The role of vascular endothelial growth factor and its receptor Flk-1/KDR in promoting tumour angiogenesis in feline and canine mammary carcinomas: a preliminary study of autocrine and paracrine loops.

Vascular Endothelial Growth Factor (VEGF) and its receptor KDR are involved in the regulation of angiogenesis and are up-regulated in a number of tumours in humans and in particular, breast cancer. We therefore evaluated the prognostic potential of the angiogenetic process in feline and canine mammary carcinomas by the immunohistochemical assessment of VEGF expression and micro vessel density (MVD) quantification and examined the interplay between VEGF and KDR. These variables were related to some relevant clinicopathological parameters and to overall survival (OS). VEGF and KDR expression were evaluated in epithelial, stromal and endothelial compartments in order to identify autocrine and/or paracrine loops. In dogs an increased VEGF expression did not show any statistical correlation with the clinicopathological parameters examined and was not correlated to a poorer prognosis. MVD was found to be significantly correlated to the histologic type (P=0.04), tumour grading (P=0.02), and to the OS (P=0.01). In cats VEGF expression was significantly correlated to tumor grading (P=0.01) and OS (P=0.03), while no significant associations were found between MVD and the other parameters. VEGF and KDR were found to be detected on the epithelial, and/or endothelial and/or stromal cells of the carcinomas in both species, suggesting indications for some possible autocrine and paracrine loops. Our results encourage further studies on the possible prognostic role of VEGF and MVD in canine and feline mammary tumours and on the role of growth factors and their receptors in promoting tumour proliferation and an "angiogenetic shift". The VEGF/KDR system may play a role in malignant transformation and tumor progression.

Vascular tissue banking: state of the art.

INTRODUCTION: The cardiovascular homograft banks in Italy were set up in 1994 in Milan (Lombardia) and Treviso (Veneto) and in 2001 in Bologna, Emilia Romagna. In this study we briefly summarize the data from Emilia Romagna Cardiovascular Tissue Bank. MATERIAL AND METHODS: In Emilia Romagna, vascular homografts were harvested from brain-dead multiorgan donors (aged 15-55 years) by a dedicated vascular surgery team. All donors were virologically screened for human immunodeficiency virus (HIV), hepatitis B and C, Treponema pallidum, cytomegalovirus (CMV), and Toxoplasma. After transferring the vascular homografts to Emilia Romagna Cardiovascular Tissue Bank facilities, the arteries were prepared, classified (class III to I), and transferred to an antibiotic-containing solution under a laminar flow cabinet. After the decontamination, all homografts were cryopreserved and stored in the vapour phase of liquid nitrogen. Microbiological tests were performed in all phases of preparation. Samples were routinely taken from 1 vessel and formalin fixed for the histology. Bags with cryopreserved homografts were sent in dry ice to the hospitals when required and thawing protocol of the Bank was included. RESULTS AND CONCLUSIONS: From January 2002 to October 2004, 543 homografts from 125 heart-beating donors were harvested and transferred to Emilia Romagna Cardiovascular Tissue Bank. After preparation, 459 of 543 (85%) were cryopreserved and stored. Vascular homografts classified class I were discarded. Other criteria of rejection were: (1) positive serology, and (2) persistent positive microbiology after decontamination. From March 2002, 333 cryopreserved homografts were assigned to several vascular surgery departments in Italy. The assessment of 3-year activity of Emilia Romagna Cardiovascular Tissue Bank might be used as an indicator of the efficiency of selecting, cryopreserving, and allocating quality-controlled vascular homografts.

Smooth muscle cell injury after cryopreservation of human thoracic aortas.

The cryopreservation protocol we use for arterial reconstructive surgery has been studied to evaluate smooth muscle cell (SMC) structural integrity and viability before implantation. Samples of human thoracic aortas (HTA) were harvested from five multi-organ donors. Sampling included unfrozen and cryopreserved specimens. Cryopreservation was performed using RPMI with human albumin and 10% Me(2)SO in a controlled-rate freezing apparatus. Thawing was accomplished by submerging bags in a water bath (39 degrees C) followed by washings in cooled saline. In situ cell preservation as investigated by light and transmission electron microscopy showed that SMCs from cryopreserved HTA had nuclear and cytoplasmic changes. A TUNEL assay, performed to detect DNA fragmentation in situ, showed increased SMC nuclear positivity in cryopreserved HTA when compared to unfrozen samples. 7-AAD flow cytometry assay of cells derived from cryopreserved HTA showed that an average of 49+/-16% cells were unlabeled after cryopreservation. Organ cultures aimed to study cell ability to recover cryopreservation damage showed a decreasing number of SMCs from day 4 to day 15 in cryopreserved HTA. In conclusion, the cryopreservation protocol applied in this study induces irreversible damage of a significant fraction of arterial SMCs.