o                     Other:


o                     If the animals are not euthanized
at the END of the study, what will happen to them?



 4
 









2.                        STUDY END
POINT FOR ANIMALS (SELECT ALL THAT APPLY):


Experimental endpoint criteria must be specified.


	
  o

  	
   

  	
  Euthanized per protocol without treatment or
  experimentation.

  
	
  o

  	
   

  	
  Euthanized per protocol after defined period of
  time (days/months)

  
	
   

  	
   

  	
  Define
  period:

  



x                   Euthanized per protocol after
completion of the experimental protocol.


	
  o

  	
   

  	
  Euthanized per protocol at development of
  specific signs.

  
	
   

  	
   

  	
  Explain:

  
	
  o

  	
   

  	
  Euthanized per protocol when moribund.

  
	
  o

  	
   

  	
  The animal may die as a result of the
  experimental treatment.

  
	
   

  	
   

  	
  Explain
  why this is necessary:

  



 


F.                         RESTRAINT
/ DEPRIVATION


1.                         RESTRAINTS:
Describe any proposed restraint (other than short-term hand-held restraint)
to be used on awake animals.


x NO - Prolonged
restraint will not be used.


o YES - Prolonged restraint will be used.


a.                          Justification:


b.                          Description:


c.                          Duration:
State the duration of the restraint including the length, frequency and
rest periods between restraint periods


d.                          Conditioning
and monitoring: Describe steps to be taken to condition the animals to the
restraining device and assure the comfort and well being of the animals.


2.                         DEPRIVATION
(Food or water restriction)


o NO - Food or water deprivation will not
take place.


x YES - Food or water deprivation will take
place.


e.                          Justification:                                 in
order for the animals to adopt proper behavior in the lickometers a motivation
is needed. The motivation in this case is thirst. The motivation is not
possible without the deprivation.


f.                            Description:   Mice will be water deprived
ON (~15 hours) prior to the initial test session of the week. The first session
will allow mice to learn that water is available to them through the sipper
tube by exposing the animal to the open sipper tube containing distilled water
for 20 minutes.


g.                         Duration:
24 hour before study


h.                         Conditioning
and monitoring: The animals will be monitored for signs of dehydration,



 5
 









such as weight loss. Mice losing more then 20% of body
weight will be removed from the study.


G.                       PAIN/DISTRESS


1.                         CATEGORY OF RESEARCH: The investigator
should check the appropriate category(ies) of experimentation and the number of
animals used in each.


x                   A: Number of animals:    120     


The research involves either no pain
or potentially involves momentary, slight pain, discomfort or distress,
includes simple invasive procedures (e.g., injection, blood sampling),
collection of tissues preceded by standard euthanasia, behavioral testing
without stress.


o                     B: Number of animals:         


The research potentially involves
minor short-term pain, discomfort or distress which will be treated with
appropriate anesthetics/analgesics. Includes terminal anesthetic surgery, minor
survival surgery with anesthesia and without significant postoperative pain
(e.g., biopsy) and/or implantation of minor chronic catheters (e.g., femoral
arterial and venous catheters, flow probes, etc.) and/or short-term physical
restraint (<60 minutes) of awake animals and/or induction of minor
behavioral stress.


o                     C: Number of animals:         


The research involves procedures
potentially inducing moderate pain, discomfort or distress which will be treated
with appropriate anesthetics/analgesics and/or chronic maintenance of animals
with a disease/functional deficit, includes major survival surgery with
anesthesia (e.g., orthopedic surgery on major skeletal components, bowel
resection, cardiac surgery, adrenalectomy) and/or inducement of a functional
deficit (e.g., non-painful tumor inducement, inflammation induced by adjuvants,
inducement of glaucoma, heart failure inducement), and/or physical restraint
(>60 minutes) of awake animals and/or induction of more than minor
behavioral stress.


o                     D: Number of animals:         


The research potentially involves
pain, discomfort or distress (greater than that attending routine injection)
which cannot/will not be alleviated through the administration of appropriate
anesthetic, analgesic or tranquilizer drugs. Examples include pain research,
radiation testing, toxicity testing and carcinogenesis experiments where
animals become moribund.


In order for a protocol to qualify as
category A, B or C, appropriate anesthetics or analgesics must be used if the
animal will experience more than momentary slight pain. Momentary slight pain
is defined as pain no greater than the level and duration of pain attending a
routine injection. Alternately, the animal must be immediately euthanized upon
evidence of such pain or the protocol is classified as category D.


H.                       REGULATORY
QUESTIONS


1.                         THE
ANIMAL WELFARE ACT’S “DUPLICATE EXPERIMENTS” QUESTION


a.                          Describe
the methods that were used to assure the Animal Care Committee that the animal
activities in this protocol do not unnecessarily duplicate previous
experiments.


Medline search describes similar methos but the use of
novel compounds and the direct application of these methods for taste research
is novel. Search was conducted on 08/29/05



 6
 









b.                         Indicate
resources / scientific literature used:


	
  x

  	
   

  	
  MEDLINE

  	
   

  	
  (inclusive dates  Nov 1980 -August
  2005  )

  
	
  o

  	
   

  	
  INDEX MEDICUS

  	
   

  	
  (inclusive dates                    )

  
	
  o

  	
   

  	
  AGRICOLA

  	
   

  	
  (inclusive dates                    )

  
	
  o

  	
   

  	
  EMBASE

  	
   

  	
  (inclusive dates                    )

  
	
  o

  	
   

  	
  CURRENT RESEARCH INFORMATION
  SERVICE  (inclusive
  dates                    )

  
	
  o

  	
   

  	
  Animal Welfare Information Center
  (see appendices)

  
	
  o

  	
   

  	
  Peer review by:

  
	
  o

  	
   

  	
  Consultation with an expert in the
  area of investigation (briefly describe; name, position, etc.

  
	
  o

  	
   

  	
  Other:

  



c.                         Key words
used in the search for alternatives:    Lickometer, mice, taste
testing


2.                         THE
ANIMAL WELFARE ACT’S “ALTERNATIVE” QUESTION

ALTERNATIVES TO POTENTIALLY PAINFUL PROCEDURES:


a.                         Describe
the methods that were used to assure the ACC that the Principal Investigator
considered alternatives to procedures that may cause more than momentary or
slight pain or distress to the animals. Provide a written narrative description
of the methods and resources.                   No
painful methods are being used


 


 


b.                         Indicate
resources / scientific literature used:


	
  o

  	
   

  	
  MEDLINE

  	
   

  	
  (inclusive dates                    )

  
	
  o

  	
   

  	
  INDEX MEDICUS

  	
   

  	
  (inclusive dates                    )

  
	
  o

  	
   

  	
  AGRICOLA

  	
   

  	
  (inclusive dates                    )

  
	
  o

  	
   

  	
  EMBASE

  	
   

  	
  (inclusive dates                    )

  
	
  o

  	
   

  	
  CURRENT RESEARCH INFORMATION
  SERVICE (inclusive dates                    )

  
	
  o

  	
   

  	
  Animal Welfare Information Center (see appendices)

  
	
  o

  	
   

  	
  Peer review by:

  
	
  o

  	
   

  	
  Consultation with an expert in the
  area of investigation (briefly describe; name, position, etc.

  
	
  o

  	
   

  	
  Other:

  



 


c.                         Keywords
used in the search for alternatives.:



 7
 









I.                            OTHER


1.                         Do any of
the proposed procedures involve the use of Biohazardous materials?


x                        NO - BIOHAZARDOUS materials will not be used in
or disposed of by CAF.


o                          YES - BIOHAZARDOUS materials will be used. (“Application
for the Use of Biohazardous

    Materials and Animals” must be submitted with the
Animal Use Protocol):


o                          Laser Use (Laser Task Force)


o                       Radioisotopes (Radiation Safety Committee)


o                       Carcinogens, Teratogens, Mutagens, Chemicals


o                       Transmissible/Infectious Agents/Organisms


o                       Recombinant DNA


o                       Other:


2.                         Will
tumor cells, tissue, sera or other biologies from either animal or human
sources be used in or on animal as part of the protocol?


x                     NO


o                       YES


a.                         Please
describe precautions that should be observed with materials or with live/dead
animals.


b.                        Describe
the tests used to check the tissues and cells for inadvertent contamination by
viruses or mycoplasma.


3.                         ANTIBODY
PRODUCTION


o                     Polyclonal antibodies:


o                     Monoclonal antibodies:


o                      Adjuvants used.


Adjuvant                       
(If Freund’s Complete Adjuvant is used, See ACC guidelines)

Route of administration:

Amount injected:

Site preparation:


4.                         TRANSGENIC
MICE


Will
transgenic mice be used?


o                     NO


o                     YES                                              Source:
                     


Health status: (Describe the tests used to check the
animals for inadvertent contamination by viruses or mycoplasma.)


5.                         Will
animals be transported to or through patient care areas? (This includes
hallways in Korman Research Pavilion.)


x                    NO


o                      YES                                              Identify:


Need:


Measures used to protect patients and hospital staff
from possible allergic or emotional reactions:


Describe route and methods used to move/hide animals
from view:



 8
 









6.                         Will
there be a need to house animals outside of the animal facility for more than
12 hours?


x                    NO


o                      YES           Room Number                
Building                


Please provide justification:


Describe precautions used to prevent contamination of
the area by the animals (e.g., smells, dander) and procedures to assure that
the animals are maintained in a humane manner.


J.                         PERSONNEL
QUALIFICATIONS / INFORMATION


Attach forms:


	
  Principal Investigator Qualifications in Animal
  Care and Use

  	
   

  	
  Attached: x

  
	
  Support
  Personnel Qualifications in Animal Care and Use

  	
   

  	
  Attached: x

  



 


Do you foresee any potential safety concerns beyond
the normal possibility of animal bites, animal scratches, zoonoses, and
allergens when accomplishing this protocol?


x                    NO


o                      YES


What are those concerns and what precautions will be
taken?


K.                       ASSURANCE
STATEMENTS AND SIGNATURES


“I accept and will conform to all federal and state laws and guidelines
and all institutional policies and procedures concerning the care and use of
animals in research, teaching, and testing. I understand that I have a
responsibility to notify the Animal Care Committee in writing of any
substantive changes in the proposed project, changes in personnel assigned to
work under this application, or in the number of animals or the species to be
used, prior to proceeding with any animal use in such modified projects.


	
  /s/ Illegible

  	
   

  
	
  Signature of Principal Investigator:

  



 


The information provided in the Animal Use Protocol form accurately
reflects the nature of the activities proposed.


	
  /s/ Illegible

  	
  9/26/05

  
	
  Signature of Department Chair

  



 



 9
 









Only one copy of this
page should submitted to the ACC committee coordinator. This funding source
information should not be included in the copies for distribution to the ACC.


L.                        FUNDING
SOURCES:


1.                         EXTERNAL
FUNDING


a.                         Please
provide name of the source Linguagen Corporation


It is important that researchers who are applying for
PHS funds read Pages 17-18, “Vertebrate Animals”, of the grant application
package and complete Section 2F. You may call the ORTD office at 456-7217 if
you have questions or need additional information.


b.                         Grant
Title:


c.                         Amount of
grant or contract:


d.                         Does the
award Include administrative


o                     NO


o                     YES, please
give dollar amount:


e.                         Does the
agency to which you are applying for fund require an institution approved
animal use protocol?


o                     NO


o                     YES


2.                         INTERNAL
FUNDING


x                   NO


o                     YES:


o           Department:


o           AESG:


o           Other


Most, but not all, granting agencies allow to apply for a grant before
the animal use protocol has been approved. If your protocol has been submitted,
but is not yet approved, indicate to the granting agency that the protocol is “pending”.
NIH and NSF normally allow sixty days for approval after the deadline date for
the grant. If no approval from the ACC follows within sixty days of the
deadline date for the grant application, the agency will normally discard your
grant application.


Some funding agencies (such as American Heart Association) require that
the protocol be approved before the grant is submitted. Investigators are
responsible for knowing the requirements of their particular agency.



 10














EXHIBIT B 

Animal Use Protocol Application Instructions


	
  Albert
  Einstein Healthcare Network

  Animal Care Committee

  ANIMAL USE PROTOCOL APPLICATION INSTRUCTIONS (to be used with the

  application form)

  



 


1.                         Regulating
and Accrediting Agencies


2.                         AEHN
Animal Care Committee Responsibility


3.                         Annual
Review Process


4.                         Guidelines/Instructions
for completing the Animal Use Protocol Application


A.                      ADMINISTRATIVE
INFORMATION


B.                      PROJECT
DESCRIPTION


C.                      ANIMAL
SELECTION


D.                      PRE-ANESTHESIA
/ ANESTHESIA / SEDATIVES / ANALGESICS


E.                        EUTHANASIA


F.                        RESTRAINT
/ DEPRIVATION


G.                      PAIN /
DISTRESS


H.                      REGULATORY
QUESTIONS


I.                           OTHER


J.                        SIGNATURE
STATEMENTS


K.                      PERSONNEL
QUALIFICATIONS / INFORMATION


L.                       FUNDING
(only need to furnish one copy for the Office for Research and Technology

Development)


M.                    APPENDICES


1.                         Regulating
and Accrediting Agencies


A.                       Animal
Welfare Act - Federal regulations under the Animal Welfare Act mandate
registration with the United States Department of Agriculture, Animal and Plant
Health Inspection Service, Regulatory Enforcement and Animal Care, Animal Care
Division. AEHN is registered as a Class “R”, Registered Research Facility and
as such is subject to the Animal Welfare Act regulations including unscheduled
inspections by the United States Department of Agriculture.


B.                       Health
Research Extension Act of 1985 - Under the requirements of the Health
Research Extension Act of 1985, AEHN as a recipient of Public Health Service
funding is registered with the Public Health Service, Department of Health and
Human Services, Office for Protection from Research Risks (OPRR), National
Institutes of Health and is subject to its rules and regulations. As required
by the Public Health Services (PHS), Policy on Humane Care and Use of
Laboratory Animals, AEHN must provide an acceptable written Animal Welfare
Assurance to OPRR. Our Assurance states we will follow the recommendations in
the Guide for the Care and Use of Laboratory
Animals.


C.                       Association
for Assessment and Accreditation of Laboratory Animal Care (AAALAC) - AEHN’s
accreditation provides an independent, rigorous assessment of our Animal Care
and Use Program. Maintenance of AAALAC accreditation is affirmation of the
animal program’s accountability and its efforts to promote ethical practices in
the care in the use of animals. Many granting organizations strongly recommend
an accredited animal program. Government agencies regard AAALAC accreditation
as evidence of program excellence and compliance with all policies and
regulations.











2.                         Albert
Einstein Medical Center Animal Care Committee Responsibility


All research and training involving the use of animals must have an
approved Animal Use Protocol on file with the Office for Research and
Technology Development. Animal Use Protocols are reviewed by an appointed
Animal Care Committee (ACC). The ACC is appointed by the Vice President and
General Director of AEHN as proscribed by the Animal Welfare Act and the PHS
policy. The ACC includes: scientists experienced in animal research,
non-scientists, one member not affiliated with AEMC, and a veterinarian
experienced in laboratory animals.


The ACC is also proscribed by the Animal Welfare Act and the PHS policy
to conduct semi-annual reviews of the Animal Care Program, inspections of the
animal facilities, and animal use areas. The Animal Use Protocols and the
Animal Care Committee minutes are subject to review by United States Department
of Agriculture inspectors, AEHN’s accrediting agency (Association for
Assessment and Accreditation of Laboratory Animal Care) site visitors,
officials from granting agencies (e.g., NIH) and authorized Office for
Protection from Research Risks other Public Health Service (PHS)
representatives. The ACC also investigates reported concerns regarding the
humane care and use of animals at the AEHN.


3.                         Annual
Review Process


Animal Use Protocols that have been approved by the
ACC are reviewed annually. The review process will proceed as follows:


1.                          An
annual review notice and form is sent out to the Principal Investigator
approximately two weeks prior to the protocol anniversary date. The ACC will
request the following: a notice of any substantial changes to the original
protocol, a progress summary, notice of completion, or a reason for not
completing the protocol. The annual review form should be returned within two
weeks. If the annual review form is received within that time and approved by
the Committee, an approval letter is sent to the Investigator. If more
information is required, the Investigator will be notified.


2.                          A
second notice is sent out if the annual review form has not been returned
within two weeks. The due date for this notice is also two weeks.


3.                          Final
notice is sent out with a two week due date. This allows the Investigator a
total of six weeks in which to respond to the request for an annual review.


4.                          If
no response to the final notice is received within two weeks, the protocol is
inactivated and the Investigator is notified. Animals can no longer be ordered
or housed under this protocol. Animals being housed in the Facility under the
inactivated protocol must be disposed of within two weeks.


5.                          A
new protocol form must be submitted if the Investigator decides to resume work
on an inactivated protocol.


6.                          If
you have any questions or concerns, address them to the ACC Coordinator at
215-456-7217.


7.                          After
three years, you will be requested to update your protocol by submitting a new
form. The notification process is the same as the one outlined above.


PI’s who use animals in their laboratories should expect visits by a
subcommittee of the ACC at approximately six month intervals. In addition to
looking at the facilities during the laboratory visits, the ACC members will
observe animal procedures, if in progress, discuss animal use











procedures described in the
applicable protocol(s) with the research staff, inspect drugs and materials
intended for in vivo use, and inspect research animal records


4.                         Guidelines/Instructions
for completing the Animal Use Protocol Application.


1.                          Animal
Use Protocols must be submitted to the Office for Research and Technology
Development (ORTD) for distribution to the members of the ACC. The ACC is
scheduled to meet on the second Monday of the month. Completed Animal Use
Protocol forms must be received by ORTD ten working days prior to the meeting.
The ORTD is located in the Korman Research Pavilion, Room 100 (ext. 6-7217).
The ACC Coordinator serves as the point of contact for information regarding
the ACC.


2.                          The Animal Use Protocol form must be typed.
Submit eight copies of the completed Animal Use Protocol. Please make sure
all abbreviations are clarified. The application is available on diskette.


3.                          It
will be necessary for the Principal Investigator or an appropriate substitute
to attend the ACC meeting to present a brief summary and to clarify points in
the Animal Use Protocol.


4.                          Separate
Animal Use Protocol forms must be completed for each animal use procedure and
for each species.


5.                          The
Animal Welfare Act requires the ACC to verify the qualifications of those who
are involved in the animal use of the research. To fulfill this responsibility
the Animal Care Committee asks that a record of animal use experience,
training, and intended training of all personnel be included with each Animal
Use Protocol. Please use the following forms and attach them to the protocol:


Principal
Investigator Qualifications in Animal Care and Use form 


Support Personnel Qualifications in
Animal Care and Use form


6.                          The
institutional attending veterinarian and the AEMC’s Central Animal Facility
staff (ext. .6-7159) will assist investigators in completing the Animal Use
Protocol forms, train personnel, and conduct procedures described in the
protocol.


7.                          Investigators
needing to make modifications (e.g., small increases in number of animals,
change of anesthesia, adding personnel) to their approved Animal Use Protocol
can submit a brief addendum to the ACC. Please use the Request to Modify an Approved Animal Use Protocol form.


8.                          The
Animal Care Committee will send written notice of approval, approval with
modifications, disapproval or disapproval with suggestions to allow approval.
An ACC approved Animal Use Protocol will be assigned a number. Animals can not
be used without an approved Animal Use Protocol.


A.                       Administrative
Information


1.                                                            INDICATE YOUR
SELECTION BY CHECKING ALL THAT ARE APPROPRIATE.


o                      New: The
research or training protocol has not been reviewed and/or approved previously.


o                      Renewal: This
research or training protocol was approved previously by ACC, but is
approaching the end of its 3 year approval period. Provide the previously
approved Animal Protocol Use number.


o                      Revision: A
resubmitted modified tabled protocol.


o                      Pilot Study:











o                       Other:


2.                         PROTOCOL
TITLE: Self-explanatory.


3.                         PRINCIPAL
INVESTIGATOR(S): The person with overall responsibility for the animal care
and use activities, the personnel involved with the animal procedures and
charges incurred by the animal facility.


4.                         PHONE: Self-explanatory.


5.                         BEEPER: Self-explanatory.


6.                         E-MAIL: Self-explanatory.


7.                         DEPARTMENT/LOCATION:
Self-explanatory.


8.                         DEPARTMENT
CHAIRPERSON: Self-explanatory


9.                         TECHNICIAN(S)/CO-INVESTIGATOR(S)
DEPT.(S)                                                                  TITLE(S)
                                                   PHONE


List all personnel involved in the live  animal-related activities of the
protocol. Also complete a “Qualifications in

   Animal Care and Use” form for each individual
listed.


10.                  CONTACT PERSON
(S) FOR AFTER NORMAL WORKING HOURS: List name(s) to contact in the event
there is a need to reach a protocol participant for animal care problems that
occur after normal working hours or when the Principal Investigator is not
available.


B.                       PROJECT
DESCRIPTION


1.                         START
DATE: Self-explanatory


2.                         FINISH
DATE: Self-explanatory - An approved Animal Use Protocol has a three year
life.


3.                         PURPOSE: What
are the specific scientific objectives (aims) of the research?


4.                         ABSTRACT:
The abstract should state the purpose of the study, objectives, and
describe the basic procedures or techniques. The abstract should be written in
terms understandable by a non-scientist.


5.                         LAYMAN’S
PARAGRAPH: Give a brief overview of the protocol including potential value
to human and animal health, the advancement of science, or the good of society.
The overview must be in terms that nonscientists and lay members of the
committee can understand.


6.                         DESCRIBE
METHODS AND PROCEDURES SPECIFIC TO THE ANIMAL USE IN THE PROTOCOL: (The
methods section should be written in terms understandable by a non-scientist.)


Specifically address the following
that apply to your project:


o                       Injections: (Describe the routes, amounts,
site preparation)


o                       Special diets, food, or water treatments: (Describe  the diet or water treatment, any
anticipated nutritional deficit, and special housing needs.)


o                       Blood
sampling: (Describe the techniques, volumes, frequency, and anesthesia.)
(see appendices)


o                       Invasive sampling of urine/feces: (Describe
the techniques, volumes, frequency, anesthesia.)


o                       Collection of tissues: (Describe the method
of collection and anesthesia.)


o                       Non-survival surgery: (Describe the
procedure, duration, and anesthesia)


o                       Single survival surgery: (Describe the
procedure, duration, anesthesia, and postoperative care provisions) - Aseptic
techniques must be used on most animals, including rabbits, that undergo major
survival surgery. This technique includes wearing of sterile surgical gloves,
gowns, caps, face masks; use of sterile instruments; and aseptic preparation of
surgical field. Details of the procedure should include how











instruments and materials are sterilized. Survival
surgery on rodents does not require a special facility but should be performed
using sterile instruments, surgical gloves, and aseptic procedure to prevent
clinical infections. Include: the frequency of observations during anesthetic
recovery, frequency of examinations, support therapy, criteria used to assess
pain (see appendices), and administration of pain relieving drugs.


o                      Multiple
survival surgeries involving an individual animal: (Describe the
procedures, duration, anesthesia, postoperative care, provisions for each
procedure; explain how surgeries are related.)


o                      Indwelling
catheters or implants: (Describe the site(s), type(s), and
maintenance/monitoring procedures.)


o                      Administration of drugs/reagents/cells/etc.
(Describe the agent, route of administration and frequency, anesthesia,
anticipated side effects, and monitoring procedures.)


o                      Antibody production: (Indicate the route of
administration, adjuvant use, frequency, and collection procedures.)


o                      Tumor transplantation: (Describe any
anticipated functional deficit to the animal, monitoring protocol and,
endpoints used.)


o                      Toxicity
testing: (Describe the procedure, anticipated side effects, and endpoint)


o                      Behavioral testing without significant restraint or
noxious stimuli: (Describe.)


o                      Behavioral
testing with significant restraint or noxious stimuli: (Describe testing
and provide rationale for degree of restraint or stimulus.)


o                      Procedures involving potential pain where
pain-relieving methods will not be used: (Completely
describe procedures.)


o                      Human safety
concerns or hazards: (Describe the intended use of materials which may have
possible hazards and means available to ensure the safety.)


o                      Paralytic Agents: Neuromuscular blocking
agents create a significant animal welfare concern since paralyzed animals are
difficult to monitor for anesthetic effectiveness. Scientific justification for
the use of paralytic agents must be documented. (Describe steps taken to assure
that the animal is under anesthesia or analgesic. List the agent used, dose,
route, and frequency.)


o                      Anticipated animal health changes: (Describe
possible health changes, methods to monitor changes, and contingencies for
dealing with the changes or complications. State the criteria for removing
animals from the study. See appendices: 4,5,6)


o                      Special husbandry: instructions or
considerations


C.                       ANIMAL
SELECTION


1.              SPECIES: (mouse,
rat, rabbit, dog, pig, etc.)


2.              STRAIN: (for
rodents and rabbits)


3.              SEX: (male,
female, or either)


4.              AGE AND/OR WEIGHT
RANGE:


5.              SOURCE: Name of
the vendor(s) supplying the animals.


Approved Vendor list on
file in the Central Animal Facility (x7159)


PLEASE JUSTIFY THE SELECTION OF A
VENDOR NOT ON THE LIST:.


6.              JUSTIFY THE USE OF
LIVING ANIMALS: Consider the following:


·                  The complexity
of the processes being study cannot be duplicated or modeled in simpler
systems.











·                            There
is not enough information known about the processes being studied to design
non-living models.


·                            Preclinical
studies or training in living animals are necessary prior to human testing or
use.


1.                         JUSTIFY
THE APPROPRIATENESS OF THE SPECIES SELECTED FOR THE STUDY:


What anatomical,
physiological, or other characteristics/factors did you use to select the
species in consideration of the scientific objectives of the research? Consider
the following:


·                            There
exists an immense amount of data for this species allowing comparison ....


·                            The
anatomy or physiology of this species is uniquely suited to the study because
.....


·                            This
species provides a good model for duplicating the human ....


·                            This
species is the phylogentically lowest that provides adequate size, tissue, or
anatomy for the proposed study.


1.                         TOTAL
NUMBER OF ANIMALS TO BE USED IN THIS PROTOCOL:


Estimate the total number of animals needed over the three year life of
the protocol. An account of the number of the animals ordered will be
maintained by the Central Animal Facility. To exceed the approved number, a “Request to Modify an Animal Use Protocol” must
be submitted to the ACC.


9.                         RATIONALE
FOR DETERMINING THE NUMBER OF ANIMALS NEEDED:


Investigators need to provide a clear and logical
thought process that explains why they have selected the specific number of
animals that they show on their protocol. Additionally, the investigator must
show that the number is a reasonable minimum for
the scientific goals of the study. Please do not copy the statements below. The
goal is not for investigators to have a “blanket” statement, acceptable for all
situations. The goal is for investigators to clearly communicate to the ACC
their own rationale for choosing particular numbers.


Example
statements:


“We will use 4 groups of twenty animals each. Group 1
will have x and drug A. Group 2 will have x and drug B. Group 3 will have x and
both drugs. Group 4 will be controls (no x and no drugs). Data will be compared
using a Student’s “T” test. From previous experiments, we estimate that twenty
will be an adequate group size to show significant results.”


“This is a preliminary study to develop and show the
feasibility of a new surgical technique. We estimate that it may require as
many as twenty animals for us to refine our skills technique adequately to give
the  technique a fair evaluation.”


“This study involves a very small number of animals
(6-10). The variance that will be associated with this data is unknown. The
small number of animals used in this study will provide us with an estimate of
the range and variance of this data.”


“Based on the PI’s experience using these methods in
similar studies, and reviews of related use in peer reviewed published studies,
the number of animals requested is the minimum necessary to meet the scientific
objectives of the proposed research and to provide scientifically meaningful
data.”











D.                       PRE-ANESTHESIA
/ ANESTHESIA / SEDATIVES / ANALGESICS


1.                         If
Sedation, analgesics, tranquilizers, and /or anesthesia is (are) used, please
identify.


(See Appendices for suggested methods). Also, the attending
vet is available for consultation, x7217.


2.                         ANESTHESIA
MONITORING (check all techniques that will be used for assessing adequate
anesthesia level. Also indicate the frequency of monitoring.


o                      Response to toe/skin pinch - frequency:


o                      Palpebral/corneal reflex - frequency:


o                      Assessment of jaw and/or skeletal muscle tone -
frequency:


o                      Monitoring of physiological response:


Instrumentation for heart rate monitoring: (Name device)


Instrumentation for blood pressure monitoring: (Name device.)


o                      Other: Explain:


E.                         EUTHANASIA


Euthanasia means the humane destruction of an animal accomplished by a
method that provides rapid unconsciousness and subsequent death without
evidence of pain or distress, or a method that utilizes anesthesia produced by
an agent that causes painless loss of consciousness and subsequent death.


The Animal Care Committee follows the guidelines established by the
American Veterinary Medical Association Panel (AVMA). Suggested methods are
included in the appendicies. The complete text from the AVMA Panel is available
from the CAP (extension x7159)


1.                        METHOD OF
EUTHANASIA (See recommended methods of euthanasia in the appendices.)


o                       Overdose
of anesthetic:


	
  Identify Anesthetic

  	
   

  	
  Approximate

  Amount

  	
   

  	
  Route

  	
   

  
	
     

  	
   

  	
   

  	
   

  	
   

  	
   

  



 


o                       Cervical
dislocation with anesthesia or sedation:


	
  Identify Anesthetic

  	
   

  	
  Approximate

  Amount

  	
   

  	
  Route

  	
   

  
	
     

  	
   

  	
   

  	
   

  	
   

  	
   

  



 


o                       Cervical
dislocation without anesthesia or sedation:


Justify the reason for not using
anesthesia of sedation: The AVMA states “until additional
information is available to better define the nature of the persistent EEG
activity, this technique should be used in research settings only when
scientifically justified by the user and approved by” the ACC.


o                       Exsanguination
under anesthesia or sedation:


	
  Identify Anesthetic

  	
   

  	
  Approximate

  Amount

  	
   

  	
  Route

  	
   

  
	
     

  	
   

  	
   

  	
   

  	
   

  	
   

  



 











o                       CO2 chamber


o                       Other:


o                       If the animals are not euthanized at the of the study, what will happen
to them?


2.                        STUDY END
POINT FOR ANIMALS (SELECT ALL THAT APPLY):


Experimental endpoint criteria must be specified.


o                       Euthanized per protocol without treatment or
experimentation.


o                       Euthanized per protocol after defined period of time
(days/months)


Define
period:


o                       Euthanized per protocol after completion of the
experimental protocol.


o                       Euthanized per protocol at development of specific
signs.


Explain:
(see appendices for examples of endpoint criteria)


o                       Euthanized per protocol when moribund.


o                       The animal may die as a result of the experimental
treatment.


Explain why this is necessary:


F.                                                           RESTRAINT
/ DEPRIVATION


1.                                                           RESTRAINTS:. Describe
any proposed restraint (other than short-term, hand-held restraint) to be used
on awake animals. Note: Physical restraint procedures should be used on awake
animals only after alternative procedures have been considered and found to be
inadequate. If restraint is necessary, the device must provide the animal with
the greatest opportunity to assume its normal postural adjustments, and the
duration of restraint must be minimized.


o                       NO -
Prolong restraint will not be used.


o                       YES -
Prolong restraint will be used.


a.                         Justification:
Justify the need for restraint.


b.                         Description:
Identify or describe the restraint device.


c.                         Duration:
State the duration of the restraint including the length, frequency, and rest
periods between restraint periods


d.                         Conditioning
and monitoring: Describe steps to be taken to condition the animals to the
restraining device and assure the comfort and well being of the animals.


2.                         DEPRIVATION
(Food or water restriction)


o                       NO - Food or water deprivation will not take
place.


o                       YES - Food or water deprivation will take place.


a.                         Justification:


b.                         Description:


c.                         Duration:


d.                         Conditioning
and monitoring:


G.                                                          PAIN / DISTRESS


1.                          CATEGORY OF RESEARCH: The investigator
should check the appropriate category(ies) of experimentation and the number of
animals used in each.


o                      A: Number of animals:











The research involves either no pain
or potentially involves momentary, slight pain, discomfort or distress.
Includes simple invasive procedures (e.g., injection, blood sampling),
collection of tissues preceded by standard euthanasia, behavioral testing
without stress.


o                      B: Number of animals:


The research potentially involves
minor short-term pain, discomfort, or distress which will be treated with
appropriate anesthetics/analgesics. Includes terminal anesthetic surgery, minor
survival surgery with anesthesia, and without significant postoperative pain
(e.g., biopsy) and/or implantation of minor chronic catheters (e.g., femoral
arterial and venous catheters, flow probes, etc.) and/or short-term physical
restraint (<60 minutes) of awake animals and/or induction of minor
behavioral stress.


o                      C: Number of animals:


The research involves procedures
potentially inducing moderate pain, discomfort, or distress which will be
treated with appropriate anesthetics/analgesics and/or chronic maintenance of
animals with a disease/functional deficit. Includes major survival surgery with
anesthesia (e.g., orthopedic surgery on major skeletal components, bowel
resection, cardiac surgery, adrenalectomy) and/or inducement of a functional
deficit (e.g., non-painful tumor inducement, inflammation induced by adjuvants,
inducement of glaucoma, heart failure inducement), and/or physical restraint
(>60 minutes) of awake animals and/or induction of more than minor behavioral
stress.


o                      D: Number of animals:


The research potentially involves
pain, discomfort, or distress (greater than that attending routine injection)
which cannot/will not be alleviated through the administration of appropriate
anesthetic, analgesic or tranquilizer drugs. Examples include: pain research,
radiation testing, toxicity testing and carcinogenesis experiments where
animals become moribund.


In order for a protocol to qualify as
category A, B or C, appropriate anesthetics or analgesics must be used if the
animal will experience more than momentary slight pain. Momentary slight pain
is defined as pain no greater than the level and duration of pain attending a
routine injection. Alternately, the animal must be immediately euthanized upon
evidence of such pain or the protocol is classified as category D.


H.                                                         REGULATORY
QUESTIONS


1.                         THE
ANIMAL WELFARE ACT’S “DUPLICATE EXPERIMENTS” QUESTION


According to the Animal Welfare Act, the ACC is required to determine
whether a reasonable good faith effort was made by the principal investigator
in determining that a proposed experiment is not duplicative. The ACC will be
held responsible if it is subsequently determined that an experiment is
unnecessarily duplicative and that a good faith review of available sources would
have indicated as much.


a.                         Describe
the methods that were used to assure the Animal Care Committee that the animal
activities in this protocol do not unnecessarily duplicate previous
experiments.











b.                         Indicate
resources / scientific literature used:


	
  o

  	
   

  	
  MEDLINE

  	
  (inclusive dates)

  
	
  o

  	
   

  	
  INDEX MEDICUS

  	
  (inclusive dates)

  
	
  o

  	
   

  	
  AGRICOLA

  	
  (inclusive dates)

  
	
  o

  	
   

  	
  EMBASE

  	
  (inclusive dates)

  
	
  o

  	
   

  	
  CURRENT RESEARCH INFORMATION SERVICE

  	
  (inclusive dates)

  
	
  o

  	
   

  	
  Animal Welfare Information Center (see
  appendices)

  
	
  o

  	
   

  	
  Peer review by:

  
	
  o

  	
   

  	
  Consultation
  with an expert in the area of investigation (briefly describe; name,
  position, etc.

  
	
  o

  	
   

  	
  Other:

  
							



c.                         Key words
used in the search for alternatives:


2.                         THE
ANIMAL WELFARE ACT’S “ALTERNATIVE” QUESTION


ALTERNATIVES TO POTENTIALLY PAINFUL PROCEDURES. The use of procedures
potentially causing more than momentary or slight pain or distress to an animal
even though that pain will be relieved by administration of proper drugs,
requires investigators consider alternative procedures and provide a written
description of the methods and sources used to determine that alternatives were
not available and/or were scientifically unacceptable. Note: In designing the
experiment, the investigator should choose procedures that have the least
amount of potential pain, discomfort, distress, or morbidity in consideration
of any limitations imposed by the objectives of the research. Examples include
reduction of the number of required surgeries, less invasive surgery, use of a
less toxic adjuvant, selection of the earliest possible endpoint. This section
should clearly justify why less potentially painful procedures cannot be used
to achieve the specific objectives of the research. This section must also
include a description of the search strategy used to explore alternatives
(e.g., databases consulted, key words used). USDA inspectors require a
description of the key words used in the search of alternatives. Investigators
should be ready to present the results of their searches to the ACC, USDA,  and NIH. USDA inspectors regularly review the
ACC’s records.


a.                         Describe
the methods that were used to assure the ACC that the Principal Investigator
considered alternatives to procedures that may cause more than momentary or
slight pain or distress to the animals. Provide a written narrative description
of the methods and resources.


b.                         Indicate
resources / scientific literature used:


	
  o

  	
   

  	
  MEDLINE

  	
  (inclusive dates)

  
	
  o

  	
   

  	
  INDEX MEDICUS

  	
  (inclusive dates)

  
	
  o

  	
   

  	
  AGRICOLA

  	
  (inclusive dates)

  
	
  o

  	
   

  	
  EMBASE

  	
  (inclusive dates)

  
	
  o

  	
   

  	
  CURRENT RESEARCH INFORMATION SERVICE (inclusive
  dates)

  
	
  o

  	
   

  	
  Animal Welfare Information Center (see
  appendices)

  
	
  o

  	
   

  	
  Peer review by:

  
	
  o

  	
   

  	
  Consultation
  with an expert in the area of investigation (briefly describe; name,
  position, etc.

  
	
  o

  	
   

  	
  Other:

  
						



 















c.                          Keywords
used in the search for alternatives.


I.                            OTHER


1.                         Do any of
the proposed procedures involve the use of Biohazardous materials?


o                      NO - BIOHAZARDOUS materials will not be used in
or disposed of by CAF.


o                      YES - BIOHAZARDOUS materials will be used. (“Application
for the Use of Biohazardous Materials and Animals” must be submitted with the
Animal Use Protocol):


o                      Laser Use (Laser Task Force)


o                      Radioisotopes (Radiation Safety Committee)


o                      Carcinogens, Teratogens, Mutagens, Chemicals


o                      Transmissible/Infectious Agents/Organisms


o                      Recombinant DNA


o                      Other:


2.                         Will
tumor cells, tissue, sera or other biologies from either animal or human
sources be used in or on animal as part of the protocol?


o                       NO


o                       YES:


a.               Please describe
precautions that should be observed with materials or with live/dead animals.


b.              Describe
the tests used to check the tissues and cells for inadvertent contamination by
viruses or mycoplasma.


3.                         ANTIBODY
PRODUCTION


o                      Polyclonal
antibodies:


o                      Monoclonal antibodies:


o                      Adjuvants used.


Adjuvant (If Freund’s
Complete Adjuvant is used, See ACC guidelines)


Route of administration:


Amount injected:


Site preparation:


4.                         TRANSGENIC
MICE


Will
transgenic mice be used?


o                      NO


o                      YES:                                             Source:


Health status: (Describe
the tests used to check the animals for inadvertent contamination by viruses or
mycoplasma.)


5.                         Will
animals be transported to or through patient care areas? (This includes
hallways in Korman Research Pavilion.)


o                       NO


o                       YES:                                          Identify:


Need:











Measures used to protect patients and hospital staff
from possible allergic or emotional
reactions:


Describe route and methods used to move/hide animals
from view:


 


6.                         Will
there be a need to house animals outside of the animal facility for more than
12 hours?


o                       NO


o                       YES                                                Room
Number:                                                          Building:


Please provide justification:


Describe precautions used to prevent contamination of
the area by the animals (e.g., smells, dander) and procedures to assure that
the animals are maintained in a humane manner.


J.                         PERSONNEL
QUALIFICATIONS/INFORMATION


Attach forms:


Principal Investigator Qualifications
in Animal Care and Use

Support Personnel Qualifications in Animal Care and Use


1.                         Occupational
Health


Continuum, Occupational Health Specialists provides occupational health
and workers’ compensation medical services to all
employees at AEHN. If an employee experiences an occupational injury
in the animal facility between the hours of 8:30 AM to 4:30 PM, Monday through
Friday, they are to notify their supervisor or call Occupational Health Office
at 215-456-9061. Arrangements will
be made for the employee be to transported to an Continuum, Employee Health
Office. If an injury occurs after hours they will be treated at the Emergency
Medicine Department. An employee must have an Incident Report with them for
either site. These Incident Reports are available from the CAF office.
Questions regarding this program can be addressed to Ted Feldman, Program
Director for Continuum at 215-533-9424.


Personnel should be instructed to notify their supervisors of potential
or known exposures and of suspected health hazards and illnesses. Employees
experiencing needle sticks or pathogen exposure must seek treatment
immediately.


Personal protective equipment e.g., surgical
masks, exam gloves, isolation gowns, is provided in the animal facility. Also,
along with the animal restrain devices and animal handling gloves, animal
handling videos and training are available through the CAF.


K.                       ASSURANCE
STATEMENTS AND SIGNATURES


“I accept and will conform to all federal and state laws and guidelines
and all institutional policies and procedures concerning the care and use of
animals in research, teaching and testing. I understand that I have a
responsibility to notify the Animal Care Committee in writing of any
substantive changes in the proposed project, changes in personnel assigned to
work under this











application, or in the number of animals or the species to be used,
prior to proceeding with any animal use in such modified projects.


Signature of Principal Investigator and date are required.


The principal investigator ensures that the information provided in the
Animal Use Protocol form accurately reflects the nature of the activities
proposed.











Only one copy of this page
should be submitted to the ACC committee coordinator. This funding source
information should not be included in the copies for distribution to the ACC.


L.                                         FUNDING
SOURCES:


1.                         EXTERNAL
FUNDING


a.                         Please
provide name of the source.


It is important that researchers who are applying for
PHS funds read Pages 17-18, “Vertebrate Animals,” of the grant application
package and complete Section 2F. You may call the ORTD office at 456-7217 if
you have questions or need additional information.


b.                         Grant
Title:


c.                         Amount of
grant or contract:


d.                         Does the
award include administrative


o                       NO


o                       YES, please give dollar amount:


e.                         Does the
agency to which you are applying for fund require an institution approved
animal use protocol?


o                       NO


o                       YES


2.                         INTERNAL
FUNDING


o                       NO


o                       YES:


o            Department:


o            AESG:


o            other


Most, but not all, granting agencies allow to apply for a grant before
the animal use protocol has been approved. If your protocol has been submitted,
but is not yet approved, indicate to the granting agency that the protocol is “pending”.
NIH and NSF normally allow sixty days for approval after the deadline date for
the grant. If no approval from the ACC follows within sixty days of the
deadline date for the grant application, the agency will normally discard your
grant application.


Some funding agencies (such as American Heart Association) require that
the protocol be approved before the grant is submitted. Investigators are
responsible for knowing the requirements of their particular agency.











M.                                      APPENDICES


1.                         Recommended
methods of euthanasia according to species:


	
  Species

  	
   

  	
  Method

  	
   

  	
  Route

  	
   

  	
  Dose

  
	
  Rodents

  	
   

  	
  CO2 asphyxiation

  	
   

  	
  inhalation

  	
   

  	
  until death

  
	
   

  	
   

  	
  cervical dislocation(1)

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
  pentobarbital

  	
   

  	
  IV

  	
   

  	
  150 mg/kg

  
	
   

  	
   

  	
  decapitation(1)

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
  exsanguination(2)

  	
   

  	
   

  	
   

  	
   

  
	
  Rabbits

  	
   

  	
  Euthanasia Solution

  	
   

  	
  IV, IC(3)

  	
   

  	
  0.3 ml/kg

  
	
   

  	
   

  	
  pentobarbital

  	
   

  	
  IV, IC(3)

  	
   

  	
  70-100 mg/kg

  
	
  Cats
  and Dogs

  	
   

  	
  Euthanasia Solution

  	
   

  	
  IV

  	
   

  	
  0.3 ml/kg

  
	
   

  	
   

  	
  pentobarbital

  	
   

  	
  IV

  	
   

  	
  75-100 mg/kg

  
	
  Swine

  	
   

  	
  Euthanasia Solution

  	
   

  	
  IV

  	
   

  	
  0.3 ml/kg

  
	
   

  	
   

  	
  pentobarbital

  	
   

  	
  IV

  	
   

  	
  75-100 mg/kg

  



 






(1) Cervical
dislocation and decapitation without anesthesia is used only when anesthetics
would interfere with experimental results. These techniques can be performed
without anesthesia only when scientifically justified by the PI and approved by
the ACC. Cervical dislocation is limited to mice and small rats weighing less
than 200 grams,


(2) Technique
should only be done in sedated, or anesthetized animals.


(3) Animal must be sedated or tranquilized before IC
procedure.


2.                         Anesthesia
recommendations


	
  Species

  	
   

  	
  Drug

  	
   

  	
  Dose

  	
   

  	
  Route

  
	
  Mouse

  	
   

  	
  ketamine/acepromazine

  	
   

  	
   

  	
   

  	
  IM

  
	
  Rat

  	
   

  	
  ketamine/acepromazine

  	
   

  	
   

  	
   

  	
  IM

  
	
   

  	
   

  	
  ketamine/xylazine

  	
   

  	
   

  	
   

  	
  IM

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  Rabbit

  	
   

  	
  ketamine/xylazine

  	
   

  	
   

  	
   

  	
  IM

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  Pig

  	
   

  	
  Telazol/ketamine/xylazine

  	
   

  	
  2.2 mg/kg of each component

  	
   

  	
  IM

  
	
   

  	
   

  	
  Halothane

  	
   

  	
  to effect (2-3%)

  	
   

  	
  inhaled

  
	
   

  	
   

  	
  Isoflurane

  	
   

  	
  to effect(2)

  	
   

  	
  inhaled

  



 


3.                         Analgesic
recommendations


	
  Species

  	
   

  	
  Drug

  	
   

  	
  Dose

  	
   

  	
  Route

  
	
  Mouse

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  Rat

  	
   

  	
  Butorphanol tartrate

  	
   

  	
  0.04 mg/kg

  	
   

  	
  SQ

  
	
   

  	
   

  	
  Acetaminophen

  	
   

  	
  110-305 mg/kg [2 mg/ml

  	
   

  	
  PO

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  Rabbit

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  
	
  Pig

  	
   

  	
   

  	
   

  	
   

  	
   

  	
   

  



 











4.                         Possible parameters
for evaluation of pain.


·                            Attempting
to protect, move away, or bite


·                            Crying
out when palpated or forced to use affected areas


·                            Licking,
biting, scratching, shaking, or rubbing


·                            Pacing,
lying down and getting up, or shifting weight


·                            Significant
decrease in mobility


·                            Unusual
length of time for recumbency


·                            Reluctance
to move or difficulty in rising


·                            Head
down, tucked abdomen, hunched, facial distortion, or pallor


5.                         Methods
that could be used for monitoring health changes.


·                             Weigh
animals on a regular basis.


·                             Observe
for changes in behavior, activity, or posture.


·                             Observe
for evidence of pain or discomfort in a localized area.


·                             Observe
procedural area for redness, swelling, or discharge.


·                             Observe
for decreased ability to move.


·                             Monitor
food or water consumption


6.                         Endpoint
criteria for removing animals from a study.


·                             Veterinary
opinion


·                             percentage
body weight or loss (example: a loss more than 15% of body weight)


·                             inability
to move


·                             inability
to eat of drink adequately


·                             markedly
reduced response to stimuli


·                             tumor
size


·                             behavioral
abnormalities


·                             signs
of toxicity.


7.                         Animal Welfare Information Center (AWIC) (Your tax dollars at
work.): National Agricultural Library, 10301 Baltimore Blvd., Beltsville, MD
20705, Voice: (301) 504-6212, Fax: (301) 504-5472, Internet:
http://www.nal.usda.gov/awic/. This center can assist investigators in
searching for alternatives and/or perform the search and charge for all online
costs that exceed $25.00.


8.                         Calculation
for the amount of blood you can safely collect from a given animal.


	
  Safe amount of blood volume:

  	
   

  	
  20%

  
	
  Example:

  	
   

  	
   

  
	
   

  	
  4.55 kg

  	
   

  



 











 


	
  weight

  	
  X

  	
  0.06 percent blood volume of body

  
	
   

  	
  0.27 kg

  	
   

  
	
   

  	
  X

  	
  1000.00 mls
  of blood per unit weight

  270.00 mls of blood in the animal

  
	
   

  	
  X

  	
         0.20
  safely take 20% of total blood

  
	
  volume

  	
   

  	
   

  
	
   

  	
   

  	
  54.00 mls can be safely collected

  
				



 











EXHIBIT C

Budget


	
  Principal
  Investigator:

  	
  Heather Kaminski

  
	
  Protocol
  Title:

  	
  Lickometer
  Testing in Mice

  
	
  Company:

  	
  Linguagen

  



 


Research Animals


·                            Linguagen
will purchase all rodents to be used in this protocol.


·                            Linguagen
staff will coordinate animal orders with the CAF staff


·                            Invoices
for animal purchases will be sent directly from the vendors to Linguagen.


Animal Housing and
Care


·                            CAF
will supply standard rodent cages, shelving units, bedding, feed and staff to
provide daily animal care.


·                            CAF
will supply veterinarian services as needed


·                            CAF
staff will prepare (sanitizing and sterilizing) cages for rodent housing.


·                            CAF
will charge $0.79 per cage per day for housed mice.


·                            CAF
will charge $0.97 per cage per day for housed rats.


·                            Estimated
cage count for this budget: 20 to 100 cages per day ($135 to $675 / wk).


·                            CAF
will euthanize and dispose of rodents at Linguagen staff’s request.


Supplies


·                            Linguagen
will provide lab supplies necessary for the procedures described in the
protocol


·                            The
estimate amount is for miscellaneous materials supplied by CAF.


Labor


·                            $12.25
per half hour (non-overtime rate for services other then rodent housing)


·                            The
estimated amount is for possible CAF labor/technician services provided to
Linguagen.


Indirect Costs on the above categories
- 25% (Includes telephone, janitorial service, heating/air conditioning and
electric. All costs associated with the DSL line will be billed directly to
Linguagen.)


Other


·                             Rental
Space


$2.175/month


·                              Cost
per sq. ft. - $25.00 / sq. ft. / year


·                              Total
Space - Approx. l044 sq.ft.


	
  ·                             Lab Space - Room A117
  (Approx. 24’ x 12’)

  	
  288 sq. ft.

  
	
  ·                             Animal
  Housing - Room A114 (Approx. 24’ x 24’)

  	
  576 sq. ft.

  
	
  ·                             Office
  Space - Room A203 (Approx. 10’ x 18’)

  	
  180 sq. ft