Experimental Model of Gene Transfection in Healthy Canine Myocardium: Perspectives of Gene Therapy for Ischemic Heart Disease

OBJECTIVE: To assess the transfection of the gene that encodes green fluorescent protein (GFP) through direct intramyocardial injection. METHODS: The pREGFP plasmid vector was used. The EGFP gene was inserted downstream from the constitutive promoter of the Rous sarcoma virus. Five male dogs were used (mean weight 13.5 kg), in which 0.5 mL of saline solution (n=1) or 0.5 mL of plasmid solution containing 0.5 µg of pREGFP/dog (n=4) were injected into the myocardium of the left ventricular lateral wall. The dogs were euthanized 1 week later, and cardiac biopsies were obtained. RESULTS: Fluorescence microscopy showed differences between the cells transfected and not transfected with pREGFP plasmid. Mild fluorescence was observed in the cardiac fibers that received saline solution; however, the myocardial cells transfected with pREGFP had overt EGFP expression. CONCLUSION: Transfection with the EGFP gene in healthy canine myocardium was effective. The reproduction of this efficacy using vascular endothelial growth factor (VEGF) instead of EGFP aims at developing gene therapy for ischemic heart disease.

Effect of arginine vasopressin on the canine epicardial coronary artery: experiments on V1-receptor-mediated production of nitric oxide

OBJECTIVE: To determine whether arginine vasopressin releases endothelium-derived nitric oxide (EDNO) from the epicardial coronary artery. METHODS: We studied segments of canine left circumflex coronary arteries suspended in organ chambers to measure isometric force. The coronary artery segments were contracted with prostaglandin F2alpha (2 x 10-6M) and exposed to a unique, strong arginine vasopressin concentration (10-6M) or titrated concentrations (10-9 a 10-5 M). RESULTS: The unique dose of arginine vasopressin concentration (10-6M) induced transient, but significant (p<0.05), relaxation in arterial segments with endothelium, and an increase, not significant, in tension in arteries without endothelium. Endothelium-dependent relaxation to arginine vasopressin was inhibited by Ng-monomethyl-L-arginine (L-NMMA, 10-5M) or N G-nitro-L-arginine (L-NOARG) (10-4M), 2 inhibitors of nitric oxide synthesis from L-arginine. Exogenous L-arginine (10-4M), but not D-arginine (10-4M), reversed the inhibitory effect of L-NMMA on vasopressin-mediated vasorelaxation. Endothelium dependent relaxation to vasopressin was also reversibly inhibited by the vasopressin V1-receptor blocker d(CH2)5Try(Me) arginine vasopressin (10-6M) (n=6, P<0.05). CONCLUSION: Vasopressin acts through V1 endothelial receptors to stimulate nitric oxide release from L-arginine.

Endothelium-dependent relaxation in response to poly-L-arginine in canine coronary arteries: implications about hyperpolarization as a mechanism of vasodilatation

OBJECTIVE: To study the mechanism by which poly-L-arginine mediates endothelium-dependent relaxation. METHODS: Vascular segments with and without endothelium were suspended in organ chambers filled with control solution maintained at 37ºC and bubbled with 95% O2 / 5% CO2. Used drugs: indomethacin, acetycholine, EGTA, glybenclamide, ouabain, poly-L-arginine, methylene blue, N G-nitro-L-arginine, and verapamil and N G-monomethyl-L-arginine. Prostaglandin F2á and potassium chloride were used to contract the vascular rings. RESULTS: Poly-L-arginine (10-11 to 10-7 M) induced concentration-dependent relaxation in coronary artery segments with endothelium. The relaxation to poly-L-arginine was attenuated by ouabain, but was unaffected by glybenclamide. L-NOARG and oxyhemoglobin caused attenuation, but did not abolish this relaxation. Also, the relaxations was unaffected by methylene blue, verapamil, or the presence of a calcium-free bathing medium. The endothelium-dependent to poly-L-arginine relaxation was abolished only in vessels contracted with potassium chloride (40 mM) in the presence of L-NOARG and indomethacin. CONCLUSION: These experiments indicate that poly-L-arginine induces relaxation independent of nitric oxide.

Icones plantarum formosanarum nec non et contributiones ad floram formosanam : or, Icones of the plants of Formosa, and materials for a flora of the island, based on a study of the collections of the Botanical survey of the Government of Formosa / By B. Hayata ...

8

Icones plantarum formosanarum nec non et contributiones ad floram formosanam : or, Icones of the plants of Formosa, and materials for a flora of the island, based on a study of the collections of the Botanical survey of the Government of Formosa / By B. Hayata ...

7

Icones plantarum formosanarum nec non et contributiones ad floram formosanam : or, Icones of the plants of Formosa, and materials for a flora of the island, based on a study of the collections of the Botanical survey of the Government of Formosa / By B. Hayata ...

1

Icones plantarum formosanarum nec non et contributiones ad floram formosanam : or, Icones of the plants of Formosa, and materials for a flora of the island, based on a study of the collections of the Botanical survey of the Government of Formosa / By B. Hayata ...

3

Icones plantarum formosanarum nec non et contributiones ad floram formosanam : or, Icones of the plants of Formosa, and materials for a flora of the island, based on a study of the collections of the Botanical survey of the Government of Formosa / By B. Hayata ...

4

Report of the United States Geological Survey of the territories. F.V. Hayden, United States geologist in charge ... [Final reports or monographs. vol. I]-III, V-XIII.

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Icones plantarum formosanarum nec non et contributiones ad floram formosanam : or, Icones of the plants of Formosa, and materials for a flora of the island, based on a study of the collections of the Botanical survey of the Government of Formosa / By B. Hayata ...

6

Icones plantarum formosanarum nec non et contributiones ad floram formosanam : or, Icones of the plants of Formosa, and materials for a flora of the island, based on a study of the collections of the Botanical survey of the Government of Formosa / By B. Hayata ...

10

Icones plantarum formosanarum nec non et contributiones ad floram formosanam : or, Icones of the plants of Formosa, and materials for a flora of the island, based on a study of the collections of the Botanical survey of the Government of Formosa / By B. Hayata ...

9

Icones plantarum formosanarum nec non et contributiones ad floram formosanam : or, Icones of the plants of Formosa, and materials for a flora of the island, based on a study of the collections of the Botanical survey of the Government of Formosa / By B. Hayata ...

5

Icones plantarum formosanarum nec non et contributiones ad floram formosanam : or, Icones of the plants of Formosa, and materials for a flora of the island, based on a study of the collections of the Botanical survey of the Government of Formosa / By B. Hayata ...

2

A biological survey of the waters of Woods Hole and vicinity / By Francis B. Sumner, Raymond C. Osburn, and Leon J. Cole.

Pt.2