Agung Purnomoadi

Comparative Proteomics Analysis Reveals L-Arginine Activates Ethanol Degradation Pathways in HepG2 Cells.

L-Arginine (Arg) is a versatile amino acid that plays crucial roles in a wide range of physiological and pathological processes. In this study, to investigate the alteration induced by Arg supplementation in proteome scale, isobaric tags for relative and absolute quantification (iTRAQ) based proteomic approach was employed to comparatively characterize the differentially expressed proteins between Arg deprivation (Ctrl) and Arg supplementation (+Arg) treated human liver hepatocellular carcinoma (HepG2) cells. A total of 21 proteins were identified as differentially expressed proteins and these 21 proteins were all up-regulated by Arg supplementation. Six amino acid metabolism-related proteins, mostly metabolic enzymes, showed differential expressions. Intriguingly, Ingenuity Pathway Analysis (IPA) based pathway analysis suggested that the three ethanol degradation pathways were significantly altered between Ctrl and +Arg. Western blotting and enzymatic activity assays validated that the key enzymes ADH1C, ALDH1A1, and ALDH2, which are mainly involved in ethanol degradation pathways, were highly differentially expressed, and activated between Ctrl and +Arg in HepG2 cells. Furthermore, 10 mM Arg significantly attenuated the cytotoxicity induced by 100 mM ethanol treatment (P < 0.0001). This study is the first time to reveal that Arg activates ethanol degradation pathways in HepG2 cells.

Quantitative proteomics analysis reveals glutamine deprivation activates fatty acid β-oxidation pathway in HepG2 cells

Glutamine, a multifunctional amino acid, functions in nutrient metabolism, energy balance, apoptosis, and cell proliferation. Lipid is an important nutrient and controls a broad range of physiological processes. Previous studies have demonstrated that glutamine can affect lipolysis and lipogenesis, but the effect of glutamine on the detailed lipid metabolism remains incompletely understood. Here, we applied the quantitative proteomics approach to estimate the relative abundance of proteins in HepG2 cells treated by glutamine deprivation. The results showed that there were 212 differentially abundant proteins in response to glutamine deprivation, including 150 significantly increased proteins and 62 significantly decreased proteins. Interestingly, functional classification showed that 43 differentially abundant proteins were related to lipid metabolism. Further bioinformatics analysis and western blotting validation revealed that lipid accumulation may be affected by β-oxidation of fatty acid induced by glutamine deprivation in HepG2 cells. Together, our results may provide the potential for regulating lipid metabolism by glutamine in animal production and human nutrition. The MS data have been deposited to the ProteomeXchange Consortium with identifier PXD003387.

Reduction of methane production from ongole crossbreed cattle in Indonesia by increasing the concentrate feeding frequency

Publisher Summary More than half the livestock population in the world is raised in the tropics. They ingest a low quality of feed due to the high lignifications of forages. Therefore, livestock in the tropics has been pointed to produce higher methane per unit of dry matter intake. Optimizing the reduction of methane conversion ratio (MCR) in the tropics by feeding management, such as increased feeding frequency, has been suggested to improve rumen microbial synthesis. This chapter focuses on a study in which Ongole Crossbreed young bulls were used to study the effect of feeding frequency of concentrate on the reduction of methane. The bulls in the study were allowed a diet of Napier grass (Pennisetum purpureum sp) and concentrate feed at a 40:60 ratio to meet the dry matter (DM) intake of 3.5% of liveweight. The cattle were separated into two groups: one given the concentrate once a day and the other given the concentrate twice a day. The grass to both groups was given twice a day. The parameters of collected data were daily intake of dry matter (DMI), gross energy intake (GEI), methane production, and liveweight gain. These data showed that the feeding management, such as increasing feeding frequency of concentrate, could reduce methane production (MCR), provide better feed utilization (FCR), and increase the animal productivity (liveweight gain). Thus, the study suggests that the method could be adopted to mitigate methane production in the tropics.

Effects of Proportion of Forage in the Diet on the Dry Matter Intake of Holstein Dry Cows During Last 9 Weeks of Pregnancy

経産ホルスタイン種乾乳牛8頭(体重:672.1±95.1kg,産次:4.1±1.7産)を用いて,分娩予定13週前から分娩までイタリアンライグラス1番草の乾草ウエハーと配合飼料の比率が9:1(H区,n=3,1頭除外)または5:5(C区,n=4)となるように混合して給与し,乾物摂取量(DMI)を測定した.飼料は,可消化養分総量(TDN)でホルスタイン種の維持要求量にホルスタイン種の胎子1頭増給分を満たすものとした.H区では分娩4週前以降に残飼が観察され,分娩が近づくに従ってその量が大きく増加し,乾物給与量に対するDMIの割合は分娩1週前に低くなる傾向(P<0.10)がみられた.C区では,全頭が給与飼料を全量採食した.仮に,本試験のH区におけるDMIを,分娩前各週における限界乾物摂取量とすると,分娩前4週間における乾乳牛のエネルギー要求量を満たす飼料中TDN含量は63.1%と計算される.

Effects of Energy Level on Plasma Hormones and Metabolites During the Last Two Months of Pregnancy in Holstein Dairy Cows

ホルスタイン種経産妊娠牛26頭を用いて,乳牛の妊娠末期におけるエネルギー摂取量が血中代謝産物およびホルモン濃度に及ぼす影響について検討した.イタリアンライグラス2番乾草またはサイレージ,および配合飼料を可消化養分総量(TDN)でホルスタイン種母体の維持要求量(M区),または維持要求量に胎子1頭分を増給(MP区)するものとした飼養試験を,妊娠28週目から行った.妊娠30週から40週まで2週に1度頸静脈および尾動脈から,それぞれ採食前に採血し,代謝産物濃度およびホルモン濃度の変化について検討した.M区の血漿インスリン濃度は,妊娠32週以降はMP区より有意に低く,妊娠36週以降は妊娠の進行に従って低下する傾向を示した.血漿グルコース濃度は,有意な差は観察されなかった.M区の遊離脂肪酸(FFA)濃度は,試験期間中は常にMP区より高い傾向にあり,妊娠40週目に上昇した.MP区では,妊娠38週まではほとんど変化はみられなかったが,分娩直前の40週に急激に上昇した.妊娠34週目以降におけるM区の各妊娠ステージでは,TDN摂取量が低くなるに従って,インスリン濃度は低く,グルカゴン濃度は高く,インスリンとグルカゴンのモル比率は低くなり,グルカゴン作用が優勢となっていた.以上の結果から,妊娠に必要なエネルギーを増給せず,維持量のみで妊娠末期を飼養した乳牛ではインスリンおよびグルカゴン濃度が低く,FFA濃度が高いことから体脂肪の動員が確認された.