Hyaluronan and hyaluronidases are critical in tissue remodeling, inflammation, and tumor progression. This chapter provides a comprehensive guide to hyaluronan zymography, a powerful technique for detecting and quantifying hyaluronidase activity in complex biological samples. The method involves separating proteins by polyacrylamide gel electrophoresis with hyaluronan incorporated into the gel matrix. Following electrophoresis, the gel is incubated to allow hyaluronidases to degrade the hyaluronan substrate, resulting in clear digestion zones. Detailed protocols for sample preparation and the zymographic process are included, offering researchers a robust tool for studying hyaluronidase activity and regulation in various biological contexts.
An increasing number of studies have characterized the bone as an endocrine organ, and that bone secreted factors may not only regulate local bone remodeling, but also other tissues and whole-body metabolic functions. The precise nature of these regulatory factors and their roles at bridging the bone, bone marrow adipose tissue, extramedullary body fat and whole-body energy homeostasis are being explored. In this study, we report that KIAA1199, a secreted factor produced from bone and bone marrow, previously described as an inhibitor of bone formation, also plays a role at promoting adipogenesis. KIAA1199-deficient mice exhibit reduced bone marrow adipose tissue, subcutaneous and visceral fat tissue mass, blood cholesterol, triglycerides, free fatty acids and glycerol, as well as improved insulin sensitivity in skeletal muscle, liver and fat. Moreover, these mice are protected from the detrimental effects of high-fat diet feeding, with decreased obesity, lower blood glucose and glucose tolerance, as well as decreased adipose tissue inflammation, insulin resistance and hepatic steatosis. In human studies, plasma levels of KIAA1199 or its expression levels in adipose tissue are positively correlated with insulin resistance and blood levels of cholesterol, triglycerides, free fatty acids, glycerol, fasting glucose and HOMA-IR. Mechanistically, KIAA1199 mediates its effects on adipogenesis through modulating osteopontin-integrin and AKT / ERK signaling. These findings provide evidence for the role of bone secreted factors on coupling bone, fat and whole-body energy homeostasis.
- MeSH
- adipogeneze * fyziologie MeSH
- dieta s vysokým obsahem tuků MeSH
- energetický metabolismus * MeSH
- hyaluronoglukosaminidasa MeSH
- inzulinová rezistence MeSH
- lidé MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- obezita metabolismus MeSH
- proteiny * metabolismus MeSH
- tuková tkáň metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BackgroundSulodexide is a glycosaminoglycan-based drug prescribed to patients with angiopathy. We performed a pilot study to investigate whether sulodexide positively modulates the endothelial glycocalyx (EG) layer and the microcirculation in a porcine model of EG enzymatic damage. The EG is a sugar-based endothelial lining that is involved in the physiology of the capillary wall and the pathogenesis of many diseases.MethodsEG damage was induced in eight piglets by hyaluronidase III and heparanase I given intravenously. Four animals received sulodexide 600 IU intravenously before the enzymes and four animals after the enzymes were administered. Four animals constituted a control group. Sublingual microcirculation by side-stream dark field imaging and plasmatic concentration of syndecan-1 by ELISA were measured at baseline, 20 min after intervention, and at the 40th, and 60th minute onwards. The statistics were performed with a one-way ANOVA test with Turkey's correction for multiple comparisons testing. Timepoint comparison was performed by Student t-test or Mann-Whitney test.ResultsAt baseline, there were no statistically significant differences between the animal groups. After the intervention, the levels of syndecan-1 were significantly lower in the control group. While there were no differences between the two intervention groups. The sublingual microcirculation analysis showed that the DeBacker score was significantly higher in the control group. At 60 min, there was also a statistically significant difference in DeBacker score between the groups (8.1 ± 1.6 mm-1 in the group with enzymes given first and 11 ± 0.92 mm-1 in the group with sulodexide given first, p = 0.03). The analysis of the proportion of perused vessels did not show any statistically significant differences.ConclusionThe results of the study demonstrated a working model of EG damage but no specific action of sulodexide on EG modulation. In the sublingual microcirculation analysis, the sulodexide reduced the fall in absolute tissue perfusion in 60 min.
- MeSH
- cévní endotel * účinky léků MeSH
- glykokalyx * účinky léků metabolismus MeSH
- glykosaminoglykany * farmakologie MeSH
- hyaluronoglukosaminidasa MeSH
- mikrocirkulace účinky léků MeSH
- modely nemocí na zvířatech MeSH
- pilotní projekty MeSH
- prasata MeSH
- syndekan-1 krev MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Injekční aplikace výplňového materiálu zejména do oblasti obličeje jsou populární estetickou procedurou, která se využívá k obnovení objemu, zlepšení kontur a vyhlazení vrásek. Za nejbezpečnější je považována kyselina hyaluronová, ale i ta nese riziko vzniku vaskulárních komplikací. Tyto komplikace mohou být závažné a vést k ischemii, nekróze tkání nebo dokonce slepotě či centrální mozkové příhodě. Tento článek se zaměřuje na identifikaci rizik spojených s vaskulárními komplikacemi při aplikaci dermálních výplní a na strategie pro jejich prevenci a řešení.
The application of filler material, particularly in the facial area, is a popular aesthetic procedure used to restore volume, improve contours and smooth wrinkles. Hyaluronic acid is considered the safest option, but it too carries the risk of vascular complications. These complications can be severe and lead to ischemia, tissue necrosis, or even blindness or stroke. This article focuses on identifying the risks associated with vascular complications associated with dermal filler application and strategies for prevention and management.
- MeSH
- arteria centralis retinae patologie MeSH
- dermální výplně škodlivé účinky MeSH
- hyaluronoglukosaminidasa * farmakologie škodlivé účinky MeSH
- injekce subkutánní škodlivé účinky MeSH
- kyselina hyaluronová škodlivé účinky MeSH
- lidé MeSH
- nemoci cév * chemicky indukované patologie prevence a kontrola terapie MeSH
- nežádoucí účinky léčiv patologie prevence a kontrola terapie MeSH
- oční symptomy MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
BACKGROUND: Endothelial glycocalyx (EG) plays a crucial role in maintaining the plasma proteins within the intravascular space. OBJECTIVE: We studied whether exogenous albumin protects the EG in an experimental model of EG enzymatic damage in rats. METHODS: Rats were divided into three groups of 10 animals that received (1) Evans blue (2) Evans blue + hyaluronidase, or (3) Evans blue + hyaluronidase + 20% human albumin via the tail vein. Spectrophotometric analysis was performed 2 h later to quantify the leakage of Evans blue-labeled albumin into the heart, lungs, brain, kidneys, liver, small intestine, spleen, and skeletal muscle. RESULTS: Administration of hyaluronidase numerically increased the capillary leakage of Evans blue in all examined tissues. Co-administration of albumin decreased the leakage of albumin in all tissues except the heart. In the lungs, the ratio between the absorbance and dry organ weight decreased from 5.3 ± 2.4 to 1.7 ± 0.5 (mean ± SD) (P < 0.002), and in the liver, the absorbance decreased from 2.2 ± 0.7 to 1.5 ± 0.4 (P < 0.011). CONCLUSION: Exogenous albumin decreased the capillary leakage of albumin which was interpreted as a sign of maintained EG integrity.
- MeSH
- albuminy * metabolismus MeSH
- cévní endotel účinky léků metabolismus MeSH
- Evansova modř MeSH
- glykokalyx * metabolismus účinky léků MeSH
- hyaluronoglukosaminidasa farmakologie MeSH
- kapilární permeabilita * účinky léků MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- potkani Wistar MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Hyaluronan, a linear glycosaminoglycan comprising D-N-acetylglucosamine and D-glucuronic acid, is the main component of the extracellular matrix. Its influence on cell proliferation, migration, inflammation, signalling, and other functions, depends heavily on its molecular weight and chemical modification. Unsaturated HA oligosaccharides are available in defined length and purity. Their potential therapeutic utility can be further improved by chemical modification, e. g., reduction. No synthesis of such modified oligosaccharides, either stepwise or by hyaluronan cleavage, has been reported yet. Here we show a three-step synthesis (esterification, depolymerization and reduction) of unsaturated even numbered hyaluronan oligosaccharides with carboxylates and the reducing terminus reduced to an alcohol. Particular oligosaccharides were synthesised. The modified oligosaccharides are not cleaved by mammalian or bacterial hyaluronidase and do not affect the growth of mouse and human fibroblasts. Further, MTT and NRU viability tests showed that they inhibit the growth of human colon carcinoma cells HT-29 by 20-50 % in concentrations 500-1000 μg/mL. Interestingly, this effect takes place regardless of CD44 receptor expression and was not observed with unmodified HA oligosaccharides. These compounds could serve as enzymatically stable building blocks for biologically active substances.
- MeSH
- antigeny CD44 metabolismus MeSH
- buňky HT-29 MeSH
- cytostatické látky * farmakologie chemie chemická syntéza MeSH
- fibroblasty účinky léků MeSH
- hyaluronoglukosaminidasa * metabolismus antagonisté a inhibitory MeSH
- kyselina hyaluronová * chemie farmakologie MeSH
- lidé MeSH
- myši MeSH
- oligosacharidy * chemie farmakologie MeSH
- proliferace buněk * účinky léků MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND AND AIMS: ADVANCE-CIDP 1 evaluated facilitated subcutaneous immunoglobulin (fSCIG; human immunoglobulin G 10% with recombinant human hyaluronidase) efficacy and safety in preventing chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) relapse. METHODS: ADVANCE-CIDP 1 was a phase 3, double-blind, placebo-controlled trial conducted at 54 sites in 21 countries. Eligible adults had definite or probable CIDP and adjusted Inflammatory Neuropathy Cause and Treatment (INCAT) disability scores of 0-7 (inclusive), and received stable intravenous immunoglobulin (IVIG) for ≥12 weeks before screening. After stopping IVIG, patients were randomized 1:1 to fSCIG 10% or placebo for 6 months or until relapse/discontinuation. fSCIG 10% was administered at the same dose (or matching placebo volume) and interval as pre-randomization IVIG. The primary outcome was patient proportion experiencing CIDP relapse (≥1-point increase in adjusted INCAT score from pre-subcutaneous treatment baseline) in the modified intention-to-treat population. Secondary outcomes included time to relapse and safety endpoints. RESULTS: Overall, 132 patients (mean age 54.4 years, 56.1% male) received fSCIG 10% (n = 62) or placebo (n = 70). CIDP relapse was reduced with fSCIG 10% versus placebo (n = 6 [9.7%; 95% confidence interval 4.5%, 19.6%] vs n = 22 [31.4%; 21.8%, 43.0%], respectively; absolute difference: -21.8% [-34.5%, -7.9%], p = .0045). Relapse probability was higher with placebo versus fSCIG 10% over time (p = .002). Adverse events (AEs) were more frequent with fSCIG 10% (79.0% of patients) than placebo (57.1%), but severe (1.6% vs 8.6%) and serious AEs (3.2% vs 7.1%) were less common. INTERPRETATION: fSCIG 10% more effectively prevented CIDP relapse than placebo, supporting its potential use as maintenance CIDP treatment.
- MeSH
- chronická zánětlivá demyelinizační polyneuropatie * farmakoterapie MeSH
- dospělí MeSH
- hyaluronoglukosaminidasa terapeutické užití MeSH
- intravenózní imunoglobuliny terapeutické užití MeSH
- lidé středního věku MeSH
- lidé MeSH
- lokální recidiva nádoru chemicky indukované farmakoterapie MeSH
- výsledek terapie MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- práce podpořená grantem MeSH
- randomizované kontrolované studie MeSH
Aging is a complex physiological process that can be accelerated by chemical (high blood glucose levels) or physical (solar exposure) factors. It is accompanied by the accumulation of altered molecules in the human body. The accumulation of oxidatively modified and glycated proteins is associated with inflammation and the progression of chronic diseases (aging). The use of antiglycating agents is one of the recent approaches in the preventive strategy of aging and natural compounds seem to be promising candidates. Our study focused on the anti-aging effect of the flavonoid hesperetin, its glycoside hesperidin and its carbohydrate moieties rutinose and rhamnose on young and physiologically aged normal human dermal fibroblasts (NHDFs). The anti-aging activity of the test compounds was evaluated by measuring matrix metalloproteinases (MMPs) and inflammatory interleukins by ELISA. The modulation of elastase, hyaluronidase, and collagenase activity by the tested substances was evaluated spectrophotometrically by tube tests. Rutinose and rhamnose inhibited the activity of pure elastase, hyaluronidase, and collagenase. Hesperidin and hesperetin inhibited elastase and hyaluronidase activity. In skin aging models, MMP-1 and MMP-2 levels were reduced after application of all tested substances. Collagen I production was increased after the application of rhamnose and rutinose.
- MeSH
- hesperidin * farmakologie MeSH
- hyaluronoglukosaminidasa MeSH
- kolagenasy metabolismus MeSH
- lidé MeSH
- pankreatická elastasa MeSH
- rhamnosa * farmakologie MeSH
- stárnutí kůže * účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- dimethylsulfoxid aplikace a dávkování terapeutické užití MeSH
- hojení ran * MeSH
- hyaluronoglukosaminidasa aplikace a dávkování terapeutické užití MeSH
- lidé MeSH
- nádory * komplikace ošetřování MeSH
- onkologické ošetřovatelství * metody MeSH
- protinádorové látky škodlivé účinky MeSH
- radiodermatitida ošetřování MeSH
- radioterapie škodlivé účinky MeSH
- rány a poranění etiologie ošetřování MeSH
- týmová péče o pacienty MeSH
- Check Tag
- lidé MeSH
Preparáty pro imunoglobulinovou substituční léčbu slouží k náhradě nedostatečné tvorby opsonizačních a neutralizačních protilátek proti široké škále patogenů u pacientů s vrozenými nebo získanými poruchami tvorby protilátek. Mezi základní způsoby aplikace patří intravenózní, konvenční subkutánní a nejnověji také subkutánní podání facilitované hyaluronidázou (fSCIG). Podkožní aplikace tohoto enzymu v úvodu léčby umožní dočasně rozmělnit extracelulární matrix, a tím pádem aplikovat objem normálního lidského imunoglobulinu v dávce a intervalu podání jako při intravenózní aplikaci (0,4-0,8 g/kg/měsíc jednou za 2-4 týdny). Biologická dostupnost fSCIG léčby se pohybuje kolem 93 % a je vyšší než při konvenčním subkutánním podávání. Většinou dochází k aplikaci do 1-2 míst. Maximální doporučená rychlost podávání je 300 ml/h. Mezi nejčastější nežádoucí účinky léčby patří lokální reakce charakteru bolesti či dyskomfortu v místě vpichu, svědění, zarudnutí nebo otok. Specifickým problémem fSCIG léčby je možná tvorba protilátek proti hyaluronidáze, které však nejsou neutralizační a jejich přítomnost nekoreluje s výskytem nežádoucích reakcí. Základní výhodou fSCIG léčby je subkutánní podávání v delším časovém intervalu v domácím prostředí.
The immunoglobulin replacement therapy serves to compensate for the insufficient production of opsonizing and neutralizing antibodies against a wide range of pathogens in patients with congenital or acquired antibody production disorders. The basic routes of administration include intravenous, conventional subcutaneous and subcutaneous administration facilitated by hyaluronidase (fSCIG). Subcutaneous administration of this enzyme at the beginning of the treatment allows to temporarily grind the extracellular matrix and thus use the volume of normal human immunoglobulin in the dose and interval of administration as in intravenous route of administration (0.4-0.8 g/kg/month once every 2-4 weeks). The bioavailability of fSCIG treatment is around 93 % and is higher than with conventional subcutaneous administration. It is usually applied to 1-2 sites. The maximum recommended infusion rate is 300ml/h. The most common side effects of treatment include local reactions of pain or discomfort at the injection site, itchy, redness or swelling. The formation of antibodies against hyaluronidase is possible, however they are not neutralizing and their presence does not correlate with the occurrence of adverse reactions. The main advantage of fSCIG treatment is subcutaneous administration over a longer period of time in the home environment.
- Klíčová slova
- facilitované subkutánní podávání,
- MeSH
- hyaluronoglukosaminidasa aplikace a dávkování terapeutické užití MeSH
- imunoglobuliny * aplikace a dávkování terapeutické užití MeSH
- injekce subkutánní MeSH
- lidé MeSH
- syndromy imunologické nedostatečnosti * farmakoterapie MeSH
- způsoby aplikace léků MeSH
- Check Tag
- lidé MeSH
