• 제목/요약/키워드: Cell mass

검색결과 2,608건 처리시간 0.032초

NADP+-Dependent Dehydrogenase SCO3486 and Cycloisomerase SCO3480: Key Enzymes for 3,6-Anhydro-ʟ-Galactose Catabolism in Streptomyces coelicolor A3(2)

  • Tsevelkhorloo, Maral;Kim, Sang Hoon;Kang, Dae-Kyung;Lee, Chang-Ro;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • 제31권5호
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    • pp.756-763
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    • 2021
  • Agarose is a linear polysaccharide composed of ᴅ-galactose and 3,6-anhydro-ʟ-galactose (AHG). It is a major component of the red algal cell wall and is gaining attention as an abundant marine biomass. However, the inability to ferment AHG is considered an obstacle in the large-scale use of agarose and could be addressed by understanding AHG catabolism in agarolytic microorganisms. Since AHG catabolism was uniquely confirmed in Vibrio sp. EJY3, a gram-negative marine bacterial species, we investigated AHG metabolism in Streptomyces coelicolor A3(2), an agarolytic gram-positive soil bacterium. Based on genomic data, the SCO3486 protein (492 amino acids) and the SCO3480 protein (361 amino acids) of S. coelicolor A3(2) showed identity with H2IFE7.1 (40% identity) encoding AHG dehydrogenase and H2IFX0.1 (42% identity) encoding 3,6-anhydro-ʟ-galactonate cycloisomerase, respectively, which are involved in the initial catabolism of AHG in Vibrio sp. EJY3. Thin layer chromatography and mass spectrometry of the bioconversion products catalyzed by recombinant SCO3486 and SCO3480 proteins, revealed that SCO3486 is an AHG dehydrogenase that oxidizes AHG to 3,6-anhydro-ʟ-galactonate, and SCO3480 is a 3,6-anhydro-ʟ-galactonate cycloisomerase that converts 3,6-anhydro-ʟ-galactonate to 2-keto-3-deoxygalactonate. SCO3486 showed maximum activity at pH 6.0 at 50℃, increased activity in the presence of iron ions, and activity against various aldehyde substrates, which is quite distinct from AHG-specific H2IFE7.1 in Vibrio sp. EJY3. Therefore, the catabolic pathway of AHG seems to be similar in most agar-degrading microorganisms, but the enzymes involved appear to be very diverse.

Characterization of a Thermostable Lichenase from Bacillus subtilis B110 and Its Effects on β-Glucan Hydrolysis

  • Huang, Zhen;Ni, Guorong;Wang, Fei;Zhao, Xiaoyan;Chen, Yunda;Zhang, Lixia;Qu, Mingren
    • Journal of Microbiology and Biotechnology
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    • 제32권4호
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    • pp.484-492
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    • 2022
  • Lichenase is an enzyme mainly implicated in the degradation of polysaccharides in the cell walls of grains. Emerging evidence shows that a highly efficient expression of a thermostable recombinant lichenase holds considerable promise for application in the beer-brewing and animal feed industries. Herein, we cloned a lichenase gene (CelA203) from Bacillus subtilis B110 and expressed it in E. coli. This gene contains an ORF of 729 bp, encoding a protein with 242 amino acids and a calculated molecular mass of 27.3 kDa. According to the zymogram results, purified CelA203 existed in two forms, a monomer, and a tetramer, but only the tetramer had potent enzymatic activity. CelA203 remained stable over a broad pH and temperature range and retained 40% activity at 70℃ for 1 h. The Km and Vmax of CelA203 towards barley β-glucan and lichenan were 3.98 mg/ml, 1017.17 U/mg, and 2.78 mg/ml, 198.24 U/mg, respectively. Furthermore, trisaccharide and tetrasaccharide were the main products obtained from CelA203-mediated hydrolysis of deactivated oat bran. These findings demonstrate a promising role for CelA203 in the production of oligosaccharides in animal feed and brewing industries.

Gintonin-enriched fraction improves sarcopenia by maintaining immune homeostasis in 20- to 24-month-old C57BL/6J mice

  • Oh, Hyun-Ji;Jin, Heegu;Nah, Seung-Yeol;Lee, Boo-Yong
    • Journal of Ginseng Research
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    • 제45권6호
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    • pp.744-753
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    • 2021
  • Background: Gintonin-enriched fraction (GEF) is a new non-saponin component glycolipoprotein isolated from ginseng root. This study examined the effect of GEF on age-related sarcopenia in old C57BL/6J mice. Methods: Young (3-6 months) and old (20-24 months) C57BL/6J mice received oral GEF (50 mg/kg/day or 150 mg/kg/day) daily for 5 weeks. During the oral administration period, body weight and grip strength were measured weekly. After sacrifice, muscles from the hindlimb were excised and used for hematoxylin and eosin staining and western blotting to determine the effects of GEF on sarcopenia. The thymus was photographed to compare size, and flow cytometry was performed to examine the effect of GEF on immune homeostasis in the thymus and spleen. Blood samples were collected, and the concentrations of pro-inflammatory cytokines and IGF-1 were measured. Results: GEF caused a significant increase in muscle strength, mass, and fiber size in old mice. GEF restored age-related disruption of immune homeostasis by maintaining T cell compartments and regulating inflammatory biomarkers. Thus, GEF reduced common low-grade chronic inflammatory parameters, which are the main cause of muscle loss. Conclusion: GEF maintained immune homeostasis and inhibited markers of chronic inflammation, resulting in anti-sarcopenia effects in aged C57BL/6J mice. Thus, GEF is a potential therapeutic agent that slows sarcopenia in the elderly.

Novel Antibacterial, Cytotoxic and Catalytic Activities of Silver Nanoparticles Synthesized from Acidophilic Actinobacterial SL19 with Evidence for Protein as Coating Biomolecule

  • Wypij, Magdalena;Ostrowski, Maciej;Piska, Kamil;Wojcik-Pszczola, Katarzyna;Pekala, Elzbieta;Rai, Mahendra;Golinska, Patrycja
    • Journal of Microbiology and Biotechnology
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    • 제32권9호
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    • pp.1195-1208
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    • 2022
  • Silver nanoparticles (AgNPs) have potential applications in medicine, photocatalysis, agriculture, and cosmetic fields due to their unique physicochemical properties and strong antimicrobial activity. Here, AgNPs were synthesized using actinobacterial SL19 strain, isolated from acidic forest soil in Poland, and confirmed by UV-vis and FTIR spectroscopy, TEM, and zeta potential analysis. The AgNPs were polydispersed, stable, spherical, and small, with an average size of 23 nm. The FTIR study revealed the presence of bonds characteristic of proteins that cover nanoparticles. These proteins were then studied by using liquid chromatography with tandem mass spectrometry (LC-MS/MS) and identified with the highest similarity to hypothetical protein and porin with molecular masses equal to 41 and 38 kDa, respectively. Our AgNPs exhibited remarkable antibacterial activity against Escherichia coli and Pseudomonas aeruginosa. The combined, synergistic action of these synthesized AgNPs with commercial antibiotics (ampicillin, kanamycin, streptomycin, and tetracycline) enabled dose reductions in both components and increased their antimicrobial efficacy, especially in the case of streptomycin and tetracycline. Furthermore, the in vitro activity of the AgNPs on human cancer cell lines (MCF-7, A375, A549, and HepG2) showed cancer-specific sensitivity, while the genotoxic activity was evaluated by Ames assay, which revealed a lack of mutagenicity on the part of nanoparticles in Salmonella Typhimurium TA98 strain. We also studied the impact of the AgNPs on the catalytic and photocatalytic degradation of methyl orange (MO). The decomposition of MO was observed by a decrease in intensity of absorbance within time. The results of our study proved the easy, fast, and efficient synthesis of AgNPs using acidophilic actinomycete SL19 strain and demonstrated the remarkable potential of these AgNPs as anticancer and antibacterial agents. However, the properties and activity of such particles can vary by biosynthesized batch.

Utilization of Supplemental Regional Anesthesia in Lobectomy for Lung Cancer in the United States: A Retrospective Study

  • Alwatari, Yahya;Vudatha, Vignesh;Scheese, Daniel;Rustom, Salem;Ayalew, Dawit;Sevdalis, Athanasios E.;Julliard, Walker;Shah, Rachit D.
    • Journal of Chest Surgery
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    • 제55권3호
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    • pp.225-232
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    • 2022
  • Background: Pulmonary lobectomy is the standard of care for the treatment of early-stage non-small cell lung cancer. This study investigated the rate of utilization of supplemental anesthesia in patients undergoing video-assisted thoracoscopic surgery (VATS) or open lobectomy using a national database and assessed the effect of regional block (RB) on postoperative outcomes. Methods: Patients who underwent lobectomy for lung cancer between 2014-2019 were identified in the American College of Surgeons National Surgical Quality Improvement Program. The patients' primary mode of anesthesia and supplemental anesthesia were recorded. Preoperative characteristics and postoperative outcomes were compared between 2 surgical groups: those who underwent general anesthesia (GA) alone versus GA with RB. Multivariable regression analyses were performed on the outcomes of interest. Results: In total, 13,578 patients met the study criteria, with 87% undergoing GA and the remaining 13% receiving GA and RB. The use of neuraxial anesthesia decreased over the years, while RB use increased up to 20% in 2019. Age, body mass index, and preoperative comorbidities were comparable between groups. Patients who underwent VATS were more likely to receive RB than those who underwent thoracotomy. RB was most often utilized by thoracic surgeons. An adjusted analysis showed that RB use was associated with shorter hospital stays and a reduced likelihood of prolonged length of stay, but a higher rate of surgical site infections (SSIs). Conclusion: In a large surgical database, there was underutilization of supplemental anesthesia in patients undergoing lobectomy for lung cancer. RB utilization was associated with a shorter length of hospital stay and an increase in SSI incidence.

통증을 동반한 신경총병증에서 언제 신경림프종증을 고려해야 하는가? (When We Consider Neurolymphomatosis in Patient with Lumbosacral Plexopathy with an Extreme Leg Pain?)

  • 안준영;석현;김상현;김현정;조연희;오백민;이승열
    • Clinical Pain
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    • 제20권1호
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    • pp.53-57
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    • 2021
  • We report a case of neurolymphomatosis of lumbosacral plexus. A 63-year-old man, who had no past history except for diabetes mellitus, complained of severe pain and weakness on left lower extremity. Idiopathic lumbosacral plexopathy was diagnosed by electromyography. There were no abnormal findings except for FDG-PET/CT and MRI. They showed high uptake and thickening lesion in sciatic nerve and sacral plexus. However, about 7 months later, mass like lesion in left thigh was detected by FDG-PET/CT and MRI. Also, multiple hypermetabolic lesions were found in brain. Through brain biopsy, diffuse large B-cell lymphoma was confirmed. When a patient with idiopathic lumbosacral plexopathy complains of severe pain, it is necessary to consider FDG-PET/CT and MRI to differentiate neurolymphomatosis, even in patients who have no past history of lymphoma before. Especially, if FDG-PET/CT and MRI show sciatic and/or lumbosacral plexus lesion, neurolymphomatosis of lumbosacral plexus should be considered.

Identification, Characterization, and Efficacy Evaluation of Bacillus velezensis for Shot-Hole Disease Biocontrol in Flowering Cherry

  • Han, Viet-Cuong;Yu, Nan Hee;Yoon, Hyeokjun;Ahn, Neung-Ho;Son, Youn Kyoung;Lee, Byoung-Hee;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • 제38권2호
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    • pp.115-130
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    • 2022
  • Though information exists regarding the pathogenesis of the shot-hole disease (SH) in flowering cherry (FC), there has been a lack of research focusing on SH management. Therefore, here, we investigated the inhibitory activities of antagonistic bacteria against SH pathogens both in vitro and in vivo as well as their biochemical characteristics and bioactive compounds. Two biosurfactant-producing bacterial antagonists, identified as Bacillus velezensis strains JCK-1618 and JCK-1696, exhibited the best effects against the growth of both bacterial and fungal SH pathogens in vitro through their cell-free culture filtrates (CFCFs). These two strains also strongly inhibited the growth of the pathogens via the action of their antimicrobial diffusible compounds and antimicrobial volatile organic compounds (VOCs). Crude enzymes, solvent extracts, and biosurfactants of the two strains exhibited antimicrobial activities. Liquid chromatography/electrospray ionization time-of-flight mass spectrometric analysis of the partially purified active fractions revealed that the two antagonists produced three cyclic lipopeptides, including iturin A, fengycin A, and surfactin, and a polyketide, oxydifficidin. In a detached leaf assay, pre-treatment and co-treatment of FC leaves with the CFCFs led to a large reduction in the severity of the leaf spots caused by Epicoccum tobaicum and Bukholderia contaminans, respectively. In addition, the two antagonists produced indole-3-acetic acid, siderophore, and a series of hydrolytic enzymes, along with the formation of a substantial biofilm. To our knowledge, this is the first report of the antimicrobial activities of the diffusible compounds and VOCs of B. velezensis against the SH pathogens and their efficiency in the biocontrol of SH.

Biliary Obstruction Caused by Non-Hodgkin Lymphoma Involvement: A Case Report

  • Jae Hyun Lim;Huapyong Kang;Jung Hyun Jo;Hee Seung Lee;Jeong Youp Park;Seungmin Bang;Seung Woo Park;Si Young Song;Moon Jae Chung
    • Journal of Digestive Cancer Research
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    • 제6권1호
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    • pp.32-35
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    • 2018
  • Non-Hodgkin's lymphoma is known to be a rare and unusual cause of biliary obstruction. We report a case of biliary obstruction that a 25-year-old male showed icteric sclera and yellow discoloration of his skin caused by metastasis of non-Hodgkin lymphoma. Initial imaging & endoscopic work-up led us to an impression of either cholangiocarcinoma or IgG4-related disease, yet the pathological results weren't diagnostic. Through our thorough re-examination, we found a 5cm sized round, fixed, non-tender sternal mass, and additional imaging studies were suggestive of lymphoma, which was also consistent with the results of incisional chest wall biopsy. Biliary obstruction by lymphoma was successfully treated by endoscopic plastic stent insertion procedure and chemotherapy. Although it is widely accepted that lymphoma accounts for very few portion of malignant biliary obstruction, due to the fact that lymphoma and cholangiocarcinoma are often indistinguishable, careful diagnostic approach should be done.

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백산차(Ledum palustre var. diversipilosum) 종자의 형태적 특성 및 발아조건 분석 (Analysis on Morphological Characters and Germination Conditions of Seeds in Ledum palustre var. diversipilosum)

  • 최영민;조승주;이현지;김명자;성정원;윤정원
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2023년도 임시총회 및 춘계학술대회
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    • pp.23-23
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    • 2023
  • The Hairy labrador tea (Ledum palustre L. var. diversipilosum Nakai) is a evergreen small shrub, belonging to the Ericaceae and mainly distributed along the alpine areas of Hamgyeong-do in North Korea. Through seed physiological research on North Korean plants, we intend to obtain basic data for the development of mass propagation methods and use them for vegetation restoration. The internal and external morphological characters of the seed were observed using scanning electron microscopy or stereoscopy, and full seeds were selected through X-ray test. Seeds were cultured on a growth condition at 15/6℃ and 25/15℃ to examine the germination response according to temperature, and the number of germinated individuals was examined every 24 hours to calculate germination percentage and mean germination time. Low-temperature treatment (4℃) and hormone treatment (GA3) were performed to investigate the effect of dormancy breaking, and analysis of variance (ANOVA) was performed on the experimental results using SAS 9.4. The seeds of the Hairy labrador tea were elliptical-narrow in shape, and the epidermal cell wall pattern was reticulated. The final germination rate (FGP) was the highest at 58.9% under the condition of low temperature treatment at 25/15℃. The average number of days to germination (MGT) was the highest at 25.1 days without treatment at 15/6℃, and the average of the last germination days was confirmed to be about 13 days. As a result of low temperature treatment, it was confirmed that the final germination rate (FGP) increased by 43.3%. The significance of the final germination rate (FGP) was verified according to the pretreatment at 25/15℃.

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The characteristics of bovine satellite cells with highly scored genomic estimated breeding value

  • Jae Ho Han;Ji Suk Yu;Do Hyun Kim;Hyun Woo Choi
    • 한국동물생명공학회지
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    • 제38권3호
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    • pp.177-187
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    • 2023
  • Background: The grading of Hanwoo (Korean native cattle) is based on four economic traits, and efforts have been continuously made to improve the genetic traits associated with these traits. There is a technology to predict the expected grade based on the 4 economic genetic SNP characteristics of Korean cattle calves using single nucleotide polymorphism (SNP) technology. Selection of highly proliferative, self-renewing, and differentiating satellite cells from cattle is a key technology in the cultured meat industry. Methods: We selected the Hanwoo with high and low-scored of genomic estimated breeding value (GEBV) by using the Hanwoo 50K SNP bead chip. We then isolated the bovine satellite cells from the chuck mass. We then conducted comparative analyses of cell proliferation, immunocytochemistry, qRT-PCR at short- and long-term culture. We also analyzed the differentiation capability at short term culture. Results: Our result showed that the proliferation was significantly high at High scored GEBV (Hs-GEBV) compared to Low scored GEBV (Ls-GEBV) at short- and long-term culture. The expression levels of Pax3 were significantly higher in Hs-GEBV bovine satellite cells at long-term culture. However, there were no significant differences in the expression levels of Pax7 between Hs- and Ls-GEBV bovine satellite cells at short- and long- term culture. The expression levels of MyoG and MyHC were significantly high at Ls-GEBV bovine satellite cells. Conclusions: Our results indicated that selection of bovine satellite cells by Hanwoo 50K SNP bead chip could be effective selection methods for massive producing of satellite cells.