• Title/Summary/Keyword: NP0

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In silico annotation of a hypothetical protein from Listeria monocytogenes EGD-e unfolds a toxin protein of the type II secretion system

  • Maisha Tasneem;Shipan Das Gupta;Monira Binte Momin;Kazi Modasser Hossain;Tasnim Binta Osman;Fazley Rabbi
    • Genomics & Informatics
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    • v.21 no.1
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    • pp.7.1-7.11
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    • 2023
  • The gram-positive bacterium Listeria monocytogenes is an important foodborne intracellular pathogen that is widespread in the environment. The functions of hypothetical proteins (HP) from various pathogenic bacteria have been successfully annotated using a variety of bioinformatics strategies. In this study, a HP Imo0888 (NP_464414.1) from the Listeria monocytogenes EGD-e strain was annotated using several bioinformatics tools. Various techniques, including CELLO, PSORTb, and SOSUIGramN, identified the candidate protein as cytoplasmic. Domain and motif analysis revealed that the target protein is a PemK/MazF-like toxin protein of the type II toxin-antitoxin system (TAS) which was consistent with BLASTp analysis. Through secondary structure analysis, we found the random coil to be the most frequent. The Alpha Fold 2 Protein Structure Prediction Database was used to determine the three-dimensional (3D) structure of the HP using the template structure of a type II TAS PemK/MazF family toxin protein (DB ID_AFDB: A0A4B9HQB9) with 99.1% sequence identity. Various quality evaluation tools, such as PROCHECK, ERRAT, Verify 3D, and QMEAN were used to validate the 3D structure. Following the YASARA energy minimization method, the target protein's 3D structure became more stable. The active site of the developed 3D structure was determined by the CASTp server. Most pathogens that harbor TAS create a crucial risk to human health. Our aim to annotate the HP Imo088 found in Listeria could offer a chance to understand bacterial pathogenicity and identify a number of potential targets for drug development.

Comparative evaluation of gold nanoparticles and Alum as immune enhancers against rabies vaccine and related immune reactivity, physiological, and histopathological alterations: in vivo study

  • Rehab Essam El-Din El-Hennamy;Sahar Mohamed Mahmoud;Nabil Ahmed El-Yamany;Hanaa Hassan Hassanein;Mohamed Elsayed Amer;Aly Fahmy Mohamed
    • Clinical and Experimental Vaccine Research
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    • v.12 no.1
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    • pp.32-46
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    • 2023
  • Purpose: The present study aimed to compare the immune-enhancing potential of gold nanoparticles (AuNPs) to Alum against rabies vaccine and the related immunological, physiological, and histopathological effects. Materials and Methods: Alum and AuNPs sole and in combination with rabies vaccine were used at 0.35 mg/mL and 40 nM/mL, respectively. Rats used were categorized into six groups (20/each): control rats, rabies vaccine, aluminum phosphate gel, rabies vaccine adsorbed to Alum, AuNPs, and rabies vaccine adjuvant AuNPs. Results: Liver and kidney functions were in the normal range after AuNPs and Alum adjuvanted vaccine compared to control. Interleukin-6 and interferon-γ levels were significantly increased in groups immunized with Alum and AuNPs adjuvanted vaccine, the peak level was in the case of AuNP adjuvanted vaccine on the 14th day. Ninety days post-vaccination, total immunoglobulin G (IgG) against adjuvanted rabies vaccine showed a significantly elevated antirabies IgG with AuNPs and Alum adsorbed vaccine compared with unadjuvanted one. The total antioxidant capacity, malondialdehyde (MDA) levels, superoxide dismutase, and glutathione peroxidase activities were significantly increased post-adjuvanted AuNPs adjuvanted vaccine vaccination than in Alum adsorbed vaccine, while MDA was significantly decreased. The histopathological examination revealed detectable alterations post-AuNPs and Alum adjuvanted vaccine immunization compared with liver and kidney profiles post-administration of unadjuvanted and non-immunized groups, meanwhile, splenic tissue revealed hyperplasia of lymphoid follicles indicating increased immune reactivity. Conclusion: The AuNPs are promising enhancers of the immune response as Alum, and the undesirable effects of AuNPs could be managed by using suitable sizes, shapes, and concentrations.

Analysis of Spatial Changes in the Forest Landscape of the Upper Reaches of Guem River Dam Basin according to Land Cover Change (토지피복변화에 따른 금강 상류 댐 유역 산림 경관의 구조적 변화 분석)

  • Kyeong-Tae Kim;Hyun-Jung Lee;Whee-Moon Kim;Won-Kyong Song
    • Korean Journal of Environment and Ecology
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    • v.37 no.4
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    • pp.289-301
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    • 2023
  • Forests within watersheds are essential in maintaining ecosystems and are the central infrastructure for constructing an ecological network system. However, due to indiscriminate development projects carried out over past decades, forest fragmentation and land use changes have accelerated, and their original functions have been lost. Since a forest's structural pattern directly impacts ecological processes and functions in understanding forest ecosystems, identifying and analyzing change patterns is essential. Therefore, this study analyzed structural changes in the forest landscape according to the time-series land cover changes using the FRAGSTATS model for the dam watershed of the Geum River upstream. Land cover changes in the dam watershed of the Geum River upstream through land cover change detection showed an increase of 33.12 square kilometers (0.62%) of forests and 67.26 square kilometers (1.26%) of urbanized dry areas and a decrease of 148.25 square kilometers (2.79%) in agricultural areas from the 1980s to the 2010s. The results of no-sampling forest landscape analysis within the watershed indicated landscape percentage (PLAND), area-weighted proximity index (CONTIG_AM), average central area (CORE_MN), and adjacency index (PLADJ) increased, and the number of patches (NP), landscape shape index (LSI), and cohesion index (COHESION) decreased. Identification of structural change patterns through a moving window analysis showed the forest landscape in Sangju City, Gyeongsangbuk Province, Boeun County in Chungcheongbuk Province, and Jinan Province in Jeollabuk Province was relatively well preserved, but fragmentation was ongoing at the border between Okcheon County in Chungcheongbuk Province, Yeongdong and Geumsan Counties in Chungcheongnam Province, and the forest landscape in areas adjacent to Muju and Jangsu Counties in Jeollabuk Province. The results indicate that it is necessary to establish afforestation projects for fragmented areas when preparing a future regional forest management strategy. This study derived areas where fragmentation of forest landscapes is expected and the results may be used as basic data for assessing the health of watershed forests and establishing management plans.

Epidemiological Observation on Recent Outbreaks of Canine Distemper in Korea (최근 국내발병 개 디스템퍼에 대한 역학적 조사)

  • Doo, Kim;Ji-young, Park;So-jeo, Ahn;Seok-young, Jeoung;Son-il, Pak
    • Journal of Veterinary Clinics
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    • v.21 no.3
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    • pp.229-235
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    • 2004
  • To characterize the recent outbreaks of canine distemper (CD) in Korea, we carried out epidemiological investigations by clinical observations, serum neutralizing (SN) antibody titer determination and RT-PCR on the 315 dogs which were clinically suspected as canine distemper virus (CDV) infection. One hundred and sixty two of 315 dogs were infected with CDV. Breed or gender did not seem to have effects on the prevalence of CD. The major part of dogs were in young age from 6 weeks to 18 weeks of age, and were not vaccinated or incompletely vaccinated. Clinical signs of dogs with CD were multi systemic and extremely variable. Dogs died from CD had significantly more ocular signs and neurologic signs than those of dogs survived (p<0.05). The SN titers against CDV of 157 (96.9%) dogs were under 1:16, which is less than protective level. One possible explanation for recent outbreaks of CD in Korea might be low antibody titers against CDV because of vaccination failure. Therefore, to reduce the impact of virulent infection in the dog population, dogs should be vaccinated adequately and prophylactic measures should include isolation of young dogs from the dog population until vaccination can be expected to provide protection.

Effects of Size, Impurities, and Citrate Capping on the Toxicity of Manufactured Silver Nano-particles to Larval Zebrafish (Danio rerio)

  • Kim, Jungkon;Park, Yena;Lee, Sangwoo;Seo, Jihyun;Kwon, Dongwook;Park, Jaehong;Yoon, Tae-Hyun;Choi, Kyungho
    • Journal of Environmental Health Sciences
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    • v.39 no.4
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    • pp.369-375
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    • 2013
  • Objectives: This study was conducted to identify factors determining the toxicity of manufactured silver nano-particles (AgNPs) on aquatic organisms. Methods: For this purpose, we prepared several AgNPs with varied characteristics, including hydrodynamic size (nano-$^{ABC}Ag^{Cit}\;vs$-sized-$^{ABC}Ag^{Cit}$), impurities ($^{ABC}Ag$ stock vs $^{ABC}Ag$), and citrate capping ($^{ABC}Ag^{Cit}$), using a commercially available manufactured AgNP ($^{ABC}Ag$ stock). Acute tests were conducted using larval zebrafish (Danio rerioI). In addition, in order to determine the ecotoxicological potentials of various capping agents, toxicity tests were conducted with microbes, waterfleas, and fish for eight different capping agents that are used for NPs. Results: The toxicity of AgNPs in terms of 96 h fish $LC_{50}$ increased in the following order: $^{ABC}Ag$ stock < $^{ABC}Ag=^{ABC}Ag^{Cit}=nano-^{ABC}Ag^{Cit}$ < ${\mu}$-sized-$^{ABC}Ag^{Cit}$ < $AgNO_3$. After removing impurities by dialysis, 96 h $LC_{50}$ value decreased significantly from $126.6{\mu}g/L$ (95% confidence intervals [CI]: 107.0-146.2) ($^{ABC}Ag$ stock) to $78.6{\mu}g/L$ (CI: 72.7-84.8) ($^{ABC}Ag$). For ${\mu}$-sized-$^{ABC}Ag^{Cit}$ (ranging between 3.9 and 40.6 nm) and $^{ABC}Ag^{Cit}$ (40.6 nm and $9.1{\mu}m$), the 96 h $LC_{50}$ of the former ($43.9{\mu}g/L$, CI: 36.0-51.7) was approximately two-fold lower than that of the latter ($87.0{\mu}g/L$, CI: 73.5-100.3). Conclusions: In this study, we found that for acute lethality, the contribution of impurities and particle size was significant, but that of citrate was negligible.

Isolation and Characterization of Acinetobacter sp. BD5 Producing Lipolytic Enzyme (Lipolytic 효소를 생산하는 Acinetobacter sp. BD5 균주의 분리 및 특성)

  • Park, In-Hye;Kim, Sun-Hee;Lee, Sang-Cheol;Ahn, Soon-Cheol;Kim, Cheol-Min;Choi, Yong-Lark
    • Journal of Life Science
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    • v.16 no.4
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    • pp.555-560
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    • 2006
  • A bacterium producing novel lipolytic enzyme was isolated from house sewage and identified as Acinetobacter sp. BD5 based on physiological characterization and 16S rDNA sequencing. The lipolytic activity of Acinetobacter sp. BD5 was tested using an EL agar medium and CE agar medium supplemented with 1% tributyrin and olive oil, respectively. The formation of a clear zone around the colony was detected by agar medium supplemented with 1% tributyrin and olive oil, respectively and Acinetobacter sp. BD5 formed powder-like zone around the colony on LB agar medium containing Tween 20. The quantitative lipolytic activity was determined by using p-NP butyrate as substrate. Acinetobacter sp. BD5 secreted the lipolytic enzyme during exponential growth phase, reaching a maximum amount after 6 hours of incubation. The lipolytic enzyme was found to be optimally active at $60^{\circ}C$ and retained more than 70% at $70-80^{\circ}C$. It displayed a high degree of activity in a pH of 7.0 to 10.6, with an optimal pH of 9.0.

Investigation of Nonylphenols Contamination in Solvents and Solid-phase Extraction Cartridge, and its Removal Protocols (정밀분석용 용매 및 SPE의 Nonylphenols 오염평가 및 제거)

  • Park, Jong-Min;Choi, Geun-Hyonng;Kim, Jung-Im;Hong, Su-Myeong;Kwon, Oh-Kyung;Im, Geon-Jae;Kim, Jin-Hyo
    • The Korean Journal of Pesticide Science
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    • v.15 no.1
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    • pp.22-27
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    • 2011
  • Nonylphenols are toxic compounds classified as endocrine disruptors. We investigated the nonylphenols clean-up procedures for the contamination control in the quantitative analysis. In this research we analyzed the residual nonylphenols in the solvent and the SPE cartridges. First, at the analysis of HPLC grade solvents (n-hexane, diethyl ether, ethyl acetate and its mixture), diethyl ether was confirmed the residue as 0.963 ${\mu}g/mL$, and we eliminated the contaminant through the distillation with $CaH_2$, Second, at the analysis of SPE cartridges (silica gel and Florisil), all products were showed the residue at 0.046~13.0 ${\mu}g/mL$, but unfortunately the residue in the cartridge were not easily removed with referenced methods in all tested SPE cartridges except in silica gel SPE cartridge with glass ware.

Optimization of Protoplast Isolation and Ribonucleoprotein/Nanoparticle Complex Formation in Lentinula edodes (표고버섯의 원형질체 분리 최적화와 RNPs/나노파티클 복합체 형성)

  • Kim, Minseek;Ryu, Hojin;Oh, Min Ji;Im, Ji-Hoon;Lee, Jong-Won;Oh, Youn-Lee
    • Journal of Mushroom
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    • v.20 no.3
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    • pp.178-182
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    • 2022
  • Despite the long history of mushroom use, studies examining the genetic function of mushrooms and the development of new varieties via bio-molecular methods are significantly lacking compared to those examining other organisms. However, owing to recent developments, attempts have been made to use a novel gene-editing technique involving CRISPR/Cas9 technology and genetic scissors in mushroom studies. In particular, research is actively being conducted to utilize ribonucleoprotein particles (RNPs) that can be genetically edited with high efficiency without foreign gene insertion for ease of selection. However, RNPs are too large for Cas9 protein to pass through the cell membrane of the protoplasmic reticulum. Furthermore, guide RNA is unstable and can be easily decomposed, which remarkably affects gene editing efficiency. In this study, nanoparticles were used to mitigate the shortcomings of RNP-based gene editing techniques and to obtain transformants stably. We used Lentinula edodes (shiitake mushroom) Sanjo705-13 monokaryon strain, which has been successfully used in previous genome editing experiments. To identify a suitable osmotic buffer for the isolation of protoplast, 0.6 M and 1.2 M sucrose, mannitol, sorbitol, and KCl were treated, respectively. In addition, with various nanoparticle-forming materials, experiments were conducted to confirm genome editing efficiency via the formation of nanoparticles with calcium phosphate (CaP), which can be bound to Cas9 protein without any additional amino acid modification. RNPs/NP complex was successfully formed and protected nuclease activity with nucleotide sequence specificity.

A Comparative Study of Physiological Activity and Ingredient Analysis of Glycyrrhiza uralensis Fischer Stems and Leaves Cultivated with Different Wavelength of LED Lights (LED광원에 따른 감초 지상부의 생리활성 및 성분 평가)

  • Bang, Keuk Soo;Chang, Young Nam;Jin, Jong Sik;Park, Sang A;Lim, Jae Soo;Park, Jeong Sub;Kim, Jong Sung;Lee, Jeong Ho
    • Korean Journal of Plant Resources
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    • v.28 no.1
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    • pp.126-134
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    • 2015
  • Glycyrrhiza uralensis Fischer is one of the most commonly used herbs. Recently, the stem and leave of the plant have been interested in physiological activities because the aerial parts have been thrown away. Finding out cultivation method of Glycyrrhiza uralensis Fischer to improve chemical ingredients and biological activities has been tried these days. In this study, different wavelengths of light emitting diode (LED) were used for a cultivation of Glycyrrhiza uralensis Fischer. Antioxidant activities and inhibitory effect on mutagenecity of samples were evaluated. The stem and leave cultivated under blue light (BL-0) showed the strongest antioxidant activities of $3.02{\pm}0.13{\mu}g/ml$ ($EC_{50}$) and $2.18{\pm}0.18{\mu}g/ml$ ($EC_{50}$) in DPPH and ABTS radical scavenging test, respectively. Total phenolic content of BL-0 was $2.93{\pm}0.11g/100g$, the highest value between cultivation conditions. However, antioxidant activities of the stem and leave cultivated under red light were the weakest between samples. All of the stem and leave used in this study showed inhibitory effect on mutagenecity of 1-nitropyrene. BL-0 showed stronger inhibitory effects on mutagenicity of Trp-P-1, Trp-P-2, and AFB1 than samples cultivated under other conditions. Only on mutagenecity of 2-aminoanthracene, the stem and leave cultivated at 1 m apart from red light (RL-1) showed the strongest inhibitory effect. These results indicate that blue LED might be the most effective condition for improvement of physiological activities for the aerial parts of Glycyrrhiza uralensis Fischer in cultivation. The components were identified with GC/MS. Cytidine was detected only in RL-1 at 25 min of retention time and 2-bromotrimethylene glycol was detected only in BL-0 at 37 min.

Fabrication of Gd2O2S:Tb fine scintillator film and evaluation of image quality for resolution improvement of X-ray imaging based on CMOS (CMOS 기반 X선 영상의 해상력 향상을 위한 Gd2O2S:Tb 미세형광체 필름 제작 및 영상 질 평가)

  • Kang, Sang-Sik;Choi, Young-Zoon;Jung, Bong-Jae;No, Si-Cheul;Cho, Chan-Hoon;Yoon, In-Chan;Park, Ji-Koon
    • Journal of the Korean Society of Radiology
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    • v.5 no.5
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    • pp.283-287
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    • 2011
  • In this study, fine $Gd_2O_2S$:Tb powder was synthesized by using a low temperature solution-combustion method for a high-resolution digital x-ray imaging detector. From the fabricated phosphor power, the fine scintillator films was fabricated by particle sedimentation method and was investigated the luminescent property. From the experimental results of relative light output as a function of terbium concentration, the highest luminescent efficiency has at 5 wt% Tb concentration, and luminescent intensity decreased rapidly according to quenching effect about higher Tb concentration. Also, the relative light output of $270{\mu}m$-$Gd_2O_2S$:Tb film has 2945 pC/$cm^2$/mR. And light intensity was saturated at higher film thickness. Finally, to evaluate an image acquisition performance of fabricated phosphor, images were obtained by using commercial CMOS sensor and measured the MTF, NPS, and DQE. DQE(0 lp/mm) of fine phosphor film has 37%. But, DQE improvement of fine phosphor film is possible by resolving problem of film fabrication process and has a significant potential in the application of digital radiation imaging system later.