[Bladder located gastric heterotopy: a case report confronting embryology to histopathology]. / Hétérotopie gastrique de siège vésical: un cas clinique au carrefour de l'embryologie et de l'histopathologie.

We report a case of bladder located gastric heterotopy, which has never been described, to our mind in the scientific literature. We discuss the diagnosis and the physiopathological mechanisms that may have been involved in the genesis of such a lesion.

Expression of REG protein during cell growth and differentiation of two human colon carcinoma cell lines.

We localized REG protein in Paneth cells and nonmature columnar cells of the human small intestinal crypts and speculated that this protein was associated with growth and/or differentiation. The aim of this study was to determine whether REG protein is present in two human colon cancer cell lines that exhibit enterocytic differentiation after confluence and to investigate changes in the level of its expression during growth and differentiation. Results were compared to those obtained on cells that remain undifferentiated. Western immunoblotting and immunofluorescence demonstrated the presence of REG protein in the three cell lines. With the antisera against human REG protein, the staining was diffusely spread throughout the cytoplasm at Day 2, and after Days 3-4 it appeared to have migrated to cell boundaries. After confluence, we observed only a punctate staining array along cell boundaries, which disappeared at Day 15. REG mRNA expression was demonstrated by RTPCR and REG mRNA hybridization until Day 13, but not after, in the three cell types. REG protein may be involved in cellular junctions. Its presence appears to be associated with the cell growth period and the protein must be downregulated when growth is achieved and differentiation is induced.

Pancreatic trypsinogen I expression during cell growth and differentiation of two human colon carcinoma cells.

Pancreatic trypsin has been found to induce tight junction or dome formation in some colon cancer cell lines (HT-29, Caco-2), and a tumor-associated trypsinogen, trypsinogen type II, has been isolated from another colon cancer cell line (COLO 205). We have tried to determine if trypsinogen is present and how its expression varies during cell culture in HT-29 Glc+/- and Caco-2 cells, which exhibit enterocytic differentiation, and in HT-29 Glc+ cells, which never differentiate. Trypsinogen mRNA presence and expression were demonstrated in these cells by mRNA hybridization, RT-PCR, cytoimmunofluorescence, Western immunoblot analysis, and gel filtration. Trypsinogen was found to be trypsinogen type I and was mainly in zymogen form in culture media. Differentiating cells exhibited variations in trypsinogen I expression, but cells that remained undifferentiated did not. In the differentiated cells, a high and transient peak in trypsinogen I expression was observed during the first steps of differentiation.

Molecular characterization of the gene encoding the DNA gyrase A subunit of Streptococcus pneumoniae.

The gene encoding the DNA gyrase A subunit of Streptococcus pneumoniae was cloned and sequenced. The gyrA gene codes for a protein of 822 amino acids homologous to the gyrase A subunit of eubacteria. Translation of the gene in an Escherichia coli expression system revealed a 92-kDa polypeptide. A sequence-directed DNA curvature was identified in the promoter region of gyrA. The bend center was mapped and located between the -35 and -10 regions of the promoter. Primer extension analysis showed that gyrA transcription initiates 6 bp downstream of an extended -10 promoter. The possible implications of the bent DNA region as a regulatory element in the transcription of gyrA are discussed.

Modulation of proliferation, second messenger levels, and morphotype expression of the rat intestinal epithelial cell line IEC-6 by fermented milk.

Trophic effects of milk fermented with Lactobacillus helveticus, Lactobacillus paracasei ssp. paracasei, Bifidobacterium sp., or the combination of Lactobacillus bulgaricus and Streptococcus thermophilus (yogurt) were studied on the IEC-6 intestinal epithelial cell line. Incorporation of [methyl-3H]thymidine, mitochondrial dehydrogenase activities, cyclic AMP production, and differentiation of levels of the IEC-6 strain were evaluated between the 15th and 30th passage in culture. All fermented and unfermented milks enhanced trophic responses of IEC-6 cells in a dose-dependent manner. Compared with the corresponding milks, supernatant fractions were more effective in stimulating mitochondrial dehydrogenase response. Fermented milk supernatants were also more effective than the corresponding unfermented fractions. Increases in DNA synthesis and cyclic AMP confirmed the activation observed with mitochondrial dehydrogenase. Yogurt induced the more trophic response with an increased number of the more differentiated cell morphotype. Fermentation with L. casei also demonstrated an important trophic adaptation of IEC-6 cells. Milk processing by lactic acid bacteria enhanced trophic and proliferation responses of intestinal epithelial cell line IEC-6. These results suggested that IEC-6 cells could represent an accurate and easy in vitro model for testing the trophic quality of various nutrients and for an optimization of physiological digestive functions.

Immunohistological localization of regenerating protein in ocular structures.

PURPOSE: To investigate the presence of pancreatic stone protein (PSP)/reg (regenerating) protein in eyes and extraocular structures of rabbits, monkeys and man, according to species and age. METHODS: Rabbit eyes, with normal or de-epithelialized corneas, were taken from albino and pigmented animals. Monkey eyes were taken from cynomolgus monkeys, 2 years old. Stillborn and adult human eyes were obtained after autopsy. They were studied by immunohistochemistry methods, using a monoclonal antibody raised against human PSP. RESULTS: On rabbit ocular structures, the anti-PSP monoclonal antibody showed a strong reactivity at the level of basement membranes and basal poles of the cells of corneal and conjunctival epithelia and on basement membranes of skin and palpebral conjunctiva in eyelids. On de-epithelialized rabbit eyes, the remaining basement membrane was labeled while, along the re-epithelialization, the anti-PSP reactivity appeared with the migrating cells which cover the denuded cornea. On young monkeys, the whole corneal epithelium was reactive. Similar results were obtained from stillborn eyes, whereas no reactivity was detected on autopsy specimens from aged persons. CONCLUSIONS: As in other tissues and organs, the reg protein, in the eye, is found in structures known for their continuous and rapid renewal. This protein seems not to exist (or persist) in eyes from aged donors while it is strongly expressed in young donor eyes (monkey, stillborn baby) as well as in regenerating corneal epithelium. These findings enforce the hypothesis about the involvement of reg protein in cell proliferation and differentiation phenomena and its probable correlation with aging.

[Isolated smooth muscle cells of the human colon. Cytophysiological study]. / Cellules musculaires lisses isolées du côlon humain. Etude cytophysiologique.

Gross normal specimens of human distal colon were obtained at operation for cancer. Smooth muscle cells were separated from internal and external layers of the muscularis. They were dissociated by digestion with collagenase, isolated and concentrated by successive centrifugations. Colonic smooth muscle cell contraction was measured using various concentrations of carbamylcholine (10(-9) to 10(-4) M); relaxation was tested using atropine (10(-9) to 10(-4) M) on colonic smooth muscle cells pre-contracted by carbamylcholine. Compared with previous descriptions, human smooth muscle cells were smaller than in other species with an enlarged distribution of cell size (30 microns to 150 microns in length). Significant dose-response curves were obtained for both carbamylcholine and atropine. However, 3 original points characterized human colonic smooth muscle cells: a) the cells isolated from the internal layer were significantly more sensitive than those isolated from the external layer (10(-9) M vs 10(-7) M); b) for the muscle cells isolated from both the internal and external layers, small colonic smooth muscle cells were significantly more sensitive. On the other hand, these cells were shown to be located near conjunctive septae, and intramural plexuses; c) analysis of contraction curves demonstrated a more efficient response for colonic smooth muscle cells of the internal layer than for those of the external layer of the muscularis.

Immunocytochemical demonstration of a pancreatic secretory protein of unknown function in human duodenum.

We have previously isolated from human pancreatic juice a secretory glycoprotein of 19 KD (P19), devoid of known enzymatic activity. P19 gave by proteolysis a protein of 14 KD (P14), at first named protein X and also called pancreatic thread protein or pancreatic stone protein. Specific rabbit immunosera prepared against P19 and P14 were applied to localize these proteins in human small intestine. By comparison, antibodies directed against some human pancreatic enzymes (amylase, lipase, chymotrypsin, trypsinogen 1, trypsinogen 2, and trypsin 1) were also tested. Positive immunoreactivity was observed on Paneth cells with antisera directed against trypsinogens, trypsin 1, and P19-related proteins. In addition, antisera directed against P19-related proteins stained the columnar cells located in the crypts of Lieberkühn. These original findings are a further indication of the resemblance between Paneth and pancreatic acinar cells but show that their functional analogy is only partial. On the other hand, the presence of P19-related proteins on non-mature columnar cells suggests that this differential distribution is a consequence of differentiation.

Silent human pancreatic glucagonoma and "A" nesidioblastosis.

Surgical fragments of healthy and tumor-bearing pancreas from a patient with pancreatic tumor were studied by electron or light microscopy, histochemistry, and immunocytochemistry (human insulin, glucagon, somatostatin, gastrin, and bovine pancreatic polypeptide). Histological results were compared to those obtained by radioimmunoassay, both in tumor and serum. The tumor was identified as a glucagonoma because reactions for Grimelius' silver impregnation and immunoreaction with an antiserum against glucagon were positive and because a very high level of glucagon in the tumor was observed. Insulin, somatostatin, and gastrin levels remained normal, both in tumor and serum, but the glucagon level was normal in serum. Associated with this silent glucagonoma, an uncommon nesidioblastosis was also diagnosed with many A cells irregularly mixed with acinar cells, isolated or clustered in small groups. Acinar "intermediate" cells of "A" type were also observed. Such associative histopathological processes evoked possible development of an endocrine tumor from nesidioblastic-like tissue. Its embryogenic origin remained uncertain.

[Yag laser photocoagulation and monopolar electrohydrocoagulation: randomized study of the hemostatic effect on experimental hemorrhagic ulcer in dogs]. / La photocoagulation par laser Yag et l'électrohydrocoagulation monopolaire: étude randomisée de l'effet hémostatique sur l'ulcère hémorragique expérimental chez le chien.

The purpose of this randomized study was to compare the effects of two methods of hemostasis--photocoagulation using YAG Neodyme laser and liquid monopolar electrocoagulation--on acute experimental bleeding ulcers created in the dog stomach with an ulcer-maker. One hundred and fifty-three lesions were made and randomized into 3 groups; 51 lesions were treated with photocoagulation and complete hemostasis was achieved in all cases. Hemostasis was obtained in 80 p. 100 of 51 ulcers treated with liquid electrocoagulation. Control untreated ulcers remained hemorrhagic after 45 min of observation. The mean external muscle injury on day 7 was 55 p. 100 after photocoagulation and 65 p. 100 after liquid electrocoagulation. On day 14, mean external injury was 60 p. 100 after photocoagulation and 75 p. 100 after liquid electrocoagulation (non-significant difference). On day 7, the mean re-epithelization index, expressed as the percentage of the original ulcer diameter, ranged from 8 to 10 p. 100 in each trial group. On day 14, reepithelization covered 78 p. 100 of control ulcers and 72 p. 100 of photocoagulated ulcers (NS). This percentage falls to 47 p. 100 in ulcers treated with liquid electrocoagulation (p less than 0.01 when compared with ulcers treated with photocoagulation). Photocoagulation seemed to be more efficient in ensuring hemostasis and external muscle injury was correlated with the energy delivered. External muscle injury could not be controlled by liquid electrocoagulation. However the difference in the percentages of mean external muscle injury between the two methods was not significant. Therefore, in man, the risk of perforation is certainly slight and not very different whatever the method of hemostasis considered.

[Cytologic demonstration of immunoreactivity characteristic of adenopituitary peptides in the digestive epithelium of the rat and man]. / Mise en évidence cytologique d'immunoréactivités caractéristiques de peptides adénohypophysaires dans les épithéliums digestifs du rat et de l'homme.

Using immunoperoxidase techniques, the possible localization of pituitary regulatory peptides in fundic, antral and duodenal mucosae was investigated in both rat and man. All results obtained were similar in the two species. No glycopeptide (FSH, LH, TSH) was detected in the digestive tract. With different antisera directed against beta-lipotropin, alpha-MSH, beta-MSH, endorphins, ACTH 1-24, ACTH 17-39, a positive reaction was only obtained in the antral mucosae with an antiserum specific for the synthetic fragment 17-39 of ACTH. However neither the common precursor, proopiomelanocortin, nor the complete sequence of ACTH seem to be present in endocrine cells of the digestive tract. On the other hand, three antisera, directed against human growth hormone (GH), visualized numerous endocrine cells scattered in the glandular epithelium of the fundic and antral mucosae. Most cells were identified as ECL type in the gastric mucosae. Others are probably of the gastrin cell type in the antral mucosa, since these cells could be visualized on adjacent sections either with the antiserum against GH, or with a specific antiserum for gastrin.

Histological variations of the duodenal mucosa in chronic human pancreatitis.

From 16 volunteers with chronic pancreatitis and 36 healthy subjects duodenal biopsies were taken 15--20 cm beyond the papilla of Vater. Several morphometric parameters were calculated. The main results show: a significant decrease of villous area and height but not of the number of intestinal villi; a significant increase of Paneth cells; and a slight decrease in the number of glandular mitoses. This study suggests in man, a possible relationship between exocrine pancreatic secretion and the intestinal mucosa and a trophic action of pancreatic juice on the proliferation and the differentiation of the intestinal mucosa.

[Immunohistochemical localization of pituitary peptides in the rat pyloric antrum mucosa]. / Localisation immunohistochimique de peptides hypophysaires au niveau de la muqueuse antrale du rat.

Several peptides normally produced in the anterior pituitary lobe were searched in rat antral mucosa by immunocytochemistry. Peptides derived from pro-opiomelanocortin were tested: some endocrine cells were immunoreactive with ACTH 17-39 antiserum, and only a few elements were stained with ACTH 1-24 and beta LPH antisera. No immunoreactive cells were observed using alpha MSH, beta MSH and beta endorphine antisera. Using an antiserum against beta endorphin, a few cells and nerve fibres were immunostained. The other pituitary hormones were also tested: numerous antral cells contained immunoreactive GH, and some cells immunoreactive PRL or compounds chemically related to these hormones. No cells were stained with antisera directed against glycoproteic hormones. This work showed that several peptides previously localized in the pituitary gland were found in the antral mucosa. Further studies are needed to identify the cell types containing these peptides and to determine their origin.

Experimental study of laser photocoagulation in the treatment of bleeding canine gastric ulcers.

We report experimental results obtained by laser photocoagulation on hemorrhagic lesions induced in dogs. The aims of this study were 1) to evaluate the real efficiency of the YAG laser to ensure hemostasis of briskly bleeding lesions; 2) to determine the minimum energy density necessary to ensure hemostasis; 3) to assess the damage depth inside the gastric wall after photocoagulation; 4) to compare reepithelialization of treated ulcers with that of control ulcers; 5) to evaluate the safety margin for the clinical use of the YAG laser.

Long term effects of H2-receptor antagonists (cimetidine and ranitidine) on the human gastric and duodenal mucosa.

Cytological effects of two H2-receptor antagonists on the gastric and duodenal mucosa were studied during therapy of active duodenal ulcer (D.U.) in man. After endoscopic diagnosis (day 0), subjects were treated with cimetidine or ranitidine and re-examined on day 30. Only subjects with healed D.U. on day 30 were retained in this study. Gastric, pyloric and duodenal endoscopic biopsies were taken and treated for further morphometrical analysis both by light and electron microscopy. The use of the immunoperoxidase technique allowed evaluation of G and D cell populations. Kinetic parameters in proliferative zones were measured after in vitro incubation of biopsies with 3H-thymidine. No differences could be seen between the two H2-receptor antagonists. Increase of tubulovesicles and decrease of canaliculi in parietal cells are closely related to the inhibitory effect of these drugs on acid secretion. However secretory capacities of parietal cells are preserved since the whole membrane (tubulovesicle + canaliculi surface) remained constant. The collapsed aspect of the tubulovesicles on day 30 and the presence of connections between the tubulovesicle membrane, with both vesicle and canaliculi membrane, could support the theory of osmotic membrane expansion during parietal cell acid secretion. H2-receptor antagonists have been shown to be trophic in duodenal mucosa: both villi and microvilli area are increased. Confirming these findings, the proliferative compartment in the intestinal crypts was shown to be enlarged. No variation of G cell number could be seen in the antral mucosa; clear intracytoplasmic granules were increased in D.U. on day 0 but were not further modified on day 30. Somatostatin labelling index decreased. These last findings associated with the lack of G cell variations, suggest the presence of a possible paracrine modulation in the gastric and duodenal mucosa.