Fluorescent Advanced Glycation End-products Research Articles

Revealing the Hypoglycemic Effect of Red Yeast Rice: Perspectives from the Inhibition of α-Glucosidase and the Anti-Glycation Capability by Ankaflavin and Monascin

Red yeast rice dietary supplements have been proven to ameliorate hyperglycemia, but the mechanism was unclear. In this work, ankaflavin (AK) and monascin (MS), as typical pigments derived from red yeast rice, were found to exert noteworthy inhibitory ability against α-glucosidase, with an IC50 of 126.5 ± 2.5 and 302.6 ± 2.5 μM, respectively, compared with acarbose (IC50 = 341.3 ± 13.6 μM). They also exhibited mixed-type inhibition of α-glucosidase in vitro and caused fluorescence quenching through the static-quenching process. Molecular-docking studies indicated that AK and MS bind to amino acid residues outside the catalytic center, which induces structural changes in the enzyme, thus influencing its catalytic activity. The anti-glycation ability of Monascus-fermented products was evaluated, and they exhibited a high inhibition rate of 87.1% in fluorescent advanced glycation end-product formation at a concentration of 0.2 mg mL−1, while aminoguanidine showed a rate of 75.7% at the same concentration. These results will be significant in broadening the application scope of Monascus pigments, especially AK and MS, in treating type 2 diabetes.

Quercetin decreases fructose drinking in model of fructose-induced insulin resistance. Unexpected uric acid and TBARS lowering effect of methyl cellulose vehicle and fructose combination.

The aim of this study was to improve insulin sensitivity in fructose-treated animals by ingestion of flavonoid quercetin. Several signs of insulin resistance have been developed in rats by drinking 10% fructose solution for 9 weeks. The effect of 6-week-gavage-administrated quercetin (20 mg/kg/day in 1% methyl cellulose solution) was monitored. Rats of the control groups received methyl cellulose vehicle as well. The most striking result of the quercetin treatment was the normalization of the fructose solution drinking to the level of drinking water intake. In addition, quercetin supplementation considerably decreased the plasma glucose and Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) index in rats consuming fructose. Surprisingly, fructose ingestion did not elevate plasma uric acid, thiobarbituric acid reactive substances, nitrotyrosine, or advanced glycation end products fluorescence. Instead, a reduction of the above parameters was observed. In summary, these results indicate that quercetin supplementation reduces fructose drinking and decreases plasma glucose and the HOMA-IR index. Furthermore, methyl cellulose, in combination with fructose, causes uric acid - lowering, antioxidant and anti-glycation effects. Thus, methyl cellulose possibly shifts fructose metabolism in favor of the utilization of antioxidant features of fructose. Our results call for using methyl cellulose in sweetened beverages and other sweetened food.

Antiglycating Effects of Spirulina platensis Aqueous Extract on Glucose-Induced Glycation of Bovine Serum Albumin.

Glucose, the predominant carbohydrate in the human body, initiates nonenzymatic reactions in hyperglycemia, potentially leading to adverse biochemical interactions. This study investigates the interaction between glucose and Bovine Serum Albumin (BSA), along with the protective effects of Spirulina platensis PCC 7345 aqueous extract. Phycobiliproteins (phycocyanin, phycoerythrin, and allophycocyanin) in the extract were quantified using spectrophotometry. The extract's anti-glycation potential was assessed by analyzing its effects on albumin glycation, fluorescent advanced glycation end products (AGEs), thiol group oxidation, and β-amyloid structure generation. Additionally, its antidiabetic potential was evaluated by measuring α-amylase and α-glucosidase enzyme inhibition. Results indicate that the Spirulina extract significantly mitigated ketoamine levels, fluorescence, and protein-carbonyl production induced by glucose, demonstrating a 67.81% suppression of AGE formation after 28 days. Moreover, it effectively inhibited amyloid formation in BSA cross-linkages. These findings suggest the potential of S. platensis as an anti-glycation and antidiabetic agent, supporting its consideration for dietary inclusion to manage diabetes and associated complications.

Effects of fibrillation combined with glycation by different chain length sugars on structural and functional properties of soybean protein

The key objective of this study was to investigate the effects of fibrillation combined with glycation of soybean protein isolate (SPI) on the protein structure and functional properties. SPI fibrils alone, glycated proteins and glycated fibrils with different chain lengths (glucose–G, maltose–M and dextran–D) were prepared and characterized by spectroscopy and microscopy methods. The results of the O-phthalaldehyde assay, ultraviolet–visible spectroscopy and fluorescent advanced glycation endproducts indicated the degree of glycation. The thioflavin T fluorescence assay, transmission electron microscopy (TEM) and Fourier-transform infrared (FTIR) spectroscopy results demonstrated that the fibril morphology improved, and more flexible and aggregated fibrils were produced after glycation, further improving the protein functional properties. With EAI and ESI values of 37.31 m2/g and 33.80%, respectively, the emulsifying properties of SPI fibrils glycated by dextran were higher than the others. Foam properties and antioxidant activity were highest for the fibrils glycated by glucose. These results are of scientific significance for in-depth understanding and reasonable regulation of protein functional properties for industrial production in a complex food system.

Mentha piperita silver nanoparticle-loaded hydrocolloid film for enhanced diabetic wound healing in rats.

To investigate the role of Mentha piperita silver nanoparticle-loaded carbopol gel for enhanced wound healing in a diabetic rat model. This research further aims to explore bioactive compounds derived from Mentha piperita obtained from high altitude. Methanolic extracts of Mentha piperita (MP), Mentha spicata (MS) and Mentha longifolia (ML) were used to synthesise silver nanoparticles (AgNP). AgNP synthesis was confirmed by ultraviolet-visible (UV-Vis) spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The antioxidant activity was assessed by 2, 2-diphenyl-1-picrylhydrazyl (DDPH) assay. Antiglycation potential was determined by measuring the fluorescent advanced glycation end products. The bioactive compound identified in the Mentha piperita methanolic (MPM) fraction through electrospray ionisation tandem mass spectrometric analysis (ESI-MS) was responsible for the highest antiglycation. The effects of MPM and MPM.AgNP-loaded Carbopol (Sanare Lab, India) on wound healing were compared in male, alloxan-induced, diabetic albino rats (200-250g), divided into control and treated groups. Effects on wound healing were assessed via histopathology. UV-Vis and FTIR confirmed NP synthesis with peaks for flavonoids and polyphenols. SEM and XRD explored the cubical, 30-63nm crystalline NP. The maximum antioxidant and antiglycation potential was observed in order of; MP.AgNP>MS.AgNP>ML.AgNP. The highest antioxidant activity was observed by methanolic and aqueous MP.AgNPs (88.55% and 83.63%, respectively) at 2mg.ml-1, and (75.16% and 69.73%, respectively) at 1mg.ml-1, compared to ascorbic acid (acting as a positive control, 90.01%). MPM.AgNPs demonstrated the best antiglycation potential of 75.2% and 83.3% at 1mg.ml-1 and 2mg.ml-1, respectively, comparable to positive control (rutin: 88.1%) at 14 days post-incubation. A similar trend was observed for antimicrobial activity against Bacillus subtilis, Micrococcus luteus and Escherichia coli with an inhibition zone of 21mm, 21.6mm and 24.6mm. Rosmarinic acid was the active compound present in Mentha piperita, as identified by ESI-MS. MPM.AgNP-loaded Carbopol resulted in 100% wound closure compared with control at 20 days post-wounding. In the treatment group, re-epithelialisation was achieved by day 18, compared with 25 days for the positive control group. MPM.AgNP-loaded Carbopol demonstrated safer and more effective biological properties, hence accelerating the diabetic excision wound healing process in alloxan-induced diabetic rats.

Inhibition of fluorescent advanced glycation end-products by ferulic acid and chlorogenic acid: A fluorescence spectroscopy and molecular docking study

The effects of ferulic acid (FA) and chlorogenic acid (CGA) on fluorescent advanced glycation end-products (fAGEs) were investigated through the Maillard reaction. Model systems were incubated with different concentrations (0, 10, 20, 40, 80, and 160 μg/mL) of FA or CGA at 55 °C for 12 h, and fluorescence intensity of advanced glycation end-products (AGEs) was measured. After adding 160 μg/mL FA and CGA, the fluorescence intensity of AGEs was suppressed by 15.988% ± 0.556% and 16.544% ± 0.275%, respectively (P < 0.05). Compared with control group, free amino groups levels were the highest in model systems containing 160 μg/mL FA (0.096 ± 0.011 mg l-leucine equivalent/mL) and CGA (0.130 ± 0.000 mg l-leucine equivalent/mL) (P < 0.05). Carbonyls content decreased in a dose-dependent fashion with increasing concentrations of FA and CGA (P < 0.05). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results demonstrated that FA and CGA could impede the generation of bovine serum albumin (BSA)-glucose cross-linking substances. Spectroscopy results indicated that the binding constant of BSA-CGA was higher than that of BSA-FA. Meanwhile, molecular docking results suggested that the binding affinity of CGA to BSA was greater than to FA. These results suggested that CGA had a stronger inhibitory effect on the fluorescence intensity of AGEs. Therefore, FA and CGA may be potent natural antioxidants in mitigating the accumulation of AGEs during food thermal processing.

Formation of Maillard reaction products in a model bread system of different gluten-free flours.

In this study, we determined the levels of Maillard-type products (furosine, free fluorescent advanced glycation end products, 3-deoxyglucosone, methylglyoxal, and N-ε-carboxymethyllysine) in a model gluten-free bread made of wholegrain white, brown, wild, red, and black rice flour, as well as of yellow and purple corn flour. The total protein, lysine, arginine, and cysteine contents were found to relate directly to the formation of Maillard-type products. The malvidin, ferulic, syringic, sinapic, and p-coumaric acids present in gluten-free breads were effective against the formation of furosine, fluorescent compounds, and dicarbonyls, but were ineffective in mitigating CML. Although the bread formulated with purple corn flour had the lowest levels of furosine and fluorescent compounds, this formulation led to an increase in N-ε-carboxymethyllysine and adversely affected the bread's aroma, on account of the presence of polyphenols responsible for a bitter aroma, which were at their highest concentration (1942.34µg/g) in this bread.

Multiphoton imaging of the monosachharide induced formation of fluorescent advanced glycation end products in tissues.

We studied the in vitro rate of fluorescent advanced glycation end products (fAGEs) formation with multiphoton microscopy in different porcine tissues (aorta, cornea, kidney, dermis, and tendon). These tissues were treated with d-glucose, d-galactose, and d-fructose, three primary monosaccharides found in human diets. We found that the use of d-fructose resulted in the highest glycation rate, followed by d-galactose and then d-glucose. Moreover, compared to non-collagen tissue constituents such as elastic fibers and cells, the rate of tissue glycation was consistently higher in collagen, suggesting that collagen is a more sensitive target for fAGE formation. However, we also found that collagen in different tissues exhibits different rates of fAGE formation, with slower rates observed in tightly packed tissues such as cornea and tendon. Our study suggests that for fAGE to be developed into a long-term glycemic biomarker, loosely organized collagen tissues located in the proximity of vasculature may be the best targets.

Anti-glycation and phytochemical properties of cinnamon stem-bark water extract

The study evaluated the ability of cinnamon stem-bark water extract (CWE) to break established cross-links formed between proteins and advanced glycation end-products (AGEs) as well as its anti-glycation effect. The extract showed a dose-dependent anti-glycation effect against total fluorescent AGEs (FAGEs) derived from both glucose and fructose. CWE recorded a uniform IC50 value of 0.13 mg/mL for both glucose and fructose-derived FAGEs while aminoguanidine, a well-known synthetic anti-glycative agent gave IC50 values of 0.14 mg/mL for the glucose and 0.17 mg/mL for the fructose derived FAGEs. The anti-glycative effect of CWE was also significantly higher than aminoguanidine against total immunogenic AGE (TIAGEs) in both sugar models (p &lt; 0.001). While CWE and aminoguanidine showed weak protein cross-link breaking activity selectively on fructose-derived protein cross-links already formed, none whatsoever was detected exerted on the established glucose-derived protein cross-links. Phytochemical screening revealed the presence of several important secondary metabolites which may have contributed to the anti-glycative effect of CWE. Gas chromatography mass spectrometry enabled the identification of (+)-alpha-tocopherol acetate, a chain breaker, in the methanol fraction of CWE. Although CWE showed great potential for inhibition of formation of major types of AGEs, it appears to be poor in breaking established cross-links formed between proteins and AGEs.

Molecularly imprinted thermosensitive probe based on fluorescent advanced glycation end products to detect α-dicarbonyl compounds and inhibit pyrraline formation.

A thermal-sensitive molecularly imprinted optosensing probe based on fluorescent advanced glycation end products (AGEs) was prepared by one-pot hydrothermal synthesis. Carbon dots (CDs) derived from fluorescent AGEs were used as the luminous centers, while molecularly imprinted polymers (MIPs) were wrapped outside of theCDs to form specific target recognition sites to highly selectively adsorbthe intermediate product of AGEs of 3-deoxyglucosone (3-DG). Thermosensitive N-isopropylacrylamide (NIPAM) was combined with acrylamide (AM) as co-functional monomers, and ethylene glycol dimethacrylate (EGDMA) was chosen as across-linker for targeting identification and detection of 3-DG. Under optimal conditions, the fluorescence of MIPs could be gradually quenched with the adsorption of 3-DG on the surface of MIPs in the linear range of 1-160μg/L, and the detection limit was 0.31μg/L. The spiked recoveries of MIPs ranged from 82.97 to 109.94% in two milk samples, and the relative standard deviations were all less than 1.8%. In addition, the inhibition rate for non-fluorescent AGEs of pyrraline (PRL) was 23% by adsorbing 3-DG in the simulated milk system of casein and D-glucose, indicating that temperature-responsive MIPs not only could detect the dicarbonyl compound 3-DG quickly and sensitively, but also had an excellent inhibitory effect on AGEs.

Germ-Free C57BL/6 Mice Have Increased Bone Mass and Altered Matrix Properties but Not Decreased Bone Fracture Resistance.

The gut microbiome impacts bone mass, which implies a disruption to bone homeostasis. However, it is not yet clear how the gut microbiome affects the regulation of bone mass and bone quality. We hypothesized that germ-free (GF) mice have increased bone mass and decreased bone toughness compared with conventionally housed mice. We tested this hypothesis using adult (20- to 21-week-old) C57BL/6J GF and conventionally raised female and male mice (n = 6-10/group). Trabecular microarchitecture and cortical geometry were measured from micro-CT of the femur distal metaphysis and cortical midshaft. Whole-femur strength and estimated material properties were measured using three-point bending and notched fracture toughness. Bone matrix properties were measured for the cortical femur by quantitative back-scattered electron imaging and nanoindentation, and, for the humerus, by Raman spectroscopy and fluorescent advanced glycation end product (fAGE) assay. Shifts in cortical tissue metabolism were measured from the contralateral humerus. GF mice had reduced bone resorption, increased trabecular bone microarchitecture, increased tissue strength and decreased whole-bone strengththat was not explained by differences in bone size, increased tissue mineralization and fAGEs, and altered collagen structure that did not decrease fracture toughness. We observed several sex differences in GF mice, most notably for bone tissue metabolism. Male GF mice had a greater signature of amino acid metabolism, and female GF mice had a greater signature of lipid metabolism, exceeding the metabolic sex differences of the conventional mice. Together, these data demonstrate that the GF state in C57BL/6J mice alters bone mass and matrix properties but does not decrease bone fracture resistance. © 2023 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).

The influence of piperine on oxidation-induced porcine myofibrillar protein gelation behavior and fluorescent advanced glycation end products formation in model systems

This study investigated the effects of piperine on oxidation-induced porcine myofibrillar protein (MP) gelation behavior and fluorescent advanced glycation end products (fAGEs) formation. Model systems were prepared, lipid oxidation, MP gelling behavior, and fAGEs content were determined daily. The results indicated that lipid oxidation, carbonyl content, S0, cooking loss, and tryptophan fluorescence intensity of MP significantly decreased, whereas gel strength, WHC, and whiteness markedly increased as the concentration of piperine increased (from 0 to 30 μM/g protein), indicating that piperine could reduce lipid oxidation and oxidative damage to MP. The fluorescence intensity of fAGEs markedly decreased (P < 0.05), from 93.1 ± 4.4 to 61.2 ± 3.0, as the concentration of piperine increased from 0 μM/g protein to 30 μM/g protein after 5 days of incubation. These results in model systems suggest that the presence of piperine has an important role in the inhibition of MP oxidation and fAGEs formation.

Fluorescent advanced glycation end products in type 2 diabetes and its association with diabetes duration, hemoglobin A1c, and diabetic complications.

Fluorescent advanced glycation end products (fAGEs) are generated through the Maillard reaction between reducing sugars and amino compounds. fAGEs accumulation in human bodies have been confirmed to be related to many chronic diseases. To date, the correlations between serum fAGEs levels and clinical parameters or carotid intima media thickness (CIMT) in patients with T2DM remain unclear. Thus, this study aimed to investigate the relationship between serum AGEs levels and clinical parameters or CIMT in patients with T2DM. A total of 131 patients with diabetes and 30 healthy controls were enrolled. Patients were divided into three groups according to diabetes duration, including ≤5, 5-10, and ≥10 years. Serum fAGEs, protein oxidation products, clinical parameters, and CIMT were determined. The result showed that levels of fAGEs and protein oxidation products increased with the increasing duration of diabetics. Pearson correlation coefficients of fAGEs versus hemoglobin A1c (HbA1c) were >0.5 in patients with diabetes duration ≥10 years. A continued increase in fAGEs might cause the increase of HbA1c, urinary albumin/creatinine ratio (UACR) and CIMT in patients with T2DM. Our study suggested that levels of fAGEs could be considered as an indicator for duration of diabetics and carotid atherosclerosis. Diabetes duration and smoking might have a synergistic effect on the increment of fAGEs levels, as evidence by the results of correlation analysis in patients with long-duration diabetics (≥10 years) and smoking. The determination of fAGEs might be helpful to advance our knowledge on the overall risk of complications in patients with T2DM.

D-Ribose-Induced Glycation and Its Attenuation by the Aqueous Extract of Nigella sativa Seeds.

Background and Objectives: Glycation and oxidative stress are the major contributing factors responsible for diabetes and its secondary complications. Aminoguanidine, a hydrazine derivative, is the only approved drug that reduces glycation with its known side effects. As a result, research into medicinal plants with antioxidant and antiglycation properties is beneficial in treating diabetes and its consequences. This investigation aimed to examine the efficacy of the aqueous extract of Nigella sativa seeds against the D-ribose-induced glycation system. Materials and Methods: The suppression of α-amylase and α-glucosidase enzymes were used to assess the antidiabetic capacity. UV-Visible, fluorescence, and FTIR spectroscopy were used to characterize the Nigella sativa seed extract and its efficacy in preventing glycation. The inhibition of albumin glycation, fluorescent advanced glycation end products (AGEs) formation, thiol oxidation, and amyloid formation were used to evaluate the extracts' antiglycation activity. In addition, the extent of glycoxidative DNA damage was analyzed using agarose gel electrophoresis. Results: The IC50 for the extract in the α-amylase and α-glucosidase enzyme inhibition assays were approximately 1.39 ± 0.016 and 1.01 ± 0.022 mg/mL, respectively. Throughout the investigation, it was found that the aqueous extract of Nigella sativa seeds (NSAE) inhibited the level of ketoamine, exerted a considerable drop in fluorescence intensity, and reduced carbonyl production and thiol modification when added to the D-ribose-induced glycation system. In addition, a reduction in the BSA-cross amyloid formation was seen in the Congo red, thioflavin T assay, and electrophoretic techniques. NSAE also exhibited a strong capability for DNA damage protection. Conclusion: It can be concluded that Nigella sativa could be used as a natural antidiabetic, antiglycation treatment and a cost-effective and environmentally friendly source of powerful bioactive chemicals.

Accumulation of fluorescent advanced glycation end products and carboxymethyl-lysine in human cortical and trabecular bone

Chemical crosslinks known as advanced glycation end-products (AGEs) are associated with increased bone fracture risk and deteriorated bone mechanical properties. However, measurement of bone AGEs via ex vivo and in vitro methods has been limited to quantification of bulk fluorescent AGEs (fAGEs) and pentosidine only, which is a crosslinking fluorescent AGE. However, a non-crosslinking and non-fluorescent AGE such as carboxymethyl-lysine (CML) is found to be 40–100 times higher in quantity than pentosidine, but only one previous study has reported it in cortical bone, and one study reported it in trabecular bone. In our study, we wanted to investigate if accumulation of CML differs in cortical and trabecular compartments and if they are more strongly associated with bone mechanical properties than with fAGEs. We hypothesized that CML and fAGEs level would be higher in the trabecular compartment and show negative correlations to mechanical properties in cortical and trabecular bone. We obtained human cadaveric cortical and trabecular bone specimens, induced the formation of AGEs via the established in vitro ribosylation method, imaged specimens by microcomputed tomography to assess specimen geometry and microarchitecture, and mechanically tested cortical specimens by cyclic reference point indentation and fracture toughness tests and trabecular specimens by compression tests, followed by measurement of fAGEs and CML. fAGEs were 22 % higher in cortical bone (687 ± 44.8 ng Q/mg collagen) compared to trabecular bone (859 ± 317.1 ng Q/mg collagen), whereas CML levels were found to be 148 % higher in trabecular bone (6189.9 ± 866 ng/mg of protein) compared to cortical bone (924.6 ± 576.3 ng/mg of protein). Pooling the specimens from both the control and ribose groups, Spearman correlation analysis indicated that CML levels, but not fAGEs, are moderately associated with cortical porosity (r = +0.505, p ≤ 0.05) and mechanical properties such indentation depth (r = +0.460, p ≤ 0.05), total indentation depth (r = +0.440, p ≤ 0.05), and average energy dissipated (r = +0.465, p ≤ 0.05) in cortical bone. fAGEs showed a trend towards negative association with crack propagation toughness in cortical bone (r = −0.365, p = 0.055). No significant correlations were observed between CML and microarchitecture or mechanical properties in trabecular bone. CML levels were also associated with fAGEs in cortical bone (r = +0.596, p ≤ 0.05) but not in trabecular bone. Our preliminary findings indicate that CML, a non-crosslinking AGE, may affect bone material and mechanical properties differently than bulk fluorescent AGEs, given the higher accumulation of CML in each bone compartment. This study provides direction to future studies to quantify crosslinking and non-crosslinking AGEs separately as their effect on material and mechanical properties may be different and it would help identify better biomarkers for bone strength prediction.

Phytochemical profile, anti-glycation effect, and advanced glycation end-products protein cross-link breaking ability of Sclerocarya birrea stem-bark crude extracts

Introduction: Sclerocarya birrea stem-bark is widely used for the treatment of many medical conditions. Advanced glycation end-products (AGEs) are implicated in the pathogenesis of vascular complications of diabetes mellitus. The study, other than phytochemical composition, evaluated the anti-glycation and AGEs-protein cross-link breaking effects of S. birrea stem-bark extracts. Methods: Different S. birrea extracts and aminoguanidine (used as control) were incubated with bovine serum albumin (BSA) and glucose/fructose at 37oC for 40 days. Amounts of fluorescent AGEs (FAGEs) and immunogenic AGEs formed were determined. Anti-glycation activity percentage of each extract and aminoguanidine was calculated. Their AGEs-protein cross-link breaking abilities were also assessed. Standard techniques were employed for phytochemical screening. Volatile compounds were identified by means of gas chromatography mass spectrometry (GC-MS). Results: S. birrea stem-bark n-hexane extract was statistically more effective than aminoguanidine against the formation of total immunogenic AGEs (P&lt;0.05). For FAGEs, ethyl acetate, methanol, and water extracts exerted significantly higher anti-glycation effects than aminoguanidine (P&lt;0.001). Methanol extract exhibited the highest anti-glycation effect with an average IC50 value of 0.142 mg/mL against FAGEs. All extracts were effective in releasing BSA from the preformed collagen-AGEs-BSA cross-links. GC-MS enabled the identification of many biologically important compounds, including campesterol, stigmasterol, and 1-heptatricontanol. Conclusion: S. birrea stem-bark has a potential for usage in the management of complications in uncontrolled glucose metabolism.

2. The association between lumbar bone mineral density and advanced glycation end products derived from confocal fluorescence microscopy: a prospective investigation of bone biopsies in patients undergoing lumbar spinal fusion surgery

2. The association between lumbar bone mineral density and advanced glycation end products derived from confocal fluorescence microscopy: a prospective investigation of bone biopsies in patients undergoing lumbar spinal fusion surgery

1. The association between advanced glycation end products derived from confocal fluorescence microscopy of bone biopsies and dermal ultrasound echogenicity in patients undergoing lumbar spinal fusion surgery

1. The association between advanced glycation end products derived from confocal fluorescence microscopy of bone biopsies and dermal ultrasound echogenicity in patients undergoing lumbar spinal fusion surgery

Oleuropein-Enriched Extract From Olive Mill Leaves by Homogenizer-Assisted Extraction and Its Antioxidant and Antiglycating Activities.

Olive oil consumption has increased in the last two decades and consequently, its wastes have increased, which generates a tremendous environmental impact. Among the by-products are the olive mill leaves, which are easier and inexpensive to treat than other olive by-products. However, little research has been done on their chemical composition and potential bioactivity. Hence, in this study, olive mill leaves were used to obtain Oleuropein-Enriched Extracts (OLEU-EE) using Conventional Extraction, Ultrasound-Assisted Extraction, and Homogenization-Assisted Extraction. These three techniques were evaluated using a Factorial Design to determine the parameters to obtain an OLEU-EE with high contents of Total Phenolic Compounds (TPC), Antioxidant Activity (AA), and Oleuropein concentration (OLEU). From the results, the Homogenizer-Assisted Extraction (HAE) technique was selected at 18,000 rpm, solid:liquid ratio 1:10, and 30 s of homogenization with 70% ethanol, due to its high TPC (5,196 mg GA/100 g), AA (57,867 μmol of TE/100 g), and OLEU (4,345 mg of OLEU/100 g). In addition, the antiglycating effect of OLEU-EE on the levels of (1) fluorescent Advanced Glycation End Products (AGEs) were IC50 of 0.1899 and 0.1697 mg/mL for 1λEXC 325/λEM 440 and 2λEXC 389/λEM 443, respectively; (2) protein oxidative damage markers such as dityrosine (DiTyr), N-formylkynurenine (N-formyl Kyn), and kynurenine (Kyn) were IC50 of 0.1852, 0.2044, and 0.1720 mg/mL, respectively. In conclusion, OLEU-EE from olive mill leaves has different capacities to inhibit AGEs evidenced by the IC50 of fluorescent AGEs and protein oxidation products, together with the scavenging free radical evidenced by the concentration of Trolox Equivalent. Therefore, OLEU-EE could be potential functional ingredients that prevent oxidative damage caused by free radicals and AGEs accumulation.

Adolescent obesity incurs adult skeletal deficits in murine induced obesity model.

Adolescent obesity has risen dramatically in the last few decades. While adult obesity may be osteoprotective, the effects of obesity during adolescence, which is a period of massive bone accrual, are not clear. We used a murine model of induced adolescent obesity to examine the structural, mechanical, and compositional differences between obese and healthy weight bone in 16-week-old female C57Bl6 mice. We also examined the effects of a return to normal weight after skeletal maturity (24 weeks old). We found obese adolescent bone exhibited decreased trabecular bone volume, increased cortical diameter, increased ultimate stress, and increased brittleness (decreased plastic energy to fracture), similar to an aging phenotype. The trabecular bone deficits remained after return to normal weight after skeletal maturity. However, after returning to normal diet, there was no difference in ultimate stress nor plastic energy to fracture between groups as the normal diet group increased ultimate stress and brittleness. Interestingly, compositional changes appeared in the former high-fat diet mice after skeletal maturity with a lower mineral to matrix ratio compared to normal diet mice. In addition there was a trend toward increased fluorescent advanced glycation endproducts in the former high-fat diet mice compared to normal diet mice but this did not reach significance (p < 0.05) due to the large variability. The skeletal consequences of adolescent obesity may have lasting implications for the adult skeleton even after return to normal weight. Given the rates of adolescent obesity, skeletal health should be a concern.