• Title/Summary/Keyword: Labelling condition

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Protein Carboxyl Ο-Methylation in Porcine Liver and Testis (돼지 간 및 정소에서 단백질 카르복실메칠화 현상)

  • 조재열;김성수;이향우;홍성렬
    • YAKHAK HOEJI
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    • v.45 no.1
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    • pp.46-54
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    • 2001
  • Protein carboxyl Ο-methylation is a kind of enzymatic reaction producing carboxyl methylester catalyzed by protein carboxyl Ο-methyltransferases at the carboxyl group of amino acid residues in polypeptide. Since the finding of carboxyl methylesterl many studies have been focused on the under-standing of biological functions in eukaryotes but still not clear except for roles in Ras attachment to membrane and protein repair. In this study, we investigated the protein carboxyl methylation in porcine liver and testis in respect of identification and characterization of carboxyl methylesters and natural proteinous substrates using pH stability of the esters and electrophoresis under acidic and basic conditions. We detected several kinds of methyl esters, 3 kinds each in cytosolic fractions from liver and testis. Under the treatment of strong acid and base, the ratio between base-stable substrates and unstable ones in liver (4 : 6) was different from the ratio obtained in testis (6 : 4). The methyl accepting capacities were affected by enzymatic proteolysis between the range of 55 to 65% in liver and of 35 to 45% in testis. Separation of the methylated proteins by acidic electrophoresis in the presence of urea and SDS revealed distinctively natural substrates of 26, 33 and 80 kD in the cytosol from liver and of 14, 25, 32 and 86 kD from testis. Most of the labelling, however were lost following electrophoresis under moderate alkaline condition, except for molecules of newly detected 7 and 17 kD in livers and 15, 29, 40 and 80 kD in testis. From these results, it was proposed that protein carboxyl Ο-methylation in each organs may be catalyzed by different classes of protein carboxyl Ο-methyltransferases. In addition, it is suggested that the protein carboxyl methylation in liver and testis may have different patterns in respect of natural substrates.

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Can Dining Alone Lead to Healthier Menu Item Decisions than Dining with Others? The Roles of Consumption Orientation and Menu Nutrition Information (혼밥이 건강한 메뉴 선택에 미치는 영향: 소비 목적 지향과 메뉴 영양 정보 표시의 역할)

  • Her, EunSol;Behnke, Carl;Almanza, Barbara
    • Korean Journal of Community Nutrition
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    • v.26 no.3
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    • pp.155-166
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    • 2021
  • Objectives: Driven by a growth of single-person households and individualized lifestyles, solo dining in restaurants is an increasingly recognizable trend. However, a research gap exists in the comparison of solo and group diners' menu-decision making processes. Based on the self-control dilemma and the temporal construal theory as a theoretical framework, this study compared the ordering intentions of solo vs. group diners with healthy vs. indulgent (less healthy) entrées. The mediating role of consumption orientation and the moderating role of amount of menu nutrition information were further explored to understand the mechanism and a boundary condition. Methods: A scenario-based online survey was developed using a 2 (dining social context: solo vs. with others) × 3 (amount of menu nutrition information: no nutrition information vs. calories vs. calories/fat/sodium), between-subjects, experimental design. Consumers' level of nutrition involvement was controlled. A nationwide survey data (n = 224) were collected from a crowdsourcing platform in the U.S. Data were analyzed using multivariate analysis of covariance, independent t-test, univariate analysis of covariance, and moderated mediation analyses. Results: Findings reveal that solo (vs. group) diners have less (vs. more) intentions to order indulgent menu items due to a more utilitarian (vs. more hedonic) consumption orientation in restaurant dining. Findings also show that solo (vs. group) diners have more (vs. less) intentions to order healthy menu items when the restaurant menu presented nutrition information including calories, fat, and sodium. Conclusions: The findings contribute to the literature of foodservice management, healthy eating, and consumer behavior by revealing a mechanism and an external stimuli of solo vs. group diners' healthy menu-decision making process in restaurants. Furthermore, the findings provide restauranteurs and health professionals with insights into the positive and negative impacts of menu nutrition labelling on consumers' menu-decisions.

Cryopreservation of Mouse IVF Zygotes by Vitrification (체외수정된 생쥐 1-세포기 배의 초자화 동결)

  • 김묘경;이현숙;엄상준;김은영;윤산현;박세필;정길생;임진호
    • Korean Journal of Animal Reproduction
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    • v.20 no.2
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    • pp.119-126
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    • 1996
  • This study was carried out to determine the optimal condition for successful and efficient c cryopreservation of zygotes, 1-cell embryos, using EFS40 which was 40% (v/v) ethylene glycol diluted in DPBS medium containing 30% Fic-oll (w/v) and 0.3 M sucrose. After mouse zygote produced by IVF was vitrified by two freezing methods, the post-warming survival rates of 1-cell zygotes were assessed as cleavage to the 2-cell stage and development into the hatching blastocysts at 5 day. In the one-step method, when embryos were directly exposed to the vitrification solution at 25$^{\circ}C$ for 1 min., survival and development rates of zygotes were 85.5% and 31.9% In the two-step method, embryos were equilibrated with a dilute 20% EG for 1, 3, 5 min. before 1 min. exposure to EFS40, re-spectively. However, the rates of development (17.7, 3.3, 0%) were lower than that of one-step method. The highest survival rate (95.9%) was obtained by one-step method which exposes embryos in EFS40 for 30 sec. In this condition, 63. 8% of cleaved 2-cell developed into hatching blastocysts. In the cell number of Total and ICM using differential labelling technique, there are no significant differences in the cell number of Total and ICM between blastocysts devel oped in vitrified-thawed embryos (63.2${\pm}$16.9, 1 13.5${\pm}$4.0) and control balstocysts (54.0${\pm}$15.2, 1 12.3${\pm}$4.6). Therefore, these results show that mouse zygotes can be successfully cryopreserved by a simple vitrification method although developmental rates of vitrified embryos were reduced. In conclusion, this proposed vitrifi cation procedures can be useful in the cryopreservation of mouse IVF zygotes.

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Preparation of $^{99m}Tc-HYNIC-PEG-liposomes$ for Imaging of the Focal Sites of Infection (농양 진단을 위한 $^{99m}Tc-HYNIC-PEG-liposomes$의 제조)

  • Hong, Jun-Pyo;Awh, Ok-Doo;Kim, Hyun-Suk;Lee, Eun-Sook;Lee, Tae-Sup;Choi, Tae-Hyun;Choi, Chang-Woon;Lim, Sang-Moo
    • The Korean Journal of Nuclear Medicine
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    • v.36 no.6
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    • pp.333-343
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    • 2002
  • Purpose: A new linker, hydrazino nicotinamide (HYNIC), was recently introduced for labelling of liposome with $^{99m}Tc$. In this study we synthesized HYNIC derivatized PEG (polyethylene glycol)-liposomes radiolabeled with $^{99m}Tc$. Materials and Methods: In order to synthesize HYNIC-DSPE (distearoyl phosphatidyl ethanolamine) which is a crucial component for $^{99m}Tc$ chelation, first of all succinimidyl 6-BOC-hydrazinopyridine-3-carboxylic acid was synthesized from 6-chloronicotinic acid by three sequential reactions. A DSPE derivative of succinimidyl 6-BOC-hydrazinopyridine-3-carboxylic acid was transformed into HYNIC-DSPE by HCI/dioxane. HYNIC-PEG-liposomes were prepared by hydration of the dried lipid mixture of EPC (egg phosphatidyl choline): PEG-DSPE : HYNIC-DSPE:cholesterol (1.85:0.15:0.07:1, molar ratio). The HYNIC-PEG-liposomes were labeled with $^{99m}Tc$ in the presence of $SnCl_2{\cdot}2H_2O$ (a reducing agent) and tricine (a coligand). To investigate the level of in vivo transchelation of $^{99m}Tc$ in the liposomes, the $^{99m}Tc$-HYNiC-PES-liposomes were incubated with a molar excess of DTPA, cysteine or glutathione solutions at $37^{\circ}C$ for 1 hour. The radiolabeled liposomes were also incubated in the presence of human serum at $37^{\circ}C$ for 24 hours. Results: 6-BOC-hydrazinopyridine-3-carboxylic acid was synthesized with 77.3% overall yield. The HYNIC concentration in the PEG-coated liposome dispersion was 1.08 mM. In condition of considering the measured liposomal size of 106 nm, the phospholipid concentration of $77.5\;{\mu}mol/m{\ell}$ and the liposomal particle number of $5.2{\times}10^{14}$ liposomes/ml, it is corresponded to approximate 1,250 nicotinyl hydrazine group per liposome in HYNIC-PEG-liposome. The removal of free $^{99m}Tc$ was not necessary because the labeling efficiency were above 99%. The radiolabeled liposomes maintained 98%, 96% and 99%, respectively, of radioactivity after incubation with transchelators. The radiolabeled liposomes possessed above 90% of the radioactivity in serum. Conclusion: These results suggest that the HYNIC can be synthesized easily and applied in labelling of PEG-liposomes with $^{99m}Tc$.

Optimized Condition of Genomic DNA Extraction and PCR Methods for GMO Detection in Potato (유전자재조합 감자의 검정을 위한 DNA분리 및 PCR검출의 최적조건 탐색)

  • Shin, Weon-Sun;Kim, Myung-Hee
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.591-597
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    • 2003
  • To compare the quality of genomic DNA extracted from potato for PCR detection, four different methods, such as silica-based membrane method, silica-coated bead method, STE solution treatment, and CTAB-phenol/chloroform method, were evaluated. Also, to remove an excessive carbohydrate from the potato, ${\alpha}$- and ${\beta}$-amylase were used individually and in combination. When used both silica-based membrane method and silica-coated bead method combined with enzymes, the genomic DNAs were extracted from the raw potato with high purity for PCR. However, the silica-coated head method combined with enzyme treatment was the most efficient for extraction of the genomic DNA from the frozen fried potatoes. When applied with STE solution, the highly purified DNA was extracted from the raw potatoes without enzyme treatment in adequate yield for PCR. In cases of processed potatoes, such as frozen-fried potato and fabricated potato chips, CTAB-phenol/chloroform method is mostly feasible for DNA extraction and PCR efficacy at high sensitivity. As the results of PCR amplification, 216bp of PCR product was detected on 2% agarose gel electrophoresis, but any amplicons derived from New leaf and New leaf Y gene was not detected in any sample.

Automated Synthesis of [$^{18}F$]Fallypride for Routine Clinical Use (자동합성장치를 이용한 [$^{18}F$]Fallypride의 합성)

  • Park, Jun-Hyung;Moon, Byung-Seok;Lee, Hong-Jin;Lee, Hyo-Jun;Lee, In-Won;Lee, Byung-Chul;Kim, Sang-Eun
    • The Korean Journal of Nuclear Medicine Technology
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    • v.14 no.2
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    • pp.104-109
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    • 2010
  • Purpose: $[^{18}F]$Fallypride plays an effective radiotracer for the study of dopamine $D_2/D_3$ receptor occupancy, neuropsychiatric disorders and aging in humans. This tracer has the potential for clinical use, but automated labeling efficiency showed low radiochemical yields about 5~20% with relatively long labelling time of fluorine-18. In present study, we describe an improved automatic synthesis of [$^{18}F$]Fallypride using different base concentration for routine clinical use. Materials and Methods: Fully automated synthetic process of [$^{18}F$]Fallypride was perform using the TracerLab $FX_{FN}$ synthesizer under various labeling conditions and tosyl-fallypride was used as a precursor. [$^{18}F$]Fluoride was extracted with various concentration of $K_{2.2.2.}/K_2CO_3$ from $^{18}O$-enriched water trapped on the ion exchange cartridge. After azeotropic drying, the labeling reaction proceeded in $CH_3CN$ at $100^{\circ}C$ for 10 or 30 min. The reaction mixture was purified by reverse phase HPLC and collected organic solution was exchanged by tc-18 Sep-Pak for the clinically available solution. Results: The optimal labeling condition of [$^{18}F$]Fallypride in the automatic production was that 2 mg of tosyl-fallypride in acetonitrile (1 mL) was incubated at $100^{\circ}C$ for 10 min with $K_{2.2.2.}/K_2CO_3$ (11/0.8 mg). [$^{18}F$]Fallypride was obtained with high radiochemical yield about $66{\pm}1.4%$ (decay-corrected, n=28) within $51{\pm}1.2$ min including HPLC purification and solid-phase purification for the final formulation. Conclusion: [$^{18}F$]Fallypride was prepared with a significantly improved radiochemical yield with high specific activity and shorten synthetic time. In addition, this automated procedure provides the high reproducibility with no synthesis failures (n=28).

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A Survey on the Consumer Attitude Toward Health Food in Korea (II) -Consumer Perception on Health Foods- (건강식품에 대한 소비자 인식 연구 (II) -건강식품에 관한 소비자 의식구조-)

  • Lee, Eun-Joo;Ro, Seung-Ok;Lee, Cherl-Ho
    • Journal of the Korean Society of Food Culture
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    • v.11 no.4
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    • pp.487-495
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    • 1996
  • The consumer perception on health and food habit, the experience of health food use and the discrimination between health food and drug of Korean consumer were surveyed by using a questionnaire containing 20 items in order to obtain the basic data for the assessment of the benefit and risk of health foods in Korea. A total of 1,000 people over 20 years of age living in Seoul and the vicinities were interviewed and asked to fill out the questionnaire during the period from the October 1995 to the February 1996. Among the 882 answers collected 23 was incomplete data, and 859 answers were used for the statistical analysis by using SAS program. The survey revealed a strong interest of the consumer on health food by showing that more than a half of the subjects (58.8%) had the experience of actual use of health food, and 68.2% believed the effectiveness. What the consumer expect most from health food was to have beneficial effect to maintain overall health condition (59.8%), and the most negative aspect of health food was the overstatement on the effectiveness by the producers (52.1%). The most important source of information for the purchase of health food was the suggestion of friends and relatives (30.6%). Among the health foods registered and regulated by the food law, royal jelly (22.7%), squalene (16.0%), refined fish oil (15.1%), lactic acid bacteria (10.6%) and aloe (8.8%) were relatively well aware. Although 84% of the subjects perceived that health food is different from drug or traditional medicine, the largest percentage of the subject selected ginseng as the most well known type of health food (22.7%) as well as the most well known drug (or traditional medicine) (41.7%). Ginseng was also chosen as the most frequently used health food (17.0%), and vitamin tablets the third (13.0%). The vague definition of health food and unambiguous discrimination of it from medicine by the consumers were problematic for the correct use and reasonable purchasing behavior. The clear definition and proper regulation on the manufacture and distribution of health food, more strict control of labelling and advertisement, and a wide consumer education on health food were recommended.

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