Ulex Europeus agglutinin-I Research Articles

Can domestication shape Canidae brain morphology? The accessory olfactory bulb of the red fox as a case in point

BackgroundThe accessory olfactory bulb (AOB) is the first integrative center of the vomeronasal system (VNS), and the general macroscopic, microscopic, and neurochemical organizational patterns of the AOB differ fundamentally among species. Therefore, the low degree of differentiation observed for the dog AOB is surprising. As the artificial selection pressure exerted on domestic dogs has been suggested to play a key role in the involution of the dog VNS, a wild canid, such as the fox, represents a useful model for studying the hypothetical effects of domestication on the AOB morphology. MethodsA comprehensive histological, lectin-histochemical, and immunohistochemical study of the fox AOB was performed. Anti-Gαo and anti-Gαi2 antibodies were particularly useful, as they label the transduction cascade of the vomeronasal receptor types 1 (V1R) and 2 (V2R), respectively. Other employed antibodies included those against proteins such as microtubule-associated protein 2 (MAP-2), tubulin, glial fibrillary acidic protein, growth-associated protein 43 (GAP-43), olfactory marker protein (OMP), calbindin, and calretinin. ResultsThe cytoarchitecture of the fox AOB showed a clear lamination, with neatly differentiated layers; a highly developed glomerular layer, rich in periglomerular cells; and large inner cell and granular layers. The immunolabeling of Gαi2, OMP, and GAP-43 delineated the outer layers, whereas Gαo and MAP-2 immunolabeling defined the inner layers. MAP-2 characterized the somas of AOB principal cells and their dendritic trees. Anti-calbindin and anti-calretinin antibodies discriminated neural subpopulations in both the mitral-plexiform layer and the granular cell layer, and the lectin Ulex europeus agglutinin I (UEA-I) showed selectivity for the AOB and the vomeronasal nerves. ConclusionThe fox AOB presents unique characteristics and a higher degree of morphological development compared with the dog AOB. The comparatively complex neural basis for semiochemical information processing in the fox compared with that observed in dogs suggests loss of AOB anatomical complexity during the evolutionary history of dogs and opens a new avenue of research for studying the effects of domestication on brain structures.

Carbohydrate profiling of fungal cell wall surface glycoconjugates ofAspergillusspecies in brain and lung tissues using lectin histochemistry

The aim of this study was to evaluate, through lectin histochemistry, the expression of N-acetyl-D-glucosamine, L-fucose, D-galactose and glucose/mannose on the cell wall surfaces of Aspergillus species in histopathological specimens of brain (n = 1) and lung (n = 6) tissues obtained during autopsy of patients diagnosed postmortem as having had invasive aspergillosis. Concanavalin A (Con A), wheat germ agglutinin (WGA), Ulex europeus agglutinin I (UEA-I) and peanut agglutinin (PNA), all conjugated with horseradish peroxidase, were employed. Lectin-binding was visualized using 3,3-diaminobendizine (DAB) and hydrogen peroxide in phosphate buffer solution (PBS). We observed expression of N-acetyl-D-glucosamine and methyl-α-D-mannoside on the cell wall surfaces of all evaluated Aspergillus species, while the expression of L-fucose and D-galactose demonstrated inter and intra-specific variations. The results obtained from this study indicate that the use of WGA and Con A lectins permits visualization of Aspergillus structures such as hyphae, conidial heads and conidia in histopathological specimens of brain and lung tissues.

Carbohydrate profiling of fungal cell wall surface glycoconjugates of Trichophyton tonsurans and other keratinophilic filamentous fungi using lectins

Various researchers have concluded that lectins are useful reagents for the study of fungal cell wall surface glycoconjugates. In this study, we evaluated the expression of N-acetyl-D-glucosamine, L-fucose, D-galactose and glucose/mannose on the cell wall surface of Trichophyton tonsurans and other keratinophilic filamentous fungi, using a simple lectin-binding protocol. The fungal cultures used were isolated from soils obtained from public parks by the hair-bait technique. The lectin assays used concanavalin A (Con A), wheat germ agglutinin (WGA), Ulex europeus agglutinin I (UEA-I) and peanut agglutinin (PNA), all conjugated with horseradish peroxidase. Adhesive tape was placed sticky-side down over the fungal colony, gently pressed and then removed. The fungal-tape samples were incubated with the lectin for 1 h at 4 °C. Lectin binding was visualised using 3,3-diaminobendizine (DAB) and hydrogen peroxidase. There was a high expression of N-acetyl-D-glucosamine on the cell wall surface of all fungi species tested, whereas the expression of L-fucose, D-galactose and glucose/mannose demonstrated inter-specific variations. The lectin-binding assay presented in this article eliminates many of the laborious steps involved in other protocols. The amount and quality of the mycelium and spores immobilised by the adhesive tapes were suitable for obtaining the carbohydrate profile in glycoconjugates of the cell wall surface of filamentous fungi.

Lectins binding during alloxan-induced diabetes in rat soleus muscle

Membrane structural changes of soleus muscle of alloxan-diabetic rats were detected with a panel of six biotinylated lectins. Samples of muscles were obtained from normal and diabetic rats. The biotinylated lectins in staining were detected by avidin-peroxidase complex. Lectin stainning of soleus muscle cryostat sections from alloxan-diabetic rats were compared with similar cryostat sections from normal rats. All lectins were bounded to soleus muscle cell membranes of normal and diabetic rats except the Ulex europeus agglutinin I (UEA-I). UEA-I was not stained for cell membranes of soleus muscles of normal and diabetic rats. But intercellular areas of normal and diabetic soleus muscle were stained for UEA-I. Griffonia simplicifolia I (GS-I) lectin was bounded more strongly to cell membrane of diabetic rats than other lectins. Not only cell membranes but also cytoplasmic miyofibrills of diabetic muscle cells were stained by Griffonia simplicifolia (GS1) lectin. It was concluded that cell membranes of soleus muscle of diabetic rats were differently stained for lectins than those of normal rats. Our data can supplement on the use of lectins as probes for changes in carbohydrate-containing constituents of the alloxan-diabetic muscle cell membrane information obtained from normal and diabetic

Correlation between the sialylation of cell surface Thomsen-Friedenreich antigen and the metastatic potential of colon carcinoma cells in a mouse model.

The cell surface glycosylation profiles of a liver metastatic colon carcinoma variant cell line, SL4 cells previously selected from colon 38 cells in vivo for liver colonization were investigated. Flowcytometric analysis was performed with 7 plant lectins and 10 carbohydrate specific monoclonal antibodies. The results showed that peanut agglutinin (PNA), Sambucus nigra agglutinin, Ulex europeus agglutinin-I, anti-LeX, anti-LeY, and anti-Le(b) antibodies bound to the parental colon 38 cells but not to SL4 cells. Another variant cell line was selected in vitro for the paucity of cell surface PNA-binding sites using a magnetic cell sorter and was designated as 38-N4 cells. The binding profiles of plant lectins and carbohydrate-specific antibodies to 38-N4 cells were very similar to those of SL4 cells. After intrasplenic injections, metastatic ability of 38-N4 cells was higher than that of colon 38 cells. PNA binding to SL4 cells and 38-N4 cells was detected after sialidase treatment of these cells, indicating increased sialylation of Thomsen-Friedenreich antigen in these cells. The mRNA levels of sialyltransferases, ST3Gal I, ST3Gal II, ST6GalNAc I, and ST6GalNAc II, were compared. The level of ST3Gal II mRNA was elevated in both SL4 cells and 38-N4 cells, whereas the level of ST6GalNAc II mRNA was elevated in 38-N4 cells compared with colon 38 cells. According to the expression array analysis, there are other glycosyltransferase genes differentially expressed between SL4 and colon 38 cells, yet their involvement in the altered glycosylation in these cells is unclear.

Differentiation of mitral cell dendrites in the developing main olfactory bulbs of normal and naris-occluded rats

The morphological differentiation of mitral cell dendrites during embryonic and early postnatal development was examined in the main olfactory bulb of rats to determine a possible role of afferent activity in the development of the dendrites. Mitral cells and olfactory nerve fibers were labeled with 1,1`-dioctadecyl-3,3,3`,3`-tetramethylindocarbocyanine perchlorate (DiI) and fluorescein-conjugated lectin (Ulex europeus agglutinin-I), respectively. Morphogenesis of mitral cell dendrites proceeded as previously described (Malun and Brunjes [1996] J. Comp. Neurol. 368:1–16); that is, undifferentiated dendrites with radial orientation were transformed into a single primary dendrite having a glomerular tuft and secondary dendrites extending tangentially into the external plexiform layer. Quantitative examinations in both pre- and postnatal rats revealed that the differentiation of primary dendrites, including tuft formation, increases in diameter and decreases in branching, started before birth, whereas differentiated secondary dendrites were only observed in postnatal animals. Mitral cells with more than two primary dendrites were found after embryonic day 21. The proportion of the mitral cells with differentiated dendrites increased postnatally. At postnatal day 10, almost all mitral cells had fully differentiated dendrites, and mitral cells with multiple primary dendrites were no longer seen. No significant change was found during development in the number of stem dendrites that arose directly from the cell body. Unilateral naris occlusion started on postnatal day 1 retarded differentiation of primary and secondary dendrites, and increased the proportion of mitral cells with multiple primary dendrites. These finding revealed that differentiation of mitral cell primary dendrites precedes that of secondary dendrites, and suggested that the differentiation of secondary dendrites proceeds in an activity-dependent manner. J. Comp. Neurol. 418:402–410, 2000. © 2000 Wiley-Liss, Inc.

Differentiation of mitral cell dendrites in the developing main olfactory bulbs of normal and naris-occluded rats

The morphological differentiation of mitral cell dendrites during embryonic and early postnatal development was examined in the main olfactory bulb of rats to determine a possible role of afferent activity in the development of the dendrites. Mitral cells and olfactory nerve fibers were labeled with 1,1'-dioctadecyl-3,3,3', 3'-tetramethylindocarbocyanine perchlorate (DiI) and fluorescein-conjugated lectin (Ulex europeus agglutinin-I), respectively. Morphogenesis of mitral cell dendrites proceeded as previously described (Malun and Brunjes [1996] J. Comp. Neurol. 368:1-16); that is, undifferentiated dendrites with radial orientation were transformed into a single primary dendrite having a glomerular tuft and secondary dendrites extending tangentially into the external plexiform layer. Quantitative examinations in both pre- and postnatal rats revealed that the differentiation of primary dendrites, including tuft formation, increases in diameter and decreases in branching, started before birth, whereas differentiated secondary dendrites were only observed in postnatal animals. Mitral cells with more than two primary dendrites were found after embryonic day 21. The proportion of the mitral cells with differentiated dendrites increased postnatally. At postnatal day 10, almost all mitral cells had fully differentiated dendrites, and mitral cells with multiple primary dendrites were no longer seen. No significant change was found during development in the number of stem dendrites that arose directly from the cell body. Unilateral naris occlusion started on postnatal day 1 retarded differentiation of primary and secondary dendrites, and increased the proportion of mitral cells with multiple primary dendrites. These finding revealed that differentiation of mitral cell primary dendrites precedes that of secondary dendrites, and suggested that the differentiation of secondary dendrites proceeds in an activity-dependent manner.

Basal layer reactivity in hyperplastic and metaplastic lesions of the bronchi as studied by lectin histochemistry.

The relationship between squamous metaplasia and squamous cell carcinoma of the bronchi has been the subject of controversy. We investigated basal cell hyperplasia, stratification, squamous metaplasia, and squamous cell carcinoma by means of lectin histochemistry with peanut agglutinin (PNA), Ulex europeus agglutinin-I, soybean agglutinin, and Dolichos biflorus agglutinin for all normal basal cells that are reactive. A basal layer stained with PNA was observed in basal cell hyperplasia, stratification, and squamous metaplasia, but this layer was not exhibited by squamous cell carcinoma. In hyperplasia and metaplasia, PNA staining was biased toward the lowest (basal) layer, whereas staining for the other lectins was more uniformly distributed across the layers. A PNA-positive basal layer may be important for the morphologic reversibility of the bronchial mucosa in hyperplastic and metaplastic lesions, and destruction of this layer may be associated with a progression from metaplasia to squamous cell carcinoma.

Detection of blood group differences in human corneal epithelium using a monoclonal antibody and lectins.

To determine if blood group variation in corneal epithelial glycoconjugates can be detected with the use of a monoclonal antibody and sugar-specific lectins. Monoclonal antibodies were developed to isolated corneal epithelial cells using conventional hybridoma techniques. Hybridoma screening and studies of antibody binding to cryostat sections of human corneas of blood groups A, B, and O were performed with immunofluorescence microscopy. Correlation of lectin binding to antibody and blood group binding was done immunohistochemically with lectins conjugated to fluorochromes. Monoclonal antibody designated 1B3.29.36 binds to corneal epithelium of humans of blood group A only. The epitope of the antibody appears to be, in part, N-acetylgalactosamine. The lectins Dolichos biflorus agglutinin (DBA), Griffonia simplicifolia lectin-I isolectin-B4 (GSL-IB4), and Ulex europeus agglutinin-I (UEA-I) bind to epithelium of subjects with blood groups A, B, and O, respectively, and soybean agglutinin binds to all three groups. Variations in corneal epithelium of subjects with different blood groups can be detected with the use of a monoclonal antibody and the lectins DBA, GSL-IB4, and UEA-I. Previous reports of lectin binding to ocular surface epithelium should be reevaluated since they did not take into account blood-group-specific binding.

Assessment of lectin binding for prognosis in endometrial carcinoma

The binding of Ulex europeus agglutinin-I (UEA-I) and Peanut agglutinin (PNA) was studied in 56 endometrial carcinomas. The positive rates of UEA-I and PNA were 66%. Following neuraminidase treatment, the PNA positive rate increased from 66% to 98% and most tumors stained abundantly. UEA-I staining was unaffected. Using a combined grading system based on architectural and nuclear abnormalities, the PNA positive rates of grade 1, grade 2 and grade 3 tumors were 78%, 70% and 36%, respectively. There was no correlation between UEA-I binding and the grading systems. UEA-I binding correlated significantly with the presence of myometrial invasion and vessel permeation. Metastatic endometrial carcinomas showed a high affinity of tumor cells for UEA-I. Tissues that stained with UEA-I were those from patients with the worst prognosis. PNA and UEA-I reactivities relate to the biologic activity of endometrial carcinomas. UEA-I can serve as a prognostic marker for patients with endometrial carcinoma.

Monoclonal antibody GOM-2 binds to blood group B-Le(y) active glycolipid antigens on human gastric cancer cells, KATO-III.

The antigen structure of a mouse monoclonal antibody, GOM-2, established by immunization with KATO-III human gastric cancer cells, was examined. GOM-2 reactive glycolipids were prepared from KATO-III cells and treated with endoglycoceramidase. Structural studies of ten GOM-2 reactive oligosaccharides by a combination of glycosidase digestions, methylation, and affinity chromatography on an Ulex europeus agglutinin I (UEA-I) column revealed that nine of them had a Y-related B-active difucosylated determinant (B-Le(y)) and one had a B-active determinant. Affinity chromatography of the purified and modified oligosaccharides on an immobilized GOM-2 column demonstrated that GOM-2 has a novel binding specificity: it binds tightly to the biantennary structure carrying the B-Le(y) determinant at the termini or the branched structure carrying the B-Le(y) structure at two nonreducing termini.

Immunofluorescent lectin binding patterns and glycoprotein co-localization in the developing murine molar tooth

Fluorescein-conjugated lectins were used in conjunction with antibodies to laminin, tenascin and amelogenin to investigate saccharide expression in the developing tooth germ. At the bud stage, peanut agglutinin (PNA) binding demonstrated residues that may be d-galactose-(β1 → 3) dGalNAc, and this staining occurred after the expression of tenascin. Only the cap-stage enamel organ suprabasal cells and the enamel knot stained intensely with Ulex europeus agglutinin-I, but not Lotus tetragonolobus agglutinin, implying the transient presence of blood group H type I oligosaccharides. At the late stages of amelogenesis, enamel synthesis is preceded by en bloc loss of inner enamel basement membrane components. Before this, Bandeiraea (Griffonia) simplicifolia — I (BSL-I) staining was lost from post-mitotic ameloblasts, suggesting that a glycosylated species is initially removed. Additionally, PNA was co-localized with amelogenin protein, suggesting that it may express β- d-galactosyl sequences. These results indicate that the glycosylation patterns of matrix components during odontogenesis may be important as they vary in a manner similar to that of the well-known glycoproteins.

Interaction of Lectins with Human IgE: IgE-Binding Property and Histamine-Releasing Activity of Twelve Plant Lectins

We examined the IgE-binding reaction and the histamine-releasing response of basophils to a panel of 12 lectins: concanavalin A (Con A), Lens culinaris hemagglutinin (LcH), Pisum sativum agglutinin (PSA), wheat germ agglutinin (WGA), soybean agglutinin (SBA), Bauhinia purpurea agglutinin (BPA), peanut agglutinin (PNA), Ricinus communis agglutinin I (RCA-I), Lotus tetragonolobus agglutinin (Lotus A), Ulex europeus agglutinin I (UEA-I), phytohemagglutinin E (PHA-E) and phytohemagglutinin L (PHA-L), IgE from allergic patients bound with high affinity to Con A, LcH, PSA, RCA-I and PHA-E, and with lower affinity to WGA, BPA, Lotus A and UEA-I, but they did not bind to SBA, PNA or PHA-L. There was no apparent individual difference in the reactivity of IgE to these lectins between 10 IgE preparations from allergic patients. The binding to these lectins, except Lotus A and UEA-I, were competitively inhibited by the lectin-specific sugars or glycopeptide. Upon stimulation by Con A, LcH, PSA, WGA, RCA-1 and PHA-E, leukocytes from allergic patients showed a significant release of histamine, but cells from IgE-deficient subjects did not respond to these lectins. The histamine-releasing responses by these lectins were also inhibited by specific sugars or glycopeptides.

Cytochemistry for Lectins, Actin, Nucleotide Tetrazolium Reductases and Several Phosphatases in the Porcine Semitendinosus Muscle: Vascular Development in Young Pigs

An ontogeny study of the porcine semitendinosus muscle was conducted to meet two objectives: 1) to evaluate enzyme histochemistry, lectin cytochemistry and actin staining for usefulness as quantitative markers of muscle capillaries and 2) to describe the ontogeny of capillary density changes in developing porcine muscle. Muscle samples were obtained from three to eight crossbred pigs at each of the following ages: newborn, 1 to 2 d and 1, 2, 3, 4.5 and 24 wk. Cryostat sections were stained or reacted for alkaline and neutral phosphatase, dehydrogenase, actin, a panel of nine plant lectins (fluorescein isothiocyanate conjugated) and routine cytochemical tests for muscle fiber typing. Capillaries were quantitatively marked by reactions for dehydrogenase activity in the young pigs but not in the oldest animals. The lectins, soybean agglutinin, Bauhinia purpuria agglutinin, Ulex europeus agglutinin-I and Griffonia simplicifolia agglutinin-I (GS-I) quantitatively stained capillaries at all ages. Histological observations of thin sections of epon-embedded tissues served to validate the lectin and cytochemical capillary staining. Micrographs of sections stained with the lectin GS-I were used to count capillaries and muscle fibers so the capillary:fiber ratio (C/F) could be calculated. Deep (red) and superficial (white) aspects of muscle sections had different C/F at birth, 2, 4 and 24 wk. The deep aspects (DA) had higher C/F than superficial (SA) aspects (at all four ages), and there were age-dependent increases (P less than .001) in C/F of DA and SA. This study demonstrates the usefulness of lectin staining for determining C/F in porcine muscle.

Lectin cytochemistry of the dark granules in the type 1 cells of Syrian hamster circumvallate taste buds

Lectin-gold complexes in the dark granules (DGs) and the dense substance (DS) of vallate taste buds were localized. Both the DGs and the DS were labelled with wheat-germ agglutinin, Ulex europeus agglutinin-I and peanut agglutinin. Their common reaction to these lectins suggested that the DGs contain carbohydrate components similar to those of the DS. The results provide cytochemical evidence that the DS in the taste pit represents, at least in part, the content of the DGs in the type 1 cells.

Lectin histochemistry of mammalian endothelium.

Lectin-histochemical studies were performed on formalin-fixed, paraffin-embedded tissues from ten mammalian species to demonstrate the pattern of carbohydrate residues in vascular endothelium. Ten different biotinylated lectins were used as probes and avidin-biotin-peroxidase complex (ABC) was used as visualant. Ricinus communis agglutinin-I (RCA-I) and wheat germ agglutinin (WGA) stained vascular endothelium in all species. Peanut agglutinin (PNA) stained vascular endothelium in all species only after preincubation with neuraminidase. Bandeirea simplicifolia agglutinin-I (BS-I) stained vascular endothelium in all species but human, while Ulex europeus agglutinin-I (UEA-I) stained only human endothelium. Individual differences in staining of human vascular endothelium were noted with BS-I and succinylated-WGA (SWGA). Similarly, individual differences in staining of animal vascular endothelium were noted with soybean agglutinin (SBA) after preincubation with neuraminidase. Finally, Concanavalia ensiformis agglutinin (Con A), Dolichos biflorus agglutinin (DBA) and Lens culinaris agglutinin (LCA) did not stain vascular endothelium in any of the species studied.

MUCOSUBSTANCE HISTOCHEMISTRY OF THE NORMAL MUCOSA AND EPITHELIAL NEOPLASMS OF THE LUNG

Histochemical properties of mucosubstances in normal lung tissue and carcinoma of 30 operated cases were investigated using various histochemical methods. Epithelial cells lining the respiratory tract were characterized by their specific mucosubstances. Namely, the ciliated cells had a thick surface coat consisting of sulfomucins. Basal cells showed intense reactivity to Ulex europeus agglutinin-I (UEA-I) and Arachis hypogaea agglutinin (PNA). The goblet cells differed from the glandular mucous cells in their PNA and UEA-I reactivities. The Clara cells and type II pneumocytes resembled each other in having a surface coat consisting of PNA- and Maclura pomifera agglutinin (MPA)-reactive sialomucins. Carcinoma cells also had mucosubstances varying with cases and with cells of the same case, but surface coat-type mucin of Clara cell-type adenocarcinoma resembled that of the Clara cell and type II pneumocyte. Squamous cell carcinoma had only a small amount of mucosubstance which was histochemically similar to that of the basal cell.

上気道粘膜における糖たん白の組織化学的研究 正常声帯と声帯ポリープについて

Histochemical studies of the membranous glycoprotein with horse-raddish peroxidase conjugated lectins were performed on 3 cases of normal laryngeal mucosa, which were obtained operativelly from the patients with hypopharyngeal cancer, and on 13 cases of vocal cord polyps extirpated micro-laryngo-surgery.The results obtained were as follows:1) In stratified squamous epithelium of normal vocal cord, basal and keratinezed stratum differed from the middle stratum lacking in affinity for Ulex europeus agglutinin-I (UEA-I). And the staining with peanut agglutinin (PNA) showed specific binding to the spinous and lower granular layer.2) In stratified squamous epithelium of vocal cord polyps, they were classified into the following three types according to thier staining features with both lectins. a) UEA-I binded strongly to the upper spinous and granular cells. Some cells of the granular layer showed variable intensity ranging from lightly to strongly positive staining with PNA. Both of edematous and fibrous changes were observed in the stroma of these polyps. b) All cell layers except for basal layer were stained with UEA-I. The spinous layer and some cells of the granular layer were stained with PNA. In these polyps, edematous stroma was observed. c) Inter-cellular staining of the lower spinous layer showed specific feature to UEA-I. The basal and lower spinous layers were stained with PNA. Fibrin precipitation, hemorrhage and plasma infiltration were shown in the stroma of these polyps.These data may strongly suggest that expression of membranous glycoprotein in epithelium differs according to the maturation of cells from the basal cell to the more mature keratinocyte, and also indicate that lectins may be used as marker in various stages of normal and abnormal differentiation.

Ultracytochemistry of glycosylation sites of rat colonic mucous cells with labeled lectins.

The mucous glycoproteins of mucous cells in the lower half of the rat proximal colon were examined ultracytochemically to study the intracellular sites of glycosylation in relation to the cell differentiation and the functional polarity of the cell organellae using a combination of a hydrophilic resin embedment and a postembedding staining technique with lectin gold com plexes.In the immature mucous cells located near the branching point of the glandular crypt, the Golgi stacks and mucous granules were stained with soybean agglutinin (SBA), Ricinus communis agglutinin-I (RCA-I) and Helix pomatia agglutinin (HPA). In the mature mucous cells at the bottom of the crypt, the cis region of the Golgi stack was stained with SBA, a few intermediate saccules were stained with RCA-I, and both the cis and trans sides of the cisternae and mucous granules, but not the intermediate lamellae, were stained with HPA. After neuraminidase digestion of ultrathin sections, the RCA-I binding sites were greatly increased. Ulex europeus agglutinin-I (UEA-I) and Limax flavus agglutinin (LFA) positively stained the trans side lamellae and mucous granules of the mucous cell at the bottom of the glandular crypt.The processing of the glycosylation of the mucous glycoprotein in the lower half crypt of the rat proximal colon was partly clarified electron microscopically.

ABH Blood Group Isoantigen Expression in Breast Carcinomas—An Immunohistochemical Evaluation Using Monoclonal Antibodies

The ABH blood group isoantigen status of a retrospective series of 233 invasive breast carcinomas was examined, employing monoclonal antibodies (MCAB) against A, B, and H antigens with the avidin-biotin-peroxidase complex method. In addition, the H antigen was localized with Ulex Europeus Agglutinin I (UEAI) binding. The MCABs provided consistent and specific staining of erythrocytes and endothelium, as well as normal and neoplastic breast epithelium. The anti-H MCAB yielded cleaner background and less intense staining, but otherwise the staining distribution was comparable to the UEA I technique. Contrary to previous reports, deletion of isoantigen expression was not universal in all invasive carcinomas. Whereas 64%, 77%, and 73% of carcinomas from groups A, B, and AB patients, respectively, demonstrated total isoantigen loss, the remaining tumors exhibited variable degrees of isoantigen expression. Moreover, those carcinomas with complete loss of A and B determinants still displayed variable degrees of H immunoreactivity. Carcinomas from group O patients had different degrees of H antigen deletion, with only 12% showing total loss. Statistical analysis revealed that the isoantigen status bore no significant relationship to various epidemiologic, clinical, and pathologic parameters and did not serve as a useful prognostic determinant.