Search

Sialic acids (Sias) are a large and heterogeneous family of electronegatively charged nine-carbon monosaccharides containing a carboxylic acid and are mostly found as terminal residues in glycans of glycoproteins and glycolipids such as gangliosides. They are linked to galactose or N-acetylgalactosamine via α2,3 or α2,6 linkage, or to other Sias via α2,8 or more rarely α2,9 linkage, resulting in mono, oligo and polymeric forms. Given their characteristics, Sias play a crucial role in a multitude of human tissue biological processes in physiological and pathological conditions, ranging from development and growth to adult life until aging. Here, we review the sialylation status in human adult life focusing on the nervous and skeletal muscle tissues, which both display significant structural and functional changes during aging, strongly impacting on the whole human body and, therefore, on the quality of life. In particular, this review highlights the fundamental roles played by different types of glycoconjugates Sias in several cellular biological processes in the nervous and skeletal muscle tissues during adult life, also discussing how changes in Sia content during aging may contribute to the physiological decline of physical and nervous functions and to the development of age-related degenerative pathologies. Based on our current knowledge, further in-depth investigations could help to develop novel prophylactic strategies and therapeutic approaches that, by maintaining and/or restoring the correct sialylation status in the nervous and skeletal muscle tissues, could contribute to aging slowing and the prevention of age-related pathologies.

The vomeronasal system (VNS) is an accessory olfactory structure present in most mammals adhibited to the detection of specific chemosignals implied in social and reproductive behavior. The VNS comprises the vomeronasal organ (VNO), vomeronasal nerve and accessory olfactory bulb. VNO is characterized by a neuroepithelium constituted by bipolar neurons and supporting and stem/progenitor cells. In humans, VNO is present during fetal life and is supposed to possess chemoreceptor activity and participate in gonadotropin-releasing hormone neuronal precursor migration toward the hypothalamus. Instead, the existence and functions of VNO in postnatal life is debated. Vascular endothelial growth factor (VEGF) and its receptors (VEGFRs) have been demonstrated to play fundamental roles in various neurogenic events. However, there are no data regarding the localization and possible function of VEGF/VEGFRs in human fetal VNO. Therefore, this study was conceived to investigate the expression of VEGF/VEGFRs in human VNO in an early developmental period (9-12 weeks of gestation), when this organ appears well structured. Coronal sections of maxillofacial specimens were subjected to peroxidase-based immunohistochemistry for VEGF, VEGFR-1 and VEGFR-2. Double immunofluorescence for VEGF, VEGFR-1 or VEGFR-2 and the neuronal marker protein gene product 9.5 (PGP 9.5) was also performed. VEGF expression was evident in the entire VNO epithelium, with particularly strong reactivity in the middle layer. Strongly VEGF-immunostained cells with aspect similar to bipolar neurons and/or their presumable precursors were detected in the middle and basal layers. Cells detaching from the basal epithelial layer and detached cell groups in the surrounding lamina propria showed moderate/strong VEGF expression. The strongest VEGFR-1 and VEGFR-2 expression was detected in the apical epithelial layer. Cells with aspect similar to bipolar neurons and/or their presumable precursors located in the middle and basal layers and the detaching/detached cells displayed a VEGFR-1 and VEGFR-2 reactivity similar to that of VEGF. The basal epithelial layer exhibited stronger staining for VEGFRs than for VEGF. Cells with morphology and VEGF/VEGFR expression similar to those of the detaching/detached cells were also detected in the middle and basal VNO epithelial layers. Double immunofluorescence using anti-PGP 9.5 antibodies demonstrated that most of the VEGF/VEGFR-immunoreactive cells were neuronal cells. Collectively, our findings suggest that during early fetal development the VEGF/VEGFR system might be involved in the presumptive VNO chemoreceptor activity and neuronal precursor migration.

Several investigations have demonstrated neuroprotective effects of quercetin, a polyphenol widely present in nature, against neurotoxic chemicals, as well as in neuronal injury/neurodegenerative disease models. Most of these studies have been performed with quercetin aglycone and its metabolites, while scanty data are available on its glycosides. This study is aimed at investigating the neuroprotective effects of quercetin 3,4’-O-β- d -diglucoside (Q3,4’dG), isolated from the bulbs of the white cultivar (Allium cepa L.), using an in vitro model of human striatal precursor cells (HSPs), a primary culture isolated from the striatal primordium and previously characterized. To study the effect of Q3,4’dG on cell survival, HSPs were exposed to nutrient deprivation created by replacing culture medium with phosphate buffer saline (PBS). Our findings showed that Q3,4’dG treatment significantly promoted cell survival and strongly decreased apoptosis induced by nutrient deprivation, as evaluated by cell proliferation/death analyses. In addition, since the adhesive capacities of cells are essential for cell survival, the expression of some adhesion molecules, such as pancadherin and focal adhesion kinase, was evaluated. Interestingly, PBS exposure significantly decreased the expression of both molecules, while in the presence of Q3,4’dG this effect was prevented. This study provides evidence of a neuroprotective role exerted by Q3,4’dG and suggests its possible implication in sustaining neuronal survival for prevention and treatment of neurodegenerative disorders.

The vascular endothelial growth factor (VEGF)/VEGF receptor (VEGFR) system plays an important role in angiogenesis and osteogenesis during both skeletal development and postnatal bone growth and repair. Indeed, protein expression changes of this system could contribute to craniofacial defects commonly associated with a variety of congenital syndromes. Similarly to other craniofacial bones, mandible arises from neural crest cells of the neuroectodermal germ layer, and undergoes membranous ossification. Here, we report a case of left mandibular hypoplasia in a 42-year-old man treated with autologous bone graft from mandibular symphysis. After 3 months from surgical reconstruction, the protein expression of VEGF and receptors (VEGFR-1, -2 and -3) in regenerated bone tissue was evaluated by immunohistochemistry. At variance with the mandibular symphysis bone harvested for graft surgery, we observed de novo expression of VEGF and VEGFRs in osteoblasts and osteocytes from post-graft regenerating mandible bone tissue. In particular, while VEGFR-1 and VEGFR-3 immunopositivity was widespread in osteoblasts, that of VEGFR-2 was scattered. Among the three receptors, VEGFR-3 was the more intensively expressed both in osteoblasts and osteocytes. These findings suggest that VEGFR-2 might be produced during the early period of regeneration, while VEGFR-1 might participate in bone cell maintenance during the middle or late consolidation period. VEGFR-3 might, instead, represent a specific signal for ectomesenchymal lineage differentiation during bone regeneration. Modulation of VEGF/VEGFR signaling could contribute to graft integration and new bone formation during mandibular regeneration.

In this study the characterization of various types of sugar residues in normal, keratoconus and cross-linked human corneas was performed using immunohistochemical localization with lectins. Corneal samples were collected and divided into three groups: (1) normal corneas from cadavers; (2) keratoconic corneal buttons; (3) keratoconic corneal buttons treated with cross-linking. A series of lectins including: DBA, SBA, PNA, ConA, WGA, UEA I, GNA, DSA, MAA, SNA, were used in combination with chemical and enzymatic treatments. Compared with the normal corneas, N-acetyl-D-glucosamine increased in the keratoconus corneas. L-fucose increased and/or appeared in the keratoconus and the cross-linked corneas. N-acetyl-D-galactosamine was more abundant in the epithelium of keratoconus corneas, but was lacking in the keratoconus and cross-linked endothelium. D-galactose-(β1-4)-N-acetyl-D-glucosamine was absent in the whole stroma of the keratoconus corneas and in the deep layers of the cross-linked ones. Sialic acids increased in the keratoconus corneas and decreased in the cross-linked ones. These results showed altered glycosylation in the keratoconic corneas and partially similar glycosylation in the cross-linked corneas, compared to the normal ones. This suggests a role played by sugar residues in maintaining the corneal structure. The changes could be related to structural alterations in keratoconus. The present findings contribute to our understanding of the effect of cross-linking treatment of keratoconic corneas in therapeutic attempts to re-establish the normal corneal structure.

Objective Content and distribution of the oligosaccharides in the umbilical cord from pregnancies with altered glycemia were investigated. Study design A prospective cohort study was conducted in the Florence Policlinic of Careggi, Italy. Samples of cord from physiological pregnancies (n = 20), from pregnancies with minor degree of glucose intolerance (n = 20) and from pregnancies with gestational diabetes mellitus (GDM) treated with insulin (n = 20) were collected. Eleven lectins were used (ConA, WGA, PNA, SBA, DBA, LTA, UEA I, OOA, GSL II, MAL II and SNA) in combination with chemical and enzymatic treatments. Results Increase of N-acetyl- d -glucosamine and a loss of sialic acid in the umbilical cord of the cases with minor degree of glucose intolerance with respect to the other study groups was observed. d -Galactose(β1 → 3)-N-acetyl- d -galactosamine, N-acetyl- d -galactosamine and l -fucose were in less amount in both the pathological groups with respect to the control one. Conclusion The increase of some glycoconjugates carbohydrates and the loss of others in the umbilical cord from pregnancies with minor degree of glucose intolerance might be related to its morphofunctional alterations in a not diabetic altered glycemia. Moreover, the treatment with insulin in the GDM might play a role in restoring partially the normal glycosilation in the cord components in the attempt to renew some their functions.

Vascular endothelial growth factor (VEGF) is a well-known mediator that signals through pathways in angiogenesis and osteogenesis. Angiogenesis and bone formation are coupled during either skeletal development or bone remodeling and repair occurring in postnatal life. In this study, we examined for the first time the expression of VEGF in human fetal mandibular and femoral bone in comparison with the respective adult tissues. Similarly to other craniofacial bones, but at variance with the axial and appendicular skeleton, during development mandible does not arise from mesoderm but neural crest cells of the neuroectoderm germ layer, and undergoes intramembranous instead of endochondral ossification. By quantitative real-time PCR technique, we could show that VEGF gene expression levels were significantly higher in fetal than in adult samples, especially in femoral tissue. Western blotting analysis confirmed higher protein expression of VEGF in the fetal femur respect to the mandible. Moreover, immunohistochemistry revealed that in both fetal tissues VEGF expression was mainly localized in pre- and osteoblasts. Differential expression of VEGF in femoral and mandibular bone tissues could be related to their different structure, function and development during organogenesis.

Investigations mostly in animal models have shown a role of sialic acid in the morphology and functionality of skeletal muscle during development and adult life. Several studies in humans have been performed regarding changes in sialic acid expression in a particular pathology, hereditary inclusion body myopathy, leading to muscular weakness and atrophy, with a similar phenomenon appearing also in sarcopenia of aging. In this study the expression of monomeric and polymeric sialic acids was evaluated in human skeletal muscle during adult life. Surgical biopsies of the Quadriceps femoris muscle from men aged 18-25 years (young group; n=8) and men aged 72-78 (elderly group; n=10) were collected for analysis. Expression of sialic acids was evaluated using lectin histochemistry, associated with enzymatic and chemical treatments to characterize monomeric and polymeric sialic acids. The polysialic acid expression was also evaluated by immunohistochemistry. Various types of sialic acid in the muscle tissue, in different amounts in the study groups, were detected. Monomeric sialic acids decreased in the elderly group compared with the young group, whereas polysialic acid increased. Sialic acid acetylation was present only in the young group. These findings demonstrated that changes in the expression of sialic acids in skeletal muscle tissue may be related to morphofunctional modifications occurring during aging.

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.

The aim of the study was to investigate the content and distribution of sugar residues in placentas from pregnancies complicated by hypertensive disorders. Placentas from women with uncomplicated pregnancies (group 1), pregnancies complicated by gestational hypertension (group 2), pregnancies complicated by pre-eclampsia (group 3), pregnancies complicated by pre-eclampsia with HELLP syndrome (hemolysis, elevated liver enzymes and low platelets) (group 4) were collected. Lectins: ConA, WGA, PNA, SBA, DBA, UEA I, GNA, DSA, MAA, SNA, in combination with chemical and enzymatic treatments, were used. Data showed a decrease and/or lack of α-d-mannose, α-d-glucose and d-galactose-(β1-4)-N-acetyl-d-glucosamine in placentas from pre-eclampsia and pre-eclampsia with HELLP syndrome compared with control and hypertension cases. N-acetyl-d-galactosamine appeared and/or increased in placentas from hypertensive disorders. A different distribution of various types of sialic acid was observed in placentas from hypertensive disorders compared with the controls. In particular, placentas from pre-eclampsia, with and without HELLP syndrome, lacked the acetylated sialic acid side-chain. These findings demonstrate various alterations of the carbohydrate metabolism in the placentas from pregnancies complicated by different types of hypertensive disorders. This indicates correlation with the placental morpho-functional changes characteristic of these complications and with the degree of clinical severity.

Summary 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, Con A, DBA, SB A, 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.

A battery of six different horseradish peroxidase-labelled lectins (SBA, DBA, PNA, WGA, ConA and LTA) was used to study the distribution of carbohydrate residues in glycoconjugates during the development of the chick embryo tongue anlage (7th-21st day of incubation) and in the tongue of 3 day-old chicken, at the dorsal and ventral epithelium. From the 7th to the 16th day β-D-galactose, D-galactose-(β1 3)-N-acetyl-D-galactosamine, D-mannose and β-N-acetyl-D-glucosamine show the same epithelial distribution at two lingual aspects. From the 17th day to hatching the epithelium of the dorsal and ventral surfaces is characterized by a different distribution of sugar residues. The functional adaptation of the epithelium, as far as the saccharidic moletie distribution is concerned, seems to be completed at the 21st day of incubation.

The aim of this study was to investigate the distribution of the oligosaccharides of the glycoconjugates in placentas from pregnancies complicated by different degree of altered glycaemia. Placentas from women with physiological pregnancies (group 1), with pregnancies complicated by minor degree of glucose intolerance (group 2) and with pregnancies complicated by gestational diabetes mellitus (GDM) treated with insulin (group 3) were collected. Ten lectins were used (ConA, WGA, PNA, SBA, DBA, LTA, UEA I, GSL II, MAL II and SNA) in combination with chemical and enzymatic treatments. The data showed a decrease of sialic acid linked alpha(2-6) to galactose/N-acetyl-D-galactosamine and an increase of N-acetyl-D-glucosamine in the placentas of the pathological groups, in particular the group 3, comparing to the group 1. A decrease of L-fucose (LTA) and D-galactose-(beta1-3)-N-acetyl-D-galactosamine, and an increase and/or appearance of L-fucose (UEA I) and N-acetyl-D-galactosamine were observed in both the pathological groups, particularly in the group 2, with respect to the group 1. In GDM, and even in pregnancies with a simple alteration of maternal glycaemia, the changes in the distribution of oligosaccharides could be related to alteration of the structure and functionality of the placenta.

The aim of the present study was to determine the expression of vascular endothelial growth factor (VEGF) family receptors (VEGFR) in placentas from pregnancies complicated by hypertensive disorders of different clinical severity. Placental tissue from women with gestational hypertension, pre-eclampsia, pre-eclampsia with haemolysis, elevated liver enzymes and low platelets (HELLP syndrome) and normotensive women, as a control group, was examined. Immunohistochemical techniques, reverse transcription–polymerase chain reaction and western blot were used to evaluate receptor expression. In cases with gestational hypertension, as well as in control cases, VEGFR-1 and VEGFR-3 immunoreactivity was detected in all placental components, whereas in placentas from the pre-eclampsia and pre-eclampsia with HELLP syndrome groups, VEGFR-1 and VEGFR-3 immunoreactivity was detected only in some portions of trophoblast and/or some vessels and/or clusters of stromal cells. In the control group, VEGFR-2 immunoreactivity was observed only in the vessels, whereas the hypertensive groups showed VEGF-2 immunoreactivity also in trophoblast and stromal cells. The mRNA levels of the three receptors in the group with gestational hypertension were higher with respect to those in the control group. Placentas from pregnancies with pre-eclampsia showed lowest mRNA expression levels, whereas placentas from women with pre-eclampsia plus HELLP syndrome showed higher mRNA expression levels with respect to the three other groups. Receptor protein levels were lower in pathological cases compared with levles in the control group. These findings demonstrate a dysregulation of placental expression of VEGF family receptors related to the degree of clinical severity of the hypertensive disorder.

The oligosaccharide distribution of the glycoconjugates was investigated in placental tissue of pregnancies complicated by intrauterine growth retardation (IUGR) with absent or reversed flow in the umbilical artery (ARED), between the 29 and the 37 weeks of pregnancy. Placentae of a gestational age-matched group of normally grown pregnancies was also selected as control group. For this purpose a battery of seven HRP-conjugated lectins was used (DBA, SBA, PNA, ConA, WGA, LTA and UEA I). Our data showed that alpha-D-mannose (ConA), N-acetyl-D-glucosamine (WGA), beta-N-acetyl-D-galactosamine (SBA), alpha-L-fucose (LTA and UEA I) were present in less amount or were not present in the trophoblast and/or in the endothelial cells of the pathological group compared to the control one. The trophoblast basement membrane and/or basal plasma membrane of the pathological placentae were characterized by the presence of alpha-D-mannose (ConA), N-acetyl-D-glucosamine (WGA), sialic acid and D-galactose-(beta1--3)-N-acetyl-D-galactosamine (neuraminidase-PNA), only in some tracts, in all the weeks of gestation. In the control placentae these sugar residues were present in the whole basement membrane and/or basal plasma membrane from 31 or 35 weeks. The Hofbauer cells of the pathological placental tissue showed a less amount or lack of alpha-D-mannose (ConA), beta-N-acetyl-D-galactosamine (SBA) and alpha-L-fucose (UEA I) compared to the control ones. These results suggest that a less amount or lack of some sugar residues may contribute to restricted placenta growth and development and thus reduced efficiency in maternal-fetal exchanges of gases and metabolites.

The content, distribution and changes of the glycoconjugates sugar residues in the ovaries of chick embryos, from the 8th day of incubation to hatching and in 1-day old chick, were investigated. For this purpose, a battery of seven HRP-conjugated lectins was used (DBA, SBA, PNA, ConA, WGA, LTA and UEA I). Our data showed that SBA was a marker of the most immature oogonia in the ovarian cortex and medulla. The reactivity with ConA appeared to characterize the cells immediately prior to as well as during the meiotic division, as demonstrated by the presence of alpha-D-mannose at the "Balbiani bodies" in the oogonia of the ovarian cortex. The detection of Con A and SBA reactivity corresponded to maturative stages of the early oogonia in different cortical zones of the chick ovary. Our data also revealed that PNA seemed to be a marker of the degenerating oogonia located in the ovarian medulla. Moreover, PNA binding was a characteristic finding in the endothelial cells of the vessels located in the compact portion of the medulla in the left ovary, from the 8th to the 21st day of incubation and after hatching; PNA reactivity was only seen from the 16th day onwards in the endothelial cells of the cortex. During the whole considered period of incubation and after hatching, reactivity with UEAI, LTA and DBA was never detected.

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.

A battery of horseradish peroxidase-conjugated lectins (PNA, SBA, DBA, WGA, ConA, LTA and UEAI) was used to study the distribution of glycoconjugate sugar residues in the exocrine and endocrine pancreas of chick embryos, 1-day-old chicks and adult animals. During the period of incubation considered here, the time of appearance and changes in the oligosaccharides were noted in the acinar cells, in the endocrine cells of the "light islets" and in the capillary endothelial cells. In the developing exocrine pancreas, the basolateral surface of the acinar cells reacted with PNA, WGA, LTA and ConA for the entire period of incubation while SBA, WGA, LTA and ConA reactivity was detected in the apical plasma membrane. Zymogen granule membrane reacted with PNA, WGA and LTA from the 12th day onward, while ConA reactivity was detectable from the 7th day onward. Owing to its early appearance, alpha-D-mannose, revealed by ConA lectin, might in some way contribute to the maturation of the zymogen granule. In the 1-day-old chick and in the adult the cellular surface reacted with WGA and ConA. The zymogen granule membrane reacted with PNA, WGA and ConA in the 1-day-old chick. In the adult, WGA and ConA reactivity was observed in the zymogen granule membrane, whereas at this site PNA reactivity was revealed only after neuraminidase digestion. The hypothesis formulated by others that granule membrane lectin reactivity is equal to the apical membrane reactivity, owing to exocytotic processes, is not consistent with our results in the chick embryos, the 1-day-old chick and the adults. The surface and the granules of the islet beta cells in different periods of incubation reacted with WGA, SBA and, after neuraminidase treatment, with PNA. The same reactivity was seen also in 1-day-old chick while in the adult the granules of the beta cells also reacted with ConA. These findings show the achievement of an almost adult-like state of glycosylation of the glycoconjugates in the beta cells at an early stages of development.

Summary Lectin histochemistry was used to investigate the glycoconjugate saccharidic moieties in the skin of human fetuses ranging in age from the 8th to the 13th week of gestation. For this purpose, seven HRP-conjugated lectins were employed (Con A, WGA, PNA, DBA, SBA, UEAI and LTA). The distribution and changes of the sugar residues of glycoconjugates at the level of the various layers of the fetal epidermis, as well of the underlying mesenchyme were studied. We have shown that UEAI reactivity is a characteristic finding of the cells of the intermediate layer at the 11th and 12th week of gestation. LTA reactivity was detected only at the free border of the peridermal cells from the 10th to the 13th week of gestation. We demonstrated the presence of PNA binding sites at both the basal and peridermal cells from the 8th week of gestation, whereas other authors have stated that the appearance of PNA reactivity coincides with the onset of fetal skin stratification (11th–12th week of gestation). Moreover, some typical features in lectin reactivity at the level of the mesenchymal cells and fibres have been pointed out.

To evaluate if the peri-implantation endometrium shows age variations in lectin patterns, which suggest possible age variations in embryo-maternal recognition.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-GalNAcD-Gal), WGA (Triticum vulgare binding specificity (alpha-D-GlcNAc)n and sialic acid), ConA (Canavalia ensiformis binding specificity alpha-D-Manalpha-D-Glc), LTA (Lotus tetragonolobus binding specificity alpha-L-fucose) and UEA 1 (Ulex europaeus binding specificity alpha-L-fucose).No significant differences were found in the glycoconjugates sugar residue content and distribution between the endometria of women30 years of age and those of women40.Our results suggest that human endometrium does not age, at least while cyclic hormonal stimulation and menstruation are present.

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.

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.

The aim of the present study was to investigate the distribution of the glycoconjugates sialoderivatives in the human testis. Orchidectomy specimens from men aged 18–30 years (Group 1) and from men aged 70–93 years (Group 2) were obtained at autopsy. The study was performed using digoxigenin-labelled lectins, namely Maackia amurensis II lectin (MAA), Sambucus nigra agglutinin (SNA) and Arachis hypogaea lectin (PNA), in addition to enzymatic and chemical treatments (neuraminidase, KOH–neuraminidase, mild oxidation–neuraminidase, mild oxidation–KOH–neuraminidase, strong oxidation–neuraminidase, strong oxidation–KOH–neuraminidase), to characterise the different glycosidic linkages of the sialoderivatives and to obtain information regarding their structure. In all Group 2 samples, sialic acids linked α-2,3 to galactose and α-2,6 to galactose/N-acetyl-d-galactosamine (Gal/GalNAc), revealed by MAA and SNA, respectively, were observed in testicular interstitial tissue and in the lamina propria. Sialic acid linked α-2,6 to Gal/GalNAc was detected in only some samples from Group 1. After treatment, PNA showed structural changes and/or the gradual disappearance of sialic acid linked to d-galactose-β(1–3)-N-acetyl-d-galactosamine in testicular components with aging. These findings indicate that changes in the metabolism of sialoderivatives in the testis could be related to morphofunctional changes in various testicular components typical of this organ during aging. This suggests that sialoderivatives are important in the functionality of the mature testis in men, as well as its involution.

The aim of the present study was to determine the expression of vascular endothelial growth factor (VEGF) receptors VEGFR-1, VEGFR-2 and VEGFR-3 in placentas from pregnancies complicated by altered glycaemia. Placentas from women with physiological pregnancies (Group 1), pregnancies complicated by minor degree of glucose intolerance (MDGI, Group 2) and by gestational diabetes mellitus (GDM) treated with insulin (Group 3) were collected. Immunohistochemistry, RT–PCR and western blot were employed to evaluate receptor expression. In the three study groups, VEGFR-1 immunoreactivity was detected in all the placental components. VEGFR-2 immunoreactivity was observed in the vessels of all the placentas from Groups 1 and 2, but only in some placentas of Group 3. VEGFR-3 reactivity was observed in all the components of Group 1; in Groups 2 and 3 reactivity was observed in some portions of the trophoblast or the whole trophoblast, and in the stroma. VEGFR-1 and VEGFR-2 mRNA levels in Groups 2 and 3 were significantly higher compared with Group 1, whereas those of VEGFR-3 were significantly lower. Receptor protein levels were significantly lower in Groups 2 and 3 compared with Group 1. These findings demonstrated dysregulation of expression of the three placental receptors, both in GDM and in MDGI.