The epithelia of the protrusible tongue of Eurycea longicauda guttolineata (Hoolbrook 1838) (Urodela: Plethodontidae)

In this study the lingual and sublingual glands, the lingual stem and the epithelial surface of the protrusible secondary tongue were investigated by light, scanning and transmission electron microscopy. The quality of the secretions of the epithelia was characterized histochemically. The lingual epithelium is formed by superficial (pavement) and goblet cells and at the margin of the tongue pad are also regions covered by ciliated cells. On the dorsal part of the tongue there are goblet cells of type A with mainly acidic secretions and of type B containing neutral secretions. Most of the goblet cells on the ventral side of the tongue (hypoglottis) show a strong alcian blue/PAS positive reaction (type I) and some produce neutral secretions (type II). The glandular cells of the lingual gland react positively to alcian blue and PAS in the apical region of the gland. In contrast there is only alcian blue-positive staining in the basal part of the gland. The size and complexity of the inclusion bodies of the secretory granules increase in a basal direction. In addition, there are ciliated cells in the glandular epithelium. Although the epithelium of the lingual stem is thin, it is double-layered. The cell types observed in this region are identical to those of the ventral part of the protrusible tongue. At the margin of the sublingual gland are trough-like structures. In the center, tubular parts are observed. The cells of this gland are stain strongly with alcian blue (pH 1.0) mainly in the basal part of the gland. The results of this are compared to the tongue pad and the lingual gland of Salamandra salamandra and Ambystoma mexicanum.     

Surface secretions of the skin of Blennius (Lipophrys) pholis L.

Mucus is secreted to the surface of the body and fin webs of Blennius pholis by superficial epithelial cells and by goblet cells. Some goblet cells secrete sulphated acid glycoproteins, others produce a mucus which is neutral or mixed in its reactions. The superficial epithelial cells of these areas secrete sulphated acid glycoproteins, seen by electron microscopy as electron-lucent or moderately lucent vesicles; this secretion is not normally visible external to the skin in transmission electron microscope (TEM) sections. These cells do not react to the bromphenol blue test for proteins. Over part of the surface of the pelvic fins and the distal parts of the rays of the pectoral fins, the skin contains no goblet cells and bears a thick external secretion, or cuticle, containing protein and glycoprotein which is mainly neutral in reaction, although some cells at the edges of the region secrete weakly sulphated or non-sulphated acidic glycoprotein. The protein content of the columnar superficial epithelial cells of these regions correlates with the fibrous nature of the secreted cuticular layer as seen by TEM; the columnar cells are characterized by extensive ribosomal endoplasmic reticulum and vesicles which stain darkly with phosphotungstic acid, less so with uranyl acetate. The distal part of the cell, containing these vesicles, reacts positively to the PAS stain. In some places the borders of the zones with fibrous cuticle are characterized by cuboidal superficial epithelial cells which give a strong positive reaction to alcian blue at pH 1.0, indicating the presence of sulphated acid glycoproteins, but also react positively to the bromphenol blue test for proteins.     

Histochemistry of small intestinal dysplasia in familial polyposis coli

Biopsies of duodenal and ileal mucosa from patients with familial polyposis coli were studied. Areas of atypia were identified in the duodenum of six patients and in the ileum of three patients. Grade I atypia was characterized by crowding and elongation of cells and nuclei, a slight reduction in the number of goblet cells and the presence of a brush border; grade II atypia was further characterized by pseudo- or pluristratification of cells, a marked reduction in the number of goblet cells and the absence of a brush border. In areas of atypia, columnar cells often contained PAS-positive apical granules, which were diastase-resistant and unstained by alcian blue at any pH; the brush border, even where recognizable in haematoxylin-eosin and PAS-stained sections, was unreactive histochemically for alkaline phosphatase. Goblet cells were few in areas of atypia, but those present were regularly stained by PAS and alcian blue pH 2.6. Apical granules, similar in their histochemical characteristics to those observed in columnar cells in areas of atypia, were also found in otherwise normal mucosal areas, even in some patients with no overt areas of atypia in the biopsies studied. These granules have been interpreted as an abnormality, possibly preceding the onset of atypia. Hyperplasia of goblet cells, secreting mucins with the same staining pattern as in normal intestine, was found in some patients, either adjacent to areas of atypia or independent of them. Intervening columnar cells had a normal morphology, alkaline phosphatase-reactive brush borders and no sign of mucus secretion. This goblet cell hyperplasia has been interpreted as a reactive, nonspecific alteration of the mucosa.     

Some histochemical reactions of mast cells of gerbils,hogs, armadillos and cats

Summary Mast cells of the Mongolian gerbil Meriones unguiculatus, the hog Sus scrofa, the cat Felis catus and the armadillo Pasypus novemcinctus were studied histochemically in relation to various fixation procedures, using azure A at pH 1 and 3, alcian blue at pH 1 and 2.5, diazosafranin at pH 3 and 7.8–8, and the PAS reaction. Fixations were performed in buffered 10% formol and 5% glutaraldehyde, in Kose's fluid, buffered sublimate (B4), lead nitrate and lead acetate formol.With azure A and alcian blue many mast cells were found in the gerbil with the aldehyde fixatives, fewer with the heavy metals. The diazosafranin reaction was present only in the aldehyde material, the PAS reaction was negative.In the hog, mast cells were more numerous after heavy metal fixation, fewer with aldehydes. Azure A stained metachromatically at pH 1 and 3, alcian blue reacted only at pH 1, the PAS reaction was negative, the pH 3 and 8 diazosafranin reactions were positive with all 4 fixations.In the cat, mast cells were moderately numerous with lead acetate formol, rare with formol and absent with glutaraldehyde. They stained with azure A at pH 1 and 3, with alcian blue at pH 1 and 2.5, with diazosafranin at pH 3 and 8 and by the PAS reaction.Armadillo mast cells were more numerous after heavy metal fixations, stained with azure A and alcian blue at pH 1 and 2.5 to 3, and with acid and alkaline diazosafranin.The mast cells of the 4 species vary in their requirements for aldehyde and heavy metal fixation, in their PAS reactivity and in their pH 2.5 alcian blue staining. All are sufficiently sulfated to react to cationic dyes at pH 1, but vary in PAS reactivity, indicating partial or complete sulfation. The presence of 5-hydroxytryptamine is indicated in all four species.Assisted by grant from National Cancer Institute C-04816.     

Use of an improved zirconyl hematoxylin stain in the diagnosis of Barrett's esophagus

Barrett's esophagus is a precancerous condition characterized by replacement of the normal stratified squamous epithelium by a simple columnar epithelium with goblet cells that secrete an acidic mucin. As originally formulated, fresh solutions of zirconyl hematoxylin stain goblet cells poorly. An improved formula, quintupling the amount of oxidant, yields zirconyl hematoxylin solutions that stain goblet cells darkly even when fresh. The improved zirconyl hematoxylin can be used in place of alcian blue in the diagnosis of Barrett's esophagus. The ingredients of zirconyl hematoxylin are always readily available and are generally recognized as safe.     

Histological and morphological evaluations of the digestive tract and associated organs of haddock throughout post-hatching ontogeny

At hatching, the oesophagus of haddock Melanogrammus aeglefinus lacks goblet cells, the intestine is a simple undifferentiated tube, the liver is present as a rounded mass caudal to the heart, and numerous zymogen granules are present in the pancreas. The first intestinal convolution appears at day 2, at the posterior end of the digestive tract. The oesophagus displays alcian blue and PAS positive mucus secreting cells on day 12, which become numerous by day 15. By day 18, epithelial cells of the posterior intestine show evidence of protein absorption in the form of supranuclear vacuoles. The swimbladder inflates in 50% of the larvae by day 22, although inflation rate is highly variable. By day 35, or 10 mm, a pyloric caecal ridge appears which separates the presumptive stomach, which is now showing evidence of gastric gland formation, from the intestine. This marks the beginning of digestive features characteristic of the juvenile stage.     

Histochemical features of the Muscovy duck small intestine during development

We demonstrated for the first time the distribution and morphology of argyrophil and of goblet cells in the mucosa of the small intestine of the Muscovy duck during development using the Grimelius silver staining and alcian blue/periodic acid-Schiff (AB/PAS) staining technique. The argyrophil cells distribution was variable over the length of the small intestine from embryonic day 24 (24E) to post-hatching day 13 (13d). In the villi most argyrophil cells belonged to the open-type, while in the crypts they belonged to the closed-type. In the duodenum the density of argyrophil cells was highest at hatching, while in the jejunum and in the ileum the highest density value was at hatching and 13d. AB/PAS-positive goblet cells appeared on the villi and crypts of the duodenum and jejunum at 30E, and in the ileum at hatching. The density of AB/PAS-positive cells was the highest in the three segments at hatching. The AB-positive cells, compared with the PAS-positive cells, predominated in villi and crypts of the three segments, moreover the rate of AB-positive cells to PAS-positive cells significantly decreased from 30E to 9d. An increase in argyrophil and goblet cells number during the later incubation and at hatching, could indicate the small intestine in that period is being prepared to face a new diet.     

Zirconyl Hematoxylin: a New Stain for Acidic Mucins

Most stains for acidic mucins are time-consuming to prepare and have poor stability. Zirconyl hematoxylin is easily prepared and works for a year or more. It is made by adding 5 ml freshly-made 0.1% aqueous sodium iodate, 400 mg zirconyl chloride oc-tahydrate, and 40 ml 25% aqueous glycerol, in that order, to 100 mg of hematoxylin in 5 ml of absolute ethanol and stirring for 5 min. Stain 10 min and do not “blue” the stain. Chlorazole black or kernechtrot and fast green are good counterstains. Zirconyl hematoxylin stains acidic mucins violet or red violet, regardless of how they are fixed. It stains the same mucins as alcian blue in mouse and sheep salivary glands. It shows goblet cells in mouse rectum as well as alcian blue. It stains the same stomach regions in a lizard as alcian blue. Like alcian blue and colloidal iron, zirconyl hematoxylin stains the mucin of cancerous prostate, but not normal prostate.     

小肠上皮分泌细胞特性的组织学观察

目的寻找一种可以替代人体消化管的动物标本,并通过特殊染色方法,使得小肠上皮分泌细胞的形态特征能够明显地显示出来。方法随机采集成年猫小肠的新鲜标本,经Bouin液灌注固定24h后,石蜡包埋切片脱蜡入水。分别采用Gomori染色法、PAS反应、Gomori＋PAS反应、阿利新蓝（alcian blue,AB）染色法、AB＋PAS反应、HE染色法和苏木精-焰红染色法进行染色。结果在各种染色的切片标本上,能够观察到杯状细胞的形态、分布和染色特性以及肠内分泌细胞的特点,并发现在它们之间还存在一种绿色颗粒细胞和嗜酸性颗粒细胞。结论通过特殊染色可以肯定猫的小肠杯状细胞合成的是中性粘蛋白和酸性粘蛋白;绿色颗粒细胞为未成熟杯状细胞;嗜酸性颗粒细胞为Paneth细胞,其特点是单个分散分布。肠内分泌细胞与周围其他上皮细胞的染色对比明显而容易识别。     

Zirconyl Hematoxylin: a New Stain for Acidic Mucins

Most stains for acidic mucins are time-consuming to prepare and have poor stability. Zirconyl hematoxylin is easily prepared and works for a year or more. It is made by adding 5 ml freshly-made 0.1% aqueous sodium iodate, 400 mg zirconyl chloride oc-tahydrate, and 40 ml 25% aqueous glycerol, in that order, to 100 mg of hematoxylin in 5 ml of absolute ethanol and stirring for 5 min. Stain 10 min and do not “blue” the stain. Chlorazole black or kernechtrot and fast green are good counterstains. Zirconyl hematoxylin stains acidic mucins violet or red violet, regardless of how they are fixed. It stains the same mucins as alcian blue in mouse and sheep salivary glands. It shows goblet cells in mouse rectum as well as alcian blue. It stains the same stomach regions in a lizard as alcian blue. Like alcian blue and colloidal iron, zirconyl hematoxylin stains the mucin of cancerous prostate, but not normal prostate.     

南方鲇消化道杯状细胞分布及类型探讨

本研究应用阿利新蓝－过碘酸雪夫氏（ＡＢ－ＰＡＳ）染色方法对南方鲇（Ｓｉｌｕｒｕｓ　ｍｅｒｉｄｉｏｎａｌｉｓ）消化道杯状细胞的分布特点进行了分类研究,结果表明：南方鲇消化道杯状细胞的数量从肠前段、肠中段到肠后段,呈现递增趋势,有杯状、囊状和梨形三种不同的形态,ＡＢ反应阳性,ＰＡＳ反应阴性,分泌物主要含酸性粘多糖,属Ⅱ型粘液细胞。     

Histochemical and Biometric Study of the Gastrointestinal System of Hyla Orientalis (Bedriaga, 1890) (Anura,Hylidae)

This study was carried out to assess the localization of hyaluronic acid (HA) and the distribution of glycoproteins in the gastrointestinal system of adult Hyla orientalis. Histochemical analysis of the gastrointestinal system in H. orientalis showed that mucous content included glycogene and/or oxidable dioles [periodic acid/Schiff (PAS)+], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS+) and acid sulphate [Aldehyde fuchsin (AF)+] glycoproteins. However the mucus content was not the same in stomach, small and large intestine. The mucus content of stomach included only glycogene and/or oxidable dioles and sialic acid residues. Besides these histochemical methods, the localization of HA was detected using biotinylated hyaluronic acid binding protein labeled with streptavidin-fluorescein isothiocyanate (FITC). In the extracellular matrix of the submucosa, the reaction for HA was evident. Since HA was located in submucosa beneath the epithelial layer of gastrointestinal system, it has a significant role in hydric balance, and essential to provide the gastrointestinal system integrity and functionality. According to biometric results, there were statistical differences between small and large intestine in terms of the amount of material stained positive with PAS/AB, PAS, KOH/PAS and AF/AB. Additionally, number of goblet cells in the small and large intestine was significantly different.Key words: Gastrointestinal system, goblet cell, glycoproteins, hyaluronic acid, amphibian, Hyla.     

Studies on silk secretion in the trichoptera (F. Limnephilidae): I. Histology,histochemistry, and ultrastructure of the silk glands

As part of a study on trichopteran silk secretion, the histology, histochemistry, and ultrastructure of the silk glands of two species of limnephilid trichopteran larvae, Pycnopsyche guttifer (Walk.) and Neophylax concinnus McL., were investigated. The silk glands consist of three anatomically distinct regions: a long, posterior silk-secreting region; a shorter, anterior conducting tube; and a terminal press/common duct. In Pycnopsyche, there is also a modified bulbous region between the secreting and conducting areas. Each anatomical region has a distinct cell type. There are two structurally and histochemically different components of the secretion in the glandular lumen: a core and a peripheral layer. Both components are produced all along the gland and are principally proteinaceous. However, the peripheral layer is also PAS and alcian blue (pH. 2.5) positive and shows β-metachromasia with toluidine blue (pH 3.5), indicating the presence of both neutral and acidic polysaccharides.     

Morphological Descriptions of the Esophagus of the Sea Bream (<Emphasis Type="Italic">Sparus aurata</Emphasis>, Linneaus 1758)

The current work prepared to describe the microscopic morphological appearance of the esophagus of the gilthead sea bream (Sparus aurata). The esophageal wall was organized into four tunica; mucosa, submucosa, muscularis and serosa. There are many goblets cells in the esophageal epithelium. The tunica muscularis is very thick and consisting of; an outer circular thick layer and an inner longitudinal layer of striated muscle. The presences of the many goblet cells help in the esophageal lubrication and protection. By the histochemical reactions with Alcian blue and PAS staining, the esopgaster secretory epithelium devoid from any goblet cells. There are many mucous droplets of the high electron density and the low electron density between surface epithelial cells within the goblet cells. The obtained results of the current investigation may help to understanding the physiology of the esophageal digestive functions in Sparus aurata.     

Effects of novel (Streptomyces) hyaluronidase digestion upon some mucosaccharide stainings of the cartilages and aortas in the rabbit and rat

Summary Effects of novel (Streptomyces) hyaluronidase digestion upon the alcian blue (pH 1.0), alcian blue (pH 2.5), periodic acid-Schiff (PAS) and alcian blue-PAS stainings of mucopolysaccharides have been tested in the cartilage and aorta tissues of the rabbit and rat. These effects were compared with those of testicular hyaluronidase digestion upon the same stainings in the tissues. Evidence has been presented that a Streptomyces hyaluronidase digestible alcian blue (pH 2.5) reactive mucosubstance (hyaluronic acid) is present in the peripheral areas of the cartilage tissues and all the layers of the aorta tissues. Furthermore, the histochemical data obtained in this study appear to present a base line for establishing a promising enzyme digestion technique for the identification of hyaluronic acid in mucosaccharide histochemistry.     

Reproductive cycles of Mugil cephalus, Liza ramada and Liza aurata (Teleostei: Mugilidae)

Carbohydrates in photoreceptor segments in the retina of the firemouth cichlid Thorichthys meeki are described and compared. The present periodic acid-Schiff (PAS) and lectin results revealed the occurrence of neutral carbohydrates, composed mainly of glycosamine and mannose and glucose, in the light absorbing segments in rods and cones in this species. Unlike in mammals, the retina in this teleost seems to be poor in galactose and galactose-galactosamine units in the light absorbing segments.     

A method for the identification of sulfated glycopeptide by two-dimensional electrophoresis on cellulose acetate membrane

Two-dimensional electrophoresis on cellulose acetate membrane permits the clean separation of sulfated glycopeptide in a mixture of acidic glycans (glycosaminoglycans and acidic glycopeptides). Two systems were used. In system 1, 0.1 M pyridine-0.47 M formic acid buffer (pH 3.0) was used in the first and 0.1 M barium acetate (pH 8.0) in the second dimension. In system 2, 0.1 M pyridine-0.47 M formic acid buffer (pH 3.0) was used in the first and 0.1 M HCl in the second dimension. All of the acidic glycans on electrophoretogram were stained with alcian blue in 70% ethanol. On the other hand, sulfated glycans alone were made visible with alcian blue in 0.1 M HCl. Alcian blue in 70% ethanol or 0.1 M HCl, when combined with periodic acid-Schiff's reagent identified sulfated glycopeptides on cellulose acetate membrane.     

Effect of nutrition and Enteromyxum leei infection on gilthead sea bream Sparus aurata intestinal carbohydrate distribution

The effect of a practical plant protein-based diet containing vegetable oils (VO) as the major lipid source on the mucosal carbohydrate pattern of the intestine was studied in gilthead sea bream Sparus aurata challenged with the myxosporean parasite Enteromyxum leei. Fish fed for 9 mo either a fish oil (FO) diet or a blend of VO at 66% of replacement (66VO diet) were exposed to parasite-contaminated water effluent. Samples of the anterior, middle and posterior intestine (AI, MI and PI, respectively) were obtained for parasite diagnosis and histochemistry. Fish were categorised as control (C, not exposed), early (E) or late (L) infected. Mucin and lectin histochemistry was applied to detect the different types of mucins and sialic acid in goblet cells (GC), the brush border and enterocytes. The number of GC stained with periodic acid Schiff (PAS), alcian blue (AB), aldehyde fuchsin-alcian blue (AF-AB), for the detection of neutral, acidic, sulphated and carboxylic mucins, and with the lectin Sambucus nigra agglutinin (SNA), were counted in digital images. The 66VO diet produced a significant decrease of GC with neutral and acidic mucins in the AI and MI, and also of those with carboxylic mucins and sialic acid in the MI. Sulphated mucins and sialic acid were less abundant in the AI than in the MI and PI in the C-66VO treatment. E. leei infection had a strong effect on the number of GC, as E and L infected fish had a significant decrease of GC positive for all the stains versus C fish in PI. Time and diet effects were also observed, since the lowest values were mostly registered in E-66VO fish in PI. In conclusion, though GC depletion was mainly induced by enteromyxosis, an effect of the diet was also observed. Thus, the diet can be a predisposing factor that worsens the disease course.     

Expression of trefoil peptides in the subtypes of intestinal metaplasia

We studied the expression of trefoil peptides in the different types of intestinal metaplasia of the stomach. Endoscopic biopsy was performed in 132 patients with dyspepsia. Intestinal metaplasia subtype was classified according to the pattern of alcian blue/PAS staining and high iron diamine staining. Expression of trefoil peptides was measured by immunohistochemistry. TFF1 and TFF3 were mainly expressed in goblet cells and TFF2 in columnar cells in all the types of intestinal metaplasia. There was a gradual decrease of TFF1 and TFF3, and increase of TFF2, during the progression of intestinal metaplasia from type I to type III via the type II intermediate.