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An ARIA-Interacting AP2 Domain Protein Is a Novel Component of ABA Signaling

  • Lee, Sun-ji;Cho, Dong-im;Kang, Jung-youn;Kim, Soo Young
    • Molecules and Cells
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    • v.27 no.4
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    • pp.409-416
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    • 2009
  • ADAP is an AP2-domain protein that interacts with ARIA, which, in turn, interacts with ABF2, a bZIP class transcription factor. ABF2 regulates various aspects of the abscisic acid (ABA) response by controlling the expression of a subset of ABA-responsive genes. Our expression analyses indicate that ADAP is expressed in roots, emerging young leaves, and flowers. We found that adap knockout mutant lines germinate more efficiently than wild-type plants and that the mutant seedlings grow faster. This suggests that ADAP is involved in the regulation of germination and seedling growth. Both germination and post-germination growth of the knockout mutants were partially insensitive to ABA, which indicates that ADAP is required for a full ABA response. The survival rates for mutants from which water was withheld were low compared with those for wild-type plants. The result shows that ADAP is necessary for the response to stress induced by water deprivation. Together, our data indicate that ADAP is a positive regulator of the ABA response and is also involved in regulating seedling growth. The role of ADAP is similar to that of ARIA, which is also a positive regulator of the ABA response. It appears that ADAP acts through the same ABA response pathway as ARIA.

The p53-p21Cip1/WAF1 Pathway Is Necessary for Cellular Senescence Induced by the Inhibition of Protein Kinase CKII in Human Colon Cancer Cells

  • Kang, Ji-Young;Kim, Jin Joo;Jang, Seok Young;Bae, Young-Seuk
    • Molecules and Cells
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    • v.28 no.5
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    • pp.489-494
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    • 2009
  • We have previously shown that the down-regulation of protein kinase CKII activity is tightly associated with cellular senescence of human fibroblast IMR-90 cells. Here, we examined the roles of p53 and $p21^{Cip1/WAF1}$ in senescence development induced by CKII inhibition using wild-type, isogenic p53-/- and isogenic p21-/- HCT116 human colon cancer cell lines. A senescent marker appeared after staining for senescence-associated ${\beta}$-galactosidase activity in wild-type HCT116 cells treated with CKII inhibitor or $CKII{\alpha}$ siRNA, but this response was almost abolished in p53- or $p21^{Cip1/WAF1}$-null cells. Increased cellular levels of p53 and $p21^{Cip1/WAF1}$ protein occurred with the inhibition of CKII. CKII inhibition upregulated p53 and $p21^{Cip1/WAF1}$ expression at post-transcriptional level and transcription level, respectively. RB phosphorylation significantly decreased in cells treated with CKII inhibitor. Taken together, this study shows that the activation of the $p53-p21^{Cip1/WAF1}$ pathway acts as a major mediator of cellular senescence induced by CKII inhibition.

Generation of a recombinant rabies virus expressing green fluorescent protein for a virus neutralization antibody assay

  • Yang, Dong-Kun;Kim, Ha-Hyun;Park, Yu-Ri;Yoo, Jae Young;Park, Yeseul;Park, Jungwon;Hyun, Bang-Hun
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.56.1-56.10
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    • 2021
  • Background: Fluorescent antibody virus neutralization (FAVN) test is a standard assay for quantifying rabies virus-neutralizing antibody (VNA) in serum. However, a safer rabies virus (RABV) should be used in the FAVN assay. There is a need for a new method that is economical and time-saving by eliminating the immunostaining step. Objectives: We aimed to improve the traditional FAVN method by rescuing and characterizing a new recombinant RABV expressing green fluorescent protein (GFP). Methods: A new recombinant RABV expressing GFP designated as ERAGS-GFP was rescued using a reverse genetic system. Immuno-fluorescence assay, peroxidase-linked assay, electron microscopy and reverse transcription polymerase chain reaction were performed to confirm the recombinant ERAGS-GFP virus as a RABV expressing the GFP gene. The safety of ERAGS-GFP was evaluated in 4-week-old mice. The rabies VNA titers were measured and compared with conventional FAVN and FAVN-GFP tests using VERO cells. Results: The virus propagated in VERO cells was confirmed as RABV expressing GFP. The ERAGS-GFP showed the highest titer (108.0 TCID50/mL) in VERO cells at 5 days post-inoculation, and GFP expression persisted until passage 30. The body weight of 4-week-old mice inoculated intracranially with ERAGS-GFP continued to increase and the survival rate was 100%. In 62 dog sera, the FAVN-GFP result was significantly correlated with that of conventional FAVN (r = 0.95). Conclusions: We constructed ERAGS-GFP, which could replace the challenge virus standard-11 strain used in FAVN test.

Effects of the cathepsin K inhibitor with mineral trioxide aggregate cements on osteoclastic activity

  • Kim, Hee-Sun;Kim, Soojung;Ko, Hyunjung;Song, Minju;Kim, Miri
    • Restorative Dentistry and Endodontics
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    • v.44 no.2
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    • pp.17.1-17.10
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    • 2019
  • Objectives: Root resorption is an unexpected complication after replantation procedures. Combining anti-osteoclastic medicaments with retrograde root filling materials may avert this resorptive activity. The purpose of this study was to assess effects of a cathepsin K inhibitor with calcium silicate-based cements on osteoclastic activity. Methods: MC3T3-E1 cells were cultured for biocompatibility analyses. RAW 264.7 cells were cultured in the presence of the receptor activator of nuclear factor-kappa B and lipopolysaccharide, followed by treatment with Biodentine (BIOD) or ProRoot MTA with or without medicaments (Odanacatib [ODN], a cathepsin inhibitor and alendronate, a bisphosphonate). After drug treatment, the cell counting kit-8 assay and Alizarin red staining were performed to evaluate biocompatibility in MC3T3-E1 cells. Reverse-transcription polymerase chain reaction, tartrate-resistant acid phosphatase (TRAP) staining and enzyme-linked immunosorbent assays were performed in RAW 264.7 cells to determine the expression levels of inflammatory cytokines, interleukin $(IL)-1{\beta}$, IL-6, tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and prostaglandin E2 (PGE2). Data were analyzed by one-way analysis of variance and Tukey's post hoc test (p < 0.05). Results: Biocompatibility results showed that there were no significant differences among any of the groups. RAW 264.7 cells treated with BIOD and ODN showed the lowest levels of $TNF-{\alpha}$ and PGE2. Treatments with BIOD + ODN were more potent suppressors of inflammatory cytokine expression (p < 0.05). Conclusion: The cathepsin K inhibitor with calcium silicate-based cement inhibits osteoclastic activity. This may have clinical application in preventing inflammatory root resorption in replanted teeth.

Adverse Birth Outcomes Among Pregnant Women With and Without COVID-19: A Comparative Study From Bangladesh

  • Masud, Sumaya Binte;Zebeen, Faiza;Alam, Dil Ware;Hossian, Mosharap;Zaman, Sanjana;Begum, Rowshan Ara;Nabi, Mohammad Hayatun;Hawlader, Mohammad Delwer Hossain
    • Journal of Preventive Medicine and Public Health
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    • v.54 no.6
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    • pp.422-430
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    • 2021
  • Objectives: Pregnant women are especially vulnerable to respiratory infections such as coronavirus disease 2019 (COVID-19), but insufficient research has investigated pregnancy and its outcomes in women with COVID-19. This cross-sectional study compared birth outcomes related to COVID-19 between Bangladeshi pregnant women with and without COVID-19. Methods: The study was conducted at 3 tertiary referral hospitals in Dhaka, Bangladesh, from March to August 2020. Pregnant women admitted for delivery at these hospitals with laboratory results (reverse-transcription polymerase chain reaction) were analyzed. Using convenience sampling, we included 70 COVID-19-positive and 140 COVID-19-negative pregnant women. Trained and experienced midwives conducted the interviews. Data were analyzed using the t-test, the chi-square test, and univariate and multivariable linear and logistic regression. Results: Pregnant women with COVID-19 were more likely to give birth to a preterm baby (adjusted odds ratio [aOR], 2.15; 95% confidence interval [CI], 1.06 to 4.37) and undergo a cesarean section (aOR, 3.27; 95% CI, 1.51 to 7.07). There were no significant differences in birth weight, premature rupture of membranes, and the Apgar score at 1 minute or 5 minutes post-delivery between women with and without COVID-19. All the newborn babies who were born to COVID-19-positive women were COVID-19-negative. Conclusions: Our study suggests that pregnant women with COVID-19 were more likely to give birth to a preterm baby and undergo a cesarean section. For this reason, physicians should be particularly cautious to minimize adverse birth outcomes among pregnant women with COVID-19 and their newborn babies.

Analyzing the changes of elementary pre-service teacher's noticing occurred in the process of participating in the actual class and learning community (실제 수업 및 학습공동체 참여 과정에서 나타난 초등 예비교사의 주목하기 변화 분석)

  • Jung, Hye-Yun;Seo, Yumin;Han, Jooho;Seo, Minju
    • Journal of the Korean School Mathematics Society
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    • v.25 no.3
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    • pp.279-306
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    • 2022
  • The purpose of this case study is to get an implication on elementary pre-service teacher education programs by exploring how a pre-service teacher's noticing changes within a learning community. The pre-service teacher participated in a learning community with researchers. Data includes recordings and transcription of actual class and pre- and post discussion in the learning community, the pre-service teacher's reflection essays, field notes, and students' worksheets. Results are as follows. First, the pre-service teacher's attending moved from the result of tasks to students' mathematical thinking. Second, the pre-service teacher's interpretation changed from a lack of diversity and specificity of evidence to diversity and specificity. Third, the pre-service teacher's decision-makings changed from unproductive deciding to productive deciding.

Differences in liver microRNA profiling in pigs with low and high feed efficiency

  • Miao, Yuanxin;Fu, Chuanke;Liao, Mingxing;Fang, Fang
    • Journal of Animal Science and Technology
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    • v.64 no.2
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    • pp.312-329
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    • 2022
  • Feed cost is the main factor affecting the economic benefits of pig industry. Improving the feed efficiency (FE) can reduce the feed cost and improve the economic benefits of pig breeding enterprises. Liver is a complex metabolic organ which affects the distribution of nutrients and regulates the efficiency of energy conversion from nutrients to muscle or fat, thereby affecting feed efficiency. MicroRNAs (miRNAs) are small non-coding RNAs that can regulate feed efficiency through the modulation of gene expression at the post-transcriptional level. In this study, we analyzed miRNA profiling of liver tissues in High-FE and Low-FE pigs for the purpose of identifying key miRNAs related to feed efficiency. A total 212~221 annotated porcine miRNAs and 136~281 novel miRNAs were identified in the pig liver. Among them, 188 annotated miRNAs were co-expressed in High-FE and Low-FE pigs. The 14 miRNAs were significantly differentially expressed (DE) in the livers of high-FE pigs and low-FE pigs, of which 5 were downregulated and 9 were upregulated. Kyoto Encyclopedia of Genes and Genomes analysis of liver DE miRNAs in high-FE pigs and low-FE pigs indicated that the target genes of DE miRNAs were significantly enriched in insulin signaling pathway, Gonadotropin-releasing hormone signaling pathway, and mammalian target of rapamycin signaling pathway. To verify the reliability of sequencing results, 5 DE miRNAs were randomly selected for quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The qRT-PCR results of miRNAs were confirmed to be consistent with sequencing data. DE miRNA data indicated that liver-specific miRNAs synergistically acted with mRNAs to improve feed efficiency. The liver miRNAs expression analysis revealed the metabolic pathways by which the liver miRNAs regulate pig feed efficiency.

Effect of Astragali Radix on SPS-induced PTSD in Rats (황기(黃芪)가 흰쥐에서 SPS로 유도된 PTSD에 미치는 효과)

  • Min, Ye-Eun;Lee, Tae-Hee
    • Herbal Formula Science
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    • v.30 no.3
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    • pp.109-121
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    • 2022
  • Objective : This study is conducted to investigate the effect of Astragali Radix on Post Traumatic Stress Disorder(PTSD) induced by the Single Proposed Stress(SPS). Methods : The experiment was conducted with five groups; SAL groups with only saline treatment, SPS group, SPS + ARX25 group, SPS + ARX50 group, and SPS + ARX100 group. After applying SPS, saline and ARX were administered for 14 days to identify the change of body weight, sucrose intake amount, and behavioral changes through Open Field Test(OFT) and Forced Swimming Test(FST). After the behavioral experiment, plasma corticosterone levels, serotonin, norepinephrine and dopamine concentrations were measured by enzyme-linked immunoassay in medical prefrontal cortex, hippocampus, and amygdala. Brain-derived neurotrophic factor(BDNF) in the hippocampus was measured using Reverse Transcription Polymerase Chain Reaction. Results : Weight change has significantly decreased in the SPS group compared to the SAL group(p<0.05). On day 14, the sucrose intake of rats has significantly increased in the SPS + ARX100 group compared to the SPS group(p<0.05). In OFT, the number of staying in the central space has significantly increased in the SPS + ARX100 group(p<0.01). In FST, immobility has significantly decreased in SPS + ARX50 group and SPS + ARX100 group(p<0.05). The concentration of serotonine, dopamine and BDNF expression has increased significantly in SPS + ARX100 group compared to SPS group(p<0.05) Conclusions : In the SPS-induced PTSD experiment, ARX increased sucrose intake and the numbers of crossing in the central zone space in OFT, decreased immobility time in FST, and increased concentration of serotonin, dopamine, and BDNF. It can be postulated that the ARX could be effective for the treatment of PTSD.

Effects of CTHRC1 on odontogenic differentiation and angiogenesis in human dental pulp stem cells

  • Jong-soon Kim;Bin-Na Lee;Hoon-Sang Chang;In-Nam Hwang;Won-Mann Oh;Yun-Chan Hwang
    • Restorative Dentistry and Endodontics
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    • v.48 no.2
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    • pp.18.1-18.10
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    • 2023
  • Objectives: This study aimed to determine whether collagen triple helix repeat containing-1 (CTHRC1), which is involved in vascular remodeling and bone formation, can stimulate odontogenic differentiation and angiogenesis when administered to human dental pulp stem cells (hDPSCs). Materials and Methods: The viability of hDPSCs upon exposure to CTHRC1 was assessed with the WST-1 assay. CTHRC1 doses of 5, 10, and 20 ㎍/mL were administered to hDPSCs. Reverse-transcription polymerase reaction was used to detect dentin sialophosphoprotein, dentin matrix protein 1, vascular endothelial growth factor, and fibroblast growth factor 2. The formation of mineralization nodules was evaluated using Alizarin red. A scratch wound assay was conducted to evaluate the effect of CTHRC1 on cell migration. Data were analyzed using 1-way analysis of variance followed by the Tukey post hoc test. The threshold for statistical significance was set at p < 0.05. Results: CTHRC1 doses of 5, 10, and 20 ㎍/mL had no significant effect on the viability of hDPSCs. Mineralized nodules were formed and odontogenic markers were upregulated, indicating that CTHRC1 promoted odontogenic differentiation. Scratch wound assays demonstrated that CTHRC1 significantly enhanced the migration of hDPSCs. Conclusions: CTHRC1 promoted odontogenic differentiation and mineralization in hDPSCs.

Expression of Toll-like receptors 3, 7, 9 and cytokines in feline infectious peritonitis virus-infected CRFK cells and feline peripheral monocytes

  • Khair, Megat Hamzah Megat Mazhar;Selvarajah, Gayathri Thevi;Omar, Abdul Rahman;Mustaffa-Kamal, Farina
    • Journal of Veterinary Science
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    • v.23 no.2
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    • pp.27.1-27.16
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    • 2022
  • Background: The role of Toll-like receptors (TLRs) in a feline infectious peritonitis virus (FIPV) infection is not completely understood. Objectives: This study examined the expression of TLR3, TLR7, TLR9, tumor necrosis factor-alpha (TNF-α), interferon (IFN)-β, and interleukin (IL)-10 upon an FIPV infection in Crandell-Reese feline kidney (CRFK) cells and feline monocytes. Methods: CRFK cells and monocytes from feline coronavirus (FCoV)-seronegative cats and FCoV-seropositive cats were infected with type II FIPV-79-1146. At four, 12, and 24 hours post-infection (hpi), the expression of TLR3, TLR7, TLR9, TNF-α, IFN-β, and IL-10, and the viral load were measured using reverse transcription quantitative polymerase chain reaction. Viral protein production was confirmed using immunofluorescence. Results: FIPV-infected CRFK showed the upregulation of TLR9, TNF-α, and IFN-β expression between 4 and 24 hpi. Uninfected monocytes from FCoV-seropositive cats showed lower TLR3 and TLR9 expression but higher TLR7 expression compared to uninfected monocytes from FCoV-seronegative cats. FIPV-infected monocytes from FCoV-seropositive cats downregulated TLR7 and TNF-α expression between 4 and 24 hpi, and 4 and 12 hpi, respectively. IFN-β was upregulated early in FIPV-infected monocytes from FCoV-seropositive cats, with a significant difference observed at 12 hpi compared to FCoV-seronegative cats. The viral load in the CRFK and FIPV-infected monocytes in both cohorts of cats was similar over time.ConclusionTLR7 may be the key TLR involved in evading the innate response against inhibiting TNF-α production. Distinct TLR expression profiles between FCoV-seronegative and FCoV-seropositive cats were observed. The associated TLR that plays a role in the induction of IFN-β needs to be explored further.