Dysregulation of Receptor for Advanced Glycation End Products (RAGE) Expression as a Biomarker of Keratoconus

Background. Because of the implications of Receptor for Advanced Glycation End Products (RAGE) in keratoconus (KC), we describe a differential expression of RAGE transcripts and proteins in corneal tissues and tears of KC and healthy patients. Methods. Using a case-controlled study, corneal epitheliums and tears of KC and healthy subjects were obtained during corneal collagen cross-linking and photorefractive keratectomy (PKR) and during usual consultations. Quantitative reverse transcription (RT-qPCR) and Western-Blot were performed to analyze RAGE transcripts and proteins’ expression in corneal tissues and tears. Results. One hundred and six patients were included in this study. The characteristics of the patients were as follows: 56 KC (25 corneal epithelium and 31 tears) and 50 control subjects (25 corneal epithelium and 25 tears). Transcripts of RAGE, HMGB1, and S100 family ligands were quantified by RT-qPCR, identifying a significantly higher expression of RAGE and HMGB1 in the healthy group than in the KC group ( p = 0.03 and 0.04, respectively). Western Blot showed a significantly higher fl-RAGE expression in KC corneal epithelium than control ( p < 0.001 ) and lower s-RAGE expression in KC tears than control ( p = 0.04 ). Conclusions. Linked with the inflammatory process occurring in KC pathophysiology, we propose for the first time that the RAGE expression (total and truncated forms of receptor and ligands) in KC corneal tissues and tear samples provides viable biomarkers.

Anti-high-mobility group box-1 (HMGB1) mediates the apoptosis of alveolar epithelial cells (AEC) by receptor of advanced glycation end-products (RAGE)/c-Jun N-terminal kinase (JNK) pathway in the rats of crush injuries

Objectives The lung injury is often secondary to severe trauma. In the model of crush syndrome, there may be secondary lung injury. We hypothesize that high-mobility group box 1 (HMGB1), released from muscle tissue, mediates the apoptosis of alveolar epithelial cells (AEC) via HMGB1/Receptor of advanced glycation end-products (RAGE)/c-Jun N-terminal kinase (JNK) pathway. The study aimed to investigate how HMGB1 mediated the apoptosis of AEC in the rat model. Methods Seventy-five SD male rats were randomly divided into five groups: CS, CS + vehicle, CS + Ethyl pyruvate (EP), CS + FPS-ZM1 group, and CS + SP600125 groups. When the rats CS model were completed after 24 h, the rats were sacrificed. We collected the serum and the whole lung tissues. Inflammatory cytokines were measured in serum samples. Western blot and RT-qPCR were used to quantify the protein and mRNA. Lastly, apoptotic cells were detected by TUNEL. We used SPSS 25.0 for statistical analyses. Results Nine rats died during the experiments. Dead rats were excluded from further analysis. Compared to the CS group, levels of HMGB1 and inflammatory cytokines in serum were downregulated in CS + EP, CS + FPS-ZM1, and CS + SP600125 groups. Western blot and RT-qPCR analysis revealed a significant downregulation of HMGB1, RAGE, and phosphorylated-JNK in CS + EP, CS + FPS-ZM1, and CS + SP600125 groups, compared with the CS groups, excluding total-JNK mRNA. Apoptosis of AEC was used TUNEL to assess. We found the TUNEL-positive cells were downregulated in CS + EP, CS + FPS-ZM1, and CS + SP600125 groups. Conclusion The remote lung injury begins early after crush injuries. The HMGB1/RAGE/JNK signaling axis is an attractive target to abrogate the apoptosis of AEC after crush injuries.

In Vitro Methodologies to Study the Role of Advanced Glycation End Products (AGEs) in Neurodegeneration

Advanced glycation end products (AGEs) can be present in food or be endogenously produced in biological systems. Their formation has been associated with chronic neurodegenerative diseases such as Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, and amyotrophic lateral sclerosis. The implication of AGEs in neurodegeneration is related to their ability to bind to AGE-specific receptors and the ability of their precursors to induce the so-called “dicarbonyl stress”, resulting in cross-linking and protein damage. However, the mode of action underlying their role in neurodegeneration remains unclear. While some research has been carried out in observational clinical studies, further in vitro studies may help elucidate these underlying modes of action. This review presents and discusses in vitro methodologies used in research on the potential role of AGEs in neuroinflammation and neurodegeneration. The overview reveals the main concepts linking AGEs to neurodegeneration, the current findings, and the available and advisable in vitro models to study their role. Moreover, the major questions regarding the role of AGEs in neurodegenerative diseases and the challenges and discrepancies in the research field are discussed.

Receptor Mediated Effects of Advanced Glycation End Products (AGEs) on Innate and Adaptative Immunity: Relevance for Food Allergy

As of late, evidence has been emerging that the Maillard reaction (MR, also referred to as glycation) affects the structure and function of food proteins. MR induces the conformational and chemical modification of food proteins, not only on the level of IgG/IgE recognition, but also by increasing the interaction and recognition of these modified proteins by antigen-presenting cells (APCs). This affects their biological properties, including digestibility, bioavailability, immunogenicity, and ultimately their allergenicity. APCs possess various receptors that recognize glycation structures, which include receptor for advanced glycation end products (RAGE), scavenger receptors (SRs), galectin-3 and CD36. Through these receptors, glycation structures may influence the recognition, uptake and antigen-processing of food allergens by dendritic cells (DCs) and monocytes. This may lead to enhanced cytokine production and maturation of DCs, and may also induce adaptive immune responses to the antigens/allergens as a result of antigen uptake, processing and presentation to T cells. Here, we aim to review the current literature on the immunogenicity of AGEs originating from food (exogenous or dietary AGEs) in relation to AGEs that are formed within the body (endogenous AGEs), their interactions with receptors present on immune cells, and their effects on the activation of the innate as well as the adaptive immune system. Finally, we review the clinical relevance of AGEs in food allergies.

Intracellular Toxic Advanced Glycation End-Products in 1.4E7 Cell Line Induce Death with Reduction of Microtubule-Associated Protein 1 Light Chain 3 and p62

Background: The death of pancreatic islet β-cells (β-cells), which are the insulin-producing cells, promote the pathology in both Type 1 and Type 2 diabetes mellitus (DM) (T1DM and T2DM), and they are protected by autophagy which is one of the mechanisms of cell survival. Recently, that some advanced glycation end-products (AGEs), such as methylglyoxial-derived AGEs and Nε-carboxymethyllysine, induced the death of β-cells were revealed. In contrast, we had reported AGEs derived from glyceraldehyde (GA, the metabolism intermediate of glucose and fructose) are considered to be toxic AGEs (TAGE) due to their cytotoxicity and role in the pathogenesis of T2DM. More, serum levels of TAGE are elevated in patients with T1 and T2DM, where they exert cytotoxicity. Aim: We researched the cytotoxicity of intracellular and extracellular TAGE in β-cells and the possibility that intracellular TAGE were associated with autophagy. Methods: 1.4E7 cells (a human β-cell line) were treated with GA, and analyzed viability, quantity of TAGE, microtubule-associated protein 1 light chain 3 (LC3)-I, LC3-II, and p62. We also examined the viability of 1.4E7 cells treated with TAGE-modified bovine serum albumin, a model of TAGE in the blood. Results: Intracellular TAGE induced death of 1.4E7 cells, decrease of LC3-I, LC3-II, and p62. Extracellular TAGE didn’t show cytotoxicity in the physiological concentration. Conclusion: Intracellular TAGE induced death of β-cells more strongly than extracellular TAGE, and may suppress autophagy via reduction of LC3-I, LC3-II, and p62 to inhibit the degradation of them.

Microbial protein metabolism in the monogastric gastrointestinal tract: a review

Dietary and endogenous protein that become available for the microbiota in the hindgut can be metabolized via different routes. They can become building blocks for the microbial cells or enter different catabolic pathways. Protein degradation via fermentation pathways is seen as a non-preferred route as it results in the formation and release of metabolites that can interfere with biological systems in the host and can have deleterious outcomes. Reducing protein fermentation and guiding the metabolism towards less toxic end-products might be possible targets for improving host health. To do so, more knowledge on factors manipulating the process of microbial protein metabolism, including on substrate availability, microbial composition and segmental differences in the hindgut, is required.

Pyroelectrometallurgical processing of bismuth-containing oxides

In this work, we substantiate and develop a general pyroelectrometallurgical technology for processing bismuth dross and oxides (the intermediate products of lead bullion refining by the Betterton-Kroll process) to obtain crude bismuth. The research focuses on bismuth dross (3–5% Bi; 80–85% Pb) remelted at 500–600°С in the presence of NaNO3 and NaOH, as well as the obtained alkaline melt (bismuth oxides, 1–5% Bi; 60–70% Pb). The conducted experiments allowed us to determine optimal parameters of the main steps of processing bismuth oxide, as well as the characteristics of obtained products. Reduction smelting of bismuth oxides at 1150°C (with the addition of sodium carbonate, quartz and fine coke in the amount of 66, 25 and 5% of bismuth oxides mass, respectively) is proposed, leading to bismuth lead formation. Its decoppering is carried out at 350–600°C with 2.0% sulfur (by its weight), added to the melt. We propose to carry out the alkaline treatment of the decoppered Pb-Bi alloy at 500oC in contact with sodium hydroxide, sodium nitrate and sodium chloride, taken in amounts up to 10.2, 8.3 and 1.4% by weight of bismuth lead, respectively. Subsequent electrolysis comprises electrolytic processing of purified Pb-Bi alloy ingots at 550oC. The electrolyte consists of a melt with the following composition, %: NaCl – 7, KCl – 35, PbCl2 – 18 and ZnCl2 – 40. As a result, two end products were obtained by the proposed bismuth oxide processing. The anodic product at the second stage of electrolysis, crude bismuth (yielded 1.1% by the weight of oxides) contains 93.62% Bi and 4.14% Pb, extraction from oxides amounts to 19.0% Bi and 0.1% Pb. About 1.2% Bi and 9.1% Pb of their initial content in the oxides are transferred to the cathodic product containing 0.033% Bi and 97.83% Pb (the yield equalled 5.1% of the oxides).

Biochemical analyses for dental age estimation: a review

Background For various legal and forensic scenarios, establishing an individual’s age, both living and dead, plays a crucial role. Various morphological, radiographic, and molecular methods can be used for age estimation. In children and adolescents, age estimation is based on the established developmental stages. However, in adults, where the development ceases into maturation, the degenerative changes play a role in determining the age. Main body of the abstract In the natural aging process, several molecular changes occur most commonly in the long-living proteins and hard tissues like the teeth and bone. These molecular changes gradually lead to alterations in several organs and organ systems, which can be quantified and correlated with age, including aspartic acid racemization, collagen crosslinks, advanced glycation-end products, and mitochondrial DNA mutations. Short conclusion Among the above methods, the racemization of aspartic acid can be considered as the most precise method. The main advantage of using aspartic acid racemization is that the sample can be collected from tissues (teeth) protected from various environmental and nutritional factors. If all the confounding factors are stable, the utilization of advanced glycation-end products can also be considered valuable. Environmental factors like lead accumulations may also help determine the age. However, further studies need to be conducted, focusing on providing a more standardized method. This review provides a concise summary of the biochemical techniques that can be used for estimation of age.