In vitro synthesis of glycosaminoglycans in endocrine ophthalmopathy

1992 ◽  
Vol 127 (5) ◽  
pp. 397-402 ◽  
Author(s):  
G Kahaly ◽  
C Stover ◽  
J Beyer ◽  
E Otto
Keyword(s):  
Hyaluronic Acid ◽  
Acid Concentration ◽  
Cetylpyridinium Chloride ◽  
Acid Synthesis ◽  
Cell Mediated Immunity ◽  
Glycosaminoglycan Synthesis ◽  
In Vitro Synthesis ◽  
Significant Difference ◽  
Endocrine Ophthalmopathy

The effects of humoral and cell-mediated immunity on the glycosaminoglycan synthesis of retrobulbar fibroblasts was evaluated in patients with endocrine ophthalmopathy. After incubation with IgG and sera, secreted glycosaminoglycans, radiolabeled with D-6-3H-glucosamine and 35sulfate, were precipitated with cetylpyridinium chloride and ethanol. Hyaluronic acid synthesis of human retrobulbar fibroblasts after incubation with sera and IgG and after co-culture with lymphocytes was assessed by means of a radiometric test. Patients' IgG, compared to controls', accounted for a higher secretory stimulation of porcine retrobulbar fibroblasts (as measured by cetylpyridinium chloride precipitation) after 24 and 48 h. Contrasting with 24 h incubation time, glycosaminoglycan values after 48 h were increased two to threefold. Patients' and controls' sera caused earlier and stronger, yet indistinguishable glycosaminoglycan production. Non-sulfated hyaluronic acid was the preponderant glycosaminoglycan secreted into the media by retrobulbar fibroblasts. As assessed with the radiometric test, incubation with patients' and controls' sera and IgG did not reveal a significant difference in stimulating the hyaluronic synthesis of patients' and controls' retrobulbar fibroblasts. When measuring the hyaluronic acid synthesis of controls' and patients' retrobulbar fibroblasts after co-cultivation of lymphocytes, however, patients' lymphocytes had a marked ability to increase the hyaluronic acid concentration compared to controls' lymphocytes. The hyaluronic acid concentration after incubation of a patient's retrobulbar fibroblasts with autologous lymphocytes was markedly more elevated than the intrinsic hyaluronic acid production of retrobulbar fibroblasts. In conclusion, though a significant in vitro influence of patients' IgG and sera on the glycosaminoglycan release of both porcine and human (patients' as well as controls') retrobulbar fibroblasts could not be observed in this study, the indications of a marked stimulatory influence of lymphocytes on the hyaluronic acid secretion of retrobulbar fibroblasts demand further investigation.

scholarly journals The biosynthesis in vitro of chondroitin sulphate in neonatal rat epiphysial cartilage

1972 ◽  
Vol 126 (2) ◽  
pp. 417-432 ◽  
Author(s):  
C. J. Handley ◽  
C. F. Phelps
Keyword(s):  
Hyaluronic Acid ◽  
Chondroitin Sulphate ◽  
Cetylpyridinium Chloride ◽  
Neonatal Rat ◽  
Turnover Rates ◽  
Glycosaminoglycan Synthesis ◽  
Epiphysial Cartilage ◽  
Sulphated Glycosaminoglycans ◽  
Tissue Contents

1. A system is described, which was used to incubate neonatal rat epiphysial cartilage in vitro with [U-14C]glucose and [35S]sulphate. 2. The acid glycosaminoglycans of neonatal rat epiphyses were extracted and fractionated on cetylpyridinium chloride–cellulose columns. The major components were chondroitin 4-sulphate (65%), chondroitin 6-sulphate (15%), hyaluronic acid (4%) and keratan sulphate (2%). 3. The acid-soluble nucleotides and intermediates of glycosaminoglycan synthesis were separated on a Dowex 1 (formate) system. The tissue contents and cellular concentrations of these metabolites were determined. 4. The rates of synthesis of UDP-glucuronic acid and UDP-N-acetyl-hexosamine from [U-14C]glucose were found to be 0.79±0.16 and 3.2±0.08nmol/min per g wet wt. respectively. 5. The incorporation of [U-14C]glucose into the uronic acid and hexosamine moieties of the polymers was also measured and the turnover rates of the glycosaminoglycans were calculated. It was found that chondroitin sulphate was turning over in about 70h and hyaluronic acid in about 120h. 6. The relative rates of synthesis of the sulphated glycosaminoglycans were calculated from [35S]sulphate incorporation and were found to be in good agreement with those obtained from [U-14C]glucose labelling.

Somite chondrogenesis in vitro: 2. Changes in the hyaluronic acid synthesis

1986 ◽  
Vol 18 (2) ◽  
pp. 91-99 ◽  
Author(s):  
Nagaswamisri Vasan ◽  
Karen M. Lamb ◽  
Odette la Manna
Keyword(s):  
Hyaluronic Acid ◽  
Acid Synthesis

scholarly journals Optimization of enzyme cascades for the in vitro synthesis of hyaluronic acid

2018 ◽  
Vol 90 (9) ◽  
pp. 1291-1291
Author(s):  
J. Gottschalk ◽  
A. Eisele ◽  
L. Elling
Keyword(s):  
Hyaluronic Acid ◽  
In Vitro Synthesis ◽  
Enzyme Cascades

scholarly journals Murine SPAM1 is secreted by the estrous uterus and oviduct in a form that can bind to sperm during capacitation: acquisition enhances hyaluronic acid-binding ability and cumulus dispersal efficiency

Reproduction ◽  
2008 ◽  
Vol 135 (3) ◽  
pp. 293-301 ◽  
Author(s):  
Genevieve S Griffiths ◽  
Kimberly A Miller ◽  
Deni S Galileo ◽  
Patricia A Martin-DeLeon
Keyword(s):  
Hyaluronic Acid ◽  
Reproductive Tract ◽  
Immune Serum ◽  
Female Reproductive Tract ◽  
Binding Ability ◽  
Glycosyl Phosphatidylinositol ◽  
Significant Difference ◽  
Surface Remodeling ◽  
Hyaluronic Acid Binding

Sperm uptake of epididymal sperm adhesion molecule 1 (SPAM1)in vitrohas recently been shown to be a marker of sperm maturation, since acquisition of this surface hyaluronidase increases cumulus dispersal efficiency. Here, we demonstrate that this glycosyl phosphatidylinositol-linked sperm antigen, previously shown to be expressed during estrous in the female reproductive tract, is secreted in the uterine and oviductal fluids (ULF and OF respectively) in a 67 kDa form, which can bind to sperm. We show that it can be acquired by caudal sperm fromSpam1null,Spam1-deficient mutant, and wild-type (WT) micein vitroduring incubation in ULF or OF at 37 °C, as detected by immunocytochemistry and flow cytometry. SPAM1 binding after ULF incubation was localized predominantly to the acrosome and the mid-piece of the flagella ofSpam1null sperm in a pattern identical to that of WT sperm. After ULF incubation, WT sperm demonstrated a significantly (P<0.001) enhanced hyaluronic acid-binding ability, and the involvement of SPAM1 in this activity was shown by a significant (P<0.001) decrease in binding when sperm were exposed to SPAM1 antiserum-inhibited ULF. Importantly, whenSpam1null sperm were exposed to ULF with SPAM1 accessible (in the presence of pre-immune serum) or inaccessible (in the presence of SPAM1 antiserum) for uptake, there was a significant difference in cumulus dispersal efficiency. Taken together, these results suggest that in the sperm surface remodeling that occurs prior to and during capacitation, the fertilizing competence of sperm is increased via acquisition of SPAM1, and likely other hyaluronidases, from the female tract.

scholarly journals The effect of mouthwashes on fluoride dentifrices in preventing dental abrasion or erosion

2021 ◽  
Vol 14 (3) ◽  
pp. 361-366
Author(s):  
Rachna Raj ◽  
◽  
Safia Haideri ◽  
Bipin Kumar Yadav ◽  
Joohi Chandra ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Cetylpyridinium Chloride ◽  
Tooth Wear ◽  
The Other ◽  
Surface Loss ◽  
Significant Difference ◽  
Mineralized Tissues ◽  
Ph Cycling ◽  
The Impact

Erosive tooth wear (ETW) refers to the chemical dissolution of mineralized tissues by acids of non-bacterial origin. It occurs in the primary as well as the permanent dentition. In this study, our objectives were to investigate and compare the impact of chlorhexidine gluconate (CHX), essential oils (EO), and cetylpyridinium chloride (CPC) on ETW protection produced by conventional fluoride kinds of toothpaste. A clinically relevant in-vitro erosion/abrasion pH cycling model was employed to test the effect of the aforementioned mouthwashes on modulating the ability of NaF and SnF2 types of toothpaste. The mean dentin surface loss associated with NaF toothpaste was significantly lower than for the SnF2 toothpaste. On the other hand, enamel surface loss with SnF2 toothpaste was significantly lower than for the NaF toothpaste. Also, the surface loss of erosion was significantly higher when associated with abrasion than without brushing and for both enamel and dentin. There was no significant difference in the surface loss among all types of mouthwash. Commonly used types of mouthwash containing antimicrobial agents or additional fluoride do not impact fluoride toothpaste action on erosion/abrasion. Also, considering erosion only, the tested SnF2 dentifrice provided better protection against surface loss of enamel than the other.

scholarly journals Spraying of dsRNA Molecules Derived from Phytophthora Infestans, as a Plant Protection Strategies for the Management of Potato Late Blight

2021 ◽  
Author(s):  
Sundaresha S. ◽  
Sanjeev Sharma ◽  
Aarti Bairwa ◽  
Maharishi Tomar ◽  
Ravinder Kumar ◽  
...  
Keyword(s):  
Late Blight ◽  
Phytophthora Infestans ◽  
Disease Severity ◽  
Plant Protection ◽  
Early Stage ◽  
Microscopy Imaging ◽  
In Vitro Synthesis ◽  
Significant Difference ◽  
Nano Clay

Phytophthora. infestans is a well known late blight causing oomycetes pathogen. It evolves and adapts to the host background and new fungicide molecules rapidly within a few years of their release, may be due to the predominance of transposable elements in its genome. Frequent and huge applications of fungicides cause environmental concerns. Here we developed target specific RNA interference based molecules, that are capable of effectively reducing the late blight infection. cDNA microarray expression data was used for the selection of genes involved in the early stage of infection process, sporulation etc. The in vitro synthesis of double stranded RNA molecule, targeting SDH, EF-1&alpha;, GPI-HAM344, PLD-3 and HSP-90 encoding genes revealed the reduction in growth, sporulation and symptom expression, which were subsequently assessed by culture bioassay, detached leaf assay and topical application methods. The multiple genes targeted dsRNA nano clay sprayed plants showed enhanced disease resistance (4% disease severity) and least sporulation (&lt;1x103), compared to naked dsRNA spray. Use of nano clay was assumed to be involved in the effective delivery, protection and boosting the action of RNAi in potato plants. A significant difference in the growth, sporulation count, disease severity and reduced expression of the genes and confocal microscopy imaging authenticates the effects of SIGS on late blight disease progression. Our research demonstrated that topical dsRNA nano clay spray under the open-air environment could be an alternative to chemical fungicides and transgenic approaches as a novel plant protection strategy for late blight in an environmentally friendly manner.

scholarly journals Key Factors for A One-Pot Enzyme Cascade Synthesis of High Molecular Weight Hyaluronic Acid

2019 ◽  
Vol 20 (22) ◽  
pp. 5664 ◽  
Author(s):  
Gottschalk ◽  
Zaun ◽  
Eisele ◽  
Kuballa ◽  
Elling
Keyword(s):  
Molecular Weight ◽  
Hyaluronic Acid ◽  
Pasteurella Multocida ◽  
Ph Value ◽  
Key Factors ◽  
One Pot ◽  
Data Set ◽  
In Vitro Synthesis ◽  
Enzyme Cascade

In the last decades, interest in medical or cosmetic applications of hyaluronic acid (HA) has increased. Size and dispersity are key characteristics of biological function. In contrast to extraction from animal tissue or bacterial fermentation, enzymatic in vitro synthesis is the choice to produce defined HA. Here we present a one-pot enzyme cascade with six enzymes for the synthesis of HA from the cheap monosaccharides glucuronic acid (GlcA) and N-acetylglucosamine (GlcNAc). The combination of two enzyme modules, providing the precursors UDP–GlcA and UDP–GlcNAc, respectively, with hyaluronan synthase from Pasteurella multocida (PmHAS), was optimized to meet the kinetic requirements of PmHAS for high HA productivity and molecular weight. The Mg2+ concentration and the pH value were found as key factors. The HA product can be tailored by different conditions: 25 mM Mg2+ and 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES)-NaOH pH 8 result into an HA product with high Mw HA (1.55 MDa) and low dispersity (1.05). Whereas with 15 mM Mg2+ and HEPES–NaOH pH 8.5, we reached the highest HA concentration (2.7 g/L) with a yield of 86.3%. Our comprehensive data set lays the basis for larger scale enzymatic HA synthesis.

scholarly journals Competitive Inhibition of Mammalian Hyaluronidase by Tomato Saponin, Esculeoside A

2018 ◽  
Vol 13 (11) ◽  
pp. 1934578X1801301 ◽  
Author(s):  
Jian-Rong Zhou ◽  
Souta Kimura ◽  
Toshihiro Nohara ◽  
Kazumi Yokomizo
Keyword(s):  
Hyaluronic Acid ◽  
Kinetic Analysis ◽  
Competitive Inhibition ◽  
Tomato Fruits ◽  
Hyaluronidase Activity ◽  
Significant Difference ◽  
Esculeoside A ◽  
Menten Constant

Esculeoside A, a glycoside of spirosolane-type, is identified as a major component in ripe tomato fruits. Our previous study showed that esculeoside A inhibited hyaluronidase activity in vitro and ameliorated experimental dermatitis in vivo. The aim of this present study is to investigate the inhibition mode on mammalian hyaluronidase by esculeoside A. Measured by a modified Morgan-Elson method, the present kinetic analysis of the hydrolysis reaction using hyaluronic acid revealed that no significant difference was observed in velocity maximum Vmax, and Michaelis-Menten constant Km was shown as 0.74 mM in the absence of esculeoside A, was increased as 1.32 and 1.98 mM with 3 and 30 μM of esculeoside A, respectively. Thus suggested that the inhibition mode on hyaluronidase by esculeoside A was competitive. This competitive inhibition on hyaluronidase activity may become valuable in the amelioration of mice experimental dermatitis by esculeoside A.

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