Agreement dated March 11, 2004



 EXHIBIT 10.13 
  
 DaVinciTM 
  
 Biomedical research Prod. 
  
 PROTOCOL 
  
 The
Effect of Acidic Fibroblast Growth Factor on the Outcome of 
 Hindlimb Ischemia in the Rat 
  

			
	DaVinci Study Number:	  	DB-106
		
	Sponsor:	  	Cardio Vascular Genetic Engineering
	 	  	 14272 Franklin Ave., Suite 110
 Tustin, CA
92780

		
	Telephone Number:	  	714-368-1520
	Fax Number:	  	714-368-1517
		
	Sponsor Contact:	  	Jack Jacobs
		
	e-mail Address:	  	jjacobs@phagcbiotech.com
		
	Project Site:	  	DaVinci Biomedical Research Products
	 	  	 40 Maple Ave
 South Lancaster, MA
01561

		
	Telephone Number:	  	(978) 368-3477
	Fax Number:	  	(978) 368-3579
		
	Facility Contact Person:	  	Przemyslaw A, Marck
		
	Protocol Issue Date:	  	March 11, 2004

  
 DaVinci Biomedical
Research ProdTM 40 Maple Ave So Lancaster MA 01561
Tel 978-368-3477 Fax 978-368-357 









			
	 DaVinci Biomedical Research ProdTM
 The Effect of Acidic Fibroblast Growth Factor on the Outcome of Hindlimb Ischemia in the Rat
 Study Number: DB-109
	  	03/11/04
Page 2 of 7

  


	1.	OBJECTIVE/LAY SUMMARY: The objective of this study is to evaluate the effect of acidic Fibroblast Growth Factor (FGF) on the outcome of hindlimb ischemia in the rat after 30
days after femoral artery ligation. Hindlimb ischemia will be induced by a ligation and excision of the femoral artery. The limb musculature around the ligation site will be infused with the solution of (FGF) immediately, 2 and 4 days after ischemia
induction. Thirty days after the surgery animals will be sacrificed and limb musculature will be evaluated histologically. Before sacrifice a contrast agent will be delivered to hind limbs (via abdominal artery) and the level of perfusion will be
assessed using fluoroscopy 

  

	2.	REGULATORY REFERENCE: This study will not be performed under Good Laboratory Practice Regulations. 

  

	3.	STUDY DESIGN: 

  

	 	A.	Study Design Table: 

  
 The following diagram presents the study group arrangement: 
  

							
	 Group
Number

	  	 Treatment

	  	 Dose Level

	  	number of
animals


	 1
	  	Sham surgery - FGF	  	DBD	  	12
	 2
	  	Surgery - Vehicle	  	0	  	12
	 3
	  	Surgery - FGF 154	  	DBD	  	12
	 4
	  	Surgery - FGF 154	  	DBD	  	12

  

	 	B.	Rationale for using animals: 

  

	 	a.	Animal Species: (Please explain why this particular species were chosen. and why an animal model is necessary) 

  
 The rat model of hindlimb ischemia is well recognized and widely used.

  

	 	b.	Number of Animals: Forty eight rats will be used. The number of animal selected for this study (12 per group) is necessary to meet the requirements of proper statistical
analysis of results. 

  

	 	c.	Level of Discomfort and/or Pain: Please check one of the following: 

  

	 	 ̈	B. Breeding or Holding Colony Protocols 

  











			
	 DaVinci Biomedical Research ProdTM
 The Effect of Acidic Fibroblast Growth Factor on the Outcome of Hindlimb Ischemia in the Rat
 Study Number: DB-109
	  	03/11/04
Page 3 of 7

  


	 	 ̈	C. Momentary pain and/or distress and no use of pain-relieving drugs, or no pain or distress 

  

	 	x	D. Pain or distress appropriately relieved with anesthetics and/or tranqulizer drugs 

  

	 	 ̈	E. Pain or distress or potential pain or distress that is not relieved with an anesthetics and/or tranqulizer drugs 

  

	 	d.	If C, D&E of section c are chosen have you looked at alternatives and if no alternatives give explanation? (Narrative Description Needed) The Animal Welfare Act and
USDA regulations require that you indicate the information sources you consulted to develop the rationale and methods for this study, to ensure that these studies do not duplicate previous studies, and to ensure that there were no alternatives that
would reduce the use of animals or lessen the amount of pain or distress they might suffer. The USDA requires that you indicate which information sources were searched, when the search was last done, the years covered by the search and what keywords
were used for database searches. Sources might include: the Animal Welfare Information Center, the Current Research Information Service, Medline, Index Medicus, Current Contents, Biosys, specific journals, etc. 

  
 The following literature searches were performed: 
  

							
	 Date of Most
Recent Search


	 	 Years covered
1995-present

	 	 Sources Searched
PubMed

	 	 Keywords Used


	 02/11/04
	 	 	 	 	 	 
	 	 	 	 	 	 	acidic FGF
rat
hindlimb ischemia angiogenesis
FGF I

  
 No alternatives were
found. 
  

	4.	TEST SYSTEM 

  

	 	A.	Animals: 

  
 Common Name: Albino rat 
 Species: Rattus
norvegicus  
 Number of Males: 48 
 Number of Females: 0 
 Age of Onset: on file 
 Weight at Onset: 280-350 g 
  











			
	 DaVinci Biomedical Research ProdTM
 The Effect of Acidic Fibroblast Growth Factor on the Outcome of Hindlimb Ischemia in the Rat
 Study Number: DB-109
	  	03/11/04
Page 4 of 7

  


	 	B.	Source: Animals will be obtained from test facilities certified vendors 

  

	 	C.	Quarantine: All animal will be quarantined upon arrival at the test facility for a period of approximately 7 days. 

  

	 	D.	Randomization: N/A 

  

	 	E.	Identification: Animals will be uniquely identified by tail marks. Individual cage cards will be affixed to each cage and will display the animal number, group number and
study number. 

  

	 	F.	Disposition of Animals: At the end of the study animals will be euthanized with 150 mg/kg pentobarbital sodium (IV). 

  

	5.	ANIMAL CARE PROCEDURE: The procedures described and performed below will be conducted in accordance with the Guide for the Care and Use of Laboratory Animals, USDA APHIS,
Animal Welfare Act and/or in accordance with the Standard Operating Procedures of DaVinci Biomedical Research Prod. 

  

	 	A.	Animal Husbandry and Maintenance: Animals will be individually housed unless group housing is necessary. During the holding and testing period husbandry maintenance and
environmental conditions will be maintained as in the quarantine period, unless changes are needed which will be noted in the protocol. 

  

	 	B.	Environmental Conditions: The conditions in the room will be maintained as follows; room temperature will be maintained 64-84 °F and the humidity will range 30-70%, and
the room will have a 12 hour light/12 hour dark cycle. 

  

	 	C.	Water: Tap water will be provided to the animals ad libitum. Water analysis is performed biannually and will he made available for the study file. Water will be analyzed by
an independent contract facility. The results will include the following: heavy metals, chlorinated hydrocarbons, organophosphates, nitrates, nitrites, total trihalomethanes, dissolved minerals, and a standard plate count. No substances have been
found to date which would be expected to affect the study outcome. 

  

	 	D.	LabChows, Purina Mills, Inc. Rodent Chow Brand will be fed to the animal daily. The lot and expiration will be documented and recorded on file. Chow may differ if specified by
sponsor. 

  











			
	 DaVinci Biomedical Research ProdTM
 The Effect of Acidic Fibroblast Growth Factor on the Outcome of Hindlimb Ischemia in the Rat
 Study Number: DB-109
	  	03/11/04
Page 5 of 7

  


	6.	TECHNICAL PROCEDURES: The procedures described will be performed in accordance with the Standard Operating Procedures of DaVinci Biomedical Research Prod. Data collected will
be documented manually. 

  

	 	A.	Body Weights: Individual body weights will be recorded prior to surgery on the surgery form. 

  

	 	B.	Fasting of Animals: All animals will be fasted at least 12-24 hours prior to surgery. 

  

	 	C.	Preoperative Procedure: 

  
 Anesthesia: Anesthetized will be induced in the chamber filled with isoflurane at 2.5%-4%. After induction anesthesia will be maintained with
isoflurane delivered through the nose cone at 0.5-2.5%. 
  
 Surgical Preparation: The animal hindlimb will be clipped. The operative area will be cleaned with a three alternating scrubs of povidone-iodine scrub and 70% alcohol, once the alternating scrubs are done a final application of
povidone-iodine solution will be applied and allowed to dry (the area will be draped for aseptic surgery). 
  

	 	D.	Surgical Procedure: A vertical longitudinal incision will be performed on one limb. The femoral artery and its branches will be isolated, ligated and excised . The incision
will be closed and animals recovered from anesthesia. Immediately after surgery the area adjacent to the proximal ligation of femoral artery will be injected with a solution of FGF. FGF will be administered again to the same site on day 2 and 4
after surgery. 

  

	 	E.	Postoperative Procedures: 

  

	 	1)	Recovery: The animal will be allowed to recover from anesthesia and transferred to the home cage. 

  

	 	2)	Post-operative Analgesia: Animals will receive Buprcnorphine (0.02 mg/kg, s.c.). if needed. 

  

	 	3)	Post-operative Observations: The animal will be observed after surgery. Skin, hair, respiration, lethargy, fecal output, food consumption, dehiscence of surgical sites,
edema, infection, and vocalization will be observed. All observations will be documented in the study file. If abnormal observations are evident the study director or the attending veterinarian will be notified immediately. 


 











			
	 DaVinci Biomedical Research ProdTM
 The Effect of Acidic Fibroblast Growth Factor on the Outcome of Hindlimb Ischemia in the Rat
 Study Number: DB-109
	  	03/11/04
Page 6 of 7

  


	7.	VETERINARY SUPPORT: The items described below will be performed in accordance to the facilities Standard Operating procedures. 

  

	 	A.	Preventative Therapy: With accordance to accepted veterinary practice the animals may be administered treatment of minor infections, fluid therapy, pain relief, antibiotic
therapy, and any other therapy which may become evident in the course of the animals stay at the facility. All therapy administered to the animals will be documented, and will be made in conjunction with the attending veterinarian and the study
director. The animals will be identified and dose, frequency, and administration will be documented in the study file. 

  

	 	B.	Euthanasia: Animals are to be euthanized at the conclusion, of the project with 150 mg/kg sodium pentobarbital (IV, cephalic vein) in accordance with the American Veterinary
Medical Association (AVMA) Panel of Euthanasia, and Journal of the American Veterinary Medical Association, 218(5): 668-696, 2001. 

  

	8.	PROTOCOL CHANGES: If necessary to change an approved study protocol, changes must be made in writing between the study director and the sponsor. The changes and the
justification for the changes must be included in the section which will be replaced, and the new application must be approved by the study director and the sponsor, of which both parties must sign and date. The amendment will be attached to the
protocol. 

  

	9.	STUDY DEVIATIONS: In the case which a deviation from the protocol or Standard Operating Procedure occurs, the reason for the deviation, the action taken, and the impact on
the study (if any) will be reported by the study director to the sponsor as soon as possible. 

  











			
	 DaVinci Biomedical Research ProdTM
 The Effect of Acidic Fibroblast Growth Factor on the Outcome of Hindlimb Ischemia in the Rat
 Study Number: DB-109
	  	03/11/04
Page 7 of 7

  

 PROTOCOL APPROVAL 
  
 The Effect of Acidic Fibroblast Growth Factor on the Outcome of 
 Hindlimb Ischemia in the Rat 
  
 DaVinci Protocol Number; DB-109 
  

					
	 Sponsor:
	 	 	 	 Cardio Vascular Genetic Engineering,
 14272 Franklin Avc., Suite 110
 Tustin, CA 92780

			
	 /s/ Jack Jacobs
	 	 	 	 3/28/04

	 Signature
	 	 	 	Date
	 Jack Jacobs
	 	 	 	 
	 Sponsor Representative
	 	 	 	 

  

					
	 Facility Performing
 Project:
	 	 	 	 DaVinci Biomedical Research ProdTM
 40 Maple Ave

So. Lancaster, MA 01561

			
	 /s/ Przemyslaw A. Marek
	 	 	 	 3-12-2004

	 Signature
	 	 	 	Date
	 Przemyslaw A. Marek
	 	 	 	 
	 Study Director
	 	 	 	 

  

			
	CHAIRMAN:_______________________________________________________________	 	TODAY’S DATE:
		
	VETERINARIAN:___________________________________________________________	 	TODAY’S DATE:
		
	THIRD COMMITTEE	 	 
	MEMBER:_________________________________________________________________	 	TODAY’S DATE:
		
	 FOURTH COMMITTEE
 MEMBER:_________________________________________________________________
	 	TODAY’S DATE:

  









 PROJECT: 
 GROWTH FACTOR 
 IN HIND LIMB OF RATS 
 (NO. PH1007-Rev-0) 
  
 STUDY SPONSOR 
 PHAGE BIOTECHNOLOGY 
 DR. JACK JACOBS 
 14270 FRANKLIN AVE. SUITE 110 
 TUSTIN, CA 92780 
  
 SPONSOR CONTACT 
 DR. JACK JACOBS 
 PHONE: (714)-368-1520 x15 
 FAX: (714)-368-1517 
  
 PREPARED BY 
 DAVINCI BIOMEDICAL RESEARCH PRODUCTS, INC. 
 PO BOX 1125 
 40 & 20 MAPLE AVENUE 
 SOUTH LANCASTER, MA
01561 
 WEB-SITE: www.davincibiomed.com 
  
 DAVINCI CONTACT 
 MR. JOSEPH VILLANI

 PHONE: (978) 368-3477 
 FAX: (978) 368-3579 
 EMAIL: jvillani@davincibiomed.com 
  
 TODAY’S DATE: 10-21-03 
 DAY ASKED FOR QUOTE: 10-21-03 
 QUOTE 3 PAGES 
 QUOTE GOOD FOR 45 DAYS. 
  









  
 PROJECT 
  
 GROWTH FACTOR 
 IN HIND LIMB OF RATS 
  
 PROJECT COSTS 
  
 USDA
Guidelines and the Animal Welfare Act (Subchapter A, Section 2.31) will be followed. 
  

			
	TIME:	  	To be determined by Joseph Villani of DaVinci & Dr. Jack Jacobs,
		
	COST:	  	 
		
	 	  	 Per USDA Guidelines: Protocol: evaluation, helping client, IACUC evaluation, etc.
 48 CD Rats, 8 wks (24 male, 24 female)
 Filtered boxes and shipping of rats to DaVinci
 Vet visit
 Tech to help vet
 Per Diem for
 48 rats x 14 days acclimation 30 days
2,112

		
	 	  	 Surgery,  1/2 hour per rat x 48 rats, 24 hours total
 24 hrs of
Surgery
 Surgical Specialist 1, 24 hrs
 Surgical Specialist 2, 24
hrs
 Surgical Disposable (Drugs, drapes, etc.), sterile
 Clean
up, 2 hrs/day x 3 days

		
	 	  	 Injection at 0, 2 and 4th day, 48 rats
 2 techs, 5-10 minute per injection, 48 rats, 8 hrs, 3 time points
 48 disposables, 3 times

		
	 	  	 Fluoro Revascularization test
 45
minutes per rat x 48 rats 36 total hours

		
	 	  	 Observation
 Observation, 1 hr/day for
30 days

		
	 	  	 Neeropsy/Tissue Collection, 48 rats @  1/2hr per rat
 21 hrs. of Neeropsy
 Surgical Specialist 1, 24 hrs
 Surgical Specialist 2, 24 hrs
 Neeropsy disposables, non sterile
 Clean up, 2 hrs/day x 3 days
 Carcass Disposal 2 boxes

		
	 	  	 Histology
 Eye(Retina), 5 H&E Slides
x 48 rats
 Kidney, 5 H&E Slides x 48 rats
 Hind limb area
injected, 5 H&E Slides x 48 rats
 Opposite hind limb area not injected, 5 H&E Slides x 48 rats
 Small Histology Report

		
	 	  	Total: $87,023








 TERMS & CONDITIONS 
  

			
	Comments:	  	A change in procedure, quantities, protocol, and time needed, etc. may also change costs. We normally do not change cost for minor changes. They would have to be significant!
		
	Terms:	  	Payment of 50% prior to procedures and 50% upon completion. Past due balances arc subject to a service charge of 1.5% per month.
		
	Cancellation Policy:	  	In the event that the sponsor can not perform the project on the day scheduled and needs to cancel the date, a cancellation fee of $2,000 per scheduled day will be charged to the sponsor
unless cancellation of day is conveyed three weeks in advance. In the event that the project is canceled, all Animals, material purchased for project, labor, etc, will be deducted from the first 50% down payment.

  
 DaVinci Biomedical Research
Products, Inc. 
  

									
					
	Mr. Joseph Villani:	 	/s/ Joseph Villani	 	 	 	 	 	 10-22-2004

	 President
	 	Signature	 	 	 	 	 	 Date

					
	Dr. Jack Jacobs:	 	/s/ Jack Jacobs	 	 	 	 	 	 10/29/03

	 Project MGR
	 	Signature	 	 	 	 	 	 Date