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Exosomes from Tension Force-Applied Periodontal Ligament Cells Promote Mesenchymal Stem Cell Recruitment by Altering microRNA Profiles

  • Maolin Chang;Qianrou Chen;Beike Wang;Zhen Zhang;Guangli Han
    • International Journal of Stem Cells
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    • v.16 no.2
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    • pp.202-214
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    • 2023
  • Background and Objectives: To investigate the role of exosomes from periodontal ligament cells (PDLCs) in bone marrow mesenchymal stem cell (BMSC) migration. Methods and Results: Human PDLCs were applied cyclic tension stretching. Exosomes were extracted from cultured PDLCs by ultracentrifugation, then characterized for their size, morphology and protein markers by NTA, TEM and western blotting. The process that PKH26-labeled exosomes taken up by BMSCs was assessed by confocal microscope. BMSC migration was examined by Transwell assay. Exosomes derived from PDLCs were identified. Cyclic tension stretch application on PDLCs can enhance the migration ability of BMSCs through exosomes. The exosomal miRNA expression profiles of unstretched and stretched PDLCs were tested by miRNA microarray. Four miRNAs (miR-4633-5p, miR-30c-5p, miR-371a-3p and let-7b-3p) were upregulated and six (miR-4689, miR-8485, miR-4655-3p, miR-4672, miR-3180-5p and miR-4476) were downregulated in the exosomes after stretching. Sixteen hub proteins were found in the miRNA-mRNA network. Gene Ontology and KEGG pathway analyses demonstrated that the target genes of differentially expressed exosomal miRNAs closely related to the PI3K pathway and vesicle transmission. Conclusions: The exosomes derived from cyclic tension-stretched PDLCs can promote the migration of BMSCs. Alternation of microRNA profiles provides a basis for further research on the regulatory function of the exosomal miRNAs of PDLCs during orthodontic tooth movement.

CysLT receptor-mediated NOX2 activation is required for IL-8 production in HMC-1 cells induced by Trichomonas vaginalis-derived secretory products

  • Young Ah Lee;Myeong Heon Shin
    • Parasites, Hosts and Diseases
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    • v.62 no.3
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    • pp.270-280
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    • 2024
  • Trichomoniasis is caused by a sexually transmitted flagellate protozoan parasite Trichomonas vaginalis. T. vaginalis-derived secretory products (TvSP) contain lipid mediators such as leukotriene B4 (LTB4) and various cysteinyl leukotrienes (CysLTs) which included LTC4, LTD4, and LTE4. However, the signaling mechanisms by which T. vaginalis-induced CysLTs stimulate interleukin (IL)-8 production in human mast cells remain unclear. In this study, we investigated these mechanisms in human mast cells (HMC-1). Stimulation with TvSP resulted in increased intracellular reactive oxygen species (ROS) generation and NADPH oxidase 2 (NOX2) activation compared to unstimulated cells. Pre-treatment with NOX2 inhibitors such as diphenyleneiodonium chloride (DPI) or apocynin significantly reduced ROS production in TvSP-stimulated HMC-1 cells. Additionally, TvSP stimulation increased NOX2 protein expression and the translocation of p47phox from the cytosol to the membrane. Pretreatment of HMC-1 cells with PI3K or PKC inhibitors reduced TvSP-induced p47phox translocation and ROS generation. Furthermore, NOX2 inhibitors or NOX2 siRNA prevented CREB phosphorylation and IL-8 gene expression or protein secretion induced by TvSP. Pretreatment with a CysLTR antagonist significantly inhibited TvSP-induced ROS production, CREB phosphorylation, and IL-8 production. These results indicate that CysLT-mediated activation of NOX2 plays a crucial role in ROS-dependent IL-8 production in human mast cells stimulated by T. vaginalis-secreted CysLTs. These findings enhance our understanding of the inflammatory response in trichomoniasis and may inform the development of targeted therapies to mitigate this response.

Experimental Pathogenesis of Pullorum Disease with the Local Isolate of Salmonella enterica serovar. enterica subspecies Pullorum in Pullets in Bangladesh

  • Haider, M.G.;Chowdhury, E.H.;Khan, M.A.H.N.A.;Hossain, M.T.;Rahman, M.S.;Song, H.J.;Hossain, M.M.
    • Korean Journal of Poultry Science
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    • v.35 no.4
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    • pp.341-350
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    • 2009
  • The research work was carried out to study the pathogenesis covering the clinical signs, gross and histopathological lesions in different organs, and reisolation and identification of the organisms after experimental infection with the local isolate of Salmonella enterica serovar. enterica subspecies (S.) Pullorum at different time interval of the experiment during the period February 2006 to December 2006. One hundred pullets (seronegative to S. Pullorum of 12 weeks age were purchased and divided into 5 (A, B, C, D and E) groups and each group consisted of 20 birds. Four groups (A, B, C and D) were infected orally with a dose of $10^6\;CFU$, $10^7\;CFU$, $2{\times}10^7\;CFU$, $10^8\;CFU$ of S. Pullorum, respectively, and one group (E) was treated as uninfected control. The used methods were necropsy and histopathology, culture of bacteria, staining and biochemical test of Salmonella. Five birds from each group were randomly selected and sacrificed $1^{st}$ week, $2^{nd}$, $3^{rd}$ and $4^{th}$ weeks of post infection (PI). From all the groups, the bacteriological samples (crop, liver, lung, heart, spleen, bile duodenum, ceca and blood) were collected with pre enriched in buffered peptone water in sterile poly bags. Liver, lungs, heart, spleen, intestine, etc. were collected in 10% buffered-formalin for histopathological examination. No clinical signs, gross and histopathological lesions were found in control group and no S. Pullorum was reisolated. Clinical sign of experimentally infected with S. Pullorum in pullets were loss of appetite (100%), slight depression (75%), ruffled feathers (85%), diarrhea (60%) and loss of weight (100%) in chickens. The feed intake and body weight at different weeks after PI differed significantly (p<0.01) among the groups. Grossly, the highest recorded lesion was button-like ulcer in the ceca (80%) and the lowest was white nodules in lungs (1.25%). S. Pullorum were reisolated from crop (91.25%), liver (91.25%), lung (83.75%), heart (71.25%), spleen (87.75%), bile (33.25%), duodenum (92.50%), ceca (97.50%) and from different group of infection (61.25%). The highest microscopic findings were intestinal and cecal mucosa and submucosa exhibited infiltration of mononuclear cells and congestion (96.25%), and the lowest finding was nodule formation in the lungs (3.75%). The pattern of the disease production by local isolate of S. Pullorum in Bangladesh is almost similar with other isolates in different countries.

A New Early-Heading and High-Yielding Winter Oat Cultivar for Whole Crop Forage, 'Okhan' (조숙 다수성 추파 총체사료용 귀리 신품종 '옥한')

  • Han, Ouk-Kyu;Park, Tae-Il;Park, Hyung-Ho;Song, Tae-Hwa;Kim, Kee-Jong;Park, Nam-Geon;Ju, Jung-Il;Jang, Young-Jik;Hwang, Jong-Jin;Kwon, Young-Up
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.2
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    • pp.87-93
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    • 2013
  • 'Okhan' (Avena sativa L.), an oat cultivar for winter use, was developed by the breeding team at the Department of Rice and Winter Cereal Crop, NICS, RDA in 2011. It was derived from an original cross between 'Early80', exhibiting early heading, and 'Maine PI-590' (CI 7518), exhibiting large-size grain, in 1995. Subsequent generations as well as cross-bred cultivars were handled in bulk, and pedigree selection programs took place at Suwon and Yeoncheon, respectively. A promising line, 'SO95027-B-45-16-10-6-2-Y7-10', was selected in 2004, and was designated 'Gwiri74' after being selected from a yield trial for three years from 2005 to 2008. The breeding line 'Gwiri74' was subsequently evaluated for earliness of heading and forage yield in four different locations, Yesan, Iksan, Kimjae, and Jeju, from 2009 to 2011, and was finally named as 'Okhan'. Over 3 years, the heading date of 'Okhan' was about 6 days earlier than that of the check cultivar 'Samhan', and their average forage dry matter yield harvested at the milk-ripe stage was 15.0 ton $ha^{-1}$, compared with 14.1 ton $ha^{-1}$ of check cultivar. Cultivar 'Okhan' was lower than the check cultivar 'Samhan' in terms of the protein content (9.2% and 9.9%, respectively) and total digestible nutrients (58.5%, and 59.3%, respectively), while the TDN yield per ha was more than the check (8.70 and 8.36 kg, respectively). Fall sowing cropping of 'Okhan' is recommended only in areas where average daily minimum mean temperatures in January are higher than $-7^{\circ}C$, and it should not be cultivated in mountain areas, where frost damage is likely to occur.

Inheritance of Kunitz Trypsin Inhibitor and P34 Protein in Soybean Seed (콩 종자에서 쿠니츠트립신인히비터와 P34 단백질의 유전)

  • Han, Eun-Hui;Sung, Mi-Kyung;Baek, Woon-Jang;Shim, Sang-In;Kim, Min-Chul;Chung, Jong-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.1
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    • pp.78-82
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    • 2012
  • Soybean [$Glycine$ $max$ (L.) Merr.] protein is a high quality source for food and feed. But, antinutritional factors in the raw mature soybean are exist. Kunitz trypsin inhibitor (KTI) protein is a main antinutritional factor in soybean seed. Also, P34 protein, referred as $Gly$ $m$ Bd 30K, has been identified as a predominant immunodominant allergen. Genetic relationship between KTI protein and P34 protein could be useful in soybean breeding program for the genetic elimination or reduction of these factors. The objective of this study was to determine the independent inheritance or linkage between KTI protein and P34 protein in soybean seed. A total of 479 $F_2$ seeds were obtained from the cross of 07B1 and PI567476 parents. KTI protein and relative amount of P34 protein were analysed from $F_2$ seeds harvested from the F1 plants by using SDS-PAGE and Western blot analysis. The segregation ratios of 3 : 1 for KTI protein (353 KTI protein present : 126 KTI protein absent) and relative amount of P34 protein (363 normal amount of P34 protein : 116 low amount of P34 protein). The segregation ratio of 3 : 1 suggested that KTI protein and relative amount of P34 protein in mature soybean seed were controlled by a single major gene. The segregation ratios of 9 : 3 : 3 : 1 (266 KTI protein present, normal amount of P34 protein: 88 KTI protein present, low amount of P34 protein: 102 KTI protein absent, normal amount of P34 protein: 23 KTI protein absent, low amount of P34 protein) and Chi-square value (${\chi}^2$=3.31, P=0.346) were observed in $F_2$ seeds. This data showed that KTI protein was inherited independently with relative amount of P34 protein in soybean. These results will be helpful in breeding program for selecting the line with lacking KTI protein and reduced amount of P34 protein in soybean.

A study on the effect of microgroove-fibronectin complex titanium plate on the expression of various cell behavior-related genes in human gingival fibroblasts (인간치은섬유아세포의 다양한 세포행동 관련 유전자발현에 마이크로그루브-파이브로넥틴 복합 티타늄표면이 미치는 영향에 대한 연구)

  • Hwang, Yu Jeong;Lee, Won Joong;Leesungbok, Richard;Lee, Suk Won
    • Journal of Dental Rehabilitation and Applied Science
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    • v.38 no.3
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    • pp.150-161
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    • 2022
  • Purpose: To determine the effects of the microgroove-fibronectin complex surface on the expression of various genes related to cellular activity in human gingival fibroblasts. Materials and Methods: Smooth titanium specimens (NE0), acid-treated titanium specimens (E0), microgroove and acid-treated titanium specimens (E60/10), fibronectin-fixed smooth titanium specimens (NE0FN), acid-treated and fibronectin-immobilized titanium specimens (E0FN), and microgroove and acid-treated titanium specimens immobilized with fibronectin (E60/10FN) were prepared. Real-time polymerase chain reaction experiments were conducted on 44 genes related to cell behavior of human gingival fibroblasts. Results: Adhesion and proliferation of human gingival fibroblast on microgroove-fibronectin complex titanium were activated through four types of signaling pathway. Integrin α5, Integrin β1, Integrin β3, Talin-2, which belong to the focal adhesion pathway, AKT1, AKT2, NF-κB, which belong to the PI3K-AKT signaling pathway, MEK2, ERK1, ERK2, which belong to the MAPK signaling pathway, and Cyclin D1, CDK4, CDK6 genes belonging to the cell cycle signaling pathway were upregulated on the microgroove-fibronectin complex titanium surface (E60/10FN). Conclusion: The microgroove-fibronectin complex titanium surface can up-regulate various genes involved in cell behavior.

Molecular-epidemiologic study on outbreak of colonization by extended spectrum β-lactamase producing Klebsiella pneumoniae in neonatal intensive care unit (신생아 중환자실에서 extended spectrum β-lactamase를 생성하는 Klebsiella pneumoniae 집단 보균 발생의 분자 역학적 조사 및 추적관찰)

  • Jun, Nu-Lee;Kim, Mi-Na;Jeong, Jae-Sim;Kim, Yang-Soo;Kim, Ellen Ai-Rhan;Kim, Ki-Soo;Pi, Soo-Young
    • Clinical and Experimental Pediatrics
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    • v.49 no.2
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    • pp.150-156
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    • 2006
  • Purpose : The aims of this study included assessment of molecular-epidemiologic features during an outbreak of colonization of extended spectrum ${\beta}$-lactamase producing Klebsiella pneumoniae(ESBL-KPN) and re-evaluation of their colonized status one year later. Methods : Rectal swab cultures for ESBL-KPN from all hospitalized infants and newly admitted infants were obtained during the outbreak of colonization from July to December, 2000. The pattern of XbaI-digested chromosomal DNA of isolates were analyzed by pulsed-field gel electrophoresis. Weekly rectal swab cultures were obtained during the outbreak until patients were either discharged or decolonized. Patients discharged after being colonized had follow up stool cultures a year later. Results : A total of 80 patients(28.5 percent) were colonized. Of those, 53 whose pulsed-field gel electrophoresis(PFGE) was possible only once, were ESBL-KPN grouped into six cluster clones and 10 single clones : 28 patients(52.8 percent) were colonized with type A, the most common clone, followed by type B in 11 patients(20.8 percent). Of those 12 patients in whom serial PFGE was done more than twice, type A was predominant. Narrowed-down in strains occurred from types A, B, C, D and three single clones at initiation of the study into types A and type B after three months of strict infection control. Among 75 patients(93.7 percent) who were sent home after being colonized, 30 patients were re-called for stool cultures a year later : All of them were decolonized. Conclusion : This study demonstrates the importance of infection control as the diversity of ESBL-KPN strains could be narrowed into fewer strains. Colonization of ESBL-KPN could be reversed upon return to the community.

Photochemical Response Analysis on Drought Stress for Red Pepper (Capsiumannuum L.)

  • Yoo, Sung-Yung;Lee, Yong-Ho;Park, So-Hyun;Choi, Kyong-Mi;Park, June-Young;Kim, A-Ram;Hwang, Su-Min;Lee, Min-Ju;Ko, Tae-Seok;Kim, Tae-Wan
    • Korean Journal of Soil Science and Fertilizer
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    • v.46 no.6
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    • pp.659-664
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    • 2013
  • The aim of this study is to determine the drought stress index through photochemical analysis in red pepper (Capsiumannuum L.). The photochemical interpretation was performed in the basis of the relation between Kautsky effect and Photosystem II (PSII) following the measurement of chlorophyll, pheophytin contents, and $CO_2$ assimilation in drought stressed 5-week-old red pepper plants. The $CO_2$ assimilation rate was severely lowered with almost 77% reduction of chlorophyll and pheophytin contents at four days after non-irrigation. It was clearly observed that the chlorophyll fluorescence intensity rose from a minimum level (the O level), in less than one second, to a maximum level (the P-level) via two intermediate steps labeled J and I (OJIP process). Drought factor index (DFI) was also calculated using measured OJIP parameters. The DFI was -0.22, meaning not only the initial inhibition of PSII but also sequential inhibition of PSI. In real, most of all photochemical parameters such as quantum yield of the electron transport flux from Quinone A ($Q_A$) to Quinone B ($Q_B$), quantum yield of the electron transport flux until the PSI electron acceptors, quantum yield of the electron transport flux until the PSI electron acceptors, average absorbed photon flux per PSII reaction center, and electron transport flux until PSI acceptors per cross section were profoundly reduced except number of QA reducing reaction centers (RCs) per PSII antenna chlorophyll (RC/ABS). It was illuminated that at least 6 parameters related with quantum yield/efficiency and specific energy fluxes (per active PSII RC) could be applied to be used as the drought stress index. Furthermore, in the combination of parameters, driving forces (DF) for photochemical activity could be deduced from the performance index (PI) for energy conservation from photons absorbed by PSII antenna until the reduction of PSI acceptors. In conclusion, photochemical responses and their related parameters can be used as physiological DFI.

Characterization of Phosphoinositide-3-kinase, Class 3 (PIK3C3) Gene and Association Tests with Quantitative Traits in Pigs

  • Kim, J.H.;Choi, B.H.;Lim, H.T.;Park, E.W.;Lee, S.H.;Seo, B.Y.;Cho, I.C.;Lee, J.G.;Oh, S.J.;Jeon, J.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.12
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    • pp.1701-1707
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    • 2005
  • This study deals with the characterization of porcine PIK3C3 and association tests with quantitative traits. PIK3C3 belongs to the class 3 PI3Ks that participate in the regulation of hepatic glucose output, glycogen synthase, and antilipolysis in typical insulin target cells such as those in the such as liver, muscle system, and fat. On the analysis of full-length mRNA sequence, the length of the PIK3C3 CDS was recorded as 2,664 bps. As well, nucleotide and amino acid identities between human and pig subjects were 92% and 99%, respectively. Five SNPs were detected over 5 exons. We performed genotyping by using a SNP C2604T on exon24 for 145 F$_2$ animals (from a cross between Korean native boars and Landrace sows) by PCR-RFLP analysis with Hpy8I used to investigate the relationship between growth and fat depot traits. In the total association analysis, which doesn' consider transmission disequilibrium, the SNP showed a significant effect (p<0.05) on body weight and carcass fat at 30 weeks of age as well as a highly significant effect (p<0.01) on back fat. In an additional sib-pair analysis, C allele still showed positive and significant effects (p<0.05) on back fat thickness and carcass fat. Moreover, the effects of C allele on the means of within-family components for carcass fat and back fat were estimated as 2.76 kg and 5.07 mm, respectively. As a result, the SNP of porcine PIK3C3 discovered in this study could be utilized as a possible genetic marker for the selection of pigs that possess low levels of back fat and carcass fat at the slaughter weight.

Insecticidal Activity of N'-phenvl-N-Methylformamidine Analogues against Two Spotted Spider Mite (Tetranychus urticae) and Design of New Potent Compounds (두 점박이 응애(Tetranychus urticae)에 대한 N'-phenyl-N-methylformamidine 유도체의 살충활성과 새로운 고활성 화합물들의 설계)

  • Lee, Jae-Whang;Choi, Won-Seok;Lee, Dong-Guk;Chung, Kun-Hoe;Ko, Young-Kwan;Kim, Tae-Joon;Sung, Nack-Do
    • The Korean Journal of Pesticide Science
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    • v.14 no.3
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    • pp.191-198
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    • 2010
  • To predict and design of new potent insecticidal compounds, the two dimensional quantitative structure-activity relationships (2D-QSARs) and molecular hologram quantitative structure-activity relationships (HQSARs) between the various physicochemical parameters as descripters of N'-phenyl-N-methylformamidine analogues (1-22) and their insecticidal activity against the two spotted spider mite (Tetranychus urticae) were discussed quantitatively. From 2D-QSAR models (1 & 3), the width ($B_2$) of $R_3$-group as sterically factor and optimal total dipole moment (TDM=2.025D) of $R_4$-group were mainly influenced to increase the activity. Therefore, the activities were depend upon the $R_3$- and $R_4$-groups. Particularly, it is predicted that the activity of newly designed potent compound (PI; $EC_{50}$=0.516 ppm) by 2D-QSAR models (3) and HQSAR model F2 was about 34.3 fold higher than that of the commercialized insecticide, Amitraz ($EC_{50}$=17.7 ppm).