Different Concentrations Of Riboflavin Research Articles

Improvement of myofibrillar protein gel strength of Scomberomorus niphonius by riboflavin under UVA irradiation.

In this study, effects of different concentrations of riboflavin (0, 0.02, and 0.1 μmol/g protein) on myofibrillar protein (MP, Scomberomorus niphonius) gel were characterized. The gel structure and properties were studied with or without Ultraviolet A (UVA) irradiation. Electron spin resonance results showed that riboflavin produced ·OH under UVA irradiation, which subsequently oxidized the MP. Compared with the control group, the addition of riboflavin with UVA irradiation increased the strength of the MP gel. The rheological results showed that under UVA irradiation, addition of riboflavin facilitated the sol-gel transition between 45 and 52°C, indicating that oxidation led to significant structural changes which in turn resulted in a more compact and uniform gel network. The presence of riboflavin led to increased carbonyl content and decreased sulfhydryl and free amino groups, which decreased the protein solubility and promoted alpha-helical conformational loss in the secondary structure of the MP. These results all indicated that the MP has been oxidized. Electrophoresis revealed that myosin heavy chains were aggregated in the UVA-treated riboflavin-added MP gel, indicating that protein cross-linking has been induced. All the results indicated that the ·OH produced by riboflavin under UVA irradiation oxidized the MP, and improved protein crosslinking and gel properties.

Novel riboflavin/VE-TPGS modified universal dentine adhesive with superior dentine bond strength and self-crosslinking potential

ObjectiveTo modify a universal dentine adhesive with different concentrations of riboflavin and D-Alpha 1000 Succinate polyethylene (VE-TPGS) as a chemical enhancer and to assess the micro-tensile bond strength (24h/12 months), determine resin penetration, measurement of intermolecular interactions and cytotoxicity. Materials and methodsAn experimental adhesive system based on bis-GMA, HEMA and hydrophobic monomer was doped with RF0.125 (RF – Riboflavin) or RF/VE-TPGS (0.25/0.50) and submitted to μTBS evaluation. Resin dentine slabs were prepared and examined using SEM and TEM. Adhesion force was analysed on ends of AFM cantilevers deflection. Quenched peptide assays were performed using fluorescence scanner and wavelengths set to 320nm and 405nm. Cytotoxicity was assessed using human peripheral blood mononuclear cell line. Molecular docking studies were carried out using Schrödinger small-molecule drug discovery suite 2018-2. Data from viable cell results was analyzed using one-way ANOVA. Bond strength values were analysed by two-way ANOVA. Nonparametric results were analyzed using a Kruskal–Wallis test at a 0.05 significance level. ResultsRF/VE-TPGS0.25 groups showed highest bond strength results after 24-h storage in artificial saliva (p<0.05). RF/VE-TPGS0.50 groups showed increased bond strength after 12-months of ageing. RF/VE-TPGS modified adhesives showed appreciable presence of a hybrid layer. Packing fraction indicated solid angle profiles describing well sized density and topology relations for the RF/VE-TPGS adhesives, in particular with the RF/VE-TPGS0.50 specimens. Qualitative analysis of the phenotype of macrophages was prominently CD163+ in the RF/VE-TPGS0.50. Both the compounds showed favourable negative binding energies as expressed in terms of ‘XP GScore’. ConclusionNew formulations based on the incorporation of RF/VE-TPGS in universal adhesives may be of significant potential in facilitating penetration, distribution and uptake of riboflavin within the dentine surface.

Potential for combined delivery of riboflavin and all-trans retinoic acid, from silk fibroin for corneal bioengineering.

Millions of people worldwide suffer from vision impairing conditions resulting from corneal injury or disease. Silk fibroin (SF) is an emerging biopolymer that has been used for several applications including the fabrication of bioengineered corneas and ocular prostheses. To improve the cell response to SF, riboflavin (RF) and all-trans retinoic acid (RA) were coupled onto SF matrices. RF is a photo-initiator that has previously been combined with ultraviolet light to crosslink corneal collagen while RA has been used to regulate the phenotype of corneal stromal cells and their extracellular matrix deposition. Different concentrations of RF and RA were respectively photo-crosslinked and covalently bound through carbodiimide coupling onto 2% SF matrices. The effect of incorporating these molecules on the physical, chemical and mechanical properties of the matrices was evaluated. The biological response of human corneal stromal cells to the matrices was examined using cellular adhesion assays, proliferation assays, cytoskeleton staining, gene expression analysis and immunocytochemical staining. RF and RA both led to changes in the surface nanostructure and hydrophilicity while just RF increased the material stiffness. Cells cultured on the matrices containing both biomolecules displayed improved cellular proliferation, increased GAG deposition and increased expression of keratocyte genes that are normally associated with healthy corneal stromal tissue. These in vitro studies serve as a starting point for the optimization of loading bioactive molecules on SF based matrices for formulating clinically relevant ocular implants.

Identification and clinical significance of Bordetella holmesii isolated from a rare case of bacteremia in Nantong China

Objective To analyze the identification, drug resistance and clinical significance of a rare bacterium of Bordetella holmesii (B.holmesii) to improve its detection and clinical diagnosis and treatment. Methods A strain isolated from a bacteremia case was identified by bacterial culture, biochemical tests and 16S rRNA gene sequencing. Mega 7.0 software was used to conduct a similarity analysis of 16S rRNA gene sequences between the type strains of Bordetella spp. and the isolate, and then a phylogenetic tree was constructed. Antibiotic resistance of the isolate was determined by E-test. Changes in bacterial growth were measured after adding different concentrations of riboflavin or its inhibitor lumiflavin to the culture medium. Results B. holmesii ABD2 was the pathogen causing bacteremia in the immunocompetent patient. It was deposited under the number of CGMCC 1.13721 in China General Microbiological Culture Collection Center (CGMCC), and the 16S rRNA gene sequences were deposited in National Center for Biotechnology Information (NCBI) with the accession number of KT828544.1. Unrooted tree showed that the B. holmesii strain was highly homologous with B. pertussis. Antibiotic susceptibility test showed that the minimum inhibitory concentrations (MIC) of piperacillin, ceftazidime, cefepime, imipenem, meropenem, ciprofloxacin, levofloxacin, gentamicin, amikacin, erythromycin, tetracycline and polymyxin B against the isolate were low, while the MIC values of cefazolin, cefuroxime, cefoxitin, cefotaxime, aztreonam and trimethoprim-sulfamethoxazole were high. Riboflavin accelerated the growth of B. holmesii ABD2, while its inhibitor lumiflavin had an inhibitory effect. Conclusions As B. holmesii is hard to isolate and identify, limited clinical, microbiological and epidemiological data are available. It is an under-recognized pathogen with a considerable amount of information that remains to be studied. Key words: Bordetella holmesii; Bacteremia; Immunocompetence; 16S rRNA gene sequencing; Riboflavin; lumiflavin

Light-induced reactions of methionine and riboflavin in model wine: Effects of hydrolysable tannins and sulfur dioxide

The riboflavin-mediated photo-degradation of methionine in white wine has been related to onset of undesired light-struck taste. This research investigated the effects of different concentrations of riboflavin and methionine, hydrolysable tannins from various sources (nut galls, chestnut and oak woods) and sulfur dioxide on methionine degradation in a model wine exposed to light. Increased methionine concentration resulted in its increased degradation with the consequent formation of volatile sulfur compounds, namely methanethiol, dimethyl disulphide and dimethyl trisulphide. Tannins, especially nut gall tannin, were effective in limiting both methionine degradation and the production of volatile sulfur compounds. Sulfur dioxide enhanced the methionine degradation although the light-struck taste was not perceived when sulfur dioxide concentration was higher than 50 mg/L. In conclusion, the use of hydrolysable tannins can represent a promising tool for protecting white wine against the light-struck taste also limiting the use of sulfur dioxide.

FRET Between Riboflavin and 9-Anthraldehyde Based Fluorescent Organic Nanoparticles Possessing Antibacterial Activity.

The aqueous suspension of fluorescent nanoparticles were prepared by using 9-anthradehdye derivative (AH). The nanoparticles (AHNPs) were characterized using DLS-zeta sizer and SEM techniques. The photo physical properties of nanoparticles and precursor were measured and compared using UV-absorption spectroscopy, fluorescence spectroscopy and fluorescence lifetime studies. The significant overlap between fluorescence spectrum of AHNPs and excitation spectrum of Riboflavin (RF) led us to explore Fluorescence Resonance Energy Transfer (FRET) studies between AHNPs and RF in aqueous medium. The mechanism of FRET from AHNPs to RF discussed on spectral observations, thermodynamic parameters and changes produces in fluorescence lifetime in absence and presence of different concentrations of RF to AHNPs. The limit of detection for RF (0.071µM) is considerably low compared with reported methods. Thus, we explore AHNPs as novel nano probe for quantitative determination of RF in pharmaceutical samples based on FRET study. In addition with this, AHNPs has excellent antibacterial activity than the bulk material for two different bacteria culture viz. E. coli and Bacillus sps. Graphical Abstract 9-anthradehdye based fluorescent nanoparticles (AHNPs) explores as nano probe to detect Riboflavin (RF) in aqueous medium based on Fluorescence Resonance Energy Transfer (FRET) studies. The proposed analytical method successfully applied for quantitative determination of RF in pharmaceutical samples. In addition, with this, AHNPs has excellent antibacterial activity than the bulk material for two different bacteria culture suspension viz. E. coli and Bacillus sps.

Riboflavin deficiency induces a significant change in proteomic profiles in HepG2 cells

Riboflavin deficiency is widespread in many regions over the world, especially in underdeveloped countries. In this study, we investigated the effects of riboflavin deficiency on protein expression profiles in HepG2 cells in order to provide molecular information for the abnormalities induced by riboflavin deficiency. HepG2 cells were cultured in media containing different concentrations of riboflavin. Changes of cell viability and apoptosis were assessed. A comparative proteomic analysis was performed using a label-free shotgun method with LC–MS/MS to investigate the global changes of proteomic profiles in response to riboflavin deficiency. Immunoblotting test was used to validate the results of proteomic approach. The cell viability and apoptosis tests showed that riboflavin was vital in maintaining the cytoactivity of HepG2 cells. The label-free proteomic analysis revealed that a total of 37 proteins showing differential expression (±2 fold, p < 0.05) were identified after riboflavin deficiency. Bioinformatics analysis indicated that the riboflavin deficiency caused an up-regulation of Parkinson’s disease pathway, steroid catabolism, endoplasmic reticulum stress and apoptotic process, while the fatty acid metabolism, tricarboxylic citrate cycle, oxidative phosphorylation and iron metabolism were down-regulated. These findings provide a molecular basis for the elucidation of the effects caused by riboflavin deficiency.

Impact of crosslinking/riboflavin-UVA-photodynamic inactivation on viability, apoptosis and activation of human keratocytes in vitro.

Riboflavin-UVA photodynamic inactivation is a potential treatment alternative in therapy resistant infectious keratitis. The purpose of our study was to determine the impact of riboflavin-UVA photodynamic inactivation on viability, apoptosis and activation of human keratocytes in vitro. Primary human keratocytes were isolated from human corneal buttons and cultured in DMEM/Ham's F12 medium supplemented with 10% fetal calf serum. Keratocytes underwent UVA light illumination (375 nm) for 4.10 minutes (2 J/cm2) during exposure to different concentrations of riboflavin. Twenty-four hours after treatment, cell viability was evaluated photometrically, whereas apoptosis, CD34 and alpha-smooth muscle actin (α-SMA) expression were assessed using flow cytometry. We did not detect significant changes in cell viability, apoptosis, CD34 and α-SMA expression in groups only treated with riboflavin or UVA light. In the group treated with riboflavin-UVA-photodynamic inactivation, viability of keratocytes decreased significantly at 0.1% riboflavin (P<0.01) while the percentage of CD34 (P<0.01 for both 0.05% and 0.1% riboflavin) and alpha-SMA positive keratocytes (P<0.01 and P<0.05 for 0.05% and 0.1% riboflavin, respectively) increased significantly compared to the controls. There was no significant change in the percentage of apoptotic keratocytes compared to controls at any of the used riboflavin concentrations (P = 0.09 and P = 0.13). We concluded that riboflavin-UVA-photodynamic-inactivation decreases viability of myofibroblastic transformation and multipotent haematopoietic stem cell transformation; however, it does not have an impact on apoptosis of human keratocytes in vitro.

In vitro analysis of riboflavin-modified, experimental, two-step etch-and-rinse dentin adhesive: Fourier transform infrared spectroscopy and micro-Raman studies.

To modify two-step experimental etch-and-rinse dentin adhesive with different concentrations of riboflavin and to study its effect on the bond strength, degree of conversion, along with resin infiltration within the demineralized dentin substrate, an experimental adhesive-system was modified with different concentrations of riboflavin (m/m, 0, 1%, 3%, 5% and 10%). Dentin surfaces were etched with 37% phosphoric acid, bonded with respective adhesives, restored with restorative composite–resin, and sectioned into resin–dentin slabs and beams to be stored for 24 h or 9 months in artificial saliva. Micro-tensile bond testing was performed with scanning electron microscopy to analyse the failure of debonded beams. The degree of conversion was evaluated with Fourier transform infrared spectroscopy (FTIR) at different time points along with micro-Raman spectroscopy analysis. Data was analyzed with one-way and two-way analysis of variance followed by Tukey's for pair-wise comparison. Modification with 1% and 3% riboflavin increased the micro-tensile bond strength compared to the control at 24 h and 9-month storage with no significant differences in degree of conversion (P<0.05). The most predominant failure mode was the mixed fracture among all specimens except 10% riboflavin-modified adhesive specimens where cohesive failure was predominant. Raman analysis revealed that 1% and 3% riboflavin adhesives specimens showed relatively higher resin infiltration. The incorporation of riboflavin in the experimental two-step etch-and-rinse adhesive at 3% (m/m) improved the immediate bond strengths and bond durability after 9-month storage in artificial saliva without adversely affecting the degree of conversion of the adhesive monomers and resin infiltration.

Kinetic of Azo Dyes Decolourization by Enterobacteriaceae Species in the Intact Cell Assay System

The method Intact cell was adopted to assess the influencing factors on the process of decolourization ability of the enterobacteriaceaea specie isolates under predisposed environment. Taken into account several ingredient are added individually as well in combinations in the assay reaction. Values of dye decolourized in µg ml -1 with Multi vitanmins solution (MVS) with glucose 42.53; MVS 36.00; Riboflavin (RF) 39.00; Yeast extract 36.00 and B- complex 23.00 from the initial dye of 46.00µg ml -1 were observed , where as glucose, Ascorbic acid, Cysteine, Cetyl-trimethyl-ammonium bromide (CTAB), Sodium molybdate, Biotin, KNO3, NaNO2, folic acid and 1-amino-2- naphtho-4-sulfonic acid (ANSA) does not shows influence on to the process, but, some of them showed inhibitory effect toward the decolourization. It was observed that the riboflavin addition at 19.95 n moles ml -1 in the reaction mixture, rate of decolourization was suddenly change from 0.019 ∆A/minutes to 0.20 ∆A/minutes, which is extremely high by 10 fold fast and subsequently remains faster i.e 0.2 ∆A/minutes without further additional RF in the same assay mixture. Rate of decolourization with different concentrations of riboflavin i.e. 13.0 n moles ml -1 to 59.9 n moles ml -1 showed second order kinetic. This indicates that the minimum amount of RF is essential to trigger the process of decolourization by the intact cells under the assay condition. While five different azo dyes were subjected, showed diverse behavior on to the rate of decolourization. Results of entire study incite on role of riboflavin could to a certain extent act as a redox mediator in the reaction(s) process and electron mediator between intracellular pool to the dye available at periplasmic redox sink @ JASEM

Generation of 8‐hydroxydeoxyguanosine from DNA using rat liver homogenates

In relation to carcinogenesis, aging and other pathologic conditions, urinary 8-hydroxydeoxyguanosine (8OHdG) is widely used as a marker for evaluating the effect of oxidative stress on DNA. Because no reports have described how 8OHdG is generated from DNA in vivo or by biological materials, and how it is excreted into urine, the authors investigated the generation of 8OHdG from DNA, using rat liver homogenate. Oxidatively damaged DNA samples containing different levels of 8OHdG were prepared using ultraviolet irradiation with three different concentrations of riboflavin. Following incubation of damaged DNA samples with rat liver homogenates, the generation of 8OHdG from the DNA was determined using high-performance liquid chromatography with electrochemical detection after ultrafiltration of the incubation mixtures. The generation of 8OHdG was also tested with an anti-8OHdG antibody. The quantity of 8OHdG generated from the DNA by rat liver homogenates was dependent on the 8OHdG levels in the DNA: almost all 8OHdG in the DNA was released as 8OHdG by rat liver homogenates. Generation of 8OHdG correlated with the degradation of DNA. Interestingly, the generated 8OHdG was stable in the presence of rat liver homogenates, whereas deoxyguanosine (dG) rapidly disappeared in the same conditions. Less than 1/10,000 of dG was converted to 8OHdG when dG was incubated with rat liver homogenate. Incubation of 8-hydroxyguanine with rat liver homogenates did not generate 8OHdG. These findings suggest that most of the 8OHdG in DNA is released as 8OHdG during DNA degradation and that, because of its stability, 8OHdG is excreted into urine, thus providing a convenient measure of oxidative damage to DNA.

Influence de la riboflavine sur les besoins globaux en vitamines hydrosolubles chez le puceron du pois, Acyrthosiphon pisum

The effects of different concentrations of riboflavin and that of total water-soluble vitamins in the nutrition of biotypes Saint-Jean and Ile-aux-Coudres of the pea aphid, Acyrthosiphon pisum (Harris), were determined with chemically defined diets. Omission of riboflavin in the diet resulted in a significant increase in growth, reproduction, survival and ingestion rates of the two biotypes. Aphid performance was progressively and significantly improved when the total concentration of 10 vitamins, in the absence of riboflavin, was increased from 0 to 115.6 and 231.1 mg/100 mL of diet; the aphids performed poorly when such increases were accompanied by an increase in riboflavin. Finally, a doubling in the total concentration of the 10 other dietary vitamins (i.e. to 462.2 mg/100 mL), in the absence of riboflavin, resulted in a slightly lowered performance by aphids, whereas, in the presence of riboflavin, aphid performance was significantly reduced. These results indicate that a substantial decrease in dietary riboflavin, or even its omission, and keeping the overall content of the 10 other vitamins to 231.1 mg/100 mL significantly improved the rearing of the two biotypes on such diets over at least two or three successive generations.