Acid Glycosaminoglycans Research Articles

Morphological features of enamel in fluorosis of different degrees of severity

The occurrence of dental fluorosis is facilitated by a violation of enamel mineralization caused by fluorides, which enter the human body in excess during its development and formation and have a toxic effect on enamel blasts. However, the molecular mechanisms involved in the pathogenesis of fluorosis are not fully understood. Enamel formation is a complex process involving cell proliferation and differentiation through epithelial-mesenchymal sequential secretion of matrix proteins, tissue-specific transport of ions including calcium and fluoride, and precipitation and alignment of enamel crystals through interactions between organic and inorganic molecules. Understanding the morphological features of enamel changes during fluoride intoxication of the human body in the endemic region allows us to clearly understand the need for a comprehensive solution to this medical and social problem. The aim was to study the morphological features of enamel in fluorosis in residents of the endemic region of Ukraine, in particular the Poltava region. The work examines different groups of teeth (both intact and affected by fluorosis) removed for orthodontic or clinical indications in men and women aged 17 to 40 years. Morphological signs were studied first on native, and later on histochemically stained sections. It was established that the violation of the structure of the enamel layer of the teeth in mild and severe fluorosis is characterized by both partial and complete violation of the movement of the enamel prisms with signs of destruction. Fragmentation and homogeneity throughout the entire thickness were found in some areas of the enamel. When evaluating histochemically stained sections of teeth affected by fluorosis, it was established that dystrophic changes in the enamel structure and accumulation of acidic glycosaminoglycans in the lesions are more characteristic of mild and moderate forms. Under the conditions of a severe form of fluorosis, complete destruction of the prisms, fragmentation of the lamella, homogenization of areas on the entire enamel layer, which is due to the uneven distribution of acidic mucopolysaccharides, have been established. Morphologically and histochemically dystrophic changes in the areas of the affected enamel are confirmed by uneven distribution and accumulation of acidic glycosaminoglycans. An assessment of the effect of fluoride intoxication on the state of tooth enamel was carried out, which will allow to expand the possibilities of preventive measures for related specialists, as well as to create and develop additional treatment methods that will contribute to the improvement of physical and aesthetic indicators of teeth. dental health.

Acidic Osteoid Templates the Plywood Structure of Bone Tissue.

Bone is created by osteoblasts that secrete osteoid after which an ordered texture emerges, followed by mineralization. Plywood geometries are a hallmark of many trabecular and cortical bones, yet the origin of this texturing in vivo has never been shown. Nevertheless, extensive in vitro work revealed how plywood textures of fibrils can emerge from acidic molecular cholesteric collagen mesophases. This study demonstrates in sheep, which is the preferred model for skeletal orthopaedic research, that the deeper non-fibrillar osteoid isorganized in a liquid-crystal cholesteric geometry. This basophilic domain, rich in acidic glycosaminoglycans, exhibits low pH which presumably fosters mesoscale collagen molecule ordering in vivo. The results suggest that the collagen fibril motif of twisted plywood matures slowly through self-assembly thermodynamically driven processes as proposed by the Bouligand theory of biological analogues of liquid crystals. Understanding the steps of collagen patterning in osteoid-maturation processes may shed new light on bone pathologies that emerge from collagen physico-chemical maturation imbalances.

High-Performance Polarization Microscopy Reveals Structural Remodeling in Rat Calcaneal Tendons Cultivated In Vitro.

Collagenous tissues exhibit anisotropic optical properties such as birefringence and linear dichroism (LD) as a result of their structurally oriented supraorganization from the nanometer level to the collagen bundle scale. Changes in macromolecular order and in aggregational states can be evaluated in tendon collagen bundles using polarization microscopy. Because there are no reports on the status of the macromolecular organization in tendon explants, the objective of this work was to evaluate the birefringence and LD characteristics of collagen bundles in rat calcaneal tendons cultivated in vitro on substrates that differ in their mechanical stiffness (plastic vs. glass) while accompanying the expected occurrence of cell migration from these structures. Tendon explants from adult male Wistar rats were cultivated for 8 and 12 days on borosilicate glass coverslips (n = 3) and on nonpyrogenic polystyrene plastic dishes (n = 4) and were compared with tendons not cultivated in vitro (n = 3). Birefringence was investigated in unstained tendon sections using high-performance polarization microscopy and image analysis. LD was studied under polarized light in tendon sections stained with the dichroic dyes Ponceau SS and toluidine blue at pH 4.0 to evaluate the orientation of proteins and acid glycosaminoglycans (GAG) macromolecules, respectively. Structural remodeling characterized by the reduction in the macromolecular orientation, aggregation and alignment of collagen bundles, based on decreased average gray values concerned with birefringence intensity, LD and morphological changes, was detected especially in the tendon explants cultivated on the plastic substrate. These changes may have facilitated cell migration from the lateral regions of the explants to the substrates, an event that was observed earlier and more intensely upon tissue cultivation on the plastic substrate. The axial alignment of the migrating cells relative to the explant, which occurred with increased cultivation times, may be due to the mechanosensitive nature of the tenocytes. Collagen fibers possibly played a role as a signal source to cells, a hypothesis that requires further investigation, including studies on the dynamics of cell membrane receptors and cytoskeletal organization, and collagen shearing electrical properties.

Pathomorphological characteristics of cotyledons with weak contrasting of the blood flow in the placenta of women who have undergone exacerbation of cytomegalovirus infection during the second trimester of pregnancy

Aim. To give pathomorphological characteristics of placental cotyledons with weakly contrasted bloodstream in women who had an exacerbation of cytomegalovirus infection (CMVI) in the second trimester of pregnancy.Materials and methods. A study was made of involutive-destructive processes in 153 cotyledons of the placenta with well and weakly contrasted bloodstream in women with pregnancy, uncomplicated and complicated by exacerbation of CMVI at 21-24 weeks of gestation. The first group included 36 cotyledon placentas from women with CMV-seronegative uncomplicated pregnancy with well contrasted blood vessels; the second group – 67 cotyledon placentas from patients with chronic compensated placental insufficiency (CCPI) and poorly contrasted vascular network; the third group – 30 cotyledon placentas from women with chronic subcompensated placental insufficiency (CSPI) with indistinctly visualized blood vessels; the fourth group – 20 cotyledon placentas from patients with chronic decompensated placental insufficiency (CDPI). Dosed introduction of red lead on drying oil (1:3) into the blood vessels of the placenta was carried out through the umbilical cord vein. Obtaining X-ray images of cotyledons with well and weakly contrasted blood vessels in the marginal part of the organ was carried out on the apparatus RUM-20M with X-ray image intensifier Sapphire (Russia). Cotyledon biopsy specimens for histometry and histochemical analysis were taken before their contrasting, and for survey microscopy – after injection of the contrast mass.Results. In the first group, the amount of fibrinoid in the stroma of the villi was 1.35±0.09%, perivillous fibrinoid − 1.02±0.08% in the absence of pseudonecrosis and calcifications. In the second group, unlike the first one, cotyledons prevailed, in which in 40-50% of cases weakly expressed contours of blood vessels were found, an increase in the specific volume of stromal fibrinoid by 1.82 times was observed (p<0.001), and fibrinoid around villi by 2.04 times (p<0.001); pseudonecrosis accounted for 2.29±0.13%, and calcificates − 1.50±0.12%. In the third group, in comparison with the second one, cotyledons were more common with 50-70% of indistinctly contoured vessels, villi with edematous changes and stromal collagenization; the concentration of stromal fibrinoid increased by 1.47 times (p<0.001), the proportion of perivillous fibrinoid – 1.46 times (p<0.001), pseudonecrosis – 1.41 times (p<0.001) and the accumulation of calcium salt – 1.57 times (p<0.001). In the fourth group, compared to the third one, cotyledons were more common, in which more than 70% of weakly contrasted blood vessels were determined, while the number of collagen fibers and acid glycosaminoglycans increased in the villi; there was an increase in the amount of fibrinoid around the villi by 1.24 times (p<0.01), pseudonecrosis – by 1.23 times (p<0.05) and calcificates – by 1.32 times (p<0.01).Conclusion. In women with an exacerbation of CMVI in the second trimester of gestation and CSPI, in contrast to patients with a similar infectious disease and CCPI, a decrease in the flow of contrast into the bloodstream of cotyledons is due to more pronounced edema, the formation of collagen fibers, fibrinoid and calcium salts in the stroma of the villi, as well as perivillous fibrinoid. In CDPI of cytomegalovirus etiology, compared with CSPI, weak contrasting of the vascular bed of cotyledons is associated with increasing changes in the stroma involved in the regulation of the resistance of draining veins and intracotyledon blood vessels.

Morphological and functional relationship between the orbital gland and olfaction in Upupa epops (hoopoe) and Bubulcus ibis (cattle egret).

This study used both anatomical and histological techniques to investigate the orbital gland's topographic relationship with the surrounding system, using the hoopoe and cattle egret as biological models. Hoopoe has a spindle-shaped lacrimal gland that is suspended on the lateral edge of the frontal bone, whereas cattle egret has a tiny lacrimal gland that is embedded posteriorly within the periorbital fascia. The hoopoe's lacrimal gland has a single duct that runs parallel to the nasolacrimal duct and opens into the posterior nostril hole. In the cattle egret, the tubule-alveolar secretory components comprise neutral and acid glycosaminoglycan. In addition, the Harderian gland is found in both these species, but their draining ducts differ; the Harderian gland of the hoopoe opens into the anterodorsal to the conjunctival fornix, whereas the Harderian gland of the egret opens anteriorly. In both hoopoe and egret, the secretions of Harderian gland include neutral and acid glycosaminoglycan. The Harderian gland is categorized as type II in hoopoe and type I in cattle egrets. The present results concluded that both orbital glands of two bird species studied play an essential role in eye health, where cleaning and lubrication of the cornea surface. Furthermore, the lacrimal gland's location and secretory features may strengthen the olfactory sensitivity of hoopoe, which relies heavily on scent to locate their food, whereas egret relies heavily on visual cues.

Anti‑chondrocyte apoptosis effect of genistein in treating inflammation‑induced osteoarthritis.

Osteoarthritis (OA) is a chronic disease that is mainly characterized by chondrocyte degeneration. Inflammatory mediators participate in the development of OA, leading to chondrocyte apoptosis and destruction of the cartilage. Genistein is the major active component of isoflavone, with a chemical composition and a biological effect that is similar to that of estrogens, which prevents the degradation of cartilage; however, its underlying mechanisms of action remain unknown. The aim of the present study was to investigate the anti-apoptotic effects of genistein on chondrocytes for the treatment of inflammation-induced OA. Interleukin (IL)-1β was used to establish a chondrocyte OA model. After treatment with different concentrations of genistein, western blotting identified that expression levels of collagen II and aggrecan were increased in a concentration-dependent manner, while caspase 3 expression gradually decreased after genistein application. Moreover, flow cytometry and ELISA results demonstrated that genistein could decrease chondrocyte apoptosis and reduce the levels of tumor necrosis factor (TNF)-α in a dose-dependent manner. Furthermore, the in vitro data were evaluated in an OA rat model. Genistein increased the collagen and acid glycosaminoglycan content, as well as decreased the levels of TNF-α and IL-1β. Genistein also promoted the expression levels of collagen II and aggrecan in the articular cartilage, and decreased the expression of caspase 3, thus alleviating cartilage degradation. In conclusion, the results indicated that genistein mediated inflammation and had an anti-apoptotic role in treating OA. Therefore, genistein may serve as an alternative treatment for OA.

Structural organization of the umbilical cord vascular connective tissue in cytomegalovirus infection in the third trimester of pregnancy

Aim. To study the structural organization of the connective tissue of the vein wall of the umbilical cord vessels from seropositive to cytomegalovirus (CMV) women with an exacerbation of infection in the third trimester of pregnancy. Materials and methods . There was done the study of the activity of glycosaminoglycans in the connective tissue of the vascular wall of the vein and the content of elastase in the umbilical cord homogenate obtained during childbirth at 38-39 weeks of pregnancy from 35 CMV-seropositive women with an exacerbation of infection in the third trimester of pregnancy (the main group), and from 32 CMV-seronegative women (the control group). The material for the histo-chemical study was the sections of the venous wall of the umbilical cord vessels adjacent to the placenta. The activity of glycosaminoglycans was studied by Steedman, the content of elastase was studied by enzyme immunoassay. Results. The study of umbilical cord tissue extracts revealed a 1.3-fold increase in immune enzyme elastase indices (p<0.001) in the group of CMV-seropositive women with an exacerbation of infection in the third trimester of pregnancy. Histochemical examination of the same umbilical cords revealed an increase in the intensity of the reaction to acidic glycosaminoglycans located in the subendothelial layer, in the intermuscular layers of connective tissue, in the cytoplasm of fibroblasts and in the smooth muscle cells of the inner layer of the umbilical vein muscles. The most pronounced reaction to glycosaminoglycans was detected in the collagen fibers of the subendothelial layer of the umbilical vein. In addition, areas of pronounced destruction and desquamation of endothelial cells were detected. The connective tissue of the subendothelial layer was characterized by the disappearance of the tortuosity of collagen fibers with signs of atrophy and a decrease in the number of cell elements in the vascular wall. Conclusion. Based on the results of the study, it was shown that the exacerbation of CMV infection in the third trimester of pregnancy is characterized by an increase in the enzymatic activity of elastase and the production of acid glycosaminoglycans in the connective tissue of Wharton’s jelly, which leads to the destruction of collagen fibers and disturbance of the framework and elasticity of the vascular wall of the umbilical vein.

Improvements in Skin Quality Biological Markers in Skin Explants Using Hyaluronic Acid Filler VYC-12L.

Background:Hyaluronic acid (HA), both crosslinked and uncrosslinked, is used clinically to treat fine lines and provides additional improvements in skin quality attributes. The purpose of this study was to assess potential early differences in the expression of biological markers of skin quality in living human skin explants injected with uncrosslinked and crosslinked HA gels.Methods:Living human skin explants injected with VYC-12L or noncrosslinked HA with mannitol (HYD) and noninjected controls were assessed via microscopy, histology, and immunohistochemistry on days 3 and/or 8 for biological markers of elasticity (collagen density, elastin, fibrillin-1) and hydration [aquaporin-3, acidic glycosaminoglycans (GAGs), HA]. Hydration was also assessed via a corneometer probe on days 0, 1, 2, and 8.Results:On day 3 versus controls, VYC-12L moderately increased collagen density in the upper reticular dermis and clearly increased fibrillin-1 expression, with slight increases persisting on day 8. Increases with HYD were smaller and did not persist on day 8. Both VYC-12L and HYD increased aquaporin-3 expression and GAG content on days 3 and 8, but VYC-12L produced greater GAG increases in the reticular dermis. Day 8 instrument-assessed hydration increased by 49% and 22% for VYC-12L and HYD, respectively. Elastin expression in oxytalan and elaunin fibers was unchanged. Upper-dermal HA reductions suggested HA injection-induced hyaluronidase expression.Conclusion:VYC-12L produced greater, more lasting improvements in biological markers of skin quality than HYD.

Correction of cicatricial changes in subacute stage of myocardial infarction with using of dispersed allogenic biomaterial

Aim. To identify the features of myocardial remodeling after implantation of a dispersed allogenic biomaterial (DAB) in the subacute stage of experimental infarction and determine the role of decay products in the formation of regenerate.Material and methods. Male rats in the control group (n=30) 5 days after coronary occlusion received physiological saline (intramyocardially), in the experimental group (n=30) — 3 mg of DAB. Myocardial tissues were examined after 7 14, 50 days from the beginning of the experiment using histological, immunohistochemical and statistical methods.Results. In the experimental group, the cicatrix area was reduced by 1,66 times. The products of DAB biodegradation were collagen, sulfated (dermatan- and keratan sulfate), as well as non-sulfated (hyaluronic acid) glycosaminoglycans. Used DAB served as a promoter of the early proliferative phase of inflammation and had an anti-inflammatory effect. DAB particles were a chemoattractant of stem and progenitor myocardial cells in the subacute stage of myocardial infarction. It contributed to their differentiation and integration into the tissue.Conclusion. The products of biodegradation of allogeneic biomaterial, administered intramyocardially in the subacute stage of infarction, contribute to the formation of muscular-connective tissue regenerate and effectively prevent its cicatricial devolution.

Collagen, glycosaminoglycans and matrix metalloproteinase-2 and metalloproteinase-9 in the cervix of the ewe during prepubertal development.

The tortuous nature of the ovine cervix restricts the transcervical passage of the cannula, and many studies have aimed to understand the endocrine mechanism of the remodelling of cervical tissue in adult ewe. However, little is known about the remodelling of the cervical tissue during the prepubertal development of the lambs. To obtain histochemical and biochemical evidence about the nature of the prepubertal development of the cervix of the ewe, cervices of Corriedale lambs obtained at 0, 1, 2, 4, 6 and 8months of age (n=5 to 6 in each) were processed. Neutral and acidic glycosaminoglycans (by PAS-Alcian stain) were weakly in the cervical stroma and not shown change during the development, whereas the percentage volume of fibrillar collagen (by van Gieson stain) increases throughout the experimental period in the superficial fold stroma and deep wall stroma (p<0.05). The relative cervical weight (g/kg of body weight) and the collagen concentration (by spectrophotometry, mg/mg wet tissue) showed an early decreasing phase from months 0 to 4 and a later increasing phase from months 4 to 8 (p<0.05). The latent form of matrix metalloproteinase-2 (MMP-2) detected by gelatin zymography (ng/mg protein) decreased from months 0 to 2 and increased from months 4 to 8, whereas the activated form decreased from months 0 to 2, remained low until month 6 and then recovered on month 8 (p<0.0001). Data suggest that the relative cervical weight biphasic pattern during the development is related to MMP-2-dependent changes in the collagen content.

Histochemical picture in the skin of rats 1, 3, 7, 14, 21 and 30 days after burning of ІІ-ІІІ degrees on the background of injection during first 7 days of 0.9% NaCl solution

For pathogenetically grounded therapy of burn disease, aimed at reducing the severity of the pathological process, the rapid recovery of microcirculation and stimulation of reparative processes, it is necessary to study the fine mechanisms of reparative regeneration in the skin on the microscopic and biochemical levels. The purpose of the study is to investigate the state of elastic and collagen fibers, as well as glycoproteins and glycosaminoglycans in the intercellular substance of the dermis of rats 1, 3, 7, 14, 21 and 30 days after the burning of the II-III degree, with the background of the injection during first 7 days of 0.9% NaCl solution. An experimental study of structural changes in the skin after burn injury was performed on laboratory white rats, males weighing 150-160 g. Rats were divided into 2 groups, which previously, under the conditions of propofol anesthesia 60 mg/kg internally, catheterization of the femoral vein and depilation lateral surfaces of the trunk of rats. Group 1 - animals without thermal trauma of the skin, which once a day for the first 7 days were injected intravenous infusion of 0.9% NaCl solution in a dose of 10 ml per kg. Group 2 - animals who were also injected once a day with the first 7 days of infusion of 0.9% NaCl solution in a dose of 10 ml per kg after 2-3 degree skin burns, with a total area of 21-23% of the body surface of the rat. Euthanasia of rats was performed after propofol anesthesia (60 mg/kg i/v) by decapitation. Changes in the histochemical structure of bits of skin from the edge of the wound were studied after 1, 3, 7, 14, 21, and 30 days from the beginning of the experiment. Histological sections 5-6 microns thick were stained with Weigert’s elastic stain and dyed with picric acid by Van Gieson’s method (for the detection of elastic and collagen fibers); the PAS + Hale reaction was performed by the Moury method (detection of glycoproteins and glycosaminoglycans in the intercellular substance of the dermis). The histochemical investigations of the area of the II-III degree of skin damaged by burns on the background of the injection of 0.9% NaCl solution found that the degree of change depends on the duration of the experiment. In the early stages of the experiment, the reorganization of the intercellular substance of the dermis manifests itself as damage to the fibrous structures of its papillary and reticular layers. Histochemically, the components of amorphous connective tissue substance are dominated by glycoproteins, the content of acidic glycosaminoglycans is small. In the late stages of the experiment (after 14, 21 and 30 days) in the connective tissue of the affected skin in the central and marginal regions poorly structured collagen and elastic fibers are observed. The pronounced “Hale”-positive coloration reflects an increase in the content of sulfated glycosaminoglycans in the amorphous substance of the connective tissue. Thus, the experimental thermal trauma of skin II-III degree on the background of the introduction during the first 7 days, 0.9% of the NaCl solution is histochemically characterized by reorganization of the intercellular substance of the connective tissue of the dermis. In the early stages of the experiment there is damage to the fibrous structures of the papillary and reticular layers, the prevalence of glycoproteins and the insignificant content of acidic glycosaminoglycans in the amorphous substance, indicating a significant inhibition of adaptive-compensatory processes. In the long term, after the thermal defeat in the dermis, histochemically, an increase in the amount of acid glycosaminoglycans is detected and the PAS positivity of the collagen fibers of the intercellular substance is well expressed. This indicates a slow updating of the amorphous substance and fibrous structures of the connective tissue of the dermis, a violation of regenerative processes.

CCL19 with CCL21-tail displays enhanced glycosaminoglycan binding with retained chemotactic potency in dendritic cells.

CCL19 is more potent than CCL21 in inducing chemotaxis of human dendritic cells (DC). This difference is attributed to 1) a stronger interaction of the basic C-terminal tail of CCL21 with acidic glycosaminoglycans (GAGs) in the environment and 2) an autoinhibitory function of this C-terminal tail. Moreover, different receptor docking modes and tissue expression patterns of CCL19 and CCL21 contribute to fine-tuned control of CCR7 signaling. Here, we investigate the effect of the tail of CCL21 on chemokine binding to GAGs and on CCR7 activation. We show that transfer of CCL21-tail to CCL19 (CCL19CCL21-tail ) markedly increases binding of CCL19 to human dendritic cell surfaces, without impairing CCL19-induced intracellular calcium release or DC chemotaxis, although it causes reduced CCR7 internalization. The more potent chemotaxis induced by CCL19 and CCL19CCL21-tail compared to CCL21 is not transferred to CCL21 by replacing its N-terminus with that of CCL19 (CCL21CCL19-N-term ). Measurements of cAMP production in CHO cells uncover that CCL21-tail transfer (CCL19CCL21-tail ) negatively affects CCL19 potency, whereas removal of CCL21-tail (CCL21tailless ) increases signaling compared to full-length CCL21, indicating that the tail negatively affects signaling via cAMP. Similar to chemokine-driven calcium mobilization and chemotaxis, the potency of CCL21 in cAMP is not improved by transfer of the CCL19 N-terminus to CCL21 (CCL21CCL19-N-term ). Together these results indicate that ligands containing CCL21 core and C-terminal tail (CCL21 and CCL21CCL19-N-term ) are most restricted in their cAMP signaling; a phenotype attributed to a stronger GAG binding of CCL21 and defined structural differences between CCL19 and CCL21.

СУЧАСНІ АСПЕКТИ РОЛІ ГЛІКОЗАМІНОГЛІКАНІВ ЕКСТРАЦЕЛЮЛЯРНОГО МАТРИКСУ У РОЗВИТКУ ГЕНЕРАЛІЗОВАНОГО ПАРОДОНТИТУ ТА ПЕРЕБІГУ ПРОЦЕСІВ РЕПАРАЦІЇ

The prevalence of periodontal disease, in particular of generalized periodontitis, has increased up to 98.5% over the past decades. The topical issue of modern dentistry is to fi nd the cause-effect relations of the development of infl ammatory and dystrophic infl ammatory periodontal diseases. It has been established that sulfated and non-sulfated glycosaminoglycans (GAGs) support the structure of periodontal tissues. The group of sulfated glycosaminoglycans (sGAGs) is represented in the bone tissue of the alveolar ridges by chondroitin sulfates, dermatan sulfate, keratan sulfate and heparan sulfate. Hyaluronic acid, which belongs to non-sulfated glycosaminoglycans, is present in a small amount. The role of biofi lm proteinase is important in the pathogenesis of gingivitis and periodontitis, when the activity of acidic and faintly acid proteinases of dental deposits and gum tissues increases 4-5 times. This process is accompanied by degradation of glycoproteins and other proteins of periodontal tissues. The enzymes of beta-glucuronidase, hyaluronidase, beta-NAcetylhexosaminidase and chondroitin sulfatase are actively involved in the cleavage of acid glycosaminoglycans and glycoproteins of the intercellular substance, periodontal cell membranes, and thus the destruction of circular ligament and periodontal tissues in general. Healing, as a complex dynamic process, is implemented with the inclusion of soluble mediators, blood cells, components of the extracellular matrix and resistant cells involved in recovery and tissue integration. Therefore, the role of GAGs in the processes of periodontal healing is active: 1. Inhibition of synthesis of lipids; 2. Inhibition of activity of proteolytic enzymes; 3. Inhibition of synergistic effect of enzymes and oxygen radicals; 4. Reduction of biosynthesis of infl ammation mediators due to masking of secondary antigenic determinants and inhibition of chemotaxis; 5. Inhibition of apoptosis; 6. Construction of collagen fi bers; 7. Regulation of cell proliferation; 8. Regulation of biosynthesis of the intercellular matrix components; 9. Improvement of microcirculation processes; 10. Rearrangement in structures of proteoglycans; 11. Regulation of chondro- and osteogenesis. The references present a generalized formulation of the main mechanisms of effect of periodontal structures glycosaminoglycans, which is relevant and requires further studying.

The structure of the cutaneous pedal glands in the banded snail Cepaea hortensis (Müller, 1774).

Although gastropods have been crawling through the ocean and on the land for 60 million years, we still know very little about the sticky mucus produced in their foot. Most research has been focused on marine species in particular and, to a lesser extent, on the well-known terrestrial species Arion vulgaris and Cornu aspersum. Within this study, we aim to characterize the foot anatomy of a smaller representative of the family Helicidae, the banded snail Cepaea hortensis. We are particularly interested in the microanatomy of the foot glands, their position, and the histochemical nature of their secretory content. Characterization of the dorsal foot region of Cepaea hortensis reveals four glands, differing in their size and in the granules produced. Histochemically, three of them react positively for sugars (PAS staining and lectin affinity tests for mannose, glucose and N-acetyl-d-glucosamine) and acidic proteins (positive Alcian blue and Toluidine blue staining), indicating the presence of acidic glycosaminoglycans. The fourth gland type does not react to any of these dyes. The ventral pedal region includes two different gland types, which are positive for the presence of acidic glycoproteins, with a lectin affinity for mannose only. A comparison with Helix pomatia indicates differences regarding the number of glands and their contents. In Helix, only three gland types are described in the dorsal region of the foot, which show a similar granular appearance but nevertheless differ in their chemical composition. Congruently, there are two gland types in the ventral region in both species, whereas in Helix an additional sugar moiety is found. This raises the question whether these differences between the pedal glandular systems of both helicid species are the result of protection or size-related adaptations, as they occur in the same habitat.

Effect of sex on histological and histochemical structures of interdigital sinus in adult Bakhtiari sheep of Iran

The aim of this study was to investigate the effect of gender on the histological and histochemical structures of various anatomical regions of the interdigital sinus in the Iranian native sheep. Tissue samples from different anatomical regions of each sinus were obtained from 15 female and 15 male adult Bakhtiari sheep. The sections stained with H&E, Periodic acid- Schiff, Alcian blue, Verhoeff’s, Masson’s trichrome, and Gomori’s method. The sinus was covered by skin and fibrous capsule. The sinus contained descending part, flexure, ascending part, excretory duct and orifice. The sebaceous glands were simple branched acinar and the sweat glands were simple coiled tubular type. The secretory caps were observed in the secretory cells protruded into the lumen. The various histological and histochemical structures of the interdigital sinus showed no considerable differences among various anatomical regions of the right and left sinuses. Also, no significant sex-based differences were found. The surface epithelial cells of the apocrine glands and their secretion contained both neutral and acidic glycosaminoglycans. It is concluded that the general histological and histochemical structures of the interdigital sinus in Bakhtiari sheep were similar to those of other native sheep, but there were also some differences.

Snail mucus − glandular origin and composition in Helix pomatia

Apart from their well-known culinary use, gastropod species such as Helix, which have a hydrogel-like mucus, are increasingly being exploited for cosmetic, bioengineering and medical applications. However, not only are the origin and composition of these “sticky” secretions far from being fully characterized, the number and morphology of the mucus glands involved is also uncertain. This study aims to characterize in detail the cutaneous glands of the Helix pomatia foot on morphological, histochemical and immunohistochemical levels. Hereby the focus is on the gland position and appearance on the foot sole as well as on the chemical nature of the different gland secretions. At least five different gland types can be distinguished by their microanatomy; three are located on the dorsal side and two on the ventral side of the foot sole. Most glands are reactive for acidic proteins and sugars such as mannose and fucose, indicating the presence of acidic glycosaminoglycans. One dorsal gland type shows high reactivity for acidic proteins only. The isolated mucus includes a certain amount of the elements chlorine, potassium and calcium; evidence for lipids was also confirmed in the isolated mucus. The present results for Helix pomatia show a clear difference in the number of glands compared to the related species Helix aspersa (only four mucus glands); histochemically, the glands of both species similarly produce acidic proteins as well as acidic glycosaminoglycans. While calcium ions are known to play a role in mucus formation, the presence and function of other ions such as potassium still need to be clarified.

PATHOMORPHOLOGICAL AND COMPUTERIZED TOMOGRAPHY STUDY OF IBUPROFEN AND COMBINATION WITH 2-PHENYL-3-CARBETHOXY-4-DIMETHYLAMINOMETHYL-5-OKSIBENZOFURAN HYDROCHLORIDE (VINBORON) INFLUENCE ON THE JOINTS OF LOWER EXTREMITIES OF RATS WITH EXPERIMENTAL RHEUMATOID ARTHRITIS

The most frequent adverse reactions to nonsteroidal anti-inflammatory drugs are upper and lower gastrointestinal tract disorders, hepatotoxicity, nephrotoxicity, a decrease in the synthesis of proteoglycans of the cartilaginous tissue and allergic reactions. In our previous studies we have found out that Ukrainian vinboron, the antispasmodic with polytrophic pharmacological effects, can improve the safety profile of the popular non-steroidal antiphlogistics ibuprofen and can also potentiate its anti-inflammatory effect. That fact has become the background to a detailed study of the therapeutic action mechanisms of these drugs when they are used together. Aim of the study. To characterize the influence of vinboron (11 mg/kg, i.g.) on the anti-inflammatory activity of ibuprofen (218 mg/kg, i.g.) when combined, in a model of experimental rheumatoid arthritis according to pathomorphological, morphometric and cone-ray computerized tomography of rats’ hind limbs. Materials and methods . To reproduce the conditions for the administration of ibuprofen corresponding to its clinical use, we selected a model of experimental rheumatoid arthritis-adjuvant arthritis (AA) in rats. On the 28 th day the animals were excluded from the experiment. It was followed by the amputation of the hind limbs at the level of the hocks (art. coxae) and fixation of the material in 10.0% aqueous solution of neutral formalin. The ankle joints were decalcified. The drugs were prepared according to the standard procedure. Microscopy and photographing of histological preparations were carried out with the light microscope OLIMPUS BX 41 at magnifications of 40, 100, 200 and 400 times. The obtained images were processed and morphometry was carried out with the program “Quick PHOTO MICRO 2.3”. The contents of the cellular elements was determined on the basis of the equivalent unit area (1 mm2). Cone-ray computerized tomography was carried out with a digital panoramic cone-ray tomographic scanner “Point 3D Combi 500” (PointNix, South Korea), using a planar sensor with a scanning site of 9 × 12 mm and a radial load of 30-100 μSv. Scanning was carried out with a voltage of 52.0 kV and the electric current of 6.0 mA. For the integral characteristics of the linear dimensions of rats’ hind limbs joints we proposed a formula for calculating the joint volume (Patent of Ukraine No. UA 117538). For the integral characteristics of the dimensions of rats’ hind limbs linear joints we proposed a formula for calculating the joint volume (Patent of Ukraine No. UA 117538). Results and discussion. The use of ibuprofen in combination with vinboron was accompanied by restoration of the structure and function of the cartilage and bone tissues, evidenced by the appearance of acidic and sulfated glycosaminoglycans. The restoration of the bone tissue was realized by the proliferation of the connective tissue and its transformation into cartilage. The synovial membrane was hyperplastic with proliferating synoviocytes of both macrophage and secretory types. In rare cases, the phenomenon of moderate edema and minor focal lymphohistiocytic infiltration persisted. Thus, the use of ibuprofen and ibuprofen in combination with vinboron in rats with adjuvant arthritis resulted in a reduction or absence of inflammation in both the joint and periarticular tissues. The analysis of the results of cone-ray computerized tomography and digital radiography showed that against the background of simultaneous administration of ibuprofen and vinboron there was a tendency to increase the anti-inflammatory effect of the studied antiphlogistics on the rats’ joints affected by adjuvant arthritis. On the 28-th day of the experiment there was the indicatation of decrease of the volume of the hind limb joints by an average of 33.8% compared to the control animals and a near-complete restoration of the width of the joint gaps of the joints examined. It was also determined that vinboron can harmonize the anti-inflammatory effect of ibuprofen when they are used together. Сonclusion. The use of vinboron in combination with ibuprofen leads to a more pronounced pathogenetic anti-inflammatory structural-modifying effect of this antiphlogistics.

Topochemistry, optical anisotropy and FT-IR microspectroscopy of the cocoon of Lithurgus chrysurus (Hymenoptera, Megachilidae)

A previous study has not revealed the participation of a mucous component in the cocoon wall of the solitary bee, Lithurgus chrysurus, differing from the cocoon structure reported for many other bee species. However, uncertainty remains, because only the median and rear zones of this cocoon type have thus far been analyzed. Here, we studied the front zone of this cocoon, searching its components and their organization, to fill this knowledge gap. Topochemical assays, polarization microscopy and Fourier transform-infrared (FT-IR) microspectroscopy were used to study cross sections from L. chrysurus cocoon. Three main layers differing in structural organization were found to compose the cocoon wall. Silk fibroins were assumed to constitute the filamentous threads of the inner and outer layers and the laminar structure of the intermediate layer. Deduced from its topochemical properties and FT-IR spectral signature, a foamy material containing mucin glycoproteins and carboxylated acid glycosaminoglycans was found in the intermediate layer. FT-IR analysis using a Savitzky-Golay 2nd-derivative and absence of linear dichroism and birefringence phenomena suggest that a random-coil secondary structure predominates in the foam component. Co-existence of α-helical and β-sheet conformations is also hypothesized for the fibroin component of this cocoon. It is thus concluded that in addition to fibroin elements, a mucous component, likely contributed by a Malpighian tubule secretion, integrates the composition of the front zone of the cocoon wall of L. chrysurus. In addition, the FT-IR analysis of the inner layer silk of this cocoon suggests significant differences in comparison to the silk fibroins of the silkworm, and some minor spectral differences in comparison to published data on the honeybee silk, with respect to protein secondary structure.

Acid glycosaminoglycan (aGAG) excretion is increased in children with autism spectrum disorder, and it can be controlled by diet.

Autism research continues to receive considerable attention as the options for successful management are limited. The understanding of the autism spectrum disorder (ASD) etiology has now progressed to encompass genetic, epigenetic, neurological, hormonal, and environmental factors that affect outcomes for patients with ASD. Glycosaminoglycans (GAGs) are a family of linear, sulfated polysaccharides that are associated with central nervous system (CNS) development, maintenance, and disorders. Proteoglycans (PG) regulate diverse functions in the central nervous system. Heparan sulfate (HS) and chondroitin sulfate (CS) are two major GAGs present in the PGs of the CNS. As neuroscience advances, biochemical treatments to correct brain chemistry become better defined. Nutrient therapy can be very potent and has minimal to no side effects, since no molecules foreign to the body are needed. Given GAGs are involved in several neurological functions, and that its level can be somewhat modulated by the diet, the present study aimed to evaluate the role of GAGs levels in ASD symptoms. Both tGAG and its different fractions were evaluated in the urine of ASD and healthy control childrens. As levels differed between groups, a second trial was conduted evaluating if diet could reduce tGAG levels and if this in turn decrease ASD symptoms. The present study found that tGAG concentration was significantly higher in the urine of children with ASD compared to healthy control children and this was also evident in all GAG fractions. Within groups (controls and ASD), no gender differences in GAG excretion were found. The use of a 90 days elimination diet (casein-free, special carbohydrates, multivitamin/mineral supplement), had major effects in reducing urinary tGAG excretion in children with ASD.

Efeito da iluminação constante sobre a placenta de ratas: um estudo morfológico, morfométrico e histoquímico

A presente pesquisa analisou o efeito da iluminação constante sobre a estrutura placentária de ratas, abordando parâmetros morfológicos, morfométricos e histoquímicos. Vinte ratas albinas foram submetidas aos seguintes tratamentos: 12hL/12hE por 60 dias, e acasaladas em seguida (GI); ausência de luz por 60 dias, e acasaladas em seguida (GII); iluminação constante por 60 dias, e acasaladas em seguida (GIII); iluminação constante por 60 dias, acasaladas em seguida e tratadas com melatonina (GIV). O estímulo luminoso foi em torno de 400 lux. A melatonina foi administrada na água (400mg/mL de etanol). Os resultados mostraram que histologicamente o GII apresentou vacuolização das células do trofospongio. Morfometricamente, o GIII apresentou camada do labirinto com redução no número de trofoblastos sinciciais e maior vascularização materno-fetal, hiperplasia e hipertrofia das células trofoblásticas gigantes, uma maior média da área total do disco placentário; porém, na camada de trofospongio, as células trofoblásticas e trofoblastos sinciciais não diferiram nos grupos experimentais. As placentas do GIV foram semelhantes às do GI. Histoquimicamente não houve alterações nas fibras colágenas, elásticas, reticulares e glicosaminoglicanas ácidas. Em conclusão, a iluminação constante promove alterações morfológicas e morfométricas na placenta de ratas, podendo acarretar redução funcional e restrições ao crescimento fetal. Essas alterações são abolidas pela reposição de melatonina.