• BMB Reports
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• v.30 no.6
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• pp.390-396
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• 1997

A sandwich-type enzyme-linked immunosorbent assay (ELISA) for the quantification of human apolipoprotein A-I (apoA-I) was developed using monoclonal antibodies. For this assay, we used three monoclonal antibodies to trap and detect apo A-I. HDAI16 and HDA15 monoclonal antibodies were used for trapping apoA-I and HDAI8 monoclonal antibody was for detecting apoA-I. These three monoclonal antibodies were produced by immunizing mice with high density lipoprotein (HDL) isolated from human plasma. By immunoblot analysis, these three monoclonal antibodies were specific to apoA-I and showed no cross-reactivities with other plasma proteins. The results of competition assays for epitope cross-reactivity test also verified that these monoclonal antibodies identified separate and distinct epitopes on HDL and apoA-I. Affinity constants of monoclonal antibodies were measured by ELISA. Their association constants ranged from $10^7$ to $10^8$ $M^{-1}$. For this assay, pure apoA-I was isolated by affinity chromatography using monoclonal antibodies. In this sandwich assay, the amount of HRP-labeled HDAI8 bound to apoA-I trapped by HDAI16 and HDAI5 was proportional to apoA-I concentration in the range of 0 to 500ng/ml. ApoA-I concentration in plasma was calculated from the linear regression equation of standard curve. The precision and reliability of the assays are reflected in the low intra-and interassay coefficients of variation that averaged 3.25% and 4.30%, respectively. This assay is sensitive, simple, reproducible, convenient in incubation interval, and does not use radioisotope: thus it can be widely applied in clinical laboratories.

• Animal cells and systems
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• v.7 no.1
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• pp.69-73
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• 2003

Serum lipid and lipoprotein levels are influenced by genetic factors, and exorcise increases the concentrations of cardio-protective parameters such as high-density lipoprotein cholesterol (HDL-cholesterol) and apolipoproteinAI (apoAI) in human serum. In the present study, we tested the effect of adaptation to endurance exercise on the association of a genetic polymorphism (Pst 1 RFLP) in the apoAI gene with these biochemical parameters. The genotype and allele frequencies for the Pst 1 RFLP were not significantly different between the elite athletes and sedentary controls (P>0.05). There were also no significant associations between the Pst 1 RFLP of the apoAI gene and the biochemical parameters in elite athletic group. Thus, our results suggest that the Pst 1 RFLP of the apoAI gene was not significantly associated with the serum apoAI and HDL-cholesterol concentrations as well as athletic performance in Koreans.

• Journal of Veterinary Science
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• v.21 no.3
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• pp.45.1-45.15
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• 2020

Background: Feline mammary carcinoma is the third most common cancer that affects female cats. Objectives: The purpose of this study was to screen differential serum proteins in feline and clarify the relationship between them and the occurrence of feline mammary carcinoma. Methods: Chinese pastoral cats were used as experimental animals. Six serum samples from cats with mammary carcinoma (group T) and six serum samples from healthy cats (group C) were selected. Differential protein analysis was performed using a Label-free technique, while parallel reaction monitoring (PRM) was performed to verify the screened differential proteins. Results: A total of 82 differential proteins were detected between group T and group C, of which 55 proteins were down regulated and 27 proteins were up regulated. Apolipoprotein A-I, Apolipoprotein A-II (ApoA-II), Apolipoprotein B (ApoB), Apolipoprotein C-III (ApoC-III), coagulation factor V, coagulation factor X, C1q, albumen (ALB) were all associated with the occurrence of feline mammary carcinoma. Differential proteins were involved in a total of 40 signaling pathways, among which the metabolic pathways associated with feline mammary carcinoma were the complement and coagulation cascade and cholesterol metabolism. According to the Label-free results, ApoB, ApoC-III, ApoA-II, FN1, an uncharacterized protein, and ALB were selected for PRM target verification. The results were consistent with the trend of the label-free. Conclusions: This experimen is the first to confirm ApoA-II and ApoB maybe new feline mammary carcinoma biomarkers and to analyze their mechanisms in the development of such carcinoma in feline.

• Journal of Nutrition and Health
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• v.30 no.8
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• pp.936-951
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• 1997

As fat consumption has increased in Korea , serum cholesterol concentrations have risen and the prevalence of atherosclerosis increased. The aim of the present study was to investigate the association among apolipoprotein E (Apo E) phenotypes, dietary fat consumption, plasma lipoprotein levels and fatty acid compositions of red blood cells in young healthy women. Seventy-one percent of participants had Apo E3/3, 14.9 percent had ApoE3/2 , 11.8% had Apo E4/3 and 1.2% had Apo E4.2. The subjects daily consumed approximately 1760kcal containing 29 energy % of fat. The ratio of polyunsaturated fat to saturated fat in a diet was 0. 73 . There were no significant differences of nutrient intakes according to Apo E phenotypes. Total cholesterol concentrations of subjects averaged approximately 160mg/dl , but 12 percent of them had over 200mg/dl, which is higher than the range recommended by the National Cholesterol Deucation Program. Notably, most subjects with 210mg/dl had Apo E4 isoforms. Subjects with Apo E4 isoforms had significantly higher total and LDL-cholesterol concentrations than those with Apo E3/2. Also subjects carrying Apo E4 isoforms tended to accumulated more fat in the body. Their BMI , WHR and arm fat area appeared to be how can arm fat area be greater no you mean grater, please check work vsage than subjects with Apo E3 isoforms, but not significantly. Fat intakes slightly influenced serum cholesterol levels. Myristic aicd intakes were positively correlated to serum total and LDL-choelsterol levels. Polyunsaturated fatty acid intakes were negatively correlated to serum total choelsterollevels. The results of 소냐 study suggest that , people with Apo E4 isoforms need to prevent the raising of serum total and LDL -cholesterol concentrations by reducing calorie and fat intakes and by maintaining a normal weight.

• Nutrition Research and Practice
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• v.10 no.1
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• pp.67-73
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• 2016

BACKGROUND/OBJECTIVES: This study was aimed at examining the association between dietary flavanones intake and lipid profiles according to the presence of metabolic syndrome (MetS) in Korean women with type 2 diabetes mellitus (T2DM). SUBJECTS/METHODS: A cross-sectional analysis was performed among 502 female T2DM patients (non-MetS group; n = 129, MetS group; n = 373) who were recruited from the Huh's Diabetes Clinic in Seoul, Korea between 2005 and 2011. The dietary intake was assessed by a validated semi-quantitative food frequency questionnaire (FFQ) and the data was analyzed using the Computer Aided Nutritional Analysis program (CAN-Pro) version 4.0 software. The intake of flavanones was estimated on the basis of the flavonoid database. RESULTS: In the multiple linear regression analysis after adjustment for confounding factors, daily flavanones intake was negatively associated with CVD risk factors such as total cholesterol, LDL-cholesterol, and apoB and apoB/apoA1 ratio only in the MetS group but not in the non-MetS group. Multiple logistic regression analysis revealed that the odds ratio for a higher apoB/apoA1 ratio above the median (${\geq}0.74$) was significantly low in the $4^{th}$ quartile compared to that in the $1^{st}$ quartile of dietary flavanones intake [OR: 0.477, 95% CI: 0.255-0.894, P for trend = 0.0377] in the MetS group. CONCLUSIONS: Dietary flavanones intake was inversely associated with the apoB/apoA1 ratio, suggesting a potential protective effect of flavanones against CVD in T2DM women with MetS.

• Asian-Australasian Journal of Animal Sciences
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• v.9 no.5
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• pp.515-517
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• 1996

The relative efficiencies of deposition into egg yolk of apo-carotenoic acid ester(APO-E, CAROPHYLL Yellow) and saponified marigold xanthophylls (MX), in the presence of canthaxanthin (CN), were determined using a wheat-based diet. APO-E was deposited with an efficiency of 50 percent and MX with an efficiency between 13 and 20 percent. The dose response relationship for MX was curvilinear with a decreased efficiency at higher concentrations. Canthaxanthin was deposited with and efficiency of 38 percent, irrespective of the source of yellow xanthophylls, up to a dietary concentration of 5.5 mg/kg. At a dietary MX concentration of 8.3 mg/kg the efficiency of deposition of CN declined to 24 percent. The results confirm that the replacement ratio of MX : APO-E is between 3 : 1 and 4 : 1 depending on the dietary inclusion of marigold pigment.

• Bulletin of the Korean Chemical Society
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• v.32 no.9
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• pp.3425-3428
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• 2011

Apolipoprotein A-I (Apo A-I) is a major component for high density lipoproteins (HDL). A number of mimetic peptides of Apo A-I were screened from the phase-displayed random peptide library by utilizing monoclonal antibodies (A12). Mimetic peptide for A12 epitope against Apo A-I was selected as CPFARLPVEHHDVVGL (pA1). From the BLAST search, the mimetic peptide pA1 had 40% homology with Apo A-I. As a result of the structural determination of this mimotope using homo/hetero nuclear 2D-NMR techniques and NMR-based distance geometry (DG)/molecular dynamic (MD) computations, DG structure had low penalty value of 0.3-0.7 ${\AA}^2$ and the total RMSD was 0.6-1.6 ${\AA}$. The mimotope pA1 exhibited characteristic conformation including a ${\beta}$-turn from Pro[7] to His[11].

• Journal of Aquaculture
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• v.20 no.1
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• pp.65-72
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• 2007

Full length complementary DNA encoding apolipoprotein A-I (apoA-I) was isolated and characterized in mud loach (Misgurnus mizolepis). Mud loach apoA-I cDNA encoding 24 bp of 5'-untranslated region (UTR), 762 bp of single open reading frame (ORF) consists of 254 amino acids and 293 bp of 3'-UTR excluding stop codon and poly (A+) tail. Two overlapping polyadenylation signals (AATAAAATAAA) was found 9 bp prior to the poly (A+) tail. Mud loach apoA-I represented considerable homology to those from other teleost species at amino acid level with conserving common features of vertebrate apoA-I. Molecular phylogenetic analysis inferred the phylogenetic hypothesis that was generally in accordance with the previous taxonomic relationship. Apolipoprotein A-I mRNA was detected in various tissues, but the mRNA levels were quite varied depending on tissues based on semi-quantitative RT-PCR. Liver and brain showed the significantly higher levels of apoA-I transcripts than other tissues. mRNA expression of apoA-I was quite low in very early stage of embryonic development, however dramatically enhanced from 8 hours post fertilization. This increased mRNA level was retained consistently up to 14 days post hatching.

• Molecules and Cells
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• v.26 no.3
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• pp.291-298
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• 2008

Human karyopherin ${\beta}3$, highly homologous to a yeast protein secretion enhancer (PSE1), has often been reported to be associated with a mediator of a nucleocytoplasmic transport pathway. Previously, we showed that karyopherin ${\beta}3$ complemented the PSE1 and KAP123 double mutant. Our research suggested that karyopherin beta has an evolutionary function similar to that of yeast PSE1 and/or KAP 123. In this study, we performed yeast two-hybrid screening to find a protein which would interact with karyopherin ${\beta}3$ and identified apolipoprotein A-I (apo A-I), a secretion protein with a primary function in cholesterol transport. By using in vitro binding assay, co-immunoprecipitation, and colocalization studies, we defined an interaction between karyopherin ${\beta}3$ and apo A-I. In addition, overexpression of karyopherin ${\beta}3$ significantly increased apo A-I secretion. These results suggest that karyopherin ${\beta}3$ plays a crucial role in apo A-I secretion. These findings may be relevant to the study of a novel function of karyopherin ${\beta}3$ and coronary artery diseases associated with apo A-I.

• BMB Reports
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• v.39 no.5
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• pp.636-641
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• 2006

Our previous studies indicated that native carbonic anhydrase does not interact with hydrophobic adsorbents and that it acquires this ability upon denaturation. In the present study, an apo form of the enzyme was prepared by removal of zinc and a comparative study was performed on some characteristic features of the apo and native forms by far- and near-UV circular dichroism (CD), intrinsic fluorescent spectroscopy, 1-anilino naphthalene-8-sulfonate (ANS) binding, fluorescence quenching by acrylamide, and Tm measurement. Results indicate that protein flexibility is enhanced and the hydrophobic sites become more exposed upon conversion to the apo form. Accordingly, the apo structure showed a greater affinity for interaction with hydrophobic adsorbents as compared with the native structure. As observed for the native enzyme, heat denaturation of the apo form promoted interaction with alkyl residues present on the adsorbents and, by cooling followed by addition of zinc, catalytically-active immobilized preparations were obtained.