A review on methods of estimation of advanced glycation end products

| Posted in: Science

Journal name: World Journal of Pharmaceutical Research
Original article title: A review on methods of estimation of advanced glycation end products
The WJPR includes peer-reviewed publications such as scientific research papers, reports, review articles, company news, thesis reports and case studies in areas of Biology, Pharmaceutical industries and Chemical technology while incorporating ancient fields of knowledge such combining Ayurveda with scientific data.
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Original source:

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Author:

Swathi Putta and Eswar Kumar Kilari


World Journal of Pharmaceutical Research:

(An ISO 9001:2015 Certified International Journal)

Full text available for: A review on methods of estimation of advanced glycation end products

Source type: An International Peer Reviewed Journal for Pharmaceutical and Medical and Scientific Research


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Summary of article contents:

Introduction

Diabetes mellitus poses significant public health challenges, with an estimated 220 million people afflicted worldwide by 2020. Chronic diabetes leads to a range of complications primarily through the formation of advanced glycation end products (AGEs). AGEs accumulate in long-lived proteins, driving inflammation and thickening of basement membranes. Notable complications associated with high levels of AGEs include retinopathy, neuropathy, nephropathy, and atherosclerosis. The study focuses on the various techniques utilized to detect AGEs, highlighting their relevance in understanding diabetes-related complications.

Importance of Advanced Glycation End Products (AGEs)

AGRs are formed through the non-enzymatic glycation of proteins and lipids, a process enhanced by hyperglycemia and oxidative stress frequently observed in diabetic patients. Commonly studied AGE markers include N-carboxymethyllysine (CML), pentosidine, and methylglyoxal (MG). The quantification of these products provides insight into a patient's past metabolic control and glucose exposure, offering a dual perspective. Early glycation product measurements help estimate glucose levels, while the assessment of intermediate and late glycation products aids in understanding tissue modifications and associated pathology.

Techniques for Measuring AGEs: Spectrofluorimetry

A prominent method for measuring AGEs involves spectrofluorimetry, which analyzes the fluorescence intensity of glycated materials. In the procedure, test samples mixed with trichloroacetic acid (TCA) undergo centrifugation, followed by measurement of fluorescence at specific wavelengths using a spectrofluorometer. Assays involving bovine serum albumin (BSA) demonstrate how in vitro glycation can be assessed, including protocols for incubating BSA with glucose or fructose to evaluate the inhibitory effects of test compounds on AGE formation. This technique is essential in gauging the presence of AGEs in various biological samples.

Techniques for Measuring AGEs: HPLC and ELISA

High-Performance Liquid Chromatography (HPLC) and Enzyme-Linked Immunosorbent Assay (ELISA) also serve critical roles in AGE detection. HPLC enables size-selective separation of serum and urine samples, allowing the identification of glycation products based on retention times. The methodology utilizes specific columns and elution conditions tailored to separate AGEs from other components. Conversely, ELISA offers a targeted approach for quantifying specific AGE markers like CML. This technique leverages antibodies against AGEs, providing quantitative results based on the absorbance of the colored reaction product. Both methods contribute significantly to the understanding of AGEs in diabetic pathology.

Conclusion

Regular measurement of glycation products is vital for the metabolic monitoring of diabetic patients, offering insights into their risk for developing chronic complications. Advancements in detection techniques, including spectrofluorimetry, HPLC, and ELISA, facilitate meticulous tracking of AGEs, thereby enhancing our understanding of their role in the pathogenesis of diabetes complications. Armed with knowledge from these methodologies, healthcare professionals can better assess and manage the implications of chronic hyperglycemia, providing a strong basis for effective diabetes care strategies.

FAQ section (important questions/answers):

What are advanced glycation end products (AGEs) and their significance?

AGEs are harmful compounds formed when sugars react with proteins. They accumulate in tissues, leading to complications like retinopathy, neuropathy, and nephropathy, particularly in diabetes.

Why is measuring AGEs important in diabetes patients?

Measuring AGEs helps assess metabolic control and the extent of tissue damage due to chronic exposure to high glucose levels, indicating the risk of developing complications.

What techniques are used to detect AGEs in research?

Common techniques for detecting AGEs include Spectrofluorimetry, ELISA, HPLC, and Mass Spectrometry. Each method has distinct protocols and sensitivities for AGE measurement.

How does the Spectrofluorimetric method for AGEs work?

In Spectrofluorimetry, AGE samples are mixed with TCA, centrifuged, and fluorescence is measured at specific wavelengths, allowing quantification of AGEs in various biological samples.

What role do AGEs play in diabetic complications?

AGs contribute to diabetes complications by inducing inflammation, crosslinking proteins, and thickening membranes, exacerbating conditions like atherosclerosis and kidney damage.

How can dietary intake affect AGE levels in the body?

Dietary intake, particularly the consumption of high-sugar and processed foods, can increase circulating AGEs, intensifying oxidative stress and raising the risk of diabetes complications.

Glossary definitions and references:

Scientific and Ayurvedic Glossary list for “A review on methods of estimation of advanced glycation end products”. This list explains important keywords that occur in this article and links it to the glossary for a better understanding of that concept in the context of Ayurveda and other topics.

1) Water:
Water is a vital component in biological reactions and physiological processes, including the formation of advanced glycation end products (AGEs). It serves as a solvent for biochemical substances and is essential for maintaining normal cellular functions. The properties of water also influence the solubility and reactivity of various compounds in pharmaceutical studies.

2) Measurement:
Measurement is a crucial aspect of scientific research, particularly in the context of quantifying advanced glycation end products (AGEs). Accurate measurement methods such as HPLC, ELISA, and spectrofluorimetry enable researchers to analyze the levels of AGEs in biological samples, facilitating the understanding of their correlation with diabetes and related complications.

3) Andhra (Amdhra):
Andhra refers to the state in India where the authors of the study are based (Andhra University, Visakhapatanm, Andhra Pradesh). This geographic context is significant as it highlights the regional scope of the research on advanced glycation end products and their relevance to local health concerns related to diabetes.

4) Sugar:
Sugar, in the form of glucose and other carbohydrates, is critical in the formation of advanced glycation end products (AGEs) through non-enzymatic glycation reactions. Understanding sugar's role is vital because elevated glucose levels in diabetic patients lead to increased AGE formation, contributing to chronic complications such as retinopathy and neuropathy.

5) Activity:
Activity in this context refers to the biological and chemical activities related to substances being studied, particularly the inhibitory effects that certain compounds may have on AGE formation. Assessing the activity of potential AGE inhibitors helps identify therapeutic angles for managing diabetes and mitigating its complications.

6) Science (Scientific):
Scientific pertains to the rigorous methodologies employed in research, particularly in studying advanced glycation end products (AGEs) and their impact on diabetes. Scientific approaches underpin the experimental design, data analysis, and interpretation of results, ensuring that findings are credible and can contribute to advancements in medical knowledge.

7) Kumar:
Kumar, referring to one of the authors, Eswar Kumar Kilari, signifies the contribution of individual researchers in the scientific field. Authors play a critical role in the dissemination of knowledge, guiding investigations into advanced glycation end products and their implications for diabetes and pharmaceutical science.

8) Miṇi (Mini):
In this context, 'mini' can refer to the mini gel electrophoresis technique used in the western-blotting detection methods for analyzing AGE-modified proteins. This technique is essential for the resolution of protein samples, enabling the visualization of specific glycation modifications that contribute to the understanding of diabetic complications.

9) Hand:
Hand can relate to the manual techniques employed in laboratory processes, including sample preparation, handling of test materials, and interactions during measurements. Proper hand techniques are vital to ensure accuracy and reproducibility in experiments measuring advanced glycation end products and understanding their implications for diabetes.

10) Accumulation (Accumulating, Accumulate):
Accumulate refers to the gradual increase in the concentration of advanced glycation end products (AGEs) within biological systems, predominantly in individuals with diabetes. This accumulation serves as a biomarker for monitoring metabolic dysfunction and can help assess the risk of developing chronic complications associated with diabetes.

11) Inflammation:
Inflammation is a biological response to harmful stimuli, such as advanced glycation end products (AGEs), which can trigger inflammatory pathways in diabetic patients. Understanding the link between AGEs and inflammation is crucial for comprehending how these compounds exacerbate diabetes-related complications and for developing therapeutic strategies.

12) Substance:
Substance refers to any specific compound or material, such as advanced glycation end products, which are of interest in the study. The identification, quantification, and understanding of such substances are critical for determining their role in diabetes and the potential for therapeutic interventions.

13) Resolving:
Resolving in laboratory techniques can refer to the separation and identification of complex mixtures of substances, such as proteins modified by advanced glycation end products. Efficient resolving methods, like mass spectrometry and HPLC, are essential for analyzing these compounds and understanding their biological implications in diabetes.

14) Relative:
Relative often pertains to the comparison between different measurements or conditions in scientific research. In the context of AGEs, relative quantification may be used to compare levels of these compounds in different biological samples, thus revealing associations with disease states or progression in diabetic patients.

15) Gelatin:
Gelatin may refer to a substance used in various laboratory protocols, potentially as a blocking agent in ELISA techniques. Its function is to prevent non-specific binding during assays measuring advanced glycation end products, enhancing the accuracy of the results obtained from such sensitive measurements.

16) Putta:
Putta refers to Swathi Putta, one of the authors of the study, representing the collaborative nature of scientific research. The contributions of researchers like Putta are essential in advancing knowledge about advanced glycation end products and their relevance in diabetes management and complications.

17) Blood:
Blood is a critical biological fluid for assessing advanced glycation end products, as it serves as a transport medium for these compounds in the circulation. Measuring AGEs in blood helps researchers understand their systemic levels and correlate them with diabetes complications, enhancing disease management strategies.

18) Study (Studying):
Study refers to the systematic investigation into advanced glycation end products, their formation, measurement methods, and their implications for diabetes. It embodies the essence of scientific inquiry aimed at expanding knowledge and providing insights that may influence clinical practices and therapeutic approaches.

19) Line:
Line can represent the rationale or reasoning drawn from data collected during experiments. In research, establishing a basis for different measurements of advanced glycation end products often depends on a logical line of thought that connects observations with underlying biological mechanisms affecting diabetes progression.

20) Gold (Golden):
Gold may refer to specific advanced glycation end products, like GOLD and pentosidine, which are analyzed within the context of diabetes. Their identification and quantification can provide valuable insights into the pathophysiology of diabetic complications and serve as biomarkers for disease monitoring and intervention efficacy.

Other Science Concepts:

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Discover the significance of concepts within the article: ‘A review on methods of estimation of advanced glycation end products’. Further sources in the context of Science might help you critically compare this page with similair documents:

Clinical studies, Mass spectrometry, Chronic diabetes, HPLC Method, Advanced glycation end products.

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