The intriguing results described by Petersen et al.3 provide a foundation for further study. Several questions remain to

be answered, however. What factors, genetic or otherwise, allow the development of steatohepatitis and hepatic fibrosis, and what role does IR play in this process? The G allele variant of PNPLA3 has been shown to be associated with the severity of NAFLD but is not associated with IR.8 Conversely, the SNPs in APOC3 have now been linked to NAFLD and IR, but Dabrafenib datasheet so far, there are no data linking APOC3 variations to the severity of NAFLD. A future study combining tests for multiple SNPs linked to NAFLD with hepatic histology is essential to determine the relationship between this and other genetic variations and NAFLD/nonalcoholic steatohepatitis (NASH). Additionally, it is possible that there are specific genetic SNPs that confer protection against hepatic steatosis, steatohepatitis, or both. It can be postulated that in fact this is the case in patients who are phenotypically predisposed to NAFLD but do not have hepatic steatosis. The role of IR in

hepatic steatosis and NAFLD has yet to be fully understood. It is known that patients with NAFLD have increased IR, both systemically and intrahepatically, but it remains uncertain if this is a cause or effect of hepatic steatosis. Petersen et al.3 suggested

that in patients with a normal body mass index, genetic variation in APOC3 leads to hypertriglyceridemia, which in turn causes hepatic steatosis and consequently RO4929097 mw leads to IR. As evidence, they noted that among the 上海皓元医药股份有限公司 members of their small group who lost weight, hepatic steatosis was reduced with a subsequent improvement in IR. This is by no means resounding proof of cause and effect, and recent data from mouse and human studies suggest that IR is independent of hepatic steatosis.9, 10 A complete picture of the interplay between hepatic steatosis and IR remains to be seen, but the reality is likely much more complex and involves not only triglyceride accumulation in the liver and IR but also the host’s defense and repair responses to the potentially hepatotoxic triglyceride precursor molecules (Fig. 1).11 IR not only is a byproduct of hepatic triglyceride excess but also appears to promote hepatic endoplasmic reticulum stress, which may in turn lead to steatohepatitis and even fibrosis.12 In summary, Petersen et al.3 have opened the door to further research aimed at mapping genes associated with NAFLD and, most usefully, advanced disease as evidenced by NASH and fibrosis. This may lead to the ability to predict who is most at risk for progression to cirrhosis or even the development of liver cancer.

The intriguing results described by Petersen et al.3 provide a foundation for further study. Several questions remain to

be answered, however. What factors, genetic or otherwise, allow the development of steatohepatitis and hepatic fibrosis, and what role does IR play in this process? The G allele variant of PNPLA3 has been shown to be associated with the severity of NAFLD but is not associated with IR.8 Conversely, the SNPs in APOC3 have now been linked to NAFLD and IR, but Selleckchem PD0325901 so far, there are no data linking APOC3 variations to the severity of NAFLD. A future study combining tests for multiple SNPs linked to NAFLD with hepatic histology is essential to determine the relationship between this and other genetic variations and NAFLD/nonalcoholic steatohepatitis (NASH). Additionally, it is possible that there are specific genetic SNPs that confer protection against hepatic steatosis, steatohepatitis, or both. It can be postulated that in fact this is the case in patients who are phenotypically predisposed to NAFLD but do not have hepatic steatosis. The role of IR in

hepatic steatosis and NAFLD has yet to be fully understood. It is known that patients with NAFLD have increased IR, both systemically and intrahepatically, but it remains uncertain if this is a cause or effect of hepatic steatosis. Petersen et al.3 suggested

that in patients with a normal body mass index, genetic variation in APOC3 leads to hypertriglyceridemia, which in turn causes hepatic steatosis and consequently find more leads to IR. As evidence, they noted that among the 上海皓元医药股份有限公司 members of their small group who lost weight, hepatic steatosis was reduced with a subsequent improvement in IR. This is by no means resounding proof of cause and effect, and recent data from mouse and human studies suggest that IR is independent of hepatic steatosis.9, 10 A complete picture of the interplay between hepatic steatosis and IR remains to be seen, but the reality is likely much more complex and involves not only triglyceride accumulation in the liver and IR but also the host’s defense and repair responses to the potentially hepatotoxic triglyceride precursor molecules (Fig. 1).11 IR not only is a byproduct of hepatic triglyceride excess but also appears to promote hepatic endoplasmic reticulum stress, which may in turn lead to steatohepatitis and even fibrosis.12 In summary, Petersen et al.3 have opened the door to further research aimed at mapping genes associated with NAFLD and, most usefully, advanced disease as evidenced by NASH and fibrosis. This may lead to the ability to predict who is most at risk for progression to cirrhosis or even the development of liver cancer.

(HEPATOLOGY 2010.) Estrogens promote female reproductive

organ development and function, but estrogen receptors (ERs) are also found, at lower levels, in the skin,1 intestine,2 brain,3 and liver4 where they exert significant influence over diverse aspects of cellular physiology. For example, in the skin, the transition from cyclical estrogen fluctuations during reproductive life to an estrogen deficiency after menopause is associated with dryness, atrophy, fine wrinkling, and poor wound healing.5 JNK inhibitor mouse Estrogens also intimately regulate interleukin-6 (IL-6) expression in various cell types.6 IL-6 is also critical to epithelial barrier function and wound healing in the skin7 and gastrointestinal8 and biliary tracts.9 For example, estrogens inhibit macrophage IL-6 production, which in turn, maintains serum IL-6 levels at relatively low levels during reproductive Gemcitabine purchase years.10 Menopausal loss of estrogens elevates serum IL-6 levels,

which in turn, stimulates osteoclastic bone resorption.6, 11 IL-6−/− mice, however, are resistant to the osteopenic complications of estrogen deficiency.12 In contrast, estrogen stimulates IL-6 production in human ovarian epithelial cells and ovarian cancer cells.13 Promoter complexity controlling IL-6 gene expression14 and complexity of estrogen signaling15, 16 contribute to the tissue-specific regulation of IL-6 expression by estrogens. Estrogens influence biliary tract pathophysiology.17

Females are significantly MCE公司 more susceptible than males to several chronic liver diseases that involve either the biliary tree and/or are influenced by IL-6 expression. Included in these chronic diseases are: (1) primary biliary cirrhosis (PBC),18 a disease associated with variations of IL-6 production19, 20; (2) debilitating/symptomatic adult polycystic liver disease (PCL) requiring liver transplantation,17 in which cyst fluid contains high levels of IL-621; and (3) autoimmune hepatitis, which requires IL-6 production to sustain T helper 17 (TH17)-type T lymphocytes that are critical to disease development.22 These observations raise the hypothesis that estrogens might influence biliary epithelial cell (BEC) IL-6 expression and thereby affect barrier epithelial function, wound repair, and peribiliary or portal tract immune responses. Using primary cultures of non-neoplastic mouse BECs (mBECs) and two human cholangiocarcinoma cell lines, we show that estrogens can stimulate BEC IL-6 production, but only in female or ERα-expressing neoplastic BECs. Estrogen-induced BEC IL-6 production, in turn, is related to ERα expression, which is higher in female than male BECs. Consequently, female BECs are more dependent on the trophic influences of estrogen for continued survival in vitro, and estrogen-induced stimulation of BEC growth can be inhibited by anti–IL-6 antibodies.

2%) and 44 FNBs (51.8%). Diagnosis was achieved similarly between FNA and FNB groups: 75.6% vs. 77.3%, P = 0.86. There were no technical failure cases and procedure related major complications. Final diagnosis of the study patients were

the followings; HKI-272 nmr 72 pancreatic ductal adenocarcinoma (PDAC): 84.7%, 7 neuroendocrine tumor (NET): 8.2%, 4 autoimmune pancreatitis (AIP): 4.7%, 1 metastasis, and 1 chronic pancreatitis: each 1.2%. EUS-FNA and EUS-FNB were used for PDACs (40 FNAs vs. 32 FNBs). Sensitivities were comparable between FNA and FNB groups: 75.0% vs. 81.3%, P = 0.53, and their specificities were 100% in both groups. EUS-FNB was mainly used for AIP or NET to get core biopsy (1 FNA vs. 10 FNBs). Immunohistochemical staining was done for core tissue for pancreas solid lesion suspicious of NET, and they were all compatible with NETs. EUS-FNB provided enough tissue to determine

AIP for a patient. Conclusion: Sensitivity, specificity and safety profiles of FNA and FNB needles were comparable for tissue acquisition of pancreas solid lesion, especially ductal adenocarcinoma. In addition, EUS-FNB might be helpful for diagnosis of NET and AIP. Key Word(s): 1. Pancreas; 2. Mass; 3. EUS-FNA; 4. EUS-FNB; Presenting Author: BIN CHENG www.selleckchem.com/products/AZD6244.html Additional Authors: JINLIN WANG, YAN WANG, RONGHUA WANG Corresponding Author: BIN CHENG Affiliations: Dept. of Gastr., Tongji Hospital of Huazhong University of Science and Technology Objective: EUS-FNA is a safe, sensitive and accurate screening method for mediastinal, retroperitoneal, pancreas and liver lesions. Flow cytometry is a technique that can quantitatively detect the cell surface, intracellular and membrane

antigens, it is a sensitive, rapid and multi-parameter analysis. Limited data exist on the combined use of EUS-FNA and flow cytometry (FC) in the diagnosis of lymphoma. Our aim is to evaluate the performance of EUS- FNA combined with flow cytometry in the diagnosis of mediastinal or retroperitoneal lymphoma. Methods: This study was a retrospective analysis of a collection MCE of data over a 1-year period. Since 2011, 24 patients with lesions suspicious for lymphoma detected by ultrasonography, CT or MRI underwent EUS-FNA and FCM. Results: Of the 24 patients, 22 patients have a clear diagnosis, 8 patients had lymphoma including 7 cases of non-Hodgkin’s lymphoma and 1 case of Hodgkin’s lymphoma, 14 patients had nonlymphoma lesions; for 2 patients, final diagnosis was indeterminate because of insufficient material for FCM. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) accuracy of EUS-FNA combined with flow cytometry for diagnosing lymphoma were respectively 87.5%, 100%, 100%, 93.3%, 95.5%. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) accuracy of EUS-FNA combined with pathology and cytology for diagnosing lymphoma were respectively 25%, 85.7%, 50%, 66.7%, 63.

1C, top). Cre expression was found only in albNScko livers and showed a prominent peak at 2 weeks of age (Fig. 1C, bottom). Histologically, albNScko livers appeared no different from NSflx/flx

livers up to 1 week of age, but began to show increased cellularity around bile ducts at 2 weeks of age (Supporting Fig. 2A-D). When albNScko mice reached 3-4 weeks of age, the liver surface displayed a nodular appearance (Fig. 1D) and showed areas of extensive bile duct hyperplasia (BDH; Fig. 1E1,E2 and Supporting Fig. 2E,F), portal and periportal fibrosis (Supporting Fig. 2G), and necrotic foci in parenchyma (Fig. 1E3). The liver/body weight ratio of albNScko CT99021 cost mice began to exceed that of NSflx/flx mice at 3 weeks (Fig. 1F). These results demonstrate that NS deletion caused liver parenchymal damage and BDH. To establish onset of NS deletion and cell type(s) involved, we examined C59 wnt manufacturer NS expression in

albNScko livers from postnatal day 1 (P1D) to 3 weeks of age. Though a significant number of albNScko hepatocytes still retained their NS expression at P1D, almost all of them lost their NS expression at 1 and 2 weeks of age (Fig. 2A). These findings are consistent with a previous report that differentiated hepatocytes functionally expressing Alb-Cre are rare and distribute in a mosaic pattern in fetal livers.[15] In 2- to 3-week-old albNScko livers, NS-positive cells were mostly confined to the hyperplastic ductular epithelium (Fig. 2B, left panels) and the regenerative hepatic nodules (Fig. 3D1). Contrarily, MCE公司 in the 3-week-old NSflx/flx liver, NS signals were found only in scattered hepatocytes, but not in bile duct epithelial

cells (BECs; Fig. 2B, right panels). Although we cannot exclude the expression of Alb-Cre in subsets of BECs, these results indicate that expression of NS is depleted predominantly in the hepatocytic lineage by albNScko from 1 week of age, but is maintained in the hyperplastic BECs. The time sequence of NSKO-induced events was determined by measuring onset of DNA damage, cell death, and hepatic regeneration in albNScko livers. DNA damage in vivo was detected by the foci formation of phosphorylated histone H2AX (γ-H2AX), which plays a key role in assembling DNA damage response and repair proteins at the damage sites and provides a rapid, sensitive way to detect the DNA damage event.[18] Our results showed that γ-H2AX+ hepatocytes are increased by albNScko as early as 1 week of age (Fig. 3A). This event peaks at 2 weeks and gradually declines afterward, coinciding with the temporal pattern of Cre expression. TUNEL assays showed that the increase of cell death occurs after the DNA damage event and peaks at 3 weeks (Fig. 3B).

1C, top). Cre expression was found only in albNScko livers and showed a prominent peak at 2 weeks of age (Fig. 1C, bottom). Histologically, albNScko livers appeared no different from NSflx/flx

livers up to 1 week of age, but began to show increased cellularity around bile ducts at 2 weeks of age (Supporting Fig. 2A-D). When albNScko mice reached 3-4 weeks of age, the liver surface displayed a nodular appearance (Fig. 1D) and showed areas of extensive bile duct hyperplasia (BDH; Fig. 1E1,E2 and Supporting Fig. 2E,F), portal and periportal fibrosis (Supporting Fig. 2G), and necrotic foci in parenchyma (Fig. 1E3). The liver/body weight ratio of albNScko selleck products mice began to exceed that of NSflx/flx mice at 3 weeks (Fig. 1F). These results demonstrate that NS deletion caused liver parenchymal damage and BDH. To establish onset of NS deletion and cell type(s) involved, we examined selleck kinase inhibitor NS expression in

albNScko livers from postnatal day 1 (P1D) to 3 weeks of age. Though a significant number of albNScko hepatocytes still retained their NS expression at P1D, almost all of them lost their NS expression at 1 and 2 weeks of age (Fig. 2A). These findings are consistent with a previous report that differentiated hepatocytes functionally expressing Alb-Cre are rare and distribute in a mosaic pattern in fetal livers.[15] In 2- to 3-week-old albNScko livers, NS-positive cells were mostly confined to the hyperplastic ductular epithelium (Fig. 2B, left panels) and the regenerative hepatic nodules (Fig. 3D1). Contrarily, MCE in the 3-week-old NSflx/flx liver, NS signals were found only in scattered hepatocytes, but not in bile duct epithelial

cells (BECs; Fig. 2B, right panels). Although we cannot exclude the expression of Alb-Cre in subsets of BECs, these results indicate that expression of NS is depleted predominantly in the hepatocytic lineage by albNScko from 1 week of age, but is maintained in the hyperplastic BECs. The time sequence of NSKO-induced events was determined by measuring onset of DNA damage, cell death, and hepatic regeneration in albNScko livers. DNA damage in vivo was detected by the foci formation of phosphorylated histone H2AX (γ-H2AX), which plays a key role in assembling DNA damage response and repair proteins at the damage sites and provides a rapid, sensitive way to detect the DNA damage event.[18] Our results showed that γ-H2AX+ hepatocytes are increased by albNScko as early as 1 week of age (Fig. 3A). This event peaks at 2 weeks and gradually declines afterward, coinciding with the temporal pattern of Cre expression. TUNEL assays showed that the increase of cell death occurs after the DNA damage event and peaks at 3 weeks (Fig. 3B).

Based on our clinical experience, intramuscular and subcutaneous DHE injections are not as effective as the intravenous route, although, to our knowledge there are no studies that compared the various routes of administration of the drug for CH. The efficacy and tolerability of intranasal DHE (1 mg) in the treatment of acute CH was examined in a controlled study of 25 patients.24 Intranasal DHE decreased the intensity,

but not the duration, of the attacks, and was well tolerated. The authors suggested that the moderate efficacy of the drug in their study may have been related to the dose they used. They recommended that the drug be examined at a higher dose in future trials (the maximal recommended dose of intranasal DHE for acute headache selleck kinase inhibitor treatment in adults is 2 mg). In summary, because of the moderate Cell Cycle inhibitor efficacy of most ergot preparations and the difficulty of receiving intravenous DHE (probably the most effective preparation for this purpose) in a timely manner, the role of ergots in the acute treatment of CH is limited. Data on the efficacy of locally applied lidocaine on acute CH attacks are derived from several non-controlled studies and 1 randomized

controlled trial.25-28 Kittrelle et al examined the effect of lidocaine, applied topically to the sphenopalatine fossa, on acute CH attacks.25 Four of the 5 treated patients experienced rapid relief from pain and associated symptoms of nitrate-induced CH attacks. The treatment was also effective for spontaneous attacks. In another study, Hardebo and Elner examined the effect of lidocaine 4%, self-applied using a nasal dropper through the nostril ipsilateral to the pain, on CH pain and associated symptoms.26 Twenty-four patients were studied, with moderately positive results. Robbins examined the effect of intranasal lidocaine, administered through a spray bottle, on pain in 30 men with ECH.27 Patients treated 2 consecutive CH attacks. Results were modest, with 27% reporting on “moderate MCE公司 relief,” 27% on “mild relief,” and 46% on no relief. In a placebo-controlled study, Costa et al examined the efficacy of lidocaine 10%, applied bilaterally

to the sphenopalatine fossa via a cotton swab using anterior rhinoscopy, on nitroglycerin-induced CH attacks.28 Lidocaine application resulted in elimination of pain in all (15) patients. However, there was a considerable delay (of 37 minutes on average) between the time of lidocaine application and pain relief (the corresponding time interval for placebo was 59 minutes). In summary, intranasal lidocaine is at best moderately effective in the treatment of acute CH attacks. It should not be used as a first-line therapy for this indication. This treatment may be used as adjunctive therapy in some patients whose attacks do not completely respond to other, more effective, therapies. Sicuteri et al conducted a controlled study to examine the efficacy of intravenous somatostatin for acute CH attacks.

Based on our clinical experience, intramuscular and subcutaneous DHE injections are not as effective as the intravenous route, although, to our knowledge there are no studies that compared the various routes of administration of the drug for CH. The efficacy and tolerability of intranasal DHE (1 mg) in the treatment of acute CH was examined in a controlled study of 25 patients.24 Intranasal DHE decreased the intensity,

but not the duration, of the attacks, and was well tolerated. The authors suggested that the moderate efficacy of the drug in their study may have been related to the dose they used. They recommended that the drug be examined at a higher dose in future trials (the maximal recommended dose of intranasal DHE for acute headache Sorafenib treatment in adults is 2 mg). In summary, because of the moderate Navitoclax research buy efficacy of most ergot preparations and the difficulty of receiving intravenous DHE (probably the most effective preparation for this purpose) in a timely manner, the role of ergots in the acute treatment of CH is limited. Data on the efficacy of locally applied lidocaine on acute CH attacks are derived from several non-controlled studies and 1 randomized

controlled trial.25-28 Kittrelle et al examined the effect of lidocaine, applied topically to the sphenopalatine fossa, on acute CH attacks.25 Four of the 5 treated patients experienced rapid relief from pain and associated symptoms of nitrate-induced CH attacks. The treatment was also effective for spontaneous attacks. In another study, Hardebo and Elner examined the effect of lidocaine 4%, self-applied using a nasal dropper through the nostril ipsilateral to the pain, on CH pain and associated symptoms.26 Twenty-four patients were studied, with moderately positive results. Robbins examined the effect of intranasal lidocaine, administered through a spray bottle, on pain in 30 men with ECH.27 Patients treated 2 consecutive CH attacks. Results were modest, with 27% reporting on “moderate 上海皓元医药股份有限公司 relief,” 27% on “mild relief,” and 46% on no relief. In a placebo-controlled study, Costa et al examined the efficacy of lidocaine 10%, applied bilaterally

to the sphenopalatine fossa via a cotton swab using anterior rhinoscopy, on nitroglycerin-induced CH attacks.28 Lidocaine application resulted in elimination of pain in all (15) patients. However, there was a considerable delay (of 37 minutes on average) between the time of lidocaine application and pain relief (the corresponding time interval for placebo was 59 minutes). In summary, intranasal lidocaine is at best moderately effective in the treatment of acute CH attacks. It should not be used as a first-line therapy for this indication. This treatment may be used as adjunctive therapy in some patients whose attacks do not completely respond to other, more effective, therapies. Sicuteri et al conducted a controlled study to examine the efficacy of intravenous somatostatin for acute CH attacks.

Based on our clinical experience, intramuscular and subcutaneous DHE injections are not as effective as the intravenous route, although, to our knowledge there are no studies that compared the various routes of administration of the drug for CH. The efficacy and tolerability of intranasal DHE (1 mg) in the treatment of acute CH was examined in a controlled study of 25 patients.24 Intranasal DHE decreased the intensity,

but not the duration, of the attacks, and was well tolerated. The authors suggested that the moderate efficacy of the drug in their study may have been related to the dose they used. They recommended that the drug be examined at a higher dose in future trials (the maximal recommended dose of intranasal DHE for acute headache selleck compound treatment in adults is 2 mg). In summary, because of the moderate Mdm2 antagonist efficacy of most ergot preparations and the difficulty of receiving intravenous DHE (probably the most effective preparation for this purpose) in a timely manner, the role of ergots in the acute treatment of CH is limited. Data on the efficacy of locally applied lidocaine on acute CH attacks are derived from several non-controlled studies and 1 randomized

controlled trial.25-28 Kittrelle et al examined the effect of lidocaine, applied topically to the sphenopalatine fossa, on acute CH attacks.25 Four of the 5 treated patients experienced rapid relief from pain and associated symptoms of nitrate-induced CH attacks. The treatment was also effective for spontaneous attacks. In another study, Hardebo and Elner examined the effect of lidocaine 4%, self-applied using a nasal dropper through the nostril ipsilateral to the pain, on CH pain and associated symptoms.26 Twenty-four patients were studied, with moderately positive results. Robbins examined the effect of intranasal lidocaine, administered through a spray bottle, on pain in 30 men with ECH.27 Patients treated 2 consecutive CH attacks. Results were modest, with 27% reporting on “moderate MCE公司 relief,” 27% on “mild relief,” and 46% on no relief. In a placebo-controlled study, Costa et al examined the efficacy of lidocaine 10%, applied bilaterally

to the sphenopalatine fossa via a cotton swab using anterior rhinoscopy, on nitroglycerin-induced CH attacks.28 Lidocaine application resulted in elimination of pain in all (15) patients. However, there was a considerable delay (of 37 minutes on average) between the time of lidocaine application and pain relief (the corresponding time interval for placebo was 59 minutes). In summary, intranasal lidocaine is at best moderately effective in the treatment of acute CH attacks. It should not be used as a first-line therapy for this indication. This treatment may be used as adjunctive therapy in some patients whose attacks do not completely respond to other, more effective, therapies. Sicuteri et al conducted a controlled study to examine the efficacy of intravenous somatostatin for acute CH attacks.

Unfortunately, the authors did not Selleck Erlotinib investigate whether recipient rats receiving the serum from injured rats acquired adaptive characteristics of the fibrogenic component of wound healing. They next showed that conditioned media, derived from activated human hepatic stellate cells, have the ability to enhance H2A.Z distribution and H3K27me modifications in Pparg in human mesenchymal stem cells (Fig. 1). This study provides new

evidence that acquired characteristics can be transferred from somatic cells to germ cells through the serum, passing through the Weismann barrier, which strongly supports Lamarckian inheritance. This study raises many questions and further investigation Adriamycin is required. The authors claim that the enrichment of H2A.Z and H3K27me3 at the Pparg locus in sperm from injured male rats were the epigenetic source for adaptation of the hepatic wound-healing response in offspring. If this consideration is true, this epigenetic information ought to spread across all cells that make up individuals in the next generation. The authors monitored the localization of H2A.Z and H3K27me3 at the Pparg locus only in sperm from injured rats and livers (at peak fibrosis) from their injured offspring. To test their hypothesis, epigenetic modifications at the Pparg locus should also be traced from sperm derived from injured rats to early embryos

and various cell types in the offspring. The authors focused on the effect of injured liver only on the adaptation to hepatic disease. However, it is possible 上海皓元 that the serum from injured rats may alter other types of epigenetic information, nonadaptive effects to hepatic disease in their sperm. To test this possibility, genome-wide approaches, such as chromatin immunoprecipitation coupled with massively parallel sequencing (ChIP-Seq), should be used to map the liver injury-induced global changes in epigenetic modification in sperm. Such studies may more comprehensively elucidate epigenetic and nonadaptive effects induced by liver injury. I consider that there are two important

issues in the study of epigenetic transgenerational effects. One is the identification of the “primary epigenetic marks” that are written or erased by environmental cues in germ cells; the other is the extraction of “inheritable epigenetic marks” among primary epigenetic marks beyond a given generation. Epigenetic information is established by the complex crosstalk of transcription factors, epigenetic modifiers, and signal transduction. To identify primary epigenetic marks, it is necessary to exclude secondary effects. Cultured germline stem (GS) cells, which can yield offspring, are a suitable resource with which to identify the primary epigenetic marks established by environmental conditions in germ cells.