Prospective comparison of the value of brushings before and after biopsy in the endoscopic diagnosis of gastroesophageal malignancy

The hypothesis that the cytodiagnostic results on endoscopic brushings obtained before biopsy may be superior to those brushings obtained after biopsy, but with the accuracy of the subsequent biopsy reduced, was examined for 300 consecutive patients, including 256 with esophageal or gastric carcinomas. Following stratification by site and endoscopic appearance of their lesions, the patients were randomized to undergo brushing either before or after forceps biopsy. The accuracy of brushing cytology in patients with carcinoma was significantly higher when the brushing was performed before biopsy than after biopsy (93.5% versus 82.6%; P less than .01). The diagnostic yield of the biopsy was not significantly different whether the lesions were brushed before or after the biopsy (92.7% versus 93.2%; P less than .5). The diagnostic superiority of brushings obtained before biopsy did not relate to the site or endoscopic appearance of the tumor. There were no false-positive cytologic or histologic reports. For all 256 carcinomas, the cumulative accuracy (brushing cytology plus biopsy) reached 98.8% and was significantly better (P less than .001) than that of biopsy alone (93.9%) or cytology alone (87.9%). Apart from reinforcing the belief that the combined application of brushing and biopsy is mandatory for achieving optimal results, this study indicated that the brushing should be performed before the biopsy.     

Endoscopic fine needle aspiration biopsy

A catheter assembly that can be passed through the biopsy channel of a standard fiberoptic gastroduodenoscope was devised to permit fine needle aspiration biopsy of gastrointestinal neoplasms under direct vision of the endoscope. This technique for endoscopic aspiration biopsy was performed in ten consecutive patients with esophageal and gastric carcinomas, along with the conventional endoscopic brushing and biopsy. Endoscopic aspiration biopsy gave a positive diagnosis in all ten cases while the other two techniques gave inconclusive results in one patient with an ulcerative growth. We feel that endoscopic aspiration biopsy can be used to obtain representative samples from gastrointestinal neoplasms, and it may add to the diagnostic accuracy of endoscopic biopsy and brushing cytology.     

The production and characterization of monoclonal antibodies to a human colonic antigen associated with ulcerative colitis: cellular localization of the antigen by using the monoclonal antibody

We detected in human colon extracts a 40 kDa protein(s) that specifically reacts with tissue-bound IgG obtained from the colon of patients with ulcerative colitis or CCA-IgG. Using the hybridoma technology, we developed monoclonal antibodies to this 40 kDa protein. The specific immunoreactivity of one of the monoclonal antibodies (7E12H12, IgM isotype) against the 40 kDa protein was demonstrated both by ELISA and by immunotransblot. Competitive binding experiments showed that CCA-IgG inhibits the binding of 7E12H12 to the 40 kDa protein, suggesting the recognition of common epitope(s) on the 40 kDa protein by the monoclonal antibody and CCA-IgG. 7E12H12 was used to determine cellular localization of the 40 kDa protein. Biopsy tissue specimens from colon, esophagus, stomach, duodenum, jejunum, ileum, liver, pancreas, lungs, kidneys, salivary, and mammary glands were obtained. Tissue specimens were fixed in 4% paraformaldehyde or in 10% formalin. Sections were sequentially incubated with the hybridoma supernatant, biotinylated anti-mouse IgM, avidin-biotin-peroxidase complex, and 3,3'-diaminobenzidine. An unrelated hybridoma supernatant was used as control. The monoclonal antibody exclusively recognized colonic epithelial cells both in the crypt and on the luminal surface. Immunoreactivity was present on the plasma membrane chiefly along the basolateral areas of the cells. Plasma membrane localization of the 40 kDa protein was confirmed by immunoelectron microscopy. All colonic mucosal biopsy specimens from both adult and fetal colon reacted with the monoclonal antibody. None of the biopsy specimens from stomach, duodenum, jejunum, ileum, liver, pancreas, or non-gastrointestinal tissue reacted with the antibody, confirming the organ specificity of the 40 kDa protein. The interaction between this colonic epithelial membrane protein and the CCA-IgG may play an important role in the pathogenesis of ulcerative colitis.     

Rumenectomy-induced proliferation in duodenal villous epithelium is mechanistically related to the disappearance of statin, a non-proliferation-specific nuclear protein

We undertook this study to determine the effect of rumenectomy (a known cause of duodenal crypt cell hyperplasia) on the epithelial growth kinetics of the crypt-villus axis in rat duodenum. Ten rats were randomly assigned to control (gastrotomy) and experimental (rumenectomy) groups. After 14 days rats were sacrificed and representative sections were stained with the monoclonal antibody to statin, a non-proliferation-specific protein, by the immunoperoxidase procedure. In the control group, the mean percentages of statin-positive cells in the proximal duodenum, distal duodenum, proximal jejunum, and distal jejunum were 79 +/- 8.5, 79.5 +/- 5.7, 85 +/- 1.4, and 83.5 +/- 0.7, respectively. In the rumenectomy group, statin-positive nuclei were found in the region of the villous apices only, and the corresponding values for the above four areas were 26.2 +/- 4.9, 24.5 +/- 3.5, 31.7 +/- 4.5, and 80.5 +/- 2.1. Except for distal jejunum, the differences in statin expression in the control and experimental groups were significant (p less than 0.001). Rumenectomy leads to the disappearance of statin from the villous column cells of the duodenum and proximal small bowel. The lack of expression of statin in the rumenectomy group documents the potential usefulness of this measure in future studies in neoplasia were understanding of the proliferative status is of crucial importance.     

An Enzyme Histochemical Investigation of the Intestinal Mucosa in Diarrheic Calves

Selected enzymes were examined in the small intestine of twelve 2–5 week-old calves, 8 with diarrhea and 4 convalescents. The diarrheic calves showed a reduction of enzyme reactions mainly in the duodenum and middle small intestine, and the crypt reactions appeared most severely affected. In the duodenum, villous alkaline phosphatase, adenosine triphosphate-(ATP)-splitting enzyme, and β-D-galactosidase were reduced in 3 calves; the reaction in the corresponding crypts was decreased in 6 calves for the ATP-splitting enzyme and in 4 calves for the β-D-galactosidase. Six calves showed decrease of villous brush border acid phosphatase, and 3 of villous non-specific esterase. In the middle jejunum, villous ATP-splitting enzyme was reduced in 3 calves, while 5 showed decrease of the corresponding crypt reaction. Convalescents had no enzyme reduction in the duodenum, whereas 1 showed marked reduction of the ATP-splitting enzyme and aminopeptidase in the middle and posterior jejunum. The decreased enzyme reactions in the present material may be caused by immaturity of epithelial cells associated with regenerative crypt hyperplasia and/or microbial destruction of enzymes.     

Gluten-sensitive enteropathy in a cynomolgus monkey

A malabsorption syndrome was observed in a cynomolgus macaque. Clinical signs included weight loss despite increased appetite, and diarrhea, characterized by an increased volume of soft, tan, malodorous feces. Clinicopathologic findings included hypoalbuminemia, generalized dilation of bowel loops with a prolonged transit time, steatorrhea and markedly diminished absorption of D-xylose. Biopsies of the duodenum and jejunum had total villous atrophy, crypt hyperplasia and a plasmacytic-lymphocytic infiltrate of the lamina propria. The monkey's diet was changed to a semi-synthetic diet containing no grain products. Subsequently, stool characteristics, body weight and intestinal villous morphology returned to normal. This response to removal of grain products from the diet suggests a syndrome similar to gluten-sensitivity enteropathy in human beings.     

Stereological studies on the small intestinal epithelium of the rat

Summary Quantitative macroscopic, light-microscopic and electron-microscopic studies were performed on the small intestine of fasted and non-fasted adult, male Sprague-Dawley rats. In non-fasted rats the small intestine was longer than in fasted rats. Due to the presence of villi the surface area in the duodenum and the jejunum was enlarged about six times. The microvilli on the villous crests caused a surface enlargement by 13 times in the duodenum (value corrected for overestimation due to section thickness), and 19 times in the jejunum of the fasted rats. At the base of the villi these values were about 50% lower. It was calculated that, in the fasted rats, the total enlargement of the luminal surface area — due to villi and microvilli — was 63 times in the duodenum and 81 times in the jejunum (corrected for section thickness).Differences between the villous crest epithelium and the villous base epithelium were also found with regard to the mean cell height, and the volume densities of the absorptive cell nuclei, the mitochondria, and the paracellular channels.Supported by grants from the Swedish Medical Research Council (Project No. 12X-2298), from the Swedish Group-Insurance Co. Förenade Liv, from Tore Nilson's Fund for Medical Research and from the Medical Faculty, University of Umeå     

Detection of continuing gluten ingestion in treated coeliac patients.

To assess the incidence and effects of continuing gluten ingestion in coeliac disease 51 adult coeliac patients were studied after four to 132 (mean 63) months on a prescribed gluten-free diet. Each patient completed a prospective dietary questionnaire, underwent a repeat jejunal biopsy, and gave serum for gluten antibody estimation. Altogether 65% of patients were still ingesting gluten, often inadvertently. Direct questioning on dietary habits had failed to uncover most of this consumption. The gluten antibody test proved a useful screening test for detecting continuing gluten ingestion and patients with both persistent subtotal villous atrophy and gluten antibodies were almost certain to be taking large amounts ( more than 2 g/day). The presence of persistent partial villous atrophy was found, however, to be an unreliable guide to gluten intake.     

Endoscopically directed fine needle aspiration biopsy of gastric and esophageal lesions.

Endoscopically directed fine needle aspiration (FNA) has been reported to be a valuable adjunct to forceps biopsy in the evaluation of gastric and esophageal lesions. In our series of 38 cases with endoscopically detected mucosal and submucosal abnormalities, FNAs were obtained with a Stifcor transbronchial aspiration needle. Four cases were reported as insufficient. Five aspirates correctly documented the presence of a neoplasm, but three failed to identify a subsequently histologically confirmed adenocarcinoma. Two cases were falsely suspicious for adenocarcinoma. FNA correctly excluded lymphoma in 12 patients with thick gastric folds clinically suspicious for lymphoma. FNA is a useful adjunct to forceps biopsy of neoplastic and inflammatory lesions in both mucosal and submucosal locations within the upper gastrointestinal tract.     

Bronchoscopy in the European hamster (Cricetus cricetus).

A pediatric bronchoscope was modified for use in the European hamster (Cricetus cricetus) and has been used successfully to obtain biopsy specimens from these animals. Biopsy specimens were obtained by means of small forceps with visual control given by the modified bronchoscope.     

Effect of Weaning on Small Intestinal Structure and Function in the Piglet

Fifty-four piglets were selected from 12 litters weaned at 17 (Treatment 1), 21 (Treatment 2), 28 (Treatment 3) and 35 (Treatment 4) days old, respectively, to determine the effect of weaning age on small intestinal villus morphology, immunology and histochemistry. From proximal duodenum, proximal jejunum, distal jejunum and middle ileum, intestinal samples with three replicates (piglets) in each treatment were taken at 18, 22, 28 and 36; 22, 28, 36 and 43; 28, 36, 43, and 50; and 18, 22, 28, 36, 43 and 50d of age in Treatment 1, 2, 3 and 4, respectively. This was equivalent to 12h, 3d, 1 week, 2 week postweaning in Treatment 1; 12h, 1 week, 2 week, 3 week postweaning in Treatment 2 and 3, and all the same age in Treatment 4 as in Treatment 1, 2, 3, respectively. The results showed that villous height of duodenum and proximal jejunum decreased significantly in Treatment 1 and 3. Crypt depth in the duodenum, proximal jejunum and ileum also decreased significantly in Treatment 1. Date had significant effect on villous height of the duodenum, distal jejunum and ileum with the shortest on day 29 and crypt depth of all positions increased with piglet age except the crypt depth in proximal jejunum decreased on day 50. Weaning age and day of age had significant effects on intraepithelial lymphocyte (IEL) number and goblet cell (GC) number at all positions of small intestinal mucosa in piglets. The number of IEL at all segments of small intestinal mucosa in Treatment 3 increased significantly compared to those in other treatments, but IEL number at all locations of small intestinal mucosa in Treatment 2 decreased significantly compared to those in other treatments. The number of GC in small intestinal mucosa increased significantly in early-weaned (< day 21) piglets. It appears that providing fluid milk replacer for a few days postweaning could dramatically reduce the negative impact of weaning on villous morphology and digestive and absorptive function, especially in pigs weaned prior to 3 week of age. Finally, as weaning age was reduced, GC had a greater role in intestinal duct protection.     

Appearance of interstitial cells of Cajal in the human midgut

Several subtypes of the interstitial cells of Cajal (ICC) form networks that play a role in gastrointestinal motor control. ICC express c-kit and depend on signaling via Kit receptors for development and phenotype maintenance. At 7-8 weeks of development, c-kit-immunoreactive (c-kit-IR) cells are present in the human oesophagus, stomach and proximal duodenum wall. In the remaining small and large bowel, c-kit-IR cells appear later. The object of the present study is to determine the timing of the appearance of c-kit-IR ICC in the parts of the digestive tube originating from the midgut (distal duodenum, jejunum, ileum and proximal colon). Specimens were obtained from eight human embryos and 11 fetuses at 7-12 weeks of gestational age. The specimens were exposed to anti-c-kit antibodies to investigate ICC differentiation. The differentiation of enteric neurons and smooth muscle cells was immunohistochemically examined by using anti-PGP9,5 and anti-desmin antibodies, respectively. In the distal duodenum, jejunum and ileum, c-kit-IR cells emerged at week 9 at the level of the myenteric plexus in the form of a thin row of cells encircling the inception of the ganglia. These cells were multipolar or spindle-shaped with two long processes and corresponded to the ICC of the myenteric plexus. In the proximal colon, c-kit-IR cells emerged at week 9-10 in the form of two parallel belts of cells extending at the submucosal plexus and the myenteric plexus levels. We conclude that ICC develop following two different patterns in the human midgut.     

Culture of Helicobacter pylori: Effect of Preimmersion of Biopsy Forceps in Formalin

Background. Treatment of antibiotic-resistant Helicobacter pylori should be based on bacterial sensitivity testing that requires the ability to isolate the bacterium from gastric mucosal biopsies. The aim of this study was to determine whether the yield for detecting H. pylori infection by culture is reduced by immersion of biopsy forceps in formalin prior to obtaining the specimen.
Materials and Methods. Gastric antral mucosal biopsies (100 specimens) from 50 patients were obtained for culture of H. pylori. An antral biopsy was taken for culture, and with the same forceps a biopsy was taken for histological examination. The biopsy specimen was removed by shaking, whereas the forceps was immersed in 10% buffered formalin for the histological investigation. The forceps was then used without rinsing to obtain a second specimen for culture from an area adjacent to the first site. H. pylori status was determined by histological assessment with the Genta stain and a rapid urease test.
Results. Fifty patients with H. pylori infection documented by histological inquiry and positive rapid urease testing entered the study; 29 had duodenal ulcers, 5 had gastric ulcers, 1 had mucosal associated lymphoid tissue (MALT) lymphoma, and 15 were without ulcer disease. The results of culture both before and after immersion in formalin were identical. One patient had both cultures negative; the sensitivity of culture for detection of H. pylori infection was 98% (95% confidence interval =93%-100%).
Conclusion. Preimmersion of biopsy forceps in formalin does not adversely affect the ability to culture H. pylori.     

Diagnosis of abdominal masses with percutaneous biopsy guided by ultrasound.

OBJECTIVE--To assess the accuracy and safety of percutaneous biopsy of abdominal masses guided by ultrasound. DESIGN--Prospective study. SETTING--Combined gastroenterology service, Scarborough Hospital. PATIENTS--108 Consecutive patients identified as having a discrete mass on diagnostic ultrasound examination of the abdomen. INTERVENTION--A sample of tissue was obtained with an aseptic technique under local anaesthesia: an 18 steel wire gauge needle (Tru-Cut) was mounted in a spring loaded firing device (Biopty gun) that was advanced under simultaneous ultrasound scanning, permitting precise localisation of the target organ. MAIN OUTCOME MEASURE--Results of histological examination of tissue specimens. RESULTS--Biopsy failed in four patients. Adequate histological specimens were obtained in 104 patients with masses in the liver (31), pancreas (37), kidney (10), and adrenal glands (six) and in 20 undiagnosed abdominal and retroperitoneal masses. Follow up was until death or confirmation of the diagnosis. Three complications but no deaths occurred. Malignancy was suspected in 84 patients before biopsy. This was confirmed in 70 patients, in 26 of whom confirmation of dissemination obviated the need for further investigation. In 10 patients biopsy indicated a previously unsuspected primary tumour, and in 12 it showed only a benign lesion. Among 24 patients considered to have benign disease biopsy showed an unsuspected neoplasm in seven. Use of biopsy thus had a major effect on clinical management in 55 patients. Four false negative but no false positive diagnoses resulted from the procedure. CONCLUSION--Percutaneous biopsy of abdominal and retroperitoneal masses under ultrasound guidance is a safe and accurate method of obtaining a histological diagnosis. The results obtained have a considerable effect on clinical management.     

Dietary alanyl-glutamine improves growth performance of weaned piglets through maintaining intestinal morphology and digestion–absorption function

Alanyl-glutamine (Ala-Gln), a highly soluble and stable glutamine dipeptide, is known to improve gut integrity and function. The aim of this study was to evaluate whether dietary Ala-Gln supplementation could improve growth performance, intestinal development and digestive-absorption function in weaned piglets. A total of 100 purebred Yorkshire piglets weaned at 21 days of age were assigned randomly to four dietary treatment groups and fed a basal diet (control group) or a basal diet containing 0.15%, 0.30% and 0.45% Ala-Gln, respectively. Compared with the control group, piglets fed the Ala-Gln diets had higher average daily gain and lower feed : gain and diarrhea rate (P < 0.05). Moreover, dietary Ala-Gln supplementation increased villous height and villous height : crypt depth ratio in duodenum and jejunum (P < 0.05), as well as the activities of maltase and lysozyme in jejunum mucosa (P < 0.05). In addition, a decrease in serum diamine oxidase activity and crypt depth in duodenum and jejunum was observed in piglets fed the Ala-Gln diets (P < 0.05). Serum cytosolic phospholipase A2 (cPLA2) concentration and gene expression of cPLA2, Na⁺-dependent glucose transporter 1, glucose transporter 2 and peptide transporter 1 in jejunum were increased by feeding Ala-Gln diets relative to control diet (P < 0.05). These results indicated that feeding Ala-Gln diet has beneficial effects on the growth performance of weaned piglets, which associated with maintaining intestinal morphology and digestive-absorption function.     

Endoscopic brushing cytology and biopsy in the diagnosis of upper gastrointestinal tract lesions. A study of 350 cases

Direct-vision endoscopic examination conducted on 4,000 patients for persistent upper gastrointestinal (GI) complaints over a period of five years revealed 350 visible lesions that were subjected to brushing cytology and biopsy. Cytologic examination of brushing smears from all 350 cases showed malignant cells in 67 (19.14%), cells suggesting benign polypoid neoplasms in 4 (1.14%), ulcerative and reparative features with attendant atypias in 186 (53.14%), inflammatory findings in 91 (26%) and false-negative findings in 2 cases (0.57%). Only 259 (74%) of the visible lesions were also subjected to endoscopic biopsy. Of the 67 patients with positive cytology, 52 were judged positive on the biopsy specimen; the 2 false-negative cytologic reports were confirmed as positive by biopsy. In four patients with gastric ulcers, malignant cells were seen along with gastric repair cells. This study indicates that brushing cytology is very useful in detecting benign ulcerative lesions with their atypias, a feature that could be useful in monitoring and controlling lesions in high-risk groups of patients, such as in India. In this study, endoscopic brushing cytology gave a better diagnostic yield than did endoscopic tissue biopsy. However, the two techniques are complementary for the diagnosis of upper GI malignancies.     

Survival of cultured allografts in patients with burns assessed with probe specific for Y chromosome.

The aim of the study was to determine the fate of cultured skin allografts in patients with burns. In situ DNA hybridisation with a Y probe (pHY 2.1) was used to detect cells carrying the Y chromosome (the probe being visualised by the alkaline phosphatase-antialkaline phosphatase method) in biopsy specimens taken from cultured allografts derived from donors of the opposite sex to the recipients (20 patients with burns). Specimens were taken within a week, between one and three weeks, between four and six weeks, and more than six weeks after grafting. Only two of the 27 biopsy specimens contained cells that were the same sex as the donor; both were taken within a week after grafting. In the 25 other specimens the epithelial cells were the same sex as the recipient. Cultured skin allografts showed no evidence of survival in patients with burns, which suggests that they are probably not suitable for long term management of burns but may be useful as short term biological dressings.     

Duodenoscopy in the diagnosis of upper gastrointestinal disease

Duodenoscopy was undertaken in 143 patients with upper gastrointestinal tract symptoms. There were no complications of the procedure. The duodenum was successfully entered in 98% of attempts, and in 90% of patients the examination was judged to be technically successful. When compared with radiological assessment, the findings at duodenoscopy significantly altered diagnosis and management in 21% of patients. The endoscopic appearance of the non-ulcerated duodenal mucosa could not be consistently correlated with changes found in biopsy tissue obtained under direct vision. Duodenoscopy is a valuable adjunct in the clinical assessment of upper gastrointestinal tract symptoms suspected to be due to pathological changes in the duodenum.     

Arginine-vasopressin immunoreactive material in the gastrointestinal tract

Like many other neuropeptides, vasopressin is not confined to the hypothalamic neurohypophysial system. Furthermore, vasopressin was found to be a potent vasoconstrictor in the rat jejunum, reducing myenteric artery flow. These associations were the basis of this investigation on the presence of vasopressin in the gastrointestinal (GI) tract by both RIA and immunohistochemistry. Portions of the gastrointestinal tract and pancreatic islets of the rat were extracted with 0.1 N HCl for RIA measurements of AVP content. Similar portions from the male cat GI tract were used for immunohistochemistry studies. Acid extracts of the GI tract were found to contain immunoreactive AVP with the highest concentration (pg/mg protein) in the fundus portion of the stomach (15.0 +/- 1.6) and slightly lower values down along the antrum-pylorus portion (6.7 +/- 0.6), proximal jejunum (8.6 +/- 0.2), distal ileum (9.7 +/- 0.3) and colon (11.9 +/- 0.5). In the pancreatic islets the concentration was much higher (72.0 pg/mg protein). The extract inhibition curves showed parallelism with the appropriate standard preparation of AVP in the specific RIA. Immunohistochemical localization showed IR-AVP in the nerve fibers around the myenteric plexus of the second portion of the duodenum. It was also found in fibers starting from where the myenteric plexus goes through the layer of muscle fibers, penetrating the submucosa and duodenal mucosa, ending near the capillaries situated along the basal side of the villous epithelium cells. Similar IR-AVP activity was found in cells located in the mucosal epithelium of the duodenum, jejunum, ileum, colon and rectum.(ABSTRACT TRUNCATED AT 250 WORDS)