Riboflavin Concentration Research Articles

Effects of riboflavin supplementation level on health, performance, and fertility of organic broiler parent stock and their chicks

Data from breeder hens and chicks are particularly critical for the development of vitamin B2 recommendations for organic poultry. To test safe thresholds of supplementation, 100 breeder hens (Hubbard JA 57 K) and ten cockerels were allocated to ten housing groups, with each receiving supplementation of either 4.0 (treatment CON) or 2.5 mg (treatment RED) riboflavin per kg feed. After 15 weeks of experimental feeding, 256 eggs were incubated. From the hatched chicks (Hubbard S757), 192 were allocated to four treatments based on their parents’ treatment (CON- or RED-) and on their own supplementation of either 4.0 (-CON) or 2.5 mg (-RED) riboflavin per kg feed. Two groups of each combination (CON-CON, CON-RED, RED-CON, RED-RED), each with 24 chicks of both sexes, were fattened for nine weeks and slaughtered. Analysis of total riboflavin (sum of native concentrations and supplements) in the parent’s feeds revealed an average, over the 15 weeks, of 7.8 and 5.8 mg per kg feed for CON and RED, respectively. Body weight, plumage integrity, gait appearance, footpad, claw and keel bone integrity, lesion scores, laying performance, and egg mass were found to be of similarly high levels for all hens without any treatment effects. Performance traits of the hens in both treatments reached the specifications of the producer for this genotype. Yolk and albumen riboflavin concentrations were not affected although yolk colour in the RED treatment group became darker. Fertility was not affected, and hatchability was 100 per cent. Total riboflavin concentrations in the broiler diets were on average, over 9 weeks, 8.2 and 6.1 mg/kg for CON and RED, respectively. In chicks, RED treatment of their parents led to significant depressions of feed intake and growth. The RED treatment of the chicks themselves also impaired growth. Growth rates of the CON-CON treatment were in line with the specifications of the producer for this genotype. Plumage scores, footpad integrity and walking appearance of the broilers, and dressing percentage of the carcasses were not affected by treatment. The RED treatment of chicks caused lower spleen and heart weights, while pancreas and liver weights, and liver riboflavin concentrations were not affected. In conclusion, supplementation of 4.0 mg/kg to organic diets did not evoke any health or performance problems for breeder hens or broiler chicks, while a supplementation level of 2.5 mg per kg feed led to reduced weight gains, although without any other riboflavin deficiency symptoms.

Multi-Analyte Sensor Based on Pencil Graphite Electrode for Riboflavin and Pyridoxine Determination

Riboflavin (VB2) and pyridoxine (VB6) are two of the eight water-soluble B vitamins that play an essential role and are correlated in normal function of the body. In the present study, cyclic voltammetry demonstrated that in Britton-Robinson buffer (pH 5.00) VB2 undergoes a quasi-reversible electron transfer reaction and the oxidation of VB6 is irreversible, both electrochemical systems being diffusion controlled at the pencil graphite electrode (PGE) surface. Also, a sensitive and selective voltammetric procedure has been developed for the simultaneous determination of the two compounds using PGE and square wave voltammetry (SWV). Two well-defined oxidation peaks with a voltammetric separation of more than 1.10 V were obtained in Britton-Robinson buffer (pH 5.00). The SWV curves exhibited linear responses with riboflavin and pyridoxine concentrations. Thus, for VB2 two linear concentration ranges of 1.00 · 10−7–5.00 · 10−5 M and 5.00 · 10−5–7.50 · 10−4 M were obtained and in the case of VB6 the linear response was between 2.50 · 10−5 M and 2.50 · 10−3 M. The detection limits for VB2 and VB6 were 7.38 · 10−8 M and 1.10 · 10−5 M, respectively. The applicability of PGE was successfully proved for simultaneous determination of VB2 and VB6 in pharmaceutical tablets with good accuracy and precision.

The Effect of Irradiated Riboflavin in Human Tenon’s Fibroblast – A Study on Cellular Viability

ABSTRACT Purpose/ Aim The main purpose of this work is to study the cellular viability effect of irradiated riboflavin in cultured human tenon fibroblasts. Materials and Methods The tenon tissue was harvested from a patient undergoing strabismus surgery. The human tenon fibroblast cell culture and isolation were performed according to the standard laboratory cell culturing protocol. The cells were divided into three groups: control, treatment with irradiated and non-irradiated riboflavin. There were five different concentrations (0.00156%, 0.003125%, 0.00625%, 0.0125%, 0.025%) in each group of riboflavin. The fibroblasts were treated with riboflavin and the cellular viability was assessed at 24-hour and 48-hour post treatment with MTT 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyl tetrazolium bromide colorimetric assay. The absorbance values were analysed using Magellan microplate reader data analysis. A triplicate of readings was taken. The data were presented as mean ± standard deviation of the triplicates. Statistical analysis was performed with Statistical Package for Social Sciences (SPSS) analysis version 23. Results Irradiated riboflavin caused a concentration-dependent cell death in human tenon fibroblast cell culture (p < .05). The antiproliferative difference between irradiated and non-irradiated riboflavin was significant up to 48 hours (p < .05). Post hoc multiple comparisons showed higher concentrations of irradiated riboflavin (0.0125% and 0.025%) caused more reduction in cellular viability in human tenon fibroblast cells (p < .05). The duration of treatment is not a causative factor in this study. Conclusions This pilot experiment demonstrated that irradiated riboflavin induced cell death in human tenon fibroblast culture in a concentration-dependent manner, but is not time-dependent. Further exploratory investigations should be performed to determine the mechanism of cell death. We postulate that apoptosis occurred in these irradiated riboflavin-treated cells.

Development of nutrient medium for riboflavin biosynthesis by Eremothecium ashbyi ascomycetes

The object of the study is the riboflavin producer Eremothecium ashbyi Guilliermond 1935 VKPM F-340, the subject of the study is the regularities of riboflavin biosynthesis by the E. ashbyi F-340 strain under different cultivation conditions. Riboflavin is an important micronutrient that is a precursor of the coenzymes flavin mononucleotide and flavinadine dinucleotide, it is necessary for biochemical reactions in all living cells. Population growth and an increase in human needs for vitamin-fortified food and agricultural products is the reason for an increase in demand for riboflavin preparations. Considering this, it is important and economically beneficial to improve the technology for the production of vitamin B2. An important factor that affects the yield of the product is the nutrient medium. At present, the influence of agricultural waste on the biosynthesis of riboflavin is being actively studied in the world. However, not all of the studied types of raw materials are typical for the agriculture of this or that country. Therefore, in order to determine whether this direction of research is promising, it is important to check the effect on the biosynthetic activity of the riboflavin producer of the most common wastes of the domestic industry. In this work, this is done on the example of Ukraine. In the course of the study, microbiological (surface and deep cultivation of E. ashbyi F-340), physicochemical (determination of the amount of biomass by the gravimetric method, determination of the concentration of riboflavin by the spectrophotometric method) and mathematical methods were used. The proposed media with the addition of agricultural waste, providing a higher yield of riboflavin compared to conventional media. The influence of different types of agricultural waste on the biosynthesis of riboflavin by the producer E. ashbyi F-340 was evaluated. The efficiency of using sunflower cake as a component of the nutrient medium is shown. The optimal sources of carbon for the nutrient medium with oil cake have been determined, which increases the yield of riboflavin. Due to the large amount of sunflower cake obtained in Ukraine, its use for modifying the nutrient medium in order to increase the yield of riboflavin in the future will lead to a decrease in the cost of the target product due to the use of cheap and ecological raw materials.

Evaluation of the Physiological Corneal Intrastromal Riboflavin Concentration and the Corneal Elastic Modulus After Violet Light Irradiation.

PurposeKeraVio is a corneal crosslinking treatment modality that utilizes violet light (VL)-emitting glasses and topical epithelium-on riboflavin administration. We focus on the new KeraVio protocol without riboflavin. This study aims to quantify the physiological intrastromal concentrations of riboflavin in corneas without riboflavin decreases and evaluate the biomechanics of corneas after VL irradiation.MethodsTwelve human donor corneas were included in this study and randomly categorized into four groups. The corneas underwent four imbibition techniques (physiological riboflavin without drops, epithelial [epi]-on with 0.05% flavin adenine dinucleotide [FAD], epi-off with FAD, and 0.1% riboflavin epi-off). Corneas in the FAD epi-on, FAD epi-off, and riboflavin epi-off groups were instilled with the respective solution every 2 minutes for 30 minutes. An ex vivo experiment was conducted with 24 porcine corneas arranged into three treatment groups and one control group. Corneas in the KeraVio with FAD epi-on group were treated with VL irradiation at 0.31 mW/cm2 for 4.8 hours (5.4 J/cm2) and simultaneously received FAD drops every 30 minutes during the VL irradiation. Corneas in the group with KeraVio without FAD epi-on were only treated with VL irradiation (5.4 J/cm2).ResultsWe identified the original physiological riboflavin of human corneal stroma at a concentration of 0.31 ± 0.03 µg/g, but its value was approximately 39-fold smaller than that in the 0.1% riboflavin epi-off group. The group with KeraVio without FAD and the standard corneal crosslinking group showed a significant increase in biomechanical stability compared with the controls, whereas the elastic modulus in the treated groups was equivalent.ConclusionsWe preliminarily identified physiological riboflavin in human corneas without adding riboflavin drops. The VL exposure may strengthen the corneal biomechanics without requiring the use of additional riboflavin drops.Translational RelevanceWe preliminarily identified physiological riboflavin in the human cornea without adding riboflavin drops. VL irradiation without riboflavin drops may increase the corneal stiffness using physiological riboflavin.

Metarhizium robertsii protease and conidia production, response to heat stress and virulence against Aedes aegypti larvae

Nutritional factors exert significant influence on the growth of entomopathogenic fungi, one of the main agents employed commercially in the biological control of arthropods. Thus, the objective of this work is to optimize the culture medium and solid fermentation time for production of proteases and conidia of Metarhizium robertsii ARSEF 2575 and to evaluate the interference of riboflavin and salts on virulence and resistance to abiotic stress factors. In the first step, nine groups were separated: negative control, positive control, and seven supplementation groups: ammonium nitrate, ammonium chloride, potassium nitrate, sodium nitrate, ammonium sulfate, ammonium phosphate, urea. Sodium nitrate showed significant difference in protease production at the time of 20 days of solid fermentation. Then, different concentrations of sodium nitrate and riboflavin as supplement were evaluated. Response surface methodology demonstrated that riboflavin and sodium nitrate influence proteolytic activity and conidia production, but without synergism. Supplementation of the medium with the optimal concentration of sodium nitrate and riboflavin did not interfere with the germination of conidia without exposure to abiotic stress, but did increase the thermotolerance of conidia. The presence of riboflavin and sodium nitrate at optimal concentrations in the culture medium did not alter fungal virulence with and without exposure to heat stress, varying according to the presence or absence of the supernatant during exposure, evidencing that resistance to heat exposure is multifactorial and dependent on intra- and extracellular factors. Moreover, the supplementation increased the larvicidal activity of the supernatant against Aedes aegypti.

Corneal riboflavin gradients and UV-absorption characteristics after topical application of riboflavin in concentrations ranging from 0.1 to 0.5%

Avoiding damage of the endothelial cells, especially in thin corneas, remains a challenge in corneal collagen crosslinking (CXL). Knowledge of the riboflavin gradients and the UV absorption characteristics after topical application of riboflavin in concentrations ranging from 0.1% to 0.5% could optimize the treatment. In this study, we present a model to calculate the UV-intensity depending on the corneal thickness.Ten groups of de-epithelialized porcine corneas were divided into 2 subgroups. Five groups received an imbibition of 10 min and the other five groups for 30 min. The applied riboflavin concentrations were 0.1%, 0.2%, 0.3%, 0.4% and 0.5% diluted in a 15% dextran solution for each subgroup. After the imbibition process, two-photon fluorescence microscopy was used to determine fluorescence intensity, which was compared to samples after saturation, yielding the absolute riboflavin concentration gradient of the cornea. The extinction coefficient of riboflavin solutions was measured using a spectrophotometer. Combining the obtained riboflavin concentrations and the extinction coefficients, a depth-dependent UV-intensity profile was calculated for each group.With increasing corneal depth, the riboflavin concentration decreased for all imbibition solutions and application times. The diffusion coefficients of 10 min imbibition time were higher than for 30 min. A higher RF concentration and a longer imbibition time resulted in higher UV-absorption and a lower UV-intensity in the depth of the cornea. Calculated UV-transmission was 6 percentage points lower compared to the measured transmission.By increasing the riboflavin concentration of the imbibition solution, a substantially higher UV-absorption inside the cornea is achieved. This offers a simple treatment option to control the depth of crosslinking e.g. in thin corneas, resulting in a lower risk of endothelial damage.

The effect of parasitic ions on riboflavin permeability and cross-linking effectiveness in iontophoresis-assisted scleral cross-linking

The effect of parasitic ions on the results of ultraviolet A (UVA) cross-linking in iontophoresis was still not clear. In this work, the porcine sclera was cross-linked by riboflavin lactate Ringer's solution (group A) and riboflavin normal saline (group B) in vitro, respectively. The concentration of parasitic ions in the solution was calculated. In addition, the average fluorescence intensity, penetration depth and concentration after the introduction of riboflavin and the mechanical properties of cross-linked sclera tissue were measured. The ranges of diffusion coefficient of the two solutions were also calculated, respectively. The results showed that more kinds of parasitic ions were detected in group A compared with group B, while the average fluorescence intensity, penetration depth and concentration of riboflavin and scleral elastic modulus in group B were significantly higher than those in group A when the penetration time was 10 minutes. Besides, the diffusion coefficient of riboflavin in group B was about 1.5 times larger than that in group A. The results suggested that the species of parasitic ions has a great impact on the permeability of riboflavin, and affects the mechanical properties of cross-linked sclera. The above results could provide a reference for improving the efficiency of riboflavin introduction and optimizing the formula of riboflavin in iontophoresis scleral cross-linking.

Vitamin B status and association with antiseizure medication in pregnant women with epilepsy.

Antiseizure medication (ASM) use interacts with vitamin B status in nonpregnant epilepsy populations. We aimed to examine the association between ASM and vitamin B status in pregnant women with epilepsy. We performed a cross-sectional study of pregnancies in women with epilepsy enrolled in the Norwegian Mother, Father and Child Cohort Study from 1999 to 2008. Data on ASM and vitamin supplement use were collected from questionnaires. We analyzed maternal plasma concentrations of ASM and metabolites of folate, including unmetabolized folic acid (UMFA), riboflavin (vitamin B2), pyridoxine (vitamin B6), and niacin (vitamin B3) during gestational weeks 17-19. We included 227singleton pregnancies exposed to ASM with available plasma samples (median maternal age 29years, range 18 to 41years). From the preconception period to gestational week 20, any supplement of folic acid was reported in 208 of pregnancies (94%), riboflavin in 72 (33%), pyridoxine in 77 (35%), and niacin in 45 (20%). High ASM concentrations correlated with high concentrations of UMFA and inactive folate metabolites, and with low concentrations of riboflavin and metabolically active pyridoxine. There was no association between ASM and niacin status. ASM concentrations during pregnancy were associated with vitamin B status in pregnant women with epilepsy. Additional studies are needed to determine the clinical impact of these findings, and to define the optimal vitamin doses that should be recommended to improve pregnancy outcomes.

The performance of a cyanobacterial biomass-based microbial fuel cell (MFC) inoculated with Shewanella oneidensis MR-1

Cyanobacterial biomass is a rich renewable resource for direct bioelectricity generation in biomass-based microbial fuel cells (MFCs). In this study, the performance of a double chamber MFC with Anabaena vaginicola cyanobacterial biomass as the anodic substrate was investigated. The anode chamber was inoculated with a pure culture of Shewanella oneidensis MR-1, and different MFC experiments using pretreated and untreated cyanobacterial biomass substrate with 5 or 20 mg/L riboflavin were conducted. Pretreatment of the biomass and addition of 20 mg/L riboflavin led to the highest current density (366 mA/m3), maximum power density (144 mW/m3), chemical oxygen demand removal efficiency (65%), and coulombic efficiency (5.7%). Increasing the concentration of riboflavin improved bioelectricity generation (1.4–1.7-fold increase in maximum power density), confirming its effectiveness as a redox mediator, and its role in the extracellular electron transfer process of the S. oneidensis MR-1 biofilm. Moreover, pretreating the biomass also enhanced the conversion efficiency of biomass to bioelectricity. This indicated that pretreatment of the biomass influenced the utilization of biomass by S. oneidensis MR-1 cells. These findings can help increasing the potential of using renewable substrates for more efficient bioenergy generation in pure culture MFCs.

Critical Analysis of Corneal Cross-linking (Part-II): Resolving the Controversial Issues (Theory versus Measurements)

Aims: To resolve the controversial issues of UV-light-initiated corneal collagen cross-linking (CXL) by theoretical formulas and measured clinical outcomes.&#x0D; Study Design: Analysis and measured data of CXL.&#x0D; Place and Duration of Study: New Vision Inc, Taipei, between June 2021 and August 2021.&#x0D; Methodology: The controversial issues are addressed and resolved by analytical formulas including: the validation of Bunsen Roscoe law (BRL), the cutoff light intensity, the minimum corneal thickness, the demarcation line depth, the role of oxygen and riboflavin concentration. The overall CXL efficacy is governed by UV-A light intensity, dose, exposure time, mode of exposure (pulsed or CW), the riboflavin concentration, diffusion and drops pre-operation and interoperation administration, the concentration of oxygen in the stromal tissue (pre-op and inter-op), and environmental conditions. The length of the riboflavin presoaking time and viscosity of the riboflavin film also affect the crosslink depth. Analytic formulas are derived for the scaling laws for type-I and type-II efficacy, given by the square root of light intensity, and light dose, respectively.&#x0D; Conclusion: The controversial issues of CXL may be partially resolved via analytic formulas, and compared with measurements. The scaling laws of type-I and type-II efficacy are different and given by analytic formulas. Our formulas also predict the maximum light intensity and the minimum corneal thickness, which are consistent with measurements.

From Farm to Fork: Crickets as Alternative Source of Protein, Minerals, and Vitamins.

Globally, there is growing interest to integrate cricket-based ingredients (flour) into food products to combat food and nutrition insecurity. However, there is lack of information on in-depth nutrient profile of the two cricket species (Scapsipedus icipe and Gryllus bimaculatus), which are the most widely consumed in Africa. Here we determined the nutrient composition of two cricket species and compared them with published records of key animal and plant sources. Our results revealed that the crude protein contents of S. icipe and G. bimaculatus were similar (56.8 and 56.9%, respectively) and comparable to those of animal protein sources. Both cricket species had balanced amino acid profiles that are superior to that of animal and plant sources, except for histidine and cysteine. The protein digestibility of S. icipe and G. bimaculatus ranged between 80 and 88%, which is comparable to that of common plant foods but slightly lower than that of animal proteins. The iron, Zinc, and potassium contents were considerably higher in both cricket species compared to that of plant and animal sources. The calcium contents of both crickets (S. icipe and G. bimaculatus) was superior to that of plant and animal origin except for kidney beans and eggs, respectively. Riboflavin, thiamine, and folic acid concentrations of S. icipe and G. bimaculatus were superior to that of the conventional sources. Vitamin A levels were significantly higher in S. icipe compared to G. bimaculatus. This implies that S. icipe and G. bimaculatus can adequately contribute to our daily required nutrient intake. Thus, integrating cricket flours into ready-to-eat food products would address some of the most pressing nutritional deficiency challenges that many developing countries have to grapple with, particularly high risk to serious health problems such as anemia, poor pregnancy outcomes, hypertension, increased risk of morbidity and mortality, stunted growth and impaired physical and cognitive development. We conclude that edible crickets present unique opportunities for improving food and nutritional insecurity status of both resource-poor and Western populations.

Riboflavin for laying hens fed organic winter diets: effects of different supplementation rates on health, performance and egg quality

ABSTRACT For reasons of aspired naturalness and cost reduction, vitamin additions should be kept low in organic livestock systems. To define safe lower threshold levels of riboflavin supplementation in organic poultry feeding, 135 hybrid layers were allocated to three dietary treatments supplemented with either 4.5 (R4.5), 3.0 (R3.0) or 1.5 mg (R1.5) GMO-free riboflavin added per kg feed. This resulted in total measured concentration averages of 5.0, 4.5, and 3.1 mg kg−1 feed for R4.5, R3.0 and R1.5, respectively. For 18 weeks, each treatment was replicated in three groups of 15 hens. Feed consumption, laying performance, egg quality, riboflavin concentration in the yolk, and clinical health scores were assessed. Two hens per group (six per treatment) were slaughtered in weeks 11 and 18, respectively, for measurement of pancreas, spleen, liver and heart weights, and determination of liver riboflavin. No treatment effects on feed consumption, body weight, laying performance, egg weights, eggshell strength and yolk colour or plumage and keel bone integrity were observed. No symptoms of lameness or footpad damage were found. The riboflavin concentration in egg yolks in R1.5 was lower than in other treatments. Liver riboflavin concentration increased between week 11 and 18 in all treatments, but it was also lower in R1.5. Based on these results, the addition of 3 mg riboflavin kg−1 feed appeared to be sufficient with respect to the health and performance of laying hens. Supplementation with only 1.5 mg kg−1 resulted in some metabolic signs of deficiency. These results apply only to organic winter diets.

The immediate effect of riboflavin and lumichrome on the mitigation of saline stress in the microalga Chlorella sorokiniana by the plant-growth-promoting bacterium Azospirillum brasilense

This study assessed the early effects of riboflavin and lumichrome produced by the bacterium Azospirillum brasilense on mitigating the stress of salinity on the growth and metabolic status of Chlorella sorokiniana. C. sorokiniana was grown in a synthetic growth medium (SGM) at three salt levels (0%, 1.5%, and 3% NaCl) and under four treatments: (1) C. sorokiniana growing alone; (2) C. sorokiniana co-cultured with A. brasilense; (3) C. sorokiniana supplemented with synthetic riboflavin; and (4) C. sorokiniana supplemented with lumichrome, mimicking the riboflavin and lumichrome concentrations naturally produced by A. brasilense. Cell density, total carbohydrates, total and neutral lipids, chlorophyll a and b, and carotenoids contents, proline accumulation, and activity of superoxide dismutase (SOD) were analyzed. Increasing salinity affected the growth rate, cell size, and cell density of C. sorokiniana, induced proline accumulation, and made the total-lipid concentration higher than the carbohydrate concentration. The neutral lipids, chlorophylls, and SOD showed higher values at 1.5% NaCl. Riboflavin and lumichrome showed significant effects on the analyzed variables. Lumichrome at 1.5% salinity induced an early but transient growth response, making carbohydrate production higher than the total lipid production. By contrast, riboflavin enhanced SOD activity and reduced proline accumulation at all the salinity levels. Longer incubation times resulted in effects similar to those of lumichrome in the analyzed metabolites, thus suggesting the conversion of riboflavin into lumichrome. The microalga−bacteria co-cultures at all salinity levels exhibited growth promotion, with consistent effects at the different incubation times and superior magnitude to that presented by lumichrome or riboflavin alone, showing an alleviating effect of the saline stress through osmoregulation with increased carbohydrate concentration and antioxidant activity, but not with increased proline or total lipid concentration. These findings suggest that Azospirillum brasilense can be used as an alternative to mitigate saline stress effects in microalgae cultured in low-salinity waters.

Can UVA-light-activated riboflavin-induced collagen crosslinking be transferred from ophthalmology to spine surgery? A feasibility study on bovine intervertebral disc.

BackgroundCollagen cross-links contribute to the mechanical resilience of the intervertebral disc (IVD). UVA-light-activated riboflavin-induced collagen crosslinking (UVA-CXL) is a well-established and effective ophthalmological intervention that increases the mechanical rigidity of the collagen-rich corneal matrix in Keratoconus. This study explores the feasibility, safety and efficacy of translating this intervention in reinforcing the IVD.MethodsAnnulus fibrosus (AF) cells were isolated from bovine IVDs and treated with different combinations of riboflavin (RF) concentrations (0.05–8 mM) and UVA light intensities (0.3–4 mW/cm2). Metabolic activity (resazurin assay), cell viability (TUNEL assay), and gene expression of apoptosis regulators C-FOS and PT5 were assessed immediately and 24 hours after treatment. Biomechanical effects of UVA-CXL on IVDs were measured by indentation analysis of changes in the instantaneous modulus and by peel-force delamination strength analysis of the AF prior and after treatment.ResultsDifferent intensities of UVA did not impair the metabolic activity of AF cells. However, RF affected metabolic activity (p < 0.001). PT53 expression was similar in all RF conditions tested while C-FOS expression decreased 24 hours after treatment. Twenty-four hours after treatment, no apoptotic cells were observed in any condition tested. Biomechanical characterizations showed a significant increase in the annular peel strength of the UVA-CXL group, when compared to controls of UVA and RF alone (p < 0.05). UVA-CXL treated IVDs showed up to 152% higher (p < 0.001) instantaneous modulus values compared to the untreated control.ConclusionThis is the first study on UVA-CXL treatment of IVD. It induced significantly increased delamination strength and instantaneous modulus indentation values in intact IVD samples in a structure–function relationship. RF concentrations and UVA intensities utilized in ophthalmological clinical protocols were well tolerated by the AF cells. Our findings suggest that UVA-CXL may be a promising tool to reinforce the IVD matrix.

Extracellular riboflavin induces anaerobic biofilm formation in Shewanella oneidensis

BackgroundSome microorganisms can respire with extracellular electron acceptors using an extended electron transport chain to the cell surface. This process can be applied in bioelectrochemical systems in which the organisms produce an electrical current by respiring with an anode as electron acceptor. These organisms apply flavin molecules as cofactors to facilitate one-electron transfer catalyzed by the terminal reductases and in some cases as endogenous electron shuttles.ResultsIn the model organism Shewanella oneidensis, riboflavin production and excretion trigger a specific biofilm formation response that is initiated at a specific threshold concentration, similar to canonical quorum-sensing molecules. Riboflavin-mediated messaging is based on the overexpression of the gene encoding the putrescine decarboxylase speC which leads to posttranscriptional overproduction of proteins involved in biofilm formation. Using a model of growth-dependent riboflavin production under batch and biofilm growth conditions, the number of cells necessary to produce the threshold concentration per time was deduced. Furthermore, our results indicate that specific retention of riboflavin in the biofilm matrix leads to localized concentrations, which by far exceed the necessary threshold value.ConclusionThis study describes a new quorum-sensing mechanism in S. oneidensis. Biofilm formation of S. oneidensis is induced by low concentrations of riboflavin resulting in an upregulation of the ornithine-decarboxylase speC. The results can be applied for the development of strains catalyzing increased current densities in bioelectrochemical systems.

Photochemical reaction of superoxide radicals with 1-naphthol

The photochemical reactions between 1-naphthol (1-NP) and the superoxide anion radical (O2•–) were investigated in detail by using 365 nm UV irradiation. The results showed that the conversion rate of 1-NP decreased with the increase of the initial concentration of 1-NP, whereas by increasing the pH and riboflavin concentration, the photochemical reaction was accelerated. The second-order reaction rate constant was estimated to be (3.64 ± 0.17) × 108 L mol−1s−1. The major photolysis products identified by using gas chromatography – mass spectrometry (GC–MS) were 1,4-naphquinone and 2,3-epoxyresin-2,3-dihydro-1,4-naphquinone, and their reaction pathways were also discussed. An atmospheric model showed that both the bulk water reaction and the heterogeneous surface reaction deserve attention in atmospheric aqueous chemistry.

Unraveling the mechanism of efficient adsorption of riboflavin onto activated biochar derived from algal blooms

Riboflavin is commercially produced primarily by bio-fermentation. Nonetheless, purification and separation are particularly complex and costly. Adsorption from the fermentation liquor is an alternative riboflavin separation technology during which a cost-efficient adsorbent is highly desired. In this study, a low-cost activated algal biomass-derived biochar (AABB) was applied as an adsorbent to efficiently adsorb riboflavin from an aqueous solution. The adsorption capacity of riboflavin on AABB increased with the increase in pyrolysis temperature and initial riboflavin concentration. The adsorption isotherms were well described by the Freundlich and Langmuir models. The AABB displayed excellent adsorption performance and its maximum adsorption capacity was 476.9 mg/g, which was 6.8, 6.8, and 5.2 times higher than that of laboratory-prepared activated rape straw biochar, activated broadbean shell biochar and commercial activated carbon, respectively, which was mainly ascribed to its larger specific surface area and abundant functional groups. The mass transfer model results showed that mass transfer resistance was dependent on both the film mass transfer and porous diffusion. Raman and Fourier transform-infrared spectra confirmed the presence of π-π interactions and hydrogen bonding between riboflavin and the AABB. The adsorption of riboflavin onto AABB was a spontaneous process, which was dominated by van der Waals forces. These results will be beneficial for developing effective riboflavin recovery technologies and simultaneously utilizing waste algal blooms.

The Effect of Sodium Iodide on Stromal Loading, Distribution and Degradation of Riboflavin in a Rabbit Model of Transepithelial Corneal Crosslinking.

PurposeTo evaluate effects of sodium iodide (NaI) on riboflavin concentration in corneal stroma before and during ultraviolet A (UVA) light exposure using a novel transepithelial corneal collagen crosslinking (CXL) procedure (EpiSmart CXL system, CXL Ophthalmics, Encinitas CA).MethodsRiboflavin solutions with NaI (Ribostat, CXL Ophthalmics, Encinitas CA) and without NaI were used for CXL in rabbits using EpiSmart. A pilot study determined sufficient riboflavin loading time. Four rabbits were dosed and monitored. Riboflavin fluorescence intensity was assessed from masked slit-lamp photos. A 12 min loading time was selected. Sixteen additional rabbits received the two formulae in contralateral eyes for CXL. Riboflavin uptake was assessed at 0, 10, 15, 20, 25, and 30 min of UVA exposure using a scale for riboflavin fluorescence previously validated against stromal concentration. Post sacrifice, corneal stromal samples were analyzed for concentrations of riboflavin and riboflavin 5ʹ-phosphate.ResultsEyes dosed with NaI riboflavin had higher riboflavin grades compared to eyes dosed with the NaI-free riboflavin formulation immediately after riboflavin loading and persisting throughout UVA exposure, with significantly higher (P < 0.01 to < 0.05) riboflavin grades from 15 through 25 min of UVA exposure. Riboflavin grades decreased more slowly in eyes dosed with NaI riboflavin through 25 minutes of UVA exposure. Minor conjunctival irritation was noted with or without NaI.ConclusionThe addition of NaI to riboflavin solution is associated with increased riboflavin concentration in corneal stroma throughout a clinically relevant time course of UVA exposure. This effect may be a combination of enhanced epithelial penetration and reduced riboflavin photodegradation and should enhance intrastromal crosslinking.

Collagen Structural Changes in Rat Tarsus After Crosslinking.

PurposeSurgery is the standard treatment for floppy eyelid syndrome, but crosslinking (CXL) tarsus has recently been proposed as an alternative. To the best of our knowledge, this study is the first to use second-harmonic generation (SHG) microscopy to examine tarsal collagen ex vivo before and after photo-activated crosslinking. To quantify crosslinking, this study examined fluorescence recovery after photobleaching (FRAP), which indirectly measures tissue stiffness.MethodsUpper eyelid tarsal plates were dissected from 21 Sprague–Dawley rats (total of 42 tarsal plates). Six normal plates were sent for histopathology and SHG imaging; the remaining 36 were crosslinked with phosphate-buffered saline (PBS) alone or riboflavin in PBS (concentrations of 0.1%, 0.3%, and 0.5%). Tissues were irradiated with 365-nm ultraviolet A light (power, 0.45 mW/cm2) for 30 minutes and immediately underwent SHG microscopy. Stiffness was indirectly measured with FRAP using fluorescein isothiocyanate (FITC)–dextran.ResultsSHG imaging of normal tarsus showed that the organization of collagen bundles is complex and varies greatly depending on location. After crosslinking with high-concentration riboflavin (0.5%), collagen fibers showed clear structural changes, becoming more densely packed and wavier compared to control. FRAP half-time to fluorescence recovery was significantly increased (P < 0.05), indirectly indicating increased tissue stiffness. No structural changes were observed after crosslinking with lower riboflavin concentrations of 0.1% and 0.3%.ConclusionsThis is the first report of SHG microscopy used to image tarsus collagen before and after crosslinking. These results highlight collagen structural changes, with effects on tissue stiffness indirectly confirmed by FRAP.Translational RelevanceCollagen fibers in the tarsus may be a therapeutic target for crosslinking in order to treat symptomatic floppy eyelid syndrome.