Aberrantly methylated genes in human papillary thyroid cancer and their association with BRAF/RAS mutation

Cancer arises through accumulation of epigenetic and genetic alteration. Aberrant promoter methylation is a common epigenetic mechanism of gene silencing in cancer cells. We here performed genome-wide analysis of DNA methylation of promoter regions by Infinium HumanMethylation27 BeadChip, using 14 clinical papillary thyroid cancer samples and 10 normal thyroid samples. Among the 14 papillary cancer cases, 11 showed frequent aberrant methylation, but the other three cases showed no aberrant methylation at all. Distribution of the hypermethylation among cancer samples was non-random, which implied existence of a subset of preferentially methylated papillary thyroid cancer. Among 25 frequently methylated genes, methylation status of six genes (HIST1H3J, POU4F2, SHOX2, PHKG2, TLX3, HOXA7) was validated quantitatively by pyrosequencing. Epigenetic silencing of these genes in methylated papillary thyroid cancer cell lines was confirmed by gene re-expression following treatment with 5-aza-2′-deoxycytidine and trichostatin A, and detected by real-time RT-PCR. Methylation of these six genes was validated by analysis of additional 20 papillary thyroid cancer and 10 normal samples. Among the 34 cancer samples in total, 26 cancer samples with preferential methylation were significantly associated with mutation of BRAF/RAS oncogene (P = 0.04, Fisher's exact test). Thus, we identified new genes with frequent epigenetic hypermethylation in papillary thyroid cancer, two subsets of either preferentially methylated or hardly methylated papillary thyroid cancer, with a concomitant occurrence of oncogene mutation and gene methylation. These hypermethylated genes may constitute potential biomarkers for papillary thyroid cancer

Myocardial sympathetic denervation prevents chamber-specific alteration of beta-adrenergic transmembrane signaling in rabbits with heart failure

Objectives.The purpose of this study was to assess the effect of myocardial sympathetic denervation on the chamber-specific alteration of beta-adrenergic signaling in left ventricular failure in rabbits.Background.Local abnormalities in sympathetic nerve terminals, including the neuronal reuptake of norepinephrine, are thought to be responsible for the chamber-specific regulation of beta-adrenergic signaling in heart failure.Methods.Sixteen rabbits were given 6-hydroxydopamine, 25 mg/kg body weight intravenously on days 1 and 2 and 50 mg/kg intravenously on days 7 and 8. Another 16 rabbits received vehicle. Aortic regurgitation was induced in eight of the 6-hydroxydopamine—treated and eight of the vehicle-treated rabbits on day 14. Another eight of the 6-hydroxydopamine—treated and eight of the vehicletreated rabbits underwent a sham operation. The hearts were excised for biochemical analysis on day 21.Results.Hemodynamic characteristics on day 21 showed left ventricular failure in both the aortic regurgitation groups. The plasma norepinephrine concentration on day 21 was higher in both the aortic regurgitation groups than in the sham groups. The beta-adrenoceptor densities and isoproterenol plus 5′guanylylimidodiphosphate-, 5′-guanylylimidodiphosphate- and sodium fluoride-stimulated adenylyl cyclase activities were decreased only in the failing left ventricle of the vehicle-pretreated aortic regurgitation group, but in both ventricles of the 6-hydroxydopamine-pretreated aortic regurgitation group. The basal and forskolin-stimulated adenylyl cyclase activities were similar in both the aortic regurgitation groups and in the sham groups.Conclusions.Sympathetic denervation prevented chamberspecific alterations in beta-adrenergic signaling in acute left ventricular failure. Local loss of sympathetic nerve endings, and especially the defective neuronal norepinephrine reuptake, are likely to be responsible for the chamber-specific alteration of the beta-adrenoceptor-G protein-adenylyl cyclase system in heart failure in rabbits

Mutation Analysis of 2009 Pandemic Influenza A(H1N1) Viruses Collected in Japan during the Peak Phase of the Pandemic

BACKGROUND: Pandemic influenza A(H1N1) virus infection quickly circulated worldwide in 2009. In Japan, the first case was reported in May 2009, one month after its outbreak in Mexico. Thereafter, A(H1N1) infection spread widely throughout the country. It is of great importance to profile and understand the situation regarding viral mutations and their circulation in Japan to accumulate a knowledge base and to prepare clinical response platforms before a second pandemic (pdm) wave emerges. METHODOLOGY: A total of 253 swab samples were collected from patients with influenza-like illness in the Osaka, Tokyo, and Chiba areas both in May 2009 and between October 2009 and January 2010. We analyzed partial sequences of the hemagglutinin (HA) and neuraminidase (NA) genes of the 2009 pdm influenza virus in the collected clinical samples. By phylogenetic analysis, we identified major variants of the 2009 pdm influenza virus and critical mutations associated with severe cases, including drug-resistance mutations. RESULTS AND CONCLUSIONS: Our sequence analysis has revealed that both HA-S220T and NA-N248D are major non-synonymous mutations that clearly discriminate the 2009 pdm influenza viruses identified in the very early phase (May 2009) from those found in the peak phase (October 2009 to January 2010) in Japan. By phylogenetic analysis, we found 14 micro-clades within the viruses collected during the peak phase. Among them, 12 were new micro-clades, while two were previously reported. Oseltamivir resistance-related mutations, i.e., NA-H275Y and NA-N295S, were also detected in sporadic cases in Osaka and Tokyo

CD-DST ホウ オ モチイタ ニュウガン INTRINSIC SUBTYPES ニ オケル コウガンザイ カンジュ セイ シケン ノ ケントウ

乳癌の個別化治療が進む中で,依然としてエストロゲンレセプター (ER) やHER2遺伝子の発現状況と化学療法の薬剤感受性との関係には不明な点が多い.今回我々はCollagen gel Droplet embedded culture Drug Sensitivity Test (CD-DST法) を用いた抗癌剤感受性試験を行い,乳癌のsubtypeと薬剤感受性の関係について検討を行った.対象は乳癌患者60例59検体で,5-FUは34検体,epirubicinは35検体,paclitaxelは29検体で抗癌剤感受性の評価が可能であった.5-FU,epirubicin,paclitaxelいずれの薬剤においてもHER2発現状況と抗癌剤感受性との間に有意な相関は認められなかった.ERの発現を認めるホルモン陽性乳癌ではepirubicinに対する薬剤感受性が有意に低下し,5-FU,paclitaxelに対しても感受性が低い傾向が認められた.術前化学療法施行例では抗癌剤感受性試験の評価可能率が低下する事が判明した.CD-DST法は今後の乳癌化学療法を選択する上で,有用となりうる可能性が示唆された.Breast cancer is classified into four subtypes according to estrogen receptor (ER) and human epidermal growth factor receptor 2( HER2) expression. The biological subtypes have a spectrum of clinical, pathologic, and molecular features. Here, we investigated the correlation between the breast cancer subtype and chemosensitivity using the collagen gel droplet embedded culture drug sensitivity test (CD-DST). The response to 5-fluorouracil( 5-FU) was examined in 34 specimens;the response to epirubicin, in 35; and the response to paclitaxel, in 29. HER2 overexpression was not significantly correlated to the chemotherapeutic response to 5-FU, epirubicin, or paclitaxel. ER-positive breast cancer was significantly less sensitive to epirubicin and tended to be refractory to 5-FU and paclitaxel. The evaluation rate of the anti-drug sensitivity test was significantly lower in the group that received neoadjuvant chemotherapy. CD-DST could be useful in choosing anti-cancer drug agents for breast cancer patients

Protective effect of geranylgeranylacetone, an inducer of heat shock protein 70, against drug-induced lung injury/fibrosis in an animal model

<p>Abstract</p> <p>Background</p> <p>To determine whether oral administration of geranylgeranylacetone (GGA), a nontoxic anti-ulcer drug that is an inducer of heat shock protein (HSP) 70, protects against drug-induced lung injury/fibrosis <it>in vivo</it>.</p> <p>Methods</p> <p>We used a bleomycin (BLM)-induced lung fibrosis model in which mice were treated with oral 600 mg/kg of GGA before and after BLM administration. Inflammation and fibrosis were evaluated by histological scoring, hydroxyproline content in the lung and inflammatory cell count, and quantification by ELISA of macrophage inflammatory protein-2 (MIP-2) in bronchoalveolar lavage fluid. Apoptosis was evaluated by the TUNEL method. The induction of HSP70 in the lung was examined with western blot analysis and its localization was determined by immunohistochemistry.</p> <p>Results</p> <p>We confirmed the presence of inflammation and fibrosis in the BLM-induced lung injury model and induction of HSP70 by oral administration of GGA. GGA prevented apoptosis of cellular constituents of lung tissue, such as epithelial cells, most likely related to the <it>de novo </it>induction of HSP70 in the lungs. GGA-treated mice also showed less fibrosis of the lungs, associated with the findings of suppression of both production of MIP-2 and inflammatory cell accumulation in the injured lung, compared with vehicle-treated mice.</p> <p>Conclusion</p> <p>GGA had a protective effect on drug-induced lung injury/fibrosis. Disease-modifying antirheumatic drugs such as methotrexate, which are indispensable for the treatment of rheumatoid arthritis, often cause interstitial lung diseases, an adverse event that currently cannot be prevented. Clinical use of GGA for drug-induced pulmonary fibrosis might be considered in the future.</p

Key Points and Pitfalls in Electrocardiographic Diagnosis of Acute Myocardial Infarction

Abstract: Since electrocardiographic features of acute myocardial infarction vary temporally and spatially among patients, there are many pitfalls in electrocardiographic diagnosis of this condition. In order to avoid overlooking acute myocardial infarction, it is necessary to consider characteristic findings, such as augmentation of Twave amplitude, ST elevation, and the appearance of abnormal Qwaves, in light of the time after onset. Observation of time-course changes allows us to noninvasively understand the presence/absence of recanalization and pathological conditions including infarct extension, infarct expansion, and retention of pericardial effusion, as well as to three-dimensionally diagnose the infarct site from changes in waveform in various leads. The latter findings reflect the anatomy of the artery responsible for the infarction. In addition, it is possible to determine the infarct site and the infarct-related artery from the waveforms of premature beats. In recent years, visualized diagnosis of coronary heart disease has become possible by means of improved techniques including coronary angiography and intravascular ultrasound. However, it seems that the well-established qualitative diagnosis of myocardial infarction by electrocardiography will continue to be important

Efficacy of octreotide against chylothorax following lateral neck dissection for thyroid cancer: A case report

Introduction: Postsurgical chylothorax is a rare complication of cervical dissection for thyroid cancer. We report that octreotide, a synthetic analog of somatostatin, is effective in treating chylothorax after thyroid carcinoma surgery. Presentation of case: The patient was a 48-year-old woman who presented to our institution complaining of a left anterior cervical mass. We diagnosed this as thyroid papillary carcinoma and performed a subtotal excision of the thyroid gland with left cervical lymph node dissection. The patient developed dyspnea, and a chest X-ray revealed bilateral chylothorax on Day 4 post-surgery. Octreotide was administered since bilateral chylothorax was noted. A marked decrease in chyle effusion was noted just 3 days after starting octreotide, and after a total of 9 days of treatment, there were no further signs of chylous effusion. Discussion: Octreotide is effective against postsurgical chylothorax; however, if there are no signs of improvement, we believe surgical treatment should be considered. Conclusion: Octreotide should be administered first to treat postsurgical chylothorax before surgical treatment is considered