Microvessel quantification in benign and malignant ovarian tumors

2005 ◽  
Vol 15 (1) ◽  
pp. 58-65
Author(s):  
S. J. Amis ◽  
S. D. Coulter-Smith ◽  
J. C. Crow ◽  
A. B. Maclean ◽  
C. W. Perrett
Keyword(s):  
Correlation Coefficients ◽  
Vessel Density ◽  
Image Analyzer ◽  
Frozen Sections ◽  
Paraffin Sections ◽  
Fixed Tissue ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Microvessel Counts ◽  
Histologic Subtypes

Microvessel density (MVD) in 92 paraffin sections of ovarian samples of different histologic subtypes was correlated with microvessel counts from 58 corresponding frozen sections. Anti-human von Willebrand factor antibody was used as an endothelial marker. MVD was performed in neovascular hotspots using a Quantimet 500+ Image Analyzer. The highest vessel density (HVD) and average vessel density (AVD) of three fields at the × 200 and × 400 magnification were recorded. There was a strong correlation between the HVD and AVD at the × 200 and × 400 magnifications when comparing fixed with frozen sections (correlation coefficients at × 200 for the HVD was 0.37, P = 0.005 and AVD was 0.30, P = 0.02; correlation coefficients at × 400 for the HVD was 0.38, P = 0.003 and AVD was 0.37, P = 0.004). In the fixed tissue, the HVD and AVD at both these magnifications were significantly greater in the group containing functional cysts; this was also the case for the frozen sections. These findings are consistent with the development of a microcirculation necessary for the growth and maturation of such cysts, and this appears to be greater than that in tumors. The good correlation between MVD in fixed and frozen sections suggests that such observations represent a true reflection of ovarian angiogenesis in both physiologic and pathologic states.

Tumor Microvessel Density as a Predictor of Recurrence After Resection of Hepatocellular Carcinoma: A Prospective Study

2002 ◽  
Vol 20 (7) ◽  
pp. 1775-1785 ◽  
Author(s):  
Ronnie Tung-Ping Poon ◽  
Irene Oi-Lin Ng ◽  
Cecilia Lau ◽  
Wun-Ching Yu ◽  
Zhen-Fan Yang ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Microvessel Density ◽  
Disease Free Survival ◽  
Postoperative Recurrence ◽  
Image Analyzer ◽  
Free Survival ◽  
A Prospective Study ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Disease Free

PURPOSE: This study prospectively evaluated the correlation of tumor microvessel density (MVD) with clinicopathologic features and postoperative recurrence in patients undergoing resection of hepatocellular carcinoma (HCC). PATIENTS AND METHODS: Tumor MVD was assessed in 100 patients with resection of HCC using a computer image analyzer after immunostaining for CD34 (MVD-CD34) and von Willebrand factor (MVD-vWF), respectively. Patients were prospectively followed for recurrence. RESULTS: Mean tumor MVD-CD34 (236/0.74 mm2) was higher than mean tumor MVD-vWF (87/0.74 mm2) (P < .001). By multiple regression analysis, tumor size was the only pathologic feature significantly related to tumor MVD-CD34. The median MVD-CD34 was 316/0.74 mm2 in HCCs ≤ 5 cm (n = 46) and 146/0.74 mm2 in HCCs more than 5 cm (n = 54) (P < .001). Among patients with HCCs ≤ 5 cm, those with higher than median MVD-CD34 had worse disease-free survival (at 3 years, 13%) than those with a lower MVD-CD34 (at 3 year, 74%) (P = .002). Multivariate analysis showed that tumor MVD-CD34 was the only significant factor predictive of disease-free survival in patients with HCC ≤ 5 cm. For HCCs more than 5 cm, MVD-CD34 did not have a significant prognostic influence. MVD-vWF did not have a significant prognostic influence on disease-free survival in either HCCs ≤ 5 cm or more than 5 cm. CONCLUSION: This study shows that a high MVD-CD34 was predictive of early postresection recurrence in patients with HCCs ≤ 5 cm and, therefore, may be a novel prognostic marker in this subset of patients.

Angiogenesis as a Prognostic Marker in Early Head and Neck Cancer

1995 ◽  
Vol 104 (9) ◽  
pp. 724-729 ◽  
Author(s):  
Todd G. Dray ◽  
Nicholas J. Hardin ◽  
Robert A. Sofferman
Keyword(s):  
Prostate Cancer ◽  
Head And Neck ◽  
Prognostic Marker ◽  
Head And Neck Carcinoma ◽  
Neck Carcinoma ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Eventual Outcome ◽  
Microvessel Counts

Experimental evidence suggests that tumor growth beyond the earliest stages is dependent on angiogenesis, or neovascularization, and that angiogenesis may also promote metastasis. Recent clinical studies demonstrate that angiogenesis is a prognostic marker in breast, lung, and prostate cancer. To investigate whether tumor angiogenesis also correlates with metastasis and survival in early head and neck carcinoma, we quantified the microvascularity of 106 primary carcinomas prior to treatment and correlated the counts with eventual outcome after 3 to 15 years of follow-up. Microvessels were stained immunocytochemically for von Willebrand factor and then counted by light microscopy. Microvessels were counted per 200x and 400x fields, and their density was graded from 1 to 4, in the area of most intense neovascularization. We found that neither microvessel counts nor density grades correlated with metastatic disease, local recurrence, or survival in early head and neck carcinoma. These results are in contradistinction to those recently reported for other tumor sites.

scholarly journals Control of Autofluorescence of Archival Formaldehyde-fixed, Paraffin-embedded Tissue in Confocal Laser Scanning Microscopy (CLSM)

2001 ◽  
Vol 49 (12) ◽  
pp. 1565-1571 ◽  
Author(s):  
Werner Baschong ◽  
Rosmarie Suetterlin ◽  
R. Hubert Laeng
Keyword(s):  
Bone Marrow ◽  
Sodium Borohydride ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Frozen Sections ◽  
Paraffin Sections ◽  
Fixed Tissue ◽  
Tissue Processing ◽  
Sudan Black B

Confocal laser scanning microscopy (CLSM) offers the advantage of quasi-theoretical resolution due to absence of interference with out-of-focus light. Prerequisites include minimal tissue autofluorescence, either intrinsic or induced by fixation and tissue processing, and minimal background fluorescence due to nonspecific binding of the fluorescent label. To eliminate or reduce autofluorescence, three different reagents, ammonia-ethanol, sodium borohydride, and Sudan Black B were tested on paraffin sections of archival formaldehyde-fixed tissue. Paraffin sections of biopsy specimens of human bone marrow, myocardium, and of bovine cartilage were compared by CLSM at 488-nm, 568-nm and 647-nm wavelengths with bone marrow frozen sections fixed either with formaldehyde or with glutaraldehyde. Autofluorescence of untreated sections related to both the specific type of tissue and to the tissue processing technique, including fixation. The reagents' effects also depended on the type of tissue and technique of tissue processing, including fixation, and so did the efficiency of the reagents tested. Therefore, no general recipe for the control of autofluorescence could be delineated. Ammonia-ethanol proved most efficient in archival bone marrow sections. Sudan Black B performed best on myocardium, and the combination of all three reagents proved most efficient on paraffin sections of cartilage and on frozen sections fixed in formaldehyde or glutaraldehyde. Sodium borohydride was required for the reduction of unwanted fluorescence in glutaraldehyde-fixed tissue. In formaldehyde-fixed tissue, however, sodium borohydride induced brilliant autofluorescence in erythrocytes that otherwise remained inconspicuous. Ammonia-ethanol is believed to reduce autofluorescence by improving the extraction of fluorescent molecules and by inactivating pH-sensitive fluorochromes. The efficiency of borohydride is related to its capacity of reducing aldehyde and keto-groups, thus changing the fluorescence of tissue constituents and especially of glutaraldehyde-derived condensates. Sudan Black B is suggested to mask fluorescent tissue components.

Four agglutination assays evaluated for measurement of von Willebrand factor (ristocetin cofactor activity)

1995 ◽  
Vol 41 (4) ◽  
pp. 510-514 ◽  
Author(s):  
A A Ermens ◽  
P J de Wild ◽  
H L Vader ◽  
F van der Graaf
Keyword(s):  
Von Willebrand Factor ◽  
Correlation Coefficients ◽  
Aggregation Method ◽  
Slide Test ◽  
Cofactor Activity ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Ristocetin Cofactor ◽  
Ristocetin Cofactor Activity ◽  
Better Than

Abstract The concentration of von Willebrand factor (vWf) in patients' plasma can be determined by measuring the ristocetin cofactor activity (vWf R:Co). However, this vWf R:Co assay is time consuming, which limits its routine use. Several commercial vWf R:Co tests, based on agglutination of lyophilized fixed platelets, are available. We evaluated the slide tests and aggregometer assays from Behring and Organon Teknika and compared them with the classic vWf R:Co aggregometer method. The within-run and between-run precisions of the two slide tests were better than those of the aggregometer methods. The correlation studies between the four commercial assays and the classic aggregation method were based on 23 plasma samples (range: 15-450% vWf R:Co). The correlation coefficients, which ranged from 0.923 to 0.950, did not differ significantly (P &gt; 0.1). All four commercial assays gave significantly lower vWf R:Co values than the classic aggregation method (P &lt; 0.01). We conclude that commercially available fixed platelets can be used for the rapid measurement of vWf R:Co with a slide test. The use of the aggregometer is time consuming and may result in a lower precision.

scholarly journals PATIENTS REFERRED FOR BLEEDING SYMPTOMS OF UNKNOWN CAUSE: DOES EVALUATION OF THROMBIN GENERATION CONTRIBUTE TO DIAGNOSIS?

2016 ◽  
Vol 8 ◽  
pp. 2016014 ◽  
Author(s):  
Elena Holm ◽  
Eva Zetterberg ◽  
Susanna Lövdahl ◽  
Erik Berntorp
Keyword(s):  
Thrombin Generation ◽  
Assessment Tool ◽  
Correlation Coefficients ◽  
Von Willebrand Disease ◽  
Assessment Tools ◽  
Clotting Factors ◽  
Viii And Ix ◽  
Bleeding Score ◽  
Von Willebrand ◽  
Willebrand Factor

IntroductionPatients with mild to moderate bleeding symptoms referred for coagulation investigation sometimes never receive a definitive diagnosis. Bleed assessment tools have been developed and validated to assess the severity of symptoms. Global coagulation assays, e.g., the thrombin generation test (thrombogram) have a potential to identify hemostatic defects that are not detected in specific assays.Material and MethodsOne hundred and eighty-five patients referred to our centre because of bleeding symptoms were evaluated using  the bleeding assessment tool (BAT) described by Tosetto and colleagues in 2006.  Blood samples were investigated for thrombin generation (TG) capacity (Technoclone) , in platelet poor (PPP)  plasma , and specific clotting factors, i.e, von Willebrand factor, factor VIII and IX, as well as INR, APTT, platelet count, and platelet adhesion.ResultsOf the 185 patients, five women were diagnosed with mild von Willebrand disease and one male with mild hemophilia A. The remaining 179 subjects (76% females and 24% males with average ages of 33 and 28 years, respectively) were evaluated further. In the total cohort and among women, peak TG, and lag time   correlated with bleeding score (p=0.01 and p=0.04, respectively with correlation coefficients).  No such correlations were found among males. Discussion and conclusion Although our study showed some correlation between TG and bleeding score, results are generally consistent with a previous report which failed to demonstrate the value of TG measurement in a similar setting. In conclusion, the complexity of the mechanisms underlying clinical bleeding complicates the ability to use TG tests as reliable predictors of bleeding. Mild congenital bleeding disorders, especially VWD, should be specifically screened for in patients with mild/moderate symtoms.

Platelet Activation and Aggregation Induced by Recombinant von Willebrand Factors Reproducing Four Type 2B von Willebrand Disease Missense Mutations

1998 ◽  
Vol 79 (01) ◽  
pp. 211-216 ◽  
Author(s):  
Lysiane Hilbert ◽  
Claudine Mazurier ◽  
Christophe de Romeuf
Keyword(s):  
Platelet Aggregation ◽  
Platelet Activation ◽  
Von Willebrand Disease ◽  
Platelet Glycoprotein ◽  
Missense Mutations ◽  
Inhibit Platelet Aggregation ◽  
Wild Type ◽  
A1 Domain ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryType 2B of von Willebrand disease (vWD) refers to qualitative variants with increased affinity of von Willebrand factor (vWF) for platelet glycoprotein Ib (GPIb). All the mutations responsible for type 2B vWD have been located in the A1 domain of vWF. In this study, various recombinant von Willebrand factors (rvWF) reproducing four type 2B vWD missense mutations were compared to wild-type rvWF (WT-rvWF) for their spontaneous binding to platelets and their capacity to induce platelet activation and aggregation. Our data show that the multimeric pattern of each mutated rvWF is similar to that of WT-rvWF but the extent of spontaneous binding and the capacity to induce platelet activation and aggregation are more important for the R543Q and V553M mutations than for the L697V and A698V mutations. Both the binding of mutated rvWFs to platelets and platelet aggregation induced by type 2B rvWFs are inhibited by monoclonal anti-GPIb and anti-vWF antibodies, inhibitors of vWF binding to platelets in the presence of ristocetin, as well as by aurin tricarboxylic acid. On the other hand, EDTA and a monoclonal antibody directed against GPIIb/IIIa only inhibit platelet aggregation. Furthermore, the incubation of type 2B rvWFs with platelets, under stirring conditions, results in the decrease in high molecular weight vWF multimers in solution, the extent of which appears correlated with that of plasma vWF from type 2B vWD patients harboring the corresponding missense mutation. This study supports that the binding of different mutated type 2B vWFs onto platelet GPIb induces various degrees of platelet activation and aggregation and thus suggests that the phenotypic heterogeneity of type 2B vWD may be related to the nature and/or location of the causative point mutation.

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