This document discusses genetic markers related to non-alcoholic fatty liver disease (NAFLD). The study aimed to identify genetic markers that could help develop non-invasive diagnostic methods for NAFLD. The study identified 6 potentially associated genes, including the first association between SLC2A1 and NAFLD. Investigation of these genes in liver biopsies found they are regulated in NAFLD, and regulation increases with disease advancement, suggesting they could serve as markers for diagnosing NAFLD development phases.
Biochemical Study on Endothelial Nitric Oxide Gene Polymorphism in Fatty Live...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Talk : Nash in children by Dr. Pramod MistrySanjeev Kumar
This document discusses non-alcoholic fatty liver disease (NAFLD) in children, including:
1) NAFLD prevalence in children is difficult to determine due to selection bias and lack of definitive diagnoses, though it is likely higher in India than Western countries.
2) Risk factors for pediatric NAFLD include genes associated with inefficient lipid processing combined with an obesogenic environment of excessive calories and sedentary lifestyle.
3) Evaluation of suspected NAFLD includes blood tests, imaging, and sometimes liver biopsy, though biopsy remains the gold standard for diagnosis and assessing severity.
Fatty liver, or hepatic steatosis, refers to excess triglyceride accumulation in liver cells. This can be caused by increased fatty acid mobilization from adipose tissue, excess fatty acid synthesis in the liver, or reduced removal of fat from the liver. Common causes include obesity, uncontrolled diabetes, high-fat diet, alcoholism, and deficiencies in lipotropic factors like choline, betaine, and inositol. Untreated, fatty liver can progress to non-alcoholic steatohepatitis and fibrosis or cirrhosis of the liver.
Fatty liver disease, also known as hepatic steatosis, refers to a reversible condition where triglyceride fat accumulates in the liver cells. It is commonly associated with alcohol use, metabolic syndrome, diabetes, hypertension, obesity or dyslipidemia. Both alcoholic and non-alcoholic fatty liver disease show micro and macro vesicular fatty changes at different stages, making it difficult to distinguish between the two. The treatment of fatty liver depends on the underlying cause, and may involve lifestyle changes like diet, exercise and reducing alcohol intake or controlling blood sugar.
1) Diabetes mellitus is a disorder of carbohydrate metabolism caused by insufficient insulin production or ineffective insulin, resulting in high blood glucose levels.
2) There are several types of diabetes classified by their causes, including type 1 caused by autoimmune destruction of insulin-producing beta cells, and type 2 caused by insulin resistance and relative insulin deficiency.
3) Key organs involved in regulating blood glucose include the pancreas, which produces insulin and glucagon, and the liver, which stores glucose as glycogen and releases it as needed.
Effects of chicory (Cichorium intybus L.) on nonalcoholic fatty liver diseaseLucyPi1
Abstract There is a dramatic increase in the prevalence of nonalcoholic fatty liver disease, which is slowly turning into a pandemic as well as a major challenge across the world. Nonalcoholic fatty liver disease is described as a range of liver conditions such as fat accumulation, hepatic steatosis, or end-stage liver disease. Patients with nonalcoholic fatty liver disease are asymptomatic and their mortality is higher than people without nonalcoholic fatty liver disease. The pathogenesis of nonalcoholic fatty liver disease has not been clearly determined yet. The “two hits” hypothesis is designed to explain the pathogenesis of nonalcoholic fatty liver disease. Dyslipidemia, oxidative stress, insulin resistance, obesity, and chronic inflammation are some of the morbidities involved in the progression of nonalcoholic fatty liver disease. Chicory (Cichorium intybus L.) is an herbaceous perennial, known as chicory. Chicory contains various compounds, such as vitamins, sonchuside A, caffeic acid derivatives, fructo-oligosaccharides, chlorogenic acid, magnolialide, polysaccharides, coumarins, phenolic acids, terpenoids, flavonoids, polyphenol, cichoriosides, ixerisosides, eudesmanolides, inulin, bitter sesquiterpene lactones, and alkaloids. Current research has revealed that chicory supplementation might be effective in the treatment of nonalcoholic fatty liver disease. The anti-inflammatory, antihepatotoxic, antihyperlipidemic, antidiabetic, antihyperglycemic, and antioxidant properties of chicory provide plausible mechanisms by which chicory may affect the various steps of disease progression and severity. Existing studies have shown that chicory supplementation has beneficial effects on nonalcoholic fatty liver disease, but the existence of only one human study and possible side effects of chicory necessitate further studies.
Hiperuricemia diabetes y sindrome metabolico2Residentes1hun
Insulin resistance plays a potentially key role in the causal relationship between metabolic syndrome, type 2 diabetes and hyperuricemia. Evidence suggests that insulin resistance may be a mediator linking obesity, inflammation, and hyperuricemia. Furthermore, hyperuricemia and insulin resistance likely share a bidirectional causal effect, as insulin resistance can lead to hyperuricemia through impaired kidney function, while hyperinsulinemia can also decrease uric acid excretion and lead to hyperuricemia.
This document discusses metabolic syndrome, which is a combination of medical disorders that increase the risk of cardiovascular disease and diabetes when occurring together. It affects about 20% of the Malaysian population. The core components include hypertension, high triglycerides, low HDL cholesterol, obesity, and impaired glucose tolerance. There are different criteria for diagnosing metabolic syndrome, but central to all definitions is insulin resistance. If left untreated, metabolic syndrome can increase the risk of serious health conditions such as heart disease, stroke, and type 2 diabetes.
Biochemical Study on Endothelial Nitric Oxide Gene Polymorphism in Fatty Live...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
Talk : Nash in children by Dr. Pramod MistrySanjeev Kumar
This document discusses non-alcoholic fatty liver disease (NAFLD) in children, including:
1) NAFLD prevalence in children is difficult to determine due to selection bias and lack of definitive diagnoses, though it is likely higher in India than Western countries.
2) Risk factors for pediatric NAFLD include genes associated with inefficient lipid processing combined with an obesogenic environment of excessive calories and sedentary lifestyle.
3) Evaluation of suspected NAFLD includes blood tests, imaging, and sometimes liver biopsy, though biopsy remains the gold standard for diagnosis and assessing severity.
Fatty liver, or hepatic steatosis, refers to excess triglyceride accumulation in liver cells. This can be caused by increased fatty acid mobilization from adipose tissue, excess fatty acid synthesis in the liver, or reduced removal of fat from the liver. Common causes include obesity, uncontrolled diabetes, high-fat diet, alcoholism, and deficiencies in lipotropic factors like choline, betaine, and inositol. Untreated, fatty liver can progress to non-alcoholic steatohepatitis and fibrosis or cirrhosis of the liver.
Fatty liver disease, also known as hepatic steatosis, refers to a reversible condition where triglyceride fat accumulates in the liver cells. It is commonly associated with alcohol use, metabolic syndrome, diabetes, hypertension, obesity or dyslipidemia. Both alcoholic and non-alcoholic fatty liver disease show micro and macro vesicular fatty changes at different stages, making it difficult to distinguish between the two. The treatment of fatty liver depends on the underlying cause, and may involve lifestyle changes like diet, exercise and reducing alcohol intake or controlling blood sugar.
1) Diabetes mellitus is a disorder of carbohydrate metabolism caused by insufficient insulin production or ineffective insulin, resulting in high blood glucose levels.
2) There are several types of diabetes classified by their causes, including type 1 caused by autoimmune destruction of insulin-producing beta cells, and type 2 caused by insulin resistance and relative insulin deficiency.
3) Key organs involved in regulating blood glucose include the pancreas, which produces insulin and glucagon, and the liver, which stores glucose as glycogen and releases it as needed.
Effects of chicory (Cichorium intybus L.) on nonalcoholic fatty liver diseaseLucyPi1
Abstract There is a dramatic increase in the prevalence of nonalcoholic fatty liver disease, which is slowly turning into a pandemic as well as a major challenge across the world. Nonalcoholic fatty liver disease is described as a range of liver conditions such as fat accumulation, hepatic steatosis, or end-stage liver disease. Patients with nonalcoholic fatty liver disease are asymptomatic and their mortality is higher than people without nonalcoholic fatty liver disease. The pathogenesis of nonalcoholic fatty liver disease has not been clearly determined yet. The “two hits” hypothesis is designed to explain the pathogenesis of nonalcoholic fatty liver disease. Dyslipidemia, oxidative stress, insulin resistance, obesity, and chronic inflammation are some of the morbidities involved in the progression of nonalcoholic fatty liver disease. Chicory (Cichorium intybus L.) is an herbaceous perennial, known as chicory. Chicory contains various compounds, such as vitamins, sonchuside A, caffeic acid derivatives, fructo-oligosaccharides, chlorogenic acid, magnolialide, polysaccharides, coumarins, phenolic acids, terpenoids, flavonoids, polyphenol, cichoriosides, ixerisosides, eudesmanolides, inulin, bitter sesquiterpene lactones, and alkaloids. Current research has revealed that chicory supplementation might be effective in the treatment of nonalcoholic fatty liver disease. The anti-inflammatory, antihepatotoxic, antihyperlipidemic, antidiabetic, antihyperglycemic, and antioxidant properties of chicory provide plausible mechanisms by which chicory may affect the various steps of disease progression and severity. Existing studies have shown that chicory supplementation has beneficial effects on nonalcoholic fatty liver disease, but the existence of only one human study and possible side effects of chicory necessitate further studies.
Hiperuricemia diabetes y sindrome metabolico2Residentes1hun
Insulin resistance plays a potentially key role in the causal relationship between metabolic syndrome, type 2 diabetes and hyperuricemia. Evidence suggests that insulin resistance may be a mediator linking obesity, inflammation, and hyperuricemia. Furthermore, hyperuricemia and insulin resistance likely share a bidirectional causal effect, as insulin resistance can lead to hyperuricemia through impaired kidney function, while hyperinsulinemia can also decrease uric acid excretion and lead to hyperuricemia.
This document discusses metabolic syndrome, which is a combination of medical disorders that increase the risk of cardiovascular disease and diabetes when occurring together. It affects about 20% of the Malaysian population. The core components include hypertension, high triglycerides, low HDL cholesterol, obesity, and impaired glucose tolerance. There are different criteria for diagnosing metabolic syndrome, but central to all definitions is insulin resistance. If left untreated, metabolic syndrome can increase the risk of serious health conditions such as heart disease, stroke, and type 2 diabetes.
1. Steatosis, or fatty liver, is common and usually benign but can progress to steatohepatitis if associated with conditions like obesity, diabetes, or alcohol use.
2. Alcohol is a leading cause of steatohepatitis and the most common type of liver disease in Western nations, due to the toxic byproducts produced when the liver metabolizes alcohol.
3. The pathology of alcohol-related liver disease ranges from reversible fatty liver to fibrosis, cirrhosis, and even liver cancer. Management involves cessation of alcohol and treatment of complications.
This document provides an overview of metabolic pathways and inborn errors of metabolism. It discusses the major metabolic pathways for carbohydrates, proteins, and lipids and how genetic defects can disrupt these pathways. It also describes the three main types of inborn errors of metabolism and provides examples. The document then goes into more detail about carbohydrate metabolism, including glycolysis, pyruvate metabolism, and the tricarboxylic acid cycle. It also discusses glycogen metabolism, protein metabolism, fat metabolism, and approaches to investigating metabolic cases.
This document provides an overview of diabetes mellitus, including its classification, pathophysiology, clinical features, investigations, diagnostic criteria, and management. It discusses the different types of diabetes, risk factors, characteristics, and laboratory findings. Type 1 diabetes results from beta cell destruction leading to insulin deficiency, while type 2 involves insulin resistance with relative insulin deficiency. Gestational diabetes occurs during pregnancy.
This document reviews the cellular mechanisms and clinical evidence regarding curcumin's effects on oxidative stress-related liver diseases. It finds that curcumin exerts remarkable protective and therapeutic effects through various mechanisms, including suppressing proinflammatory cytokines and lipid peroxidation, activating protective signaling pathways, and ameliorating cellular responses to oxidative stress like increasing antioxidant enzyme expression. Curcumin itself acts as a free radical scavenger against various reactive oxygen species due to its phenolic, β-diketone, and methoxy functional groups. Further clinical studies are still needed to better understand curcumin's structure-activity relationships and mechanisms in oxidative liver diseases.
Diabetes mellitus, its types and compicationsMohit Adhikary
This document discusses types and complications of diabetes mellitus. It begins with an outline that defines diabetes and classifies diabetes types and complications as acute or chronic. It then discusses the various types of diabetes in more detail, including type 1 diabetes pathogenesis and genetic and environmental risk factors. Type 2 diabetes risk factors and pathophysiology involving insulin resistance and secretion are covered. Other specific rare genetic types are defined. The document concludes by examining acute complications like diabetic ketoacidosis and chronic complications involving microvascular and macrovascular involvement, as well as theories around how hyperglycemia may lead to these complications. Glycemic control studies proving the benefits of control are also summarized.
Diabetes mellitus (DM) refers to a group of common metabolic disorders that share the phenotype of hyperglycemia, due to defect in insulin secretion, insulin action or both .
Beta oxidation defects are inborn errors of metabolism that result in the failure of fatty acid oxidation. There are several types depending on the length of the fatty acid involved: very long chain, long chain, medium chain, or short chain. The main pathways affected are carnitine transport, beta oxidation, and ketogenesis. Presentations range from hypoglycemia in infants to muscle weakness or exercise intolerance in older children and adults. Diagnosis involves testing for intermediary metabolites, acylcarnitines, and enzyme activities. Treatment focuses on avoiding fasting and providing rapid glucose treatment during acute illnesses.
This document discusses diabetes mellitus (DM), including:
1) DM is a chronic disease characterized by high blood sugar levels due to either the pancreas not producing enough insulin or the body not properly using the insulin produced.
2) There are two main types of DM - type 1 is usually diagnosed in childhood/early adulthood and results from autoimmune destruction of insulin-producing beta cells, while type 2 accounts for 80% of cases and involves insulin resistance and relative lack of insulin.
3) Uncontrolled DM can lead to serious damage of various organs and body systems, and was responsible for an estimated 1.5 million deaths in 2012. Lifestyle changes such as healthy diet, exercise and weight control can help
The document discusses calcium, phosphate, and magnesium content and distribution in the human body. It also describes the physiological functions of calcium, including cellular functions like cell growth and division, and extracellular functions like mineralization and blood clotting. Calcium signaling is discussed, where calcium binds to and activates proteins like calmodulin and calcium ATPase pumps.
1. A 65-year-old male presented with fever, abdominal pain, distension, and jaundice for 4 weeks. Imaging showed a diffuse process in the liver. Liver biopsy revealed adenocarcinoma infiltration of the liver.
2. A 58-year-old female with diabetes and elevated liver enzymes was evaluated. She had a history of elevated enzymes attributed to lipid medication years ago. Current labs showed elevated AST and ALT with normal ALP and GGT. She had weight gain and abnormal lipid profile.
3. The first case describes a patient with diffuse liver lesions found to be metastatic adenocarcinoma on biopsy. The second case involves a patient with metabolic risk factors and elevated amin
1) The study found that interactions between the nuclear receptor SHP and the transcription factor FOXA1 help maintain oscillatory homocysteine homeostasis in mice.
2) SHP was found to inhibit the transcriptional activation of genes involved in homocysteine metabolism (Bhmt and Cth) by FOXA1.
3) Mice lacking SHP had altered timing in the expression of homocysteine metabolism genes and differences in metabolite levels related to homocysteine metabolism compared to normal mice.
Metabolic changes in diabetes result from insulin deficiency and profoundly affect the liver, muscle, and adipose tissue. This leads to hyperglycemia and ketoacidosis as the liver produces glucose while tissues cannot utilize it. It also causes increased mobilization of fatty acids from adipose tissue and their breakdown in the liver, resulting in the production of ketone bodies. Excess fatty acids are also converted to triglycerides and secreted into the bloodstream. The insulin deficiency shifts energy production away from carbohydrate metabolism and towards fat oxidation, cholesterol synthesis, and ketone body synthesis.
Alcoholic hepatitis is a common condition caused by heavy alcohol consumption that carries a high mortality risk. Key aspects include:
- Presentation includes jaundice, fever, tender hepatomegaly and abnormal liver function tests.
- Severity is assessed using Maddrey's discriminant function, with scores over 32 indicating poor prognosis.
- Treatment of severe cases involves corticosteroids to reduce immune-mediated injury, pentoxifylline to inhibit tumor necrosis factor production, and nutritional support to address negative nitrogen balance and increased energy needs.
- Corticosteroids and pentoxifylline have been shown to improve short-term survival in randomized controlled trials for patients with severe disease.
Unlocking Diabetic Nephropathy (DN) through its key pathological mechanisms - Oxidative Stress and Fibrosis
https://coboscientific.com/biomarkers/diabetic-nephropathy/
The document discusses non-alcoholic fatty liver disease (NAFLD) and its relationship to metabolic syndrome. It begins by defining NAFLD and its subtypes, including simple steatosis and non-alcoholic steatohepatitis (NASH). It then discusses the risk factors and pathophysiology of NAFLD, noting its association with obesity, diabetes, and other components of metabolic syndrome. The document outlines current diagnostic and treatment approaches for NAFLD, including lifestyle modifications involving diet, exercise and weight loss. It also discusses potential drug therapies and newer treatment strategies being explored.
This document classifies and describes different types of inherited and acquired lipoprotein disorders. It discusses primary hyperlipoproteinemias caused by genetic defects affecting lipoprotein metabolism and transport. Secondary lipoprotein disorders are acquired conditions caused by diseases like diabetes, nephrotic syndrome, and atherosclerosis. The document outlines different types of hyperlipidemias classified based on elevated lipid fractions, including hypercholesterolemia, hypertriglyceridemia, and combined hyperlipidemia. Complications like xanthomas and atherosclerosis are discussed. Specific genetic disorders affecting apolipoproteins and resulting in abnormal lipid profiles are also described.
This document provides an overview of lipid metabolism and hypolipidemic drugs. It discusses lipids, lipoproteins, lipoprotein classification, lipid metabolism pathways including exogenous and endogenous pathways, atherogenesis, and drug therapy. The endogenous pathway can be separated into the atherogenic apo-B100 lipoprotein system and the antiatherogenic apo-A1 lipoprotein system. The document provides details on the structure and function of lipoproteins, their classification based on density and electrophoretic mobility, and the roles of apolipoproteins.
Metabolic syndrome is a collection of risk factors that increases the risk of cardiovascular disease and diabetes. It involves having at least three of the following: central obesity, high blood pressure, high blood sugar, high triglycerides, or low HDL cholesterol. An estimated 50 million Americans have metabolic syndrome. Risk factors include obesity, physical inactivity, stress, aging, and genetic predisposition. Symptoms include central obesity, high blood pressure, and abnormal cholesterol and blood sugar levels. Management focuses on lifestyle changes like diet, exercise, weight loss, and medication to control risk factors. Complications can include heart disease, stroke, diabetes, and kidney disease.
This study investigated the role of gut microbiota and bile acid metabolism in the development of non-alcoholic steatohepatitis (NASH)-associated hepatocellular carcinoma (HCC) in mice fed a high-fat diet. The mice developed NASH and HCC within 9 weeks of being fed the steatohepatitis-inducing diet without carcinogens. The gut microbiota was found to promote secondary bile acid production, which activated mTOR signaling in hepatocytes and contributed to oncogenesis. The findings suggest that metabolites produced by gut microbiota like secondary bile acids may mediate HCC development in NASH through mTOR pathway activation in liver cells.
Hyperlipidemia refers to elevated levels of lipids in the blood, including triglycerides and cholesterol. There are several types of hyperlipidemia based on the elevated lipoprotein, including types I-V. Type I is caused by deficiencies of lipoprotein lipase or ApoC-II, resulting in an inability to remove chylomicrons from the blood. Type IIa, or familial hypercholesterolemia, is caused by mutations in the LDL receptor gene and results in markedly elevated LDL cholesterol. It can be either homozygous or heterozygous. Both forms increase the risk of premature cardiovascular disease if not properly treated.
Use of Tumor Markers in Liver, Bladder, Cervical, and Gastric CancersLAB IDEA
This chapter discusses tumor markers for liver cancer. Liver cancer, or hepatocellular carcinoma, is a major cause of cancer death worldwide. Early detection is important for effective treatment but many cases are asymptomatic and detected late. The guidelines evaluate tumor markers like alpha-fetoprotein for surveillance of high-risk patients and diagnosis of liver cancer, noting their limitations. Recommendations are provided on the appropriate use of tumor markers in conjunction with imaging for managing liver cancer.
The document discusses aminotransferases, which are enzymes found mainly in the liver that help catalyze reactions between amino acids and alpha-keto acids. Aminotransferases are important for synthesizing amino acids and proteins. They are also clinically significant as indicators of liver damage. Specifically, elevated levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the blood can signify liver injury. The document outlines normal AST and ALT levels and various diseases and conditions that can cause their elevation, including viral hepatitis, alcohol abuse, and certain medications. It recommends further evaluation and treatment of any underlying cause if AST and ALT levels are found to be high.
1. Steatosis, or fatty liver, is common and usually benign but can progress to steatohepatitis if associated with conditions like obesity, diabetes, or alcohol use.
2. Alcohol is a leading cause of steatohepatitis and the most common type of liver disease in Western nations, due to the toxic byproducts produced when the liver metabolizes alcohol.
3. The pathology of alcohol-related liver disease ranges from reversible fatty liver to fibrosis, cirrhosis, and even liver cancer. Management involves cessation of alcohol and treatment of complications.
This document provides an overview of metabolic pathways and inborn errors of metabolism. It discusses the major metabolic pathways for carbohydrates, proteins, and lipids and how genetic defects can disrupt these pathways. It also describes the three main types of inborn errors of metabolism and provides examples. The document then goes into more detail about carbohydrate metabolism, including glycolysis, pyruvate metabolism, and the tricarboxylic acid cycle. It also discusses glycogen metabolism, protein metabolism, fat metabolism, and approaches to investigating metabolic cases.
This document provides an overview of diabetes mellitus, including its classification, pathophysiology, clinical features, investigations, diagnostic criteria, and management. It discusses the different types of diabetes, risk factors, characteristics, and laboratory findings. Type 1 diabetes results from beta cell destruction leading to insulin deficiency, while type 2 involves insulin resistance with relative insulin deficiency. Gestational diabetes occurs during pregnancy.
This document reviews the cellular mechanisms and clinical evidence regarding curcumin's effects on oxidative stress-related liver diseases. It finds that curcumin exerts remarkable protective and therapeutic effects through various mechanisms, including suppressing proinflammatory cytokines and lipid peroxidation, activating protective signaling pathways, and ameliorating cellular responses to oxidative stress like increasing antioxidant enzyme expression. Curcumin itself acts as a free radical scavenger against various reactive oxygen species due to its phenolic, β-diketone, and methoxy functional groups. Further clinical studies are still needed to better understand curcumin's structure-activity relationships and mechanisms in oxidative liver diseases.
Diabetes mellitus, its types and compicationsMohit Adhikary
This document discusses types and complications of diabetes mellitus. It begins with an outline that defines diabetes and classifies diabetes types and complications as acute or chronic. It then discusses the various types of diabetes in more detail, including type 1 diabetes pathogenesis and genetic and environmental risk factors. Type 2 diabetes risk factors and pathophysiology involving insulin resistance and secretion are covered. Other specific rare genetic types are defined. The document concludes by examining acute complications like diabetic ketoacidosis and chronic complications involving microvascular and macrovascular involvement, as well as theories around how hyperglycemia may lead to these complications. Glycemic control studies proving the benefits of control are also summarized.
Diabetes mellitus (DM) refers to a group of common metabolic disorders that share the phenotype of hyperglycemia, due to defect in insulin secretion, insulin action or both .
Beta oxidation defects are inborn errors of metabolism that result in the failure of fatty acid oxidation. There are several types depending on the length of the fatty acid involved: very long chain, long chain, medium chain, or short chain. The main pathways affected are carnitine transport, beta oxidation, and ketogenesis. Presentations range from hypoglycemia in infants to muscle weakness or exercise intolerance in older children and adults. Diagnosis involves testing for intermediary metabolites, acylcarnitines, and enzyme activities. Treatment focuses on avoiding fasting and providing rapid glucose treatment during acute illnesses.
This document discusses diabetes mellitus (DM), including:
1) DM is a chronic disease characterized by high blood sugar levels due to either the pancreas not producing enough insulin or the body not properly using the insulin produced.
2) There are two main types of DM - type 1 is usually diagnosed in childhood/early adulthood and results from autoimmune destruction of insulin-producing beta cells, while type 2 accounts for 80% of cases and involves insulin resistance and relative lack of insulin.
3) Uncontrolled DM can lead to serious damage of various organs and body systems, and was responsible for an estimated 1.5 million deaths in 2012. Lifestyle changes such as healthy diet, exercise and weight control can help
The document discusses calcium, phosphate, and magnesium content and distribution in the human body. It also describes the physiological functions of calcium, including cellular functions like cell growth and division, and extracellular functions like mineralization and blood clotting. Calcium signaling is discussed, where calcium binds to and activates proteins like calmodulin and calcium ATPase pumps.
1. A 65-year-old male presented with fever, abdominal pain, distension, and jaundice for 4 weeks. Imaging showed a diffuse process in the liver. Liver biopsy revealed adenocarcinoma infiltration of the liver.
2. A 58-year-old female with diabetes and elevated liver enzymes was evaluated. She had a history of elevated enzymes attributed to lipid medication years ago. Current labs showed elevated AST and ALT with normal ALP and GGT. She had weight gain and abnormal lipid profile.
3. The first case describes a patient with diffuse liver lesions found to be metastatic adenocarcinoma on biopsy. The second case involves a patient with metabolic risk factors and elevated amin
1) The study found that interactions between the nuclear receptor SHP and the transcription factor FOXA1 help maintain oscillatory homocysteine homeostasis in mice.
2) SHP was found to inhibit the transcriptional activation of genes involved in homocysteine metabolism (Bhmt and Cth) by FOXA1.
3) Mice lacking SHP had altered timing in the expression of homocysteine metabolism genes and differences in metabolite levels related to homocysteine metabolism compared to normal mice.
Metabolic changes in diabetes result from insulin deficiency and profoundly affect the liver, muscle, and adipose tissue. This leads to hyperglycemia and ketoacidosis as the liver produces glucose while tissues cannot utilize it. It also causes increased mobilization of fatty acids from adipose tissue and their breakdown in the liver, resulting in the production of ketone bodies. Excess fatty acids are also converted to triglycerides and secreted into the bloodstream. The insulin deficiency shifts energy production away from carbohydrate metabolism and towards fat oxidation, cholesterol synthesis, and ketone body synthesis.
Alcoholic hepatitis is a common condition caused by heavy alcohol consumption that carries a high mortality risk. Key aspects include:
- Presentation includes jaundice, fever, tender hepatomegaly and abnormal liver function tests.
- Severity is assessed using Maddrey's discriminant function, with scores over 32 indicating poor prognosis.
- Treatment of severe cases involves corticosteroids to reduce immune-mediated injury, pentoxifylline to inhibit tumor necrosis factor production, and nutritional support to address negative nitrogen balance and increased energy needs.
- Corticosteroids and pentoxifylline have been shown to improve short-term survival in randomized controlled trials for patients with severe disease.
Unlocking Diabetic Nephropathy (DN) through its key pathological mechanisms - Oxidative Stress and Fibrosis
https://coboscientific.com/biomarkers/diabetic-nephropathy/
The document discusses non-alcoholic fatty liver disease (NAFLD) and its relationship to metabolic syndrome. It begins by defining NAFLD and its subtypes, including simple steatosis and non-alcoholic steatohepatitis (NASH). It then discusses the risk factors and pathophysiology of NAFLD, noting its association with obesity, diabetes, and other components of metabolic syndrome. The document outlines current diagnostic and treatment approaches for NAFLD, including lifestyle modifications involving diet, exercise and weight loss. It also discusses potential drug therapies and newer treatment strategies being explored.
This document classifies and describes different types of inherited and acquired lipoprotein disorders. It discusses primary hyperlipoproteinemias caused by genetic defects affecting lipoprotein metabolism and transport. Secondary lipoprotein disorders are acquired conditions caused by diseases like diabetes, nephrotic syndrome, and atherosclerosis. The document outlines different types of hyperlipidemias classified based on elevated lipid fractions, including hypercholesterolemia, hypertriglyceridemia, and combined hyperlipidemia. Complications like xanthomas and atherosclerosis are discussed. Specific genetic disorders affecting apolipoproteins and resulting in abnormal lipid profiles are also described.
This document provides an overview of lipid metabolism and hypolipidemic drugs. It discusses lipids, lipoproteins, lipoprotein classification, lipid metabolism pathways including exogenous and endogenous pathways, atherogenesis, and drug therapy. The endogenous pathway can be separated into the atherogenic apo-B100 lipoprotein system and the antiatherogenic apo-A1 lipoprotein system. The document provides details on the structure and function of lipoproteins, their classification based on density and electrophoretic mobility, and the roles of apolipoproteins.
Metabolic syndrome is a collection of risk factors that increases the risk of cardiovascular disease and diabetes. It involves having at least three of the following: central obesity, high blood pressure, high blood sugar, high triglycerides, or low HDL cholesterol. An estimated 50 million Americans have metabolic syndrome. Risk factors include obesity, physical inactivity, stress, aging, and genetic predisposition. Symptoms include central obesity, high blood pressure, and abnormal cholesterol and blood sugar levels. Management focuses on lifestyle changes like diet, exercise, weight loss, and medication to control risk factors. Complications can include heart disease, stroke, diabetes, and kidney disease.
This study investigated the role of gut microbiota and bile acid metabolism in the development of non-alcoholic steatohepatitis (NASH)-associated hepatocellular carcinoma (HCC) in mice fed a high-fat diet. The mice developed NASH and HCC within 9 weeks of being fed the steatohepatitis-inducing diet without carcinogens. The gut microbiota was found to promote secondary bile acid production, which activated mTOR signaling in hepatocytes and contributed to oncogenesis. The findings suggest that metabolites produced by gut microbiota like secondary bile acids may mediate HCC development in NASH through mTOR pathway activation in liver cells.
Hyperlipidemia refers to elevated levels of lipids in the blood, including triglycerides and cholesterol. There are several types of hyperlipidemia based on the elevated lipoprotein, including types I-V. Type I is caused by deficiencies of lipoprotein lipase or ApoC-II, resulting in an inability to remove chylomicrons from the blood. Type IIa, or familial hypercholesterolemia, is caused by mutations in the LDL receptor gene and results in markedly elevated LDL cholesterol. It can be either homozygous or heterozygous. Both forms increase the risk of premature cardiovascular disease if not properly treated.
Use of Tumor Markers in Liver, Bladder, Cervical, and Gastric CancersLAB IDEA
This chapter discusses tumor markers for liver cancer. Liver cancer, or hepatocellular carcinoma, is a major cause of cancer death worldwide. Early detection is important for effective treatment but many cases are asymptomatic and detected late. The guidelines evaluate tumor markers like alpha-fetoprotein for surveillance of high-risk patients and diagnosis of liver cancer, noting their limitations. Recommendations are provided on the appropriate use of tumor markers in conjunction with imaging for managing liver cancer.
The document discusses aminotransferases, which are enzymes found mainly in the liver that help catalyze reactions between amino acids and alpha-keto acids. Aminotransferases are important for synthesizing amino acids and proteins. They are also clinically significant as indicators of liver damage. Specifically, elevated levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the blood can signify liver injury. The document outlines normal AST and ALT levels and various diseases and conditions that can cause their elevation, including viral hepatitis, alcohol abuse, and certain medications. It recommends further evaluation and treatment of any underlying cause if AST and ALT levels are found to be high.
Biochemical markers in diagnosis of Liver DIseaseChee Oh
This document discusses liver enzyme tests, what they measure, normal ranges, and their clinical significance in evaluating liver health and disease. It focuses on aminotransferases ALT and AST, which are released when liver cells are damaged and are markers of hepatocellular injury. Elevations in ALT are more specific to the liver, while AST is also found in other tissues. The ratio of AST to ALT and the magnitude of elevation provides clues to different liver diseases.
This document summarizes key details about aminotransferases and alkaline phosphatase enzymes. It discusses how aminotransferases like aspartate aminotransferase (AST) and alanine aminotransferase (ALT) catalyze the transfer of amino groups between amino acids and ketoacids, and how their levels are measured to monitor liver damage. AST is more elevated in conditions like hepatitis while ALT is higher in viral hepatitis. Alkaline phosphatase requires magnesium as an activator and exists as isoenzymes in the liver, bone, placenta and intestine that can be differentiated through electrophoresis or immunochemical methods.
Liver cancer diagnostics and Future trendsThet Su Wynn
Liver cancer is usually diagnosed through a combination of tumor marker tests, imaging scans, and biopsy. Serum markers like AFP, AFP-L3, and PIVKA-II provide early screening but lack specificity. Ultrasound, CT, and MRI scans use contrast agents to detect hypervascular lesions during arterial wash-in and wash-out, but are not definitive. Biopsy confirms the diagnosis by examining tissue samples for liver cell abnormalities, fibrosis, and marker expression through histology and IHC staining. Future areas of research include improved non-invasive diagnostic tools using genetic markers like miRNA profiles and tumor-specific genes.
The document discusses liver function tests (LFTs) and their use in evaluating liver diseases. It provides details on 3 key LFTs:
1. Bilirubin tests which are used to diagnose prehepatic (hemolytic), hepatic, and obstructive jaundice. Elevated conjugated bilirubin indicates obstructive jaundice while elevated unconjugated bilirubin indicates hepatic or hemolytic jaundice.
2. Liver enzymes like ALT, AST, ALP, and GGT which provide information on liver health and injury. Elevated ALT and AST indicate liver parenchymal damage while elevated ALP and GGT can indicate obstructive jaundice.
3
Nonalcoholic fatty liver disease (NAFLD) refers to hepatic steatosis without other causes like heavy alcohol use. It ranges from nonalcoholic fatty liver (NAFL) to nonalcoholic steatohepatitis (NASH) with inflammation. NAFLD prevalence is increasing worldwide in line with obesity, diabetes, and metabolic syndrome. Weight loss through lifestyle changes and possibly bariatric surgery is the only treatment with evidence of benefit for NAFLD/NASH.
Hígado graso no alcohólico en niños y adolescentes obesosCuerpomedicoinsn
The document discusses non-alcoholic fatty liver disease (NAFLD) and its relationship to obesity and insulin resistance. It notes that NAFLD affects around 20% of adults and 5% of children, and is strongly associated with obesity. A small portion of NAFLD cases progress to non-alcoholic steatohepatitis (NASH), which can potentially lead to scarring, cirrhosis, and liver failure over many years if not treated or reversed. Lifestyle changes such as weight loss and increased physical activity are recommended for treatment.
This document discusses nonalcoholic fatty liver disease (NAFLD). It begins by explaining that NAFLD ranges from simple steatosis to nonalcoholic steatohepatitis (NASH), which can progress to cirrhosis. Insulin resistance plays a key role in the metabolic abnormalities seen in NAFLD. The pathogenesis of NASH is not fully understood. Currently, there are no approved therapies, so treatment focuses on lifestyle modifications like weight loss and exercise to improve comorbidities. The prevalence of NAFLD is increasing due to the rising obesity epidemic.
This document discusses nonalcoholic fatty liver disease (NAFLD), its causes and risk factors. NAFLD is caused by fat accumulation in the liver from factors other than alcohol consumption, including genetic predisposition, environmental stresses, and metabolic stresses. The main causes discussed are insulin resistance, lipid metabolism disorders, mitochondrial dysfunction, and oxidative stress. Prevention and treatment involves lifestyle changes like a healthy diet and exercise, as well as medication to treat underlying conditions and reduce liver damage.
NAFLD non alcoholic fatty liver disease.pptxSyedFurqan30
This document provides an overview of non-alcoholic fatty liver disease (NAFLD). It discusses that NAFLD is increasingly a cause of liver disease globally, with a prevalence of 25% worldwide and 9-53% in India. NAFLD refers to hepatic fat accumulation in the absence of excessive alcohol use or other known liver diseases. The spectrum of NAFLD ranges from simple steatosis to non-alcoholic steatohepatitis (NASH) which is characterized by inflammation and cellular injury, and can progress to cirrhosis. Genetic and environmental factors contribute to NAFLD pathogenesis. Insulin resistance is a major driver, and comorbidities like obesity, diabetes, and obstructive
Non Alcoholic Fatty Liver Disease: A New Urban Epidemic.KETAN VAGHOLKAR
This document discusses non-alcoholic fatty liver disease (NAFLD), which has become very common in urban populations. NAFLD ranges from simple fatty liver to non-alcoholic steatohepatitis (NASH), which is characterized by fatty changes, inflammation, and fibrosis that can progress to cirrhosis. The main causes are obesity, insulin resistance, and dyslipidemia. Weight loss and improving insulin sensitivity through diet and exercise are the primary treatment approaches. Medications like vitamin E, pioglitazone, and metformin may also provide benefits, but more research is still needed on medical therapies for NAFLD.
The document discusses non-alcoholic fatty liver disease (NAFLD), which includes a spectrum of conditions from simple steatosis to non-alcoholic steatohepatitis (NASH) and cirrhosis. NAFLD is strongly associated with obesity and metabolic syndrome. The prevalence of NAFLD is increasing globally and varies from 5-30% in different regions. Diagnosis requires imaging and liver biopsy. Treatment focuses on lifestyle modifications and medications to improve insulin resistance.
The document discusses metabolic syndrome (MetS), also known as syndrome X. MetS is defined as a constellation of interconnected factors that increase the risk of cardiovascular disease, diabetes, and mortality. It commonly includes central obesity, elevated blood pressure, high blood glucose, excess body fat around the waist. The pathophysiology involves chronic low-grade inflammation and insulin resistance linked to obesity, genetic susceptibility, and lifestyle factors. Treatment focuses on lifestyle modifications like weight loss, dietary changes, and physical activity as well as medications.
Non-alcoholic fatty liver disease (NAFLD) is the buildup of fat in the liver not caused by alcohol. It is the most common form of chronic liver disease, affecting 10-20% of children globally. Risk factors include obesity, diabetes, and metabolic syndrome. NAFLD can progress from a harmless fat buildup (steatosis) to a serious inflammation (NASH), and eventually fibrosis and cirrhosis if not addressed. The pathogenesis involves genetic predisposition combined with environmental factors like diet, gut bacteria, and insulin resistance that together cause liver damage over time.
1. Alcoholic liver disease progresses through stages including fatty liver, alcoholic hepatitis, and cirrhosis. Tumor necrosis factor (TNF) plays a key role in mediating inflammation and the progression of disease.
2. Only a minority of heavy drinkers develop serious liver damage, and other host and environmental factors determine progression. Risk factors include genetic factors and additional insults like hepatitis or obesity.
3. TNF is involved in the early stages of fatty liver disease and mediates the transition to more advanced stages like steatohepatitis and cirrhosis. Cytokines like TNF orchestrate the liver's inflammatory response and damage.
Metabolic syndrome is one of the most common risk factor for Cardiovascular disease. Greek, unani, ayurvedic Herbal medicine shows great potential in helping fight the condition. in these presentation an attempt made to understand the pathophysiology in detail, and How Unani system of medicine address this whole syndrome along with the details of potent herbs which can be used for the Metabolic syndrome.
This document summarizes the effects of diet and exercise on liver lipid metabolism. It discusses the normal pathways of hepatic lipid metabolism, including lipid influx from adipose tissue and diet, de novo lipogenesis from carbohydrates, and lipid oxidation and secretion. Dietary fat and carbohydrates can accumulate in the liver as triglycerides if uptake and synthesis exceed oxidation and secretion. The roles of different transcription factors and enzymes involved in lipogenesis are also described.
Hepatotoxicity, or liver toxicity, can result from anti-tuberculosis (TB) drugs and is known as drug-induced hepatitis (DIH). Patients at high risk include those with pre-existing liver conditions, alcohol use, and advanced TB. Monitoring of liver enzymes is important for high risk patients during TB treatment. Symptoms of DIH include fatigue, nausea, and jaundice. Diagnosis involves abnormal liver enzymes and symptom resolution after stopping anti-TB drugs. Management consists of gradual dose escalation while monitoring for toxicity.
Total Bile Acids - The Importance of Fifth Generation TestsRandox Reagents
Measuring total bile acid (TBA) levels may prove useful for the detection of liver diseases such as viral hepatitis, mild liver injury through drug use and for further evaluation of patients with chronic hepatitis who were previously treated successfully. TBA levels may rise up to 100 times the normal concentration in patients with liver disease due to impairment of hepatic synthesis and extraction of bile acids.
Intrahepatic cholestasis of pregnancy (ICP) or obstetric cholestasis is a pregnancy-specific liver disorder. It can be indicated by pruritus, jaundice, elevated TBA levels and/or serum transaminases and usually affects women during the second and third trimester of pregnancy.
Newer methods such as the enzyme cycling method or fifth generation methods offer many advantages including greater sensitivity, liquid reagents, small sample volumes and reduced instrument contamination from formazan dye. Additionally, the fifth generation assay does not suffer from interference from lipaemic or haemolytic samples. Both lipaemia and haemolysis are common in new-borns and pregnant women.
This study assessed clinical, virological, histopathological and biochemical factors related to hepatic steatosis in Egyptian patients with HCV genotype 4. The study found steatosis in 80% of early HCV patients and 88% of cirrhotic patients. Steatosis grade correlated with higher viral load and liver inflammation. Patients had lower triglycerides and cholesterol levels correlated with steatosis grade. They also had higher leptin and beta-2-microglobulin levels and lower adiponectin levels compared to controls, correlating with steatosis grade and liver disease severity. The results suggest HCV genotype 4 is associated with biochemical changes like increased leptin and beta-2-microglobulin that may
Acute fatty liver of pregnancy (AFLP) is a rare but potentially fatal liver condition that can occur late in pregnancy. It is caused by an abnormal breakdown of fatty acids in the liver due to genetic mutations. AFLP presents with nausea, abdominal pain, and liver dysfunction. The condition is diagnosed based on clinical criteria and liver biopsy, and is treated by delivering the baby to reverse liver damage, followed by supportive care which may include intensive care, coagulation treatment, and glucose management. While still risky, early diagnosis and treatment have improved outcomes for both mother and baby.
Pósters presentados al congreso Cientifícatebardasco
Póster que hemos presentado al congreso cientifícate del años 2019. Han participado nuestros alumnos de 2º, 3º y 4º de ESO con cinco proyectos de investigación.
Este documento presenta los resultados de un estudio realizado por estudiantes de 4o de ESO para determinar la cantidad de bacterias presentes en diferentes lugares del Colegio Ramón y Cajal. Los estudiantes muestrearon 29 lugares y cultivaron las muestras para contar las colonias bacterianas. Los lugares con mayor número de bacterias fueron el suelo del patio y el césped artificial, probablemente debido a su ubicación al aire libre y al tránsito de personas.
Este protocolo describe los métodos para identificar muestras de ADN mediante el análisis de dos loci polimórficos: la secuencia Alu y el locus VNTR D1S80. Se extrae ADN de muestras bucales y se amplifica cada locus mediante PCR. Los productos de PCR se analizan mediante electroforesis en gel de agarosa para determinar los genotipos de cada individuo y compararlos.
Este documento describe las propiedades fundamentales de la materia y las magnitudes físicas. Explica que la materia tiene masa y volumen, y que las sustancias se distinguen por sus propiedades específicas. También define las magnitudes fundamentales como longitud, masa y tiempo, y las magnitudes derivadas como velocidad, superficie y densidad. Finalmente, detalla cómo medir estas propiedades usando el Sistema Internacional de Unidades.
Este documento describe los reinos Protoctistas, Moneras y los virus. El reino Protoctistas incluye protozoos unicelulares y algas. Los protozoos son microscópicos y heterótrofos, mientras que las algas son autótrofas. El reino Moneras incluye bacterias unicelulares procariotas. Los virus no son considerados seres vivos y solo pueden reproducirse infectando células.
Este documento describe las características generales de las plantas y los hongos. Explica que las plantas están compuestas de células con paredes de celulosa, cloroplastos y realizan la fotosíntesis. Se clasifican en plantas sin flores como musgos, hepáticas y helechos, y plantas con flores como gimnospermas y angiospermas. También describe los procesos de nutrición, reproducción y estructura básica de las plantas. En cuanto a los hongos, señala que viven en medi
Este documento describe las características de varios grupos de animales. Los animales son eucariotas pluricelulares que se alimentan de otros organismos (heterótrofos) y tienen órganos sensoriales y capacidad de movimiento. Se dividen en vertebrados e invertebrados. Entre los invertebrados se encuentran poríferos, celentéreos, platelmintos, nematodos, anélidos, moluscos, artrópodos y equinodermos.
El documento describe las características de los diferentes grupos de vertebrados. Explica que los vertebrados se dividen en animales vertebrados e invertebrados, y que los vertebrados se caracterizan por tener una columna vertebral y se clasifican en mamíferos, aves, reptiles, anfibios y peces. También describe algunas de las características distintivas de cada grupo.
Los seres vivos comparten características como estar constituidos por células, realizar funciones vitales como la nutrición, relación y reproducción, y estar formados por sustancias químicas como el carbono, hidrógeno, oxígeno y nitrógeno. Todos los seres vivos, desde animales, plantas, hongos y microorganismos están compuestos de células, que son la unidad básica de la vida.
Las rocas están formadas por minerales. Los minerales pueden apreciarse a simple vista en algunas rocas, mientras que en otras no se pueden apreciar. La resistencia de una roca depende del mineral del que esté formada; los minerales resistentes forman rocas resistentes y los minerales frágiles forman rocas frágiles.
Este documento describe las propiedades y clasificación de los minerales. Explica que los minerales son sustancias naturales con composición química específica y que forman parte de las rocas. Los minerales más comunes contienen oxígeno y silicio, llamados silicatos, pero también existen minerales sin silicio. Detalla diferentes tipos de silicatos y no silicatos, y cómo se originan los minerales a través de procesos geológicos.
El documento describe la distribución y propiedades del agua en la Tierra. La mayor parte del agua es salada en los océanos, mientras que una pequeña parte es agua dulce en ríos, lagos y glaciares. El agua se mueve continuamente a través del ciclo del agua entre la atmósfera y la superficie a través de la evaporación y la precipitación.
it describes the bony anatomy including the femoral head , acetabulum, labrum . also discusses the capsule , ligaments . muscle that act on the hip joint and the range of motion are outlined. factors affecting hip joint stability and weight transmission through the joint are summarized.
Thinking of getting a dog? Be aware that breeds like Pit Bulls, Rottweilers, and German Shepherds can be loyal and dangerous. Proper training and socialization are crucial to preventing aggressive behaviors. Ensure safety by understanding their needs and always supervising interactions. Stay safe, and enjoy your furry friends!
A review of the growth of the Israel Genealogy Research Association Database Collection for the last 12 months. Our collection is now passed the 3 million mark and still growing. See which archives have contributed the most. See the different types of records we have, and which years have had records added. You can also see what we have for the future.
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
This presentation includes basic of PCOS their pathology and treatment and also Ayurveda correlation of PCOS and Ayurvedic line of treatment mentioned in classics.
1. ASSOCIATION STUDY AND
FUNCTIONAL VALIDATION OF
GENETIC MARKERS RELATED TO
NON-ALCOHOLIC FATTY LIVER
DISEASES
Angel Bardasco Blazquez
(Tutor: Ana M. Aransay, CIC bioGUNE)
0
2. ABSTRACT
Non-alcoholic fatty liver diseases (NAFLD) are the most common causes of
chronic liver disease in several western countries. Up to now, the only way to diagnose
NAFLD with certainty is liver biopsy. Therefore, the aim of the present project is to find
genetic markers that could help to develop a non-invasive diagnostic method for these
disorders. To achieve this objective we have carried out a candidate gen association
approach that yielded 6 potentially associated genes, being the first time that SLC2A1
has been associated with NAFLD. In addition, we have investigated the regulation of
those genes in liver biopsies at transcriptomic level. We have detected that all the
studied genes are regulated in NAFLD, and that the regulation is higher in advanced
stages of the disease. This suggests that the identified genes could serve as potential
markers for the diagnosis of the phases of NAFLD development.
1
3. INTRODUCTION
Non-alcoholic fatty liver disease (NAFLD) includes a wide spectrum of lesions
including steatosis (ST), non-alcoholic steatohepatitis (NASH), fatty liver and
inflammation, as well as a high number of cryptogenic cirrhoses 1. NAFLD represents
the hepatic manifestation of the metabolic syndrome with insulin resistance as a
common feature, namely, central obesity, insulin resistance, dyslipidemia and
hypertension 2, 3. ST is an accumulation of fatty acids in the liver 4 and NASH is a term
that describes a form of liver disease that is histologically indistinguishable from
4, 5
alcoholic hepatitis, but occurs in persons who do not consume excess of ethanol
(less than 40g of ethanol per week 3).
Prevalence of NAFLD
NAFLD is the most common cause of chronic liver disease in several western
countries 5 and it has been reported in all age groups including children, although the
highest prevalence is described in individuals between 40 and 60 years 3. About 75% of
obese patients have NAFLD 6 (the prevalence augments with increasing body weight 3).
In general population, the prevalence of NAFLD is estimated about 20% 7. The
prevalence of NAFLD is increasing in industrialized countries 2 due probably to social
and environmental agents like alcohol, industrial toxins and hepatotrophic viruses 5,
and also to metabolic syndromes like obesity and type 2 diabetes (T2D) 8. Nugent et al.
6
showed that patients with T2D have high risk to develop NAFLD, since NAFLD was
detected in 62% of patients with newly diagnosed T2D and T2D has been described in
34% to 75% of patients with NASH 3. Obesity and diabetes mellitus are not
predisposing factors only to develop NAFLD, but they are also potential risk factors to
develop severe hepatic fibrosis and cirrhosis 9. All of these previous mentioned factors,
hyperlidipemia and other conditions associated with insulin resistance are generally
present in patients with NAFLD 3: at the time of NASH diagnosis, up to one third of the
patients have diabetes or fasting hyperglycemia 4, between 39 and 100% are
overweighted or obese, and between 20 and 80% have abnormalities of lipid
metabolism 4.
2
4. Development and progression of NAFLD
The pathogenesis of NAFLD responds to a two-hit hypothesis. First of all, an
10
imbalance in fatty acid metabolism involves accumulation of fat in the liver
(steatosis), likely, as a result of insulin resistance and increased fat mass 5, 11. Secondly,
hepatocyte necrosis and apoptosis is driven by oxidative stress (and subsequent lipid
peroxidation), deregulated proinflammatory cytokines production by Kupffer cells
(principally the tumor necrosis factor alpha, TNFα) and hormones derived from
adipose tissue (adipocytokines) that result from efforts to compensate the altered lipid
homeostasis 1, 3, 11, 12 11. Jou et al.12 proposed that the hepatocyte death is the third and
decisive step in NAFLD pathogenesis because this event drives progression from NASH
to cirrhosis.
In the hepatic ST, vesicles of fat, predominately triglycerides, accumulate within
hepatocytes (without causing considerable hepatic inflammation) causing liver cell
5 12
death and activating mechanisms of hepatocyte regeneration . This regenerative
respond activates hepatic stellate cells to myofibroblasts, causing liver fibrosis and
expanding hepatic progenitor populations. Subsequently , several chemoattractans are
produced to recruit various types of immune cells into the liver, inducing hepatic
inflammation that drives to NASH 12. ST can progress to NASH due to hepatocyte injury
and apoptosis, and hepatic infiltration by inflammatory cells. It is unclear why some
patients who develop ST go on developing NASH while others do not 11. The next stage
could be that NASH develops to cirrhosis as a result of an incomplete repair of
metabolic liver injury 12. In most chronic liver diseases that lead to cirrhosis, there is an
increased risk of developing hepatocellular carcinoma (HCC) that is an irreversible
13
state of liver damage . Hepatocyte DNA damage and expansion of liver progenitor
cells have been demonstrated in early NASH and this suggests that NASH provides
fertile ground for neoplastic transformation of hepatocytes at several stages of
12
differentiation . Progression of simple ST to NASH increases the risk to develop
cirrhosis and consequent liver-related morbidity and mortality 12.
3
5. Figure 1. Lipid metabolism within the hepatocytes. Liver lipid content is determined
by the equilibrium of several processes: import of FFA from the adipose tissue, de
novo synthesis of FFA in hepatocytes, β-oxidation of FFAs, esterification of FFA into
triglycerides and export of triglycerides as vLDL. Hepatic ST is a consequence of
imbalance in those processes in favor of excessive triglyceride accumulation. Insulin
resistance and resulting hyperinsulinemia lead to hepatocyte lipid accumulation in
the liver by several mechanisms. In adipose tissue, insulin resistance enhances
triglyceride lipolysis and inhibits esterification of FFAs. The result is the increased
levels of circulating FFAs, which are then taken up by the liver. In hepatocytes, the
hyperinsulinemia increases the “de novo” synthesis of fatty acids and inhibits their β-
oxidation. The consequence of reduced vLDL production and triglyceride export is
the accumulation of FFAs within hepatocytes.
The normal lipid metabolism in the liver involves hepatocyte uptake and de
novo synthesis of free fatty acids (FFA), disposal of FFA via oxidation or de novo
triglyceride synthesis and export of triglycerides as a very low density lipoproteins
12
(vLDL) from hepatocytes (Figure 1) . As soon as the rate of triglyceride synthesis
overwhelms the capacity of vLDL export, triglycerides accumulate within hepatocytes
causing ST 12. When the insulin resistance occurs, hepatic FFA concentration increases
by the movement of FFA from adipocytes by lipolysis (and, consequently, increasing
hepatic import) and/or hepatic endogenous synthesis 4. Triglycerides by themselves
are not hepatotoxic but they are biomarkers of increased hepatic exposure to
potential toxic FFA. An enzyme for esterification (Acyl-coA:diacylglycerol
acylltransferase, DGAT) is required to transform FFA in triglycerides and to join vLDL12,
4
6. then, the capacity of vLDL export can be overwhelmed leading to FFA accumulation
within hepatocytes.
The molecular mechanism of the insulin resistance is complex and has not
been elucidated completely. Several molecules appear to interfere with the insulin
signaling pathway and it has been found that adiponectin plays a key role in insulin
3, 11
sensitivity . FFA and their metabolites are ligands of peroxisome proliferator-
activated receptor alpha (PPARα), a transcription factor that activates genes involved
in fatty acid oxidation. When PPARα is up-regulated, there is more FFA oxidation,
which is translated in increased oxidative stress, elimination of FFA and progression
from ST to NASH 3.
Oxidative stress
Hepatic mitochondria of patients with NASH exhibit ultrastructural lesions with
the presence of para-cristalline inclusions in the megamitochondria, while
14
mitochondria of patients with simple ST are normal . Although the mechanisms for
hepatic mitochondrial dysfunction in NASH are still unknown, it may involve lipid
peroxidation, TNFα and reactive oxygen species (ROS). TNFα increases permeability of
the mitochondrial membranes and the blocking of the electron flow from complex II to
14
complex IV . Those factors are supposed to alter mitochondrial DNA and
mitochondrial oxidative phosphorylation, producing the structural alterations
mentioned abobe3.
It has been shown that 30% of the patients with NASH have elevated ferritin
levels (marker of iron overload) 11, which plays a role in oxidative stress and may play a
15
function in pathogenesis of NASH . This iron overload generates reactive oxygen
species and subsequent lipid peroxidation. In addition, iron has harmful effects on the
mitochondria activity 11.
5
7. Diagnosis of NAFLD
Most of the diagnosis of NAFLD is done by exclusion of other liver diseases
taking into consideration parameters through a scoring system 11 like biochemical tests
based on serum markers, imaging techniques such as ultrasound, and measurement of
8, 13
liver stiffness by transient elastography . The gold standard for accurate diagnosis
of NAFLD is liver biopsy. This is the only way to distinguish between fatty liver
(steatosis) alone and NASH. There are not specific and sensitive noninvasive tests
3, 5
(there are some, but their efficiency has not been demonstrated) . The problems
with liver biopsy are that is painful, invasive and, given the increasing number of
patients with NAFLD, it is not an efficient method 8. Therefore, studies of these
diseases are limited by the inability to make a definitive diagnosis of NAFLD3.
Additionally, distinction of NASH from simple ST is important because their prognoses
and clinical management are different 8, 13. Studies that have used strict definitions for
diagnosis, including biopsies, were most often based on specific subsets of the
population (like diabetics, obese individuals, alcoholic liver diseases, etc.) and, so far,
they cannot be applied to the general population 3.
Treatment
Although there is no consensus treatment for ST and NASH, most of the applied
therapies include specific diet and exercise for weight loss and sometimes, it could be
16
enough to treat hepatic ST . It is also recommended to stop smoking and alcohol-
3
drinking during treatment . When specific diet and exercise is not enough,
pharmacological treatment should be initiated. Several drugs such as antioxidants and
3, 13
lipid-lowering drugs have been tried for the treatment of NAFLD . There is quite a
controversy about NAFLD pharmacological treatment because the response of patients
is variable and often there are important secondary effects.
6
8. Genetic of liver diseases
It has been shown that most evidences of genetic association with NAFLD
derive from family clustering analysis 2. There are different families of genes involved
in ST and NASH: genes influencing lipid metabolism, genes affecting oxidative stress,
genes coding for bacterial receptors and genes influencing extracellular matrix
synthesis and degradation 2. Modifications of those gDNA gene sequences have been
associated with liver diseases:
• The polymorphism (-493G/T) in the promotor of microsomal triglyceride
transfer protein (MTP) has been associated with NAFLD 17. This enzyme adds
triglycerides to nascent apolipoprotein B, producing vLDL. Thus, decreased
activity of MTP may lead to lipid accumulation 17.
• The (1183T/C) polymorphism in the manganese superoxide dismutase
(MnSOD), located in mitochondria and implicated in scavenging excessive
oxidative stress to hepatocytes, has been related with NAFLD 17.
• Polymorphisms (1031T/C) and (863C/A) in tumor necrosis factor alpha
(TNFα) were reported as associated with NASH in Japanese and Italian
population 18, 19. TNFα has been shown to induce insulin resistance, involved
in development of NASH.
• A SNP (V175M) in exon 8 of posphatidylethanolamine N-methyltransferase
(PEMT), that play a role in lipoprotein secretion from liver, has been seen
20
associated with NAFLD . This SNP it is a non-synonymous polymorphism
(aminoacid change: V175M) and generates a loss of function of PEMT.
• Mice exposed to a lipid rich diet developed severe NASH with fibrosis that
has been associated with overexpression of Cytochrome P450 2E1 (CYP2E1)
4
. Polymorphisms within this enzyme could be associated with risk of liver
disease.
• Mutations in gen hemochromatosis (HFE) (C282Y and H63D), related with
iron overload, have been associated with hepatic fibrosis 15.
7
9. • It was shown that polymorphism (667C/T) in the methylenetetrahydrofolate
reductase (MTHDR) has been associated with mayor risk to develop
21
hepatocellular carcinoma in patients with alcoholic cirrhosis , and some
alleles have been significantly associated with NASH 22.
23
Rubio et al. suggest that the alterations in gene expression associated with
NASH are broad and selective, and they found that many of the identified genes are
associated with mitochondrial function, insulin action and oxidative stress. Expression
of proliferator-activated receptor gamma (PPARγ) at mRNA level was significantly
24
lower in subjects with ST than in those without . Genes involved in scavenging of
reactive oxygen species (like catalase or glutathione peroxidase), as well as genes
involved in glucose (alcohol dehydrogenase 1 and glucose-6-phosphatase) and fatty
acid metabolism (like 3-hydroxy-3-,ethylglutaryl coenzyme A, mitochondrial 3-oxoacyl-
CoA thiolase and long-chain acyl-CoA synthetase) are down-regulated in NASH
25
patients . However, genes involved in protein synthesis, degradation pathways and
25
complement activation are up-regulated in NASH patients . It is interesting to
emphasize that patients with liver ST have a gene-expression pattern intermediate
23
between those patients with NASH and healthy controls . However, Rubio el al.23
described that all patients with ST do not develop NASH, and only those that have a
similar gene-expression pattern to the one associated to NASH seem to have a higher
risk to develop NASH.
Cytochrome P450 2E1 (CYP2E1) is up-regulated in patients with NASH while in
patients with ST is normal. Its activity has been associated with oxidative stress, insulin
12
resistance and hepatic lipid peroxidation . In addition, it has been seen that several
genes that are important for the mitochondrial function are down-regulated in NASH
patients 3.
8
10. Association Studies
The development of common diseases results from complex interactions
between numerous environmental factors and variation of several genes, and,
therefore, it is very interesting identifying the associated variations to understand the
biology of those diseases 26.
The Human Genome Project has deposited millions of Single Nucleotide
Polymorphisms (SNP) into public databases like dbSNP
(http://www.ncbi.nlm.nih.gov/sites/entrez) or International HapMap Project
27
(http://www.hapmap.org/) . The goal of the international HapMap Project is to
determine the common patterns of DNA sequence variation in the human genome and
to make this information freely available in the public domain. The data base contains
a map of these patterns across the genome by determining genotypes, their
28
frequencies and the degree of variability in different populations . The phase II
HapMap has characterized over 3.1 million of SNPs by genotyping 270 individuals from
four geographically diverse populations and includes 25-35% of common SNP variation
in the populations surveyed 29. In addition, HapMap Project characterized the linkage
disequilibrium (LD) patterns of different population based in the obtained genotypes.
LD means a nonrandom relationship of alleles at two or more loci that is inherited as
one single block. It is possible to do an association study of a significant proportion of
the common variation of a large number of genes that occurs in regions of high LD
where it is not necessary to genotype all SNPs within an LD-block but just one or two
26, 30
representatives of each region, which are called haplotype tag SNPs (htSNPs) .
When multiple markers in a chromosomal region are studied to assess the association
between this region and disease, a statistical analysis based on haplotypes may be
more informative than separate analyses of the individual markers 27.
Samples used for association should be selected with care:
• Case and control groups should be of the same ethnical population,
because if different population are mixed-up their different genetic
background can drive to false marker association (population
stratification) 26.
9
11. • The inclusion criteria for case and control selection should be very strict
according to their clinical parameters. The more phenotypical
information we have, the better.
The results obtained in a genetic association study should be validated by
testing the function of the associated genes within an in vivo system.
NAFLD association study at CIC bioGUNE
The present project is part of a study that is being carried out at CIC bioGUNE
for the association of genetic variations with NAFLD.
According to previous experimental studies (including knockout models,
transcriptomics, proteomics, and metabolomics), a list of candidate genes involved in
the pathogenesis of NASH was identified. Ninety two genes were considered according
to the following criteria: 62 genes were previously identified to be differentially
expressed in liver samples from patients with NASH and/or ST compared to controls 23;
17 genes are involved in hepatic One-Carbone metabolism, compromising the
methylation and folate cycles; and 13 genes had been associated to liver injury.
A total of 3,072 htSNPs were selected within those aforementioned candidate
genes based on the information available at international HapMap Project for the
European and Asian Populations.
Among all the samples obtained from the collaborative hospitals (Principe de
Asturias Hospital, Madrid, Spain; Clinic Hospital, Barcelona, Spain; Hospital de
Galdakao, Galdakao, Spain) only those with certain diagnosis for ST and/orNASH after
biopsy were genotyped. DNA from control individuals was purchased from the DNA
bank of BIOEF Foundation (Sondika, Spain). The inclusion criteria for the controls were
absence of Insulin Resistance Syndrome (no traces of hyperglycemia, hypertension or
obesity), normal liver activity tested by measuring the levels of transaminases and
Body Mass Index (BMI) ≤30 kg/m2.
10
12. A total of 69 patients and 217 controls were successfully genotyped by Golden
Gate technology following Illumina Inc.’s protocols.
Aim of the study
The general objective of CIC bioGUNE project is to find some genetic
association with non-alcoholic hepatic disease that can be used as a non-invasive
diagnosis method, following a candidate-gene association approach
The specific aim of the present work is to analyze the results of the association
study (statistics) and to test the mRNA expression level of the resulting associated
genes in liver biopsies of diagnosed individuals.
11
13. MATERIALS AND METHODS
Association study
Data obtained by GoldenGate Assay were decoded and corrected in Genome
Studio (2008 (c) Illumina, Inc. 2003-2008) software. Only good quality markers were
considered for further analysis.
Obtained genotypes and allele frequencies were compared between ST/NASH
cases and controls using PLINK Software v. 1.05. The analysis was done using allelic
test of single-marker and multi-marker association including all individuals. The data
filtering criteria were minor allele frequency (MAF) ≥ 0.01 and Hardy-Weinberg
equilibrium (HWE) ≥ 0.001.
Calculation of r2 and Gabriel et al. LD-block estimation31 were analyzed in
Haploview v. 4.1 (MAF ≥ 0.01 and HWE ≥ 0.001).
Differential expression
Total-RNA extracts were obtained from liver biopsies of control individuals, ST
and NASH patients. Human Universal Reference RNA (HUR) of Clontech (Stratagene:
740000) was used as a positive control.
Retro-transcription (RT) of samples and HUR was done following this protocol:
• 275 ng of total-RNA, 1µl of Oligo (dT) 12-18 (500µg/ml), 1µl dNTP mix
(10mM) and sterile distilled water were added per tube
• Mixture was heated at 65ºC for 5 min and contents were collected by brief
centrifugation
• 4µl of 5x First-Strand Buffer, 24µl of 0.1 M DTT and 1µl of RNaseOUTTM (40
units/µl) were added per tube
• Tubes were mixed gently and incubated at 42ºC for 2 min
12
14. • 1µl of SuperScriptTM (200 units, Invitrogen, Cat. No. 10777-019) was added
per tube and mixed
• Tubes were incubated at 42ºC for 50 min and 70ºC for 15min in order to
inactivate the enzyme
Then, 17µl of each cDNA product were diluted in 300µl of water.
cDNA of HUR was diluted as follows in order to have a standard curve of each
quantitative PCR (qPCR) reaction:
20µl of original cDNA + 400µl of distilled water
200µl of previous dilution + 200 µl of distilled water
200µl of previous dilution + 200 µl of distilled water
200µl of previous dilution + 200 µl of distilled water
200µl of previous dilution + 200 µl of distilled water
cDNA of cases and controls was analyzed in iCycler Thermal Cycler with iCycler
iQ Module developed by Bio-Rad.
Primers for measuring the mRNA expression of Cytochrome P450, family 2,
subfamily E, polypeptide 1 (CYP2E1), Serine/threonine kinase 11 (STK11), Solute carrier
family 2 (facilitated glucose transporter), member 1 (SLC2A1), Asparaginase synthetase
(ASNS) and 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR) were
purchased from Qiagen as QuantiTec® Primer Assay numbers QT01669962,
QT01008980, QT00068957, QT00084546, and QT00072156 respectively. The mRNA
expression of Glyceraldehyde-3-phosphate dehydrogenase (GADPH) and acidic
ribosomal phosphoprotein (ARP), as housekeeping genes, was analyzed using in-house
sets of primers.
For the qPCR, reactions were set in triplicates, and standard curve dilutions
were analyzed in duplicates. All reactions were done in a total volume of 20µl including
the following reagents:
13
15. For QuantiTec® Primer Assay primer sets (per tube):
1µl of EvaGreen 20x (Biotisem: 31000)
10µl of Hot Start MasterMix 2x (Metabion, mi-E8011)
2µl of Forward and Reverse primers 10X
5µl of cDNA sample
2µl of distilled water
For housekeeping genes primer sets (per tube):
1µl of EvaGreen 20x (Biotisem: 31000)
10µl of Hot Start MasterMix 2x (Metabion, mi-E8011)
0.54 µl of Forward primer 100mM
0.54 µl of Reverse primer 100mM
5µl of cDNA sample
2.92µl of distilled water
An automatic pipetting system (Eppendorff epMottion 5070) was used in order
to increase the reproducibility of the assays.The amplification cycling conditions for
the qPCR were:
• For ASNS, CYP2E1, STK11 and MTR: 40 cycles of 15s at 94ºC, 30s at 55ºC and
30s at 72ºC.
• For SLC2A1: 40 cycles of 15s at 94ºC, 30s at 61ºC and 30s at 72ºC.
• For housekeeping genes (GADPH and ARP): 40 cycles of 15s at 94ºC, 30s at 60ºC
and 30s at 72ºC.
Differential relative expression of each tested gene was estimated based on
the obtained Ct values, by the delta-delta-Ct method 32.
14
16. RESULTS
Association Study
A total of 1536 SNPs were successfully genotyped in all studied individuals (69
cases and 217 controls).
Eleven SNPs showed to be significantly associated (p < 10-4) with NAFLD for the
single-marker allelic test. These significant SNPs were located in the following genes:
Cytochrome P450, family 2, subfamily E, polypeptide 1 (CYP2E1), Smg-7 homolog,
nonsense mediated mRNA decay factor (SMG7), Solute carrier family 2 (facilitated
glucose transporter), member 1 (SLC2A1), 5-methyltetrahydrofolate-homocysteine
methyltransferase (MTR) and Serine/threonine kinase 11 (STK11) (see table 1). Seven
out of the eleven associated SNPs are located in SLC2A1 and all of them are in high LD
(figure 2).
CHR SNP gene A1 F_A F_U A2 CHISQ P Position
10 rs28969387 CYP2E1 A 0.063 0 T 27.52 1.56E-07 Exon 9
1 rs1044879 SMG7 G 0.627 0.406 C 17.43 2.98E-05 3'UTR
1 rs1770810 SLC2A1 A 0.278 0.127 G 16.4 5.13E-05 Intron
1 rs841856 SLC2A1 A 0.271 0.124 C 15.39 8.73E-05 Intron
1 rs3754255 MTR A 0.258 0.442 G 13.59 2.28E-04 Intron
19 rs7259033 STK11 G 0.33 0.521 C 12.93 3.24E-04 Intron
1 rs841858 SLC2A1 A 0.234 0.113 C 12.07 5.12E-04 Intron
1 rs4658 SLC2A1 G 0.286 0.152 C 11.71 6.23E-04 3'UTR
1 rs841848 SLC2A1 A 0.281 0.145 G 11.57 6.69E-04 Intron
1 rs3754223 SLC2A1 A 0.273 0.145 T 11.41 7.29E-04 Intron
1 rs2229682 SLC2A1 A 0.265 0.141 G 11 9.12E-04 Exon 6
-4
Table 1. Results of the single-marker analysis. Only significant (p < 10 ) SNPs are shown.
Multi-marker association was analyzed by sliding window from 2 to 10 SNPs-
windows, and p < 10-4 value was used as a threshold. This test revealed one group of 3
SNPs located in an intronic region of Asparaginase synthetase (ASNS) gene, which
resulted to be in total LD in the studied population (see table 2 and figure 2).
15
17. LOCUS gene start gene end HAPLOTYPE F_A F_U P Sliding windows
WIN1246 ASNS ASNS TAG 0.1176 0.2857 7.09E-05 rs7781469 rs4727377 rs7810919
-4
Table 2. Results of the sliding-windows analysis. Only significant (p < 10 ) windows are shown.
Figure 2. a) LD values (r2) of SLC2A1 (red color without number means 100% of LD) and b) of ASNS
significantly associated SNPs.
Differential expression
Differential expression analysis was done for all the genes represented by
several (more than one) associated SNPs with a p<10-2 (3 SNPs ofCYP2E1; 11 SNPs of
SLC2A1; 12 SNPs of MTR; 3 SNPs of STK11; 11 SNPs of ASNS. Therefore, although
rs1044879, which is a proxy of neutrophil cytosolic factor 2 (NCF2), was one of the
most significant SNPs (p=2.98 x 10-5), it was not considered for subsequent analysis
because NCF2 was represented uniquely by that polymorphism.
RT-qPCR for the 5 associated genes (CYP2E1, STK11, SLC2A1 ASNS MTR) and 2
housekeeping genes (GADPH and ARP) was carried out using the RNA extracted from
liver biopsies of a total of 5 ST and 5 NASH patients and 6 controls.
16
18. In figure 3 we represent the ratio (Cthousekeeping/Ct target gene) based on the media
Ct values obtained for each group of samples (ST, NASH and controls). These results
show that the Ct values obtained for both housekeeping genes (GADPH and ARP) were
very similar. Consequently, a media value of the 2 housekeeping Cts was used to
normalize the expression levels of the targeted genes (Figure 4).
Figure 3. mRNA expression ratio (Ct housekeeping / Ct target gene) of all the studied genes.
17
19. 32
Figure 4. Results of differential expression analysis obtained by delta-delta-Ct method .
The differential expression analysis showed that CYP2E1 has similar regulation
in controls and in ST, while it is up-regulated in NASH. We can also observe that ASNS,
STK11 and MTR are up-regulated in NASH. However, ASNS is down-regulated in ST, and
STK11 and MTR have similar expression in ST and controls. Additionally, SLC2A1 is
down-regulated in ST and NASH patients.
18
20. DISCUSSION
In association studies the possibility of false positive findings arises from a
combination of the characterization of small sample sizes, the poor description of case
and control samples, and the overestimation of the risks of genetic effects. In the
present study, the quality of the patient biological material was low and that is why
we got very small genotyping call rate of those samples, and therefore, the proportion
of the total genotyping success was very reduced (about 50%).
Among the obtained results, it is outstanding that the associated SNP located in
exon 9 of CYP2E1 (rs28969387) is a non-synonymous SNP that produces an
aminoacidic change (H457L). Structural changes or loss of function of this protein could
be related to this or other aminoacidic changes. We have seen that expression of
CYP2E1 is significantly up-regulated in NASH samples, however, it does not seem to be
altered at all in ST patients (see figure 3). Those results agree with previous studies 12
in which it is suggested that CYP2E1 could initiate oxidative stress leading to ST to
7, 12
NASH by production of reactive oxygen species (ROS) . Blocking CYP2E1 activity
prevents necroinflammatory changes in rats 4. One study by Jörn et al. 33
based in
overexpression of CYP2E1 in a hepatocyte cell line, reveals that increased CYP2E1
expression results in the down-regulation of insulin signaling, potentially contributing
to the insulin resistance associated with NAFLD. Factors related with NAFLD such as
ethanol exposure, a high-lipid and high-carbohydrate diet, fasting lost weigh and
diabetes, can increase CYP2E1 levels 4.
As shown in this study and in previous works 23, SLC2A1 is down-regulated in ST
and NASH patients (see figure 4). However, SLC2A1 has been found to be up-regulated
in obesity and diabetes patients 34, both diseases associated with NAFLD. The proper
understanding of this result could be very interesting since the oposite regulation of
SLC2A1 in ST/NASH compared to obesity/diabetes could suggest the potentiality of this
gene as a genetic marker to identify NAFLD.
4
When ST progresses to NASH, the insulin resistance occurs and in this
conditions, Cui et al.35 found over-expression of ASNS and, in addition, Sreekumar et
25
al. found general up-regulation in genes involved in protein synthesis in NASH
19
21. patients. Those data agree with our results, in which we found ASNS down-regulated in
ST while it is up-regulated in NASH (see figure 3).
36, 37
MTR enzyme has been shown down-regulated in alcoholic liver diseases
due probably to the direct effect of the alcohol in the regulation of this enzyme. In the
contrary, we saw that MTR was more expressed in the studied ST/NASH liver biopsies
than in the controls (Figure 3). This result could be understood if we consider that
NAFLD patients may drink from 0 to 40 gr of ethanol per week and we do not have this
information for the studied samples.
38
Nakau et al. suggest that lack of STK11 activity is a mechanism for HCC
38
development. In addition, STK11 plays a key role in the p53-dependent apoptosis ,
38
over-expression of STK11 in tumor cell lines results in cell cycle arrests and up-
regulation of STK11 could delay progression from NASH to HCC. What is more, STK11
phospholyration is related with an increase in expression of fatty acid synthase (FAS)
39
. Therefore, the up-regulation of this enzyme detected in the present study could be
the cause of an increasing lipid accumulation within hepatocytes in ST and NASH
patients.
It will be interesting to get the associated SNPs genotypes for the liver biopsies
where regulation of the corresponding genes was tested, in order to correlate
genotype to phenotype.
The obtained results open some new perspectives into the NAFLD research. It
will be required to describe the implications of the mentioned genes in the
pathogenesis of NAFLD, and hopefully, the resulting knowledge will even reveal some
new therapeutic targets.
20
22. CONCLUSIONS
The present project demonstrates for the first time that SLC2A1 is associated
with NAFLD, since 7 SNPs located within this gene showed significant association (p <
10-4).
Regulation of SLC2A1 in ST and NASH is opposite than in obese people. This
means that SLC2A1 could be a potential specific marker of NAFLD.
The regulation of the studied genes is always higher in NASH samples than in ST
ones. This could suggest that metabolism in NASH patients is more unbalanced than in
ST.
OUTLOOK
This project will continue first validating significant SNPs in other cohort of
case/control samples. Afterwards, functional studies of associated genes should be
carried out by either silencing down-regulated genes or over-expressing up-regulated
ones, followed by the analysis of the consequences of these regulations at genome-
wide level. The analysis of the transcriptome expression of the chosen in-vivo models
will be examined using high-throughput arrays.
Special efforts will be carried out to test that the regulation of associated genes
seen in liver biopsies is somehow reflected in the blood. This will be crucial to develop
non-invasive diagnostic tools.
AKNOWLEDGMENTS
This work was supported by a grant of Fundacion La caixa (obra social, number
BM06-227-0) coordinated by Mari Luz Martínez-Chantar. I would like to thank Ana M.
Aransay’s and Mari Luz Martinez-Chantar’s research groups for teaching me the
background, the hypothesis and the appropriate techniques that made this work
possible.
21
23. REFERENCES
1. Solis Herruzo JA, Garcia Ruiz I, Perez Carreras M, Munoz Yague MT. Non-
alcoholic fatty liver disease. From insulin resistance to mitochondrial
dysfunction. Rev Esp Enferm Dig 2006;98:844-74.
2. Osterreicher CH, Brenner DA. The genetics of nonalcoholic fatty liver disease.
Ann Hepatol 2007;6:83-8.
3. Duvnjak M, Lerotic I, Barsic N, Tomasic V, Virovic Jukic L, Velagic V.
Pathogenesis and management issues for non-alcoholic fatty liver disease.
World J Gastroenterol 2007;13:4539-50.
4. Chitturi S, Farrell GC. Etiopathogenesis of nonalcoholic steatohepatitis. Semin
Liver Dis 2001;21:27-41.
5. Clark JM, Brancati FL, Diehl AM. Nonalcoholic fatty liver disease.
Gastroenterology 2002;122:1649-57.
6. Nugent C, Younossi ZM. Evaluation and management of obesity-related
nonalcoholic fatty liver disease. Nat Clin Pract Gastroenterol Hepatol
2007;4:432-41.
7. Younossi ZM, Diehl AM, Ong JP. Nonalcoholic fatty liver disease: an agenda for
clinical research. Hepatology 2002;35:746-52.
8. Castera L. Non-invasive diagnosis of steatosis and fibrosis. Diabetes Metab
2008;34:674-9.
9. Angulo P, Keach JC, Batts KP, Lindor KD. Independent predictors of liver fibrosis
in patients with nonalcoholic steatohepatitis. Hepatology 1999;30:1356-62.
10. Fong DG, Nehra V, Lindor KD, Buchman AL. Metabolic and nutritional
considerations in nonalcoholic fatty liver. Hepatology 2000;32:3-10.
11. Shifflet A, Wu GY. Non-alcoholic Steatohepatitis: An Overview. J Formos Med
Assoc 2009;108:4-12.
12. Jou J, Choi SS, Diehl AM. Mechanisms of disease progression in nonalcoholic
fatty liver disease. Semin Liver Dis 2008;28:370-9.
13. Kim CH, Younossi ZM. Nonalcoholic fatty liver disease: a manifestation of the
metabolic syndrome. Cleve Clin J Med 2008;75:721-8.
22
24. 14. Pessayre D, Fromenty B. NASH: a mitochondrial disease. J Hepatol 2005;42:928-
40.
15. Bonkovsky HL, Jawaid Q, Tortorelli K, LeClair P, Cobb J, Lambrecht RW, Banner
BF. Non-alcoholic steatohepatitis and iron: increased prevalence of mutations
of the HFE gene in non-alcoholic steatohepatitis. J Hepatol 1999;31:421-9.
16. Erickson SK. Nonalcoholic fatty liver disease (NAFLD). J Lipid Res 2008.
17. Namikawa C, Shu-Ping Z, Vyselaar JR, Nozaki Y, Nemoto Y, Ono M, Akisawa N,
Saibara T, Hiroi M, Enzan H, Onishi S. Polymorphisms of microsomal triglyceride
transfer protein gene and manganese superoxide dismutase gene in non-
alcoholic steatohepatitis. J Hepatol 2004;40:781-6.
18. Tokushige K, Takakura M, Tsuchiya-Matsushita N, Taniai M, Hashimoto E,
Shiratori K. Influence of TNF gene polymorphisms in Japanese patients with
NASH and simple steatosis. J Hepatol 2007;46:1104-10.
19. Valenti L, Fracanzani AL, Dongiovanni P, Santorelli G, Branchi A, Taioli E, Fiorelli
G, Fargion S. Tumor necrosis factor alpha promoter polymorphisms and insulin
resistance in nonalcoholic fatty liver disease. Gastroenterology 2002;122:274-
80.
20. Song J, da Costa KA, Fischer LM, Kohlmeier M, Kwock L, Wang S, Zeisel SH.
Polymorphism of the PEMT gene and susceptibility to nonalcoholic fatty liver
disease (NAFLD). FASEB J 2005;19:1266-71.
21. Saffroy R, Pham P, Chiappini F, Gross-Goupil M, Castera L, Azoulay D, Barrier A,
Samuel D, Debuire B, Lemoine A. The MTHFR 677C > T polymorphism is
associated with an increased risk of hepatocellular carcinoma in patients with
alcoholic cirrhosis. Carcinogenesis 2004;25:1443-8.
22. Sazci A, Ergul E, Aygun C, Akpinar G, Senturk O, Hulagu S.
Methylenetetrahydrofolate reductase gene polymorphisms in patients with
nonalcoholic steatohepatitis (NASH). Cell Biochem Funct 2008;26:291-6.
23. Rubio A, Guruceaga E, Vazquez-Chantada M, Sandoval J, Martinez-Cruz LA,
Segura V, Sevilla JL, Podhorski A, Corrales FJ, Torres L, Rodriguez M, Aillet F, Ariz
U, Arrieta FM, Caballeria J, Martin-Duce A, Lu SC, Martinez-Chantar ML, Mato
JM. Identification of a gene-pathway associated with non-alcoholic
steatohepatitis. J Hepatol 2007;46:708-18.
23
25. 24. Westerbacka J, Kolak M, Kiviluoto T, Arkkila P, Siren J, Hamsten A, Fisher RM,
Yki-Jarvinen H. Genes involved in fatty acid partitioning and binding, lipolysis,
monocyte/macrophage recruitment, and inflammation are overexpressed in
the human fatty liver of insulin-resistant subjects. Diabetes 2007;56:2759-65.
25. Sreekumar R, Rosado B, Rasmussen D, Charlton M. Hepatic gene expression in
histologically progressive nonalcoholic steatohepatitis. Hepatology
2003;38:244-51.
26. Wang WY, Barratt BJ, Clayton DG, Todd JA. Genome-wide association studies:
theoretical and practical concerns. Nat Rev Genet 2005;6:109-18.
27. Zhao H, Pfeiffer R, Gail MH. Haplotype analysis in population genetics and
association studies. Pharmacogenomics 2003;4:171-8.
28. The International HapMap Project. Nature 2003;426:789-96.
29. Frazer KA, Ballinger DG, Cox DR, Hinds DA, Stuve LL, Gibbs RA, Belmont JW,
Boudreau A, Hardenbol P, Leal SM, Pasternak S, Wheeler DA, Willis TD, Yu F,
Yang H, Zeng C, Gao Y, Hu H, Hu W, Li C, Lin W, Liu S, Pan H, Tang X, Wang J,
Wang W, Yu J, Zhang B, Zhang Q, Zhao H, Zhou J, Gabriel SB, Barry R,
Blumenstiel B, Camargo A, Defelice M, Faggart M, Goyette M, Gupta S, Moore
J, Nguyen H, Onofrio RC, Parkin M, Roy J, Stahl E, Winchester E, Ziaugra L,
Altshuler D, Shen Y, Yao Z, Huang W, Chu X, He Y, Jin L, Liu Y, Sun W, Wang H,
Wang Y, Xiong X, Xu L, Waye MM, Tsui SK, Xue H, Wong JT, Galver LM, Fan JB,
Gunderson K, Murray SS, Oliphant AR, Chee MS, Montpetit A, Chagnon F,
Ferretti V, Leboeuf M, Olivier JF, Phillips MS, Roumy S, Sallee C, Verner A,
Hudson TJ, Kwok PY, Cai D, Koboldt DC, Miller RD, Pawlikowska L, Taillon-Miller
P, Xiao M, Tsui LC, Mak W, Song YQ, Tam PK, Nakamura Y, Kawaguchi T,
Kitamoto T, Morizono T, Nagashima A, Ohnishi Y, Sekine A, Tanaka T, Tsunoda
T, et al. A second generation human haplotype map of over 3.1 million SNPs.
Nature 2007;449:851-61.
30. de Bakker PI, Yelensky R, Pe'er I, Gabriel SB, Daly MJ, Altshuler D. Efficiency and
power in genetic association studies. Nat Genet 2005;37:1217-23.
31. Gabriel SB, Schaffner SF, Nguyen H, Moore JM, Roy J, Blumenstiel B, Higgins J,
DeFelice M, Lochner A, Faggart M, Liu-Cordero SN, Rotimi C, Adeyemo A,
24
26. Cooper R, Ward R, Lander ES, Daly MJ, Altshuler D. The structure of haplotype
blocks in the human genome. Science 2002;296:2225-9.
32. Giulietti A, Overbergh L, Valckx D, Decallonne B, Bouillon R, Mathieu C. An
overview of real-time quantitative PCR: applications to quantify cytokine gene
expression. Methods 2001;25:386-401.
33. Schattenberg JM, Wang Y, Singh R, Rigoli RM, Czaja MJ. Hepatocyte CYP2E1
overexpression and steatohepatitis lead to impaired hepatic insulin signaling. J
Biol Chem 2005;280:9887-94.
34. MacLaren R, Cui W, Simard S, Cianflone K. Influence of obesity and insulin
sensitivity on insulin signaling genes in human omental and subcutaneous
adipose tissue. J Lipid Res 2008;49:308-23.
35. Cui H, Darmanin S, Natsuisaka M, Kondo T, Asaka M, Shindoh M, Higashino F,
Hamuro J, Okada F, Kobayashi M, Nakagawa K, Koide H. Enhanced expression
of asparagine synthetase under glucose-deprived conditions protects
pancreatic cancer cells from apoptosis induced by glucose deprivation and
cisplatin. Cancer Res 2007;67:3345-55.
36. Barak AJ, Beckenhauer HC, Kharbanda KK, Tuma DJ. Chronic ethanol
consumption increases homocysteine accumulation in hepatocytes. Alcohol
2001;25:77-81.
37. Kerai MD, Waterfield CJ, Kenyon SH, Asker DS, Timbrell JA. Reversal of ethanol-
induced hepatic steatosis and lipid peroxidation by taurine: a study in rats.
Alcohol Alcohol 1999;34:529-41.
38. Nakau M, Miyoshi H, Seldin MF, Imamura M, Oshima M, Taketo MM.
Hepatocellular carcinoma caused by loss of heterozygosity in Lkb1 gene
knockout mice. Cancer Res 2002;62:4549-53.
39. Hou X, Xu S, Maitland-Toolan KA, Sato K, Jiang B, Ido Y, Lan F, Walsh K,
Wierzbicki M, Verbeuren TJ, Cohen RA, Zang M. SIRT1 regulates hepatocyte
lipid metabolism through activating AMP-activated protein kinase. J Biol Chem
2008;283:20015-26.
25