Histochemical demonstration of gastrointestinal mucins in ovarian mucinous cystadenoma.

Forty-one specimens of mucinous cystadenoma (MCA) of the ovary and 13 specimens of normal uterine endocervix were investigated histochemically using alcian blue pH 2.5-PAS, high iron diamine-alcian blue pH 2.5, galactose oxidase-Schiff reaction (GOS, for detection of gastric surface mucous cell mucins), paradoxical Concanavalin A staining (PCS, for detection of gastric gland cell mucins) and periodic acid-sodium borohydride-potassium hydroxide-PAS (PA-SB-PH-PAS) reaction (for detection of mucins of the large intestine). In addition, Grimelius staining was carried out to explore the distribution of endocrine cells. The MCAs contained abundant neutral mucins but a little acid mucins, whereas sulfomucins predominated in normal endocervices. Among the MCAs, 30, 26, 2 and 9 had tumor cells positive for GOS, PCS, PA-SB-PH-PAS and Grimelius stain, respectively. These results indicate that MCAs frequently contain tumor cells of the gastrointestinal type and endocrine cell type. Endocervical epithelia, on the other hand, lack reactivity for PCS, PA-SB-PH-PAS and Grimelius stain, although in 8 of the present specimens, the lining cells showed weak GOS reactivity.     

Histochemical Demonstration of Gastrointestinal Mucins in Ovarian Mucinous Cystadenoma

Forty-one specimens of mucinous cystadenoma (MCA) of the ovary and 13 specimens of normal uterine endocervix were investigated histochemically using alcian blue pH 2.5 PAS, high iron diamine-alcian blue pH 2.5, galactose oxidase Schiff reaction (GOS, for detection of gastric surface mucous cell mucins), paradoxical Concanavalin A staining (PCS, for detection of gastric gland cell mucins) and periodic acid-sodium borohydride-potassium hydroxide PAS (PA SB-PH-PAS) reaction (for detection of mucins of the large intestine). In addition, Grimelius staining was carried out to explore the distribution of endocrine cells. The MCAs contained abundant neutral mucins but a little acid mucins, whereas sulfomucins predominated in normal endocervices. Among the MCAs, 30, 26, 2 and 9 had tumor cells positive for GOS, PCS, PA-SB-PH PAS and Grimelius stain, respectively. These results indicate that MCAs frequently contain tumor cells of the gastrointestinal type and endocrine cell type. En-docervical epithelia, on the other hand, lack reactivity for PCS, PA SB PH PAS and Grimelius stain, although in 8 of the present specimens, the lining cells showed weak GOS reactivity.     

Histochemistry of mucin secreting components in mucoepidermoid and adenosquamous carcinoma of the oesophagus.

AIMS--To determine the direction of differentiation of the mucin secreting components in a rare group of oesophageal tumours--oesophageal squamous cell carcinomas with prominent mucin secreting components (mucoepidermoid carcinomas and adenosquamous carcinomas). METHODS--In a review of 617 cases of primary carcinoma of the oesophagus, 16 cases of squamous cell carcinoma with prominent mucin secreting components were studied using a battery of histochemical techniques. RESULTS--The mucin produced by these tumours was mixed and included a variable content of enzyme labile sialomucin (positive for mucicarmine, periodic acid Schiff, and alcian blue, and sensitive to sialidase digestion and negative for high iron diamine-alcian blue). Retrospective analysis of endoscopic biopsy specimens taken from these tumours showed that mucin was present in five (42%) cases. CONCLUSIONS--The glandular component of this group of tumours histochemically differentiated in the direction of oesophageal glands: examination of the mucin secreting component in squamous cell carcinoma in resected specimens is therefore required for recording the true incidence of this type of tumour.     

Acidic mucin in the prostate: Can it differentiate adenosis from adenocarcinoma?

Numerous reports have claimed that because acidic mucin is absent in benign prostatic glands and is present in some prostatic adenocarcinomas, this stain may be an adjunctive aid in the diagnosis of adenocarcinoma of the prostate. However, adenosis that mimics low-grade adenocarcinoma has not been evaluated to date. We studied 28 foci of adenosis for the presence of high iron diamine-alcian blue (HID-AB). Fifteen foci of adenosis (54%) showed strong staining for HID-AB; staining was diffuse in 11 cases and focal in four cases. An additional two cases (7%) showed equivocal staining. The remaining 11 cases (39%) lacked positivity. All cases of adenosis were verified with immunohistochemistry for keratin 903, a basal cell-specific antibody. This study demonstrates the limited use of acid mucin staining in the diagnosis of adenocarcinoma. The finding of HID-AB positivity in occasional isolated benign small prostatic glands within hyperplastic nodules suggests that acid mucin secretion may be a reflection of gland size or proliferation rather than evidence that adenosis is related to adenocarcinoma of the prostate.     

Predominance of sialomucin secretion in malignant and premalignant pancreatic lesions.

Sialomucin and sulphomucin-secreting cells were studied in the normal and pathologic human pancreas with the high iron diamine-alcian blue technique which allows differentiation between the two types of mucin. In six normal autopsy pancreata, only sulphomucin was found. In benign lesions of either calcifying chronic pancreatitis (seven cases) or obstructed chronic pancreatitis (six cases), sulphomucins were widely predominant. In contrast, malignant lesions (pancreatic adenocarcinoma [12 cases], cystadenocarcinoma [two cases]) or premalignant lesions (mucinous cystadenoma [one case], ductectatic mucinous cystadenoma [four cases], villous adenoma of the main pancreatic duct [two cases]) showed a predominant sialomucin secretion, except for three poorly differentiated pancreatic carcinomas that did not show mucin staining. The sialomucin positivity was not observed at distance from the malignant lesions. In one case of benign enteroid cyst, sulphomucins predominated. These findings indicate a preponderance of sialomucin secretion in malignant or premalignant pancreatic lesions.     

Hematoxylin and eosin staining of mucins of the gastrointestinal tract

An infrequent observation of assessing hematoxylin and eosin sections is the blue staining of mucins (for example those in goblet cells). This is believed to be due to a low concentration of alum and high pH of the hematoxylin staining solution. This study examines the incidence of blue mucin in various sites of the gastrointestinal tract using a low alum, high pH hematoxylin solution. The results are compared with a conventional hematoxylin solution, iron alum celestine blue method and an alcian blue (pH 2.5)-periodic acid-Schiff (AB-PAS) stain to characterize the type of mucin demonstrated. This study is the first to offer evidence that blue-stained mucin with low alum, high pH hematoxylin corresponds with carboxylated mucins as shown by the AB-PAS stain in the gastrointestinal tract. Iron alum celestine blue was also found to stain the mucin of a proportion of rectal biopsies and appendix as well as the carboxylated mucin of one duodenal biopsy.     

Sulphomucins favour adhesion of Helicobacter pylori to metaplastic gastric mucosa.

AIMS: To assess the influence of sulphomucin secretion on Helicobacter pylori colonisation and adhesion to metaplastic gastric cells. METHODS: Gastric biopsies from 230 H pylori positive patients with intestinal metaplasia were analysed. Sulphated mucins and H pylori were visualised using a new technique combining high iron diamine-alcian blue mucin stains with the Steiner silver stain for the bacteria. RESULTS: Sulphomucin secretion anywhere in the mucosa and a histological diagnosis of dysplasia increase the risk of H pylori adhesion to metaplastic cells (odds ratios 19.9 and 4.3, respectively). However, only 9.4% of cases showing sulphomucin secretion and 10.8% of cases with dysplasia had evidence of adhesion of H pylori bacteria to metaplastic cells. CONCLUSIONS: The findings suggest that H pylori may play a role in the advanced stages of carcinogenesis. It will be of interest to investigate if the relative small proportion of type III metaplasias that actually progress to carcinoma show persistence of H pylori.     

Comparison between pH 2·5 Alcian blue stain and Hale’s dialyzed iron technique for demonstrating acid mucin in colorectal carcinoma

Abstract

This study compared the value of pH 2·5 Alcian blue and Hale’s dialyzed iron methods for demonstrating acid mucin in colorectal carcinoma. A total of 40 colorectal biopsies from known positive cases were examined. Sections of the biopsies were stained using pH 2·5 Alcian blue method and Hale’s dialyzed iron technique, and classified as ‘good’ or ‘excellent’ according to staining intensity and efficiency, using a semi-quantitative scoring system. The pH 2·5 Alcian blue method was found to provide a higher frequency of ‘excellent’ scores compared to Hale’s dialyzed iron technique, 32 (80%) versus 21 (52.5%) respectively (P = 0·018). Both methods provide at least a minimal level of satisfactory staining results and seem to demonstrate the same amounts of acid mucins; however, if high quality Alcian blue dyes are available, pH 2·5 Alcian blue method is especially recommended to stain colorectal carcinoma acid mucins because it provides a stronger staining intensity and coloration compared to Hale’s dialyzed iron technique.     

Alterations in mucin type: an indicator for suspicion of malignant gastric transformation

Mucins are produced by both benign and malignant gastric epithelium. In general, mucins can be classified into neutral and acidic mucins. The latter are of 2 major types, sulphated (sulphomucins) and carboxylated (sialomucins). A retrospective study was initiated at the Department of Pathology, University Hospital, Kuala Lumpur to histochemically study the mucin profiles of cases of intestinal (IGC) and diffuse (DGC) types of gastric carcinoma in Malaysian patients to determine whether a significant change of mucin type occurs in the event of malignant transformation. 42 IGC and 37 DGC were subjected to alcian blue-periodic acid Schiff and high iron diamine-alcian blue histochemical staining. In addition, 18 cases of gastrectomies performed for benign lesions in the stomach served as normal controls. The number of cases of IGC and DGC which exhibited sulphomucin production was significantly increased (p < 0.001) compared to normal controls. Also, the number of cases of DGC which produced neutral mucin were significantly less (p < 0.05) than the control group. However, there was no significant difference between the number of IGC and DGC cases which demonstrated sialomucin production and normal controls. It appears that while not pathognomonic, a lack of neutral mucin production should alert the pathologist to the possibility of a gastric malignancy, in particular DGC. The likelihood of a malignant lesion would be further supported if there is an increased sulphomucin production.     

Gastric carcinoma metastatic to the breast diagnosed by mucosubstance histochemistry and electron microscopy

Bilateral breast tumors with a histologically scirrhous pattern were conclusively diagnosed as gastric carcinoma metastatic to the breast using mucosubstance histochemistry and electron microscopy. The majority of the carcinoma cells gave histochemically positive reactions for galactose oxidase-Schiff, stable class III con A, and high-iron diamine-alcian blue (HID-AB). Also electron microscopically, numerous HID-positive mucus droplets (sulfomucin) were seen in the carcinoma cells. These findings invalidated the possibility that the carcinomas were primary breast cancer. The practical applications of distinctive patterns of mucus secretion are discussed.     

GASTRIC CARCINOMA METASTATIC TO THE BREAST DIAGNOSED BY MUCOSUBSTANCE HISTOCHEMISTRY and ELECTRON MICROSCOPY

Bilateral breast tumors with a histologically scirrhous pattern were conclusively diagnosed as gastric carcinoma metastatic to the breast using mucosubstance histochemistry and electron microscopy. The majority of the carcinoma cells gave histochemically positive reactions for galactose oxidase-Schiff, stable class III con A, and high-iron diamine-alcian blue (HID-AB). Also electron microscopically, numerous HID-positive mucus droplets (sulfo-mucin) were seen in the carcinoma cells. These findings invalidated the possibility that the carcinomas were primary breast cancer. The practical applications of distinctive patterns of mucus secretion are discussed. ACTA PATHOL JPN 38: 1353-1361, 1988.     

Carbohydrate cytochemistry of rhesus monkey tracheal submucosal glands

This study was designed to characterize the ultrastructure and carbohydrate content of secretory cells in submucosal glands of rhesus monkey and to compare this information with that available for humans. The tracheas from five adult monkeys were fixed by airway infusion, processed, and embedded for both light and transmission electron microscopy. Histochemical strains including alcian blue-periodic acid-Schiff, dialyzed iron, and high-iron diamine-alcian blue were applied to serial glycol methacrylate sections. The cytochemical stains used included periodic acid-thiocarbohydrazide-silver proteinate, high-iron diamine, and low-iron diamine. The glandular secretory cells were divided into four categories based on ultrastructure and location within the gland. Cells in the first category resembled the mucous cell of the surface epithelium and were located in ducts most proximal to the tracheal lumen. The second category consisted of cells that were located in distal ducts and contained large electron-lucent granules. The granules in both of these cell groups contained material that was periodate-reactive and sulfated. Cells of the third category contained granules that were either electron-lucent or electron-dense. These cells, which were difficult to characterize as either serous or mucous, were located in secretory tubules and acini and contained periodate-reactive glycoconjugates that were either sulfated or nonsulfated. The last category consisted mainly of cells that contained electron-dense granules that were lightly periodate-reactive or a few that were unreactive with any of the cytochemical methods used here.(ABSTRACT TRUNCATED AT 250 WORDS)     

Iron pyronine and alcian blue for staining acid mucin in plastic-embedded sections

Iron pyronine Y and Alcian blue demonstrated sulfated and nonsulfated acid mucin, respectively, in plastic-embedded sections. Safran used in combination with these two dyes stained collagen and some reticulum fibers. Sulfated acid mucin was red, while nonsulfated acid mucin stained blue; collagen appeared yellow to greenish yellow. The iron pyronine Y-Alcian blue-safran staining method, when used as in the present article, produces excellent cellular visualization of sulfated and nonsulfated acid mucin.     

Iron medication-induced gastric mucosal injury

Severe gastrointestinal erosion, ulcer, necrosis and strictures after an acute iron overdose are well described. However, gastric mucosal injury in patients receiving therapeutic iron has received only scant recognition despite its wide use. We report a case of iron medication-induced gastric mucosal injury in a 76-year-old male who presented with iron deficiency anemia and had been taking ferrous sulfate tablet for 4 years. Esophagogastroduodenoscopy (EGD) revealed a pale, villous appearing flat lesion along the lesser curvature of gastric body. Histopathologic examination of EGD biopsies of the flat lesion showed brown crystalline materials deposited in the lamina propria of gastric mucosa, which was accompanied with fibrosis, chronic inflammation, and foreign body reaction. The crystalline materials were covered and admixed with gastric epithelium. Prussian blue iron stain confirmed that the brown crystalline materials were iron. The iron and hemosiderin accumulation was also seen in cytoplasm of epithelial cells and lumen of fundic gastric glands. The recognition and reporting by pathologists of iron-induced changes in EGD biopsies will alert clinicians to this underrecognized but easily correctable complication by alternative forms of iron therapy, such as liquid preparation.     

Pathways of clearance in mouse lungs exposed to iron oxide aerosols

The history of particle clearance was studied in lungs of mice serially sacrificed at intervals up to 14 months following single exposures to an aerosol of submicronic, particulate, iron oxide used as a similitude for atmospheric dust. Clearance was followed by light microscopy in unstained and Prussian blue stained frozen and plastic embedded sections, as well as by electron microscopy, where iron oxide can be recognized by its form. Related problems were investigated through histochemical demonstration of acid phosphatase activity in pulmonary lysosomes and Prussian blue staining of various tissues after administration of iron compounds by gastrointestinal and vascular routes. The iron particles settle extensively but not uniformly on pulmonary alveolar surfaces. Clearance is centripetal and involves two mechanisms, an extracellular mechanism fed by fluid currents sweeping across the surface, and a cellular mechanism principally involving alveolar macrophages. In the early post exposure period both actively remove deposited particles predominantly through the pulmonary airways. By 24 hours uncleared residues have become ingested and clearance thereafter results mainly from cellular action. Macrophages enter bronchial passages where they sometimes continue to pursue normal activities. A chronic phase of clearance begins when deposited particles become sequestered in macrophages of pulmonary connective tissues. These cells are reached by several routes, not least by crossing the bronchial epithelium. Particle clearance from these macrophages is very slow, and residue-containing cells eventually congregate in lymphoid tissues surrounding major bronchi. These findings are discussed as they help to develop an overall picture of clearance from the lungs and as they bear on related topics, such as functional roles of alveolar and pulmonary connective tissue macrophages and the pathogenesis of chronic bronchial disease.     

Tachykinins increase vascular permeability in the gastrointestinal tract of the rat

The occurrence of inflammation as indicated by extravasation of Evans blue bound to plasma proteins was examined in various parts of the gastrointestinal tract in the rat, following administration of tachykinins, capsaicin and hydrochloric acid. Intravenous neurokinin A dose-dependently induced extravasation in stomach, duodenum, jejunum, caecum and colon, but had no effect in ileum. Neurokinin B equipotently induced extravasation in the stomach but had no effect in other parts of the gut and substance P had no effects on extravasation of Evans blue in any of the examined parts of the gastrointestinal tract. Capsaicin given intraperitoneally increased vascular permeability in stomach and duodenum only, while extravasation of Evans blue after capsaicin given intraluminally did not differ from the effect of the vehicle alone. As a comparison, HCl given intraluminally in the duodenum was found to induce a prominent extravasation of Evans blue of a greater magnitude than that of tachykinins. We suggest that tachykinins, and in particular neurokinin A, may be of importance for extravasation of plasma proteins as part of inflammatory reactions in the upper and lower gastrointestinal tract.     

Histopathological study of mucosal and submucosal cysts of stomach.

To clarify the characteristics of mucosal and submucosal gastric cysts, five stomal cancers of the remnant stomach, two cancers with a diffuse cystic distribution and three ordinarily resected cancers were used to study mucinous patterns, proliferative cells and humoral defensive factors in the cystic epithelia. Most cysts showed a gastric appearance with relatively dominant ConA III (+) pyloric gland-like features, and were positively stained with high-iron diamine-alcian blue. Although a few cells positive for proliferative cell nuclear antigen (PCNA) were seen in epithelia, cysts generally revealed a low degree of proliferation, less than 2% of epithelial cells being PCNA (+). Cysts were formed from the isthmus or from ordinary glands, and showed an increase of IgA and secretory component in epithelia of the supranuclear, luminal or basolateral region and increased lysozyme (Ly) activity. Some cysts contained cells with strong positivity for Ly which could not be confirmed as Paneth cells by HE staining. Mucosal and submucosal cysts are considered to arise as a reaction to inflammation, and have a tendency to show both gastric and intestinal characteristics.     

The value of a study of the mucosubstances in rectal biopsies from patients with carcinoma of the rectum and lower sigmoid in the diagnosis of premalignant mucosa

One hundred and twenty-one rectal biopsies from 99 patients with carcinoma of the rectum or lower sigmoid colon were investigated using a high iron-diamine-Alcian blue technique for sulphated and non-sulphated acid mucins. It was found that in normal rectal mucosa sulphomucins are the main carbohydrate component of the goblet cell mucin. This normal ;mucous pattern' changes in the ;transitional' mucosa (histological normal mucosa adjacent to carcinoma) where there is an increase of non-sulphated acid mucins concomitantly with a decrease or absence of sulphated groups in 60 to 90% of the cases according to the type of tumour. The same type of changes in mucus were observed in the ;transitional' mucosa surrounding adenomatous polyps and papillary adenomas; they were not marked in areas around carcinoma in situ and not observed in the metaplastic polyps. These changes seem to be in direct relationship to the grade of differentiation and invasiveness of the tumour.The histochemical changes in the mucins seem to be in favour of a malignant potential in the so-called neoplastic polyps.The high iron diamine-Alcian blue, because of its ;specificity', consistent results, and easy technique is recommended for routine use together with haematoxylin and eosin staining in the diagnosis of premalignant changes.     

Carbohydrate cytochemistry of rhesus monkey tracheal epithelium

Three types of nonciliated secretory epithelial cells contribute material to the mucous lining of pulmonary airways: mucous cells, serous cells, and Clara cells. Extensive interspecies variation exists, especially between humans and laboratory mammals, with regard to occurrence, distribution, and granule content of these secretory cells. This study was designed to characterize one aspect of these differences in one species of nonhuman primate, the rhesus monkey. The complex carbohydrates of secretory granules present in the tracheal epithelium were characterized cytochemically. The tracheas of seven monkeys were fixed by airway infusion, processed, and embedded for both light and transmission electron microscopy. Histochemical stains including Alcian blue-periodic acid Schiff, dialyzed iron, and high iron diamine-Alcian blue were applied to serial methacrylate sections. The mucous cells were the predominant secretory cell type of the trachea and contained periodate-reactive sulfated glycoconjugates. The mucous secretory granules, as resolved with the electron microscope, consisted of a mesh or matrix surrounding a biphasic core. The matrix was stained by all cytochemical reactions used, which included periodic acid-thiocarbohydrazide-silver proteinate, dialyzed iron, low iron diamine, and high iron diamine. The biphasic core also reacted with the four stains, but most intensely with high iron diamine. We conclude from this study that 1) the mucous secretory granule contains carbohydrate throughout all phases of the granule, 2) the mucous granule contains periodate-reactive sulfated glycoconjugates, with sulfate esters concentrated in the core of the granule, and 3) the mucous granules of rhesus trachea morphologically and cytochemically resemble those described in human airways.     

Advanced gastric carcinoma with a complete intestinal metaplasia phenotype associated with early intestinal-type carcinoma

An unusual case of synchronous gastric carcinomas occurred in a 28-year-old man with a family history of gastric disease. Two tumor foci were identified: a well-differentiated advanced carcinoma with the phenotypic properties of complete intestinal metaplasia and an early intestinal-type carcinoma. Histochemical and immunohistochemical stains to demonstrate complete intestinal metaplasia, ie, Alcian blue pH 2.5/periodic acid-Schiff, high iron diamine/Alcian blue pH 2.5, CD10, and MUC2, were all positive in the advanced adenocarcinoma. Of all markers used, only high iron diamine/Alcian blue pH 2.5 and Alcian blue pH 0.5 were positive in the early carcinoma. In these cases, mistakes frequently are made during examination of endoscopic biopsies. Fortunately, the advanced adenocarcinoma was low grade (the patient has shown no signs of disease at 6 years postsurgery). Histopathologic, histochemical, and immunohistochemical findings suggest that an extensive substrate of complete intestinal metaplasia (corpus) and of complete and incomplete intestinal metaplasia (antrum) can be associated with two independent tumors with different phenotypes.