Characterization by means of Delaunay triangulation and Voronoi paving of the influence of anti-hormone and/or anti-growth factor antibodies on the in vitro cell growth of human colorectal neoplastic cell lines.

A new tool is described which makes it possible to evaluate directly the influence of various growth factors on in vitro neoplastic cell growth on the one hand and to look at a concept of differentiation in terms of population dynamics, on the other. This tool relies upon the digital cell image analyses of Feulgen-stained nuclei and the mathematical method of Voronoi paving. This technique enabled us to characterize the influence on the proliferation and the differentiation of the HCT-15 and LoVo colorectal cell lines of anti-gastrin (G), anti-estradiol (E(2)), anti-epidermal growth factor (EGF), anti-luteinizing hormone-releasing hormone (LHRH), and anti-transforming growth factor alpha (TGF alpha) and beta (TGF beta) antibodies. Two variants were set up with respect to each of the two cell lines, i.e, one growing in culture medium supplemented with 5% fetal calf serum (FCS) and another supplemented with 1% FCS+10 nM G+10 nM E(2). The data show that it is possible to characterize the cell clone structure and to assess growth rate concomitantly by direct cell counts. It further appears that while the anti-hormone and/or anti-growth factor antibody-induced effects on growth were relatively similar, these effects were in sharp contrast at the level of cell clone architecture.

Nuclear DNA content, proliferation index, and nuclear size determination in normal and cirrhotic liver, and in benign and malignant primary and metastatic hepatic tumors.

The distribution values of ploidy, of the proliferation index, of the percentages of diploid cells/case, of nuclear size, and DNA histogram type are described in a series of 92 liver samples from 87 patients. The 92 samples include normal (31 cases) and cirrhotic (14 cases) tissues, benign tumors (7 cases), well-differentiated hepatocellular carcinomas (HCCs, 13 cases), moderately and poorly differentiated HCCs (8 cases), and colorectal glandular metastatic tissues (19 cases). The samples are from either fine-needle aspiration biopsy (FNAB) or histologic imprint smears (HIS). Nuclear assessments were computed on Feulgen-stained nuclei by means of a cell image processor. The results show that the mean DNA index value, the mean nuclear area (NA) value, and the mean percentages of diploid cells per sample are significantly different in the three benign groups under study (normal and cirrhotic tissues and benign tumors) as compared with the mean parameter values from the three neoplastic liver groups (well-differentiated and poorly differentiated HCCs and colorectal metastases). None of these three parameters, however, makes it possible to discriminate clearly between these six histopathologic groups at the individual case level. The mean proliferation index values were significantly lower in the normal tissues and the benign tumors than in the four other histopathologic groups. Recognizing six DNA histogram types, ie, diploid, hyperdiploid, triploid, hypertriploid, tetraploid, and polymorphic, we observed that all the benign samples (the normal and cirrhotic tissues and the benign tumors) exhibited a diploid and/or tetraploid DNA histogram pattern, whereas the neoplastic samples exhibited the six DNA histogram patterns. The combination of the five computerized parameters into a cytologic score (CS) ranging from 5 to 15 permits clear-cut discrimination between nonneoplastic and neoplastic liver cases. The specificity and sensitivity, and the positive and negative predictive values relating to this score were as high as 100%, 95%, 100%, and 96%, respectively.

Ploidy and chromatin pattern analysis as an aid for cervical smear diagnosis.

In the present study we used computer-assisted microscopy to analyze the morphology of Feulgen-stained cell nuclei in cell populations obtained at the same time as routinely performed cervical smears and in the same way. We investigated in a series of 110 cases whether the quantitative morphonuclear description of cytological cervical samples is able to aid pathologists to distinguish between benign and more suspect premalignant lesions. For this task nuclear DNA content, nuclear morphometry (size and anisonucleosis level) and chromatin pattern-related parameters were compiled for each specimen enrolled in the database. A set of 32 normal and 17 high-grade squamous intraepithelial lesion (HSIL) specimens (with diagnostic confirmations) were selected as references and used to establish a discriminant model on the basis of cytometry-generated variables. This model was then used to score the remaining 61 cases in our series (including cases exhibiting benign cellular changes, squamous cells of undetermined significance, low-grade SIL and cancers). The results show that a model discriminating efficiently between normal and HSIL groups can be obtained by combining 5 quantitative features (1 DNA ploidy-related, 2 morphometrical and 2 chromatin texture features). A 97% specificity and an 88% sensitivity characterized the boundary so established. When applied to new cases, the model was in fact able to correct diagnoses for cases which had been down- or up-graded on the basis of the Bethesda system, and provided scores in accordance with histological control.

Multiple clonal chromosome aberrations in a case of childhood focal nodular hyperplasia of the liver.

A case of focal nodular hyperplasia is described that was accompanied by intense reactive stromal changes giving rise to a pseudosarcomatous appearance. Cytogenetic study revealed complex karyotypic abnormalities including five partially identifiable clonal aberrations and one marker chromosome. The composite karyotype was interpreted as: 45-46,XY,add(4)(q21-25)[24], add(11)(p14)[24], add (19)(p13)[15], der(20)t(1;20)(q25;p12)[31], add(21) (q22)[13],-22[3], +mar[2][cp31]. In addition, quadriradial or complex figures, telomeric associations tas, unidentified ring chromosomes, chromosome breaks, and markers were seen in some cells. Such cytogenetic findings, although suggestive of malignancy, could most likely be related to a nonneoplastic condition, i.e., the unusual florid reactive changes associated with this focal nodular hyperplasia.

[Clinical, endoscopic and histologic findings in 1,100 patients of whom 574 were colonized by Campylobacter pylori]. / Données cliniques, endoscopiques et histologiques chez 1100 patients dont 574 colonisés par Campylobacter pylori.

Of 1,100 patients checked by at least two diagnostic tests (urease, histology, culture) 574 (52.1 p. 100) were found to have Campylobacter pylori (C. pylori) in their antral mucosa. Significantly different frequencies of C. pylori (p less than 0.005) were evidenced in the group of patients with active gastroduodenal ulcer (212/298, 71 p. 100), in non-ulcer dyspepsia (NUD) with a previous history of GD ulcer (108/177, 61 p. 100) and NUD without antecedent history of GD ulcer (254/625, 41 p. 100). Whatever the group, males and immigrants were significantly at risk. Chronic alcoholism (greater than 60 g/day) and non-steroid anti-inflammatory drug (NSAID) intake were not predictive for the presence of C. pylori but smokers were significantly at risk when the total (n = 1,100) population was taken into consideration. C. pylori was found in 29 p. 100 of asymptomatic controls (n = 31). There was no significant difference in the frequency and intensity of symptoms when comparing C. pylori+ and C. pylori- patients. The macroscopic aspect of the antral mucosa was not predictive since 51 p. 100 of patients with normal endoscopy were C. pylori+. A strong correlation was observed between the incidence of C. pylori and the severity of gastritis at histology (p less than 0.001) and C. pylori was found in 7 p. 100 of patients with normal histology.(ABSTRACT TRUNCATED AT 250 WORDS)

[Severity of ingestion of caustic substance in children]. / Gravité des ingestions de produits caustiques chez l'enfant.

BACKGROUND: Caustic ingestion is frequent in children, sometimes leading to esophageal stricture. PATIENTS AND METHODS: Between 1988 and 1994, esogastroscopy was performed in 65 children after caustic ingestion. The children were classified in three groups: no lesion (group A), minimal lesions (group B) and severe lesions (group C). Nature of the caustic substance, clinical signs and evolution were compared in the three groups. RESULTS: Median age was 2 years for the 65 children (24 girls, 41 boys). Ingestion occurred at home (94%) during meal periods. Substances were dishwater detergents (n = 14), oven cleaner (n = 10), bleach (n = 9), washing powder (n = 4), others (n = 20), more often in a liquid form (n = 37) than solid (n = 28). Children had no symptoms (57%), presented emesis (n = 20) or abdominal pain (n = 10) not correlated to endoscopic findings, and hematemesis (n = 3) or respiratory distress (n = 4), both symptoms seen only in group C. Buccal lesions (41%) were not correlated to endoscopic findings. After endoscopy, 28 children (43%) were classified into group A and 20 children (31%) in group B. Among the 17 children (26%) of the group C, eight developed an esophageal stricture: seven long strictures requiring replacement of the esophagus, one short stricture requiring repeated dilations. CONCLUSION: Esophageal stricture is still a severe complication after caustic ingestion. These data stress the interest of controlled studies to confirm the preventive role of high dose corticosteroids, and the importance of the prevention of accidental caustic ingestions in children.

Hepatobiliary cystadenoma with mesenchymal stroma: presentation of one case and review of the literature.

We present one case of hepatobiliary cystadenoma with mesenchymal stroma (CMS) which is a very unusual tumor of the biliary tract. These rare neoplasms have been identified as a distinct clinicopathological entity. They occur exclusively in middle-aged women with an average age of 45 years, and are located on the right hepatic lobe. Histologically, CMS are characterized by the presence of a typical dense stroma between an inner epithelial lining and an outer loose connective layer. Malignant transformation may occur and therefore a complete surgical resection of these premalignant neoplasms is indispensable.

Campylobacter pylori-associated gastritis: a double-blind placebo-controlled trial with amoxycillin.

We randomly assigned 45 adult patients with Campylobacter pylori-confirmed antral gastritis to 8 days of treatment with 1 g amoxycillin suspension twice a day, or placebo, according to a double-blind study design. At the end of therapy, 91% of patients treated with amoxycillin demonstrated clearance of the organism from the antrum, compared with 16% in the placebo group (p less than 0.001). Active antral gastritis resolved in 68% of patients in the amoxycillin group versus only 9% in the placebo group (p less than 0.001). No significant difference was observed when looking at the evolution of chronic only gastritis. No significant improvement was observed in the assessment of clinical symptoms and endoscopic appearance. Reappearance of C. pylori and significant worsening of the histological score of active gastritis was observed after 2 wk in all patients. In a second study phase, 18 patients initially not cleared of their bacteria received amoxycillin single blind for 14 days. Clearance of bacteria associated with improvement or resolution of active gastritis was observed in 72% of the cases. In this subgroup, all patients investigated after 1 month were recolonized with C. pylori and, again, had histological active gastritis. We conclude that amoxycillin is effective in treating active antral gastritis associated with C. pylori, but not in preventing relapses, which occur in all cases within 1 month after therapy.

Use of a urea breath test versus invasive methods to determine the prevalence of Helicobacter pylori in Zaire.

The prevalence of Helicobacter pylori infection in Zaire was determined by means of a [14C] urea breath test in 133 asymptomatic subjects, by culture and histological examination of biopsies in 324 consecutive endoscopy patients with chronic epigastric complaints, and by both the breath test and culture/histology in a subset of 92 patients. Sixty healthy Belgian students or hospital laboratory workers were also included for comparison. The prevalence of Helicobacter pylori was significantly higher in asymptomatic Zairian subjects (77.4%) than in the Belgians (30%; p less than 10(-6)). Infection was also acquired much earlier in life in Africans, 66% of the children aged 5 to 9 years already being infected versus none of the Belgian subjects below the age of 20 years. In Zaire, however, the prevalence of Helicobacter pylori in patients with gastroduodenal disorders (87.5%) was similar to that in the group of asymptomatic subjects (77.5%) after adjustment for age and other epidemiological parameters (gender, place of residency, education level, smoking and drinking habits) in a multivariate analysis. The high rate of acquisition of Helicobacter pylori infection in Zaire emphasizes the need to consider the baseline prevalence of Helicobacter pylori in a defined population when studying its association with various diseases.

Effect of antimicrobial therapy on the specific serological response to Helicobacter pylori infection.

The systemic immune response to Helicobacter pylori was studied in 247 infected adult patients before antimicrobial therapy and at different intervals following therapy. Endoscopy with simultaneous collection of biopsies was performed in all patients immediately before treatment, 4 to 6 weeks after the end of therapy and 6 to 12 months later. A 14C-urea breath test was performed 3 to 6 months after the end of treatment. Biopsy specimens were cultured and examined histologically using Giemsa stain. Sera were tested for Helicobacter pylori IgG antibodies with a commercial enzyme immunoassay using species-specific antigens. Overall, Helicobacter pylori was eradicated in 120 patients while the other 127 remained infected with the organism. The follow-up period ranged from 4 weeks to 33 months (mean 10.2 months). Pretreatment IgG levels did not differ significantly between the two groups of patients. Six weeks after the end of treatment a slight but definite decrease in the IgG antibody levels was seen irrespective of treatment success. In the 127 patients who remained Helicobacter pylori-positive, the level of IgG antibodies remained stable or increased with time. A continuous fall in antibody levels was observed following bacterial eradication in the other 120 patients, but the difference in antibody levels between treatment responders and nonresponders became significant only more than six months after the end of treatment (p = 0.001). Serological testing may be useful for monitoring the outcome of long-term treatment of Helicobacter pylori infection and obviate the need for endoscopy.

Breast sensation after superior pedicle versus inferior pedicle mammaplasty: anatomical and histological evaluation.

Despite contradictory information about the course and distribution of the nerves supplying the breast, surgical techniques using an inferior pedicle have been recommended over those using a superior pedicle for preserving the nipple-areolar sensation after surgery. This anatomical study was designed to quantify the nerve branches preserved in inferior and superior pedicles after reduction mammaplasty performed on cadavers. Reduction mammaplasty was done on four fresh cadavers (within 48h of death) using a superior pedicle on the right and an inferior pedicle on the left in a standard way. The pedicle was cut at its base and then fixed in formalin. The base was divided in biopsy specimens and embedded in paraffin. The nerves were quantified and located in each pedicle with haematoxylin-eosin stain and light-microscopic evaluation. Histological evaluation of the pedicles showed the presence of a variable number of nerves (between one and seven) within two superior pedicles and three inferior pedicles. The nerves were located in fibrous tissue and accompanied by vessels in most cases. The nerves were always found superficially and were most likely to be located in the central part of the pedicle. Our results showed that including the nerves within the pedicle is technically uncertain regardless of the mammaplasty technique used. The final recovery of sensation in the breast after mammaplasty seems to result from the regeneration of severed cutaneous nerve branches or the remaining cutaneous innervation rather than the preserved adjacent cutaneous branches.

[Primary sclerosing cholangitis: contribution of retrograde cholangiography in follow-up: apropos of 15 case reports]. / Cholangite sclérosante primitive: contribution de la cholangiographie rétrograde dans le follow-up: à propos de 15 observations.

From 1975 to 1989, primary sclerosing cholangitis (PSC) was diagnosed in 15 patients especially based on cholangiographic features associated with clinical, biological and histological findings. 93 p. cent of patients had altered liver enzymes at the diagnosis time but only 26% was asymptomatic. 33 p. cent had a concomittant inflammatory bowel disease. Retrograde cholangiography showed injuries of the intrahepatic ducts in 93 p. cent; the more frequently observed lesions were short and multifocal strictures, associated with irregularities of the biliary wall or decreased arborization of intrahepatic bile ducts, but without marked dilatation. Mean follow-up was 44 months (3 to 120). Actuarial survival was 48 p. cent at 5 years. In 4 patients, because of poor clinical and biological (serum bilirubin greater than or equal to 4 mg/dl) evolution, a second retrograde cholangiography was performed showing in all cases apparition of marked ductal dilatation in extra-hepatic (one case) or in intra-hepatic bile ducts (three cases). We diagnosed one benign extra-hepatic stricture, two cholangiocarcinoma and one biliary cirrhosis complicating PSC. We concluded that retrograde cholangiography is necessary if jaundice appears to diagnose every complications of PSC. Apparition of marked ductal dilatation should be the witness of pejorative evolution, especially apparition of cholangiocarcinoma that must be excluded.

Multiparameter digital image analysis of biliary, ampullary, and pancreatic adenocarcinomas.

Periampullary adenocarcinomas arise from pancreatic, biliary, or ampullary ductal epithelium. Their origin is difficult or impossible to discern by routine light microscopy of cytologic or small biopsy specimens. We used image analysis to describe the morphologic features of Feulgen-stained nuclei of biliary and ampullary adenocarcinomas and to compare these findings with those of pancreatic ductal adenocarcinomas. Surgically resected cases of ampullary adenocarcinoma (n = 7) and cholangiocarcinoma (n = 26) were selected on the basis of available formalin-fixed, paraffin-embedded tissue and diagnostic clinical data. Disaggregated nuclei were stained by the Feulgen reaction and analyzed using a digital image processor. Data from 15 morphonuclear parameters were assessed and compared with the results from the morphonuclear analysis of 22 pancreatic ductal adenocarcinomas. Multivariate analysis with canonical transformation of the data defined the ampullary adenocarcinomas and cholangiocarcinomas as occupying similar factorial distributions, whereas the pancreatic carcinomas were separate and distinct. Monovariate analysis identified seven parameters distinguishing pancreatic carcinoma from both ampullary carcinoma and cholangiocarcinoma, with P values < or = 0.001 and two others having P values < or = 0.01. Adenocarcinomas of ampullary or bile duct origin possess similar morphonuclear features described by image analysis. Image analysis provides a mechanism to discriminate adenocarcinomas arising from the bile ducts or ampulla from those arising in the pancreas.

Chemotherapy-induced nuclear alterations of morphologic and genomic characteristics in a human colon cancer grafted onto nude mice.

PURPOSE: A human Dukes B colonic adenocarcinoma was grafted onto 40 nude mice. The mice were divided into four groups, one control and three representing experimental conditions. Animals in the three experimental groups received either adriamycin (ADR), 5-fluorouracil (5-FU), or camptothecin (CPT) over a 25-day period beginning 34 days after grafting. Control animals received saline on an identical schedule. Animals were killed 105 days after grafting. METHODS: The effect of therapy was assessed by three techniques: 1) tumor size was periodically measured during the life of the animals, 2) modifications of APC, Ki-ras, and p53 genes were studied by polymerase chain reaction, dot-blot analysis, restriction analysis, and DNA sequencing, and 3) image cytometry of Feulgen-stained material was used to characterize 15 parameters describing morphometric, densitometric, and textural features of tumor nuclei. RESULTS: When compared with controls, tumor growth (size) was maximally suppressed by treatment with CPT (P < or = 0.001). Growth was inhibited significantly by treatment with 5-FU (P < or = 0.01); no statistical difference in tumor size was observed between controls and animals treated with ADR. Modifications of APC, Ki-ras, and p53 genes were not observed; however, treatment did inhibit amplification of APC and p53 genes. CONCLUSIONS: The 15 morphonuclear parameters were assessed to define populations of cell nuclei altered by chemotherapy. Although CPT maximally suppressed growth, it did not alter nuclear morphology when compared with controls. Treatment with either 5-FU or ADR resulted in nuclear morphologic alterations defined as distinct populations using multivariate analysis. Nonsupervised linear discriminant analysis was used to quantify the relative proportions of these populations. Four morphonuclear parameters were identified, which discriminated nuclei exposed to either ADR or 5-FU from controls.

Heterogeneity of DNA ploidy, proliferation index and nuclear size in human colorectal carcinomas.

Measurements of DNA ploidy, proliferation index and nuclear area were performed on 210 samples taken from 15 human colorectal tissues. The tissues were divided into four groups labeled G1, G2, G3 and C. For each of the 15 tissues 9 samples were taken from the so-called unaffected--i.e., marginal--mucosa (G1-G3 groups) and 5 from the tumor (C group). The 9 samples from the unaffected mucosa of each tumor were obtained at a distance of 10 cm (3 samples/tissue, G1 group), 5 cm (3 samples/tissue, G2 group) and 1 cm (3 samples/tissue, G3 group) from the tumor. Computerized cell image analysis was carried out on Feulgen-stained cell suspensions obtained from paraffin-embedded, formalin-fixed tissues. The results revealed that four to five analyses are necessary to detect minor aneuploid cell nuclei populations in human colorectal tumors. A definite homogeneous diploid pattern was found in the G1-G3 samples. In contrast, proliferative activity varied widely between the normal and tumor samples, with such variations observed at both the sample-to-sample and tissue-to-tissue level. The nuclear area also varied markedly across the samples from a given tissue--i.e., both marginal and tumoral and across the tissues themselves. Finally, we observed that the diploid tumors, the nuclear sizes of which varied as widely as those of the aneuploid tumors, possessed a higher proportion of highly proliferating samples than did the aneuploid.

Severe hepatotoxicity related to benzarone: a report of three cases with two fatalities.

We report three cases of severe hepatotoxicity related to benzarone, a benzofuran derivative. Our cases include a 35-year-old woman with (sub)fulminant hepatitis, a 67-year-old woman with macronodular cirrhosis, and a 68-year-old man with severe chronic active hepatitis and cirrhosis, with positivity of anti-smooth muscle antibodies. Two patients died. We stress the potential of benzarone to cause hepatotoxicity, which usually resembles severe chronic active hepatitis. Our cases constitute the most severe cases of benzarone hepatotoxicity reported so far, and comprise the first cases of (sub)fulminant hepatitis and cirrhosis related to benzarone.