Ageing of the human oviduct: lectin histochemistry.

The aim of the present research was to investigate the changes of the sugar residues in the oviduct in the course of ageing in postmenopausal women vs normally menstruating women, by means of lectin histochemistry. Twenty asymptomatic postmenopausal women (48-83 years old) were recruited among patients who underwent a vaginal hysterectomy. Eight normally menstruating women were recruited as controls. Fragments of Fallopian tubes (pars ampullaris) were fixed in 10% formalin and routinely processed. The sections were labelled with HRP-lectins (PNA, SBA, DBA, WGA, Con A, LTA, UEAI). Some sections were pre-treated with neuraminidase prior to staining with HRP-lectins. Among the postmenopausal patients, our histochemical data showed that there was no difference in the localization and distribution of sugar residues of glycoconjugates as detected by various HRP-lectins. Moreover, our results demonstrated that the oviductal epithelium is characterized by apical reactivity in both ciliated and non-ciliated cells. In the course of ageing, the ciliated cells changed their morphology from bathyprismatic to large and rounded shape. ConA lectin reacted intensely with such highly degenerating ciliated cells and could be considered a marker of these cells. The degenerating ciliated cells are also characterized by the absence of sialic acid. In comparison with the sugar residues present in the control group, the oviductal epithelium of postmenopausal women is characterized by the loss of reactivity with DBA, WGA and ConA. Moreover, PNA reactive material was present at the free border of the ciliated and non-ciliated cells. The latter findings were statistically confirmed and could be considered strictly related to the ageing process.

Glycoconjugate sugar residues in the chick embryo developing lung: a lectin histochemical study.

A lectin histochemical study was performed to investigate the distribution and changes of the oligosaccharidic component of the glycoconjugates in the lung of chick embryos, of 1-day-old chick, and of the adult animal. For this purpose, a battery of seven horseradish peroxidase-conjugated lectins (PNA, SBA, DBA, WGA, Con A, LTA, and UEA I) were employed. During the first phase of parabronchi and atria formation, D-galactose-(beta1-->3)-N-acetyl-D-galactosamine, beta-N-acetyl-D-galactosamine, D-glucosamine, alpha-D-mannose, and sialic acid, present at the level of the surface and of cytoplasmic granules of the lining epithelial cells, seem to play a role in regulating morphogenetic phenomena. In the subsequent phases, the parabronchial lumen and the atrial cavities were characterized by the presence of lectin-reactive material rich in terminal D-galactose-(beta1-->3)-N-acetyl-D-galactosamine, beta-N-acetyl-D-galactosamine, D-glucosamine and alpha-D-mannose. From day 18 onwards and immediately after hatching, the free border of the cells lining the air capillaries was characterized by the presence of beta-N-acetyl-D-galactosamine and alpha-D-mannose. The appearance of these sugar residues was concomitant with the beginning of respiratory activity.

Lectin histochemistry of goblet cell sugar residues in the gut of the chick embryo and of the newborn.

The anlage of duodenum, ileum and colon were removed from chick embryos of day 8-21 of incubation and from 1-day-old chicks. A battery of seven different horseradish peroxidase-conjugated lectins (PNA, SBA, DBA, Con A, WGA, LTA and UEAI) was used to study the carbohydrate residues of the glycoconjugates in the goblet cells of the three parts of the intestine. The main results can be summarized as follows: differences in lectin binding were absent in the proximal and distal parts of the duodenum, ileum and colon. Lectin histochemistry showed differences among the three intestinal segments for the time of appearance of the oligosaccharides in the goblet mucus. In the colonic goblet cells of 1-day-old chicks, LTA and UEAI lectins showed two different types of linkage of alpha-L-fucose. This is the first demonstration of UEAI reactive sites in Gallus domesticus.

The oligosaccharidic component of the glycoconjugates in lichen planus, granuloma annulare, seborrheic keratosis and plamoplantar keratoderma: lectin histochemical study.

It is well known that cell surface glycoconjugates play an important role in cell proliferation, adhesion and differentiation. The aim of this investigation was to define the changes of the glycoconjugate saccharidic moieties in the epidermis and derma of patients affected by several skin pathologies such as seborrheic keratosis, lichen planus, granuloma annulare and palmoplantaris keratoderma. Bioptical specimens from skin lesions as well as from normal skin were fixed in Carnoy's fluid and routinely processed. The sections were treated with HRP-lectins (PNA, DBA, SBA, WGA, ConA, LTA and UEAI). Cytochemical controls were performed for specificity of lectin-sugar reaction. Some sections were pre-treated with neuraminidase prior to staining with HRP lectins. In comparison with normal human skin, epidermal lectin binding pattern in the considered diseases showed considerable qualitative and quantitative variations. In general, in all the considered pathologies, a lack and/or a decrease in lectin binding at the epidermal layers was observed; among the various diseases, differences in cellular localisation of the sugar residues were also noted. In such respect, an exception was represented by seborrheic keratosis, where the cells of the basal layer showed PNA reactivity, which was absent in the basal layer of the normal skin. Although seborrheic keratosis and lichen planus have been studied by others authors, our findings are not in total accordance concerning lectin binding; this is probably due to the different fixatives employed. Our findings seem to reveal significant changes in keratinocyte glycoconjugate oligosaccharides in the previously mentioned diseases, providing clues to their pathogenesis.

Lectin binding in the human endometrium in early luteal phase following controlled ovarian hyperstimulation.

A lectin histochemical study was performed to investigate the glycoconjugate saccharidic moieties on the endometrial epithelium and stroma in 12 women undergoing controlled ovarian hyperstimulation (COH) for in-vitro fertilisation for embryo transfer (IVF-ET) in early luteal phase. 7 control subjects were also evaluated. For this purpose a battery of seven horseradish peroxidase-conjugated lectins (PNA, SBA, DBA, WGA, ConA, LTA and UEA I) was used. Cytochemical controls were performed for specificity of lectin-sugar reaction. As far as the endometrial glands and stroma are concerned, the obtained data showed no differences in the endometrial lectin binding between the subjects of the control group and the ones undergoing COH, with the exception of PNA reactivity at the level of the apical portion of the glandular cells, which was detected only in COH women. It is noteworthy that, although the endometrial dating using the Noyes's criteria showed marked dissynchronies between the stroma and the glands in COH subjects, a uniformity of lectin binding, revealing the same type and localization of terminal oligosaccharides, was observed in all the examined subjects. The uniformity in distribution of the sugar residues detected in the endometrial specimens following COH might be due to the massive FSH and/or hCG treatment which probably determines an endometrial environment almost equal in all the examined subjects. In all the treated subjects reactivity with sialidase-WGA and ConA, revealing the presence of N-acetyl-D-glucosamine and D-mannose respectively, was detected at the level of the lining epithelium.

Sugar residues content and distribution in atrophic and hyperplastic postmenopausal human endometrium: lectin histochemistry.

A lectin histochemical study was performed to investigate the glycoconjugate saccharidic moieties of the human postmenopausal endometrium (14 atrophic and 15 hyperplastic). For this purpose a battery of seven horseradish peroxidase-conjugated lectins (PNA, SBA, DBA, WGA, ConA, LTA and UEA I) was used. No differences in lectin binding between atrophic and hyperplastic endometria were observed. This investigation allowed us to provide a basic picture of the oligosaccharidic distribution in postmenopausal endometria. The data on the saccharidic distribution at the postmenopausal endometria showed a large amount of sugar residues at all the investigated sites, i.e. the lining and glandular epithelium, the stroma and the vessels (capillary and large vessels). Furthermore, at the endometrial lining epithelium, at the glands and at the wall of the blood vessels of some postmenopausal women the presence of alpha-L-fucosyl residues which bind via alpha (1-6) linkage to penultimate glucosaminyl residues and/or difucosylated oligosaccharides was demonstrated for the first time.

Aging of the human endometrium: peri-implantation phase endometrium does not show any age-dependent variation in lectin binding.

OBJECTIVE: To evaluate if the peri-implantation endometrium shows age variations in lectin patterns, which suggest possible age variations in embryo-maternal recognition. STUDY DESIGN: Peri-implantation endometria of younger ( < 30 years of age: n = 13) and older ( > 40 years of age: n = 17) normally menstruating women was studied. Endometrial specimens were routinely fixed in buffered formaline and embedded in paraffin. Sections (5 microns) were studied using seven lectins: DBA (Dolicus biflorus, binding specificity alpha-D-GalNAc), PNA (Arachis hypogea, binding specificity D-Gal (beta 1 --> 3)-D-GalNAc), SBA (Glycine max binding specificity alpha/beta-D-GalNAc > D-Gal), WGA (Triticum vulgare binding specificity (alpha-D-GlcNAc)n and sialic acid), ConA (Canavalia ensiformis binding specificity alpha-D-Man > alpha-D-Glc), LTA (Lotus tetragonolobus binding specificity alpha-L-fucose) and UEA 1 (Ulex europaeus binding specificity alpha-L-fucose). RESULTS: No significant differences were found in the glycoconjugates sugar residue content and distribution between the endometria of women < 30 years of age and those of women > 40. CONCLUSIONS: Our results suggest that human endometrium does not age, at least while cyclic hormonal stimulation and menstruation are present.

Histochemical characterization of the mucins of the epithelial cells in the chick embryo proventriculus.

The histochemical characteristics of the epithelial mucins, which are produced by the lining epithelium, the superficial glands and the compound glands of the proventriculus were investigated in the chick embryo from the 7th day of incubation to hatching. Results showed that the time of appearance of the carboxylated, sulphated and neutral mucosubstances differed at the three investigated sites. From the 13th to the 19th day of incubation, intense production of the various mucins was detected at the three sites. However, the modality of intracytoplasmatic storage and the production of mucins in both the cells of the lining epithelium and the superficial glands appeared to differ from those observed in the cells of the compound glands. At the 20th and 21st day, the mucins were primarily produced by the lining epithelium and the superficial glands, whereas production diminished in the compound glands. Furthermore, from the 7th to the 17th day of incubation, the cells of the principal ducts of the compound glands showed the same histochemical characteristics and modality of production as those of the glandular cells of the compound glands. On the other hand, from the 18th day to hatching, these cells assumed the same characteristics as those of the lining epithelium and of the superficial glands.

Identification of sugar residues in secretory glycoconjugates of the lining and glandular epithelium of the chick embryo gizzard using lectin histochemistry.

Using seven horseradish peroxidase-conjugated lectins we studied the distribution of glycoconjugate sugar residues in the lining and glandular epithelium of the gizzard in chick embryos from day 7 to 21 of incubation. At the outer layer of the lining epithelium all investigated sugar residues were present from day 7 onwards, except alpha-L-fucose, which was detected later. From day 9 some cells, which contained granules characterised by D-galactose-(beta 1--->3)-N-acetyl-D-galactosamine and N-acetyl-D-glucosamine, were observed in the inner part of the lining epithelium. Afterwards, although in different time periods, all investigated oligosaccharides were detected in intercellular spaces filled with mucous material. At first, the cells of the anlage of the tubular glands appeared to be characterised by D-glucosamine and alpha-D-mannose only. Afterwards, all the investigated sugar residues were detected. At hatching, the luminal glandular secretion showed all investigated sugar residues, except alpha-L-fucose, which was typical for the early formation of the tubular glands. Our data suggest that during the development of the gizzard three time periods of mucous release can be distinguished.

Magnetic resonance in the study of cranial nerves.

The course of cranial nerves was studied by means of magnetic resonance (MR), which allowed a multiplane visualization of the investigated structures. The obtained results showed that MR was an excellent method for visualization of the optic, trigeminal, facial and acoustica nerves. The oculomotor and the abducent nerves were detectable only in some regions where the contrast with the surrounding structures was greater. The glossopharingeus, vegus and accessory nerves were identifiable only in the first tract of emergency from the encephalic trunk and they could no be distinguished separately. The trochlear nerve was seldom visible whereas the olfactory nerve failed to be revealed. Further technological progress will allow for additional advances as regards the acquisition of knowledge concerning these important nerves.

The nasal respiratory epithelium in the human fetus: lectin histochemistry.

Lectin binding histochemistry was used to investigate the glycoconjugate saccharidic moieties in the respiratory epithelium of human fetuses ranging in age from the 8th to the 12th week of gestation. The distribution and changes in the sugar residues of the glycoconjugates at the level of the basal cells, the bathyprismatic cells and the bathyprismatic ciliated cells are reported. PNA and SBA allowed the identification of some 'elongated cells', which were characterized by lectin-reactive supranuclear granules. These epithelial cells, which have not previously been identified using conventional histochemical methods, might be the precursors of secretory cells.

Chick embryo metanephros: the glycosylation pattern as revealed with lectin conjugates.

Fragments of metanephros were taken from chick embryos and studied from the 7th to the 21st d of incubation. A battery of 7 different horseradish peroxidase-labelled lectins (PNA, ConA, DBA, SBA, LTA, WGA and UEA I) was used to analyse the distribution and changes of carbohydrate moieties in glycoconjugates along the metanephric nephron from the S-shaped body stage onwards. DBA and SBA reacted for a short time at some sites during the considered period of incubation. Provided WGA and ConA reacted with most of the nephric components, the almost ubiquitous presence of N-acetyl-D-glucosamine and alpha-D-mannose was demonstrated. SBA and LTA were found to be good markers of the proximal tubule. On the 12th d of incubation, sialic acid was found in the podocytes, capillary walls, and the connecting and collecting ducts. On the same day, other oligosaccharides were present at the nephric tubular components. The simultaneous presence of sialic acid and other sugar residues may be significant for the identification of the exact stage of incubation at which metanephric activity starts. The possible role of some sugar residues for the regulation of metanephric activity is discussed.

Light microscopic histochemical detection of sugar residues of glycoconjugates in human nasal mucosa using HRP-lectins.

A battery of horseradish peroxidase-conjugated lectins (ConA, WGA, PNA, SBA, DBA, UEAI and LTA) was used to study the content and distribution of glycoconjugate sugar residues of the normal human nasal mucosa. Our findings indicate that differences exist between the types of sugar residues found in the secretory product of the mucous goblet-like cells and those found in the mucous glands secretion. The saccharide content of the epithelial basal cells, which has never been previously investigated, is reported. Differences in the content and distribution of glycoconjugate sugar residues were found between human nasal mucosa and that of the other human extrapulmonary tracts.

Development of the chick tongue. A scanning electron microscopical investigation.

With the aim of clarifying certain morphological aspects of lingual development, chick tongues between the 8th day of incubation and hatching and also during the early postincubation period were investigated by means of the Scanning Electron Microscope. During this time, the tongue anlage underwent some remarkable morphogenetic changes, mostly involving the superficial epithelium but also including the appearance of the lingual glands at the level of the lingual root. With regards to the epithelium, it was possible to observe that in the first days of the incubation period examined, the superficial cells appeared dome-shaped, with microvilli on the apical surface; later they tended to become more flattened, and the microvilli were replaced by a thick net of microplicae. During the final days of incubation, and after hatching desquamative phenomena became evident. At no site of the tongue rudiment were taste buds ever observed, possibly because of the different functional role played by the avian tongue in comparison with that of the mammals.

The development of the caeca in the chick embryo. Morphometric and morphological remarks.

Quantitative aspects of the organogenesis of the chick embryo caecum (proximal and distal tracts) were investigated from the 8th to the 15th day of incubation. Histological sections were studied by means of a computerized morphometric system and a semiautomatic digital system (Videoplan 2). The total area of sections, of the lumen, the caecal wall and its components (subserous stratum, muscle layer, lamina propria, epithelium) and the thickness of the epithelium and muscle layer were measured. The mean +/- S.E.M. of the obtained values was calculated. The percentage of shrinkage, due to histological procedures, was calculated for each day considered. The mean values for each day of incubation were modified on the basis of the shrinkage percentage. The differences between the mean values of the areas of the proximal and distal tract were statistically evaluated. Exponential curves and the r coefficient were determined in order to evaluate the general growing pattern of the mean area of the caecal wall components as a function of age. The obtained results demonstrate that there are some differences between the proximal and distal tract of the caecal anlage in the growth pattern of the wall components. In the proximal caecal tract, the lamina propria showed the highest growth rate, while in the distal tract, the caecal wall component the subserous stratum did. Moreover, the caecal anlage showed different developmental behaviour from the ileum, duodenum and colon in the same chick embryo.

Histochemical detection of sugar residues in chick embryo developing lingual glands with horseradish-peroxidase conjugated lectins.

Tongue anlage were taken in chick embryos from the 7th to the 21st d of incubation and in 3 d old chicken. A battery of 7 different horseradish peroxidase-conjugated lectins (PNA, ConA, DBA, SBA, LTA, WGA, UEA I) was used to study the carbohydrate residues of glycoconjugates at the epithelial cells of the anterior and posterior lingual glands. Some sugar residues, detected at the surface of the epithelial cells in early developmental stages of glandular primordia, seemed to play a role in inducing and regulating the first differentiative steps of the glands. Differences in type, amount, time of appearance and cellular localization between the 2 groups of glands were detected. The group of the anterior lingual glands, adjacent to the entoglossal cartilage (paraentoglossal glands), showed some peculiar histochemical characteristics.

[Glycoconjugates of the cecal intestinal epithelium of the chick before and after hatching: histochemical study performed with peroxidase-conjugated lectins]. / I glicoconiugati nell'epitelio dell'intestino cieco del pollo prima e dopo la schiusa: studio istochimico eseguito con lectine coniugate a perossidasi.

A battery of seven different horseradish peroxidase-labelled lectins (PNA, ConA, DBA, SBA, LTA, WGA and UEA I) was used to study the distribution and changes of carbohydrate moieties of glycoconjugates in the caecal epithelium (proximal and distal tracts) of the chick embryo and of the 3 days old chicken. The chief results showed that: 1. The appearance of some sugar residues was earlier observed at the epithelium of the distal tract than the proximal one (Tab. 2). 2. The presence of sialic acid was detected only after hatching (Fig. 4, Tab. 2). 3. During the embryonic caecal development enterocytes and goblet cells were characterized by the presence of the same sugar residues (Tab. 2). 4. By a quantitative point of view, differences in sugar residues content between the epithelium of the proximal and distal tract were observed. The epithelial cells of the distal tract were generally characterized by an higher content of saccharide moieties (Tab. 2). 5. At the end of the incubation period and after hatching enterocytes and goblet cells showed differences in content of some sugar residues (Fig. 1-3, Fig. 5-8, Tab. 2).

Histochemical detection of sugar residues in the chick embryo mesonephros with lectin-horseradish peroxidase conjugates.

Fragments of mesonephros were taken from chick embryos and studied from the 4th to the 21st day of incubation. A battery of seven different horseradish peroxidase-labelled lectins was used to study the distribution of carbohydrate residues in glycoconjugates along the mesonephric nephron during the period of excretory activity and the period of involution. ConA and WGA reacted at every site of the nephron thus showing the ubiquitous presence of alpha-D-mannose and N-acetyl-D-glucosamine. SBA was a good marker of the proximal tubule. Other lectins, such as PNA and LTA, reacted only for a short time at some sites during the considered period of incubation. The presence of sialic acid was detected in the podocytes, capillary wall and mesangial cells. From the 10th-11th day of incubation changes were noted in the proximal tubule as shown by PNA reactivity. This may be significant as regards the exact stage of incubation during which the involution of mesonephros begins.

On the development of the chick embryo colon: a computerized morphometric study.

The quantitative aspects of the chick embryo colon organogenesis (proximal and distal tracts) have been studied from the 7th to the 15th day of incubation on histological sections by means of a computerized morphometric system. A semiautomatic digital system (Videoplan 2) was used. The area of total section, of the lumen, of the wall and its components (subserous stratum, muscle layer, lamina propria, epithelium) and the thickness of the epithelium and muscle layer were measured; the mean +/- S.E.M. of the obtained values was calculated. The percentage of shrinkage, due to histological procedures, was calculated for each day. The mean values for each day of incubation were modified on the basis of the shrinkage percentage. The differences between the mean values of the areas of the proximal and distal tract were statistically evaluated. Exponential curves and r coefficient were determined to evaluate the general growing pattern of the mean area of colonic wall components as a function of age. The main results demonstrate that in both tracts the component with the greatest growth was the muscle layer, while there are some differences between the proximal and distal tracts of the colonic anlage in the growing pattern of other wall components. The colonic anlage also shows a different developmental behaviour compared with that of the chick embryo ileum and duodenum.

Computerized morphometric analysis of the chick embryo duodenum organogenesis.

A computerized morphometric analysis was performed on histological sections of the proximal and distal limbs of the chick embryo duodenum, from the 8th to the 15th day of incubation, with the aim of evaluating the quantitative aspects of organogenesis. A semiautomatic digital system was used. The areas of total section, of the lumen, of the wall and of its components (subserous stratum, muscle layer, lamina propria, epithelium) and the thickness of the epithelium and muscle layer were measured; the mean +/- S.E.M. of the values obtained was calculated. The percentage of shrinkage, due to the histological procedures, was calculated for each day. The values obtained were added to the measurements of each area and thickness thus giving the true value of each parameter measured. The differences between the mean values of the areas of the proximal and the distal tract were statistically evaluated. Exponential curves and r coefficient were determined to evaluate the general growing pattern of the mean area of the components of the duodenal wall, as a function of age. The main results are the following: the areas of the wall components of the proximal limb are always greater than the corresponding areas of the distal limb of the duodenum. There are some differences between the proximal and the distal limbs of the duodenum in the growing pattern of the intestinal wall components. The anlage of the duodenum also has a different developmental pattern compared with that of the chick embryo ileum.