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These include histological quantita ti on of intr atumoralvess eldensity and the measurement of angiogenic factors in body Everolimus fluids and tissue extracts. The first demonstration that quantitation of microvessel density might be a reliable predictor of metastasis came fromstud ies on tumor biopsy spec imens of patients with lymph nodenegative breast cancer.Sub sequently, a large number of studies in breast carcinoma and a variety of other tumor types, including lung, prostate, head and neck, rectal, testicular and bladder carcinoma, malignant melanoma, soft tissue tumors, and multiple myeloma, has provided a positive association between tumor angiogenesis and the risk of metastasis, tumor recurrence, or death.However, the inability to demonstrate this association in a number of other studies has led some authors to question the usefulness of a static measure of microvessel density as a prognos tic tool in cancer.One of the requirements for this type of assessment is that immunostained sections must first be scanned at low magnification to select those areas with the greatest numbers of distinctly highlighted microvessels, socalled neovascular hot spots, which almost certainly introduces an element of subjectivity into the measurement.This and other issues related to the measurement process body or lectin markers employed forhighlighting endothelial cells as well as the complexity of the nature of tumor cell growth, invasion, and metastasis formation may explain some of the variability between reported studies.By measuring levels of positive and negative regula tors of angiogenesis in body fluids and tissue extracts, it will be possible in the future to establish an angiogenic profile for patients with cancer and other angiogenesisassociated diseases.To date, virtually all reported studies have been concerned with measuring a single, positive regulator, and most have focused on bFGF, which from a historical point of view antedated VEGF as an Amifostine important positive regulator of angiogenesis.Thus, although bFGF is detectable in trace amounts in the sera and plasma of normal adults, bFGF is increased in maternal and cord betes mellitus.Elevated levels of bFGF have been detected in the urine of patients with a wide spectrum of tumors, including urogenital, breast, and lung, as well as hematological malignancies including lymphoma.High levels of bFGF have also been detected in the cerebrospinal fluid of children with brain tumors.With regard to VEGF, which is detectable in the serum but not the plasma als, elevated levels have been detected in the sera of patients with pulmonary, ovarian, and uterine tumors; in malignant ascites; in cyst fluid and tissue extracts of glioblastomas; in the ocular fluid of patients with diabetic retinopathy and other retinal disorders; in the sera of patients with inflammatory bowel disease; and in the urine of women undergoing gonadotrophin treat ment.In one study, plasma TGFJ levels were found to correlate positively with the extent of tumor vascularity but not with tumor size or underlying liver disease in hepatocel lular carcinoma.Although studies published to date have measured only positive regulators of angiogenesis, it will be important in the future to also consider negative regulators, since their loss may be permissive for allow ing endothelial cells to enter the activation phase of angiogenesis.

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