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��ࡱ�>��	AD����>?@�����������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������������[�	��0�'bjbj����	40ΐΐϺ+�������WWWWW����kkk8�D�Lk�D�3�1!G!G!G!6"��"D*#$�C�C�C�C�C�C�C�G�3J>�C�W4&6"6"4&4&�CWWG!G!�oD�1�1�14&�WG!WG!?��14&�C�1�1x3�3G!����@KY���k+��3�>�D0�D�3,qJ-�qJ�3�3�qJWb9�N#h�#J�1$<<$�N#N#N#�C�C�1N#N#N#�D4&4&4&4&��������������������������������������������������������������������qJN#N#N#N#N#N#N#N#N#	:	Analysis of Methylation and Expression Profiles of CPT1-A and APOE Genes in Patients with NAFLD 

1Dor Mohammad Kordi-Tamandani1*, Mohammad Hashemi2, Tayebe Baranzehi 

1Departement of biology, university of Sistan and Baluchestan, Zahedan, Iran

2Department of clinical biochemistry, Zahedan university of Medical Sciences, Zahedan, Iran

*Corresponding Authors:
Dr D.M. Kordi Tamandani and Dr Adam Torkamanzehi 
Department of Biology, 
University of Sistan and Baluchestan, 
P.O.Box 98155-987, Zahedan, Iran. 
Tell: +98-541-2452335, Fax: +98-541-2446565















Abstract
Non-alcoholic fatty liver disease (NAFLD) is the most common cause of abnormal hepatic steatosis in the absence of alcohol abuse worldwide. The aim of this study was to investigate the effect of promoter methylation of carnitine palmitoyltransferase I (CPT1) and apolipoprotein E  (APOE) genes on the risk of non-alcoholic fatty liver disease (NAFLD).
METHODS: The promoter methylation of APOE and CPT-1A were analyzed using a methylation-specific polymerase chain reaction (MS-PCR) in blood samples taken from 80 NAFLD individuals and 100 healthy controls. The expression levels of APOE and CPT-1A were also assessed in 10 blood mRNA samples from NAFLD patients. These cases were compared to the blood samples of healthy controls(n=10) with real-time quantitative reverse transcriptase PCR. 
RESULTS: The percentage of methylation for CPT-1A gene was significant between normal individuals and patients. The APOE gene methylation was not significant between cases and controls. A statistically significant relationship was found for methylation of CPT-1A between cases and controls (p<0.001). The relative expression of CPT-1A and APOE mRNA in NAFLD blood was no significantly different in comparison of blood samples between healthy controls and cases. 
CONCLUSION: The present outcomes indicate that the methylation status of the CPT-1A gene has a significant function in the process of NAFLD and suggest further study with a large sample size for this reason.

Keywords: CPT-1A; APOE; gene; methylation; expression; NAFLD









Introduction 
Nonalcoholic fatty liver disease (NAFLD) is a vast term that refers to a wide range of lesions that vary from steatosis without necro-inflammatory injury to active lesions of hepatocyte injury, inflammation and cell death  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]1,2[. The etiology of the syndrome is unknown, but it is reported to be linked with the excess accumulation of adipose tissue, which is associated with a large number of metabolic disorders such as cardiovascular disease, type 2 diabetes, obesity and hyperlipidemia  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]3[. The prevalence of NAFLD in the general population depends on the type of screening test, varying from 2.8% to 46% in the unselected population worldwide  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]4[. In addition to a higher prevalence of NAFLD in patients with the above mentioned disorders, it can also be induced by a number of genetic variations  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]5[. Environmental and genetic factors play an important role in etiology of obesity as a multi-factorial disorder  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]6[. Finding the interaction of genetic and environmental factors plays a critical role in prevention and treatment of diseases. The present study focuses on analysis of epigenetic variation (DNA methylation), relative to which, until now, there has been no published data regarding its effect on the progress of NALFD. The genes involved in fatty acid metabolism can be considered as good candidate genes to explain such variations  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]7[ . One of these genes�the candidate for the current study�is the gene coding for carnitine palmitoyltransferase I (CPT1); it  has  two isoforms: CPT1A is located on  chromosome 11q13.1�q13.2 and is mainly expressed in the liver; whereas CPT1B is mapped on chromosome 22q13.33 and is mostly found in skeletal muscle and heart, but also in adipose tissue and the testes  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]8[. It has an essential role in �- oxidation of long-chain fatty acids in the mitochondria  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]9, ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>10[. Another studied gene was the apolipoprotein E, which  is situated on chromosome 19; its protein product (APOE) has three general isoforms�E2, E3 and E4�encoded by the alleles �2, �3 and �4, of which the �3/� 3 genotype takes place  in about one-half to two-thirds of people in the majority populations. The three regular isoforms of the protein link in a different way with specific lipoprotein receptors, finally changing circulating levels of cholesterol during different portions of the lipoprotein metabolism pathway, such as hepatic binding, uptake, catabolism of chylomicrons, high density lipoprotein and very low density lipoprotein formation. Indeed, it has been suggested as a main regulator of blood lipid levels in humans  ADDIN EN.CITE  ADDIN EN.CITE.DATA ]11,12[
Zhang et al. (2001) have shown that the E4 allele of APOE was significantly linked with increased plasma lipid levels and  body mass index (BMI) in obese children  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]13[.The aim of this study was to examine the association between APOE and CPT-1A gene methylation and their expression profiles in southeast Iranian patients with NAFLD.
Materials and Methods
Study subjects and specimens 
Eighty NAFLD samples and the samples of 100 healthy individuals were obtained from Ali-ibn Abi Talib Hospital, Zahedan, between 2009 and 2010. The ethical committee at the Medical University of Zahedan approved this study. The lymphocytes (or leukocytes) in peripheral blood samples (normal and patient) were collected as previously described  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]14[ briefly. Detection of NAFLD was performed by clinical assessment, and laboratory and sonographic results, given that the patients had not given permission to undertake liver biopsy. Healthy individuals were selected from those in the Zahedan population who had participated in a project pertaining to metabolic syndrome and who had normal status for blood glucose, blood pressure, lipid profile, waist circumference, BMI, and no history of systematic disease Table1.
DNA extraction and methylation-specific PCR analysis
Two ml of collected blood per sample was used for extraction of genomic DNA by the phenol-chloroform method, of which 2 �g were used for bisulfate treatment, as described elsewhere  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]14[. The sequence and annealing temperature of primers of MS-PCR are given in Table 2. MSP was carried out in a total volume of 25 �L, containing 1.5 �L bisulfate modified DN , 20mM Tris-HCl (pH 8.4), 50mM KCl, 0.2 mM each dNTP, 1.5 mM MgCl2, 1 pM each primer, and 2.5 units of hot-start Taq (Fermentas,USA). Positive controls (in vitro methylated and bisulfite treated human placental DNA) and negative controls (no template DNA) were included in all reactions. PCR products were assayed in a 2% agarose gel RNA extraction and reverse transcription�polymerase chain reaction one ml of fresh blood sample was used for RNA extraction by high pure RNA Isolation Kit (Roche, Cat. No. 11 828 665 001) according to manufacturer s instructions. 
	The scan drop spectrophotometer was used for detection of concentration and purity of isolated RNA. Total RNA (1�g) was reverse-transcribed with RevertAid-First-Strand cDNA Synthesis Kit (Fermentas, Cat. No. K1621;) in a final volume of 20 �L. The sequences of primers and amplification conditions are listed in Table 2 .
APOE and CTP1 mRNA expression levels were estimated by real-time quantitative  PCR using the AB15700 (Applied Biosystems). Samples were tested in a 2o�L reaction mixture containing 1�L of cDNA, 1�L of each specific primer, 10�L of 2� Sybergreen and 8�L of  RNase free water. RNA18s was used as an internal control for normalizing of the APOE and CPT1 expression levels. Assessment of gene expression was performed with (2-��CT) methods as previously described  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]15[.

Statistical Analysis
Statistical correlation was tested by t- , chi-square and Mann-Witney tests, with SPSS Version 17.0 (SPSS Inc.; Chicago, IL); a significance p-value d" 0.05 was set.

Results	
CPT1-A and APOE gene methylation 
The results of CPT1 and APOE gene methylation for both cases and controls are shown in Table 3. As indicated, the APOE gene was 38.75% methylated in NAFLD patients and 32% methylated in healthy individuals. This variation is not significant between healthy controls and patients. A statistically significant relationship (p<0.001) was established for the CTP1-A gene, which was 85.0% methylated in normal individuals and 56% methylated in NAFLD individuals.

CPT1-A and APOE gene expression
The differences between CPT1-A and APOE mRNA levels were tested in 10 normal blood samples and 10 blood samples were taken from NAFLD patients. As shown in Tables 4, there was no statistically significant association for relative gene expression in a comparison of blood samples between cases and controls for the APOE and CPT1-A genes. Additional studies utilizing larger sample sizes are necessary to achieve a conclusion regarding the effect of altered gene expression of CPT1-A and APOE genes in NAFLD.

Discussion 
Hypermethylation of CpG islands is now a well-established epigenetic alteration in the development and progression of many diseases as a cause of transcriptional silencing  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]16[. The detection of altered DNA methylation could be used for the finding of malignant transformed cells  ADDIN EN.CITE  ADDIN EN.CITE.DATA ]17, 16, 18, 19[, and might be used as an idea for potential molecular diagnosis in the not-distant future in the field of clinical medicine. The precise role of DNA methylation in the development of NAFLD remains unclear.Therfore, We have begun work on analysis of promoter DNA methylation on many candidate genes that have critical roles in methylation of lipid metabolism, which might be linked with NAFLD  ADDIN EN.CITE <EndNote><Cite><Author>Greenfield</Author><Year>2008</Year><RecNum>38</RecNum><DisplayText>(8)</DisplayText><record><rec-number>38</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">38</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Greenfield, V.</author><author>Cheung, O.</author><author>Sanyal, A. J.</author></authors></contributors><auth-address>Divisoipn of Gastroenterology, Hepatology and Nutrition, Department of Internal Medicine, Virginia Commonwealth University School of Medicine, Richmond, Virginia, USA.</auth-address><titles><title>Recent advances in nonalcholic fatty liver disease</title><secondary-title>Curr Opin Gastroenterol</secondary-title></titles><periodical><full-title>Curr Opin Gastroenterol</full-title></periodical><pages>320-7</pages><volume>24</volume><number>3</number><edition>2008/04/15</edition><keywords><keyword>Fatty Liver/diagnosis/*physiopathology/therapy</keyword><keyword>Hepatocytes/pathology</keyword><keyword>Humans</keyword><keyword>Inflammation/physiopathology</keyword><keyword>Insulin Resistance/physiology</keyword><keyword>Liver Cirrhosis/physiopathology</keyword></keywords><dates><year>2008</year><pub-dates><date>May</date></pub-dates></dates><isbn>1531-7056 (Electronic)&#xD;0267-1379 (Linking)</isbn><accession-num>18408460</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=18408460</url></related-urls></urls><electronic-resource-num>10.1097/MOG.0b013e3282fbccf2&#xD;00001574-200805000-00009 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>]20[. Dysregulation of these gene expressions through interaction with environmental factors plays a critical role in development of disease., regarding methylation�s effect on the progress of NAFLD. Hence, such a study breaks ground for doing more research in this field, which may ultimately lead to finding new target genes and could have significance in molecular diagnosis of NAFLD. 
The APOE gene provides instructions for making a protein called apolipoprotein E, which is responsible for packaging cholesterol and other fats and carrying them through the bloodstream in the form of low-density lipoproteins (VLDLs)  ADDIN EN.CITE <EndNote><Cite><Author>Carter</Author><Year>2007</Year><RecNum>45</RecNum><DisplayText>(3)</DisplayText><record><rec-number>45</rec-number><foreign-keys><key app="EN" db-id="a5tar9rpatdv2gez2rlppr0fd99tvw90zd22">45</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Carter, C. J.</author></authors></contributors><auth-address>chris_car@yahoo.com</auth-address><titles><title>Convergence of genes implicated in Alzheimer&apos;s disease on the cerebral cholesterol shuttle: APP, cholesterol, lipoproteins, and atherosclerosis</title><secondary-title>Neurochem Int</secondary-title></titles><periodical><full-title>Neurochem Int</full-title></periodical><pages>12-38</pages><volume>50</volume><number>1</number><edition>2006/09/16</edition><keywords><keyword>Alzheimer Disease/*genetics</keyword><keyword>Amyloid beta-Protein Precursor/*metabolism</keyword><keyword>Atherosclerosis/*metabolism</keyword><keyword>Biological Transport</keyword><keyword>Brain/*metabolism</keyword><keyword>Cholesterol/*metabolism</keyword><keyword>Homeostasis</keyword><keyword>Humans</keyword><keyword>Lipoproteins/*metabolism</keyword></keywords><dates><year>2007</year><pub-dates><date>Jan</date></pub-dates></dates><isbn>0197-0186 (Print)&#xD;0197-0186 (Linking)</isbn><accession-num>16973241</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=16973241</url></related-urls></urls><electronic-resource-num>S0197-0186(06)00247-6 [pii]&#xD;10.1016/j.neuint.2006.07.007</electronic-resource-num><language>eng</language></record></Cite></EndNote>]21[. Data are very limited regarding APOE. Sozia et al. (2008) have reported that the polymorphism of APOE (E allele) is significantly linked with NASH  ADDIN EN.CITE  ADDIN EN.CITE.DATA ]22[. Also, a significant correlation was found between concentrations of APOE plasma levels and pathogenicity of fatty liver disease  ADDIN EN.CITE  ADDIN EN.CITE.DATA ]23[ .  


Certain studies have suggested that the relationship between APOE polymorphism and coronary heart disease (CHD) cannot be explained by a single factor, and that multiple factors may be involved in its nature[24]. 
With respect to biomarkers, APOE epsilon4 is not confirmed as a cerebrospinal fluid biomarker signature, though it is similarly observed in Alzheimer's disease as an important genetic marker  ADDIN EN.CITE  ADDIN EN.CITE.DATA ]25, 26[. CPT1A has a significant role in transporting fatty acids to mitochondria for �-oxidation  ADDIN EN.CITE  ADDIN EN.CITE.DATA ]27[. 
	An animal model study has shown that the CPT-1A mRNA levels are higher in livers of animals treated with 3-Thia drug  ADDIN EN.CITE  ADDIN EN.CITE.DATA ]28[. Shen et al.�s (2006) data showed that abnormal CpG island methylation of CPT1A takes place during the differentiation of human embryonic stem cells (hESC)  ADDIN EN.CITE  ADDIN EN.CITE.DATA ]29[. In the present study, we have found a significant status of CPT1-A gene methylation between NAFLD patients and healthy individuals. Considering the role of CPT1-A in metabolism of fatty acids, logically, we expected high methylation frequency (85%) in healthy individuals, but in NAFLD patients with high levels of fatty acids, in cases of low expression of CPT1- A, which might have been caused by hypermethylation of CPG islands of CPT1-A gene. 
This study the first, to the best of our knowledge, to focus on analysis of methylation of susceptible genes in NAFLD patients, and the way in which it highlights the significant change of the CTP1-A gene as a cause in interaction with environmental factors. Given the limitations of this research, further collaboration using a focused study design on larger population samples is necessary for verification and understanding of the genetic and epigenetic etiology of NAFLD, which it is still a long way off.
Acknowledgments
The present authors are especially grateful to all the patients and controls who took part in this study. Financial support was provided by the University of Sistan and Baluchestan, and the Zahedan University of Medical Sciences in Iran.

References
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C.</author></authors></contributors><auth-address>Department of Biostatistics and Epidemiology, College of Public Health, University of Oklahoma, Oklahoma City 73190, OK. june-eichner@ouhsc.edu</auth-address><titles><title>Apolipoprotein E polymorphism and cardiovascular disease: a HuGE review</title><secondary-title>Am J Epidemiol</secondary-title></titles><periodical><full-title>Am J Epidemiol</full-title></periodical><pages>487-95</pages><volume>155</volume><number>6</number><edition>2002/03/08</edition><keywords><keyword>Alleles</keyword><keyword>Animals</keyword><keyword>Apolipoproteins E/blood/*genetics</keyword><keyword>Cardiovascular Diseases/blood/*genetics</keyword><keyword>Cholesterol/blood</keyword><keyword>Chromosomes, Human, Pair 19/genetics</keyword><keyword>Gene Frequency</keyword><keyword>Genetic Variation</keyword><keyword>Genetics, Population</keyword><keyword>Genotype</keyword><keyword>Humans</keyword><keyword>Lipoproteins/*blood</keyword><keyword>Phenotype</keyword><keyword>*Polymorphism, Genetic</keyword><keyword>Risk Factors</keyword></keywords><dates><year>2002</year><pub-dates><date>Mar 15</date></pub-dates></dates><isbn>0002-9262 (Print)&#xD;0002-9262 (Linking)</isbn><accession-num>11882522</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=11882522</url></related-urls></urls><language>eng</language></record></Cite><Cite><Author>Bazzaz</Author><Year>2010</Year><RecNum>9</RecNum><record><rec-number>9</rec-number><foreign-keys><key app="EN" db-id="a5tar9rpatdv2gez2rlppr0fd99tvw90zd22">9</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Bazzaz, J. T.</author><author>Nazari, M.</author><author>Nazem, H.</author><author>Amiri, P.</author><author>Fakhrzadeh, H.</author><author>Heshmat, R.</author><author>Abbaszadeh, S.</author><author>Amoli, M. 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A.</author></authors></contributors><auth-address>University of Southern California/Norris Comprehensive Cancer Center, Department of Biochemistry and Molecular Biology, Keck School of Medicine, University of Southern California, Los Angeles, California 90089-9181, USA. Martin.Widschwendter@uibk.ac.at</auth-address><titles><title>The potential prognostic, predictive, and therapeutic values of DNA methylation in cancer. Commentary re: J. Kwong et al., Promoter hypermethylation of multiple genes in nasopharyngeal carcinoma. Clin. Cancer Res., 8: 131-137, 2002, and H-Z. Zou et al., Detection of aberrant p16 methylation in the serum of colorectal cancer patients. Clin. 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W.</author><author>To, K. F.</author><author>Teo, P. M.</author><author>Johnson, P. J.</author><author>Huang, D. P.</author></authors></contributors><auth-address>Department of Anatomical and Cellular Pathology, The Chinese University of Hong Kong, Hong Kong SAR, People&apos;s Republic of China. s993101@mailserv.cuhk.edu.hk</auth-address><titles><title>Promoter hypermethylation of multiple genes in nasopharyngeal carcinoma</title><secondary-title>Clin Cancer Res</secondary-title></titles><periodical><full-title>Clin Cancer Res</full-title></periodical><pages>131-7</pages><volume>8</volume><number>1</number><edition>2002/01/22</edition><keywords><keyword>Apoptosis Regulatory Proteins</keyword><keyword>Calcium-Calmodulin-Dependent Protein Kinases/genetics</keyword><keyword>*Cell Cycle Proteins</keyword><keyword>Cyclin-Dependent Kinase Inhibitor p15</keyword><keyword>Cyclin-Dependent Kinase Inhibitor p16/genetics</keyword><keyword>*DNA Methylation</keyword><keyword>DNA Primers/chemistry</keyword><keyword>Gene Expression</keyword><keyword>*Genes, Tumor Suppressor</keyword><keyword>Glutathione S-Transferase pi</keyword><keyword>Glutathione Transferase/genetics</keyword><keyword>Helminth Proteins/genetics</keyword><keyword>Humans</keyword><keyword>Isoenzymes/genetics</keyword><keyword>Muscle Proteins/genetics</keyword><keyword>Mutation</keyword><keyword>Nasopharyngeal Neoplasms/*genetics/metabolism</keyword><keyword>Neoplasm Proteins/*genetics/metabolism</keyword><keyword>Neoplasm Recurrence, Local/*genetics/mortality/pathology</keyword><keyword>O(6)-Methylguanine-DNA Methyltransferase/genetics</keyword><keyword>Polymerase Chain Reaction</keyword><keyword>Prognosis</keyword><keyword>Promoter Regions, Genetic/*genetics</keyword><keyword>Receptors, Retinoic Acid/genetics</keyword><keyword>Survival Rate</keyword><keyword>Transcription Factors/genetics</keyword><keyword>Transplantation, Heterologous</keyword><keyword>Tumor Cells, Cultured</keyword><keyword>*Tumor Suppressor Proteins</keyword></keywords><dates><year>2002</year><pub-dates><date>Jan</date></pub-dates></dates><isbn>1078-0432 (Print)&#xD;1078-0432 (Linking)</isbn><accession-num>11801549</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=11801549</url></related-urls></urls><language>eng</language></record></Cite><Cite><Author>Zou</Author><Year>2002</Year><RecNum>179</RecNum><record><rec-number>179</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">179</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Zou, H.</author><author>Yu, B.</author><author>Zhao, R.</author><author>Wang, Z.</author><author>Cang, H.</author><author>Li, D.</author><author>Feng, G.</author><author>Yi, J.</author></authors></contributors><auth-address>Department of General Surgery, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China.</auth-address><titles><title>Detection of aberrant p16 methylation in the serum of colorectal cancer patients</title><secondary-title>Zhonghua Yu Fang Yi Xue Za Zhi</secondary-title></titles><periodical><full-title>Zhonghua Yu Fang Yi Xue Za Zhi</full-title></periodical><pages>499-501</pages><volume>36</volume><number>7</number><edition>2002/11/02</edition><dates><year>2002</year><pub-dates><date>Dec</date></pub-dates></dates><isbn>0253-9624 (Print)&#xD;0253-9624 (Linking)</isbn><accession-num>12411153</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12411153</url></related-urls></urls><language>eng</language></record></Cite><Cite><Author>Wajed</Author><Year>2001</Year><RecNum>183</RecNum><record><rec-number>183</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">183</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Wajed, S. A.</author><author>Laird, P. W.</author><author>DeMeester, T. R.</author></authors></contributors><auth-address>Department of Surgery, University of Southern California School of Medicine, Los Angeles, California 90033, USA.</auth-address><titles><title>DNA methylation: an alternative pathway to cancer</title><secondary-title>Ann Surg</secondary-title></titles><periodical><full-title>Ann Surg</full-title></periodical><pages>10-20</pages><volume>234</volume><number>1</number><edition>2001/06/23</edition><keywords><keyword>Breast Neoplasms/genetics/physiopathology</keyword><keyword>Colonic Neoplasms/genetics/physiopathology</keyword><keyword>*DNA Methylation</keyword><keyword>Female</keyword><keyword>Gene Expression Regulation, Neoplastic</keyword><keyword>Gene Silencing</keyword><keyword>Humans</keyword><keyword>Microsatellite Repeats</keyword><keyword>Neoplasms/genetics/*physiopathology/therapy</keyword><keyword>Ovarian Neoplasms/genetics/physiopathology</keyword><keyword>Point Mutation</keyword><keyword>Promoter Regions, Genetic</keyword></keywords><dates><year>2001</year><pub-dates><date>Jul</date></pub-dates></dates><isbn>0003-4932 (Print)&#xD;0003-4932 (Linking)</isbn><accession-num>11420478</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=11420478</url></related-urls></urls><custom2>1421942</custom2><language>eng</language></record></Cite></EndNote>�D<EndNote><Cite><Author>Widschwendter</Author><Year>2002</Year><RecNum>184</RecNum><DisplayText>(24, 11, 28, 23)</DisplayText><record><rec-number>184</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">184</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Widschwendter, M.</author><author>Jones, P. A.</author></authors></contributors><auth-address>University of Southern California/Norris Comprehensive Cancer Center, Department of Biochemistry and Molecular Biology, Keck School of Medicine, University of Southern California, Los Angeles, California 90089-9181, USA. Martin.Widschwendter@uibk.ac.at</auth-address><titles><title>The potential prognostic, predictive, and therapeutic values of DNA methylation in cancer. Commentary re: J. Kwong et al., Promoter hypermethylation of multiple genes in nasopharyngeal carcinoma. Clin. Cancer Res., 8: 131-137, 2002, and H-Z. Zou et al., Detection of aberrant p16 methylation in the serum of colorectal cancer patients. Clin. Cancer Res., 8: 188-191, 2002</title><secondary-title>Clin Cancer Res</secondary-title></titles><periodical><full-title>Clin Cancer Res</full-title></periodical><pages>17-21</pages><volume>8</volume><number>1</number><edition>2002/01/22</edition><keywords><keyword>Chromosome Aberrations</keyword><keyword>Cyclin-Dependent Kinase Inhibitor p16/genetics</keyword><keyword>*DNA Methylation</keyword><keyword>DNA, Neoplasm/blood/*genetics</keyword><keyword>Gene Expression</keyword><keyword>Gene Silencing</keyword><keyword>Humans</keyword><keyword>Microsatellite Repeats</keyword><keyword>Mutation</keyword><keyword>Neoplasms/diagnosis/*genetics/mortality</keyword><keyword>Predictive Value of Tests</keyword><keyword>Prognosis</keyword><keyword>Promoter Regions, Genetic</keyword><keyword>Survival Rate</keyword></keywords><dates><year>2002</year><pub-dates><date>Jan</date></pub-dates></dates><isbn>1078-0432 (Print)&#xD;1078-0432 (Linking)</isbn><accession-num>11801535</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=11801535</url></related-urls></urls><language>eng</language></record></Cite><Cite><Author>Kwong</Author><Year>2002</Year><RecNum>176</RecNum><record><rec-number>176</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">176</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Kwong, J.</author><author>Lo, K. W.</author><author>To, K. F.</author><author>Teo, P. M.</author><author>Johnson, P. J.</author><author>Huang, D. P.</author></authors></contributors><auth-address>Department of Anatomical and Cellular Pathology, The Chinese University of Hong Kong, Hong Kong SAR, People&apos;s Republic of China. s993101@mailserv.cuhk.edu.hk</auth-address><titles><title>Promoter hypermethylation of multiple genes in nasopharyngeal carcinoma</title><secondary-title>Clin Cancer Res</secondary-title></titles><periodical><full-title>Clin Cancer Res</full-title></periodical><pages>131-7</pages><volume>8</volume><number>1</number><edition>2002/01/22</edition><keywords><keyword>Apoptosis Regulatory Proteins</keyword><keyword>Calcium-Calmodulin-Dependent Protein Kinases/genetics</keyword><keyword>*Cell Cycle Proteins</keyword><keyword>Cyclin-Dependent Kinase Inhibitor p15</keyword><keyword>Cyclin-Dependent Kinase Inhibitor p16/genetics</keyword><keyword>*DNA Methylation</keyword><keyword>DNA Primers/chemistry</keyword><keyword>Gene Expression</keyword><keyword>*Genes, Tumor Suppressor</keyword><keyword>Glutathione S-Transferase pi</keyword><keyword>Glutathione Transferase/genetics</keyword><keyword>Helminth Proteins/genetics</keyword><keyword>Humans</keyword><keyword>Isoenzymes/genetics</keyword><keyword>Muscle Proteins/genetics</keyword><keyword>Mutation</keyword><keyword>Nasopharyngeal Neoplasms/*genetics/metabolism</keyword><keyword>Neoplasm Proteins/*genetics/metabolism</keyword><keyword>Neoplasm Recurrence, Local/*genetics/mortality/pathology</keyword><keyword>O(6)-Methylguanine-DNA Methyltransferase/genetics</keyword><keyword>Polymerase Chain Reaction</keyword><keyword>Prognosis</keyword><keyword>Promoter Regions, Genetic/*genetics</keyword><keyword>Receptors, Retinoic Acid/genetics</keyword><keyword>Survival Rate</keyword><keyword>Transcription Factors/genetics</keyword><keyword>Transplantation, Heterologous</keyword><keyword>Tumor Cells, Cultured</keyword><keyword>*Tumor Suppressor Proteins</keyword></keywords><dates><year>2002</year><pub-dates><date>Jan</date></pub-dates></dates><isbn>1078-0432 (Print)&#xD;1078-0432 (Linking)</isbn><accession-num>11801549</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=11801549</url></related-urls></urls><language>eng</language></record></Cite><Cite><Author>Zou</Author><Year>2002</Year><RecNum>179</RecNum><record><rec-number>179</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">179</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Zou, H.</author><author>Yu, B.</author><author>Zhao, R.</author><author>Wang, Z.</author><author>Cang, H.</author><author>Li, D.</author><author>Feng, G.</author><author>Yi, J.</author></authors></contributors><auth-address>Department of General Surgery, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China.</auth-address><titles><title>Detection of aberrant p16 methylation in the serum of colorectal cancer patients</title><secondary-title>Zhonghua Yu Fang Yi Xue Za Zhi</secondary-title></titles><periodical><full-title>Zhonghua Yu Fang Yi Xue Za Zhi</full-title></periodical><pages>499-501</pages><volume>36</volume><number>7</number><edition>2002/11/02</edition><dates><year>2002</year><pub-dates><date>Dec</date></pub-dates></dates><isbn>0253-9624 (Print)&#xD;0253-9624 (Linking)</isbn><accession-num>12411153</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=12411153</url></related-urls></urls><language>eng</language></record></Cite><Cite><Author>Wajed</Author><Year>2001</Year><RecNum>183</RecNum><record><rec-number>183</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">183</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Wajed, S. 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W.</author></authors></contributors><auth-address>Center for Health Promotion, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.</auth-address><titles><title>The relationship between apolipoprotein E polymorphism, lipoprotein (a) and fatty liver disease</title><secondary-title>Hepatogastroenterology</secondary-title></titles><periodical><full-title>Hepatogastroenterology</full-title></periodical><pages>1832-5</pages><volume>52</volume><number>66</number><edition>2005/12/13</edition><keywords><keyword>Adult</keyword><keyword>Age Distribution</keyword><keyword>Apolipoproteins E/genetics/*metabolism</keyword><keyword>Case-Control Studies</keyword><keyword>Cross-Sectional Studies</keyword><keyword>Disease Progression</keyword><keyword>Fatty Liver/epidemiology/*genetics/pathology</keyword><keyword>Female</keyword><keyword>Gene Expression Regulation</keyword><keyword>Genetic Markers/genetics</keyword><keyword>Humans</keyword><keyword>Lipoprotein(a)/genetics/*metabolism</keyword><keyword>Liver Function Tests</keyword><keyword>Logistic Models</keyword><keyword>Male</keyword><keyword>Middle Aged</keyword><keyword>Odds Ratio</keyword><keyword>*Polymorphism, Genetic</keyword><keyword>Probability</keyword><keyword>Prognosis</keyword><keyword>Reference Values</keyword><keyword>Risk Assessment</keyword><keyword>Sensitivity and Specificity</keyword><keyword>Severity of Illness Index</keyword><keyword>Sex Distribution</keyword></keywords><dates><year>2005</year><pub-dates><date>Nov-Dec</date></pub-dates></dates><isbn>0172-6390 (Print)&#xD;0172-6390 (Linking)</isbn><accession-num>16334787</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=16334787</url></related-urls></urls><language>eng</language></record></Cite></EndNote>�
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K.</author></authors></contributors><auth-address>Department of Laboratory Medicine, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India.</auth-address><titles><title>IL-6-174 G/C and ApoE gene polymorphisms in Alzheimer&apos;s and vascular dementia patients attending the cognitive disorder clinic of the All India Institute of Medical Sciences, New Delhi</title><secondary-title>Dement Geriatr Cogn Disord</secondary-title></titles><periodical><full-title>Dement Geriatr Cogn Disord</full-title></periodical><pages>461-8</pages><volume>30</volume><number>6</number><edition>2011/01/22</edition><dates><year>2010</year></dates><isbn>1421-9824 (Electronic)&#xD;1420-8008 (Linking)</isbn><accession-num>21252539</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=21252539</url></related-urls></urls><electronic-resource-num>000321666 [pii]&#xD;10.1159/000321666</electronic-resource-num><language>eng</language></record></Cite><Cite><Author>Zetzsche</Author><Year>2010</Year><RecNum>44</RecNum><record><rec-number>44</rec-number><foreign-keys><key app="EN" db-id="a5tar9rpatdv2gez2rlppr0fd99tvw90zd22">44</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Zetzsche, T.</author><author>Rujescu, D.</author><author>Hardy, J.</author><author>Hampel, H.</author></authors></contributors><auth-address>Department of Psychiatry and Psychotherapy, Ludwig-Maximilian-University, Nussbaumstrasse 7, Munich, Germany. thomas.zetzsche@ med.uni-muenchen.de</auth-address><titles><title>Advances and perspectives from genetic research: development of biological markers in Alzheimer&apos;s disease</title><secondary-title>Expert Rev Mol Diagn</secondary-title></titles><periodical><full-title>Expert Rev Mol Diagn</full-title></periodical><pages>667-90</pages><volume>10</volume><number>5</number><edition>2010/07/16</edition><keywords><keyword>Alzheimer Disease/*genetics/metabolism/physiopathology/therapy</keyword><keyword>Apolipoproteins E/genetics</keyword><keyword>Biological Markers/*metabolism</keyword><keyword>Epigenesis, Genetic</keyword><keyword>Gene Therapy</keyword><keyword>*Genetic Predisposition to Disease</keyword><keyword>Genetic Variation</keyword><keyword>Humans</keyword><keyword>Mutation</keyword><keyword>Pharmacogenetics</keyword><keyword>Phenotype</keyword><keyword>RNA Interference</keyword></keywords><dates><year>2010</year><pub-dates><date>Jul</date></pub-dates></dates><isbn>1744-8352 (Electronic)&#xD;1473-7159 (Linking)</isbn><accession-num>20629514</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=20629514</url></related-urls></urls><electronic-resource-num>10.1586/erm.10.48</electronic-resource-num><language>eng</language></record></Cite></EndNote>	D<EndNote><Cite><Author>Rajakumar</Author><Year>2009</Year><RecNum>127</RecNum><DisplayText>(15)</DisplayText><record><rec-number>127</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">127</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Rajakumar, C.</author><author>Ban, M. R.</author><author>Cao, H.</author><author>Young, T. K.</author><author>Bjerregaard, P.</author><author>Hegele, R. A.</author></authors></contributors><auth-address>Vascular Biology Research Group, Robarts Research Institute, University of Western Ontario, London, Ontario, Canada.</auth-address><titles><title>Carnitine palmitoyltransferase IA polymorphism P479L is common in Greenland Inuit and is associated with elevated plasma apolipoprotein A-I</title><secondary-title>J Lipid Res</secondary-title></titles><periodical><full-title>J Lipid Res</full-title></periodical><pages>1223-8</pages><volume>50</volume><number>6</number><edition>2009/02/03</edition><keywords><keyword>Adult</keyword><keyword>Alleles</keyword><keyword>Amino Acid Substitution</keyword><keyword>Apolipoprotein A-I/*blood</keyword><keyword>Atherosclerosis/blood/enzymology/prevention &amp; control</keyword><keyword>Base Sequence</keyword><keyword>Carnitine O-Palmitoyltransferase/blood/deficiency/*genetics</keyword><keyword>Cholesterol, HDL/blood</keyword><keyword>DNA Primers/genetics</keyword><keyword>Female</keyword><keyword>Gene Frequency</keyword><keyword>Genotype</keyword><keyword>Greenland</keyword><keyword>Humans</keyword><keyword>Inuits/*genetics</keyword><keyword>Lipoproteins/blood</keyword><keyword>Male</keyword><keyword>Middle Aged</keyword><keyword>*Mutation, Missense</keyword></keywords><dates><year>2009</year><pub-dates><date>Jun</date></pub-dates></dates><isbn>0022-2275 (Print)&#xD;0022-2275 (Linking)</isbn><accession-num>19181627</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=19181627</url></related-urls></urls><custom2>2681405</custom2><electronic-resource-num>P900001-JLR200 [pii]&#xD;10.1194/jlr.P900001-JLR200</electronic-resource-num><language>eng</language></record></Cite></EndNote>	D<EndNote><Cite><Author>Rajakumar</Author><Year>2009</Year><RecNum>127</RecNum><DisplayText>(15)</DisplayText><record><rec-number>127</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">127</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Rajakumar, C.</author><author>Ban, M. 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K.</author><author>Madsen, L.</author><author>Klementsen, B.</author><author>Vaagenes, H.</author><author>Kryvi, H.</author><author>Froyland, L.</author><author>Hexeberg, S.</author><author>Berge, R. K.</author></authors></contributors><auth-address>Department of Zoology, University of Bergen, Norway. geir.totland@zoo.uib.no</auth-address><titles><title>Proliferation of mitochondria and gene expression of carnitine palmitoyltransferase and fatty acyl-CoA oxidase in rat skeletal muscle, heart and liver by hypolipidemic fatty acids</title><secondary-title>Biol Cell</secondary-title></titles><periodical><full-title>Biol Cell</full-title></periodical><pages>317-29</pages><volume>92</volume><number>5</number><edition>2000/11/09</edition><keywords><keyword>Acyl-CoA Oxidase</keyword><keyword>Animals</keyword><keyword>Carnitine O-Palmitoyltransferase/genetics/*metabolism</keyword><keyword>Diaphragm/cytology/drug effects/enzymology/metabolism</keyword><keyword>Docosahexaenoic Acids/administration &amp; dosage/pharmacology</keyword><keyword>Eicosapentaenoic Acid/administration &amp; dosage/pharmacology</keyword><keyword>Fatty Acids/administration &amp; dosage/*pharmacology</keyword><keyword>Gene Expression Regulation, Enzymologic/*drug effects</keyword><keyword>Hepatocytes/drug effects/enzymology/metabolism</keyword><keyword>Hypolipidemic Agents/administration &amp; dosage/*pharmacology</keyword><keyword>Liver/cytology/drug effects/enzymology/metabolism</keyword><keyword>Male</keyword><keyword>Microscopy, Electron</keyword><keyword>Mitochondria/drug effects/enzymology/genetics/*metabolism</keyword><keyword>Muscle Fibers, Skeletal/cytology/drug effects/enzymology/metabolism</keyword><keyword>Muscle, Skeletal/cytology/drug effects/enzymology/metabolism</keyword><keyword>Myocardium/cytology/enzymology/metabolism</keyword><keyword>Oxidoreductases/genetics/*metabolism</keyword><keyword>Particle Size</keyword><keyword>Peroxisomes/drug effects/enzymology/metabolism/ultrastructure</keyword><keyword>RNA, Messenger/genetics/metabolism</keyword><keyword>Rats</keyword><keyword>Rats, Wistar</keyword><keyword>Sulfides/administration &amp; dosage/pharmacology</keyword></keywords><dates><year>2000</year><pub-dates><date>Aug</date></pub-dates></dates><isbn>0248-4900 (Print)&#xD;0248-4900 (Linking)</isbn><accession-num>11071041</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=11071041</url></related-urls></urls><electronic-resource-num>S0248490000010777 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>�D<EndNote><Cite><Author>Totland</Author><Year>2000</Year><RecNum>152</RecNum><DisplayText>(20)</DisplayText><record><rec-number>152</rec-number><foreign-keys><key app="EN" db-id="tptpv2txvrewdreved4p9z5yevetrr05ez5a">152</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Totland, G. K.</author><author>Madsen, L.</author><author>Klementsen, B.</author><author>Vaagenes, H.</author><author>Kryvi, H.</author><author>Froyland, L.</author><author>Hexeberg, S.</author><author>Berge, R. K.</author></authors></contributors><auth-address>Department of Zoology, University of Bergen, Norway. geir.totland@zoo.uib.no</auth-address><titles><title>Proliferation of mitochondria and gene expression of carnitine palmitoyltransferase and fatty acyl-CoA oxidase in rat skeletal muscle, heart and liver by hypolipidemic fatty acids</title><secondary-title>Biol Cell</secondary-title></titles><periodical><full-title>Biol Cell</full-title></periodical><pages>317-29</pages><volume>92</volume><number>5</number><edition>2000/11/09</edition><keywords><keyword>Acyl-CoA Oxidase</keyword><keyword>Animals</keyword><keyword>Carnitine O-Palmitoyltransferase/genetics/*metabolism</keyword><keyword>Diaphragm/cytology/drug effects/enzymology/metabolism</keyword><keyword>Docosahexaenoic Acids/administration &amp; dosage/pharmacology</keyword><keyword>Eicosapentaenoic Acid/administration &amp; dosage/pharmacology</keyword><keyword>Fatty Acids/administration &amp; dosage/*pharmacology</keyword><keyword>Gene Expression Regulation, Enzymologic/*drug effects</keyword><keyword>Hepatocytes/drug effects/enzymology/metabolism</keyword><keyword>Hypolipidemic Agents/administration &amp; dosage/*pharmacology</keyword><keyword>Liver/cytology/drug effects/enzymology/metabolism</keyword><keyword>Male</keyword><keyword>Microscopy, Electron</keyword><keyword>Mitochondria/drug effects/enzymology/genetics/*metabolism</keyword><keyword>Muscle Fibers, Skeletal/cytology/drug effects/enzymology/metabolism</keyword><keyword>Muscle, Skeletal/cytology/drug effects/enzymology/metabolism</keyword><keyword>Myocardium/cytology/enzymology/metabolism</keyword><keyword>Oxidoreductases/genetics/*metabolism</keyword><keyword>Particle Size</keyword><keyword>Peroxisomes/drug effects/enzymology/metabolism/ultrastructure</keyword><keyword>RNA, Messenger/genetics/metabolism</keyword><keyword>Rats</keyword><keyword>Rats, Wistar</keyword><keyword>Sulfides/administration &amp; dosage/pharmacology</keyword></keywords><dates><year>2000</year><pub-dates><date>Aug</date></pub-dates></dates><isbn>0248-4900 (Print)&#xD;0248-4900 (Linking)</isbn><accession-num>11071041</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=11071041</url></related-urls></urls><electronic-resource-num>S0248490000010777 [pii]</electronic-resource-num><language>eng</language></record></Cite></EndNote>�D<EndNote><Cite><Author>Shen</Author><Year>2006</Year><RecNum>41</RecNum><DisplayText>(19)</DisplayText><record><rec-number>41</rec-number><foreign-keys><key app="EN" db-id="a5tar9rpatdv2gez2rlppr0fd99tvw90zd22">41</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Shen, Y.</author><author>Chow, J.</author><author>Wang, Z.</author><author>Fan, G.</author></authors></contributors><auth-address>Department of Human Genetics, Institute of Stem Cell Biology and Medicine, David Geffen School of Medicine, UCLA, Los Angeles, CA 90095, USA.</auth-address><titles><title>Abnormal CpG island methylation occurs during in vitro differentiation of human embryonic stem cells</title><secondary-title>Hum Mol Genet</secondary-title></titles><periodical><full-title>Hum Mol Genet</full-title></periodical><pages>2623-35</pages><volume>15</volume><number>17</number><edition>2006/07/28</edition><keywords><keyword>Astrocytes/physiology</keyword><keyword>*Cell Differentiation</keyword><keyword>Cell Line</keyword><keyword>*CpG Islands</keyword><keyword>DNA (Cytosine-5-)-Methyltransferase/physiology</keyword><keyword>*DNA Methylation</keyword><keyword>Embryo, Mammalian/*cytology</keyword><keyword>Gene Expression Profiling</keyword><keyword>Gene Silencing</keyword><keyword>Humans</keyword><keyword>Leukocytes/physiology</keyword><keyword>Neoplasms/genetics/metabolism</keyword><keyword>Neurons/physiology</keyword><keyword>Octamer Transcription Factor-3/genetics</keyword><keyword>Oligonucleotide Array Sequence Analysis</keyword><keyword>Promoter Regions, Genetic</keyword><keyword>Stem Cells/*physiology</keyword></keywords><dates><year>2006</year><pub-dates><date>Sep 1</date></pub-dates></dates><isbn>0964-6906 (Print)&#xD;0964-6906 (Linking)</isbn><accession-num>16870691</accession-num><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&amp;db=PubMed&amp;dopt=Citation&amp;list_uids=16870691</url></related-urls></urls><electronic-resource-num>ddl188 [pii]&#xD;10.1093/hmg/ddl188</electronic-resource-num><language>eng</language></record></Cite></EndNote>�D<EndNote><Cite><Author>Shen</Author><Year>2006</Year><RecNum>41</RecNum><DisplayText>(19)</DisplayText><record><rec-number>41</rec-number><foreign-keys><key app="EN" db-id="a5tar9rpatdv2gez2rlppr0fd99tvw90zd22">41</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Shen, Y.</author><author>Chow, J.</author><author>Wang, Z.</author><author>Fan, G.</author></authors></contributors><auth-address>Department of Human Genetics, Institute of Stem Cell Biology and Medicine, David Geffen School of Medicine, UCLA, Los Angeles, CA 90095, USA.</auth-address><titles><title>Abnormal CpG island methylation occurs during in vitro differentiation of human embryonic stem cells</title><secondary-title>Hum Mol Genet</secondary-title></titles><periodical><full-title>Hum Mol Genet</full-title></periodical><pages>2623-35</pages><volume>15</volume><number>17</number><edition>2006/07/28</edition><keywords><keyword>Astrocytes/physiology</keyword><keyword>*Cell Differentiation</keyword><keyword>Cell Line</keyword><keyword>*CpG Islands</keyword><keyword>DNA (Cytosine-5-)-Methyltransferase/physiology</keyword><keyword>*DNA Methylation</keyword><keyword>Embryo, Mammalian/*cytology</keyword><keyword>Gene Expression Profiling</keyword><keyword>Gene 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