• International Journal of Stem Cells
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• v.16 no.2
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• pp.191-201
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• 2023

Background and Objectives: O-cyclic phytosphingosine-1-phosphate (cP1P) is a synthetic chemical and has a structure like sphingosine-1-phosphate (S1P). S1P is known to promote cell migration, invasion, proliferation, and anti-apoptosis through hippocampal signals. However, S1P mediated cellular-, molecular mechanism is still remained in the lung. Acute lung injury (ALI) and its severe form acute respiratory distress syndrome (ARDS) are characterized by excessive immune response, increased vascular permeability, alveolar-peritoneal barrier collapse, and edema. In this study, we determined whether cP1P primed human dermal derived mesenchymal stem cells (hdMSCs) ameliorate lung injury and its therapeutic pathway in ALI mice. Methods and Results: cP1P treatment significantly stimulated MSC migration and invasion ability. In cytokine array, secretion of vascular-related factors was increased in cP1P primed hdMSCs (hdMSC^cP1P), and cP1P treatment induced inhibition of Lats while increased phosphorylation of Yap. We next determined whether hdMSC^cP1P reduce inflammatory response in LPS exposed mice. hdMSC^cP1P further decreased infiltration of macrophage and neutrophil, and release of TNF-α, IL-1β, and IL-6 were reduced rather than naïve hdMSC treatment. In addition, phosphorylation of STAT1 and expression of iNOS were significantly decreased in the lungs of MSC^cP1P treated mice. Conclusions: Taken together, these data suggest that cP1P treatment enhances hdMSC migration in regulation of Hippo signaling and MSC^cP1P provide a therapeutic potential for ALI/ARDS treatment.

• Journal of Food Hygiene and Safety
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• v.39 no.3
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• pp.181-190
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• 2024

Bovine mastitis-associated Escherichia coli (BMEC) is considered the main causative agent of significant financial losses in the dairy industry worldwide, as it alters both the quantity and quality of milk produced and increases the rate of culling. This creates a variety of challenges for researchers, veterinarians, and farmers in understanding and determining the most effective therapies and diagnostic techniques. Subclinical mastitis is particularly concerning, as infected bovines exhibit no obvious symptoms and continue to secrete apparently normal milk over an extended period, allowing the causative pathogen, E. coli, to spread within the herd. For effective prevention, understanding the pathogenesis of mastitis through three stages invasion, infection, and inflammation is essential. To date, no clear correlation has been found between virulence factors and pathogenicity contributing to the clinical severity of BMEC. Multidrug-resistant E. coli and the evolution of novel resistance mechanisms have become concerns owing to the extensive use of antibiotics to treat mastitis. Therefore, it is vital to explore alternative controls to enhance the efficacy of BMEC treatment. Over the past 30 years, various genetic typing techniques have been used to examine the subspecies-level epidemiology of bovine mastitis. These studies have advanced our understanding of the origin, transmission pathway, population structure, and evolutionary relatedness of BMEC strains. In this review we provide an overview of BMEC, including insights into its etiology, genetic relationship, pathogenesis, and management of the disease, as well as new therapy options.

• Journal of Electrochemical Science and Technology
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• v.15 no.4
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• pp.466-475
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• 2024

This research used solid waste from sugarcane production, named bagasse, as raw material for a functional carbon electrode. The bagasse was carbonized to produce carbon powder and, following activation with water vapor at 700℃. The activated carbon was doped with N and S to improve its electrochemical properties by treating it with thiourea precursor and heating it at 850℃ under nitrogen flow to produce N/S-doped carbon (NSCE). The produced carbon was then characterized to understand the specific diffraction pattern, molecular vibrations, and surface morphology. The result found that the NSCE showed two broad diffraction peaks at 23° and 43°, corresponding to [002] and [100] crystal planes following JCPDS75-1621. FTIR spectra showed some O-H, C-H, C-O, and C=C peaks. Peaks of C=N, C-N, and S-H demonstrate the presence of N/S within the NSCE. Raman analysis revealed that N/S doping caused structure defects within the single C6 layer networks, providing carbon vacancies (V_C^••••) because of C replacement by N (N'_C) and S (S"_C). Meanwhile, XPS analysis showed N/S introduction to the C network by revealing peaks at 168.26 eV and 169.55 eV, corresponding to S2p_3/2 and S2p_1/2, and 171.95 eV corresponds to C-SO₃-C, indicating the presence of S within the thiol group attached to the carbon. Meanwhile, N1s are revealed at 402.4 eV and 405.5 eV, confirming pyrrolic nitrogen (N-5) and quaternary nitrogen (N-Q). The electrochemical analysis found that the reaction within the prepared-NSCE/NaClO₄/Na was reversible, with an onset potential of 0.1 V vs. Na/Na⁺, explaining the intercalation and deintercalation of sodium ions. The sodium battery full cell showed an excellent battery performance with an initial charging-discharging capacity of 720 mAh g^-1 and 570 mAh g^-1, respectively, at 0.2C. Meanwhile, a cycling test showed the average Coulombic efficiency of 84.4% and capacity retention of 57% after 50 cycles.

• Journal of Life Science
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• v.34 no.10
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• pp.682-690
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• 2024

This study aimed to produce medium-chain-length polyhydroxyalkanoates (mcl-PHA) using oil extracted from spent coffee grounds (SCG) as a carbon source. The highest oil yields, 14.40%(w/w) and 14.49%(w/w), were achieved with an extraction time of 2 hr and an SCG/n-hexane ratio of 15:1. The fatty acid composition of the extracted SCG oil (EOC) primarily consisted of linoleic acid (45.08%), palmitic acid (35.56%), oleic acid (8.62%), and stearic acid (7.62%). Pseudomonas sp. HY61, isolated from soil, was used to biosynthesize mcl-PHA composed of 3-hydroxyhexanoate, 3-hydroxyoctanoate, 3-hydroxydecanoate, and 3-hydroxydodecanoate using EOC. Under optimal flask conditions, the highest cell growth and mcl-PHA accumulation were observed with 5 g/l EOC, 72 hr, with NH₄Cl as the nitrogen source, at pH 7, 25℃, and a C/N ratio of 15:1. Batch fermentation resulted in a dry cell weight (DCW) of 0.92 g/l and mcl-PHA of 0.17 g/l after 72 hr. In contrast, fed-batch fermentation, with additional supplementation of EOC and NH₄Cl at 18, 30, and 42 hr, increased the DCW to 2.85 g/l and mcl-PHA accumulation to 0.67 g/l, approximately three times higher than batch fermentation. Gas chromatography confirmed that the mcl-PHA structure was consistent across both fermentation modes. The synthesized mcl-PHA had higher molecular weight and thermal decomposition temperatures compared to other mcl-PHA, suggesting its potential applications in packaging and biomedical fields. This study offers an economical approach for mcl-PHA production using waste EOC and Pseudomonas sp. HY61.

• Journal of the Korean Applied Science and Technology
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• v.33 no.2
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• pp.286-292
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• 2016

1, 2-Hexanediol galactoside (HD-gal) has been synthesized from 1, 2-hexanediol (HD), a cosmetic preservative, using recombinant Escherichia coli ${\beta}$-galactosidase (${\beta}$-gal) at the high lactose concentration (300 g/l). To confirm the molecular structure of synthesized HD-gal, NMR ($^1H$- and $^{13}C$-) spectroscopy and mass spectrometry of HD-gal were conducted. $^1H$ NMR spectrum of HD-gal showed multiple peaks corresponding to the galactocyl group, which is an evidence of galactocylation on HD. Downfield proton peaks at ${\delta}_H$ 4.44 ppm and multiple peaks from ${\delta}_H$3.96~3.58 ppm were indicative of galactocylation on HD. Up field proton peaks at ${\delta}_H$ 1.60~1.35 ppm and 0.92 ppm showed the presence of $CH_2$ and $CH_3$ protons of HD. $^{13}C$ NMR spectrum revealed the presence of 21 carbons suggestive of ${\alpha}$- and ${\beta}$-anomers of HD-gal. Among 12 carbon peaks from each anomers, the 3 peaks at dC 68.6, 60.9 and 13.2 ppm were assigned to be overlapped showing only 21 peaks out of total 24 peaks. The mass value (protonated HD-gal, m/z = 281.1601) from mass spectrometry analysis of HD-gal, and $^1H$ and $^{13}C$ NMR spectral data were in well agreement with the expecting structure of HD-gal. For further study, the minimum inhibitory concentrations (MICs) of HD-gal against bacteria will be investigated, and, in addition, cytotoxicity to human skin cells of HD-gal will be examined. It is expected that it will eventually be able to develop a new cosmetic preservative, which have low cytotoxicity against human skin cell and maintains antimicrobial effect.

• Applied Chemistry for Engineering
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• v.7 no.2
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• pp.252-260
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• 1996

In this article, we investigate into the structural changes of liposome to design its functional membranes by the synthesis of two types of liposomes, DPPC liposome and DTAB of hydrocarbon substance/DPPC liposome. The changes of membrane structures are evaluated by the CF fluorescent intensity measured above and below the phase transition temperature of the membrane, $t_c=41^{\circ}C$. CF fluorescent intensities are enhanced by the CF leakage from DPPC liposome at $45^{\circ}C$, while no changes are observed at $20^{\circ}C$. Under the same conditions, it is observed that the intensity enhanced by CF leakage from DPPC/DTAB liposome is larger than that of DPPC liposome alone, which suggests that DPPC/DTAB liposome has irregular arrangement. Under the presence of $Ca^{2+}$, Quin 2 fluorescent intensity in either DPPC liposome or DPPC/DTAB liposome is significantly increasing at $45^{\circ}C$, while almost none of the changes are observed at $20^{\circ}C$. The fluorescent intensity of DPPC liposome turns out to be larger than that of DPPC/DTAB liposome, which suggests that the DPPC/DTAB liposome is structurally more stable than the DPPC liposome. Additionally, when the analysis is done to observe changes in the shapes of membrane surfaces with ANS fluorescent, ANS fluorescent under DPPC or DPPC/DTAB liposome shows each of different appearances at $45^{\circ}C$ and $20^{\circ}C$ respectively. This result indicates that its respective membrane fluidity is changing above and below of the designated temperatures in phase transition. As to the magnitude of change of its membrane fluidity, DPPC liposome is much larger than DPPC/DTAB liposome. As far as the temperature in phase transition measured by DSC are concerned, it is $41^{\circ}C$ and $32^{\circ}C$ for DPPC and DPPC/DTAB liposome respectively, which suggests that DPPC/DTAB liposome has an irregular molecular arrangement in its structure. That is, it is summed up that DPPC/DTAB turns out to be structurally stable, even so, its structure is irregularly arranged.

• Proceedings of the KSAR Conference
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• 2000.10a
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• pp.10-12
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• 2000

Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

• Korean Journal of Plant Tissue Culture
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• v.24 no.1
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• pp.1-35
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• 1997

Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.

• Microbiology and Biotechnology Letters
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• v.37 no.3
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• pp.248-257
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• 2009

The inhibitors against Vibrio harveyi quorum sensing (QS) signaling were developed by modifying the molecular structure of the major signal, N-3-hydroxybutanoyl-L-homoserine lactone (3-OH-$C_4$-HSL). A series of structural derivatives, N-(3-hydroxysulfonyl)-L-homoserine lactones (HSHLs) were synthesized by the solid-phase organic synthesis method. The in vivo QS inhibition by these compounds was measured by a bioassay system using the V. harveyi bioluminescence, and all showed significant inhibitory effects. To analyze the interaction between these compounds and LuxN, a 3-OH-$C_4$-HSL receptor protein of V. harveyi, we tentatively determined the putative signal binding domain of LuxN based on the sequence homology with other acyl-HSL binding proteins, and predicted the partial 3-D structure of the putative signal binding domain of LuxN by using ORCHESTRA program, and further estimated the binding poses and energies (docking scores) of 3-OH-$C_4$-HSL and HSHLs within the domain. In comparison of the result from this modeling study with that of in vivo bioassay, we suggest that the in silica interpretation of the interaction between ligands and their receptor proteins can be a valuable way to develop better competitive inhibitors, especially in the case that the structural information of the protein is limited.

• Journal of the Korean Applied Science and Technology
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• v.37 no.5
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• pp.1323-1329
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• 2020

To characterize the molecular structure of PhE-gal synthesized using Escherichia coli 𝛽-gal, NMR (¹H- and ¹³C-) spectroscopy and mass spectrometry of PhE-gal were conducted. ¹H NMR spectrum of PhE-gal showed multiple peaks corresponding to the galactosyl group, which is an evidence of galactosylation on 2-phenylethanol (PhE). Downfield proton peaks at 𝛿_H 7.30~7.21 ppm showed the presence of aromatic protons of PhE as well as benzyl CH₂ protons at 𝛿_H 2.88 ppm. Up field proton peaks at 𝛿_H 4.31 ppm, 4.07 ppm and multiple peaks from 𝛿_H 3.86~3.38 ppm are indicative of galactocylation on PhE. ¹³C NMR spectrum revealed the presence of 12 carbons suggestive of PhE-gal. Among 12 carbon peaks from PhE-gal, the four peaks at 138.7, 129.0, 128.6 and 126.5 were assigned aromatic carbons in the phenyl ring. Three peaks at 129.0, 128.6 and 126.5 showed high intensities, indicating CH aromatic carbons. ¹³C NMR data of PhE-gal showed 6 monosaccharide peaks from galactose and 2 peaks from aliphatic chain of PhE, indicating that PhE-gal was galactosyl PhE. The mass value (sodium adduct ion of PhE-gal, m/z = 307.1181) from mass spectrometry analysis of PhE-gal, and ¹H and ¹³C NMR spectral data were in good agreement with the expecting structure of PhE-gal. We are expecting that through future study it will eventually be able to develop a new additive with low cytotoxicity.