• Title/Summary/Keyword: Glycine receptor

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The Progression of SARS Coronavirus 2 (SARS-CoV2): Mutation in the Receptor Binding Domain of Spike Gene

  • Sinae Kim;Jong Ho Lee;Siyoung Lee;Saerok Shim;Tam T. Nguyen;Jihyeong Hwang;Heijun Kim;Yeo-Ok Choi;Jaewoo Hong;Suyoung Bae;Hyunjhung Jhun;Hokee Yum;Youngmin Lee;Edward D. Chan;Liping Yu;Tania Azam;Yong-Dae Kim;Su Cheong Yeom;Kwang Ha Yoo;Lin-Woo Kang;Kyeong-Cheol Shin;Soohyun Kim
    • IMMUNE NETWORK
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    • v.20 no.5
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    • pp.41.1-41.11
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    • 2020
  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) is a positive-sense single-stranded RNA (+ssRNA) that causes coronavirus disease 2019 (COVID-19). The viral genome encodes twelve genes for viral replication and infection. The third open reading frame is the spike (S) gene that encodes for the spike glycoprotein interacting with specific cell surface receptor - angiotensin converting enzyme 2 (ACE2) - on the host cell membrane. Most recent studies identified a single point mutation in S gene. A single point mutation in S gene leading to an amino acid substitution at codon 614 from an aspartic acid 614 into glycine (D614G) resulted in greater infectivity compared to the wild type SARS-CoV2. We were interested in investigating the mutation region of S gene of SARS-CoV2 from Korean COVID-19 patients. New mutation sites were found in the critical receptor binding domain (RBD) of S gene, which is adjacent to the aforementioned D614G mutation residue. This specific sequence data demonstrated the active progression of SARS-CoV2 by mutations in the RBD of S gene. The sequence information of new mutations is critical to the development of recombinant SARS-CoV2 spike antigens, which may be required to improve and advance the strategy against a wide range of possible SARS-CoV2 mutations.

Identification of differentially displayed genes from a soybean (Giycine max) cultivar resistant to a strain of Pseudomonas aeroginosa

  • Cha, Hyeon-Wook;Kang, Sang-Gu;Chang, Moo-Ung;Park, Euiho
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.72.2-73
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    • 2003
  • We found a soybean (Glycine max) cultivar 561 that was strongly resistant to a virulent bacterial strain of a Pseudomonas spp. Further identification revealed that the Pseudomonas spp. was a strain of Pseudomonas aeruginosa. Furthermore we identified specific genes involved in the resistance of soybean 561 and analyzed the pattern of gene expression against the Pseudomonas infection using differential-display reverse transcription PCR (DDRT-PCR). More than 126 cDNA fragments representing mRNAs were induced within 48 hours of bacteria inoculation. Among them, 28 cDNA fragments were cloned and sequenced. Twelve differentially displayed clones with open reading frames had unknown functions. Sixteen selected cDNA clones were homologous to known genes in the other organisms. Some of the identified cDNAs were pathogenesis-related genes (PR genes) and PR-like genes. These cDNAs included a putative calmodulin-binding protein, an endo-1,3-1,4-b-D-glucanase, a b-1,3-endoglucanase, a b-1,3-exoglucanase, a phytochelatin synthetase-like gene, a thiol pretense, a cycloartenol synthase, and a putative receptor-like sorineithreonine protein kinase. Among them, we found that four genes were putative pathogenesis-related genes (PR) induced significantly by the p. aeruginosa infection. These included a calmodulin-binding protein gene, a b-1,3-endoglucanase gene, a receptor-like sorine/threonine protein kinase gene, and pS321 (unknown function). These results suggest that the differentially expressed genes may mediate the strong resistance of soybean 561 to Pseudomonas aeruoginosa.

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Transcriptomic analysis of the liver in aged laying hens with different intensity of brown eggshell color

  • Han, Gi Ppeum;Kim, Jun-Mo;Kang, Hwan Ku;Kil, Dong Yong
    • Animal Bioscience
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    • v.34 no.5
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    • pp.811-823
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    • 2021
  • Objective: Eggshell color is an important indicator of egg quality for consumers, especially for brown eggs. Various factors related to laying hens and their environment affect brown eggshell coloration. However, there have been no studies investigating hepatic functions of laying hens with variable intensity of brown eggshell color. Therefore, this study was aimed to identify potential factors affecting brown eggshell coloration in aged laying hens at the hepatic transcriptomic level. Methods: Five hundred 92-wk-old Hy-line Brown laying hens were screened to select laying hens with different intensity of brown eggshell color based on eggshell color fans. Based on eggshell color scores, hens with dark brown eggshells (DBE; eggshell color fan score = 14.8) and hens with light brown eggshells (LBE; eggshell color fan score = 9.7) were finally selected for the liver sampling. We performed RNA-seq analysis using the liver samples through the paired-end sequencing libraries. Differentially expressed genes (DEGs) profiling was carried out to identify their biological meaning by bioinformatics. Results: A total of 290 DEGs were identified with 196 being up-regulated and 94 being down-regulated in DBE groups as compared to LBE groups. The Kyoto encyclopedia of genes and genomes (KEGG) analysis revealed that these DEGs belong to several biological pathways including herpes simplex infection (toll-like receptor 3 [TLR3], cyclin-dependent kinase 1, etc.) and influenza A (TLR3, radical S-adenosyl methionine domain containing 2, myxovirus [influenza virus] resistance 1, etc.). Genes related to stress response (ceremide kinase like) and nutrient metabolism (phosphoenolpyruvate carboxy-kinase 1, methylmalonic aciduria [cobalamin deficiency] cblB type, glycine receptor alpha 2, solute carrier family 7 member 11, etc.) were also identified to be differentially expressed. Conclusion: The current results provide new insights regarding hepatic molecular functions related to different intensity of brown eggshell color in aged laying hens. These insights will contribute to future studies aiming to optimize brown eggshell coloration in aged laying hens.

A MOLECULAR BIOLOGICAL STUDY ON THE EXPRESSION PATTERN AND FUNCTIONAL PROTEIN STRUCTURES OF PROLINE-RICH PROTEINS IN HUMAN SALIVARY GLANDS (사람의 타액선에서 proline-rich protein의 발현양상과 기능적 단백 구조에 대한 분자생물학적 연구)

  • Joo, Jae-Yong;Lee, Suk-Keun;Park, Young-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.1
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    • pp.31-41
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    • 2002
  • Proline-rich proteins (PRPs) are major components of human saliva. In order to know the biological roles of PRPs, we explored the expression pattern and functional protein structures of PRPs by the immunohistochemical and various molecular biological methods. Polyclonal antibody against human gPRP was generated from rabbit by the injection of oral exfoliated cells specially treated by urea and SDS buffer. The PRPs began to be expressed both in the acinar cells and ductal cells from the EIDS (Early Intermediate Developmental Stage) of fetal salivary glands and became intense in the salivary epithelium in the LDS (Late Developmental Stage) and adult salivary glands. The polyclonal antibody against the gPRP showed the cross-reactivity with aPRP and bPRP, these results were relevant to the high homology among subtypes of PRP. However, the simulated protein structures of PRPs showed the characteristic repetitive whorling domains except the N-terminal signal peptide. The whorling domains were also contained the multiple amino acids of glutamine and glycine, which may provide the receptor binding or cross-linking sites of PRPs.

Preparation and Characteristics of Bread by Medicinal Herb Composites with Cognitive Function (인지기능활성을 가진 생약복합물을 이용한 빵의 제조 및 특성)

  • Kang, Jin-Soon;Kang, Shin-Kwon;Kim, Hee-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.9
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    • pp.1131-1138
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    • 2009
  • In this study, the breads with medicinal herbs (MH) composites showing cognitive function were prepared and their characteristics were examined. Fifteen kinds of medicinal herbs were extracted with hot water and divided into 3 groups (MH-1, MH-2, MH-3) based on their contents. All groups showed cognitive function in terms of promoting muscarinic receptor, NMDA receptor and acetylcholinesterase inhibition. In the preparation of breads containing MH extracts of various contents (0, 10, 20, 30, and 50%), there were no significant difference among loaf weight of all groups, but loaf volume of all groups were significantly (p<0.05) decreased with increasing content of MH extracts compared to the control group. The "a" and "b" values of bread crumb increased with the content of MH extracts while "L" value decreased, but these values of bread crust were similar to the control group. The most improvements in hardness, fracturability, cohesiveness, gumminess and chewiness of bread were noticed by the addition of 10$\sim$20% MH extracts, while adhesiveness and springiness of bread were the most by the 30% addition ones. Through the sensory evaluation, it was revealed that appearance, mouth feeling, taste, overall preference and crumb texture of bread were not significantly different (p<0.05) between control and those with the 10% addition ones. Therefore the 10% addition ones, which got similar scores to control, will make the most desirable product.

Genetic Characterization of H7-subtype Avian Influenza Viruses (H7 아형 조류인플루엔자 바이러스의 유전자 특성)

  • Yeo, Jiin;Kwon, Hyuk-Moo;Sung, Haan-Woo
    • Korean Journal of Poultry Science
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    • v.46 no.3
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    • pp.173-183
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    • 2019
  • Based on their virulence, the avian influenza viruses (AIVs) are classified into two pathotypes: low pathogenic avian influenza (LPAI) virus and highly pathogenic avian influenza (HPAI) virus. Among the 16 HA subtypes of AIV, only the H5 and H7 subtypes are classified as HPAI. Some AIVs, including H5 and H7 viruses, can infect humans directly. Six H7 subtype isolates from wild birds of the H7N7 (n=4) and H7N1 (n=2) subtypes were characterized in this study. Phylogenetic analysis showed that eight viral genes (HA, NA, PB2, PB1, PA, NP, M, and NS) of the H7 isolates clustered in the Eurasian lineage, the genetic diversity of which is indicated by its division into several sublineages. The Korean H7 isolates had two motifs, PEIPKGR and PELPKGR, at the HA cleavage site, which have been associated with LPAI viruses. Six H7 isolates encoded glutamine (Q) and glycine (G) at positions 226 (H3 numbering) and 228 of HA, suggesting avian-type receptor-binding specificity. None of the Korean H7 isolates had the amino acid substitutions E627K in PB2 and I368V in PB1, which are critical for efficient replication in human cells. The Korean H7 isolates showed no deletions in the NA stalk region and in NS. These results suggest that the Korean H7 isolates from wild birds are different from the H7N9 influenza viruses isolated in China in 2013, which are capable of infecting humans.

Isolation and electrical characterization of the rat spinal dorsal horn neurons

  • Han, Seong-Kyu;Lee, Mun-Han;Ryu, Pan-Dong
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.175-175
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    • 1996
  • The spinal dorsal horn is the area where primary afferent fibers terminate and cutaneous sensory information is Processed. A number of putative neurotransmitter substances, including excitatory and inhibitory amino acids and peptides, are present in this region and sites and cellular mechanisms of their actions have been a target of numerous studies. In this study, single neurons were acutely isolated and the properties of whole cell current and responses to excitatory and inhibitory neurotransmitters were studied by the patch clamp method. Young rats (7-14 days) were anesthetized with diethyl-ether, and the lumbar spinal cord was excised and cut transversely at a thickness of 30$\mu\textrm{m}$ by Vibroslicer. The treatment of spinal slices with low concentration of proteases (pronase and thermolysin 0.75 mg/$m\ell$) and mechanical dissociation yielded isolated neurons with near intact morphology. Multipolar, ellipsoidal and bipolar, and pyramidal cells were shown. By applying step voltage pulses to neurons held at -70 mV, two types of inward currents and one outward currents observed. The fast activating and inactivating inward current was the Na$\^$+/ current because of its fast kinetics and blocking by 0.5${\mu}$M TTX, a specific blocker of Na$\^$+/ channel. The second type of inward currents were sustained. Based on their kinetics and current-voltage relations, it was likely that the second type of inward current was the voltage-dependent Ca$\^$2+/ current. In the presence of TTX, the steady-state currents mainly represented outward K$\^$+/ current which looked like the delayed rectifier K$\^$+/ current. In addition, the membrane currents produced by agonist of excitatory amino acid (EAA) receptor and the endogenous transmitter candidate L-glutamate were recorded in isolated whole-cell voltage clamped neurons as well as responses to inhibitory amino acids (${\gamma}$-amino butyric acid, glycine). Drugs were applied by a method that allows complete exchange of the solution within 1 sec; an infinite number of solutions can be applied to a single cell.

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Influence of Strychnine on Catecholamine Release Evoked by Activation of Cholinergic Receptors from the Perfused Rat Adrenal Gland

  • Yu, Byung-Sik;Kim, Byeong-Cheol;Oh, Song-Hoon;Kim, Il-Sik;Lee, Bang-Hun;Cho, Seong-Ho;Lim, Dong-Yoon
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.3
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    • pp.243-251
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    • 2001
  • The present study was attempted to investigate the effect of strychnine on catecholamine (CA) secretion evoked by ACh, high $K^+,$ DMPP and McN-A-343 from the isolated perfused rat adrenal gland. The perfusion of strychnine $(10^{-4}\;M)$ into an adrenal vein for 20 min produced great inhibition in CA secretory responses evoked by ACh $(5.32{\times}10^{-3}\;M),$ DMPP $(10^{-4}\;M\;for\;2\;min)$ and McN-A-343 $(10^{-4}\;M\;for\;2\;min),$ but did not alter CA secretion by high $K^+\;(5.6{\times}10^{-2}\;M).$ Strychnine itself did also fail to affect basal catecholamine output. Furthermore, in adrenal glands preloaded simultaneously with strychnine $(10^{-4}\;M)$ and glycine (an agonist of glycinergic receptor, $10^{-4}\;M),$ CA secretory responses evoked by ACh, DMPP and McN-A-343 were considerably recovered to some extent when compared with those evoked by treatment with strychnine only. However, CA secretion by high $K^+\;(5.6{\times}10^{-2}\;M)$ was not affected. Taken together, these results demonstrate that strychnine inhibits greatly the CA secretory responses evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors, but does not affect that by membrane depolarization. It is suggested that strychnine-sensitive glycinergic receptors are localized in rat adrenal medullary chromaffin cells.

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Combined transcriptome and proteome analyses reveal differences in the longissimus dorsi muscle between Kazakh cattle and Xinjiang brown cattle

  • Yan, XiangMin;Wang, Jia;Li, Hongbo;Gao, Liang;Geng, Juan;Ma, Zhen;Liu, Jianming;Zhang, Jinshan;Xie, Penggui;Chen, Lei
    • Animal Bioscience
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    • v.34 no.9
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    • pp.1439-1450
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    • 2021
  • Objective: With the rapid development of proteomics sequencing and RNA sequencing technology, multi-omics analysis has become a current research hotspot. Our previous study indicated that Xinjiang brown cattle have better meat quality than Kazakh cattle. In this study, Xinjiang brown cattle and Kazakh cattle were used as the research objects. Methods: Proteome sequencing and RNA sequencing technology were used to analyze the proteome and transcriptome of the longissimus dorsi muscle of the two breeds of adult steers (n = 3). Results: In this project, 22,677 transcripts and 1,874 proteins were identified through quantitative analysis of the transcriptome and proteome. By comparing the identified transcriptome and proteome, we found that 1,737 genes were identified at both the transcriptome and proteome levels. The results of the study revealed 12 differentially expressed genes and proteins: troponin I1, crystallin alpha B, cysteine, and glycine rich protein 3, phosphotriesterase-related, myosin-binding protein H, glutathione s-transferase mu 3, myosin light chain 3, nidogen 2, dihydropyrimidinase like 2, glutamate-oxaloacetic transaminase 1, receptor accessory protein 5, and aspartoacylase. We performed functional enrichment of these differentially expressed genes and proteins. The Kyoto encyclopedia of genes and genomes results showed that these differentially expressed genes and proteins are enriched in the fatty acid degradation and histidine metabolism signaling pathways. We performed parallel reaction monitoring (PRM) verification of the differentially expressed proteins, and the PRM results were consistent with the sequencing results. Conclusion: Our study provided and identified the differentially expressed genes and proteins. In addition, identifying functional genes and proteins with important breeding value will provide genetic resources and technical support for the breeding and industrialization of new genetically modified beef cattle breeds.

Neuroprotective Effects of Ginsenoside Rg3 against 24-OH-cholesterol-induced Cytotoxicity in Cortical Neurons

  • Roh, Yoon-Seok;Kim, Hyoung-Bae;Kang, Chang-Won;Kim, Bum-Seok;Nah, Seung-Yeol;Kim, Jong-Hoon
    • Journal of Ginseng Research
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    • v.34 no.3
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    • pp.246-253
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    • 2010
  • Ginsenoside $Rg_3$ ($Rg_3$), one of the active ingredients in Panax ginseng, attenuates NMDA receptor-mediated currents in vitro and antagonizes NMDA receptors through a glycine modulatory site in rat cultured hippocampal neurons. In the present study, we examined the neuroprotective effects of $Rg_3$ on 24-hydroxycholesterol (24-OH-chol)-induced cytotoxicity in vitro. The results showed that $Rg_3$ treatment significantly and dose-dependently inhibited 24-OH-chol-induced cell death in rat cultured cortical neurons, with an $IC_{50}$ value of $28.7{\pm}7.5\;{\mu}m$. Furthermore, the $Rg_3$ treatment not only significantly reduced DNA damage, but also dose-dependently attenuated 24-OH-chol-induced caspase-3 activity. To study the mechanisms underlying the in vitro neuroprotective effects of $Rg_3$ against 25-OH-chol-induced cytotoxicity, we also examined the effect of $Rg_3$ on intracellular $Ca^{2+}$ elevations in cultured neurons and found that $Rg_3$ treatment dose-dependently inhibited increases in intracellular $Ca^{2+}$, with an $IC_{50}$ value of $40.37{\pm}12.88\;{\mu}m$. Additionally, $Rg_3$ treatment dose-dependently inhibited apoptosis with an $IC_{50}$ of $47.3{\pm}14.2\;{\mu}m$. Finally, after confirming the protective effect of $Rg_3$ using a terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assay, we found that $Rg_3$ is an active component in ginseng-mediated neuroprotection. These results collectively indicate that $Rg_3$-induced neuroprotection against 24-OH-chol in rat cortical neurons might be achieved via inhibition of a 24-OH-chol-mediated $Ca^{2+}$ channel. This is the first report to employ cortical neurons to study the neuroprotective effects of $Rg_3$ against 24-OH-chol. In conclusion, $Rg_3$ was effective for protecting cells against 24-OH-chol-induced cytotoxicity in rat cortical neurons. This protective ability makes $Rg_3$ a promising agent in pathologies implicating neurodegeneration such as apoptosis or neuronal cell death.