• Title/Summary/Keyword: experimental animal model

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CRISPR/Cas9-mediated generation of a Plac8 knockout mouse model

  • Lee, HyunJeong;Kim, Joo-Il;Park, Jin-Sung;Roh, Jae-il;Lee, Jaehoon;Kang, Byeong-Cheol;Lee, Han-Woong
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.279-287
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    • 2018
  • Placenta specific 8 (PLAC8, also known as ONZIN) is a multi-functional protein that is highly expressed in the intestine, lung, spleen, and innate immune cells, and is involved in various diseases, including cancers, obesity, and innate immune deficiency. Here, we generated a Plac8 knockout mouse using the CRISPR/Cas9 system. The Cas9 mRNA and two single guide RNAs targeting a region near the translation start codon at Plac8 exon 2 were microinjected into mouse zygotes. This successfully eliminated the conventional translation start site, as confirmed by Sanger sequencing and PCR genotyping analysis. Unlike the previous Plac8 deficient models displaying increased adipose tissue and body weights, our male Plac8 knockout mice showed rather lower body weight than sex-matched littermate controls, though the only difference between these two mouse models is genetic context. Differently from the previously constructed embryonic stem cell-derived Plac8 knockout mouse that contains a neomycin resistance cassette, this knockout mouse model is free from a negative selection marker or other external insertions, which will be useful in future studies aimed at elucidating the multi-functional and physiological roles of PLAC8 in various diseases, without interference from exogenous foreign DNA.

Ileal Amino Acid Digestibility of Broken Rice Fed to Postweaned Piglets with or without Multicarbohydrase and Phytase Supplementation

  • Dadalt, J.C.;Gallardo, C.;Polycarpo, G.V.;Budino, F.E.L.;Rogiewicz, A.;Berto, D.A.;Trindade Neto, M.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.10
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    • pp.1483-1489
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    • 2016
  • Most of amino acid (AA) digestibility values for feed ingredients are obtained using pigs cannulated in the distal ileum. The ileal-cannulated pig model uses pigs older than six weeks due to difficulties related to implanting the T-cannula in distal ileum of younger pigs and complications during the post-surgical recovery. However, to properly formulate the diet of weaned pigs, the nutritive value of feed ingredients should be determined with younger pigs. Thus, 25 weaned pigs were used to determine the apparent total tract digestibility (ATTD) of nutrients, energy, and apparent ileal digestibility (AID) and standardized ileal digestibility (SID) ileal AA digestibility of broken rice (BR), with or without multicarbohydrase (MC) and phytase (Phy) supplementation. Piglets were weaned at 23 d of age and individually housed in digestibility cages until 45 d of age. The trial consisted of 7 d of adaptation to the experimental diets and 3 d of excreta (feces and urine) collection. Ileal digesta was collected at slaughter (about 6 weeks of age). A completely randomized experimental design was used to determine the effects of MC and Phy. Reference diets (RD, 5% casein) was replaced by 30% of BR with or without MC, Phy, or MC+Phy. The RD was used to quantify endogenous AA losses. BR with Phy supplied had increased the ATTD of dry matter (p<0.05) and SID of histidine (p = 0.05), arginine, leucine, lysine, valine, alanine, and proline (p<0.05). BR with MC had been increased digestible energy and protein and SID for histidine (p<0.05). There was no interaction between Phy and MC on the BR nutrient digestibilities. Standardized amino acid digestibilities of BR, without enzymes, were lower than those values reported in the literature. The MC and Phy improved the digestibility of some nutrients and energy of BR in post-weaned piglet diets.

An orthotopic nude mouse model of tongue carcinoma (구강암 세포주를 이종이식한 설암의 동소위 누드마우스 모델)

  • Chung, Jae-Seung;Kim, So-Mi;Hwang, Young-Sun;Zhang, Xianlan;Cha, In-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.6
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    • pp.490-495
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    • 2011
  • Introduction: Development of carcinoma on oral tongue may cause bilateral cervical lymph node metastasis, rapid invasion and growth of the cancer cells due to rich blood supply in muscle tissues. It is not only difficult to develop an animal experimental model, but also to proceed follow-up research after the development of such model as the induction of cancer lead to difficulty in taking nutrition for the experimental animals that often causes early death. Materials and Methods: IIn this study, author have transplanted YD-$10B_{mod}$ cells into nude mouse oral tongues with different cells number ($5{\times}10^4$, $5{\times}10^5$, $5{\times}10^6$ cells/mouse) and observed the development aspect of oral tongue cancers. Results: The cancer developed from orthotopic transplantation of YD-$10B_{mod}$ cells into nude mouse oral tongue show invasion and central necrosis of the tumor, similar to the cancers developed human oral tongue cancer. The difference in tumor size and the time of central necrosis development depending on the number of transplanted tumor cells shows the feasibility of extending the survival period of the nude mouse by limiting the transplanted tumor cells to < $5{\times}10^4$ cells/mouse or under per nude mouse. Conclusion: This nude mouse model could be used effectively in developing effective chemotheray agent and establishing an animal experimental model that can be used to study the mechanism of cervical lymph node metastasis of the oral tongue cancer.

Evaluating sulfoxaflor residues in pig tissues using animal modeling

  • Hyun-Woo, Cho;Kangmin, Seo;Jin Young, Jeong;Ju Lan, Chun;Ki Hyun, Kim
    • Journal of Animal Science and Technology
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    • v.64 no.5
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    • pp.911-921
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    • 2022
  • Maximum residue limits (MRL) for pesticides in feed have been set to protect public health and produce safe livestock products. In vivo experiments to establish MRL are essential, as livestock are commonly used to obtain reliable in vivo quantitative information. Here, we aimed to evaluate whether small laboratory animals can replace or reduce monogastric livestock in experiments to quantify pesticide residues in vivo after oral consumption through feed. First, 24 pigs and rats were randomly assigned to four groups and fed 0, 3, 9, or 30 mg/kg of sulfoxaflor. After four weeks, serum, muscle, fat, liver, kidney, and small intestine samples were collected, and sulfoxaflor residues were analyzed using liquid chromatography - tandem mass spectrometry. Sulfoxaflor residues in pig tissues were significantly correlated with those in rat tissues. Model equations were formulated based on the residual sulfoxaflor amount in pig and rat tissues. The calculated and measured sulfoxaflor residues in pigs and rats showed more than 90% similarity. Sulfoxaflor did not affect body weight gain, feed intake, or the feed conversion ratio. Therefore, we concluded that pesticide residue quantification in vivo to establish MRL could be performed using small laboratory animals instead of livestock animals. This would contribute to obtaining in vivo pesticide residue information and reducing large-scale livestock animal experiments.

Detection of Imprinted Quantitative Trait Loci (QTL) for Growth Traits in Pigs

  • Lee, H.K.;Lee, S.S.;Kim, T.H.;Jeon, G.J.;Jung, H.W.;Shin, Y.S.;Han, J.Y.;Choi, B.H.;Cheong, I.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.8
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    • pp.1087-1092
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    • 2003
  • As an experimental reference population, crosses between Korean native pig and Landraces were established and information on growth traits was recorded. Animals were genotyped for 24 microsatellite markers covering chromosomes 2, 6, and 7 for partial-genome scan to identify chromosomal regions that have effects on growth traits. quantitative trait loci (QTL) effects were estimated using interval mapping by the regression method under the line cross models with a test for imprinting effects. For test of presence of QTL, chromosome-wide and single position significance thresholds were estimated by permutation test and normal significance threshold for the imprinting test were derived. For tests against the Mendelian model, additive and dominance coefficients were permuted within individuals. Thresholds (5% chromosome-wide) against the no-QTL model for the analyzed traits ranged from 4.57 to 4.99 for the Mendelian model and from 4.14 to 4.67 for the imprinting model, respectively. Partial-genome scan revealed significant evidence for 4 QTL affecting growth traits, and 2 out of the 4 QTLs were imprinted. This study demonstrated that testing for imprinting should become a standard procedure to unravel the genetic control of multi-factorial traits. The models and tests developed in this study allowed the detection and evaluation of imprinted QTL.

Review of Experimental Researches on Bee Venom Pharmacopuncture Therapy for Chemotherapy-induced Peripheral Neuropathy (항암화학요법 유발 말초신경병증에 대한 봉독 약침 요법의 효과 및 기전에 대한 실험연구 고찰)

  • Kwon, Bo In;Woo, Yeonju;Kim, Joo-Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.35 no.1
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    • pp.1-7
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    • 2021
  • Chemotherapy-induced peripheral neuropathy (CIPN) is one of the most common dose-limiting side effects of neurotoxic chemotherapeutic agents that lead to decreased quality of life and dose reduction, delay or even cessation of treatment. The purpose of this systematic review is to evaluate the effect and the underlying mechanisms of bee venom (BV) pharmacopuncture therapy for CIPN in animal models. We searched for the available experimental literature using BV for CIPN through the Pubmed databases. Ten experimental studies were finally included in this review. In the oxaliplatin or paclitaxel-induced CIPN animal model, BV significantly relieved pain caused both mechanical and cold stimulation. It was suggested that the effect of BV is mediated by the stimulation effect of spinal α1- and α2-adrenergic receptors as a potential mechanism. In the future, more experimental studies are needed.

An assessment of responses to egg production and liver health of Japanese quails subjected to different levels of metabolizable energy

  • Diana Maryuri Correa, Castiblanco;Michele Bernardino, de Lima;Silvana Martinez Baraldi, Artoni;Erikson Kadoshe de Morais, Raimundo;Daniel Silva, Santos;Lizia Cordeiro, de Carvalho;Edney Pereira, da Silva
    • Animal Bioscience
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    • v.36 no.1
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    • pp.98-107
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    • 2023
  • Objective: Current quail production is configured as an economic activity in scale. Advancements in quail nutrition have been limited to areas such as breeding and, automation of facilities and ambience. The objective of this study was to evaluate the performance responses, liver and oviduct morphometry, and liver histology of Japanese laying quails subjected to different levels of nitrogen-corrected apparent metabolizable energy (MEn). Methods: A completely random design was used that consisted of nine levels of MEn, six replicates, and five hens per cage with a total of 270 quails. The experimental period lasted for 10 weeks. The variables of performance were subjected to analysis of variance and then regression analysis using the broken-line model. The morphometric and histological variables were subjected to multivariate exploratory techniques. Results: The MEn levels influenced the responses to zootechnical performance. The broken-line model estimated the maximum responses for feed intake, egg production, egg weight, and egg mass as 3,040, 2,820, 1,802, and 2,960 kcal of MEn per kg of diet, respectively. Multivariate analysis revealed that the occurrence of hepatic steatosis and increased levels of Kupffer cells were not related to MEn levels. Conclusion: The level of 2,960 kcal/kg of MEn meets performance variable requirements without compromising hepatic physiology.

The Effect of Herbal Medicine on Rheumatoid Arthritis Animal Model: A systematic Review of Animal Experiment Studies Published in Korea and Overseas (한약의 류마티스 관절염 동물 모델에 대한 치료효과: 국내 및 국외 논문에 대한 체계적 문헌 고찰)

  • Oh, Seung-Joon;Ha, Hyun Ju;Gu, Ji-Hyang;Park, Yang-Chun;Oh, Min-Seok;Jung, In-Chul;Lee, Eun-Jung
    • Journal of Korean Medicine Rehabilitation
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    • v.30 no.3
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    • pp.103-116
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    • 2020
  • Objectives This study was designed to review the effect of herbal medicine on animal model with rheumatoid arthritis. Methods We reviewed studies published both in Korea and overseas, all published after 2008. We investigated 8 databases, 6 Korean databases and 2 of abroad. Searching was conducted through flowchart and 24 studies met our guideline. Datas were extrated and organized as animal model characteristics, experimental period, study design, herbal medicine used as intervention and rheumatoid arthritis indicator. Results 336 articles were primarily selected. After screening, 24 articles were left in the end. All 24 studies showed that the herbal medicine used in the animal experiment was effective on treating rheumatoid arthris, proven by macroscopic, histological, hematological and osteological indicators. Conclusions The result showed that herbal medicine is effective in treating rheumatoid arthritis through various pathways. However, study design and intervention were all different from each other and we could not specify the certain compound, material or substance of efficacy. Further study should be conducted to figure out the effectiveness of herbal medicine on rheumatoid arthritis.

CRISPR/Cas9-mediated knockout of Rag-2 causes systemic lymphopenia with hypoplastic lymphoid organs in FVB mice

  • Kim, Joo-Il;Park, Jin-Sung;Kim, Hanna;Ryu, Soo-Kyung;Kwak, Jina;Kwon, Euna;Yun, Jun-Won;Nam, Ki-Taek;Lee, Han-Woong;Kang, Byeong-Cheol
    • Laboraroty Animal Research
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    • v.34 no.4
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    • pp.166-175
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    • 2018
  • Recombination activating gene-2 (RAG-2) plays a crucial role in the development of lymphocytes by mediating recombination of T cell receptors and immunoglobulins, and loss of RAG-2 causes severe combined immunodeficiency (SCID) in humans. Rag-2 knockout mice created using homologous recombination in ES cells have served as a valuable immunodeficient platform, but concerns have persisted on the specificity of Rag-2-related phenotypes in these animals due to the limitations associated with the genome engineering method used. To precisely investigate the function of Rag-2, we recently established a new Rag-2 knockout FVB mouse line ($Rag-2^{-/-}$) manifesting lymphopenia by employing a CRISPR/Cas9 system at Center for Mouse Models of Human Disease. In this study, we further characterized their phenotypes focusing on histopathological analysis of lymphoid organs. $Rag-2^{-/-}$ mice showed no abnormality in development compared to their WT littermates for 26 weeks. At necropsy, gross examination revealed significantly smaller spleens and thymuses in $Rag-2^{-/-}$ mice, while histopathological investigation revealed hypoplastic white pulps with intact red pulps in the spleen, severe atrophy of the thymic cortex and disappearance of follicles in lymph nodes. However, no perceivable change was observed in the bone marrow. Moreover, our analyses showed a specific reduction of lymphocytes with a complete loss of mature T cells and B cells in the lymphoid organs, while natural killer cells and splenic megakaryocytes were increased in $Rag-2^{-/-}$ mice. These findings indicate that our $Rag-2^{-/-}$ mice show systemic lymphopenia with the relevant histopathological changes in the lymphoid organs, suggesting them as an improved Rag-2-related immunodeficient model.