• Title/Summary/Keyword: direct differentiation

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Ribose-induced Maillard Reaction as an Analytical Method for Detection of Adulteration and Differentiation of Chilled and Frozen-thawed Minced Veal

  • Akbarabadi, Masoumeh;Mohsenzadeh, Mohammad;Housaindokht, Mohammad-Reza
    • Food Science of Animal Resources
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    • v.40 no.3
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    • pp.350-361
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    • 2020
  • Quality control of meat products is one of the main concerns of consumers, governmental control authorities, and retailers. The purpose of this study was to employ ribose-induced Maillard reaction in detection of meat adulteration and differentiation of fresh-chilled from frozen-thawed minced veal. The browning intensity was assessed through measuring the absorbance at 420 nm with a spectrophotometer as well as the direct analysis of the color and pH. The results showed that CIE b*, CIE a*, and A420* values in the extract of fresh-chilled veal were significantly (p<0.05) higher than frozen-thawed samples. The extract of frozen meat samples stored at -18℃ became significantly darker and more yellowish compared to -4℃. The results showed that the A420* value in the frozen-thawed veal stored at -4℃ and -18℃ was reduced by approximately 17.22±3.53% and 11.68±2.49%, respectively, compared with fresh-chilled veal. The findings also showed that the storage temperature of minced veal and the heating time in this reaction had a significant effect on all tested variables (p<0.0001). The proposed method can be considered as an easy, quick, and inexpensive test for differentiating between the fresh-chilled and frozen-thawed minced veal.

No Relevance of NF-${\kappa}B$ in the Transcriptional Regulation of Human Nanog Gene in Embryonic Carcinoma Cells

  • Seok, Hyun-Jeong;Kim, Young-Eun;Park, Jeong-A;Lee, Young-Hee
    • Development and Reproduction
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    • v.15 no.1
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    • pp.25-30
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    • 2011
  • Embryonic stem (ES) cells can self-renew maintaining the undifferentiated state. Self renewal requires many factors such as Oct4, Sox2, FoxD3, and Nanog. NF-${\kappa}B$ is a transcription factor involved in many biological activities. Expression and activity of NF-${\kappa}B$ increase upon differentiation of ES cells. Reportedly, Nanog protein directly binds to NF-${\kappa}B$ protein and inhibits its activity in ES cells. Here, we found a potential binding site of NF-${\kappa}B$ in the human Nanog promoter and postulated that NF-${\kappa}B$ protein may regulate expression of the Nanog gene. We used human embryonic carcinoma (EC) cells as a model system of ES cells and confirmed decrease of Nanog and increase of NF-${\kappa}B$ upon differentiation induced by retinoic acid. Although deletion analysis on the DNA fragment including NF-${\kappa}B$ binding site suggested involvement of NF-${\kappa}B$ in the negative regulation of the promoter, site-directed mutation of NF-${\kappa}B$ binding site had no effect on the Nanog promoter activity. Furthermore, no direct association of NF-${\kappa}B$ with the Nanog promoter was detected during differentiation. Therefore, we conclude that NF-${\kappa}B$ protein may not be involved in transcriptional regulation of Nanog gene expression in EC cells and possibly in ES cells.

Regulation of Pluripotency-related Genes and Differentiation in Mouse Embryonic Stem Cells by Direct Delivery of Cell-penetrating Peptide-conjugated CARM1 Recombinant Protein

  • Choi, Sara;Jo, Junghyun;Seol, Dong-Won;Cha, Soo Kyung;Lee, Jeoung Eun;Lee, Dong Ryul
    • Development and Reproduction
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    • v.17 no.1
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    • pp.9-16
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    • 2013
  • Coactivator-associated arginine methyltransferase 1 (CARM1) is included in the protein arginine methyltransferase (PRMT) family, which methylates histone arginine residues through posttranslational modification. It has been proposed that CARM1 may up-regulate the expression of pluripotency-related genes through the alteration of the chromatin structure. Mouse embryonic stem cells (mESCs) are pluripotent and have the ability to self-renew. The cells are mainly used to study the genetic function of novel genes, because the cells facilitate the transmission of the manipulated genes into target mice. Since the up-regulated methylation levels of histone arginine residue lead to the maintenance of pluripotency in embryos and stem cells, it may be suggested that CARM1 overexpressing mESCs elevate the expression of pluripotency-related genes in reconstituted embryos for transgenic mice and may resist the differentiation into trophectoderm (TE). We constructed a fusion protein by connecting CARM1 and 7X-arginine (R7). As a cell-penetrating peptide (CPP), can translocate CARM1 protein into mESCs. CPP-CARM1 protein was detected in the nuclei of the mESCs after a treatment of 24 hours. Accordingly, the expression of pluripotency-related genes was up-regulated in CPP-CARM1-treated mESCs. In addition, CPP-CARM1-treated mESC-derived embryoid bodies (EBs) showed an elevated expression of pluripotency-related genes and delayed spontaneous differentiation. This result suggests that the treatment of recombinant CPP-CARM1 protein elevates the expression of pluripotency-related genes of mESCs by epigenetic modification, and this protein-delivery system could be used to modify embryonic fate in reconstituted embryos with mESCs.

Effect of Water Extract of Eucommiae cortex In RANKL-induced Osteoclast Differentiation (두충의 물 추출물이 파골세포의 분화에 미치는 영향)

  • Jung, Yeon-Tae;Choi, Yun-Hong;Song, Jeong-Hoon;Lee, Chang-Hoon;Lee, Myeung-Su;Jang, Sung-Jo;Cho, Hae-Joong;Kwak, Han-Bok;Oh, Jae-Min
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.3
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    • pp.613-618
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    • 2009
  • Although the effect of Eucommie umoides oliver in osteoporosis has been studied, direct action of Eucommis ulmoides Oliver on osteoclasts remains unknown. Here we examined whether Eucommiae cortex inhibits osteoclast differentiation and bone resorption, a process known to be involved in bone diseases such as osteoporosis. Water extract from Eucommiae cortex (WE-EC) inhibited differentiation of bone marrow macrophages (BMMs) into osteoclasts without causing cytotoxicity. WE-EC suppressed the phosphorylation of p38, ERK, and JNK in BMMs treated with RANKL. WE EC specifically suppressed the mRNA expression of NFATc1 induced by RANKL. However, WE-EC inhibited stability of c-Fos protein induced by RANKL. Furthermore, WE-EC inhibited osteoclast survival induced by RANKL and in turn suppressed bone resorption. Taken together, our results suggest that WE-EC may be better agents for therapeutic use in bone diseases.

Somatic embryogenesis from the axillary meristems of peanut (Arachis hypogaea L.)

  • Singh, Shweta;Hazra, Sulekha
    • Plant Biotechnology Reports
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    • v.3 no.4
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    • pp.333-340
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    • 2009
  • Developmental anomalies in the plumule meristem of peanut (Arachis hypogaea L.) somatic embryos resulted in poor shoot differentiation and reduced plant recovery. Existing meristems with caulogenic potential have never been tested for embryogenesis in peanut. The present experiment was designed to test the mature zygotic embryo axis derived plumule with three meristems for somatic embryogenesis. Embryogenic masses and embryos developed from the caulogenic meristems in the axils. Exposure of 2 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation temporarily which then regained the ability to form the shoot on withdrawal of 2,4-D. Exposure of 4 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation irreversibly. No shoot formation was noted from the tips in any of the cultures which were in secondary medium with $13.6{\mu}M$ 2,4-D. Development of somatic embryos directly from axillary meristems was confirmed histologically. Conversion frequency of these embryos was 11%. Thus, in this report, we describe a method to obtain somatic embryos from the determined organogenic buds of the axillary meristem, by culturing the nodal explant vertically on embryo induction medium. It also displays the possibility of obtaining both embryogenic and organogenic potential in two parts of the same explant simultaneously. The possibility of extending this approach for genetic transformation in in vivo system through direct DNA delivery or Agrobacterium injection in meristems can also be explored. Using Agrobacterium rhizogenes, we have demonstrated the possibility of gene transfer in the axillary meristems of seed-derived plumule explant.

Biocompatibility and Bioactivity of Four Different Root Canal Sealers in Osteoblastic Cell Line MC3T3-El

  • Jun, Nu-Ri;Lee, Sun-Kyung;Lee, Sang-Im
    • Journal of dental hygiene science
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    • v.21 no.4
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    • pp.243-250
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    • 2021
  • Background: Endodontic sealers or their toxic components may become inflamed and lead to delayed wound healing when in direct contact with periapical tissues over an extended period. Moreover, an overfilled sealer can directly interact with adjacent tissues and may cause immediate necrosis or further resorption. Therefore, the treatment outcome conceivably depends on the endodontic sealer's biocompatibility and osteogenic potential. This study aimed to evaluate the cell viability and osteogenic effects of four different sealers in osteoblastic cells. Methods: AH Plus (resin-based sealer), Pulp Canal Sealer EWT (zinc oxide-eugenol sealer), BioRoot RCS (calcium silicate-based sealer), and Well-Root ST (MTA-based calcium silicate sealer) were mixed strictly according to the manufacturer's instructions, and dilutions of sealer extracts (1/2, 1/5 and 1/10) were determined. Cell viability was measured using the water-soluble tetrazolium-8 (WST-8) assay. Differentiation was assessed by alkaline phosphatase (ALP) activity and mineralized nodule formation by Alizarin Red S staining. Results: The cell viability of the extracts derived from the sealers excluding Well-Root ST was concentration dependent, with sealer extracts having the least viability at a 1/2 dilution. At sealer extract dilution of 1/10, the test groups showed the same survival rate as that control group, with the exception of BioRoot RCS. Among all experimental groups, BioRoot RCS showed the highest cell viability after 48 hours. The ALP activity was significantly higher in a concentration-dependent manner. Furthemore, all four materials promoted ALP activity and mineralized nodule formation compared to the control at 1/10 dilutions. Conclusion: This is the first study to highlight the differences in biological activity of these four materials. These results suggest that the composition of root canal sealers appears to alter the form of biocompatibility and osteoblastic differentiation.

Expression of Senescence-Associated Secretory Phenotype in Senescent Gingival Fibroblasts

  • Sangim Lee
    • Journal of dental hygiene science
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    • v.23 no.2
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    • pp.169-175
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    • 2023
  • Background: Although microbial infection is direct cause of periodontal disease, various environmental factors influence the disease severity. Aging is considered a risk factor for oral diseases, with the prevalence of periodontal diseases increasing with age. Moreover, senescence-associated secretory phenotype (SASP) expressed in age-related diseases is a key marker of chronic inflammation and aging phenotypes. Therefore, this study aimed to understand the relevance of senescent cells to periodontal health and disease, investigate the possibility of regulating the expression of aging- and osteolysis-related factors in gingival fibroblasts, and investigate the effect of senescence induction in gingival fibroblasts on osteoclast differentiation in mouse bone marrow-derived macrophages (BMMs). Methods: After stimulation with 400 nM hydrogen peroxidase, human gingival fibroblasts (HGFs) were examined for senescence-associated β-galactosidase. Western blot and enzyme-linked immunosorbent assays were performed to assess the expression of SASP. Osteoclast formation was assessed in BMMs using a conditioned medium (CM) from hydrogen peroxide-stimulated HGFs. Osteoclastic differentiation was investigated using tartrate-resistant acid phosphatase (TRAP) staining and activity. Data analysis was performed using SPSS version 25.0. Results: The expression of senescence-related molecules, including p53, p16, and p21, and the expression of osteolytic factors, including IL-6, IL-8, and IL-17, were found to be significantly higher in the hydrogen peroxide-stimulated HGF than in the control group. Regarding the indirect effects of senescent gingival cells, the number of osteoclasts and TRAP activity increased according to the differentiation of BMM cultured in CM. Conclusion: Our results on the of between osteolytic factors and cellular senescence in gingival fibroblast cells helped to reveal evidence of pathological aging mechanisms. Furthermore, our results suggest that the development of novel therapies that target specific SASP factors could be an effective treatment strategy for periodontal disease.

Tracking of Stem Cells from Human Exfoliated Deciduous Teeth Labeled with Molday ION Rhodamine-B during Periodontal Bone Regeneration in Rats

  • Nan Zhang;Li Xu;Hao Song;Chunqing Bu;Jie Kang;Chuanchen Zhang;Xiaofei Yang;Fabin Han
    • International Journal of Stem Cells
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    • v.16 no.1
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    • pp.93-107
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    • 2023
  • Background and Objectives: Chronic periodontitis can lead to alveolar bone resorption and eventually tooth loss. Stem cells from exfoliated deciduous teeth (SHED) are appropriate bone regeneration seed cells. To track the survival, migration, and differentiation of the transplanted SHED, we used super paramagnetic iron oxide particles (SPIO) Molday ION Rhodamine-B (MIRB) to label and monitor the transplanted cells while repairing periodontal bone defects. Methods and Results: We determined an appropriate dose of MIRB for labeling SHED by examining the growth and osteogenic differentiation of labeled SHED. Finally, SHED was labeled with 25 ㎍ Fe/ml MIRB before being transplanted into rats. Magnetic resonance imaging was used to track SHED survival and migration in vivo due to a low-intensity signal artifact caused by MIRB. HE and immunohistochemical analyses revealed that both MIRB-labeled and unlabeled SHED could promote periodontal bone regeneration. The colocalization of hNUC and MIRB demonstrated that SHED transplanted into rats could survive in vivo. Furthermore, some MIRB-positive cells expressed the osteoblast and osteocyte markers OCN and DMP1, respectively. Enzyme-linked immunosorbent assay revealed that SHED could secrete protein factors, such as IGF-1, OCN, ALP, IL-4, VEGF, and bFGF, which promote bone regeneration. Immunofluorescence staining revealed that the transplanted SHED was surrounded by a large number of host-derived Runx2- and Col II-positive cells that played important roles in the bone healing process. Conclusions: SHED could promote periodontal bone regeneration in rats, and the survival of SHED could be tracked in vivo by labeling them with MIRB. SHED are likely to promote bone healing through both direct differentiation and paracrine mechanisms.

Back Analysis Method for Material Properties of Multi-layers Ground Considering Multiple Unknown Variables (다중 미지변수를 고려한 다층지반 역해석)

  • Kim, Se-Jin;Kim, Moon-Kyum;Won, Jong-Hwa;Kim, Jung-Soo
    • Journal of the Korean Geotechnical Society
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    • v.25 no.9
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    • pp.93-100
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    • 2009
  • A core procedure of the direct search method used in this study is optimizing a difference between objective function and real displacement and correcting unknown variables. Because the research procedure comes from back-analyzing of the unknown variable of each layer, back-analyzing results need an additional optimization to minimize interferential effects of unknown variables. Therefore, the direct search method Is used to obtain optimized solutions without a partial differentiation of an objective function. The object of this research is developing the back analysis technique for multi-unknown variables by modeling the soil including underground structure Into upper and lower layer. In order to minimize interferent errors, repeated back analysis is performed and applicability on the real tunnel is examined. Consequently, the multi-layer analysis model is more precise in describing the real behavior of underground structure. It shows the validity of back analysis far multi-layer model which is the understructure placed on multi-layer boundaries.

An Overview of Optimization of Structures Subjected to Transient Loads (동하중을 받는 구조물의 최적화에 관한 연구동향)

  • Park Gyung-Jin;Kang Byung-Soo
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.29 no.3 s.234
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    • pp.369-386
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    • 2005
  • Various aspects of structural optimization techniques under transient loads are extensively reviewed. The main themes of the paper are treatment of time dependent constraints, calculation of design sensitivity, and approximation. Each subject is reviewed with the corresponding papers that have been published since 1970s. The treatment of time dependent constraints in both the direct method and the transformation method is discussed. Two ways of calculating design sensitivity of a structure under transient loads are discussed - direct differentiation method and adjoint variable method. The approximation concept mainly focuses on re- sponse surface method in crashworthiness and local approximation with the intermediate variable Especially, as an approximated optimization technique, Equivalent Static Load method which takes advantage of the well-established static response optimization technique is introduced. And as an application area of dynamic response optimization technique, the structural optimization in flexible multibody dynamic systems is re- viewed in the viewpoint of the above three themes