Hyaluronic acid (HA), an extracellular biopolymer found throughout the human body, holds promise as a biocompatible and biodegradable scaffold material. High molecular weight (HMW) HA degrades, generating low molecular weight (LMW) fragments with distinct properties. These fragments can influence the behaviour of cells, including human dental pulp stem cells (hDPSCs) incorporated into HA-containing hydrogels or scaffolds. Therefore, a comprehensive examination of the impact of a range of HA molecular weights on hDPSCs is essential before designing HA-based scaffolds for these cells. hDPSC lines were cultured with LMW HA (800 Da, 1600 Da, 15 kDa), medium molecular weight HA (237 kDa), or HMW HA (1500 kDa) over six passages. The various molecular weights had negligible effects on hDPSCs viability, morphology, adhesion, or relative telomere length. Furthermore, the expression of key surface stemness markers (CD29, CD44, CD73, CD90) remained unaltered. HA did not induce osteogenic, chondrogenic, or adipogenic differentiation. Moreover, the potential for chondrogenic and osteogenic differentiation was not adversely affected by LMW or HMW HA. Various molecular weights of HA seem safe, biocompatible and therefore suitable components for hDPSCs-containing scaffolds. These findings affirm that the hDPCSs will not be negatively affected by HA fragments resulting from scaffold degradation.
Introduction: Thalassemia is a healthcare challenging disease all over the world. It imparts a great burden on patients' families and healthcare institutions. Scientists focus on new aspects to overcome these challenges and increase patient tolerance of disease complications. This study aims to quantify β-Hydroxybutyrate Dehydrogenase BHBDH activity in thalassemia patients compared to the control group and their correlation with the patient's demographic characteristics.Methods: To do so, serum was collected from patients and the control group and analyzed biochemically for targeted laboratory tests. We determined β-Hydroxybutyrate Dehydrogenase from normal human serum using biochemical molecular techniques.Results: The results showed that BHBDH activity is significantly higher in the patients compared to the control group regardless of age, sex, or marital status. The results confirmed that enzyme activity and the purification folds were (0.0214 U/ml) and (51.7) respectively for the partially purified enzyme. Furthermore, the proportional molecular weight of the incompletely isolated β-Hydroxybutyrate Dehydrogenase was (125.8±0.5 kDa) using gel filtration chromatography. The comparative molecular weight of the subunit of partially isolated β-Hydroxybutyrate Dehydrogenase was (32.1±0.5 kDa) using SDS-PAGE.Conclusion: we demonstrate that BHBDH enzymatic activity is higher than control and this could be a prognostic or diagnostic tool in thalassemia patients.
- MeSH
- beta-talasemie * krev MeSH
- biochemická analýza krve metody MeSH
- diagnostické techniky molekulární metody MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- gelová chromatografie MeSH
- hydroxybutyrátdehydrogenasa * chemie fyziologie izolace a purifikace krev MeSH
- lidé MeSH
- molekulová hmotnost MeSH
- statistika jako téma MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- klinická studie MeSH
- Geografické názvy
- Irák MeSH
Cyanobacteria produce a wide range of metabolites of interest for industrial or medical use. The cultivation of freshwater Nostoc cf. linckia yielded 5.4 g/L of a crude exopolysaccharide (cEPS) with a molecular weight of 1.31 × 105 g/mol. Ion-exchange chromatography of cEPS yielded two dominant fractions, EPS-1 and EPS-2, differing in molecular weight. The lower molecular weight fraction (EPS-1) was subjected to structural studies. Results of chemical and spectroscopic analyses showed that three of the four dominant sugars, glucose, galactose and xylose are 1,4-linked in the backbone in the following order: [→4)-β-D-Xylp-(1 → 4)-β-D-Glcp-(1 → 4)-α-D-Galp-(1 → 4)-β-D-Glcp-(1→]n. Terminal mannose residues were identified as side chains linked at C3 of every third backbone xylose and every second glucose is branched at C6 by 3-O-lactyl-β-D-glucuronic acid (nosturonic acid). Antioxidant properties of EPS were tested using two in vitro methods. Both assays showed that the cEPS was more active than purified EPS-1 and EPS-2 fractions and deproteinized EPS.
- MeSH
- antioxidancia chemie MeSH
- bakteriální polysacharidy analýza chemie MeSH
- galaktosa chemie MeSH
- glukosa chemie MeSH
- kyselina glukuronová chemie MeSH
- magnetická rezonanční spektroskopie metody MeSH
- molekulární struktura MeSH
- molekulová hmotnost MeSH
- Nostoc chemie MeSH
- xylosa chemie MeSH
- Publikační typ
- časopisecké články MeSH
An exopolysaccharide (EPS) synthesizing potentially probiotic Gram-positive bacterial strain was isolated from fish (Tor putitora) gut, and its EPS was structurally characterized. The isolate, designated as FW2, was identified as Lactobacillus reuteri through 16S rRNA gene sequencing and phylogenetic analysis. This isolate produces fructan-type EPS using sucrose as a substrate. Based on 13C-NMR spectroscopy, methylation analysis and monosaccharide composition, the EPS was identified as a linear levan polymer with fructose as main constituent linked via β(2 → 6) linkages. Based on molecular weight (MW) distribution, two groups of levan were found to be produced by the isolate FW2: one with high MW (4.6 × 106 Da) and the other having much lower MW (1.2 × 104 Da). The isolate yielded about 14 g/L levan under optimized culturing parameters including aeration conditions, pH, temperature and substrate concentration. The obtained bimodal molecular weight linear levan is the first of its type to be synthesized by a L. reuteri isolate from fish gut. Bimodal molecular weight prebiotic levan together with the probiotic potential of the producing strain would provide a new promising synbiotic combination for use in aqua culture.
- MeSH
- fruktany MeSH
- fylogeneze MeSH
- Limosilactobacillus reuteri * genetika MeSH
- molekulová hmotnost MeSH
- RNA ribozomální 16S genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The conditions determining network-forming and aggregation properties of hyaluronan on the mica surface were studied. The hyaluronan was deposited on the surface from aqueous and saline solutions and attached by a bivalent cation. The morphology of the immobilized assemblies was characterized by atomic force microscopy. The experimental results show that the morphology and size of the aggregates as well as the density of the interconnecting fibrillar network, both made of hyaluronan, at the liquid-solid phase interface are determined not only by its molecular weight or concentration in solution, but also by the dissolution conditions and storage time. These findings extend the current state of knowledge about the conformational variability of this biologically important polymer. Understanding the conformational variability is of great importance, as it governs the physiological functions of hyaluronan, as well as its processability and formulations. That in turn determines its usability in different pharmacological and biomaterial applications.
- MeSH
- hydrofobní a hydrofilní interakce MeSH
- hypertonický solný roztok chemie MeSH
- kyselina hyaluronová chemie MeSH
- mikroskopie atomárních sil metody MeSH
- molekulární struktura MeSH
- molekulová hmotnost MeSH
- polymery chemie MeSH
- povrchové vlastnosti MeSH
- rozpustnost MeSH
- silikáty hliníku chemie MeSH
- skladování léků MeSH
- voda chemie MeSH
- vodíková vazba MeSH
- Publikační typ
- časopisecké články MeSH
Heterologously expressed and purified azoreductase enzyme from facultative Klebsiella pneumoniae was used to degrade sulphonated azo dye. Methyl orange (MO) was used as the model dye to study the azo dye decolorization potential of the purified enzyme at different conditions. The enzyme had maximum activity at 40 °C and pH 8.0. The enzyme was observed to be thermo-stable as some enzyme activity was retained even at 80 °C. The apparent kinetic parameters, i.e., appKm and appVmax, for azoreductase using MO as a substrate were found to be 17.18 μM and 0.08/min, respectively. The purified enzyme was able to decolorize approximately 83% of MO (20 μM) within 10 min in the presence of NADH. Thus, efficient decolorization of MO was observed by the purified enzyme. The recombinant enzyme was purified approximately 18-fold with 46% yield at the end of four steps of the purification process. Enzyme was present in a tetrameric structure as confirmed by the volume at which protein was eluted in gel filtration chromatography, and the monomeric molecular mass of enzyme was found to be 23 kDa on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The dye degradation efficiency of azoreductase cloned from Klebsiella pneumoniae and purified from recombinant Escherichia coli was observed to be much higher as compared with the efficiencies of the reported azoreductases from other bacterial strains. In the present study, we report the purification and characterization of the azoreductase cloned from Klebsiella pneumoniae and expressed in Escherichia coli.
- MeSH
- azosloučeniny metabolismus MeSH
- bakteriální proteiny chemie genetika izolace a purifikace metabolismus MeSH
- barvicí látky metabolismus MeSH
- biodegradace MeSH
- Escherichia coli genetika metabolismus MeSH
- kinetika MeSH
- Klebsiella pneumoniae enzymologie genetika MeSH
- koncentrace vodíkových iontů MeSH
- molekulová hmotnost MeSH
- nitroreduktasy chemie genetika izolace a purifikace metabolismus MeSH
- rekombinantní proteiny chemie genetika izolace a purifikace metabolismus MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
This study evaluated the effect of low-molecular weight chitosan on Staphylococcus epidermidis, a common colonizer of joint implants and other prosthetic devices. We have also attempted to elucidate its mechanism of action. Chitosan was found to be effective against both the planktonic and biofilm cells (MIC80 35-40 mg/L; MBIC80 40-150 mg/L), in contrast to the antibiotics erythromycin and tetracycline with no antibiofilm activity (MBIC80 not found). In combination, chitosan had an additive effect with antibiotics on suspension growth of S. epidermidis (FICi 0.7-1.0), and the combinatory action caused a complete inhibition of biofilm metabolic activity in some cases. In addition, chitosan caused rapid cellular damage and enhanced antihaemolytic activity of tetracycline in combination towards S. epidermidis biofilm cells. Chitosan efficiently inhibited S. epidermidis growth acting via cell membrane damage, yet the extent of antimicrobial and antibiofilm activities was quite strain-specific. It was proved to be a very efficient antimicrobial agent worth further examination as a potent candidate in pharmaceutical research. Apart from antimicrobial activity, it also acted as antivirulence enhancing agent which is a very promising strategy for alternative infectious diseases treatment.
The molecular weight (Mw) of dextran derivatives, such as regioselectively oxidized dicarboxydextran (DXA), is greatly influencing their faith in an organism, which could be possibly used to improve anticancer drug delivery. Here we present a modified method of sulfonation-induced chain scission allowing direct and accurate control over the Mw of DXA without increasing its polydispersity. Prepared DXA derivatives (Mw = 10-185 kDa) have been conjugated to cisplatin and the Mw of the carrier found to have a significant impact on cisplatin release rates, in vitro cytotoxicity, and migrastatic potential. Conjugates with the high-Mw DXA showed particularly increased anticancer efficacy. The best conjugate was four times more effective against malignant prostatic cell lines than free cisplatin and significantly inhibited the ovarian cancer cell migration. This was traced to the characteristics of spontaneously formed cisplatin-crosslinked DXA nanogels influenced by Mw of DXA and amount of loaded cisplatin.
- MeSH
- adenokarcinom farmakoterapie metabolismus MeSH
- antitumorózní látky chemie farmakologie MeSH
- buňky A549 MeSH
- cisplatina chemie farmakologie MeSH
- dextrany chemie MeSH
- lidé MeSH
- molekulová hmotnost MeSH
- nádory prostaty farmakoterapie metabolismus MeSH
- nádory vaječníků farmakoterapie metabolismus MeSH
- nádory farmakoterapie metabolismus MeSH
- nanogely chemie MeSH
- nosiče léků chemie MeSH
- oxidace-redukce MeSH
- pohyb buněk účinky léků MeSH
- systémy cílené aplikace léků metody MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
The aim of this study was to fabricate novel microparticles (MPs) for efficient and long-term delivery of amikacin (AMI). The emulsification method proposed for encapsulating AMI employed low-molecular-weight poly(lactic acid) (PLA) and poly(lactic acid-co-polyethylene glycol) (PLA-PEG), both supplemented with poly(vinyl alcohol) (PVA). The diameters of the particles obtained were determined as less than 30 μm. Based on an in-vitro release study, it was proven that the MPs (both PLA/PVA- and PLA-PEG/PVA-based) demonstrated long-term AMI release (2 months), the kinetics of which adhered to the Korsmeyer-Peppas model. The loading efficiencies of AMI in the study were determined at the followings levels: 36.5 ± 1.5 μg/mg for the PLA-based MPs and 106 ± 32 μg/mg for the PLA-PEG-based MPs. These values were relatively high and draw parallels with studies published on the encapsulation of aminoglycosides. The MPs provided antimicrobial action against the Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae bacterial strains. The materials were also comprehensively characterized by the following methods: differential scanning calorimetry; gel permeation chromatography; scanning electron microscopy; Fourier transform infrared spectroscopy-attenuated total reflectance; energy-dispersive X-ray fluorescence; and Brunauer-Emmett-Teller surface area analysis. The findings of this study contribute toward discerning new means for conducting targeted therapy with polar, broad spectrum antibiotics.
- MeSH
- amikacin aplikace a dávkování chemie MeSH
- antibakteriální látky aplikace a dávkování chemie MeSH
- Escherichia coli účinky léků MeSH
- Klebsiella pneumoniae účinky léků MeSH
- laktáty chemie MeSH
- mikrobiální testy citlivosti MeSH
- molekulová hmotnost MeSH
- nosiče léků chemie MeSH
- polyestery chemie MeSH
- polyethylenglykoly chemie MeSH
- polyvinylalkohol chemie MeSH
- příprava léků metody MeSH
- Pseudomonas aeruginosa účinky léků MeSH
- rozpustnost MeSH
- Staphylococcus aureus účinky léků MeSH
- tobolky MeSH
- uvolňování léčiv MeSH
- velikost částic MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Fused deposition modelling (FDM) is a process of additive manufacturing allowing creating of highly precise complex three-dimensional objects for a large range of applications. The principle of FDM is an extrusion of the molten filament and gradual deposition of layers and their solidification. Potential applications in pharmaceutical and medical fields require the development of biodegradable and biocompatible thermoplastics for the processing of filaments. In this work, the potential of production of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) (P(3HB-co-4HB)) filaments for FDM was investigated in respect to its thermal stability. Copolymer P(3HB-co-4HB) was biosynthesised by Cupriavidus malaysiensis. Rheological and mechanical properties of the copolymer were modified by the addition of plasticizers or blending with poly(lactic acid). Thermal stability of mixtures was studied employing thermogravimetric analysis and rheological analyses by monitoring the time-dependent changes in the complex viscosity of melt samples. The plasticization of P(3HB-co-4HB) slightly hindered its thermal degradation but the best stabilization effect was found in case of the copolymer blended with poly(lactic acid). Overall, rheological, thermal and mechanical properties demonstrated that the plasticized P(3HB-co-4HB) is a potential candidate of biodegradable polymer for FDM processes.
