Acid Mucopolysaccharide Content Research Articles

Model of Vibrio cholerae biofilm as a mechanism of its survival in surface water reservoirs

Vibrio cholerae eltor strains with different epidemic importance isolated from river water in the city of Vladivostok during a cholera outbreak (1999) and in the city of Irkutsk during a safe cholera period (2005) are used in the experiment. A biofilm structure consisting of a peripheral part, bundles, polysaccharide matrix, canals, and polymorphic vibrios is presented by light and luminescent microscopy. The metachromatic pink coloring of the matrix (crystal violet and toluidine blue) or fluorescent reddish orange color (acridine orange) are evidence of acid mucopolysaccharide (glucosaminoglycans) content. The biofilm of a toxigenic strain as opposed to a nontoxigenic one is formed much later, while the elements comprising its structure are more apparent. The viability of vibrio cells during the experiment (90 days) preserving the initial pathogenic potential testifies to the highly adaptable properties of the Vibrio cholerae eltor, which promotes its survival and existence in surface water reservoirs under favorable ecological conditions (optimal temperature, existence of chitin-containing substratum, etc.).

Electrohydrodynamic drying of sea cucumber (Stichopus japonicus)

To develop an improved dehydration method for sea cucumber, and reduce energy consumption, we dried sea cucumber using three different drying methods, namely, electrohydrodynamic (EHD) drying at 18 °C, ambient air drying at 18 °C and oven drying at 80 °C. The quality parameters of the dried products such as shrinkage rate, water absorption, rehydration ratio, protein and acid mucopolysaccharide contents, and texture and sensory characteristics were investigated. Energy consumption in the three drying methods was also examined. Based on the experimental results, oven drying took the shortest time to complete but presented poor quality and needed high energy consumption. EHD drying generated the best quality and consumed only 21.31% of the electric energy required for oven drying, but took a longer time to complete. Meanwhile, ambient air drying yielded indeterminate quality and did not consume electric energy but took the longest time to complete. Considering the requirements for products quality and energy conservation, EHD proved to be the best drying method.

Water content, body weight and acid mucopolysaccharides, hyaluronidase and β-glucuronidase in response to aestivation in Australian desert frogs

This study investigates the effects of aestivation on body water content, body mass, acid mucopolysaccharide (AMPS) and some of its degrading enzymes in different tissues for some Australian desert frogs. The AMPS component of the liver, kidney, skin and cocoon alter during aestivation to help retain water, which is unchanged in most tissues of all frog species, and to protect the frogs from desiccation during extended periods of aestivation. Hepatic AMPS was unaltered in Cyclorana maini, C. platycephala and Neobatrachus sutor but increased significantly after 2 months of aestivation in C. australis. The level of AMPS in the kidney was elevated in all four frog species after 5 months of aestivation. Skin AMPS content in the skin of awake frogs decreases with aestivation period and increases in the cocoon. AMPS in the cocoon probably works as a cement between the cocoons’ layers and its physical presence presumably contributes to preventing water flux. Changes in AMPS content in different tissues were accompanied by significant changes in both hyaluronidase and β-glucuronidase activities, which play an important role in AMPS metabolism. Alcian blue staining of control and digested skin of C. australis and C. platycephala with testicular hyaluronidase indicated the presence of AMPS, concentrated in a thin layer (called ground substance, GS) located between stratum compactum and stratum spongiosum, and acid mucin concentrated in the mucous glands and in a ‘tubular’ structure which could be observed in the epidermal layer. Hyaluronidase digestion of the cocoon slightly changed the Alcian Blue colour, suggesting the presence of a large amount of acid mucin similar to that found in the skin mucous gland. The results of this study present data for the redistribution of AMPS, which may help in reducing water loss across the cocoon and reabsorption of water in the kidney during aestivation.

The Systemic Administration of Gamma Interferon Inhibits Collagen Synthesis and Acute Inflammation in a Murine Skin Wounding Model

The ability of gamma interferon (IFN-gamma) to affect cutaneous collagen synthesis in vivo was examined in a murine wounding model. Reproducible areas of full-thickness skin necrosis were produced by argon laser radiation. Mice received recombinant murine IFN-gamma (rMuIFN-gamma) (8.7 X 10(3) units/hr) over 14 d via osmotic pumps implanted subcutaneously or intraperitoneally. At 14 and 21 d after wounding, there was less fibrous tissue in healing scars of treated animals as determined by light and transmission electron microscopy. Associated with the decrease in connective tissue was an increase in the acid mucopolysaccharide content of healing scars, which was largely hyaluronate. Quantitative image analysis of electron micrographs confirmed that less collagen was present in healing scars of animals receiving rMuIFN-gamma. The mean cross-sectional area of collagen fibers was smaller in specimens from treated mice, but no difference was seen in the size of collagen fibrils. The time required to obtain full skin closure was also delayed 23%-27% in treated animals. Using this injury model, we also found that rMuIFN-gamma significantly reduced the degree of perilesional erythema surrounding the laser injury sites and, in the first 6 d after wounding, the degree of polymorphonuclear infiltrate present histologically at lesional sites. Indeed, rMuIFN-gamma also decreased the cutaneous accumulation of neutrophils induced by known proinflammatory mediators, such as interleukin 1 and activated serum. Thus, systemically administered IFN-gamma not only down-regulates collagen synthesis in the skin but also modulates in a previously unrecognized manner: neutrophil accumulation at sites of tissue injury in vivo.

On the occurrence of acid mucopolysaccharides in the spermatophores of two marine prawns, Penaeus indiens (Milne-Edwards) and Metapenaeus monoceros (Fabricius) (Crustacea: Macrura)

In marine prawns, spermatophores aid in sperm transfer. The prawn spermatophores, in general, consist of two parts, sperm sac and wing. The former encloses spermatozoa and the latter serves as an adhesive pad during transfer to the female's thelycum. This paper reports on the occurrence of acid mucopolysaccharides (AMPS) in the spermatophores of two penaeid prawns, Penaeus indicus (Milne-Edwards) and Metapenaeus monoceros (Fabricius). They, being the principal component of the spermatophores, have been characterized and quantified in the wing, sperm mass, sperm sac, and crystalline structures. In both prawns, the spermatophore is a simple structure. In P. indicus, it consists of sperm mass, enclosed within sperm sac, to which is attached the foliacious wing. In M. monoceros, the freshly extruded spermatophore consists of sperm mass and milky-white crystalline structures. Isolation and purification of AMPS resulted in the resolution of two toluidine blue-positive AMPS fractions in agarose-gel electrophoresis. Densitometric study of the standard and sample AMPS fractions reveals further that the two fractions correspond to chondroitin sulfate and hyaluronic acid. Quantitative assay on total AMPS shows that the sperm sac of P. indicus contains the maximum amount of AMPS (195.50 μg/mg), whereas its wing contains the least quantity of AMPS (43.68 μg/mg). The qualitative and quantitative variations found in the AMPS content of spermatophoric components of two prawns have been discussed in relation to their adhesion to thelycum as well as sperm storage and maintenance pending fertilization.

Gallbladder adenocarcinoma: prognostic significance of tumor acid mucopolysaccharide content.

The gall bladder mucosa is composed of neutral mucopolysaccharide and protein radical containing secretory cells, protein radical containing migratory cells, and neutral and acid mucopolysaccharides plus sialic acid containing goblet cells. The prognostic significance of histologic or histochemical parameters in gall bladder adenocarcinoma (GBA) are unknown. To determine if histochemical acid mucopolysaccharide content in GBA has prognostic value, GBA histologic sections from 26 advanced stage disease patients participating in Eastern Cooperative Oncology Group (ECOG) treatment study EST-2273 were stained with alcian blue at pH. 1.0, assessed by a pathology panel for either high (greater than 50%) or low (less than 50%) acid mucopolysaccharide content, and correlated with patient survival. Initial panel unanimous concurrence rate on acid mucopolysaccharide content was 88.9%. Median survival times from the start of chemotherapy to date of death for high acid mucopolysaccharide content GBA was 14 weeks versus five weeks for the low content GBA (P less than 0.0001). The results indicate that high acid mucopolysaccharide content in GBA significantly improves prognosis. ECOG recommends stratification by acid mucopolysaccharide content in future GBA treatment investigations.

A HISTOCHEMICAL AND ULTRASTRUCTURAL STUDY OF HUMAN URETHRAL UROEPITHELIUM

The ultrastructure and histochemistry of the human urethral mucosa was studied. By scanning electronmicroscopy (SEM) the cover cell of the urethra was found to be polygonal and with a surface topography characterized by numerous microvilli and micro-ridges. By transmission electronmicroscopy (TEM) the cover cells were shown to be interconnected with tight junctions but to lack the asymmetric luminal membrane and the fusiform vacuoles that characterize urothelium above the bladder neck. Histochemical analyses showed the human urethral cells to harbour large amounts of glycogen, and the glycocalyx facing the urethral lumen displayed high affinity for alcian blue and colloidal iron, indicating the presence of acid mucopolysaccharides. The reactivity with alcian blue appeared at pH 2.5, but was abolished by pre-treatment with sialidase. Studies with TEM or using SEM with energy-dispersive X-ray spectrometry (EDX) confirmed the high content of acid mucopolysaccharides in the luminal glycocalyx of the cover cells by demonstrating high binding capacity for ruthenium red. The quantitative binding of ruthenium red was not influenced by pH shifts between 4.5 and 7.5. Utilizing the SEM + EDX technique, small variations in quantity of negative charge (i.e. of bound ruthenium red) were detected within individual cover cells, but considerable variations were found between cells. The significance of these physicochemical properties of the human urethral lining is discussed with special reference to the previously demonstrated liability of this mucosal surface to interact with microorganisms such as Escherichia coli, Staphylococcus saprophyticus and Neisseria gonorrhoeae.

Studies on skin and urinary mucopolysaccharides in malnourished/vitamin A deficient children

In protein-calorie malnourished children, with or without associated vitamin A deficiency, skin content of acid mucopolysaccharides (MPS) and urinary excretion of MPS and amino sugars were studied. MPS content of skin in both malnourished groups was increased 3–6-fold. This increase was essentially in the non-sulphated component. In normal skin, non-sulphated MPS accounted for 68% of the MPS content, whereas in the malnourished group with vitamin A deficiency it constituted 93%. Urinary excretion of MPS (24 h) was significantly reduced by 50–70% in malnourished groups. This returned to normal levels in the malnourished/vitamin A deficient group when vitamin A injections were administered. Excretion of amino sugars (24 h) in the malnourished groups was also decreased by 50–70%. In normal children 55% of the total amino sugars was dialysable whereas in the malnourished it was increased to 60%. The excretion of protein-bound and dialysable amino sugars was increased to normal levels only in the group given supplements of vitamin A in addition to protein and calories.

Long-term effects of chondrocyte death on rabbit articular cartilage in vivo

The effects of freezing localized areas of rabbit articular cartilage in vivo were studied at two to twelve months morphologically (gross and microscopic examination, including scanning electron microscopy), histochemically (toluidine blue and safranin O), and metabolically (35S uptake). Up to six months the frozen cartilage was intact but appeared to be dead, as shown by the absence of stainable chondrocytes, severely decreased acid mucopolysaccharide content, and absence of significant uptake of 35S. At twelve months fibrillation and softening were evident, clusters of new chondrocytes with surrounding acid mucopolysaccharides were visible microscopically, and scanning electron microscopy revealed an irregular pattern of collagen fibers which were larger than normal, of varying thickness, and broken in some areas. These changes resembled those seen in degenerative joint disease.

MORPHOLOGICAL CHANGES IN THE ARTICULAR CARTILAGE AFTER MENISCECTOMY

The medial meniscus was resected from the right knees of twelve young grivet monkeys that were killed at intervals of twenty-one to 252 days after operation. The knees operated upon and the control knees were investigated radiologically and histologically. Degenerative changes occurred in the medial femoral and tibial condyles. At first there was loss of cells from the superficial layer of the articular cartilage, with a marked decrease in the acid mucopolysaccharide content of the matrix. The chondrocytes in the deeper layer of the non-calcified zone proliferated to form clones before finally degenerating. The acellular cartilage showed splitting, and with progress of the degenerative process there was thinning and erosion of the cartilage. Eventually there was complete loss of articular cartilage with thickening and exposure of the subchondral bone. These degenerative changes were confined to a small area of the articular cartilage and had occurred despite regeneration of the meniscus. The rest of the cartilage looked normal. It is concluded that articular cartilage deprived of the protection of a meniscus may undergo arthritic changes.

Arteriosclerotic Changes within the Diabetic Oral Vasculature

Microscopic sections of diabetic, arteriosclerotic nondiabetic, and nondiabetic nonarteriosclerotic human palate were qualitatively evaluated for small artery colloidal-iron-positive, acid mucopolysaccharide content. The association of abnormal colloidal-iron-positive material with structural detail in the arterial wall was clarified by comparison with specimens of amputated diabetic human legs.

Nicotine effects on the acid mucopolysaccharide content of chick embryo cardiac jelly.

Histochemical studies were made on the developing chick embryo heart to determine the effects of nicotine on acid mucopolysaccharide content during the critical stages of the cardiac morphogenesis. Two-day old embryos were injected with the dosages of nicotine ranging from 1.5 to 3 mg per embryo. The embryos were studied on the 3rd and 4th days of incubation. For an evaluation of acid mucopolysaccharides, the staining procedures of Mowry (1958) and Saunders (1964) were employed. It was found that nicotine decreases the content of acid mucopolysaccharide in the developing cardiac jelly. In the control embryos, the cardiac tissues picked up very intense coloration. At lower dosages (Group A; 1.5 mg/egg), the staining response of the treated heart tissues containing acid mucopolysaccharides was moderate as compared to the control embryos. At higher dosages (Group B; 3 mg/egg), the cardiac tissues were faintly colored as compared to the embryos of Group A. It is possible that the cardiac lesions previously reported in the chick embryos following the administration of nicotine (Gilani, 1971) are due to the reduction of the amount of acid mucopolysaccharides in the developing cardiac jelly--a susceptible period of the genesis of heart.

The development of skin mucous glands ofXenopus laevis during metamorphosis.

As the number of multicellular mucous glands increases in the skin ofXenopus laevis larvae during metamorphosis, the acid mucopolysaccharide (AMPS) content of the skin increases also. Purification of this skin AMPS by polyacrylamide gel electrophoresis revealed a major band characteristic of adult mucous secretion. Incorporation of14C-galactose into this band increases during metamorphosis, but there is still a substantial synthesis of this material in premetamorphic skin that lacks mature mucous glands. This raises the question of whether the epidermal cells that had been making mucus in premetamorphic skin stop this activity as mucous glands develop, or whether they are discarded and replaced by a new population of epidermal cells that do not make mucus unless they develop into a mucous gland.

The influence of age on the wound healing of experimental myocardial infarction in rats: (Autoradiographic studies)

In 6-weeks-old, 5-months-old and 21-months-old rats myocardial infarction was induced by coronary artery ligature. After performing the ligature the animals were administered 3H-thymidine, 3H-proline or 35S-sulphate at different times. The following parameters were determined: number of DNA- and tropocollagen-synthesizing connective tissue cells at the infarction border and at the infarction site; mean silver grain density above the nuclei or cells; duration of a cycle; number of mitoses, and incorporation of radioactive sulphate at the infarction site. In addition, the labelling and mitotic indices as well as the percentage of the standard deviation from the mean values were estimated. The following results were obtained: 1. The rate of granulation tissue formation in the necrotic zone is determined by the mitotic activity of the cells. With advancing age the cell cycles are being prolonged which results in retardation of wound healing. 2. The disturbed DNA-replication in old age is not associated with a time shift in the occurrence of the mitotic and labelling peaks. 3. With advancing age the number of fibroblasts synthesizing collagen precursors decreases. There exists no age-dependence of the 3H-proline incorporation rate, of the intracellular transport, of the synthesis of collagen precursor and of the release of labelled tropocollagen. In all age-groups under study these processes last approximately 4 hours. 4. Collagen fibre formation is accompanied by an increased synthesis of acid mucopolysaccharides. In infarction callosities the content of acid mucopolysaccharides mostly is constant. 5. The proliferating endothelial cells have a pronounced metabolic activity and a markedly short generation time.

Biochemical, histochemical, and autoradiographic evidence for the existence and formation of acid mucopolysaccharides in larvae of Phormia regina

Phormia regina larvae cultures were grown and collected at all stages of their development from hatching to pre-pupation. Preparations from these larvae were analysed for their content of acid mucopolysaccharides (MPS). Numbers of MPS were detected in these preparations and were found to elute in chromatographic patterns that were similar to those shown for authentic MPS standards. Although the MPS appeared to be present at all stages of larval development, the first three days post-hatch were observed to be the most active period for MPS synthesis. Using histochemical and autoradiographic techniques these MPS were found to be present in a variety of organ and tissue sections.

Changes in the content of human aortic glycoproteins and acid mucopolysaccharides in atherosclerosis.

Abstract The concentration of glycoproteins (GP) and acid mucopolysaccharides (AMPS) in apparently normal and atherosclerotic regions of human aortae taken at autopsy was determined both in terms of intimal surface area and dry, defatted, ash-free residue weight (DDAF). Variability due to age was excluded by choosing samples within the same age group. Total GP concentration increased significantly in atherosclerotic tissue as evidenced by increase in total neutral sugars, hexosamine and sialic acid both in terms of surface area and DDAF. The total uronic acid content was not altered. However, the ratio of sulfated:non-sulfated AMPS was significantly higher in atherosclerotic regions indicating that sulfated AMPS concentration is increased in atherosclerotic plaques at the expense of hyaluronic acid. These observations suggest that increased GP-concentration may have a functional role in the formation of atherosclerotic plaque.

Acid mucopolysaccharides (glycosaminoglycans) in normal human kidneys and in kidneys of patients with mucopolysaccharidoses

The acid mucopolysaccharide content, composition and molecular weight distribution in cortex, medulla and papilla of kidneys from normal subjects and from one patient each with mucopolysaccharidosis type I (Hurler) and type II (Hunter) were determined. It was found that heparitin sulfate and hyaluronic acid were the major acid mucopolysaccharides in normal human kidneys, the former predominating in the cortex and the latter in the medulla and papilla. The total amount of acid mucopolysaccharides in the kidneys from the patients with mucopolysaccharidoses was more than ten times higher than in the unaffected controls. In addition, there was an increase of 15- to 50-fold in the dermatan sulfate, and of 3- to 5-fold in the heparin sulfate content and a drastic shift in the molecular weight distribution towards smaller molecules.

組織透過性に対する諸種薬物の影響(第6報)

The mechanism of inflamed dermal tissue permebility in filter paper-implanted rats and the inhibitory effect of anti-inflammatory drugs were investigated. In order to elucidate the mechanism, quantitative changes of connective tissue components [acid mucopolysaccharide (AMPS), glycoprotein (GP) and collegen (C)] in inflamed tissues and the effect of anti-inflammatory drugs were tested. 1) When inflamed tissue was divided into skin side upper fiilter paper and the under muscle one, AMPS content in the muscle side and the GP in both tissues increased very rapidly and markedly. Peaks of both component levels were reached at day 5 or 7. Later AMPS level decreased slightly and thereafter remained at a constant level (almost twice the normal level) until day 25. On the other hand, the GP level thereafter followed a slow reduction, which was more pronounced on the skin side than in muscle one, the level on the skin side fallen to the normal level by day 25. C content in the muscle side of filter paper-implanted rats increased progressively until day 25, while the level on the skin side became markedly lower than normal. 2) The effect of anti-inflammatory drugs (acetylsalicylic acid, aminopyrine, phenylbutazone, bucolome, flufenamic acid, benzydamine HCI, indomethacin, prednisolone and dexamethasone) administered orally once a day for 3 days was examined 3 days after implantation. The increase of AMPS content on the muscle side was inhibited significantly by all these drugs. Acetylsacylic acid, phenylbutazone, flufenamic acid indomethacin and prednisolone also inhibited significantly the increase of GP content in both tissues, whereas benzydamine HCl and dexamethasone effected only in the skin side and bucolome only in muscle side. The decrease of C content in skin side was inhibited remarkably by predni-solone, dexamethasone and indomethacin. 3) The inflamed dermal tissue permeability and GP content in the same region, following a similar pattern, changed during an inflammatory process. A correlation was proved between the inhibitory effect of anti-inflammatory drugs on this two parameters.

Age-related change in acid mucopolysaccharides level and water content in the papillary region of human kidneys.

An attempt has been made to explore the interrelationship between the age-related changes in acid mucopolysaccharides (AMPS) and water content of the renal papilla. Water content of human renal papilla changes significantly with age but the correlation between AMPS and water content of this region is just below the 5% level. On the other hand, the ratio of water content to AMPS decreases significantly with advancing age. Since the change in the ratio of these two substances is interpreted to express mainly the alteration of the ground substance with age, the possibility is discussed that the age-related changes in AMPS and water content of the renal papilla might be related to the decrement in concentrating ability of urine in the elderly person.

The hexosamine and uronic acid contents of the matrix of shells of eggs from pullets fed on diets of different manganese content

Synopsis Three experiments are reported concerning the effect of the manganese content of the diet on the acid mucopolysaccharide content of egg shell matrix. In the first experiment hexosamine was taken as an index of acid mucopolysaccharide, but in experiments 2 and 3 uronic acid was determined instead. It was found that a low‐Mn diet depressed the hexosamine and uronic acid content of the matrix of eggs sampled at the start of lay. However, eggs from low‐Mn fed pullets after three months of lay showed a significant increase in matrix uronic acid. Other characteristics of shells from low‐Mn fed pullets taken at the start of lay were a low weight of shell per unit area and a high weight of matrix per unit area, which led to an abnormally high proportion of matrix to mineral in the shell. As a model for normal shell development eggs were extracted from the uterus at an early and late stage of shell formation. Although these uterine shells resembled the poor shells from the low‐Mn diet from the point of vi...