FATTY ACID COMPOSITION AND PROSTAGLANDIN CONTENT OF THE RED SEAWEED Gracilaria sp. FROM INDONESIA

High content of polyunsaturated fatty acids (PUFAs) such as arachidonic and eicosapentaenoic acids are typical for the red alga. Analysis of fatty acid composition and prostaglandin content was conducted in the red alga Gracilaria sp. from Indonesia. Total lipid of the alga was extracted with CHCl3-MeOH (2:1, v/v). Analysis of the fatty acids composition was performed on gas chromatography (GC) equipped with omega wax column (30 m x 0,32 mm i.d., Supelco, PA, USA) and analysis of prostaglandins were carried out by HPLC on ODS column (Mightysil RP-18 GP, 250 mm x 4.6 mm, 5 μm). The content of fatty acids high for were palmitic acid (50%) and arachidonic acid (26.9%), whereas prostaglandin E2 was identified and found lower concentration (44.2 μg/gram total lipid)

Identification of DNA methylated regions by using methylated DNA immunoprecipitation sequencing in Brassica rapa

DNA methylation is an epigenetic gene regulatory mechanism that plays an essential role in gene expression, transposon silencing, genome imprinting and plant development. We investigated the influence of DNA methylation on gene expression in Brassica rapa L., to understand whether epigenetic differences exist between inbred lines. Genome-wide DNA methylation was analysed by methylated DNA immunoprecipitation sequencing (MeDIP-seq) of 14-day-old first and second leaves from two inbred lines of Chinese cabbage, one susceptible and one resistant to fusarium yellows caused by Fusarium oxysporum f. sp. conglutinans. MACS (model-based analysis for ChIP-seq) identified DNA methylation peaks in genic regions including 2 kb upstream, exon, intron and 2 kb downstream. More than 65% of genes showed similar patterns of DNA methylation in the genic regions in the two inbred lines. DNA methylation states of the two inbred lines were compared with their transcriptome. Genes having DNA methylation in the intron and in the 200 bp upstream and downstream regions were associated with a lower expression level in both lines. A small number of genes showed a negative correlation between differences in DNA methylation levels and differences in transcriptional levels in the two inbred lines, suggesting that DNA methylation in these genes results in transcriptional suppression

The histone modification H3 lysine 27 tri-methylation has conserved gene regulatory roles in the triplicated genome of Brassica rapa L

© The Author(s) 2019. Published by Oxford University Press on behalf of Kazusa DNA Research Institute. Brassica rapa L. is an important vegetable and oilseed crop. We investigated the distribution of the histone mark tri-methylation of H3K27 (H3K27me3) in B. rapa and its role in the control of gene expression at two stages of development (2-day cotyledons and 14-day leaves) and among paralogs in the triplicated genome. H3K27me3 has a similar distribution in two inbred lines, while there was variation of H3K27me3 sites between tissues. Sites that are specific to 2-day cotyledons have increased transcriptional activity, and low levels of H3K27me3 in the gene body region. In 14-day leaves, levels of H3K27me3 were associated with decreased gene expression. In the triplicated genome, H3K27me3 is associated with paralogs that have tissue-specific expression. Even though B. rapa and Arabidopsis thaliana are not closely related within the Brassicaceae, there is conservation of H3K27me3-marked sites in the two species. Both B. rapa and A. thaliana require vernalization for floral initiation with FLC being the major controlling locus. In all four BrFLC paralogs, low-temperature treatment increases H3K27me3 at the proximal nucleation site reducing BrFLC expression. Following return to normal temperature growth conditions, H3K27me3 spreads along all four BrFLC paralogs providing stable repression of the gene

Atrasentan and renal events in patients with type 2 diabetes and chronic kidney disease (SONAR): a double-blind, randomised, placebo-controlled trial

Background: Short-term treatment for people with type 2 diabetes using a low dose of the selective endothelin A receptor antagonist atrasentan reduces albuminuria without causing significant sodium retention. We report the long-term effects of treatment with atrasentan on major renal outcomes. Methods: We did this double-blind, randomised, placebo-controlled trial at 689 sites in 41 countries. We enrolled adults aged 18–85 years with type 2 diabetes, estimated glomerular filtration rate (eGFR)25–75 mL/min per 1·73 m 2 of body surface area, and a urine albumin-to-creatinine ratio (UACR)of 300–5000 mg/g who had received maximum labelled or tolerated renin–angiotensin system inhibition for at least 4 weeks. Participants were given atrasentan 0·75 mg orally daily during an enrichment period before random group assignment. Those with a UACR decrease of at least 30% with no substantial fluid retention during the enrichment period (responders)were included in the double-blind treatment period. Responders were randomly assigned to receive either atrasentan 0·75 mg orally daily or placebo. All patients and investigators were masked to treatment assignment. The primary endpoint was a composite of doubling of serum creatinine (sustained for ≥30 days)or end-stage kidney disease (eGFR <15 mL/min per 1·73 m 2 sustained for ≥90 days, chronic dialysis for ≥90 days, kidney transplantation, or death from kidney failure)in the intention-to-treat population of all responders. Safety was assessed in all patients who received at least one dose of their assigned study treatment. The study is registered with ClinicalTrials.gov, number NCT01858532. Findings: Between May 17, 2013, and July 13, 2017, 11 087 patients were screened; 5117 entered the enrichment period, and 4711 completed the enrichment period. Of these, 2648 patients were responders and were randomly assigned to the atrasentan group (n=1325)or placebo group (n=1323). Median follow-up was 2·2 years (IQR 1·4–2·9). 79 (6·0%)of 1325 patients in the atrasentan group and 105 (7·9%)of 1323 in the placebo group had a primary composite renal endpoint event (hazard ratio [HR]0·65 [95% CI 0·49–0·88]; p=0·0047). Fluid retention and anaemia adverse events, which have been previously attributed to endothelin receptor antagonists, were more frequent in the atrasentan group than in the placebo group. Hospital admission for heart failure occurred in 47 (3·5%)of 1325 patients in the atrasentan group and 34 (2·6%)of 1323 patients in the placebo group (HR 1·33 [95% CI 0·85–2·07]; p=0·208). 58 (4·4%)patients in the atrasentan group and 52 (3·9%)in the placebo group died (HR 1·09 [95% CI 0·75–1·59]; p=0·65). Interpretation: Atrasentan reduced the risk of renal events in patients with diabetes and chronic kidney disease who were selected to optimise efficacy and safety. These data support a potential role for selective endothelin receptor antagonists in protecting renal function in patients with type 2 diabetes at high risk of developing end-stage kidney disease. Funding: AbbVie

Chiral separation of glycerolipids by high-performance liquid chromatography

Since the enantiomer resolution of synthetic mono- and diacylglycerols as 3,5-dinitrophenylurethanes was achieved by chiral-phase high-performance liquid chromatography (HPLC) in the 1980s, the methodology has been extended to the resolution of various synthetic and naturally occurring chiral glycerolipids. The recent development of polysaccharide-based chiral stationary phases along with optimized chromatographic elution conditions has permitted direct enantiomer resolution of mono-, di-, and triacylglycerols without previous derivatization. However, there are still some compounds for which enantiomer resolution is difficult to achieve, such as phosphatidylcholine and phosphatidylethanolamine. With the development of new derivatization reagents and chromatographic techniques, resolution of these difficult compounds may be possible in the near future. Chiral-phase HPLC will become considerably important in the future as a valuable tool for further expansion of the field of lipidomics and a better understanding of lipid metabolism

脂質の分析II

油脂・脂質の研究では、目的に応じて種々の機器や方法が使用されるが、その中で誰もが一度は経験し、今日広く用いられている便利なものは以下の３つであろう。薄層クロマトグラフィー（TLC)、ガスクロマトグラフィー（GC)、高速液体クロマトグラフィー（HPLC)。何は無くともTLCプレートさえあれば、抽出物にどんな脂質が含まれているか推測できるし、ましてやGCとHPLCが使えたら鬼に金棒、脂肪酸や脂質の精密な分析が可能になる。機器の原理や分析法の詳細は成書・その他に譲るとして、ここではGCとHPLCについて、知っていたら脂質の分析に役立つと思われることの幾つかを（基礎的知見、トラブル対策、ノウハウ等）筆者の経験を交えて紹介する

Recent Advances in Chiral-Phase HPLC for Lipid Analysis

近年、種々の脂質成分について分析法の著しい進歩がみられるが、本稿では脂質の立体異性体（エナンチオマー、ジアステレオマー）分析に大きな役割を果たしているキラル高速液体クロマトグラフィー（気楽固定相を用いるHPLC)の最近の進歩と分析の実際を概説する

Chiral separation of glycerolipids by high-performance liquid chromatography

Since the enantiomer resolution of synthetic mono- and diacylglycerols as 3,5-dinitrophenylurethanes was achieved by chiral-phase high-performance liquid chromatography (HPLC) in the 1980s, the methodology has been extended to the resolution of various synthetic and naturally occurring chiral glycerolipids. The recent development of polysaccharide-based chiral stationary phases along with optimized chromatographic elution conditions has permitted direct enantiomer resolution of mono-, di-, and triacylglycerols without previous derivatization. However, there are still some compounds for which enantiomer resolution is difficult to achieve, such as phosphatidylcholine and phosphatidylethanolamine. With the development of new derivatization reagents and chromatographic techniques, resolution of these difficult compounds may be possible in the near future. Chiral-phase HPLC will become considerably important in the future as a valuable tool for further expansion of the field of lipidomics and a better understanding of lipid metabolism