Sulfated Acid Mucopolysaccharides Research Articles

Light and electron microscopic study of the eyelids, conjunctiva-associated lymphoid tissue and lacrimal gland in Bilgorajska Goose (Anser anser).

Normal structure of the accessory organs of the eye is essential for normal eye physiology. Among the most important accessory organs of the eye are the eyelids, the conjunctiva-associated lymphoid tissue (CALT) and the lacrimal gland (LG). The aim of this study was to demonstrate the histological structure of the eyelids and LG by histochemical and ultrastructural analysis. The study was performed on 13 adult female Bilgorajska geese. Eyelid samples were stained with the Alcian blue (AB pH 2.5) and periodic acid-Schiff (PAS) methods. Staining methods used for LG were AB pH 2.5, aldehyde fuchsin (AF), PAS and Hale's dialysed iron (HDI). Within the connective tissue of the eyelids, well-developed, diffuse, CALT follicles were observed, mostly under the conjunctival epithelium. Numerous lymphocytes were present within loose connective tissue. Staining of the eyelids with the PAS method demonstrated the presence of goblet cells of a mucous nature, and AB pH 2.5 staining indicated the presence of sulfated acid mucopolysaccharides. PAS staining of LG revealed the presence of secretory cells containing weakly PAS-positive granules. All epithelial cells of the corpus glandulae and the duct systems reacted positively to AB pH 2.5. HDI staining detected the presence of carboxylated acid mucopolysaccharides. Transmission electron microscopy investigations revealed two types of secretory epithelial cells in LG. Both types of LG cells contained drop-like secretory vesicles of different sizes with low or high electron density in cytoplasm, as well as small and large lipid vacuoles, and numerous small primary lysosomes.

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.

Histological and histochemical features of the small intestine in loggerhead sea turtle hatchlings (Caretta caretta)

Loggerheads (Caretta caretta) are endangered marine turtles and basic research is an essential component of conservation. In an attempt to provide further information in this field, we aimed to identify the microscopic morphological and histochemical features of the small intestine in the loggerhead turtle. Four loggerhead turtle hatchlings were used in this study; due to the loggerhead’s endangered status, this study was performed with dead animals. During necropsy, the cranial and caudal portions of the small intestine were obtained. The samples were embedded in paraffin and stained using the following techniques: Hematoxylin and Eosin, Gomori’s trichrome, Periodic acid-Schiff reactive (PAS), Alcian blue (pH 1.0 and pH 2.5), PAS-Alcian blue, Toluidine blue (pH 4.5 and pH 3.5), Giemsa, colloidal iron, Perl’s blue, Wilder’s impregnation, Methenamine silver, Grimelius and Masson-Fontana. The intestinal mucosa present plicae featuring a pseudostratified columnar epithelium with abundant blood vessels and abundant lymphatic vessels in the submucosa; both of these observations represent the main histological findings. The secretory granules in the goblet cells demonstrated the following characteristics: in the cranial portion, neutral mucopolysaccharides, sulfated acid mucopolysaccharides and mucopolysaccharides with sialic acid were found. In the caudal portion, neutral mucopolysaccharides and mucopolysaccharides with sialic acid were observed. Argentaffin cells were detected along the small intestine, primarily in the cranial portion. There are morphological differences, both histological and histochemical, between the cranial and caudal portion in the small intestine of C. caretta. These characteristics are correlated with possible functions in tissue physiology.

Histological and histochemical features of the small intestine in loggerhead sea turtle hatchlings (Caretta caretta)

Loggerheads ( Caretta caretta ) are endangered marine turtles and basic research is an essential component of conservation. In an attempt to provide further information in this field, we aimed to identify the microscopic morphological and histochemical features of the small intestine in the loggerhead turtle. Four loggerhead turtle hatchlings were used in this study; due to the loggerhead’s endangered status, this study was performed with dead animals. During necropsy, the cranial and caudal portions of the small intestine were obtained. The samples were embedded in paraffin and stained using the following techniques: Hematoxylin and Eosin, Gomori’s trichrome, Periodic acid-Schiff reactive (PAS), Alcian blue (pH 1.0 and pH 2.5), PAS-Alcian blue, Toluidine blue (pH 4.5 and pH 3.5), Giemsa, colloidal iron, Perl’s blue, Wilder’s impregnation, Methenamine silver, Grimelius and Masson-Fontana. The intestinal mucosa present plicae featuring a pseudostratified columnar epithelium with abundant blood vessels and abundant lymphatic vessels in the submucosa; both of these observations represent the main histological findings. The secretory granules in the goblet cells demonstrated the following characteristics: in the cranial portion, neutral mucopolysaccharides, sulfated acid mucopolysaccharides and mucopolysaccharides with sialic acid were found. In the caudal portion, neutral mucopolysaccharides and mucopolysaccharides with sialic acid were observed. Argentaffin cells were detected along the small intestine, primarily in the cranial portion. There are morphological differences, both histological and histochemical, between the cranial and caudal portion in the small intestine of C. caretta. These characteristics are correlated with possible functions in tissue physiology.

Clinical characterization and mutation identification for multiple sulfatase deficiency patients in China

Multiple sulfatase deficiency is a rare autosomal recessively inherited lysosomal storage disorder characterized by the accumulation of sulfated lipids and acid mucopolysaccharides. The aim of this study was to explore the clinical manifestations, enzyme activities and SUMF1 gene mutations in two Chinese patients with multiple sulfatase deficiency. One boy and one girl from two families were studied. Both patients presented with mental retardation, mild coarse facial features, a neurodegenerative course of disease with loss of sensory and motor function after 2 years of age, ichthyosis and skeletal abnormalities (kyphosis or/and scoliosis). Clinical characteristics indicate multiple sulfatase deficiency.Sulfatases activities in blood leucocytes, plasma or cultured fibroblast of the patients were measured.Genomic DNAs were extracted from peripheral blood leukocytes from the patients and their parents. All SUMF1 gene exons and intron-exon boundaries were amplified by PCR and subjected for direct sequencing. In case 1, five sulfatases activities of blood leucocytes and four sulfatases of cultured skin-fibroblasts were analyzed.In case 2, three sulfatases activities of blood leucocytes were tested.Significantly decreased sulfatases activities confirmed the diagnosis of multiple sulfatase deficiency.On SUMF1 gene, c.793_794 insATG (p. P265X)/ c.1045C>T (p.R349W) in case 1 and c.451A>G (p.K151E)/ c.1046G>C (p.R349Q) in case 2 were detected, respectively. Three novel mutations c.793_794insAGT, c.1046G>C and c.451A>G were identified. Multiple sulfatase deficiency usually results in multi-organ damage, especially neurologic, skeletal and skin.Sulfatases assay and SUMF1 gene analysis are necessary for the diagnosis. Two Chinese cases with multiple sulfatase deficiency were firstly reported. Three novel mutations were found.It should be considered that the mutation profile of SUMF1 gene in Chinese patients is different from other populations.

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.

Mucinous Meningioma

An unusual variant of meningioma is described that was characterized by small clusters and strands of meningothelial cells surrounded by abundant mucinous stroma. The unusual appearance of the lesion prompted an initial diagnosis of metastatic mucin-secreting carcinoma (so-called colloid carcinoma) resulting in extensive clinical evaluation in search for a primary. Histochemical studies showed the mucinous material to be composed of strongly sulfated acid mucopolysaccharides rich in hyaluronic acid. Immunohistochemical studies showed strong membrane staining of the tumor cells with epithelial membrane antigen and positive cytoplasmic staining with vimentin antibodies. Ultrastructural examination revealed the characteristic features of meningothelial cells (i.e., abundant long, interdigitating cytoplasmic processes joined by well-developed cell junctions) but failed to demonstrate secretory activity within the neoplastic cells. The prominent mucinous stroma in this case most probably represents a nonspecific reaction of stromal cells to an undetermined stimulus. Mucinous meningioma should be added to the list of morphologic variants of meningioma and should be considered in the differential diagnosis of mucinous lesions in intracranial locations.

A morphological and histochemical study of the gland cells of a cercaria referable to the genus Acanthoparyphium (Trematoda: Echinostomatidae)

A cercaria referable to the genus Acanthoparyphium encountered in Cerithidea cingulata has been named as Cercaria bengalensis II. The present paper is the first report on the glandular complex in this genus. Our studies revealed the presence of different types of gland in the body and tail of Cercaria bengalensis II. Morphologically, three types of gland cell were apparent: penetration, cephalic and cystogenous glands. Histochemically, three types could be discerned as cystogenous glands: dorsal granular, dorsal vacuolated and ventral gland cells. Altogether five types of gland cell occurred. During the intraredial stages, a layer covering the tegument of the cercaria originated from the secretions of the ventral gland cells. In addition to this layer, a second layer underlying it was formed by secretions of the dorsal vacuolated cells. The secretions of the penetration glands contained proteins such as keratin, elastin- and collagen-like substances, and carboxylated and neutral mucopolysaccharides. The cephalic glands contained only keratinized proteins and traces of non-sulphated acid mucopolysaccharides. Dorsal granular cells had the task of secreting keratin, disulphides, sulphydryls, elastin- and collagen-like substances as protein components and also sulphated and carboxylated acid mucopolysaccharides and traces of neutral mucopolysaccharides. Dorsal vacuolated cells behaved like dorsal granular cells, but did not secrete neutral mucopolysaccharides. Disulphide-containing proteins, lipids and phospholipids were also present. The ventral gland cells consisted only of strongly sulphated acid mucopolysaccharides and neutral mucosubstances, with no proteins.

Papular mucinosis in L-tryptophan-induced eosinophilia-myalgia syndrome

Five patients with the L-tryptophan-related eosinophilia-myalgia syndrome had a generalized eruption of flesh-colored papules. In all patients, histologic examination revealed a focal accumulation of mucin in the upper mid dermis, associated with increased dermal cellularity. The mucin was composed predominantly of hyaluronic acid, with small amounts of sulfated acid mucopolysaccharides. The cells within the lesion were fibroblasts. The lesions slowly regressed after L-tryptophan was discontinued. Proposed explanations for the L-tryptophan-related eosinophilia-myalgia syndrome have centered on contaminants, chemically related to L-tryptophan, introduced in the manufacturing process. Tryptophan metabolites have been linked with sclerotic cutaneous diseases but have not been previously implicated in cutaneous mucinoses.

The Pathogenicity, Localization, and Cyst Structure of Echinostomatid Metacercariae (Trematoda) Infecting the Kidneys of the Frogs Rana clamitans and Rana pipiens

Light and scanning electron microscopy were used to examine the localization and pathogenicity of echinostomatid metacercariae infecting the kidneys of leopard frogs, Rana pipiens, and green frogs, Rana clamitans. Cysts occurred predominantly in the ventrolateral renal cortex, and at least some were confined to the lumen of the Bowman's capsules. Each vermiform metacercarial body was enclosed by a spherical cyst wall that had a uniform thickness. The wall was composed of a homogeneous material containing basic and keratinlike proteins, with sulfated acid mucopolysaccharides on the outer surface. Most cysts were enclosed by a fibrous capsule of host origin, or were surrounded by an inflammatory focus. Fibrosis was always focal, but its degree varied between individual hosts and between different cysts within the same host. Some heavily encapsulated cysts were darkened and contained disintegrating worms. In heavily infected kidneys, confluence of fibrotic or inflammatory foci resulted in the displacement of functional renal tissue. These data suggest that infection by echinostomatids may impair renal function and that the host's response affects parasite viability.

The relationship between nitrogen output and changes in mucous cell function in the amphibious teleost Blennius pholis L. during aerial exposure

The numbers of mucous cells in the epidermis of the head and body and in the surface of the gill arch of the amphibious blenny, Blennius pholis L., were estimated on fish immersed in sea water and after 4 h aerial exposure. During emersion there appeared to be a considerable reduction in the frequency of epithelial mucous cells in the areas studied, although counts for the head epidermis were somewhat variable. A concomitant decline in the number of cells thought to be actively secreting was also recorded in tissue samples from both the head and gills, while in the body epidermis the potential for mucus‐secretion was maintained close to the levels observed in immersed fish.Histochemical studies revealed epidermal mucous cells containing either sialylated acid mucopolysaccharides or neutral mucins, or a mixture of these, in both the head and the body, whereas in the gill arch epithelia there were, in addition, cells containing sulphated acid mucopolysaccharides. After emersion, a disproportionate loss of cells containing neutral and sialylated mucus from the gill epithelia resulted in an increase in the proportion of secreting cells staining positively for sulphated acid mucopolysaccharides.The results of this study are discussed in relation to nitrogenous excretion during aerial exposure.

Chondroid syringoma of the upper lip: Report of a case

The case of a 38-year-old white man with a chondroid syringoma in the left side of the upper lip is reported. Histochemical study of the lesion showed that the morphologic distributions of the neutral mucosubstances, sulfated acid mucopolysaccharides, and elastic fibers in the reported case were similar to those of mixed salivary gland tumor. The recently reported ultrastructural and immunohistochemical features of chondroid syringoma are also discussed in an effort to clarify its histogenesis.

Histochemical studies on the vitellogenesis in a fairy shrimpStreptocephalus dichotomus Baird (Crustacea: Anostraca)

Histochemical characterisation of ooplasm and yolk granules in different stages of oocyte maturation in the ovary was made in the freshwater fairy shrimpStreptocephalus dichotomus. In stage I oocyte, the ooplasm is reactive to basic and acidic groups of protein, aromatic groups such as tryptophenyl and tyrosyl and sulphur containing substances. The other organic compounds include acid mucopolysaccharide and acidic lipids. In stage II oocyte, sudanophilic lipoidal substances appear coinciding with yolk formation. Stage III oocytes are characterised by the abundance of yolk granules which are rich in sulphated acid mucopolysaccharide as well as glycogen. Besides these, the presence of acid phosphatase activity in the ooplasm and nurse cell is also noticed and its functional role in relation to vitellogenesis is discussed.

Seasonal changes in the pineal organ of control and gonadectomized fish (Mystus vittatus [Bloch]).

The surface area and luminal structures of the pineal sac were studied during the prespawning, spawning, and postspawning phases of control and gonadectomized fish, Mystus vittatus, throughout the year. From the later half of prespawning to the first half of postspawning phases the values for the surface area were lower in the gonadectomized group whereas from the later half of the postspawning to the first half of prespawning phases the values were higher than in the control group. The acidophilic cell bodies were visible in the first half of prespawning and at the end of spawning phases as well as during the postspawning phase. In the control group sulfated acid mucopolysaccharides and mucoid basophils with gamma substances were faintly stained during the early months of the reproductive cycle in contrast to the experimental group; but in the later months these substances were strongly stained in both groups except in the gonadectomized group at the end of postspawning phase. The pineal lumen was narrow during the pre- and postspawning phases and wider in the spawning phase of the gonadectomized group in comparison to control; this may be related to the flow of cerebrospinal fluid. Thus, the morphohistochemical changes in the pineal organ of fish were evident in both groups during the prespawning, spawning, and postspawning phases.

67Ga-binding substances in stomach, small intestine, pancreas, and muscle.

Normal male rats were injected with either gallium citrate Ga 67 or sodium sulfate S 35. After 24 h, the stomach, small intestine, pancreas, and muscle were excised and homogenized. After the removal of the nuclear fraction, each of these homogenates was digested with protease. After digestion, the supernatants of the reaction mixtures were applied to a Sephadex-G-100 column. The radioactivity was eluted with buffer solution. The resultant eluates were analyzed for radioactivity and the levels of proteins, uronic acids, and sialic acids. In all four organs, sizable amounts of 67Ga were bound to sulfated acid mucopolysaccharides with molecular masses of about 10,000 daltons and to sulfated acid mucopolysaccharides, a species whose molecular masses exceed 40,000 daltons. In the stomach, large amounts of 67Ga were bound to sulfated acid mucopolysaccharides with molecular masses of about 10,000 daltons. From these results, it is obvious that the main 67Ga-binding substances in these four organs are sulfated acid mucopolysaccharides, and that these acid mucopolysaccharides play the most important role in the concentration of 67Ga in these organs.

Differential cellular response of resistant and susceptible rodents to the early stages of infection of Taenia taeniaeformis

Taenia taeniaeformis infection course in C3H He and BALB/c mice (susceptible and resistant respectively to T. taeniaeformis) was compared for 10 days post-infection (p.i.). Light microscopic observation of early organisms in BALB/c revealed host cell infiltration as early as 2 days p.i. By day 4 p.i. the larvae presented an inconspicuous acellular halo and were surrounded by a dense mass of leukocytes, mainly eosinophils. The amorphous substance reacted positive for sulphated acid mucopolysaccharides. By day 6 p.i. the parasite-host interface was filled by leukocytes and fibroblasts, staining with alcian blue was limited to small areas and the parasites appeared disrupted and lightly stained by haematoxylin-eosin. At day 10 p.i. the parasite had been destroyed by the host. In rats and C3H He mice, the parasites at day 2 p.i. were 10 μm in diameter, bounded by a larger amorphous acellular halo that reacted positive to alcian blue stain, an indication of the presence of acidic muco-polysaccharides. By day 4 p.i. the clear halo became larger and reacted strongly positive for acid mucopolysaccharides particularly close to the organism. By day 6 p.i. alcian blue positive material appeared more diffuse, covering a broad area well inside the host tissue. By this time a layer of inflammatory cells were often present at some distance from the parasite tegument. By day 10 p.i. the larvae continued growing and increasing the number of tegumental cells. The presence of sulfated acid mucopoly-saccharides around the larvae probably serves as a protective barrier for the parasite by triggering the alternative pathway of complement, consuming C3 and C5, thus preventing cell attachment and humoral antibody attack. The early destruction of the parasite by cell defense mechanism implies a major role of cellular immunity in T. taeniaeformis elimination.

67Ga-binding substances in abscess and normal tissues.

Abscess-induced animals and normal animals were treated with 67Ga-citrate. Abscess, kidney, heart, lung, and spleen were excised and homogenized. After removal of the nuclear fraction, each of these homogenates was digested with protease. After digestion, the supernatants of the reaction mixtures were applied to a Sephadex G-100 column. Resultant eluates were analyzed for radioactivity, protein, uronic acids, and sialic acids. Sodium sulfate-35S was administered to animals that were then treated by the same procedure as that followed for animals treated with 67Ga-citrate. In abscess, kidney, lung, heart, and spleen, sizeable amounts of 67Ga had been bound to the sulfated acid mucopolysaccharides with molecular weights of about 10,000, and to the sulfated acid mucopolysaccharides, a species whose molecular weights exceeded 40,000. Based on the results presented here, it is clear that 67Ga-binding substances in abscess and also in the above four organs are sulfated acid mucopolysaccharides.

Changes in the concentration and histochemistry of epidermal mucous cells during the alevin and fry stages of the Brown trout Salmo trutta

Changes in the concentration and histochemistry of epidermal mucous cells have been shown to be similar in both natural and hatchery‐reared Brown trout during the early stages of development. Superficial mucous (goblet) cell concentration is greatest immediately post‐hatching (≈ 1500 cells/mm2) and remains high in the alevin epidermis during the period of yolk‐sac resorption. Once the yolk sac has been resorbed and the free‐swimming fry have left the spawning redds, the epidermal mucous cell concentration falls to ≈700 cells/mm2. During the alevin‐fry transition there is also a progressive decrease in the relative proportion of mucous cells containing sulphated (or a mixture of sulphated and carboxylated) acid mucopolysaccharides and an increase in the proportion of cells containing exclusively carboxylated acid mucopolysaccharides. These findings are discussed in relation to habitat and to the functional roles of epidermal mucous secretion in fishes. It is suggested that the high potential for production of mucus in the alevin stages is related to the need for protection against abrasion during the period of residence in the gravel of the spawning redds.

EARLY POST-METAMORPHIC GROWTH, BUDDING AND SPICULE FORMATION IN THE COMPOUND ASCIDIAN CYSTODYTES LOBATUS.

1. The colonial ascidian Cystodytes lobatus has a long breeding season (at least 6 months) and releases tadpoles sporadically throughout the day, indicating a long period of recruitment. 2. Tadpoles of C. lobatus were settled and reared in the laboratory in order to observe early growth, budding and spicule formation. 3. Budding is preceded by the formation of a new stomach at the posterior end of the esophagus and fits Nakauchi's Type I budding pattern. 4. Spicule formation begins within 5 days after settlement. The spicules appear to form in a particular region at the anterior end of the abdomen and migrate over the abdomen to form a single or slightly overlapping layer embedded in a "tunic spicular lamina." This lamina lies between the common tunic and the zooid cavity and forms a spicular sac in the tunic surrounding but separated from the abdomen of each zooid. It stains especially heavily for sulfated acid mucopolysaccharide; the spicules are concluded to form by cellular action in this organic matrix. 5. There is great variation among zooids in the quantity of spicules formed. These differences are maintained in the buds, resulting in colonies in which all zooids either have few or many spicules, and are therefore probably genetic in origin. 6. During budding the spicular sac becomes disrupted and appears to bud, resulting in a reallocation of the spicules to the buds and formation of separate spicular sacs around the abdomen of each bud. At budding there is apparently no disruption of bladder cell membranes in the tunic and no dissolution of spicules by the acids contained in the bladder cells.

Local responses in primary and secondary human lung cancers. I. Patterns of cellular (eosinophils and macrophages) and extracellular (acid mucopolysaccharide) reactions

Local free cell (eosinophils and macrophages) and extracellular (acid mucopolysaccharides) reactions were studied by histochemical techniques in 72 primary lung cancers (Group A), 17 pulmonary metastases and 5 lung tumours of unknown origin (Group B). Strong cellular reactions were more frequent in Group A than in Group B, whereas extracellular reactions were more frequent in Group B than in Group A. In both groups the degrees of eosinophilic infiltration and accumulation of acid mucopolysaccharides tended to be negatively correlated. In contrast, macrophages and eosinophils showed a positive correlation in the primary cancers, but none in the other tumours. The extracellular material had physical and chemical properties (solubility, affinity to stains and degradability by enzymes) of the sulphated acid mucopolysaccharides. From its distribution it appeared to be derived from fibroblasts and/or tumour cells.