Quantification Of Cell Proliferation Research Articles

Quantification of cell proliferation with the proliferation marker MIB-1 in endomyocardial biopsies after heart transplantation. An aid in diagnosis of graft rejection needing treatment?

The histological grading of endomyocardial biopsies still represents the gold standard in the diagnosis of cardiac allograft rejections. Severe acute rejections (grade 3A/3B, ISHLT) after heart transplantation require immediate high-dose immunosuppressive therapy. The histomorphological differentiation to the rejection requiring no therapy (grade 2) is often difficult. The aim of this study is the improvement of the diagnostic identification of therapy-requiring rejections using quantification of cell proliferation in the inflammatory infiltrate of biopsies. 322 consecutive endomyocardial biopsies from 48 heart allograft recipients were immunohistochemically investigated using the monoclonal antibody MIB-1 binding selectively to the proliferation associated antigen Ki 67. Fifty percent of all biopsies showed cell proliferation: 47.0% in absent or mild rejection (0, 1A, 1B) compared to 88.0% in moderate to severe rejection (2, 3A/3B) (p < 0.01). Proliferating cells were arranged in a focal or perivascular pattern in 66.0% and diffuse in 34.0%. The quantity of proliferating cells per biopsy--but not the pattern--correlated with the grade of rejection: 0.7% in grade 0, 1.4%/5% in 1A/1B, 8.5% in grade 2 up to 18.7% in 3A/3B, and 2.7% in R1 (p < 0.01, ccorr = 0.65). The amount of proliferating mononuclear cells in the inflammatory infiltrate in acute cardiac allograft rejection is proportional to the severity of the rejection and thus an aid in the difficult histological differentiation of grade 3A/3B.

Incorporation of Bromodeoxyuridine as a Method to Quantify Cell Proliferation in Bronchiolar-Alveolar Duct Regions of Asbestos-Exposed Mice

Inhalation of asbestos fibers results in progressive interstitial pulmonary fibrosis in the bronchiolar-alveolar regions of humans and experimental animals. The fibrogenic lesions were described previously by morphometry in a rodent model of asbestos-induced pulmonary fibrosis in our laboratory. Our prior studies also showed that incorporation of tri-tiated thymidine (3H-TdR) into nuclei of epithelial and interstitial cells of bronchiolar-alveolar regions correlates with this histopathologic response. The study presented here describes the early proliferative response in the bronchiolar-alveolar regions in mice after a brief exposure to chrysotile asbestos. Mice exposed to asbestos had significantly increased incorporation of bromodeoxyuridine (BrdU) in interstitial and epithelial cells of first alveolar duct bifurcations and terminal bronchioles at 48 h after exposure. The BrdU incorporation was similar at 24 h after exposure. This study confirms that BrdU provides information on asbestos-induced cell proliferation similar to that of 3H-TdR, but the technology is far safer and less time-consuming.

A cell proliferation assay for small fish and aquatic invertebrates using bath exposure to bromodeoxyuridine

A method is described for bath-labeling small fish, bivalves and crustacea with thymidine analogs to quantify cell proliferation. Medaka ( Oryzias latipes), clams ( Mya arenaria) and copepods ( Pseudodiaptomus coronatus) were exposed to waterborne bromodeoxyuridine (BrdU, 30 mg/l) and fluorodeoxyuridine (FdU, 3 mg/l). A simple spectrophotometric method was developed to monitor uridine uptake from the water by animals. BrdU incorporation into nuclei was detected immunohistochemically, allowing estimation of a proliferation index for different organs and cell types. Bath exposure to BrdU will be useful in study of the response of organisms to chemicals such as epigenetic carcinogens that alter rates of cell proliferation especially in organs with a normally low turnover rate, such as the liver. The procedures may also be applied as an assay for growth, in so far as cell division is central to organism growth.

Cell proliferation after balloon injury of iliac arteries in the cholesterol-fed New Zealand White rabbit.

Acute mechanical injury of an artery results in neointimal hyperplasia that is due at least in part to cell proliferation within the vessel wall. The purpose of this study was to quantify cell proliferation activity in the iliac artery of New Zealand White rabbits after balloon injury and cholesterol feeding. Retrograde pullback balloon injury of iliac arteries was performed, and the animals were then fed a 2% cholesterol diet. At intervals from day 1 through day 35 postinjury, iliac arteries were obtained for histological analysis. Intimal and medial areas were measured morphometrically. Total number of cells within the intima and media was counted. Smooth muscle cell-predominant or macrophage-predominant regions of the intima and media were identified using HHF-35 and RAM-11 immunocytochemical markers, respectively. Number of cells in the proliferative phase of the cell cycle was measured by using the proliferating cell nuclear antigen and bromodeoxyuridine techniques. Thirty-one arteries from 16 rabbits were available for analysis. Total number of cells and number of cells per square millimeter within the media did not change significantly from day 1 through day 35 postinjury. Total number of cells within the intima increased significantly, but the number of cells per square millimeter of intima decreased significantly during the same time period. Proliferative activity was identified in the media between days 3 and 35 with peak activity at day 3 postinjury. Proliferative activity in the intima was present in all specimens from day 8 through day 35. Proliferative activity was present in both HHF-35- and RAM-11-predominant regions of the intima.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of dietary components on the pathobiology of colonic epithelium: possible relationship with colon tumorigenesis.

The concept that diet plays an important role in the initiation and/or development of various types of tumors in man and experimental animals is well documented. The etiology of colon cancer is complex and multifactorial in nature, and there is little information on the dietary components that may act as initiators during colon tumorigenesis. We have evaluated various dietary heterocyclic mutagenic amines present in a typical "Western" diet for their nuclear damaging effect (presumably a genotoxic response) on the colonic epithelium of C57BL/6J mice in vivo. Among the mutagenic amines studied 2-amino-3,4-dimethylimidazo(4,5-f)quinoline and 2-amino-3-methylimidazo(4,5-f)quinoline were very potent inducers of nuclear aberrations. These observations provide us with clues that our daily diet may contain colon-specific genotoxic components. Promotional effects of dietary fat and/or bile acids on colon tumorigenesis have been well studied. Dietary levels of calcium (0.1, 0.5 or 1.0% by weight) appear to modify the toxicity of orally administered fat or cholic acid (assessed by quantifying cell proliferation). The colons of animals consuming 0.1% or 0.5% calcium diet were more susceptible to the toxicity, whereas the colons of those consuming a 1.0% calcium diet appeared more like control colons. These studies demonstrate a profound effect of dietary constituents on the pathobiology of the colonic epithelium which may have a marked influence on the colon tumorigenesis.