SIRT1 negatively regulates the expression of Prl2C3,a senescence-associated protein

SIRT1 is a mammalian homologue of yeast longevity protein Sir2. SIRT1 deacetylates transcription factors, cofactors, and histones in an NAD+-dependent manner, and promotes cell survival, anti-oxidative function, and DNA repair. Although some studies have indicated that SIRT1 is involved in longevity, the function of SIRT1 for preventing aging and senescence is still unclear. In mouse embryonic fibroblasts (MEFs), we found that SIRT1 expression decreased by aging and IRT1 reciprocally regulated the expression level of Prl2C3, one of the prolactin-like peptides. In young MEFs, purified Prl2C3 inhibited the growth and increased the number of senescence-associated β galactosidase-positive cells with enlarged and flattened shapes. Moreover, immunostaining of human skin sections showed the expression of Prl2C3 in the basal cells of the epidermis. These results indicate that SIRT1 negatively regulates a senescence-associated protein rl2C3

Multiple Endocrine Neoplasia Type 1 Producing Growth Hormone- Releasing Factor in an Endocrine Pancreatic Tumor

We report a very rare case of multiple endocrine neoplasia type 1 (MEN 1) where a pituitary tumor presenting with acromegaly was associated with a growth hormone-releasing factor (GRF) producing pancreatic tumor. Twenty-seven months after surgery for pituitary adenoma, a 27-year-old male visited our hospital complaining of epigastralgia and back pain. Radiological examination revealed a 7-cm tumor in the pancreatic tail. Endocrinological studies revealed an abnormal increase in the level of growth hormone (GH). After resection of the pancreatic tumor, the GH level was normalized. Immunohistochemical studies confirmed the existence of GRF in the tumor. A diagnosis of GRF-producing pancreatic tumor was established. Nine months after surgery for the pancreatic tumor, the GH level remained normal and the pituitary gland had decreased in size. We speculate that secondary hyperpituitarism was caused by GRF produced by the endocrine pancreatic tumor in this case

Estimation of Completion of Dormancy in Grapevine Bud Based on Cumulative Temperature

　This study was conducted to investigate the relationship between completion of dormancy of grapevine bud and temperature. Canes of ‘Kyoho’ and ‘Pione’ grapevines (Vitis labrusca×V. vinifera) grown in 7 vineyards with different temperature conditions, in Nagano, Northern Okayama, Yamanashi, Okayama, Okayama University, Fukuoka and Miyazaki, were collected at three different chilling exposures, December, January and February. These were then sent to Okayama University all at the same time. Cuttings with one bud were put into growth chambers kept at 25 or 30°C with 14 hours daylength, and budbreak in each cutting was surveyed at two day intervals for 60 days. Cumulative chilling hours (CCH) of exposure to below 7.2°C in each treatment time was largest in Nagano, followed in order by Northern Okayama, Yamanashi, Okayama, Okayama University, Fukuoka and Miyazaki. The CCH in Nagano was 2.5 to 4.8 times larger than in Miyazaki depending on the treatment time. The later the treatment time and the higher the temperature, the fewer were the number of days to first budbreak (NDFB) after treatment, irrespective of cultivar. A similar trend was observed in the number of days to 60% budbreak. In ‘Kyoho’ the NDFB was short in Nagano, Okayama University and Miyazaki, and longer in Okayama, Yamanashi and Fukuoka. In ‘Pione’ the NDFB was short in Fukuoka and Okayama University, and longer in Yamanashi and Okayama. The result was a weak negative correlation observed between CCH and NDFB in 4 of 7 vineyards. However, there was a strong positive correlation between NDFB and cumulative temperature (CT), a summation of temperature and hours of exposure to above 0°C from November 1 to treatment time and hours of exposure to 25 or 30°C from start of treatment to budbreak in each plot, in 6 vineyards excluding Miyazaki. The importance of estimating the completion of dormancy in grapevine bud based on CT is discussed.ブドウの芽の休眠完了と温度との関係を調査するため，温度条件の異なる７園（中信農試，山梨果試，岡山農試，岡山農試北部支場，岡山大学，福岡農試および宮崎）で栽培されている‘巨峰’と‘ピオーネ’から低温遭遇量の異なる３時期（12月，１月，２月）に結果母枝を採取した．直ちに岡山大学に送り，１芽を持つ挿し穂に調整した後，25または30℃のインキュベーター（14時間日長）に入れ，２日間隔で60日間発芽を調査した．処理開始時の7.2℃以下の遭遇量は中信農試で最も多く，次いで岡山農試北部支場，山梨果試，岡山農試，岡山大学，福岡農試，宮崎の順で，中信農試と宮崎では処理時期により2.5～4.8倍の差があった．発芽所要日数は，両品種とも処理時期が遅いほど，また温度が高いほど少なく，60％発芽所要日数もほぼ同様の傾向であった．‘巨峰’の発芽所用日数は中信農試，岡山大学および宮崎で少ない一方，岡山農試，山梨果試および福岡農試で多く，また‘ピオーネ’では福岡農試と岡山大学で少なく，山梨果試と岡山農試で多かった．７園のうち４園で低温遭遇量と発芽所要日数との間に負の相関がみられたが，相関係数は低かった．一方，11月以降処理開始までの０℃以上の温度の積算値と処理開始から発芽までの25または30℃での積算値を合計した積算温度と発芽所要日数との間には１園を除き極めて高い正の相関が認められた．これらの結果を基に，積算温度によるブドウの休眠完了予測の可能性を考察した

Cellular analysis of SOD1 protein-aggregation propensity and toxicity: a case of ALS with slow progression harboring homozygous SOD1-D92G mutation

Mutations within Superoxide dismutase 1 (SOD1) cause amyotrophic lateral sclerosis (ALS), accounting for approximately 20% of familial cases. The pathological feature is a loss of motor neurons with enhanced formation of intracellular misfolded SOD1. Homozygous SOD1-D90A in familial ALS has been reported to show slow disease progression. Here, we reported a rare case of a slowly progressive ALS patient harboring a novel SOD1 homozygous mutation D92G (homD92G). The neuronal cell line overexpressing SOD1-D92G showed a lower ratio of the insoluble/soluble fraction of SOD1 with fine aggregates of the misfolded SOD1 and lower cellular toxicity than those overexpressing SOD1-G93A, a mutation that generally causes rapid disease progression. Next, we analyzed spinal motor neurons derived from induced pluripotent stem cells (iPSC) of a healthy control subject and ALS patients carrying SOD1-homD92G or heterozygous SOD1-L144FVX mutation. Lower levels of misfolded SOD1 and cell loss were observed in the motor neurons differentiated from patient-derived iPSCs carrying SOD1-homD92G than in those carrying SOD1-L144FVX. Taken together, SOD1-homD92G has a lower propensity to aggregate and induce cellular toxicity than SOD1-G93A or SOD1-L144FVX, and these cellular phenotypes could be associated with the clinical course of slowly progressive ALS

Physiological function of phospholipase D2 in anti-tumor immunity: regulation of CD8+ T lymphocyte proliferation

Two major phospholipase D (PLD) isozymes in mammals, PLD1 and PLD2, hydrolyze the membrane phospholipid phosphatidylcholine to choline and the lipid messenger phosphatidic acid. Although their roles in cancer cells have been well studied, their functions in tumor microenvironment have not yet been clarified. Here, we demonstrate that PLD2 in cytotoxic CD8+ T cells plays a crucial role in anti-tumor immunity by regulating their cell proliferation. We found that growth of tumors formed by subcutaneously transplanted cancer cells is enhanced in Pld2-knockout mice. Interestingly, this phenotype was found to be at least in part attributable to the ablation of Pld2 from bone marrow cells. The number of CD8+ T cells, which induce cancer cell death, significantly decreased in the tumor produced in Pld2-knockout mice. In addition, CD3/CD28-stimulated proliferation of primary cultured splenic CD8+ T cells is markedly suppressed by Pld2 ablation. Finally, CD3/CD28-dependent activation of Erk1/2 and Ras is inhibited in Pld2-deleted CD8+ T cells. Collectively, these results indicate that PLD2 in CD8+ T cells plays a key role in their proliferation through activation of the Ras/Erk signaling pathway, thereby regulating anti-tumor immunity

Inhibition of Hepatitis C Virus Replication and Viral Helicase by Ethyl Acetate Extract of the Marine Feather Star Alloeocomatella polycladia

Hepatitis C virus (HCV) is a causative agent of acute and chronic hepatitis, leading to the development of hepatic cirrhosis and hepatocellular carcinoma. We prepared extracts from 61 marine organisms and screened them by an in vitro fluorescence assay targeting the viral helicase (NS3), which plays an important role in HCV replication, to identify effective candidates for anti-HCV agents. An ethyl acetate-soluble fraction of the feather star Alloeocomatella polycladia exhibited the strongest inhibition of NS3 helicase activity, with an IC50 of 11.7 µg/mL. The extract of A. polycladia inhibited interaction between NS3 and RNA but not ATPase of NS3. Furthermore, the replication of the replicons derived from three HCV strains of genotype 1b in cultured cells was suppressed by the extract with an EC50 value of 23 to 44 µg/mL, which is similar to the IC50 value of the NS3 helicase assay. The extract did not induce interferon or inhibit cell growth. These results suggest that the unknown compound(s) included in A. polycladia can inhibit HCV replication by suppressing the helicase activity of HCV NS3. This study may present a new approach toward the development of a novel therapy for chronic hepatitis C

Periostin as a novel biomarker for postoperative recurrence of chronic rhinosinitis with nasal polyps

We previously reported that chronic rhinosinusitis with nasal polyps (CRSwNP) was subdivided into four chronic rhinosinusitis (CRS) subtypes using the JESREC scoring system. We sought to identify the gene expression profile and biomarkers related with CRSwNP by RNA-sequence. RNA-sequencing was performed to identify differentially expressed genes between nasal polyps (NPs) and inferior turbinate mucosa from 6 patients with CRSwNP, and subsequently, quantitative real-time PCR was performed to verify the results. ELISA was performed to identify possible biomarkers for postoperative recurrence. In the RNA-sequencing results, periostin (POSTN) expression was the highest in NP. We focused on POSTN and investigated the protein level of POSTN by immunohistochemistry and ELISA. POSTN was diffusely expressed in moderate and severe eosinophilic CRS using immunohistochemistry, and its staining pattern was associated with the severity of the phenotype of the CRSwNP (P < 0.05). There was a significant difference between the POSTN high/low groups for postoperative recurrence when the cutoff point was set at 115.5 ng/ml (P = 0.0072). Our data suggests that the protein expression level of POSTN was associated with the severity of CRSwNP, and serum POSTN can be a novel biomarker for postoperative recurrence of CRSwNP

Plaque REgression with Cholesterol absorption Inhibitor or Synthesis inhibitor Evaluated by IntraVascular UltraSound (PRECISE-IVUS Trial): Study protocol for a randomized controlled trial

AbstractBackgroundAlthough the positive association between achieved low-density lipoprotein cholesterol (LDL-C) level and the risk of coronary artery disease (CAD) has been confirmed by randomized studies with statins, many patients remain at high residual risk of events suggesting the necessity of novel pharmacologic strategies. The combination of ezetimibe/statin produces greater reductions in LDL-C compared to statin monotherapy.PurposeThe Plaque REgression with Cholesterol absorption Inhibitor or Synthesis inhibitor Evaluated by IntraVascular UltraSound (PRECISE-IVUS) trial was aimed at evaluating the effects of ezetimibe addition to atorvastatin, compared with atorvastatin monotherapy, on coronary plaque regression and change in lipid profile in patients with CAD.MethodsThe study is a prospective, randomized, controlled, multicenter study. The eligible patients undergoing IVUS-guided percutaneous coronary intervention will be randomly assigned to receive either atorvastatin alone or atorvastatin plus ezetimibe (10mg) daily using a web-based randomization software. The dosage of atorvastatin will be increased by titration within the usual dose range with a treatment goal of lowering LDL-C below 70mg/dL based on consecutive measures of LDL-C at follow-up visits. IVUS will be performed at baseline and 9–12 months follow-up time point at participating cardiovascular centers. The primary endpoint will be the nominal change in percent coronary atheroma volume measured by volumetric IVUS analysis.ConclusionPRECISE-IVUS will assess whether the efficacy of combination of ezetimibe/atorvastatin is noninferior to atorvastatin monotherapy for coronary plaque reduction, and will translate into increased clinical benefit of dual lipid-lowering strategy in a Japanese population