• 제목/요약/키워드: Incubation Characteristics

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Synthesis, Radiolabeling and Gastrin Releasing Peptide Receptor Binding Affinity of a Novel Bombesin Antagonist-Based Peptide, DOTA-Ala(SO3H)-Aminooctanoyl-Gln-Trp-Ala-Val-N methyl Gly-His-Statine-Leu-NH2

  • Lim, Jae Cheong;Choi, Sang Mu;Cho, Eun Ha;Kim, Jin Joo
    • 방사선산업학회지
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    • 제7권2_3호
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    • pp.127-134
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    • 2013
  • Bombesin receptors are overexpressed in many kinds of human tumors. In particular, the gastrin releasing peptide receptor (GRPR) which is also called bombesin receptor subtype 2, has been identified in prostate cancer. In the present study, we developed a bombesin antagonist-based $^{177}Lu$-labeled peptide, $^{177}Lu$-DOTA-$Ala(SO_3H)$-Aminooctanoyl-Gln-Trp-Ala-Val-N methyl Gly-His-Statine-Leu-$NH_2$ (DOTA-sBBNA). DOTA-sBBNA was prepared using a solid phase synthesis method. It was labeled with $^{177}Lu$ by a high radiolabeling yield (>98%), and its Log P value was -2.05. The radiolabeled peptide was highly stable in serum incubation at $37^{\circ}C$ for 48 hr. A competitive displacement of $^{125}I-[Tyr^4]$-Bombesin on the PC-3 human prostate carcinoma cells revealed that the $IC_{50}$ value of the peptide was 6.76 nM indicating a highly nanomolar binding affinity for GRPR. These results suggest that $^{177}Lu$-DOTA-sBBNA can be a potential candidate for targeting prostate cancer, and further studies to evaluate its biological characteristics are needed.

Novel Bombesin Analogues Conjugated with DOTA-Ala(SO3H)-4 aminobenzoic acid and DOTA-Lys(glucose)-4 aminobenzoic acid: Synthesis, Radiolabeling, and Gastrin Releasing Peptide Receptor Binding Affinity

  • Lim, Jae Cheong;Choi, Sang Mu;Cho, Eun Ha;Kim, Jin Joo
    • 방사선산업학회지
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    • 제7권2_3호
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    • pp.191-200
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    • 2013
  • In this study, a novel bombesin (BBN) analogues, DOTA-Ala($SO_3H$)-4 aminobenzoyl-Gln-Trp-Ala-Val-Gly-His-Leu-Met-$NH_2$ (DOTA-sBBN) and DOTA-Lys(glucose)-4 aminobenzoyl-Gln-Trp-Ala-Val-Gly-His-Leu-Met-$NH_2$ (DOTA-gluBBN), were synthesized and radiolabeled, and their binding affinities were evaluated. Peptides were prepared by a solid phase synthesis method and their purities were over 98%. DOTA is the chelating agent for $^{177}Lu$-labeling, and the DOTA-conjugated peptides were radiolabeled with $^{177}Lu$ by a high radiolabeling yield (>98%). The Log P values of DOTA-sBBN and DOTA-gluBBN were -2.20 and -2.79, respectively. 50.41% of $^{177}Lu$-DOTA-sBBN and 72.97% of $^{177}Lu$-DOTA-gluBBN were left undegraded by the serum incubation at $37^{\circ}C$ for 48 hr. A competitive displacement of $^{125}I-[Tyr^4]$-BBN on the PC-3 human prostate carcinoma cells revealed that 50% inhibitory concentration ($IC_{50}$) were 1.46 nM of DOTA-sBBN and 4.67 nM of DOTA-gluBBN indicating a highly nanomolar binding affinity for GRPR. Therefore, it is concluded that $^{177}Lu$-DOTA-sBBN and $^{177}Lu$-DOTA-gluBBN can be potential candidates as a targeting modality for the Gastrin-releasing peptide receptor (GRPR)-over-expressing tumors, and further studies to evaluate their biological and pharmacological characteristics are needed.

Nutritional evaluation of total mixed rations containing rice grain in an in vitro rumen fermentation system

  • Yang, Sung Jae;Kim, Han Been;Moon, Joon Beom;Kim, Na Eun;Park, Joong Kook;Park, Byung Ki;Lee, Se Young;Seo, Jakyeom
    • 농업과학연구
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    • 제45권4호
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    • pp.741-748
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    • 2018
  • This study was conducted to evaluate the nutritional value of total mixed rations (TMR) containing rice grain in an in vitro rumen fermentation system. Three types of grains (corn, wheat, and rice), timothy, and soybean meal (SBM) were used to prepare the experimental TMR: Corn TMR, Wheat TMR, and Rice TMR. The rumen fermentation characteristics of all the experimental TMRs were evaluated by an in vitro anaerobic system using rumen fluid for 24 and 48 h. The digestibility of the nutrients (dry matter [DM], crude protein [CP], and neutral detergent fiber [NDF]), pH, ammonia ($NH_3-N$), and volatile fatty acids (VFA) were determined. Rice TMR showed a higher DM digestibility than that of the Corn TMR at 48 h (p < 0.05). In all treatments, the CP digestibility was more than 80% at 48 h, but no significant differences were observed among the treatments. The NDF digestibility tended to be the lowest in the Wheat TMR (p = 0.06), and the pH tended to be the lowest in the Rice TMR (p = 0.09) among the treatments for the 48 h incubation. The Wheat TMR had the highest $NH_3-N$ concentration among the treatments (p < 0.01). Rice TMR had a lowest total VFA concentration among the treatments (p = 0.05) at 24 h, but no significant differences were observed at 48 h. Based on this in vitro result, it was considered that a rice grain has the potential to replace conventional grain ingredients when the TMR was formulated.

A study on comparative feeding value of corn flakes according to temperature and retention time in the pressurized steam chamber

  • Ahn, Jun Sang;Shin, Jung Suh;Kim, Min Ji;Son, Gi Hwal;Kwon, Eung Gi;Shim, Jae Yoon;Kim, Il Young;Cho, Sung Myoun;Cho, Sang Rae;Park, Byung Ki
    • Journal of Animal Science and Technology
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    • 제61권3호
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    • pp.170-181
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    • 2019
  • This study aimed to investigate the effects of temperature and retention time of the pressurized steam chamber on the ruminal fermentation characteristics and nutrient degradability of corn flakes in three Korean native Hanwoo cows and three Holstein cows implanted with a ruminal fistula. Corn kernels were categorized into 13 groups based on the chamber temperature (range, $100^{\circ}C-116^{\circ}C$) and retention time (range, 700-950 s). The pH value was lowest in T1 regardless of breed. Propionate concentration was the highest in T2 (p < 0.05). Total-volatile fatty acid (VFA) concentration was slightly but not significantly greater in T2 than in other conditions. Dry matter (p < 0.05), starch, and crude protein (p < 0.05) degradability were the highest in T1. At different incubation times and with different breeds, dry matter, starch, and crude protein degradability of corn flakes were the highest in T1. Thus, the present results indicate that the optimal temperature and retention time of the pressurized steam chamber should be $100^{\circ}C-105^{\circ}C$ and 700-720 s.

Nutrient and ruminal fermentation profiles of Camellia seed residues with fungal pretreatment

  • Yang, Chunlei;Chen, Zhongfa;Wu, Yuelei;Wang, Jiakun
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권3호
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    • pp.357-365
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    • 2019
  • Objective: The experiment was conducted to evaluate the effects of four fungal pretreatments on the nutritional value of Camellia seed residues, and to evaluate the feeding value of pretreated Camellia seed residues for ruminants. Methods: Camellia seed residues were firstly fermented by four lignin degrading fungi, namely, Phanerochaete chrysosporium (P. chrysosporium)-30942, Trichoderma koningiopsis (T. koningiopsis)-2660, Trichoderma aspellum (T. aspellum)-2527, or T. aspellum-2627, under solid-state fermentation (SSF) conditions at six different incubation times. The nutritional value of each fermented Camellia seed residues was then analyzed. The fermentation profiles, organic matter degradability and metabolizable energy of each pre-treated Camellia seed residue were further evaluated using an in vitro rumen fermentation system. Results: After 5 days of fermentation, P. chrysosporium-30942 had higher degradation of lignin (20.51%), consumed less hemicellulose (4.02%), and the SSF efficiency reached 83.43%. T. koningiopsis-2660 degraded more lignin (21.54%) and consumed less cellulose (20.94%) and hemicellulose (2.51%), the SSF efficiency reached 127.93%. The maximum SSF efficiency was 58.18% for T. aspellum-2527 and 47.61% for T. aspellum-2627, appeared at 30 and 15 days respectively. All the fungal pretreatments significantly improved the crude protein content (p<0.05). The Camellia seed residues pretreated for 5 days were found to possess significantly increased organic matter degradability, volatile fatty acid production and metabolizable energy (p<0.05) after the treatment of either P. chrysosporium-30942, T. koningiopsis-2660 or T. aspellum-2527. The fungal pretreatments did not significantly change the rumen fermentation pattern of Camellia seed residues, with an unchanged ratio of acetate to propionate. Conclusion: The fungi showed excellent potential for the solid-state bioconversion of Camellia seed residues into digestible ruminant energy feed, and their shorter lignin degradation characteristics could reduce loss of the other available carbohydrates during SSF.

Effect of flaking on the digestibility of corn in ruminants

  • Kang, Hamin;Lee, Mingyung;Jeon, Seoyoung;Lee, Sang Moon;Lee, Ju Hwan;Seo, Seongwon
    • Journal of Animal Science and Technology
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    • 제63권5호
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    • pp.1018-1033
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    • 2021
  • In this study, we aimed to assess the effect of flaking on the nutrient digestibility of corn grain in ruminants. In this regard, in vitro rumen fermentation, in situ rumen degradability, and in vivo metabolic experiments were performed. The automated gas production technique was used for the in vitro fermentation experiments. Six types of corn flakes with various degrees of gelatinization (32%, 41%, 48%, 66%, 86%, and 89%) were ground and incubated in rumen fluid to measure rumen fermentation characteristics and digestion rate. The in situ degradability of ground corn, whole corn, and corn flakes with 62% and 66% gelatinization was measured by incubation in the rumen of two cannulated Holstein cows. In vivo metabolic experiments were performed using 12 crossbred goats (29.8 ± 4.37 kg) using a 3 × 3 Latin square design. The dietary treatments consisted of ground corn and flaked corn with 48% or 62% gelatinization. In vitro experiments showed that as the degree of gelatinization increased, the digestion rate increased linearly, while the discrete lag time decreased linearly (p < 0.05). The effective rumen dry matter degradability, determined by in situ fermentation, was 37%p lower in corn flakes than ground corn, assuming a passage rate of 6%/h (p < 0.01), and there was no difference between the two flakes. In the in vivo experiment, there was no difference in dry matter intake, average daily gain, feed efficiency, and nitrogen utilization among the treatment groups (p > 0.05); however, the crude fat digestibility was lower for corn flakes than for ground corn (p < 0.05). To summarize, the rate of fermentation of corn flakes increased as the degree of gelatinization increased. However, non-ground corn flakes had lower rumen digestibility and did not improve in vivo apparent nutrient digestibility, compared with ground corn. In contrast to the assumption that flaked corn provides more energy to ruminant animals than ground corn, we conclude that the digestibility and energy value of corn flakes are lower than those of ground corn if mastication does not sufficiently reduce the particle size of corn flakes.

일반 유기물 항목과 분광특성을 이용한 한강수계 내 난분해성 물질 지표 제시 (Development of Estimation Indices for Refractory Organic Matter in the Han-River Basin using Organic Matter Parameters and Spectroscopic Characteristics)

  • 이보미;이태환;허진
    • 한국물환경학회지
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    • 제27권5호
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    • pp.625-633
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    • 2011
  • A long-term water quality monitoring in the Han River Basin reveals a consistent increasing trend for the concentration of refractory organic matter (R-OM) in major monitoring sites of the watershed. Because the determination of R-OM concentrations typically requires a long time of microbial incubation, it is essential to present the estimation indices for R-OM for an efficient watershed management. In this study, a number of surface water samples were classified into three groups, each of which were collected from Lake Paldang, rivers at rain and non-rain events, respectively. The corresponding R-OM concentrations were correlated with biochemical oxygen demand (BOD), chemical oxygen demand (COD), and total organic carbon (TOC) concentrations as well as ultraviolet and fluorescence intensities of the filtered samples. Among the traditional organic matter parameters, TOC exhibited the highest correlation coefficient with the R-OM concentrations regardless of the types of the sample groups. The equations for conversing TOC into R-OM concentrations were finally suggested as $0.43{\times}TOC+1.12$, $0.44{\times}TOC+0.61$, $0.24{\times}TOC+1.28$ for river samples at rain and non-rain events, and lake samples, respectively. TOC-BOD(C), the values of the TOC concentrations subtracted by carbon-converted BOD concentrations, was a good index for estimating the absolute concentrations of R-OM. UV absorbance at 254 nm was well correlated with R-OM concentrations of river samples while fluorescence intensities at 350 nm showed an excellent relationship with R-OM concentration of the lake samples. Our results suggests that simple spectroscopic parameters could be applied for in-situ monitoring tool techniques in watersheds.

High Prevalence of Listeria monocytogenes in Smoked Duck: Antibiotic and Heat Resistance, Virulence, and Genetics of the Isolates

  • Park, Eunyoung;Ha, Jimyeong;Oh, Hyemin;Kim, Sejeong;Choi, Yukyung;Lee, Yewon;Kim, Yujin;Seo, Yeongeun;Kang, Joohyun;Yoon, Yohan
    • 한국축산식품학회지
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    • 제41권2호
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    • pp.324-334
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    • 2021
  • This study aimed at determining the genetic and virulence characteristics of the Listeria monocytogenes from smoked ducks. L. monocytogenes was isolated by plating, and the isolated colonies were identified by PCR. All the obtained seven L. monocytogenes isolates possessed the virulence genes (inlA, inlB, plcB, and hlyA) and a 385 bp actA amplicon. The L. monocytogenes isolates (SMFM2018 SD 1-1, SMFM 2018 SD 4-1, SMFM 2018 SD 4-2, SMFM 2018 SD 5-2, SMFM 2018 SD 5-3, SMFM 2018 SD 6-2, and SMFM 2018 SD 7-1) were inoculated in tryptic soy broth (TSB) containing 0.6% yeast extract at 60℃, followed by cell counting on tryptic soy agar (TSA) containing 0.6% yeast extract at 0, 2, 5, 8, and 10 min. We identified five heat resistant isolates compared to the standard strain (L. monocytogenes ATCC13932), among which three exhibited the serotype 1/2b and D-values of 5.41, 6.48, and 6.71, respectively at 60℃. The optical densities of the cultures were regulated to a 0.5 McFarland standard to assess resistance against nine antibiotics after an incubation at 30℃ for 24 h. All isolates were penicillin G resistant, possessing the virulence genes (inlA, inlB, plcB, and hlyA) and the 385-bp actA amplicon, moreover, three isolates showed clindamycin resistance. In conclusion, this study allowed us to characterize L. monocytogenes isolates from smoked ducks, exhibiting clindamycin and penicillin G resistance, along with the 385-bp actA amplicon, representing higher invasion efficiency than the 268-bp actA, and the higher heat resistance serotype 1/2b.

A report of 28 unrecorded bacterial species in Korea, isolated from freshwater and sediment of the Han River watershed in 2020

  • Kim, Mirae;Song, Jaeho;Yu, Dabin;Kim, Younghoo;Bae, Seok Hwan;Park, Miri S.;Lim, Yeonjung;Cho, Jang-Cheon
    • Journal of Species Research
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    • 제10권3호
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    • pp.227-236
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    • 2021
  • To obtain unrecorded freshwater bacterial species in Korea, water and sediment samples were collected from streams, lakes, and wetland of the Han River watershed in 2020. Approximately 800 bacterial strains were isolated on R2A agar after aerobic or anaerobic incubation, and identified using 16S rRNA gene sequences. A total of 28 strains, with ≥98.7% 16S rRNA gene sequence similarity with validly published bacterial species but not reported in Korea, were determined to be unrecorded bacterial species in Korea. The unrecorded bacterial strains were phylogenetically diverse and belonged to four phyla, eight classes, 13 orders, 19 families, and 25 genera. The unreported species were assigned to Acetobacter, Alsobacter, Mesorhizobium, Prosthecomicrobium, and Microvirga of the class Alphaproteobacteria; Vogesella, Formosimonas, Aquincola, Massilia, Acidovorax, and Brachymonas of the class Betaproteobacteria; Pseudoxanthomonas, Thermomonas, Lysobacter, Enterobacter, Kosakonia, and Acinetobacter of the class Gammaproteobacteria; Sulfuricurvum of the class Epsilonproteobacteria; Mycolicibacterium, Agromyces, Phycicoccus, and Microbacterium of the class Actinobacteria; Paenibacillus of the class Bacilli; Clostridium of the class Clostridia; and Flavobacterium of the class Flavobacteriia. The details of the unreported species, including Gram reaction, colony and cell morphology, biochemical characteristics, and phylogenetic position are also provided in the description of the strains.

Optimized pretreatment conditions for the environmental DNA (eDNA) analysis of Apostichopus japonicus

  • Kang, Yu-An;Lee, Soo Rin;Kim, Eun-Bi;Park, Sang Un;Lim, Sang Min;Andriyono, Sapto;Kim, Hyun-Woo
    • Fisheries and Aquatic Sciences
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    • 제25권5호
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    • pp.264-275
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    • 2022
  • A non-destructive environmental DNA protocol for the genetic analysis of sea cucumber (Apostichopus japonicus) resources DNA was established. Among the several commercial DNA extraction kits, the DNeasy® Plant Mini Kit was selected as the best choice to obtain the high-quality genomic DNAs from the mucous sea cucumber. As the temperature and incubation time increased, the amount of extracted environmental DNA was also large, but it was judged that the increased amount did not affect as much as 2-3 times. Therefore, these conditions were not considered to be the main factors to consider in actual environmental DNA extraction. However, the amount of seawater relative to the size of the sample was judged as a major consideration, and a sufficient amount of environmental DNA for analysis was secured when stored within 1 min while stirring the volume of seawater corresponding to the total sea cucumber weight (g). In securing the environmental DNA of sea cucumbers, the mortality rate of sea cucumbers in all experiments was 0, and it was judged that the effects of sea cucumbers were not significant through this treatment. Through the results of this study, sea cucumber DNA research, which has been conducted in a destructive method, can be conducted non-destructively through environmental DNA analysis. Through this study, we have secured a standard protocol that can successfully extract the sea cucumber DNA through environmental DNA. It is not only excellent in terms of time and cost of traditional DNA analysis method currently used, but it is completely non-destructive in the ecosystem of the survey area. It is believed that the system can be transformed in a way that does not affect it. However, it is thought that various standard protocols should be established considering the characteristics of each type.