• Title/Summary/Keyword: E-Isolation

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Bioactivity of Metabolites from Actinomycetes Isolates from Red Sea, Egypt

  • Osman, Mohamed E.;El-nasr, Amany A. Abo;Hussein, Hagar M;Hamed, Moaz M
    • Microbiology and Biotechnology Letters
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    • v.50 no.2
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    • pp.255-269
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    • 2022
  • Actinomycetes isolated from marine habitats represent a promising source of bioactive substances. Here, we report on the isolation, identification, productivity enhancement and application of the bioactive compounds of Streptomyces qinglanensis H4. Eighteen marine actinomycetes were isolated and tested for resistance to seven bacterial diseases. Using 16S rRNA sequencing analysis (GenBank accession number MW563772), the most powerful isolate was identified as S. qinglanensis. Although the strain produced active compound(s) against a number of Gram-negative and Gram-positive bacteria, it failed to inhibit pathogenic fungi. The obtained inhibition zones were 22.0 ± 1.5, 20.0 ± 1, 16.0 ± 1, 12.0 ± 1, 22.0 ± 1 and 24.0 ± 1 mm against Bacillus subtilis ATCC 6633, Escherichia coli ATCC 19404, Enterococcus faecalis ATCC 29212, Pseudomonas aeruginosa ATCC 9027, Candida albicans ATCC 10231 and Staphylococcus aureus ATCC6538, respectively. To maximize bioactive compound synthesis, the Plackett-Burman design was used. The productivity increased up to 0.93-fold, when S. qinglanensis was grown in optimized medium composed of: (g/l) starch 30; KNO3 0.5; K2HPO4 0.25; MgSO4 0.25; FeSO4·7H2O, 0.01; sea water concentration (%) 100; pH 8.0, and an incubation period of 9 days. Moreover, the anticancer activity of S. qinglanensis was tested against two different cell lines: HepG2 and CACO. The inhibition activities were 42.96 and 57.14%, respectively. Our findings suggest that the marine S. qinglanensis strain, which grows well on tailored medium, might be a source of bioactive substances for healthcare companies.

Failure Criteria of a 6-Inch Carbon Steel Pipe Elbow According to Deformation Angle Measurement Positions (변형각의 측정 위치에 따른 6인치 탄소강관엘보의 파괴 기준)

  • Yun, Da Woon;Jeon, Bub Gyu;Chang, Sung Jin;Park, Dong Uk;Kim, Sung Wan
    • Journal of the Earthquake Engineering Society of Korea
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    • v.26 no.1
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    • pp.13-22
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    • 2022
  • This study proposes a low-cycle fatigue life derived from measurement points on pipe elbows, which are components that are vulnerable to seismic load in the interface piping systems of nuclear power plants that use seismic isolation systems. In order to quantitatively define limit states regarding leakage, i.e., actual failure caused by low-cycle fatigue, in-plane cyclic loading tests were performed using a sine wave of constant amplitude. The test specimens consisted of SCH40 6-inch carbon steel pipe elbows and straight pipes, and an image processing method was used to measure the nonlinear behavior of the test specimens. The leakage lines caused by low-cycle fatigue and the low-cycle fatigue curves were compared and analyzed using the relationship between the relative deformation angles, which were measured based on each of the measurement points on the straight pipe, and the moment, which was measured at the center of the pipe elbow. Damage indices based on the combination of ductility and dissipation energy at each measurement point were used to quantitatively express the time at which leakage occurs due to through-wall cracking in the pipe elbow.

Determinants of Profitability of Regional Public Hospitals in Korea - Focusing on the COVID-19 Pandemic Period - (지역거점 공공병원의 수익성 결정요인 - COVID-19 유행기간을 중심으로 -)

  • Ji, Seokmin;Ok, Hyunmin
    • Korea Journal of Hospital Management
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    • v.27 no.3
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    • pp.26-38
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    • 2022
  • Purposes: We analyzed the profitability determinants of regional public hospitals during the entire period between 2010 and 2020 and the period before and after COVID-19. We intended to provide fundamental data for developing publicness evaluation index and task of establishing and expanding regional public hospitals. Methodology: The financial and non-financial information of the regional public hospitals were used as the main analysis data; The financial data was established by the Center for Public Healthcare Policy of National Medical Center, and the non-financial data by the Health Insurance Review and Assessment Service. T-test and regression analysis were used. Findings: The results can be summarized in two. First, the main determinants of profitability of the regional public hospitals were appeared to be the total asset turnover rate and the labor cost rate. Second, during the COVID-19 pandemic in the regional public hospitals, the number of sickbeds, the number of isolation rooms, the total asset turnover rate and the labor cost rate appeared to be the factor worsening the profitability. Practical Implication: The results of this study suggests that the management of the regional public hospitals is not aiming for the profit making, but it performs the functions as the community healthcare safety net such as controlling infectious diseases.

Decoding the intestinal microbiota repertoire of sow and weaned pigs using culturomic and metagenomic approaches

  • Mun, Daye;Kim, Hayoung;Shin, Minhye;Ryu, Sangdon;Song, Minho;Oh, Sangnam;Kim, Younghoon
    • Journal of Animal Science and Technology
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    • v.63 no.6
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    • pp.1423-1432
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    • 2021
  • To elucidate the role and mechanism of microbes, we combined culture-dependent and culture-independent approaches to investigate differences in gut bacterial composition between sows and weaned pigs. Under anaerobic conditions, several nonselective and selective media were used for isolation from fecal samples. All isolated bacteria were identified and classified through 16S rRNA sequencing, and the microbiota composition of the fecal samples was analyzed by metagenomics using next generation sequencing (NGS) technology. A total of 278 and 149 colonies were acquired from the sow and weaned pig fecal samples, respectively. Culturomics analysis revealed that diverse bacterial genus and species belonged to Firmicutes, Actinobacteria, Proteobacteria, and Bacteroidetes were isolated from sow and weaned pigs. When comparing culture-dependent and culture-independent analyses, 191 bacterial species and 2 archaeal bacterial species were detected through culture-independent analysis, and a total of 23 bacteria were isolated through a culture-dependent approach, of which 65% were not detected by metagenomics. In conclusion, culturomics and metagenomics should be properly combined to fully understand the intestinal microbiota, and livestock-derived microbial resources should be informed by culturomic approaches to understand and utilize the mechanism of host-microbe interactions.

Lysinabacillus fusiformis and Paenibacillus alvei Obtained from the Internal of NasutitermesTermites Revealed Their Ability as Antagonist of Plant Pathogenic Fungi

  • Fitriana, Yuyun;Tampubolon, Desi Apriani Teresa;Suharjo, Radix;Lestari, Puji;Swibawa, I Gede
    • The Plant Pathology Journal
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    • v.38 no.5
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    • pp.449-460
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    • 2022
  • This study was performed to reveal phenotypic characters and identity of symbiont bacteria of Nasutitermes as well as investigate their potential as antagonist of plant pathogenic fungi. Isolation of the symbiont bacteria was carried out from inside the heads and the bodies of soldier and worker termite which were collected from 3 locations of nests. Identification was performed using phenotypic test and sequence of 16S ribosomal DNA (16S rDNA). Antagonistic capability was investigated in the laboratory against 3 phytopathogenic fungi i.e., Phytophthora capsici, Ganoderma boninense, and Rigidoporus microporus. Totally, 39 bacterial isolates were obtained from inside the heads and the bodies of Nasutitermes. All the isolates showed capability to inhibit growth of P. capsici, however, 34 isolates showed capability to inhibit growth of G. boninense and 32 isolates showed capability to inhibit growth of R. microporus. Two bacterial strains (IK3.1P and 1B1.2P) which showed the highest percentage of inhibition were further identified based on their sequence of 16S rDNA. The result showed that 1K3.1P strain was placed in the group of type strain and reference strains of Lysinibacillus fusiformis meanwhile 1B1.2P strain was grouped within type strain and reference strains Paenibacillus alvei. The result of this study supply valuable information on the role of symbiont bacteria of Nasutitermes, which may support the development of the control method of the three above-mentioned phytopathogenic fungi.

Chemical Constituents of the Culture Broth of Panus rudis

  • Song, Ja-Gyeong;Ha, Lee Su;Ki, Dae-Won;Choi, Dae-Cheol;Lee, In-Kyoung;Yun, Bong-Sik
    • Mycobiology
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    • v.49 no.6
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    • pp.604-606
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    • 2021
  • In our ongoing search for new secondary metabolites from fungal strains, one novel compound (1) and nine known compounds (2-10) were isolated from the EtOAc-soluble layer of the culture broth of Panus rudis. The culture broth of P. rudis was extracted in acetone and fractionated by solvent partition; column chromatography using silica gel, Sephadex LH-20, and Sephadex G-10; MPLC; and HPLC. The structures of isolated compounds were elucidated by one- and two-dimensional NMR and LC-ESI-mass measurements. One new compound, panepoxydiol (1), and nine known compounds, (E)-3-(3-hydroxy-3-methylbut-1-en-1-yl)-7-oxabicyclo[4.1.0]hept-3-ene-2,5-diol (2), isopanepoxydone (3), neopanepoxydone (4), panepoxydone (5), panepophenanthrin (6), 4-hydroxy-2,2-dimethyl-6-methoxychromane (7), 6-hydroxy-2,2-dimethyl-3-chromen (8), 2,2-dimethyl-6-methoxychroman-4-one (9), 3,4-dihydroxy-2,2-dimethyl-6-methoxychromane (10), were isolated from the culture broth of P. rudis. This is the first report of isolation of a new compound panepoxydiol (1) and nine other chemical constituents (2-5, 7-10) from the culture broth of P. rudis.

A standardized method to study immune responses using porcine whole blood

  • Sameer-ul-Salam Mattoo;Ram Prasad Aganja;Seung-Chai Kim;Chang-Gi Jeong;Salik Nazki;Amina Khatun;Won-Il Kim;Sang-Myeong Lee
    • Journal of Veterinary Science
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    • v.24 no.1
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    • pp.11.1-11.14
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    • 2023
  • Background: Peripheral blood mononuclear cells (PBMCs) are commonly used to assess in vitro immune responses. However, PBMC isolation is a time-consuming procedure, introduces technical variability, and requires a relatively large volume of blood. By contrast, whole blood assay (WBA) is faster, cheaper, maintains more physiological conditions, and requires less sample volume, laboratory training, and equipment. Objectives: Herein, this study aimed to develop a porcine WBA for in vitro evaluation of immune responses. Methods: Heparinized whole blood (WB) was diluted (non-diluted, 1/2, 1/8, and 1/16) in RPMI-1640 media, followed by phorbol myristate acetate and ionomycin. After 24 h, cells were stained for interferon (IFN)-γ secreting T-cells followed by flow cytometry, and the supernatant was analyzed for tumor necrosis factor (TNF)-α. In addition, diluted WB was stimulated by lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C), reference strain KCTC3557 (RS), field isolate (FI), of heat-killed (HK) Streptococcus suis, and porcine reproductive and respiratory syndrome virus (PRRSV). Results: The frequency of IFN-γ+CD3+ T-cells and concentration of TNF-α in the supernatant of WB increased with increasing dilution factor and were optimal at 1/8. WB TNF-α and interleukin (IL)-10 cytokine levels increased significantly following stimulation with LPS or poly I:C. Further, FI and RS induced IL-10 production in WB. Additionally, PRRSV strains increased the frequency of IFN-γ+ CD4-CD8+ cells, and IFN-γ was non-significantly induced in the supernatant of re-stimulated samples. Conclusions: We propose that the WBA is a rapid, reliable, and simple method to evaluate immune responses and WB should be diluted to trigger immune cells.

Secondary Metabolites from Anthonotha cladantha (Harms) J.Léonard

  • Laurent Voufack Lefack Bongmo;Achille Nouga Bissoue;Samuel Magloire Bissim;Georges Bellier Tabekoueng;Willifred Dongmo Tekapi Tsopgni;Mehreen Lateef;Felicien Mushagalusa Kasali;Muhammad Shaiq Ali;Alain Francois Kamdem Waffo;Jean Duplex Wansi
    • Natural Product Sciences
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    • v.29 no.1
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    • pp.50-58
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    • 2023
  • The phytochemical investigation of the crude methanolic extracts roots and stem bark of Anthonotha cladantha (Harms) J.Léonard led to the isolation and identification of twelve secondary metabolites: 2,3-dihydroxypropyl hexacosanoate (1), hederagenine (2), cycloeucalenol (3), 2α-hydroxylupeol (4), betulinic acid (5), lupeol (6), heptacosan-2-one (7), triacontanoic acid (8), stigmast-4-en-3-one (9), β-sitosterol (10), stigmasterol (11), and stigmasterol-3-O-β-D-glucopyranoside (12). Their structures were elucidated with the help of their spectroscopic and physical data and by comparison with those reported in the literature. To the best of our knowledge, from all those compounds, 2,3-dihydroxypropyl hexacosanoate (1), hederagenine (2), cycloeucalenol (3), 2α-hydroxylupeol (4), and betulinic acid (5) are being reported for the first time from this genus. In addition, the acetylation of compound 1 afforded a new derivative 3-(hexacosanoyloxy)propane-1,2-diyl diacetate (1a). Compound 1 possessed a moderate α-glucosidase inhibitory activity with an IC50 value of 39.2 ± 0.22 μM; it neither showed antioxidant activity nor inhibition against the enzyme urease. Compound 1a exhibited weak antioxidant activity in the DPPH assay with an IC50 value of 80.3 ± 0.83 μM but was inactive against α-glucosidase and urease. Furthermore, both compounds 1 and 1a were inactive against seven pathogenic bacterial strains.

The effects of gingival blood flow on pulpal blood flow detection using ultrasound Doppler flowmetry: animal study

  • Dohyun Kim ;Hyoung-Seok Ko;Soo-Yeon Park ;Seung-Yeon Ryu ;Sung-ho Park
    • Restorative Dentistry and Endodontics
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    • v.48 no.1
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    • pp.9.1-9.11
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    • 2023
  • Objectives: This study evaluated the effect of adjacent gingival blood flow on detection of pulpal blood flow (PBF) using ultrasound Doppler flowmetry (UDF) through animal study. Materials and Methods: The study included 36 right and left maxillary the third incisors and canines in 9 experimental dogs. The study included 2 main steps: In the first step, the pulse sound level (PSL) was recorded on the cervical part of each tooth without flap elevation (Group 1), with flap elevation (Group 2), and after it was repositioned in place (Group 3). In the second step, the PSL was recorded on the cervical part of each tooth (Group 4), after pulpotomy (Group 5), after partial pulp extirpation (Group 6), after complete extirpation (Group 7), and after canal filling (Group 8). In Groups 5-8, the study was performed with and without flap elevation in the left and right teeth, respectively. The PSL was graded as follows: 0, inaudible; 1, heard faintly; and 2, heard well. The difference between each group was analyzed using Friedman's test with Wilcoxon signed-rank tests (α = 0.05). Results: In step 1, the PSL results were Group 1 > 2 and 3. In step 2, there was no significant difference between the groups when the flap was not elevated, while PSL results were Group 4 > 5 ≥ 6 and 7 ≥ 8 when the flap was elevated. Conclusions: PBF is affected by gingival blood flow when measured with UDF. UDF measurements require isolation of gingiva from the tooth.

The potential of non-movement behavior observation method for detection of sick broiler chickens

  • Hyunsoo Kim;Woo-Do Lee;Hyung-Kwan Jang;Min Kang;Hwan-Ku Kang
    • Journal of Animal Science and Technology
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    • v.65 no.2
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    • pp.441-458
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    • 2023
  • The poultry industry, which produces excellent sources of protein, suffers enormous economic damage from diseases. To solve this problem, research is being conducted on the early detection of infection according to the behavioral characteristics of poultry. The purpose of this study was to evaluate the potential of a non-movement behavior observation method to detect sick chickens. Forty 1-day-old Ross 308 males were used in the experiments, and an isolator equipped with an Internet Protocol (IP) camera was fabricated for observation. The chickens were inoculated with Salmonella enterica serovar Gallinarum A18-GCVP-014, the causative agent of fowl typhoid (FT), at 14 days of age, which is a vulnerable period for FT infection. The chickens were continuously observed with an IP camera for 2 weeks after inoculation, chickens that did not move for more than 30 minutes were detected and marked according to the algorithm. FT infection was confirmed based on clinical symptoms, analysis of cardiac, spleen and liver lesion scores, pathogen re-isolation, and serological analysis. As a result, clinical symptoms were first observed four days after inoculation, and dead chickens were observed on day six. Eleven days after inoculation, the number of clinical symptoms gradually decreased, indicating a state of recovery. For lesion scores, dead chickens scored 3.57 and live chickens scored 2.38. Pathogens were re-isolated in 37 out of 40 chickens, and hemagglutination test was positive in seven out of 26 chickens. The IP camera applied with the algorithm detected about 83% of the chickens that died in advance through non-movement behavior observation. Therefore, observation of non-movement behavior is one of the ways to detect infected chickens in advance, and it appears to have potential for the development of remote broiler management system.