[CFTR gene variant screening in gamete donation candidates in France: Which indications? How to screen? Why?].

Objectives: According to French recommendations, only the caryotype is carried out as a first line in candidates for gamete donation. The prescription of additional genetic tests for variants responsible for serious monogenic diseases is only recommended in the case of call points. However, cystic fibrosis remains the most common genetic disease with serious consequences in childhood.

[Pre-implantation genetic testing: Comparison between cleavage stage and blastocyst biopsy].

Objectives: Preimplantation genetic testing (PGT) refers to the set of techniques for testing whether embryos obtained through in vitro fertilization have genetic defect. There is a lack of global standardization regarding practices between countries or even from one center to another. In ours, biopsies are preferably performed on day 3 embryos, but also at the blastocyst stage on day 5.

Synthesis and characterization of hydrophobic zeolite for the treatment of hydrocarbon contaminated ground water.

Hydrophobic zeolite was synthesized, modified and characterized for its suitability as a permeable reactive barrier (PRB) material for treatment of hydrocarbons in groundwater. Batch sorption tests were performed along with a number of standard characterization techniques. High and low ionic strength and pH tests were also conducted to determine their impact on hydrocarbon uptake.

Cured meat products without direct addition of nitrate or nitrite: what are the issues?

The growing popularity of food products marketed in the United States as "natural" and "organic" has resulted in a proliferation of marketing efforts to meet consumer demands for these foods. Because natural and organic foods are not permitted to use chemical preservatives, the traditional curing agents used for cured meats, nitrate and/or nitrite, cannot be added to natural and organic processed meat products. However, alternative processes that utilize ingredients with high nitrate content, such as vegetable-based ingredients, and a nitrate-reducing starter culture can produce processed meats with very typical cured meat properties.

Breast cancer chemoprevention phase I evaluation of biomarker modulation by arzoxifene, a third generation selective estrogen receptor modulator.

Purpose: Arzoxifene, a new selective estrogen receptor modulator with strong breast antiestrogen activity and absence of uterine agonist activity, was explored as a potential chemoprevention agent. We performed a multi-institutional evaluation of arzoxifene in women with newly diagnosed ductal carcinoma in situ or T1/T2 invasive cancer.

Experimental Design: In a Phase IA trial, 50 pre- or postmenopausal women were randomized to 10, 20, or 50 mg of arzoxifene daily in the interval between biopsy and re-excision or were enrolled as no-treatment controls.

The efficacy of 9-cis-retinoic acid (aliretinoin) as a chemopreventive agent for cervical dysplasia: results of a randomized double-blind clinical trial.

9-Cis-retinoic acid (aliretinoin) is a pan-retinoid receptor agonist and has been demonstrated in preclinical models to have potent chemoprevention effects. The purpose of this study was to determine the utility of using aliretinoin as a chemoprevention agent in cervical dysplasia. Patients with histological evidence of cervical intraepithelial neoplasia (CIN) 2/3 were randomized in a double-blind manner to receive high-dose aliretinoin (50 mg), low-dose of aliretinoin (25 mg), or placebo daily for 12 weeks.

Her-2/neu gene amplification in ductal carcinoma in situ of the breast.

This study evaluated the relative frequencies of HER-2/neu gene amplification in ductal carcinoma in situ (DCIS)-associated invasive breast cancer and DCIS alone. We examined archival tissue samples of 100 DCIS lesions with an invasive component (cases) and 100 without an invasive component (controls), with cases and controls matched by pathologic nuclear grade. HER-2/neu gene amplification was determined by fluorescence in situ hybridization.

Computer-assisted image analysis-derived intermediate endpoints.

The development of prostatic lesions undergoes a slow progression. To establish efficacy of chemopreventive intervention it is therefore necessary to define surrogate endpoint biomarkers. Such biomarkers should be sensitive in their ability to indicate response.

Chemoprevention with theaflavins of rat esophageal intraepithelial neoplasia quantitatively monitored by image tile analysis.

The objective of the study was to compare three methods of monitoring the inhibition by dietary theaflavins of N-nitrosomethylbenzylamine (NMBA)-induced rat esophageal intraepithelial neoplasia: the mean tile grade, measured by computer-assisted quantitative image tile analysis; tumor multiplicity; and mean tumor size. A "tile" is defined as a small portion of a microscopic image at x 40, 87 x 292 microm in size. The computer divided the image of esophageal intraepithelial neoplasia into a grid of contiguous tiles and measured four tissue features within each tile based on cytonuclear and tissue architectural changes used by pathologists to diagnose intraepithelial neoplasia.

Quantitative grading of rat esophageal carcinogenesis using computer-assisted image tile analysis.

Our objective was to grade, by computer-assisted quantitative image tile analysis, the intraepithelial neoplasia (also called dysplasia) that develops in esophagi of rats given N-nitrosomethybenzylamine (NMBA) for 5 weeks. To perform image tile analysis, the computer divides the video image of the neoplastic epithelium into a row of contiguous small rectangular images, or "tiles," 84 x 292 microm in size, and quantitatively measures four selected tissue features within each image tile. The computer then calculates a tile grade for each image tile as the weighted sum of the four feature measurements, transformed into statistical Z-scores, the weights being determined by Fisher linear discriminant analysis of 300 tile grades of the neoplastic epithelium referenced to the mean tile grade (MTG) of 300 image tiles of normal epithelium.

Image morphometric nuclear grading of intraepithelial neoplastic lesions with applications to cancer chemoprevention trials.

A new image morphometric method of nuclear grading is described and assessed in the context of the evaluation of histological samples from ductal carcinoma in situ of the breast and cervical intraepithelial neoplasia. The method results in a continuous scaled variable, or nuclear grading scale, expressed in SD units from measured normal nuclei from breast or cervix. For a given histological preinvasive neoplastic lesion, the mean nuclear grade of measured nuclei was shown to be analogous to the histopathological nuclear grade of the same lesion assigned subjectively by the pathologist.

Properties of intraepithelial neoplasia relevant to cancer chemoprevention and to the development of surrogate end points for clinical trials.

Cancer chemoprevention is defined as the prevention of cancer by the administration of diet supplements or drugs. A drug discovery effort should therefore focus on finding agents that will avert the process of intraepithelial neoplasia which precedes invasive cancer. Over 30 agents developed by the chemoprevention program at the National Cancer Institute are being tested against intraepithelial neoplasia of many organ sites in more than 80 clinical trials.

Quantitation of preinvasive neoplastic progression in animal models of chemical carcinogenesis.

An assay method that precisely quantitates the cellular and tissue changes associated with early, preinvasive neoplasia is much needed as a surrogate endpoint biomarker (SEB) in clinical trials to predict the potential efficacy of chemopreventive agents in bringing about cancer incidence reduction. Quantification of histological changes at the tissue level are potentially powerful SEB's since these visually apparent changes are common in all neoplastic development, regardless of tissue type or neoplastic cause. Currently, subjective inspection of the histological appearance of sectioned and stained material, or "grading," by experienced pathologists is used to evaluate neoplastic progression.

Properties of intraepithelial neoplasia relevant to the development of cancer chemopreventive agents.

Cancer chemoprevention is concerned with the development of drugs or diet supplements that will avert the onset or stop the progression of the intraepithelial neoplasia which precedes invasive cancer. Two basic processes underlie the onset and development of intraepithelial neoplasia. First is genomic instability (often associated with chronic diffuse epithelial hyperplasia), which is the increased production of genomic structural variants due to unrepaired DNA breaks with secondary formation of abnormal structures, including "mutator" mutations in genes responsible for genomic stability, gene copy amplification or loss from DNA breakage-fusion-anaphase bridge cycles, unequal sister chromatid exchange, and accumulation of double minutes.

Cytology, flow cytometry, image analysis, and interphase cytogenetics by fluorescence in situ hybridization in the diagnosis of transitional cell carcinoma in bladder washes: a comparative study.

The diagnosis of transitional cell carcinoma (TCC) in bladder washes is a diagnostic challenge to cytology. This study assessed the role of flow cytometry (FCM), image analysis (IA), and interphase cytogenetics by fluorescence in situ hybridization (FISH) as adjuncts in the cytodiagnosis of TCC in bladder washes. Forty separate samples of bladder washes were prospectively evaluated by conventional cytology (CY), FCM, IA, and FISH, and the results were compared with the subsequent surgical biopsy specimens which revealed 26 TCC (3 GR I, 6 GR II, 17 GR III) and 14 benign lesions.

Cervical cell recognition and morphometric grading by image analysis.

Cervical cell recognition by morphometric image analysis was compared to human visual cell recognition on the same 6,375 cells from 40 dysplastic, CIS, invasive, and 10 normal pap smears. The experimental approach defined receiver operating characteristic (ROC) curves for morphometric image analysis which could be rigorously compared to previously established human visual cell recognition ROCs on the same cells. Overall performance was measured Az, the area under the ROC curves in the two instances.

A method of correcting DNA ploidy measurements in tissue sections.

A new method of performing DNA ploidy measurements in tissue sections using image cytometry is described. The method involves an image processing "object" filtering operation to remove cut and overlapping nuclei, and DNA correction of larger nuclei for the part of the DNA cut away. The methodology of the technique is developed in detail, and the results of testing using sections of rat liver are presented.

Quality control in image cytometry: DNA ploidy.

DNA ploidy has become a commonly performed quantitative image cytometry test in microscopic pathology. This has led to the need to develop quality control procedures to aid in assuring uniform and reliable test results. There are a number of unique issues related to the emerging technology of image analysis and its routine use as a quantitative microscopic assay that require consideration before establishing a quality control program.

Chromatin texture measurement by Markovian analysis. Use of nuclear models to define and select texture features.

The use of nuclear grade as a prognostic indicator in breast cancer has been limited by its poor interobserver reproducibility. Automated cell classification using digital image analysis is one approach to this problem. Nuclear chromatin distribution, an important feature used in nuclear grading, can be quantitated with texture analysis.

HER-2/neu oncogene expression and proliferation in breast cancers.

Amplification of the HER-2/neu proto-oncogene in breast cancer has been reported to correlate with poor patient prognosis. The proliferation, or growth fraction, of cells has also been shown to be of prognostic importance in breast cancer. A study was conducted to evaluate the correlation between HER-2/neu gene expression and proliferation in breast cancer.

HER-2/neu oncogene expression and DNA ploidy analysis in breast cancer.

This study was performed to evaluate the correlation between HER-2/neu gene expression and DNA ploidy patterns. Forty-five cases of breast-cancer were analyzed. Immunohistochemical staining of HER-2/neu protein on frozen sections was used to detect the HER-2/neu protein, and the Feulgen DNA staining method was used to assess DNA amounts in the same tumor cells.

Comparison of the Papanicolaou and Feulgen staining methods for DNA quantification by image analysis.

The possibility of using archival cytology material to study the evolution of neoplastic disease with regard to DNA content abnormalities was investigated. The accuracy of measuring the integrity optical density (OD) of nuclei that correlates to DNA amounts of those nuclei, on slides stained by the Papanicolaou method, was assessed and compared with a standard Feulgen method. Our data on rat liver nuclei peritoneal washings from patients with ovarian cystadenofibromas and ovarian cystadenocarcinomas suggested that analysis of cytological material using the Papanicolaou method is not reliable and that destaining the slides followed by Feulgen staining provides an optimal and reliable method of DNA quantification.

Biological grading of breast cancer using antibodies to proliferating cells and other markers.

Quantitation of immunohistochemical staining by image analysis was performed on 50 breast cancers stained with the monoclonal antibody Ki-67 to determine the growth fraction and its correlation with tumor grade. A high degree of correlation was shown. For each case the DNA ploidy was determined by quantitation of the DNA Feulgen stain by computerized microdensitometry.

The evaluation of estrogen receptor in primary breast carcinoma by computer-assisted image analysis.

A monoclonal antibody prepared against estrogen receptor has been shown to be specific and sensitive for the detection of estrogen receptor in human breast lesions by use of immunohistochemical methods. Two hundred selected cases of primary breast carcinoma were assayed for estrogen receptor content by biochemical and immunohistochemical procedures. Quantitative evaluation was by biochemical, immunohistochemical, and automated computer-assisted image analysis using the Cell Analysis System's CAS/100 machine (Lombard, IL).

Optical microscope system for standardized cell measurements and analyses.

A fully integrated optical microscope and computer workstation for the pathology laboratory is described along with a system module for that workstation, the Cell Measurement Program (CMP). This module allows for the acquisition and storage of digitized microscope images; measurement of a standard set of cell features, or descriptors, calibrated for accurate densitometry; and a comprehensive set of statistical analyses and display procedures. This system is useful in research in cell biology and in cancer research, allowing the investigator to use the microscope as a measuring instrument.