• Title/Summary/Keyword: molecular processes

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The Integrins Involved in Soybean Agglutinin-Induced Cell Cycle Alterations in IPEC-J2

  • Pan, Li;Zhao, Yuan;Yuan, Zhijie;Farouk, Mohammed Hamdy;Zhang, Shiyao;Bao, Nan;Qin, Guixin
    • Molecules and Cells
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    • v.40 no.2
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    • pp.109-116
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    • 2017
  • Soybean agglutinin (SBA) is an anti-nutritional factor of soybean, affecting cell proliferation and inducing cytotoxicity. Integrins are transmembrane receptors, mediating a variety of cell biological processes. This research aims to study the effects of SBA on cell proliferation and cell cycle progression of the intestinal epithelial cell line from piglets (IPEC-J2), to identify the integrin subunits especially expressed in IPEC-J2s, and to analyze the functions of these integrins on IPEC-J2 cell cycle progression and SBA-induced IPEC-J2 cell cycle alteration. The results showed that SBA lowered cell proliferation rate as the cell cycle progression from G0/G1 to S phase (P < 0.05) was inhibited. Moreover, SBA lowered mRNA expression of cell cycle-related gene CDK4, Cyclin E and Cyclin D1 (P < 0.05). We successfully identified integrins ${\alpha}2$, ${\alpha}3$, ${\alpha}6$, ${\beta}1$, and ${\beta}4$ in IPEC-J2s. These five subunits were crucial to maintain normal cell proliferation and cell cycle progression in IPEC-J2s. Restrain of either these five subunits by their inhibitors, lowered cell proliferation rate, and arrested the cells at G0/G1 phase of cell cycle (P < 0.05). Further analysis indicated that integrin ${\alpha}2$, ${\alpha}6$, and ${\beta}1$ were involved in the blocking of G0/G1 phase induced by SBA. In conclusion, these results suggested that SBA lowered the IPEC-J2 cell proliferation rate through the perturbation of cell cycle progression. Furthermore, integrins were important for IPEC-J2 cell cycle progression, and they were involved in the process of SBA-induced cell cycle progression alteration, which provide a basis for further revealing SBA anti-proliferation and anti-nutritional mechanism.

Modification of Water-borne Polyurethane Using Benzophenone Crosslinker (Benzophenone 가교제를 이용한 수분산 폴리우레탄 개질)

  • Kim, HyeokJin;Kim, Jin Chul;Chang, SangMok;Seo, BongKuk
    • Applied Chemistry for Engineering
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    • v.27 no.2
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    • pp.221-226
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    • 2016
  • Production of eco-friendly and biologically harmless materials is strongly required in all industries. In particular, reducing volatile organic compounds in coating processes is extremely important to secure worker's safety. During recent two decades, extensive research works on water-borne polyurethane dispersion (PUD) have been continuously developed as an alternative to solvent-borne polyurethane. However, PUD was shown inferior mechanical properties to the organic solvent-borne polyurethane due to a limit to the molecular weight increase, which resulted in the limit of applications. To overcome this drawback, several approaches have been examined such as polymer blends and thermal/radiation induced crosslinking. Among these methods, the radiation curing system was suitable for industrialization because of the high crosslinking density and fast curing speed. In this study, we overcame the drawback for PUD via introducing benzophenone radiation curable units to PUD. We synthesized PUD films which possessed good dispersion in water for 30 days, increased Tg and Td more than $5^{\circ}C$ after UV curing film as well as improved young's modulus more than double.

Expression and Characterization of Fibrinolytic Enzyme Activity During Earthworm Tail Regeneration (지렁이 꼬리 재생시 발현되는 피브리노겐 분해효소의 활성과 특성에 관한 연구)

  • Tak, Eun-Sik;Cho, Sung-Jin;Kim, Jae-Young;Lee, Kyu-Seok;Park, Soon-Cheol
    • The Korean Journal of Soil Zoology
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    • v.4 no.2
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    • pp.101-106
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    • 1999
  • Fibrinolytic enzyme is thought to be involved in extracellular matrix remodeling during regeneration. We investigated the expression and characterization fibrinolytic enzyme activity during earthworm tail regeneration. Electrophoretic analysis of fibrinolytic enzymes induced during regeneration revealed that at least seven types of fibrinolytic enzymes were expressed, which had molecular weight of 12, 19, 23, 27, 32, 45 and 58 kDa, respectively. These fibrinolytic enzyme activities were dramatically increased within 1 day after amputation. These activities were maintained by 7 days postamputation, followed by decrease to control level from 14 days after amputation. Alltypes of fibrinolytic enzyme activities were inhibited by treatment of PMSF and aprotinin, and were insensitive to EDTA and exogenous Ca$^{2+}$. These results indicate that the fibrinolytic enzymes are serineproteinase. Other characteristics including specificities for extracellular matrix proteins are under investigation. Based on these results, we are trying to find out the relationship among expression of proteinases, extracellular matrix remodeling, and dedifferentiation, which are believed to be essential processes during regeneration.

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Ubiquitin D Promotes Progression of Oral Squamous Cell Carcinoma via NF-Kappa B Signaling

  • Song, An;Wang, Yi;Jiang, Feng;Yan, Enshi;Zhou, Junbo;Ye, Jinhai;Zhang, Hongchuang;Ding, Xu;Li, Gang;Wu, Yunong;Zheng, Yang;Song, Xiaomeng
    • Molecules and Cells
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    • v.44 no.7
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    • pp.468-480
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    • 2021
  • Ubiquitin D (UBD) is highly upregulated in many cancers, and plays a pivotal role in the pathophysiological processes of cancers. However, its roles and underlying mechanisms in oral squamous cell carcinoma (OSCC) are still unclear. In the present study, we investigated the role of UBD in patients with OSCC. Quantitative real-time polymerase chain reaction and Western blot were used to measure the expression of UBD in OSCC tissues. Immunohistochemistry assay was used to detect the differential expressions of UBD in 244 OSCC patients and 32 cases of normal oral mucosae. In addition, CCK-8, colony formation, wound healing and Transwell assays were performed to evaluate the effect of UBD on the cell proliferation, migration, and invasion in OSCC. Furthermore, a xenograft tumor model was established to verify the role of UBD on tumor formation in vivo. We found that UBD was upregulated in human OSCC tissues and cell lines and was associated with clinical and pathological features of patients. Moreover, the overexpression of UBD promoted the proliferation, migration and invasion of OSCC cells; however, the knockdown of UBD exerted the opposite effects. In this study, our results also suggested that UBD promoted OSCC progression through NF-κB signaling. Our findings indicated that UBD played a critical role in OSCC and may serve as a prognostic biomarker and potential therapeutic target for OSCC treatment.

Exocyclic GpC DNA methyltransferase from Celeribacter marinus IMCC12053 (Celeribacter marinus IMCC12053의 외향고리 GpC DNA 메틸트랜스퍼라아제)

  • Kim, Junghee;Oh, Hyun-Myung
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.103-111
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    • 2019
  • DNA methylation is involved in diverse processes in bacteria, including maintenance of genome integrity and regulation of gene expression. CcrM, the DNA methyltransferase conserved in Alphaproteobacterial species, carries out $N^6$-adenine or $N^4$-cytosine methyltransferase activities using S-adenosyl methionine as a co-substrate. Celeribacter marinus IMCC12053 from the Alphaproteobacterial group was isolated from a marine environment. Single molecule real-time sequencing method (SMRT) was used to detect the methylation patterns of C. marinus IMCC12053. Gibbs motif sampler program was used to observe the conversion of adenosine of 5'-GANTC-3' to $N^6$-methyladenosine and conversion of $N^4$-cytosine of 5'-GpC-3' to $N^4$-methylcytosine. Exocyclic DNA methyltransferase from the genome of strain IMCC12053 was chosen using phylogenetic analysis and $N^4$-cytosine methyltransferase was cloned. IPTG inducer was used to confirm the methylation activity of DNA methylase, and cloned into a pQE30 vector using dam-/dcm- E. coli as the expression host. The genomic DNA and the plasmid carrying methylase-encoding sequences were extracted and cleaved with restriction enzymes that were sensitive to methylation, to confirm the methylation activity. These methylases protected the restriction enzyme site once IPTG-induced methylases methylated the chromosome and plasmid, harboring the DNA methylase. In this study, cloned exocyclic DNA methylases were investigated for potential use as a novel type of GpC methylase for molecular biology and epigenetics.

Evaluation of the Effectiveness of Surveillance on Improving the Detection of Healthcare Associated Infections (의료관련감염에서 감시 개선을 위한 평가)

  • Park, Chang-Eun
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.1
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    • pp.15-25
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    • 2019
  • The development of reliable and objective definitions as well as automated processes for the detection of health care-associated infections (HAIs) is crucial; however, transformation to an automated surveillance system remains a challenge. Early outbreak identification usually requires clinicians who can recognize abnormal events as well as ongoing disease surveillance to determine the baseline rate of cases. The system screens the laboratory information system (LIS) data daily to detect candidates for health care-associated bloodstream infection (HABSI) according to well-defined detection rules. The system detects and reserves professional autonomy by requiring further confirmation. In addition, web-based HABSI surveillance and classification systems use discrete data elements obtained from the LIS, and the LIS-provided data correlates strongly with the conventional infection-control personnel surveillance system. The system was timely, acceptable, useful, and sensitive according to the prevention guidelines. The surveillance system is useful because it can help health care professionals better understand when and where the transmission of a wide range of potential pathogens may be occurring in a hospital. A national plan is needed to strengthen the main structures in HAI prevention, Healthcare Associated Prevention and Control Committee (HAIPCC), sterilization service (SS), microbiology laboratories, and hand hygiene resources, considering their impact on HAI prevention.

A chemical conjugate between HER2-targeting antibody fragment and Pseudomonas exotoxin A fragment demonstrates cytotoxic effects on HER2-expressing breast cancer cells

  • Lee, Sunju;Park, Sangsu;Nguyen, Minh Tan;Lee, Eunyoung;Kim, Julee;Baek, Sangki;Kim, Chong Jai;Jang, Yeon Jin;Choe, Han
    • BMB Reports
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    • v.52 no.8
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    • pp.496-501
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    • 2019
  • Conventionally, immunotoxins have been produced as a single polypeptide from fused genes of an antibody fragment and a toxin. In this study, we adopted a unique approach of chemical conjugation of a toxin protein and an antibody fragment. The two genes were separately expressed in Escherichia coli and purified to high levels of purity. The two purified proteins were conjugated using a chemical linker. The advantage of this approach is its ability to overcome the problem of low recombinant immunotoxin production observed in some immunotoxins. Another advantage is that various combinations of immunotoxins can be prepared with fewer efforts, because the chemical conjugation of components is relatively simpler than the processes involved in cloning, expression, and purification of multiple immunotoxins. As a proof of concept, the scFv of trastuzumab and the PE24 fragment of Pseudomonas exotoxin A were separately produced using E. coli and then chemically crosslinked. The new immunotoxin was tested on four breast cancer cell lines variably expressing HER2. The chemically crosslinked immunotoxin exhibited cytotoxicity in proportion to the expression level of HER2. In conclusion, the present study revealed an alternative method of generating an immunotoxin that could effectively reduce the viability of HER2-expressing breast cancer cells. These results suggest the effectiveness of this method of immunotoxin crosslinking as a suitable alternative for producing immunotoxins.

Inhibition of MicroRNA-15a/16 Expression Alleviates Neuropathic Pain Development through Upregulation of G Protein-Coupled Receptor Kinase 2

  • Li, Tao;Wan, Yingchun;Sun, Lijuan;Tao, Shoujun;Chen, Peng;Liu, Caihua;Wang, Ke;Zhou, Changyu;Zhao, Guoqing
    • Biomolecules & Therapeutics
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    • v.27 no.4
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    • pp.414-422
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    • 2019
  • There is accumulating evidence that microRNAs are emerging as pivotal regulators in the development and progression of neuropathic pain. MicroRNA-15a/16 (miR-15a/16) have been reported to play an important role in various diseases and inflammation response processes. However, whether miR-15a/16 participates in the regulation of neuroinflammation and neuropathic pain development remains unknown. In this study, we established a mouse model of neuropathic pain by chronic constriction injury (CCI) of the sciatic nerves. Our results showed that both miR-15a and miR-16 expression was significantly upregulated in the spinal cord of CCI rats. Downregulation of the expression of miR-15a and miR-16 by intrathecal injection of a specific inhibitor significantly attenuated the mechanical allodynia and thermal hyperalgesia of CCI rats. Furthermore, inhibition of miR-15a and miR-16 downregulated the expression of interleukin-$1{\beta}$ and tumor-necrosis factor-${\alpha}$ in the spinal cord of CCI rats. Bioinformatic analysis predicted that G protein-coupled receptor kinase 2 (GRK2), an important regulator in neuropathic pain and inflammation, was a potential target gene of miR-15a and miR-16. Inhibition of miR-15a and miR-16 markedly increased the expression of GRK2 while downregulating the activation of p38 mitogen-activated protein kinase and $NF-{\kappa}B$ in CCI rats. Notably, the silencing of GRK2 significantly reversed the inhibitory effects of miR-15a/16 inhibition in neuropathic pain. In conclusion, our results suggest that inhibition of miR-15a/16 expression alleviates neuropathic pain development by targeting GRK2. These findings provide novel insights into the molecular pathogenesis of neuropathic pain and suggest potential therapeutic targets for preventing neuropathic pain development.

Current Perspectives on the Effects of Plant Growth-promoting Rhizobacteria (식물생장촉진 근권미생물의 영향에 대한 연구 현황 및 전망)

  • Le, Thien Tu Huynh;Jun, Sang Eun;Kim, Gyung-Tae
    • Journal of Life Science
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    • v.29 no.11
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    • pp.1281-1293
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    • 2019
  • The rhizosphere is the active zone where plant roots communicate with the soil microbiome, each responding to the other's signals. The soil microbiome within the rhizosphere that is beneficial to plant growth and productivity is known as plant growth-promoting rhizobacteria (PGPR). PGPR take part in many pivotal plant processes, including plant growth, development, immunity, and productivity, by influencing acquisition and utilization of nutrient molecules, regulation of phytohormone biosynthesis, signaling, and response, and resistance to biotic- and abiotic-stresses. PGPR also produce secondary compounds and volatile organic compounds (VOCs) that elicit plant growth. Moreover, plant roots exude attractants that cause PGPR to aggregate in the rhizosphere zone for colonization, improving soil properties and protecting plants against pathogenic factors. The interactions between PGPR and plant roots in rhizosphere are essential and interdependent. Many studies have reported that PGPR function in multiple ways under the same or diverse conditions, directly and indirectly. This review focuses on the roles and strategies of PGPR in enhancing nutrient acquisition by nutrient fixation/solubilization/mineralization, inducing plant growth regulators/phytohormones, and promoting growth and development of root and shoot by affecting cell division, elongation, and differentiation. We also summarize the current knowledge of the effects of PGPR and the soil microbiota on plants.

Outbreak of Fire Blight of Apple and Pear and Its Characteristics in Korea in 2019 (2019년 국내 사과와 배 화상병 대발생과 그 특징)

  • Ham, Hyeonheui;Lee, Kyong Jae;Hong, Seong Jun;Kong, Hyun Gi;Lee, Mi-Hyun;Kim, Hyun-Ran;Lee, Yong Hwan
    • Research in Plant Disease
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    • v.26 no.4
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    • pp.239-249
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    • 2020
  • To find out the cause of the fire blight outbreak in apples and pears of Korea in 2019, we investigated disease appearing situation of thirty fire blight infected orchards, and interviewed farmers to determine the cultivation characteristics. Fire blight occurred mostly in orchards that had infected more than 2 years before. The cause of this were as follows: farmers did not know the symptoms of the disease properly. It is presumed that it has spread from the first occurrence to the surrounding orchards by flower-visiting insects or farmers and to a short distance or a long distance by the same cultivator or co-farmer. These series of processes repeated in the newly spreading area, and then disease reports increased as farmers became aware of fire blight. To minimize the spread of fire blight in Korea, it suggested that thorough education of farmers for early diagnosis and quantitative detection technology that can diagnose even in no symptom showing plants. And chemical or biological spraying systems suitable for domestic cultivation methods, which are producing large fruits, and molecular epidemiological studies of pathogens.