• Journal of Microbiology and Biotechnology
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• 제32권5호
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• pp.575-581
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• 2022

A Gram-stain-negative, white-coloured, and rod-shaped bacterium, strain DR4-4^T, was isolated from Daechung Reservoir, Republic of Korea, during Microcystis bloom. Strain DR4-4^T was most closely related to Caenimonas terrae SGM1-15^T and Caenimonas koreensis EMB320^T with 98.1% 16S rRNA gene sequence similarities. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between strain DR4-4^T and closely related type strains were below 79.46% and 22.30%, respectively. The genomic DNA G+C content was 67.5%. The major cellular fatty acids (≥10% of the total) were identified as C_16:0, cyclo C_17:0, summed feature 3 (C_16:1ω7c and/or C_16:1ω6c), and summed feature 8 (C_18:1ω7c and/or C_18:1ω6c). Strain DR4-4^T possessed phosphatidylethanolamine, diphosphatidylglycerol, and phosphatidylglycerol as the main polar lipids and Q-8 as the respiratory quinone. The polyamine profile was composed of putrescine, cadaverine, and spermidine. The results of polyphasic characterization indicated that the isolated strain DR4-4^T represents a novel species within the genus Caenimonas, for which the name Caenimonas aquaedulcis sp. nov. is proposed. The type strain is DR4-4^T (=KCTC 82470^T =JCM 34453^T).

• Fisheries and Aquatic Sciences
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• 제25권3호
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• pp.140-150
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• 2022

A method of ultrahigh performance liquid chromatography coupled to high resolution mass spectrometry (UPLC-HRMS) was established for characterization of the lipid profile of Skipjack tuna. Over 300 lipid molecular species were identified through cross-acquisition in both positive and negative ion mode. Phospholipids (PLs) were dominant in Skipjack tuna. Lysophosphatidylethanolamine (LPE), phosphatidylethanolamine (PE), lysophosphatidylcholine (LPC) and phosphatidylcholine (PC) were the main lipid molecular species in PLs, accounting for 89.24% of the total PLs. The ratio of sphingolipids (SLs) and glycerolipids (GLs) were considerable, accounting for 12.30% and 13.60% of the total lipids respectively. Ceramide (Cer) was the main lipid molecular species of SLs, accounting for 64.96% of total SLs, followed by sphingomyelin (SM), accounting for 25.45% of total SLs. Ether diglycerides (ether DG) were the main lipid molecular species of GLs (97.83%). The main fatty acids (FAs) are unsaturated fatty acids (UFAs) in Skipjack tuna. Besides, a new FAs class branched fatty acid esters of hydroxy fatty acids (FAHFA) was detected, together with the FA. The active lipids identified in this study can be used to evaluate the nutritional value of Skipjack tuna.

• Journal of Microbiology and Biotechnology
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• 제32권12호
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• pp.1553-1560
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• 2022

A Gram-stain-negative, rod-shaped bacterial strain, JC4^T, was isolated from a freshwater sample and determined the taxonomic position. Initial identification based on 16S rRNA gene sequences revealed that strain JC4^T is affiliated to the genus Mucilaginibacter with a sequence similarity of 97.97% to Mucilaginibacter rigui WPCB133^T. The average nucleotide identity and digital DNA-DNA hybridization values between strain JC4^T and Mucilaginibacter species were estimated below 80.92% and 23.9%, respectively. Strain JC4^T contained summed feature 3 (C_16:1 ω7c and/or C_16:1 ω6c) and iso-C_15:0 as predominant cellular fatty acids. The dominant polar lipids were identified as phosphatidylethanolamine, one unidentified aminophospholipid, one unidentified phospholipid, and two unidentified lipids. The respiratory quinone was MK-7. The genomic DNA G+C content of strain JC4^T was determined to be 42.44%. The above polyphasic evidences support that strain JC4^T represents a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter aquariorum sp. nov. is proposed. The type strain is JC4^T (= KCTC 92230^T = LMG 32715^T).

• IMMUNE NETWORK
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• 제23권4호
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• pp.28.1-28.20
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• 2023

Lipid accumulation in macrophages is a prominent phenomenon observed in atherosclerosis. Previously, intimal foamy macrophages (FM) showed decreased inflammatory gene expression compared to intimal non-foamy macrophages (NFM). Since reprogramming of lipid metabolism in macrophages affects immunological functions, lipid profiling of intimal macrophages appears to be important for understanding the phenotypic changes of macrophages in atherosclerotic lesions. While lipidomic analysis has been performed in atherosclerotic aortic tissues and cultured macrophages, direct lipid profiling has not been performed in primary aortic macrophages from atherosclerotic aortas. We utilized nanoflow ultrahigh-performance liquid chromatography-tandem mass spectrometry to provide comprehensive lipid profiles of intimal non-foamy and foamy macrophages and adventitial macrophages from Ldlr^-/- mouse aortas. We also analyzed the gene expression of each macrophage type related to lipid metabolism. FM showed increased levels of fatty acids, cholesterol esters, phosphatidylcholine, lysophosphatidylcholine, phosphatidylinositol, and sphingomyelin. However, phosphatidylethanolamine, phosphatidic acid, and ceramide levels were decreased in FM compared to those in NFM. Interestingly, FM showed decreased triacylglycerol (TG) levels. Expressions of lipolysis-related genes including Pnpla2 and Lpl were markedly increased but expressions of Lpin2 and Dgat1 related to TG synthesis were decreased in FM. Analysis of transcriptome and lipidome data revealed differences in the regulation of each lipid metabolic pathway in aortic macrophages. These comprehensive lipidomic data could clarify the phenotypes of macrophages in the atherosclerotic aorta.

• 한국어병학회지
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• 제18권1호
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• pp.67-80
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• 2005

미꾸라지 (mudloach, Misgunus mizolepis)의 간으로부터 클로닝한 phosphoinositide-specific phospholipase C$\delta$ (ML-PLC$\delta$)를 대장균 (E. coli)에서 과발현시켜 만든 재조합 ML-PLC$\delta$와 미꾸라지 간 조직으로부터 직접 정제한 ML-PLC$\delta$의 생화학적 특성을 비교분석하였다. 우선, pET28a vector (Novagen)를 이용하여 E. coli BL21(DE3)에서 과발현된 재조합 ML-PLC$\delta$은 $Ni^{2+}$-NTA affinity 크로마토그래피 및 gel filtration 칼럼에 의해서 정제되었다. 미꾸라지 간 조직으로 ML-PLC$\delta$는 open heparin 칼럼 및 분석용 heparin 칼럼등을 통하여 부분 정제하였다. 두개의 재조합 및 wild ML-PLC$\delta$는 phosphatidylinositol 4,5-bis-phosphate ($PIP_2$)에 대한 농도 의존적 PLC 활성을 보여주었고, 그 활성은 포유류 PLC$\delta$ 효소와 유사하게 칼슘 농도에 의존적인 활성을 나타내었다. 재조합 및 wild ML-PLC$\delta$는 각각 pH 7.0 및 7.5에서 가장 큰 PI-가수분해 활성을 나타낸다는 사실을 알 수 있었다. 게다가, 재조합 및 wild ML-PLC$\delta$는 sodium doecylcholate (SDC) 및 phosphatidylethanolamine (PE), phosphatidylcholine (PC)와 같은 지질류에 대하여 농도의존적인 활성을 나타내나, spermine과 같은 polyamine류의 존재하에서는 농도 의존적으로 PLC 활성이 감소됨을 알 수 있었다. 미꾸라지 각 기관들의 ML-PLC$\delta$의 발현양상 및 양등을 측정하여 보았을 때 ML-PLC$\delta$는 포유류 PLC$\delta$와 마찬가지로 다양한 형태의 PLC$\delta$가 존재함을 알 수 있었다. 이와 같은 결과들로 미루어서 미꾸라지로부터 얻은 ML-PLC$\delta$는 포유류의 PLC$\delta$ isozymes과 유사한 형태의 생화학적 특성을 가지나, 포유류 PLC$\delta$1과 PLC$\delta$3 isozyme의 생화학적 특성을 함께 가짐을 알 수 있었다.

• 생명과학회지
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• 제26권8호
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• pp.875-880
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• 2016

진핵세포의 원형질막은 외부환경으로부터 세포를 격리하는 물리적인 장벽뿐만 아니라, 선택적 물질수송, 신호전달등 중요한 기능을 수행한다. 원형질막의 세포질쪽 지질층은 주로 Phosphatidylethanolamine (PE), Phosphatidylserine (PS), Phosphatidylinositides (PIs) 등의 인지질로 구성되어 있는데, 다양한 단백질들이 이들과 직접 결합하거나 이들의 성질을 이용해 원형질막으로 위치된다. 본 연구에서는 원형질막의 안쪽 지질층에 존재하는 특정 인지질과 결합하여, 원형질막에 위치되는 단백질을 과발현시켰을 때 나타나는 세포의 모양변화를 분석하였다. 이를 위해 PS와 PI등과의 선택적 결합으로 원형질막에 위치하는 단백질들과 소수성 또는 정전기적 상호작용으로 원형질막으로 위치되는 단백질들을 HEK293T세포에 과발현시켜 보았다. 그 결과, 대조군으로 사용한 EGFP 단백질과 PI4P에 선택적으로 결합하는 EGFP-P4M-SidM단백질의 발현은 세포의 모양변화를 유도하지 않았다. 반면, PI(4,5)P₂에 결합하는 EGFP-PLCδ1(PH), PI(3,4,5)P₃에 결합하는 EGFP-AKT1(PH), PI4P와 PI(4,5)P₂에 결합하는 OSH2(PH)x2-EGFP의 발현은 세포의 크기가 줄어드는 수축현상이 일어나면서, Lamilapodia나 Filopodia가 형성되는 것을 확인할 수 있었다. 반면에, PS결합을 통해 원형질막에 위치되는 Lact-C2-EGFP과 소수성결합에 의해 원형질막에 위치되는 ApPDE4 long-form인 L(N20)-EGFP이나, 정전기적인 결합을 통해 원형질막으로 위치되는 ApPDE4 short-form인 S(N-UCR1-2)-EGFP단백질의 경우는 세포의 크기가 줄어드는 수축현상은 일어나지만 Lamilapodia나 Filopodia와 같은 모양변화는 나타나지 않는 것을 확인할 수 있었다. 본 연구를 통해, 특정 인지질에 결합하여 원형질막에 위치되는 단백질들이 각각의 인지질 결합 특성에 따라 분류 가능한 세포의 모양변화가 일어남을 확인할 수 있었다.

• Preventive Nutrition and Food Science
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• 제7권3호
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• pp.265-272
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• 2002

In this study, we examined the effects of dietary fatty acids on the fatty acid composition of phospholipid fractions in regions of the brain and on behavioral development in rats. The Sprague Dawley rats were fed the experimental diets 3~4 wks prior to the conception. Experimental diets consisted of 10% fat(wt/wt) which were from either safflower oil (SO, poor in $\omega$3 fatty acids), mixed oil MO, P/M/S ratio : 1:1.4:1, $\omega$6/$\omega$3 ratio = 6.3), or mixed oil supplemented with vitamin E (+500 mg/kg diet). At 3 and 9 weeks of age, frontal cortex (FC), corpus striatum (CS), hippocampus (H), and cerebellum (CB) were dissected from the whole brain. The fatty acid content was determined in the different phospholipid fractions: phosphatidylcholine (PC), phosphatidyl-serine (PS), and phosphatidylethanolamine (PE) in the rat brain regions. In the visual discrimination test, the order of the cumulative errors made in Y-water maze test were SO > MO > ME. This suggested that the balanced diet supplemented with vitamin I had the most beneficial effect on learning ability. The overall characteristics of correlation between fatty acids and behavior development were that the frequency of cumulative errors were negatively correlated significantly with monounsaturated fatty acids (MUFAs), ie., 18:1 $\omega$9 and 22:1 $\omega$9. Docosa-hexaenoic acid (22:6 $\omega$3) of PS in frontal cortex (FC) was negatively correlated with the number of errors made in the Y-water maze test.22:5 $\omega$6 PS in hippocampus (H), PC and PE in corpus striatum (CS), PC in cerebellum (CB) were positively correlated with cumulative errors. And these errors were negatively correlated with 20:4 $\omega$ 6 of PE in corpus striatum (CS) and PC in cerebellum (CB). Especially, O1eic acid (18:1 u 9) in all phospholipid fractions (PC, PS, PE) of hippocampus was negatively correlated with the number of errors. These findings demonstrate that the MUFAs were might be essential for proper brain development, especially in hippocampus which is generally thought to be the regions of memory and learning.

• Asian-Australasian Journal of Animal Sciences
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• 제31권5호
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• pp.686-695
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• 2018

Objective: The objective of this study was to investigate the effects of dietary choline supplementation on hepatic lipid metabolism and gene expression in finishing pigs with intrauterine growth retardation (IUGR). Methods: Using a $2{\times}2$ factorial design, eight normal birth weight (NBW) and eight IUGR weaned pigs were fed either a basal diet (NBW pigs fed a basal diet, NC; IUGR pigs fed a basal diet, IC) or a diet supplemented with two times more choline than the basal diet (NBW pigs fed a high-choline diet, NH; IUGR pigs fed a high-choline diet, IH) until 200 d of age. Results: The results showed that the IUGR pigs had reduced body weight compared with the NBW pigs (p<0.05 from birth to d 120; p = 0.07 from d 120 to 200). Increased (p<0.05) free fatty acid (FFA) and triglyceride levels were observed in the IUGR pigs compared with the NBW pigs. Choline supplementation decreased (p<0.05) the levels of FFAs and triglycerides in the serum of the pigs. The activities of malate dehydrogenase and glucose 6-phosphate dehydrogenase were both increased (p<0.05) in the livers of the IUGR pigs. Choline supplementation decreased (p<0.05) malate dehydrogenase activity in the liver of the pigs. Gene expression of fatty acid synthase (FAS) was higher (p<0.05) in the IC group than in the other groups, and choline supplementation decreased (p<0.05) FAS and acetyl-CoA carboxylase ${\alpha}$ expression in the livers of the IUGR pigs. The expression of carnitine palmitoyl transferase 1A (CPT1A) was lower (p<0.05) in the IC group than in the other groups, and choline supplementation increased (p<0.05) the expression of CPT1A in the liver of the IUGR pigs and decreased (p<0.01) the expression of hormone-sensitive lipase in both types of pigs. The gene expression of phosphatidylethanolamine N-methyltransferase (PEMT) was higher (p<0.05) in the IC group than in the other groups, and choline supplementation significantly reduced (p<0.05) PEMT expression in the liver of the IUGR pigs. Conclusion: In conclusion, the lipid metabolism was abnormal in IUGR pigs, but the IUGR pigs consuming twice the normal level of choline had improved circulating lipid parameters, which could be related to the decreased activity of nicotinamide adenine dinucleotide phosphate-generating enzymes or the altered expressions of lipid metabolism-related genes.

• Journal of Applied Biological Chemistry
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• 제59권3호
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• pp.221-225
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• 2016

Tolaasin은 Pseudomonas tolaasii에 의해 생성되어 pore를 형성하는 펩티드 독소이며, 인공재배 버섯의 막 구조를 파괴하여 갈반병을 일으킨다. Tolaasin이 막에서 pore를 형성하는 기작이나 특성은 자세히 알려지지 않았으나, 인공 지질이중막에서 tolaasin에 의한 pore 형성은 제시되었다. Tolaasin에 의한 지질막에서의 pore 형성은 드물게 나타났고, 형성된 pore는 불안정하기에 tolaasin pore의 길이와 지질막의 두께가 서로 일치하지 않을 수 있음이 제안되었다. 그러므로, 탄소수가 다른 지방산으로 이루어진 인지질들을 첨가하여 tolaasin에 의한 용혈활성 변화를 측정하였다. 두 개의 decanoic acids (C10:0, 1,2-didecanoyl-sn-glycero-3-phosphoethanolamine; DDPE)와 myristic acids (C14:0, 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), stearic acids(C18:0, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine)로 이루어진 phosphatidylethanolamine들을 적혈구와 tolaasin 펩티드가 포함된 반응용액에 가했을 때, DDPE만이 tolaasin에 의한 용혈활성을 촉진하였으며, 나머지 두 인지질은 효과를 보이지 않았다. DDPE를 다양한 농도로 처리하였을 때, tolaasin에 의한 용혈활성은 농도의존적으로 증가하였다. 중간길이의 지방산으로 구성된 인지질은 tolaasin pore 구조와 막지질 사이에 결합하여 pore 주변의 막을 얇게 함으로써 tolaasin pore를 안정화시킬 것으로 여겨진다. 본 연구의 결과는 중간 크기의 지방산으로 구성된 인지질이 tolaasin pore를 막 구조에서 안정화시킴으로써 활성을 증가시키며, 이것은 적혈구 막에서 tolaasin pore의 길이가 막의 두께보다 조금 짧을 것이라는 사실을 제안한다.

• Biomolecules & Therapeutics
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• 제22권2호
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• pp.129-135
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• 2014

Previously, we reported that lysophosphatidylethanolamine (LPE), a lyso-type metabolite of phosphatidylethanolamine, can increase intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) via type 1 lysophosphatidic acid (LPA) receptor ($LPA_1$) and CD97, an adhesion G-protein-coupled receptor (GPCR), in MDA-MB-231 breast cancer cells. Furthermore, LPE signaling was suggested as like $LPA_1/CD97-G_{i/o}$ proteins-phospholipase $C-IP_3-Ca^{2+}$ increase in these cells. In the present study, we further investigated actions of LPE not only in the $[Ca^{2+}]_i$ increasing effect but also in cell proliferation and migration in MDA-MB-231 breast cancer cells. We utilized chemically different LPEs and a specific inhibitor of $LPA_1$, AM-095 in comparison with responses in SK-OV3 ovarian cancer cells. It was found that LPE-induced $Ca^{2+}$ response in MDA-MB-231 cells was evoked in a different manner to that in SK-OV3 cells in terms of structural requirements. AM-095 inhibited LPE-induced $Ca^{2+}$ response and cell proliferation in MDA-MB-231 cells, but not in SK-OV3 cells, supporting $LPA_1$ involvement only in MDA-MB-231 cells. LPA had significant effects on cell proliferation and migration in MDA-MB-231 cells, whereas LPE had less or no significant effect. However, LPE modulations of MAPKs (ERK1/2, JNK and p38 MAPK) was not different to those by LPA in the cells. These data support the involvement of LPA1 in LPE-induced $Ca^{2+}$ response and cell proliferation in breast MDA-MB-231 cells but unknown GPCRs (not $LPA_1$) in LPE-induced responses in SK-OV3 cells. Furthermore, although LPE and LPA utilized $LPA_1$, LPA utilized more signaling cascades than LPE, resulting in stronger responses by LPA in proliferation and migration than LPE in MDA-MB-231 cells.