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PET Imaging of Click-engineered PSMA-targeting Immune Cells in Normal Mice

  • Hye Won Kim;Won Chang Lee;In Ho Song;Hyun Soo Park;Sang Eun Kim
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.8 no.2
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    • pp.53-61
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    • 2022
  • This study aimed to increase the targeting ability against PSMA in cell therapy using metabolic glycoengineering and biorthogonal chemistry and to visualize cell trafficking using PET imaging. Cellular membranes of THP-1 cells were decorated with azide(-N3) using Ac4ManNAz by metabolic glycoengineering. Engineered THP-1 cells were conjugated with DBCO-bearing fluorophore (ADIBO-Cy5.5) for 1 h at different concentrations and analyzed by confocal fluorescence microscopy and flow cytometry. For PSAM ligand conjugation to THP-1 cells, Ac4ManNAz treated THP-1 cells were incubated with DBCO-PSMA ligand (ADIBO-GUL) at a final concentration with 100 µM for 1 h. To evaluate the effect on cell recognition, PSMA ligand conjugated THP-1 cells(as effectors) were co-cultured with PSMA positive 22RV1 (as target cells) at 3 : 1 a effector-to-target cell (E/T) ratio. The interaction between THP-1 and 22RV1 was monitored by confocal fluorescence microscopy. For preparing the radiolabeled THP-1, the cells were treated at the activity of ~ 740 kBq of [89Zr]Zr(oxinate)4/5 × 106 cells. Radiolabeled cells were analyzed for determination of cell-associated radioactivity by gamma counting and viability using MTS assay. In the cytotoxicity assay, THP-1 cells did not have any cytotoxicity even when the Ac4ManNAz concentration was 100 µM. In confocal microscopy and flow cytometry, THP-1 cells were efficiently labeled ADIBO-Cy5.5 in a dose-dependent manner, and the dose of 100 µM was the optimal concentration for the following experiments. The clusters of PSMA ligand-conjugated THP-1 cells and 22RV1 cells were identified, indicating cell-cell recognition over the cell surface between two types of cells. Cell radiolabeling efficiency was 54.5 ± 17.8%. THP-1 labeled with 0.09 ± 0.03 Bq/cell showed no significant cytotoxicity compared to unlabeled THP-1 up to 7 days. We successfully demonstrated that Ac4ManNAz treated cells were efficiently conjugated with ADIBO-GUL for preparing the PSMA-targeting cells, and [89Zr]Zr(oxinate)4 could be used to label cells without toxicity. It suggested that PSMA-ligand conjugated cell therapy could be improved cell targeting and be monitored by PET imaging.

Effect of New Herbal Mixture, HT008 on Arthritis Rat Model (한약복합물 HT008이 흰쥐 관절염 모델에 미치는 효과)

  • Lim, Dong-Wook;Kim, Mi-Yeon;Song, Mi-Kyung;Kim, Ji-Young;Lee, Dong-Hun;Park, Ju-Yeon;Bu, Young-Min;Kim, Ho-Cheol
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.197-204
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    • 2009
  • Objectives : This study was conducted to investigate the effect of HT008, a multi-herbal mixture consisting of 3 herbs, Eleutherococcus senticosus, Angelica sinensis, and Scutellaria baicaleinsis on arthritic model in rats. Methods : The anti-inflammatory and analgesic activities were observed by utilizing the following models: carrageenan-induced edema of the hind paw of rats, acetic acid-induced writhing response in mice. The perimeter of the paw was measured before injection and then at 1, 2, 4, 6 h after injection of 1% $\lambda$-carrageenan. The HT008 at five dose levels (10, 30, 100, and 300 mg/kg) and distilled water given 30 min to treatment groups and control group, before $\lambda$-carrageenan injection. In the writhing test, the mice received 0.7% acetic acid solution in normal saline injected intraperitoneally at a dose of 10 ml/kg. The number of writhes was counted staring 10 min after injection. Results : HT008 at four dose levels (30, 100, and 300 mg/kg) significantly decreased the carrageenan-induced rat paw edema perimeter. E. senticosus and S. baicaleinsis extracts reduced acetic acid-induced writhing response in mice. Also A. sinensis extracts significantly decreased the carrageenan-induced rat paw edema perimeter. Conclusions : These results show that HT008, a multi-herbal mixture has both anti-inflammatory activity and analgesic effects in vivo arthritic model, and suggest that HT008 could be a good therapy to treat human osteoarthritis.

Experimental Study on the Adsorption Characteristics of Methane Gas Considering Coalbed Depth in Coalbed Methane Reservoirs (석탄층 메탄가스 저류층에서 탄층 심도를 고려한 메탄가스의 흡착 특성에 관한 실험 연구)

  • Chayoung Song;Dongjin Lee;Jeonghwan Lee
    • Journal of the Korean Institute of Gas
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    • v.27 no.2
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    • pp.39-48
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    • 2023
  • This study presents the experimental results to measure the adsorption amount of methane gas by coal according to the conditions of a coalbed methane (CBM) reservoir. Adsorbed gas to coal seam particles was measured under reservoir conditions (normal pressure ~ 1,200 psi pressure range, temperature range15 ~ 45℃) using coal samples obtained from random mines in Kalimantan Island, North Indonesia. The obtained amount of absolute adsorbed gas was applied to triangular with linear interpolation to calculate the maximum amount of adsorbed gas according to temperature and pressure change, at which no experiment was performed. As a result, it was revealed that the amount of adsorbed gas to coal particles increased as the pressure increased and temperature decreased, but the increase of the amount of adsorbed gas decreased at more than an appropriate depth(1,000 ft). In the cleat permeability and cleat porosity for each depth of the coal bed considering the effective stress, the cleat permeability was 28.86 ~ 46.81 md, and the cleat porosity was 0.83 ~ 0.98%. This means that the gas productivity varies significantly with the depth because the reduction of the permeability according to the depth in the coal seam is significant. Therefore, a coalbed depth should be considered essential when designing the spacing of production wells in a coalbed methane reservoir in further study.

Effects of different stocking density and various phytogenic feed additives dosage levels on growing-finishing pigs

  • Hyun Ah Cho;Min Ho Song;Ji Hwan Lee;Han Jin Oh;Yong Ju Kim;Jae Woo An;Se Yeon Chang;Young Bin Go;Dong Cheol Song;Seung Yeol Cho;Dong Jun Kim;Mi Suk Kim;Hae Ryoung Park;Hyeun Bum Kim;Jin Ho Cho
    • Journal of Animal Science and Technology
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    • v.65 no.3
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    • pp.535-549
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    • 2023
  • This study was to investigate the effects of different phytogenic feed additives (PFA) dosage levels in growing- finishing pigs stressed by high stocking density. A total of 72 mix sexed 12 weeks growing pigs ([Landrace × Yorkshire] × Duroc) with initial body weight (BW) of 49.28 ± 4.58 kg were used for 8 weeks. There were 3 replicate pens in each treatment group, with 3 pigs per pen. The dietary treatment groups consisted of basal diets in animal welfare density (negative control [NC]), basal diet in high stocking density (positive control [PC]), PC + 0.04% essential oil (ES1), PC + 0.08% essential oil (ES2), PC + 0.10% bitter citrus extract & essential oil (CES1), PC + 0.20% bitter citrus extract & essential oil (CES2), PC + 0.05% grape pomace extract (GP1), PC + 0.10% grape pomace extract (GP2). The reduction of space allowance decreased (p < 0.05) average daily gain, feed efficiency, and digestibility of dry matter, crude protein, and gross energy. Also, the fecal score of PC groups increased (p < 0.05) compared with other groups. Basic behaviors (feed intake, standing, lying) were inactive (p < 0.05) and singularity behavior (biting) was increased (p < 0.10) under high stocking density. There was no difference in blood profile. However, the supplementation of PFA alleviated the negative effects such as reduced growth performance, nutrient digestibility, and some increasing stress indicators in th blood (cortisol) and animal behavior (biting). In conclusion, the negative effect of high stocking density was most effectively mitigated by the normal dosage of the mixture of bitter citrus extract and essential oil additive (CES1).

Effects of Nutrient Solution Strength on Growth and Nutrient Element Concentrations of Leaf Lettuce by Hydroponic Culture under Artificial Light (인공광을 이용한 수경재배에서 배양액 농도가 상추의 생장과 배양액 양분 농도에 미치는 영향)

  • Kim, D.E.;Lee, W.Y.;Heo, J.W.;Lee, G.I.;Kang, D.H.;Woo, Y.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.19 no.1
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    • pp.5-14
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    • 2017
  • This study was conducted to investigate the effects of nutrient solution strength on growth and nutrient element concentrations in leaf lettuce (Lactuca sativa L. cv. 'Dduksum') by hydroponic culture under fluorescent lamp and LED. Leaf lettuce were grown in closed hydroponic cultivation systems supplied with 1/2, 1 and 2 strength of nutrient solution recommended by horticultural experiment station in Japan. The growth of 'Dduksum' was highest in the 2 strength of standard nutrient solution. The amount of nutrient element in the recycled nutrient solution was higher at 2, 1 and 1/2 strength of nutrient solution. The concentration of NO3-N, Ca2+, Mg2+ in the recycled nutrient solutions increased in 1 and 2 strength of nutrient solution but that of NH4-N decreased gradually in 1/2 and 1 strength of nutrient solution. The concentration of K, Ca, Mg in leaf lettuce was maintained in the normal range, whereas the concentration of phosphorous was 1.3 to 1.6%, which was higher than proper range. As the concentration of NH4-N decreases gradually in all the treatments, it is necessary to raise the rate of NH4-N or add it.

Effects of different levels of organic chromium and selenomethionine cocktails in broilers

  • Jaewoo An;Younggwang Kim;Minho Song;Jungseok Choi;Hanjin Oh;Seyeon Chang;Dongcheol Song;Hyunah Cho;Sehyun Park;Kyeongho Jeon;Yunhwan Park;Gyutae Park;Sehyuk Oh;Yuna Kim;Nayoung Choi;Jongchun Kim;Hyeunbum Kim;Jinho Cho
    • Journal of Animal Science and Technology
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    • v.65 no.6
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    • pp.1226-1241
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    • 2023
  • Selenium (Se) is an essential trace mineral that plays an important role in physiological processes by regulating the antioxidant defense system and enhancing immunity. Chromium is an essential mineral involved in carbohydrate and lipid metabolism and also plays a role in maintaining normal insulin function. Based on these advantages, we hypothesized that the addition of selenomethionine (SeMet) and organic chromium (OC) to broiler diets would increase Se deposition, antioxidant capacity and immune response in meat. Therefore, this study analyzed the effects of OC and SeMet on growh performance, nutrients digestibility, blood profiles, intestinal morphology, meat quality characteristics, and taxonomic analysis of broilers. A total of 168 one-day-old broiler chicken (Arbor Acres) were randomly allotted to 3 groups based on the initial body weight of 37.33 ± 0.24 g with 7 replicate per 8 birds (mixed sex). The experiments period was 28 days. Dietary treatments were folloewd: Basal diets based on corn-soybean meal (CON), basal diet supplemented with 0.2 ppm OC and 0.2 ppm SeMet (CS4), and basal diet supplemented with 0.4 ppm OC and 0.4 ppm SeMet (CS8). Supplementation of OC and SeMet did not affect on growth performance, nutrient digestibility. However, CS8 supplementation increased in duodenum villus height and villus height : crypt depth, and increased in breast meat Se deposition. In addition, CS8 group showed higher uric acid and total antioxidant status than CON group. Taxonomic analysis at phylum level revealed that Proteobacteria and Firmicutes of CS4 and CS8 were lower than CON group. In genus level, the relative abundance of fecal Lactobacillus and Enterococcus of CS4 and CS8 groups were higher than CON group. In short, 0.4 ppm OC and 0.4 ppm SeMet supplementation to broiler diet supporitng positive gut microbiome change, also enhancing antioxidant capacity, and Se deposition in breast meat.

Muscle Fiber, Connective Tissue and Meat Quality Characteristics of Pork from Low Birth Weight Pigs as Affected by Diet-Induced Increased Fat Absorption and Preferential Muscle Marbling

  • Bimol C. Roy;Patience Coleman;Meghan Markowsky;Kun Wang;Yongbo She;Caroline Richard;Spencer D. Proctor;Heather L. Bruce
    • Food Science of Animal Resources
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    • v.44 no.1
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    • pp.51-73
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    • 2024
  • This study investigated how birth weight differences in piglets affected carcass and muscle fiber properties as well as meat quality at slaughter. Within litters, piglets were grouped according to their birth weight as either normal (NBW; 1.62-1.73 kg) or low (LBW; 1.18-1.29 kg). At 5 weeks of age, NBW piglets were randomly transitioned to control (C) or isocaloric high fat diets derived from non-dairy (HF), while LBW piglets were randomly transitioned to high fat diets derived from non-dairy (HF) or dairy sources (HFHD). Piglets were reared in individual pens under standardized housing and feeding conditions. Live weight was recorded weekly, and pigs were slaughtered at 12 weeks of age. Hot carcass weights, dressing percentages, lean meat yield, and primal cut proportions were determined. The m. longissimus thoracis was collected from the right side of the carcass for measurement of physical and chemical properties of meat and muscle fiber characteristics. Results indicated that LBW pigs compensated for their live weight compared to NBW pigs at 6 weeks of age. The mean muscle fiber diameter of LBW-HFHD group is significantly higher than NBW-C and NBW-HF group, and the type I muscle fiber diameter is significantly higher than NBW-C group. Dairy fat inclusion in LBW pig diet reduced carcass back fat thickness. This increased the calculated lean meat yield to be comparable to that of NBW pigs fed a commercial diet. Incorporating dairy-sourced high-fat into LBW pigs' diets appears to be an effective strategy for producing carcasses equivalent to NBW pigs.

Automatic Recognition of Pitch Accent Using Distributed Time-Delay Recursive Neural Network (분산 시간지연 회귀신경망을 이용한 피치 악센트 자동 인식)

  • Kim Sung-Suk
    • The Journal of the Acoustical Society of Korea
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    • v.25 no.6
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    • pp.277-281
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    • 2006
  • This paper presents a method for the automatic recognition of pitch accents over syllables. The method that we propose is based on the time-delay recursive neural network (TDRNN). which is a neural network classifier with two different representation of dynamic context: the delayed input nodes allow the representation of an explicit trajectory F0(t) along time. while the recursive nodes provide long-term context information that reflects the characteristics of pitch accentuation in spoken English. We apply the TDRNN to pitch accent recognition in two forms: in the normal TDRNN. all of the prosodic features (pitch. energy, duration) are used as an entire set in a single TDRNN. while in the distributed TDRNN. the network consists of several TDRNNs each taking a single prosodic feature as the input. The final output of the distributed TDRNN is weighted sum of the output of individual TDRNN. We used the Boston Radio News Corpus (BRNC) for the experiments on the speaker-independent pitch accent recognition. π 1e experimental results show that the distributed TDRNN exhibits an average recognition accuracy of 83.64% over both pitch events and non-events.

Efficient Anti-Tumor Immunotherapy Using Tumor Epitope-Coated Biodegradable Nanoparticles Combined With Polyinosinic-Polycytidylic Acid and an Anti-PD1 Monoclonal Antibody

  • Sang-Hyun Kim;Ji-Hyun Park;Sun-Jae Lee;Hee-Sung Lee;Jae-Kyung Jung;Young-Ran Lee;Hyun-Il Cho;Jeong-Ki Kim;Kyungjae Kim;Chan-Su Park;Chong-Kil Lee
    • IMMUNE NETWORK
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    • v.22 no.5
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    • pp.42.1-42.20
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    • 2022
  • Vaccination with tumor peptide epitopes associated with MHC class I molecules is an attractive approach directed at inducing tumor-specific CTLs. However, challenges remain in improving the therapeutic efficacy of peptide epitope vaccines, including the low immunogenicity of peptide epitopes and insufficient stimulation of innate immune components in vivo. To overcome this, we aimed to develop and test an innovative strategy that elicits potent CTL responses against tumor epitopes. The essential feature of this strategy is vaccination using tumor epitope-loaded nanoparticles (NPs) in combination with polyinosinic-polycytidylic acid (poly-IC) and anti-PD1 mAb. Carboxylated NPs were prepared using poly(lactic-co-glycolic acid) and poly(ethylene/maleic anhydride), covalently conjugated with anti-H-2Kb mAbs, and then attached to H-2Kb molecules isolated from the tumor mass (H-2b). Native peptides associated with the H-2Kb molecules of H-2Kb-attached NPs were exchanged with tumor peptide epitopes. Tumor peptide epitope-loaded NPs efficiently induced tumor-specific CTLs when used to immunize tumor-bearing mice as well as normal mice. This activity of the NPs significantly was increased when co-administered with poly-IC. Accordingly, the NPs exerted significant anti-tumor effects in mice implanted with EG7-OVA thymoma or B16-F10 melanoma, and the anti-tumor activity of the NPs was significantly increased when applied in combination with poly-IC. The most potent anti-tumor activity was observed when the NPs were co-administered with both poly-IC and anti-PD1 mAb. Immunization with tumor epitope-loaded NPs in combination with poly-IC and anti-PD1 mAb in tumor-bearing mice can be a powerful means to induce tumor-specific CTLs with therapeutic anti-tumor activity.

Minimum area for circular isolated footings with eccentric column taking into account that the surface in contact with the ground works partially in compression

  • Inocencio Luevanos-Soto;Arnulfo Luevanos-Rojas;Victor Manuel Moreno-Landeros;Griselda Santiago-Hurtado
    • Coupled systems mechanics
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    • v.13 no.3
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    • pp.201-217
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    • 2024
  • This study aims to develop a new model to obtain the minimum area in circular isolated footings with eccentric column taking into account that the surface in contact with the ground works partially in compression, i.e., a part of the contact area of the footing is subject to compression and the other there is no pressure (pressure zero). The new model is formulated from a mathematical approach based on a minimum area, and it is developed by integration to obtain the axial load "P", moment around the X axis "Mx" and moment around the Y axis "My" in function of σmax (available allowable soil pressure) R (radius of the circular footing), α (angle of inclination where the resultant moment appears), y0 (distance from the center of the footing to the neutral axis measured on the axis where the resultant moment appears). The normal practice in structural engineering is to use the trial and error procedure to obtain the radius and area of the circular footing, and other engineers determine the radius and area of circular footing under biaxial bending supported on elastic soils, but considering a concentric column and the contact area with the ground works completely in compression. Three numerical problems are given to determine the lowest area for circular footings under biaxial bending. Example 1: Column concentric. Example 2: Column eccentric in the direction of the X axis to 1.50 m. Example 3: Column eccentric in the direction of the X axis to 1.50 m and in the direction of the Y axis to 1.50 m. The new model shows a great saving compared to the current model of 44.27% in Example 1, 50.90% in Example 2, 65.04% in Example 3. In this way, the new minimum area model for circular footings will be of great help to engineers when the column is located on the center or edge of the footing.