Biomedical subjects

C Allasia

Publications and source records attributed to C Allasia.

34 records · Page 2Linked to original sources

PubMed1995

Digitization of microcalcifications in breast radiographs. Correlation with pathologic data.

A series of 104 nonpalpable breast lesions detected by mammograms and containing microcalcifications were studied. Intraoperative radiographs of intact and sliced specimens were assessed, followed by microscopic diagnosis on frozen sections of areas containing microcalcifications. Microcalcifications detected on mammograms and radiographs of the specimens were digitized and evaluated in accordance with morphometric parameters, including the mean surface, shape factor, bend energy, envelope surface and total surface, total number and concentration of microcalcifications. Benign disorders, atypical hyperplasia and carcinomas accounted for 47.2%, 4.8% and 48% of the tissue lesions, respectively, but the disorders were most often heterogeneous and mixed. Most, but not all, parameters significantly correlated with the three types of radiographs, although radiographs of the sliced specimens provided images of the best quality. Only two parameters, mean size and bend energy, were significantly different (P = .008, .0036) in benign and malignant lesions. It is concluded that image analysis of digitized microcalcifications in radiographs may provide quantitative data helpful in mammogram interpretation.

Breast Neoplasms↗

PubMed1994

Quantitative immunocytochemical assays of P-glycoprotein in breast carcinomas: correlation to messenger RNA expression and to immunohistochemical prognostic indicators.

BACKGROUND: Chemotherapy failure that is due to cellular drug resistance remains a major problem in most cancer patients. One type of drug resistance that has been characterized is the multidrug resistance phenomenon, which demonstrates a reduced ability of cancer cells to accumulate drugs as a result of the effects of an energy-dependent unidirectional drug efflux pump with a broad substrate specificity. This drug pump is composed of a 170-kd transmembrane glycoprotein referred to as the P-glycoprotein (P-gp) that uses energy in the form of adenosine triphosphate to transport drugs through a channel formed by transmembrane segments. PURPOSE: Our purpose was to detect the levels of P-gp expression in frozen untreated breast carcinomas by immunocytochemical assays and to correlate these levels to current prognostic indicators and, in a few cases, to MDR1 (also known as PGY1) mRNA expression by polymerase chain reaction (PCR). METHODS: The immunocytochemical expression of the multidrug resistance gene, P-gp, was investigated using a specific monoclonal antibody (JSB1) against P-gp in 5-microns frozen sequential sections of breast carcinomas obtained from 213 patients. Microscopic images of immunostained preparations were evaluated by image analysis and were compared with MDR1 transcription (mRNA) assessed by PCR in 16 patients. Quantitative P-gp immunocytochemical assays were correlated to histoprognostic factors and immunocytochemical indicators. RESULTS: Among the 213 breast carcinomas tested, 113 (53%) were P-gp positive, but in 28% of the tumors, the immunostained surface accounted for less than 5% of the total area stained. Quantitative immunocytochemistry reflecting the amount of intracellular P-gp antigen strongly correlated (r = 0.865; two-sided, P < .0001; Pearson's test) with the quantitative evaluation of the scanner analysis of mRNA transcripts. The P-gp expression was significantly (two-sided, P < .001) correlated with p53 expression in tumors, to cathepsin D and Ki67 (two-sided, P < .01) immunoreactivity, and to a lesser extent, the detection of estrogen receptor antigenic sites (two-sided, P = .019). P-gp expression was found to be independent of expression of progesterone receptor and pS2, pHER-2/neu, and CD31 in tumors and from patient age, tumor size, histologic types, grades and Nottingham prognostic index, and nodal status. CONCLUSIONS: The quantitative immunocytochemical assays of P-gp are correlated to PCR analysis of MDR1 expression, and such correlations can be useful in evaluating potential multidrug resistance in breast cancer. However, the clinical significance of P-gp immunodetections remains to be further determined.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

PubMed1994

Effects of castration and testosterone on Fel dI production by sebaceous glands of male cats: II--Morphometric assessment.

A morphometric study of cat sebaceous glands was performed to evaluate the effects of castration and testosterone treatments. Skin biopsies were taken in six cats before castration, after castration and after the testosterone injections administered after castration (total number of biopsies: 18). Ninety 8 microns thick sections of each biopsy were assessed for image analysis processing (SAMBA 2005, ALCATEL TITN). The variations in glands and cells size were evaluated on digitized microscopic images by morphometric parameters included in the SAMBA software package. An original software was developed for the analysis of the spacial gland structure. The best morphometric parameters were selected in a first step of the study, and included the nuclear surface (NS), the cell surface (CS) and the nuclear/cellular surface ratio (N/C). These three parameters were then compared in each group of samples for the six cats. It was shown that after castration the N/C (21%) significantly increased compared with prior to castration (12.6%). This 59.8% increase was mainly due to cell cytoplasm shrinking reflecting a decrease of the cell activity. The testosterone administered after castration produced a reverse effect with a N/C ratio back to normal (11.4%) and a significant cell cytoplasm and gland enlargement as shown by the three dimension constructions. This morphometric data correlated with the measurement of sebum and Fel dI productions. The negative effects of castration and the positive effects of testosterone on the sebaceous cells and glands volume favour the hypothesis that cat sebaceous cells are subject to hormonal control this is also likely to apply to the Fel dI production.

Allergens↗

PubMed1993

Cathepsin D immunocytochemical assays in breast carcinomas: image analysis and correlation to prognostic factors.

Immunocytochemical assays of cathepsin D were assessed in a series of breast carcinomas (n = 257) using monoclonal M1G8 anti-total cathepsin D and the avidin-biotin-peroxidase complex. Cathepsin immunoreactivity was compared in frozen and paraffin sections. All tumours were anti-cathepsin-positive. Positive staining was observed in carcinoma and stromal cells and in the extracellular matrix. The amount of immunodetectable cathepsin in tissue was measured by computer-assisted image analysis (SAMBA 2005). Both the percentage of immunostained tumour surface and the mean optical densities were processed as continuous variables for statistical analysis and correlated with prognostic factors. It was shown that cathepsin D was independent of the tumour size, the lymph node status, hormone receptors, and pHER-2/neu overexpression. Cathepsin was significantly correlated with anti-EGFR (P = 0.012) and Ki67 (P = 0.002) immunoreactivity, tumour grade (P = 0.032), vascular invasion (P = 0.0081), proliferation index (P = 0.0045), and, to a lesser extent with AgNORs (P = 0.0504) and the degree of hyperploidy (P = 0.057). Tissue fixation and paraffin embedding significantly decreased cathepsin immunoreactivity. These results show that cathepsin D is not a totally independent prognostic factor in breast carcinomas.

Adult↗

PubMed1993

c-myc protein and Ki-67 antigen immunodetection in patients with uterine cervix neoplasia: correlation of microcytophotometric analysis and histological data.

Cone biopsies were carried out in patients (n = 72) with intraepithelial lesions (CIN 1-3) and squamous microinvasive carcinomas of the uterine cervix. Intraoperative histological examination was achieved in order to obtain small cones with healthy upper resection margins. Also, a tissue fragment was sampled and frozen for the immunocytochemical investigations. Image analysis (SAMBA) of immunoprecipitates was used to evaluate c-myc and Ki-67 antigen expression. Positive c-myc protein staining was observed only on microinvasive carcinomas (70%) and CIN 3 lesions (19%) and was correlated with high Ki-67 immunoreactivity. Ki-67 immunostaining was more extensive: (i) in high-grade than low-grade CIN, (ii) in microinvasive carcinomas than in CIN (P < or = 0.01), (iii) in large than in small lesions. No correlation could be found between the levels of either c-myc protein or Ki-67 antigen expression and either patient age or cytological features of HPV infection.

Adult↗

PubMed1993

Epidermal growth factor receptor in breast cancer: correlation of quantitative immunocytochemical assays to prognostic factors.

Immunocytochemical assays for EGFR were performed on frozen sections from breast carcinomas (n = 209). Results were evaluated by computer assisted image analysis to accurately define the percentage of immunostained surface and the mean optical densities. Thirty seven percent (n = 77/209) of the tumors were EGFR positive, but about one third of them were faintly reactive (35%). No significant relationship was observed between EGFR tumor content and patient age, tumor size, histological type, histoprognostic grade, or axillary lymph node status. A negative correlation was observed with the results of estrogen receptor immunocytochemical assays and a positive correlation with immunodetectable cathepsin D and Ki 67 antigen evaluated according the same method. No correlation was found with HER-2/neu protein, aneuploidy, nucleolar organizor region distribution, and nuclear morphometry, also assessed by image analysis. These results suggest that immunocytochemical assays assessed on frozen sections and evaluated by image analysis are suitable for current and standardized evaluation of EGFR which has been previously documented as a prognostic indicator in breast carcinomas.

Adult↗

PubMed1992

Correlation of nucleolar organizer regions and nuclear morphometry assessed by automatic image analysis in breast cancer with aneuploidy, K167 immunostaining, histopathologic grade and lymph node involvement.

Silver-stained nucleolar organizer regions (AgNORs) in human breast carcinoma were studied using a computer-assisted system of image analysis. Standardized, automatic measurements of 7 morphometric parameters (area, perimeter, shape factor, bend energy, angle, and small and large diameters) performed on paraffin sections and cell imprint were compared and correlated with nuclear morphometry, histopathological grading, tumor growth fraction, (monoclonal Ki67-immunostaining), DNA nuclear content (stoechiometric Feulgen staining) and axillary lymph node invasion. The major findings were as follows: (i) variations in AgNORs and nuclear parameters were correlated, (ii) the ratio of AgNOR area/nuclear area was significantly different in low and high grade tumors, (iii) mean AgNOR parameter values increased significantly with the tumor growth fraction and tumor hyperploidy and were significantly higher in patients with axillary lymph node metastases and (iv) AgNOR evaluation was more accurate for cell preparations than for tissue sections.

Antibodies, Monoclonal↗

PubMed1991

Monoclonal 3C6F9 distribution in human breast carcinomas: image cytometry of immunocytochemical assays.

MoAbF9 immunoreactivity was investigated in frozen sections of 123 breast carcinomas using an avidin or streptavidin biotin peroxidase kit. A standardized computer image analysis system was used to evaluate immunostaining. The percent of cell surface staining and mean optical densities were correlated with morphological criteria of prognosis such as tumor size histological grade, blood and lymph invasion and axillary lymph node involvement, with immunoreactivity to other MoAb, i.e. Ki67, anti-RE and anti-RP, anti-p.HER-2/neu and with tumor aneuploidy and AgNORs content in tumor cell nuclei. Despite some heterogeneity, MoAbF9 was reactive with all breast carcinomas tested. The percent of F9 immunostained cell surface and mean optical density increased with Ki67 immunoreactivity, tumor aneuploidy and AgNORs nucleus surface but were independent of p.HER-2/neu oncoprotein distribution and tumor receptor content. These findings suggest that F9 could not only allow detection axillary lymph node micrometastases but also be used as plasmatic marker for tumor recurrence and metastases.

Antibodies, Monoclonal↗

PubMed1991

Fel d I allergen distribution in cat fur and skin.

Immunohistochemical procedures were performed to ascertain Fel d I antigen (Ag) distribution in cat fur and skin biopsy specimens and to analyze Fel d I allergen concentrations in fur. One hundred strands of fur and 24 skin biopsy specimens (6 by 4 by 3 mm) from shaved areas were collected from 11 different cats. Freshly depilated hairs were immunostained by free-floating monoclonal anti-Fel d I, avidin-biotin-peroxidase complex, and either processed for scanning electron microscopic examination or mounted on glass slides for computer-assisted densitometric analysis (SAMBA system). Skin biopsy specimens were promptly frozen and sectioned just before the immunohistochemical processing. Densitometric analysis of fur demonstrated that immunoprecipitate concentrations were tenfold higher at the root than at the tip. However, this finding may be explained by decrease of the thickness of the hair cortex that varied in similar proportions. The Ag accumulated on the strand surface but may focally penetrate into the medulla through the scale-like cortical interstices. In skin biopsy specimens, Fel d I Ag was found in epithelial squamous cells, within the epidermis and hair follicles, on the surface of the epidermis and hair follicles, and in sebaceous gland cells. These data suggest that Fel d I Ag is produced by sebaceous cells and, to a lesser extent, by basal squamous epithelial cells and that it is stored mainly on the surface of the epidermis and fur.

Allergens↗

PubMed1991

Monoclonal antibody 1BE12 immunoreactivity with human endometrium. Correlations with hormone receptors and proliferation cell markers.

The monoclonal antibody (MAb) 1BE12 has recently been reported to react with several human normal and abnormal tissues. In human endometrium, it reacts more strongly with carcinomas than with normal tissue. To investigate the effectiveness of MAb 1BE12 in identifying cell proliferation in human endometrial cancers, 1BE12 immunocytochemical assays (ICAs) were performed on frozen (n = 47) and paraffin (n = 100) sections with subsequent computer-assisted microcytophotometric (SAMBA) evaluation of immunoprecipitate distribution. MAb 1BE12 immunoreactivity was not impaired by tissue fixation and paraffin embedding. It reacted with normal proliferative endometrium but not with normal secretory endometrium, and immunoreactivity increased with the degree of cell proliferation and malignancy, the amount of immunostaining being greater in invasive carcinomas than in normal proliferative endometrium and endometrial hyperplasia. ICAs showed no correlation between MAb 1BE12 immunoreactivity and estrogen and progesterone receptor antigenic sites. On the other hand, MAb 1BE12 staining in frozen sections increased with Ki67, EGFR, pHER-2/neu, and cathepsin immunostaining. These findings suggest that ICAs on frozen and paraffin-embedded biopsy specimens using MAb 1BE12 along with other markers can be useful for early detection and grading of endometrial carcinoma. The relevance of MAb 1BE12 to the selection of patients for laser ablation of the endometrium rather than hysterectomy is also discussed.

Antibodies, Monoclonal↗

PubMed1990

Human immunodeficiency virus type 1 infection of U937 cells promotes cell differentiation and a new pathway of viral assembly.

The differentiation of U937 monoblastoid cells after human immunodeficiency virus type 1 (HIV-1) infection was studied using the following approaches: reverse transcriptase activity measurement, immunofluorescence labeling, and electron microscopy. For comparison, uninfected U937 cells were induced to differentiate from monocyte to macrophage by phorbol 12-myristate 13-acetate (PMA) or retinoic acid (RA) treatment. Both infected and drug-treated cells showed important and similar ultrastructural cell modifications, with a phenotype that decreased in monocyte specificity and increased in that of macrophages. When U937 cells were induced to differentiate upon HIV-1 infection, a very different pathway of viral production was observed. Production and accumulation of the virus in a vacuolar compartment of intracytoplasmic origin and escape to the antiviral lysosomal activity could explain virus persistence. This makes the cell system a good model with which to study the relationship between HIV-1 production and cell differentiation.

Antigens, CD↗

PubMedDate not supplied

X-raying of sliced surgical specimens during surgery: an improvement of the histological diagnosis of impalpable breast lesions with microcalfications.

Surgical specimens, from 78 patients operated for impalpable breast lesions containing microcalcifications detected on mammographs (breast cancer screening program) were systematically examined by a pathologist during the surgical intervention. The per-operative procedures encompassed (i) X-raying of the unfixed specimens to ascertain that the latter did enclose the screened lesion, (ii) slicing and subsequent X-raying of the specimens slices in order to locate the microcalcifications, (iii) histological evaluation of areas containing the microcalcifications on frozen sections. Thirty-two of the lesions were histologically benign, 55% were malignant and 13% borderline. Forty of the carcinomas were in situ and 60% were invasive. The per-operative histological diagnosis was correct in 65% of the case, erroneous in 10% and uncertain in 25%. Malignancy was never overscored. In 65% of the carcinomas diagnosed during the surgical intervention, the in sano margins excision and axillary lymph node removal could be performed (one stage surgical treatment). These results suggest that X-raying of sliced specimens and histological evaluation during the surgical intervention ascertains that lesions screened by mammographs are effectively and completely removed, and that they are precisely and extensively histologically examined after being identified and located.

Breast Neoplasms↗

PubMedDate not supplied

Inflammatory breast carcinoma: an immunohistochemical study using monoclonal anti-pHER-2/neu, pS2, cathepsin, ER and PR.

Immunocytochemical assays were performed on frozen sections of inflammatory breast carcinomas (n = 22) using the following monoclonal antibodies (MoAb): anti-pHER2/neu, cathepsin, pS2, ER, PR and MoAb Ki67. The distribution of these proteins, known as prognostic indicators, was evaluated with an image analysis system (SAMBA, Alcatel TITN, France). On standard HE stained paraffin sections, only about 50% of inflammatory breast tumors exhibited intradermal tumor cell emboli. All tumors were strongly pHER/2neu positive. All tumors also, but to a lesser degree, were cathepsin and ki67 positive. Conversely, less than 40% were faintly ER, PR and pS2 immunoreactive. The results correlated with the high degree of malignancy of inflammatory breast carcinomas. Therefore the immuno-detection of these markers in addition to standard histological techniques appears to be a useful tool to evaluate the degree of malignancy of breast carcinomas.

Antibodies, Monoclonal↗

PubMedDate not supplied

Monoclonal antibody 83D4 immunoreactivity in human tissues: cellular distribution and microcytophotometric analysis of immunoprecipitates on tissue sections.

Immunocytochemical assays were performed on cell cultures as well as on a wide range of human tissues using a monoclonal antibody, MAb 83D4, produced by a murine hybridoma generated by immunization with a cell suspension from a paraffin block of human breast carcinoma tissue. Frozen breast tissue samples (n = 49) were compared to fixed and paraffin-embedded samples (n = 62). Paraffin sections (n = 194) from a variety of human tissues were compared to breast immunoreactivity. Immunoprecipitates, resulting from positive reactions between 83D4 and Avidin Biotin Peroxidase, were evaluated by computer-assisted microcytophotometry (SAMBA). In some frozen breast samples (n = 27), 83D4 antigen distribution was correlated with tumor cell DNA index, ploidy balance, growth fraction (Ki67), hormone receptor (ER, PR) antigenic sites, NORsAg and oncoprotein pHER-2/neu cell content. MAb 83D4 reacted with 3 breast cancer cells lines (MCF7, T47D and H466B) but not with normal epithelial breast cells in culture. The immunostaining in frozen paraffin sections from breast were similar. Like most normal tissues, normal breast did not react with 83D4. Cellular MAb 83D4 antigen concentration increases with the degree of malignancy but is independent of DNA nuclear content, ER, PR, growth fraction and pHER-2neu in cancers. These results suggested that routine immunohistochemical procedures using MAb 83D4 could facilitate the grading of breast cancer, in particular by allowing detection of microvascular invasions in the breast as well as at a distance and of blood-born micrometastases, especially in bone marrow.

Antibodies, Monoclonal↗

PubMedDate not supplied

Immunodetection of HER-2/neu protein in frozen sections evaluated by image analysis: correlation with overall and disease-free survival in breast carcinomas.

The immunodetection of HER-2/neu oncogene product was performed in 108 breast carcinomas using immunoperoxidase technique. Monoclonal anti HER-2/neu protein was applied on frozen sections. Immunoprecipitates were evaluated by a computerized system of image analysis (SAMBA). The percentage of the immunostained tissue surfaces and mean optical densities were correlated with the 5 year overall and disease-free survival rates. It was shown that in optimal conditions of antigen preservation and standardized method of immunoprecipitates evaluation, 67% of breast carcinomas were more than 20% pHER-2/neu positive. The pHER-2/neu overexpression was not significantly correlated with the overall 5 year survival. However large positive anti pHER-2/neu surfaces were correlated with higher risk of recurrence (p = 0.0013) and of metastases (p = 0.035).

Adult↗

PubMedDate not supplied

Comparison of the prognostic significance of current and modified histological grades in breast carcinomas.

Paraffin sections of 185 breast carcinomas were reexamined in order to compare the current SBR histoprognostic grade (SBR) with to modified methods of grading recently proposed by two groups, Le Doussal et al (MSBR) and Elston et al (SBR-Elston). In each tumor, the SBR, MSBR, SBR-Elston, and each of their components were correlated with recurrence, metastases and survival rates (follow up 7 to 96 months, m = 52, SD = 19) (Kaplan-Meier test). The Nottingham prognostic index (NPI) was computed for each patient, the histological type reevaluated and both were also correlated with the patient follow up. Our results show that the three methods of grading were significantly (p < 0.0001) correlated. The three grades, were significantly correlated with metastases and survival, but not with the recurrence rates. The differentiation, the nuclear pleomorphism and the mitoses count also correlated with the metastases rate and overall survival but the mitoses number appeared to be a stronger prognostic indicator. The MSBR grading made it possible to refine the prognostic of the tumors usually scored as SBR grade 2. The NPI significantly correlated with metastases and survival (p < 0.0001), whereas the histological types were found to have no prognostic significance.

Adult↗