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Screening of key miRNAs related with the differentiation of subcutaneous adipocytes and the validation of miR-133a-3p functional significance in goats

  • Xin, Li;Hao, Zhang;Yong, Wang;Yanyan, Li;Youli, Wang;Jiangjiang, Zhu;Yaqiu, Lin
    • Animal Bioscience
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    • v.36 no.1
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    • pp.144-155
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    • 2023
  • Objective: Adipocyte differentiation is regulated by a variety of functional genes and noncoding RNAs. However, the role of miRNAs in lipid deposition of goat white adipose tissue is still unclear. Therefore, this study revealed the miRNA expression profile in goat subcutaneous adipocytes by sRNA-seq. Methods: The miRNA expressed in goat subcutaneous preadipocytes and the mature adipocytes were sequenced by sRNA-seq. The differentially expressed miRNAs (DEm) were screened and gene ontology (GO) and Kyoto encyclopedia for genes and genomes (KEGG) analyses were performed. Gain-of-function and loss-of-function combined with oil red O staining, Bodipy staining, and quantitative reverse-transcription polymerase chain reaction (qPCR) were utilized to determine the effect of miR-133a-3p on adipocyte differentiation. Results: A total of 218 DEm were screened out. The target genes of these DEm were significantly enriched in GO items such as biological regulation and in KEGG terms such as FAK signaling pathway and MAPK signaling pathway. qPCR verified that the expression trend of miRNA was consistent with miRNA-seq. The gain-of-function or loss-of-function of miR-133a-3p showed that it promoted or inhibited the accumulation of lipid droplets, and CCAAT enhancer binding protein α (C/EBPα) and C/EBPβ were extremely significantly up-regulated or down-regulated respectively (p<0.01), the loss-of-function also led to a significant down-regulation of peroxisome proliferator activated receptor gamma (PPARγ) (p<0.01). Conclusion: This study successfully identified miRNAs expression patterns in goat subcutaneous adipocytes, and functional identification indicates that miR-133a-3p is a positive regulator of the differentiation process of goat subcutaneous adipocytes. Our results lay the foundation for the molecular mechanism of lipid deposition in meat-source goats from the perspective of miRNA.

Effect of Feeding with Different Source of Carbohydrate and Fiber on Carbohydrate and Lipid Metabolism in Type 2 Diabetic Rats (탄수화물의 급원과 식이섬유의 종류를 달리한 식이가 제 2형 당뇨 쥐의 당대사 및 지질대사에 미치는 영향)

  • Kwon, Sang-Hee;Jeong, Hye-Jin;Shim, Jee-Ae;Son, Young-Ae;Kim, Mi-Kyung
    • Journal of the Korean Society of Food Culture
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    • v.22 no.1
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    • pp.157-165
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    • 2007
  • This study was designed to evaluate the effects of fructose(F) or sucrose(S) and guar gum intake on carbohydrate and lipid metabolism in 15-week-old male Goto-Kakizaki(GK) rats. Fifty rats were randomly assigned to 5 groups which were different in carbohydrate(25% of carbohydrate) and fiber(5% w/w) sources. The carbohydrate(CHO) sources of each group were comstarch(control group, 100% of CHO), fructose with cellulose(F), fructose with guar gum(FG), sucrose with cellulose(S), and sucrose with guar gum(SG). Each group was fed exterimental diet for 4 weeks. We measured food intake, body weight gain, adipose tissues weight and organs weight. We conducted oral glucose tolerance test(OGTT) and measured plasma insulin concentration to examine carbohydrate metabolism. To evaluate lipid metabolism, we measured the lipid profile of plasma, liver and feces. Food intake and weight gain of FG or SG groups tended to be less than those of F or S groups. Perirenal and epididymal fat pad weights of SG group were significantly lower than those of S group and those of FG group tended to be lower than those of F group. In OGTT, blood glucose values of F or S groups were significantly higher than those of C group, and FG or SG groups tended to be lower than those of F or S groups during the experimental time. The area under the curve(AUC) of C group was significantly highest among the groups, AUC and plasma insulin concentration of FG or SG groups tended to be lower than those of F or S groups. Plasma and hepatic triglyceride (TG) of FG and SG groups were significantly lower than those of F and S groups, plasma and hepatic total lipid(TL) and total cholesterol(TC) of FG and SG groups tended to be lower than those of F and S groups. Fecal TL, TG and TC of FG or SG groups tended to be higher than those of F and S groups. In conclusion, intake of guar gum should improve carbohydrate and lipid metabolism in partial substitution of fructose or sucrose for cornstarch in GK rats.

Exploration of nutritional and bioactive peptide properties in goat meat from various primal cuts during in vitro gastrointestinal digestion and absorption

  • Pichitpon Luasiri;Papungkorn Sangsawad;Jaksuma Pongsetkul;Pramote Paengkoum;Chatsirin Nakharuthai;Saranya Suwanangul;Sasikan Katemala;Narathip Sujinda;Jukkrapong Pinyo;Jarunan Chainam;Chompoonuch Khongla;Supaluk Sorapukdee
    • Animal Bioscience
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    • v.37 no.6
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    • pp.1096-1109
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    • 2024
  • Objective: This research aims to explore the nutritional and bioactive peptide properties of goat meat taken from various primal cuts, including the breast, shoulder, rib, loin, and leg, to produce these bioactive peptides during in vitro gastrointestinal (GI) digestion and absorption. Methods: The goat meat from various primal cuts was obtained from Boer goats with an average carcass weight of 30±2 kg. The meat was collected within 3 h after slaughter and was stored at -80℃ until analysis. A comprehensive assessment encompassed various aspects, including the chemical composition, cooking properties, in vitro GI digestion, bioactive characteristics, and the bioavailability of the resulting peptides. Results: The findings indicate that the loin muscles contain the highest protein and essential amino acid composition. When the meats were cooked at 70℃ for 30 min, they exhibited distinct protein compositions and quantities in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile, suggesting they served as different protein substrates during GI digestion. Subsequent in vitro simulated GI digestion revealed that the cooked shoulder and loin underwent the most significant hydrolysis during the intestinal phase, resulting in the strongest angiotensin-converting enzyme (ACE) and dipeptidyl peptidase-IV (DPP-IV) inhibition. Following in vitro GI peptide absorption using a Caco-2 cell monolayer, the GI peptide derived from the cooked loin demonstrated greater bioavailability and a higher degree of ACE and DPP-IV inhibition than the shoulder peptide. Conclusion: This study highlights the potential of goat meat, particularly cooked loin, as a functional meat source for protein, essential amino acids, and bioactive peptides during GI digestion and absorption. These peptides promise to play a role in preventing and treating metabolic diseases due to their dual inhibitory effects on ACE and DPP-IV.

Characteristics of 15 MV Photon Beam from a Varian Clinac 1800 Dual Energy Linear Accelerator (CLINAC 1800 선형가속기의 15 MV X-선의 특성)

  • Kim, Kye-Jun;Lee, Jong-Young;Park, Kyung-Ran
    • Radiation Oncology Journal
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    • v.9 no.1
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    • pp.131-141
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    • 1991
  • A comprehensive set of dosimetric measurements has been made on the Varian Clinac 1800 15 MV photon beam. Beam quality, percentage depth dose, dose in the build up region, output, symmetry and flatness, transmission through iead (Cerrobend), tray attenuation, isodose curves for the open and wedged fields were measured using 3 dimensional water phantom dosimetry system (including film densitometer system) and polystyrene phantoms. These dosimetric measurements sufficiently characterized the beam to permit clinical use. The depth dose characteristics of photon beam is $d_{max}$ of 3.0 cm and percentage depth dose of $76.8\%$ at 10 cm,100 cm source-surface distance, field size of $10\times10\;cm^2$ for 15 MV X-ray beam. The Output factors ranged 0.927 for $4\times4\;cm^2$ field to 1,087 for $35\times35\;cm^2$ field. The build-up level of maximum dose was at 3.0 cm and surface dose was approximately $15.5\%$ for a field size $10\times10\;cm^2$ The stability of output is $within\pm1\%$ and flatness and symmetry are $within\pm3\%$. The half value thickness (HVL) of lead is 13 mm, which corresponds to an attenuation coefficient of $0.053\;mm^{-1}$. These figures compare facorably with the manufacturesr`s specifications.

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Quantitative Assessments and Spatial Pattern Analyses of Weed Seed Banks of Arable Peat in Selangor, Malaysia (말레이지아 세랑고지역 부식질토양경지 매립잡초종자에 대한 정량생태분석)

  • Bakar, Baki Bin;Kwon, Yong-Woong;Yin, Fenny Wong Nyuk
    • Korean Journal of Weed Science
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    • v.17 no.3
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    • pp.269-280
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    • 1997
  • Collated data from the 1995-1996 field surveys of weed seeds buried in the plough layer of peat soil in Selangor district were analysed to assess species-dominance and spatial pattern of distribution of weed seeds based on selected quantitative indices and index of dispersion. Forty five species within 14 families were recorded of which 24 were broadleaves, 12 grasses and 9 sedges. They comprised ca. 53.2, 31.2 and 15.6%, respectively based on total population counts. Total seed population was ca. $8.14{\times}10^7$ seeds/ha within the fast 25cm soil depth. Wide variabilities in population counts were registered among species ranging from < $7.0{\times}10^4$ seeds/ha for Amaranthus gracilis to ca. $5.64{\times}10^6$ seeds/ha for Heteropogon contortus. Seeds of Cleome rutidesperma was the most abundant(ca. $2.347{\times}10^7$ seeds/ha). Difference in seed population counts may be attributed to inherent variation in fecundity, population fluxes, their spatial distribution patterns and the agronomic practices prevailing in the areas of survey. The profile distribution of soil seed banks was skewed within the first 0 - 10cm depth, comprising ca. 69% of the total seed counts. Seed counts in the 10 - 15, 15 - 20 and 20 - 25cm soil profiles were in the order of 17.9, 8.6 and 4.0% of the total populations, respectively. Weed seeds of all species displayed different degree of aggregated pattern of distribution with variance-to-mean ratios of > 1 and Lloyd's mean crowding($m^*$) values from 1.244 for Cyperus iria, Phyllanthus debilis, Phyllanthus urinaria, Scirpus grosses and urinaria lagopodiodes to 9607.7 for Cleome rutidosperma. Lloyd's patch indices(Ip) ranging from 5.1 for Aeschynomene indica to 188.5 for Bracharia reptans were registered. Differences in the VMR, $m^*$ and Ip values among species suggested inter-alia inherent variabilities in their disposal capacity from seed source and different agronomic practices prevailing in the areas surveyed.

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Skin Damage Sustained During Head-and-Neck and Shoulder Radiotherapy Due to the Curvature of Skin and the Use of Immobilization Mask (머리-목 그리고 어깨의 방사선 치료 시 피부곡면과 고정장치로 인한 피부손상연구)

  • Kim, Soo-Kil;Jeung, Tae-Sig;Lim, Sang-Wook;Park, Yeong-Mouk;Park, Dahl
    • Progress in Medical Physics
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    • v.21 no.1
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    • pp.86-92
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    • 2010
  • The purpose of this study was to measure curvature contour skin dose using radiochromic film and TLD for a conventional open field. We also attempted to quantify the degradation of skin sparing associated with use of immobilization devices for high energy photon beams and to calculate the skin dose with a help of Monte Carlo (MC) simulation. To simulate head-and-neck and shoulder treatment, a cylindrical solid water phantom 11 cm in diameter was irradiated with 6 MV x-rays using $40{\times}40\;cm^2$ field at 100 cm source axis distance (SAD) to the center of the phantom. Aquaplastic mesh mask was placed on the surface of the cylindrical phantom that mimicked relevant clinical situations. The skin dose profile was obtained by taking measurements from $0^{\circ}$ to $360^{\circ}$ around the circumference of the cylindrical phantom. The skin doses obtained from radiochromic film were found to be 47% of the maximum dose of $D_{max}$ at the $0^{\circ}$ beam entry position and 61% at the $90^{\circ}$ oblique beam position without the mask. Using the mask (1.5 mm), the skin dose received was 59% at $0^{\circ}$ incidence and 78% at $80^{\circ}$ incidence. Skin dose results were also gathered using thin thermoluminescent dosimeters (TLD). With the mask, the skin dose was 66% at $0^{\circ}$ incidence and 80% at $80^{\circ}$ incidence. This method with the mask revealed the similar pattern as film measurement. For the treatments of the head-and-neck and shoulder regions in which immobilization mask was used, skin doses at around tangential angle were nearly the same as the prescription dose. When a sloping skin contour is encountered, skin doses may be abated using thinner and more perforated immoblization devices which should still maintain immoblization.

A Study on the Optimum Design of Multiple Screw Type Dryer for Treatment of Sewage Sludge (하수슬러지 처리를 위한 다축 스크류 난류 접촉식 건조기의 최적 설계 연구)

  • Na, En-Soo;Shin, Sung-Soo;Shin, Mi-Soo;Jang, Dong-Soon
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.4
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    • pp.223-231
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    • 2012
  • The purpose of this study is to investigate basically the mechanism of heat transfer by the resolution of complex fluid flow inside a sophisticated designed screw dryer for the treatment of sewage sludge by using numerical analysis and experimental study. By doing this, the result was quite helpful to obtain the design criteria for enhancing drying efficiency, thereby achieving the optimal design of a multiple screw type dryer for treating inorganic and organic sludge wastes. One notable design feature of the dryer was to bypass a certain of fraction of the hot combustion gases into the bottom of the screw cylinder, by the fluid flow induction, across the delicately designed holes on the screw surface to agitate internally the sticky sludges. This offers many benefits not only in the enhancement of thermal efficiency even for the high viscosity material but also greater flexibility in the application of system design and operation. However, one careful precaution was made in operation in that when distributing the hot flue gas over the lump of sludge for internal agitation not to make any pore blocking and to avoid too much pressure drop caused by inertial resistance across the lump of sludge. The optimal retention time for rotating the screw at 1 rpm in order to treat 200 kg/hr of sewage sludge was determined empirically about 100 minutes. The corresponding optimal heat source was found to be 150,000 kcal/hr. A series of numerical calculation is performed to resolve flow characteristics in order to assist in the system design as function of important system and operational variables. The numerical calculation is successfully evaluated against experimental temperature profile and flow field characteristics. In general, the calculation results are physically reasonable and consistent in parametric study. In further studies, more quantitative data analyses such as pressure drop across the type and loading of drying sludge will be made for the system evaluation in experiment and calculation.

A marine deep-towed DC resistivity survey in a methane hydrate area, Japan Sea (동해의 메탄 하이드레이트 매장 지역에서의 해양 심부 견인 전기비저항 탐사)

  • Goto, Tada-Nori;Kasaya, Takafumi;Machiyama, Hideaki;Takagi, Ryo;Matsumoto, Ryo;Okuda, Yoshihisa;Satoh, Mikio;Watanabe, Toshiki;Seama, Nobukazu;Mikada, Hitoshi;Sanada, Yoshinori;Kinoshita, Masataka
    • Geophysics and Geophysical Exploration
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    • v.11 no.1
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    • pp.52-59
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    • 2008
  • We have developed a new deep-towed marine DC resistivity survey system. It was designed to detect the top boundary of the methane hydrate zone, which is not imaged well by seismic reflection surveys. Our system, with a transmitter and a 160-m-long tail with eight source electrodes and a receiver dipole, is towed from a research vessel near the seafloor. Numerical calculations show that our marine DC resistivity survey system can effectively image the top surface of the methane hydrate layer. A survey was carried out off Joetsu, in the Japan Sea, where outcrops of methane hydrate are observed. We successfully obtained DC resistivity data along a profile ${\sim}3.5\;km$ long, and detected relatively high apparent resistivity values. Particularly in areas with methane hydrate exposure, anomalously high apparent resistivity was observed, and we interpret these high apparent resistivities to be due to the methane hydrate zone below the seafloor. Marine DC resistivity surveys will be a new tool to image sub-seafloor structures within methane hydrate zones.

Establishment and Characterization of Bone Marrow Mesenchymal Stromal/Stem Cells (MSCs) Derived from ${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) Pig (${\alpha}$-1,3-Galactosyltransferase Knock Out(GalT KO) 돼지유래 골수 중간엽 줄기세포의 특성 규명)

  • Ock, Sun-A;Oh, Keon Bong;Hwang, Seongsoo;Im, Seoki;Kim, Youngim;Park, Jin-Ki
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.281-287
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    • 2013
  • A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

An Estimation of Concentration of Asian Dust (PM10) Using WRF-SMOKE-CMAQ (MADRID) During Springtime in the Korean Peninsula (WRF-SMOKE-CMAQ(MADRID)을 이용한 한반도 봄철 황사(PM10)의 농도 추정)

  • Moon, Yun-Seob;Lim, Yun-Kyu;Lee, Kang-Yeol
    • Journal of the Korean earth science society
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    • v.32 no.3
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    • pp.276-293
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    • 2011
  • In this study a modeling system consisting of Weather Research and Forecasting (WRF), Sparse Matrix Operator Kernel Emissions (SMOKE), the Community Multiscale Air Quality (CMAQ) model, and the CMAQ-Model of Aerosol Dynamics, Reaction, Ionization, and Dissolution (MADRID) model has been applied to estimate enhancements of $PM_{10}$ during Asian dust events in Korea. In particular, 5 experimental formulas were applied to the WRF-SMOKE-CMAQ (MADRID) model to estimate Asian dust emissions from source locations for major Asian dust events in China and Mongolia: the US Environmental Protection Agency (EPA) model, the Goddard Global Ozone Chemistry Aerosol Radiation and Transport (GOCART) model, and the Dust Entrainment and Deposition (DEAD) model, as well as formulas by Park and In (2003), and Wang et al. (2000). According to the weather map, backward trajectory and satellite image analyses, Asian dust is generated by a strong downwind associated with the upper trough from a stagnation wave due to development of the upper jet stream, and transport of Asian dust to Korea shows up behind a surface front related to the cut-off low (known as comma type cloud) in satellite images. In the WRF-SMOKE-CMAQ modeling to estimate the PM10 concentration, Wang et al.'s experimental formula was depicted well in the temporal and spatial distribution of Asian dusts, and the GOCART model was low in mean bias errors and root mean square errors. Also, in the vertical profile analysis of Asian dusts using Wang et al's experimental formula, strong Asian dust with a concentration of more than $800\;{\mu}g/m^3$ for the period of March 31 to April 1, 2007 was transported under the boundary layer (about 1 km high), and weak Asian dust with a concentration of less than $400\;{\mu}g/m^3$ for the period of 16-17 March 2009 was transported above the boundary layer (about 1-3 km high). Furthermore, the difference between the CMAQ model and the CMAQ-MADRID model for the period of March 31 to April 1, 2007, in terms of PM10 concentration, was seen to be large in the East Asia area: the CMAQ-MADRID model showed the concentration to be about $25\;{\mu}g/m^3$ higher than the CMAQ model. In addition, the $PM_{10}$ concentration removed by the cloud liquid phase mechanism within the CMAQ-MADRID model was shown in the maximum $15\;{\mu}g/m^3$ in the Eastern Asia area.