Sulfated Mucopolysaccharides Research Articles

Pentachrome Staining: A Comprehensive Review of the Novel Method for Collagen and Sulfated Mucopolysaccharides

Pentachrome Staining: A Comprehensive Review of the Novel Method for Collagen and Sulfated Mucopolysaccharides

In vitro effects of λ-carrageenin in the head-kidney leucocytes of gilthead seabream (Sparus aurata)

The λ-carrageenin is a sulphated mucopolysaccharide that has been used for decades to induce experimental inflammation in mammals. However, it has been little considered in fish. We studied the in vitro effects of λ-carrageenin on gilthead seabream (Sparus aurata L.) head-kidney leucocytes (HKLs). For this purpose, HKLs were incubated with serial concentrations (from 0 to 1,000 μg mL-1) of λ-carrageenin for 3, 6, 12 and 24 h to assess its influence on cell viability and morphology, cell activity and modulation of several selected inflammation-related genes. The viability results demonstrated that λ-carrageenin has no negative effects on HKLs. The respiratory burst activity and phagocytic ability of HKLs after being incubated with λ-carrageenin (100 and 1,000 μg mL−1) for 24 h were increased, whereas the phagocytic capacity was inhibited by the higher dose at the same experimental time compared with control samples. However, the peroxidase activity of HKLs was not changed by incubation with λ-carrageenin. According to transmission electron microscopy results, incubation of HKLs with the higher dose of λ-carrageenin appeared to activate the cells being evident different morphological changes without sign of cell death. Furthermore, up-regulation of three proinflammatory cytokines (il1b, tnfa, and il6) and down-regulation of anti-inflammatory genes (tgfb) were denoted in HKLs incubated with carrageenin. The present results provide a detailed approach to the effects of λ-carrageenin on fish leucocytes, which could have some impact on how we understand the response of these cells when inducing an inflammatory process in fish.

Mucopolysaccharide from cuttlefish: Purification, chemical characterization and bioactive potential

The sulfated mucopolysaccharide (GAG) was isolated from S. pharonis and the carbohydrate and protein content was found to be 62.4% and 3.9%. The disaccharide profile of sulfated GAG composed glucuronic acid, N-acetyl glucosamine and sulfate content by contributing 50.11%, 38.00% and 27.69% respectively. The carbon, hydrogen and nitrogen content of the sulfated GAG showed 14.80%, 1.68% and 2.99% respectively. The molecular weight of sulfated GAG was calculated as 27kDa and the structural characterization was done by Fourier Transform Infrared (FT-IR) and NMR Spectroscopy. The Activated Partial Thromboplastin Time (APTT) and Prothrombin Time (PT) of sulfated GAG were determined as 91 IU and 39.55 IU at 25μg/ml respectively. Further the sulfated GAG reported the cytotoxic effect (CC50) of 1100μg/ml concentration on Vero cell line. The sulfated GAG reported the anticancer activity against HeLa cell line with an inhibition rate of 18.65%–66.13% at 50–250μg/ml concentration. The sulfated GAG can be considered as a potent anticoagulant and anticancer drug in future.

Clasper gland morphology and development in Potamotrygon magdalenae (Elasmobranchii: Potamotrygonidae).

Clasper gland morphology and development in Potamotrygon magdalenae and its relation with the acquisition of reproductive maturity is described in males of different developmental stages (embryos, neonates, juveniles, and reproductively active and resting adults). The glands are subcutaneous masses in the proximal base of each clasper. They are partially bilobate organs with a ventral groove that bears a row of papillae. Glands tend to be asymmetric, the left gland has a larger size, a trend that has been observed in other organs of elasmobranchs. Glands are formed by radially organized tubular secretory units lined with a simple columnar epithelium with basal nuclei and granular eosinophilic cytoplasm; vascularized loose connective tissue surrounds the gland units. The gland is covered by two layers of striated muscle tissue in circular and longitudinal arrangement. The clasper glands begin to develop in neonates and their secretory activity begins in juveniles. The active secretion of the clasper gland is observed in mature males, it includes glycoproteins and sulfated mucopolysaccharides. The size of the glands has a positive and direct relationship with body size, measured as disc width. Significant differences in clasper gland size were found between mature (active and resting) and immature (neonates and juveniles) males, suggesting that the acquisition of the sexual maturity involves the increase in the size of the gland due to a highly augmented secretory activity. Therefore, clasper glands are clearly associated with the reproductive activity of males and their secretion should have an endocrine control as other sexual secondary organs. J. Morphol. 278:369-379, 2017. © 2017 Wiley Periodicals, Inc.

Pharmacognostical and Histochemical Studies on Apakva Kadali (Unripe Banana Fruit)

The unripe fruit of Musa × paradisiaca L., classically known as apakva kadali, belonging to the family Musaceae, is used as pa- thya in atisara (diarrhea), in grahani (irritable bowel syndrome), and agnimandya (loss of appetite). The unripe fruits are considered to be helpful in the management of diabetes mellitus. Being an important medicinal plant, the present investigation is an attempt to explore the pharmacognostical analysis and thin-layer chromatography fingerprint studies on the widely used Musa. × paradisiaca L. The studies revealed that unripe banana fruit showed the presence of small outer protrusion of papillae on the epidermis, abundant tannin-containing cells in the epicarp and mesocarp. Elongated air-spaced cells are surrounded by the parenchyma cells in the epicarp, abundant starch grains in the mesocarp portion, and acicular crystals of calcium oxalate in raphide bundles. To detect the location of various constituents of the drug, sections of unripe fruits were treated with various reagents, and studies showed the presence of phenols, protein, calcium oxalate, magnesium, starch, cellulose, calcium pectate, lignin, sulfated mucopolysaccharides, tannin, and silica contents. Phytochemical studies showed the presence of alkaloids, carbohydrates, phenols, proteins, starch, tannins, steroids, and saponins.

Structure and Function of the Reproductive System of Aplysia kurodai

This study investigated structure and function of the reproductive system in Aplysia kurodai by means of anatomical, histological, and histochemical observation. Reproductive system of this species is consisted of ovotestis, small hermaphroditic duct, ampulla, accessory genital mass and large hermaphroditic duct. The ovotestis is composed of a large number of follicles, and both oocytes and spermatocytes matured in the same follicle. The small hermaphroditic duct is a single tube and contains a swelling, the ampulla, which functions as a storage organ for endogenous sperm and an oviduct. The accessory genital mass is connected to both the small and large hermaphroditic duct, and consisted of three glands: albumen, membrane (winding) and mucus gland. The albumen gland is consisted of granular cells producing basophilic and neutral mucopolysaccharides. The membrane and mucus gland are consisted of granular cells producing acidophilc and sulfated mucopolysaccharides. The large hermaphroditic duct is a single tubular gonoduct linking the accessory genital mass to the common genital aperture but is consisted of two parallel compartments. Internally, these two compartments are incompletely divided by internal septum or fold, which are called as the red hemiduct and white hemiduct, respectively. The red hemiduct functions as an oviduct and the white hemiduct functions as a copulatory duct. The reproductive system of A. kurodai is externally comprised a single tube, i.e., monaulic type. However, internal structure of duct is incompletely divided into oviduct and copulatory duct, i.e., the oodiaulic type.

Pretreatment and Integrated Analysis of Spectral Data Reveal Seaweed Similarities Based on Chemical Diversity

Extracting useful information from high dimensionality and large data sets is a major challenge for data-driven approaches. The present study was aimed at developing novel integrated analytical strategies for comprehensively characterizing seaweed similarities based on chemical diversity. The chemical compositions of 107 seaweed and 2 seagrass samples were analyzed using multiple techniques, including Fourier transform infrared (FT-IR) and solid- and solution-state nuclear magnetic resonance (NMR) spectroscopy, thermogravimetry-differential thermal analysis (TG-DTA), inductively coupled plasma-optical emission spectrometry (ICP-OES), CHNS/O total elemental analysis, and isotope ratio mass spectrometry (IR-MS). The spectral data were preprocessed using non-negative matrix factorization (NMF) and NMF combined with multivariate curve resolution-alternating least-squares (MCR-ALS) methods in order to separate individual component information from the overlapping and/or broad spectral peaks. Integrated analysis of the preprocessed chemical data demonstrated distinct discrimination of differential seaweed species. Further network analysis revealed a close correlation between the heavy metal elements and characteristic components of brown algae, such as cellulose, alginic acid, and sulfated mucopolysaccharides, providing a componential basis for its metal-sorbing potential. These results suggest that this integrated analytical strategy is useful for extracting and identifying the chemical characteristics of diverse seaweeds based on large chemical data sets, particularly complicated overlapping spectral data.

Histochemical Analysis of Glycoconjugates in the Lacrimal Gland of the Camel ( Camelusdromedarius )

The mammalian lacrimal gland secretes an aqueous solution in which different chemical substances are present, including proteins and carbohydrates. The histochemical investigations on characterization of camel lacrimal secretion are limited. This study aimed to determine the histochemical properties of the dromedary camel lacrimal gland. Glandular samples of twenty healthy adult camels of both sexes were collected at Omdurman slaughterhouse, Sudan. The camel lacrimal gland was compound with tubulo-acinar mixed secretory units, having serous and mucous cells. Best’s carmine reactionwas negative, whereas a strong positive reaction to periodic acid-Schiff's (PAS) diastase resistant was observed in most of the secretory units; the reactions to alcian blue (AB pH 2.5) and aldehyde fuchsin (AF) were strong in the glandular tubules and weak or negative in the acini. Goblet cells in the epithelium of the interlobular ducts were strongly PAS, AB pH2.5 and AF positive.Both acini and tubules showed positive reactions to acid and alkaline phosphatases especially in their apical parts. These results suggest that the predominant acini secretion was serous which contained mainly neutral mucopolysaccharides, whereas the tubules secretion was mucous thatcontained sulfated and carboxylated acid mucopolysaccharides. Moreover, the different staining properties of the camel lacrimal secretion reflect various physiological functions.

Histology and Histochemistry of Mantle of Lymnaea Luteola (Lamarck 1799) Mollusca Gastropoda

The mantle is an important molluscan organ. It serves as a respiratory organ in most of the pulmonates; its walls have a dense vascular network. It is an outer jacket of the visceral mass and harbors the pulmonary chamber on its right side. Mantle is the specialized organ of the gastropods for it not only bears the glands that secrete the shell but also acts as buffer between the hard calcareous shell and the animal itself, spreading like a cushion through the entire inner surface of the body whorl. In Lymnaea luteola two types of gland cells are noticed in the mantle;epithelia and sub - epithelial. The epithelial mucous glands secrete metachromatic, alcianophilic sulphated mucopolysaccharides. The sub - epithelial mucous glands are classified into two categories, according to their histochemical reactions. Some of them secrete non - metachromatic, alcianophilic sulphated mucopolysaccharides and others secrete mucosubstances which are carboxylated.

Histological and certain histochemical observations on the labial salivary glands of the buffalo , bubalus bubalis.

Histological, morphometric and histochemical investigations were carried out on the labial salivary glands of the buffalo. Both the upper and lower labial glands of the buffalo are branched tubuloacinar type and composed of serous acini surrounded by myoepithelial cells. Histochemically it showed seromucous character, they secrete neutral mucopolysaccharides, mucopolysaccharides acid, and few sulfated mucopolysaccharides. The labial salivary glands of buffalo posses ductal system of both the intralobular ductules and interlobular ducts.

Histomorphology of the hypobranchial gland in Dicathais orbita (Gmelin, 1791) (Neogastropoda: Muricidae)

Tyrian purple is a dye used in antiquity and is a natural product of marine molluscs of the family Muricidae. Bioactive precursors of Tyrian purple occur in the hypobranchial gland, reproductive glands and egg masses of muricids, such as Dicathais orbita. Histomorphological examination of the hypobranchial–gonoduct complex (rectum and hypobranchial, capsule, albumen and rectal glands) was conducted to provide the first description of the hypobranchial gland in D. orbita and to determine a mechanism for the transfer of Tyrian purple precursors to the gonoduct and ultimately to the egg masses. Seven secretory cell types were identified in the hypobranchial epithelium of D. orbita, which can be broadly classified into cells containing mucoproteins and acidic sulphated mucopolysaccharides. Three secretory cells new to the Muricidae were identified, along with two cell types that appear to be associated with synthesis of Tyrian purple. A subepithelial vascular sinus surrounding the rectum and rectal gland occurs between the hypobranchial gland and gonoduct. Examination of this region failed to reveal a direct anatomical mechanism for the transfer of precursors to the gonoduct. However, biochemical similarities in secretions from the hypobranchial, capsule and albumen glands suggest that the synthesis of precursors within the gonoduct may be possible.

Antimicrobial Activity of Sulfated Mucopolysaccharides [Heparin and Heparin-Like Glycosaminoglycans (GAGs)] from Cuttlefish Euprymna berryi Sasaki, 1929

Antimicrobial Activity of Sulfated Mucopolysaccharides [Heparin and Heparin-Like Glycosaminoglycans (GAGs)] from Cuttlefish Euprymna berryi Sasaki, 1929

Heparin-induced thrombocytopenia: Impact of bovine versus porcine heparin in HIT pathogenesis

Heparin-induced thrombocytopenia (HIT) is a complication of heparin therapy in cardiovascular/hematologic indications. Heparin is a mixture of sulfated mucopolysaccharide with heterogeneity and is capable of forming multiple complexes with platelet factor 4 (PF4), released from activated platelets. HPF4 antibodies may cause platelet/endothelial cell activation to promote HIT pathogenesis. HIT is a clinico-pathologic syndrome and its diagnosis primarily remains a clinical one; however, the serologic confirmation of the presence of HPF4 antibodies is also necessary part of the evaluation. Assays are based on the immunodetection of HPF4 antibodies and/or their functional ability to activate cells. Currently, there are several assays in use and a few newer/rapid immunoassays are becoming available. Recent studies have confirmed that HPF4 antibody generation (seroconversion) is common after cardiac surgery and suggest that patients receiving bovine heparin are more likely to generate functional (pathogenic) HPF4 antibodies of the IgG subclass. Thus, the use of bovine heparin in cardiovascular surgery should be avoided. A brief account of the currently available options for the management of HIT patients with non-heparin anticoagulants is provided.

Pathological and histochemical changes in the lungs of rabbits experimentally infected with P.seudomonas aeruginosa

The main objective to this study is to demonstrate certain aspects of pathological and histochomical changes in the lungs of rabbits experimentaly infected with P.aeruginosa . 
 Twelve white Newzoland rabbits of 12-16 weeks old were used . they were randomaly divided into two groups. The first group (6 animals ) were given single intraperitoneal, dose of 108 CFU of P. aeruginosa . The second group (6 animals ) maintained as a control and was inoculated with 1 ml of sterial normal saline by the same route of the first group. All the animals were sacrificed on day three post – inoculation. 
 The pathological exmination showed acute suppurative reaction in pulmonary tissue together with lymphocytic aggregation around airways and blood vessels. The histochomical study revealed that the granules of goblet cells and submucesal glands of the lung in normal rabbits were of neutral mucopolysaecharides and little sialomucin, these substances changed to sulfated acid mucopolysaccharide and sialomucin in the infected animals.

Dystrophic scrotal calcinosis originating from benign eccrine epithelial cysts

Scrotal calcinosis has been classified as a form of idiopathic calcinosis cutis. However, the pathogenesis of the calcified nodules has not been fully elucidated: it is still unclear whether the condition is truly idiopathic, or the result of breakdown of calcified epithelial cysts. We describe a 29-year-old Japanese patient with scrotal calcinosis originating from epithelial cysts. Light microscopy revealed a large epithelial cyst containing von Kossa-positive material and several small dilated ductal structures beside the cyst. The epithelia of the cyst and ductal structures were connected, showing similar eccrine duct differentiation on immunohistochemical staining and electron microscopy. In the cyst lumen, calcium was present as needle-shaped crystals. The pathogenetic mechanism of calcium deposition seemed to be due to excessive production and discharge of matrical debris and sulphated mucopolysaccharides, which derived from luminal cells, and their accumulation in the lumina.

Complementary DNA Cloning and Characterization of Pearlin, a New Class of Matrix Protein in the Nacreous Layer of Oyster Pearls.

Calcified shell layer is composed of two polymorphs of CaCO(3), aragonite or calcite, and an organic matrix. The organic matrix consists of EDTA-soluble and insoluble fractions. These fractions are thought to regulate the formation of the elaborate shell structure. After decalcification of powdered pearl with 0.3 M EDTA, an EDTA-insoluble fraction was extracted with 0.3 M EDTA/8 M urea. This extraction step enabled us to purify a new class of EDTA-insoluble protein, Pearlin, almost homogeneously. Pearlin has a molecular weight of about 15 kDa and contains a sulfated mucopolysaccharide. We cloned the complementary DNA coding for Pearlin and deduced its complete amino acid sequence. Sequence analysis reveals that Pearlin is composed of 129 amino acids with a high proportion of Gly (10.8%), Tyr (10.0%), Cys (8.5%), Asn (7.7%), Asp (7.7%), and Arg (7.7%). Northern blot analysis showed that Pearlin messenger RNA was expressed specifically in mantle epithelium. From the sequencing data, Pearlin was shown to be quite different from the fibrous protein rich in Ala and Gly. The function of this protein in biomineralization is discussed.

Hirudin-based anticoagulant strategies for patients with suspected heparin-induced thrombocytopenia undergoing percutaneous coronary interventions and bypass grafting.

Heparin-induced thrombocytopenia (HIT) is an immune-mediated adverse drug reaction that is associated with thrombotic events of the venous and arterial circulatory systems stemming from an intense and well-characterized prothrombotic triad of platelet activation, coagulation cascade stimulation and vascular endothelial cell injury. Although heparin (or other sulfated mucopolysaccharide compound) cessation represents a vital first step in management, patients remain susceptible to life-threatening thrombosis for up to several weeks, providing a strong rationale for a 'proactive approach' to care that includes prompt initiation of an alternative anticoagulant strategy throughout the high-risk period. The importance of alternative options for anticoagulation is most evident in clinical situations wherein treatment is a recognized standard of care and prerequisite for an optimal outcome. The following review highlights the use of recombinant hirudin (lepirudin) among patients with suspected HIT requiring precutaneous coronary interventions (PCI) and coronary arterial bypass grafting.

Functional heterogeneity of antiheparin-platelet factor 4 antibodies: implications in the pathogenesis of the HIT syndrome.

Heparin-induced thrombocytopenia represents one of the most severe drug-induced disorders of platelets. This syndrome is believed to be mediated through antibodies generated against a heparin-platelet factor 4 complex. Complexation of a sulfated mucopolysaccharide chain of heparin with a platelet granular protein (platelet factor 4) produces an allosteric modification of platelet factor 4 resulting in neoepitope formation and the generation of antiheparin-platelet factor 4 antibodies. These antibodies are capable of activating platelets by binding to heparin, platelet factor 4 and the Fc receptor on platelets, resulting in a complex pathophysiology involving ischemic, thrombotic, and inflammatory processes. To characterize this antibody, IgG fractions were obtained from the serum of patients with heparin-induced thrombocytopenia using ammonium sulphate precipitation and heparin-platelet factor 4-sepharose 4B affinity chromatography methods. With the affinity purification, two major components, peaks I and II, with high antiheparin-platelet factor 4 antibody titers were eluted. The purity of all the fractionated immunoglobulins was established by sodium dodecylsulphate-polyacrylamide gel electrophoretic analyses. While peak I did not induce 14C-serotonin release from platelets in the heparin-dependent assay for heparin-induced thrombocytopenia antibodies (14C-serotonin release assay), peak II and the IgGs obtained with the ammonium sulphate precipitation method exhibited a strong and concentration-dependent activation in the presence and absence of heparin and low molecular weight heparin. These immunoglobulins were treated with heparinase, a cationic ion-exchange resin (Heparsorb), or dialyzed to remove traces of heparin, and when tested in the 14C-serotonin release assay, showed the same high degree of activity. These data are suggestive of the generation of heparin-induced thrombocytopenia antibodies capable of activating platelets directly in a nonheparin-dependent manner. These observations underscore the complex pathophysiology of heparin-induced thrombocytopenia syndrome and suggest that the severity of this syndrome in some patients may be due to the generation of "super-active" heparin-induced thrombocytopenia antibodies capable of activating platelets without the requirement of heparin. This could explain why the cessation of heparin in patients does not necessarily correct the symptoms of heparin-induced thrombocytopenia or associated thrombosis.

Urine analysis in the diagnosis of mucopolysaccharide disorders.

The mucopolysaccharidoses are a rare group of inherited disorders of connective tissue metabolism characterized by the storage of sulphated mucopolysaccharides (MPS). The phenotypic features are often apparent early in life: dysmorphic or coarse facial features, hirsutism, skeletal abnormalities, joint stiffness and developmental delay. Inheritance is autosomal recessive except for Hunter syndrome which is Xlinked. The first clinical description of patients with MPS diseases was made in 1917 by Charles Hunter.' He described two male siblings with a unique facial appearance (gargoylism), hepatosplenomegaly, short stature, deafness, heart disease and joint stiffness. In 1919 Gertrude H urler? described two unrelated male patients with the same features but with the addition of corneal clouding and mental retardation. These disorders became known as mucopolysaccharidoses in 1952 following the discovery by Brante' of the presence of chondroitin sulphate-like substances stored in the liver and meninges of a Hurler patient. In 1957 Dorfman and Lorincz reported increased excretion of MPS or glycosaminoglycans (GAGs) in the urine of patients suffering from a mucopolysaccharidosis. This proved to be of great diagnostic importance and led to the development of urine screening tests for the mucopolysaccharidoses. The type and ratio of the urinary MPS were found to be indicative of the subclass of MPS disease present. The mucopolysaccharidoses were shown to be lysosomal storage diseases by Van Hoof and

1007 Early development of primary vestibulocerebellar axons in the rat embryo

Comparison of the [35S]mucopolysaccharides extracted after in vitro incubation of skin biopsy specimens from nonlesional and lesional sites of a patient with mastocytosis showed that lesional sites incorporated sulfate into heparin. After in vitro incorporation of the [35S]sulfate, the tissues were extracted sequentially by a 3-step procedure which utilized high salt concentrations, enzymatic digestion and base hydrolysis to liberate essentially all the counts. The extracted [35S]mucopolysaccharides were separated from free [35S]sulfate, histamine, protein, and hyaluronic acid by ion-exchange chromatography utilizing Dowex 1. The [35S]mucopolysaccharide extracts of the nonlesional skin were completely degraded by treatment with chondroitinase ABC, as they are predominantly dermatan sulfate with small amounts of chondroitin sulfates. The absolute quantity of sulfated mucopolysaccharides after Dowex 1 chromatography in micrograms of uronic acid per mg wet weight of starting tissue was higher in the lesional than the non-lesional specimen, while the specific incorporation of [35S]sulfate per microgram of uronic acid was the same. Approximately one-half of the [35S]mucopolysaccharides obtained in the 3 sequential extracts of lesional tissue was resistant to degradation by chondroitinase ABC as determined by gel filtration before and after enzyme treatment, indicating the presence of sulfated mucopolysaccharides in addition to chondroitin and dermatan sulfates. Heparinase treatment of the chondroitinase ABC-resistant [35S]mucopolysaccharides followed by gel filtration revealed an equal distribution of label between heparin and heparinase-resistant material presumed to be heparan sulfate. Heparin was also directly demonstrated in extracts of lesional mastocytosis skin by chemical and functional criteria.