• Title/Summary/Keyword: Single-Cell Analysis

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Fabrication and Characteristics of Porous Silicon (다공성 실리콘의 제조 및 특성에 관한 연구)

  • 이철환;조원일;백지흠;박성용;안춘호;유종훈;조병원;윤경석
    • Journal of Surface Science and Engineering
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    • v.28 no.3
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    • pp.182-191
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    • 1995
  • A highly porous silicon layer was fabricated by anodizing single crystalline silicon in a dilute solution of hydrofluoric acid. The color of the porous silicon changed from red and blue to yellow gold during the anodizing process. The current-voltage (I-V) curve of the anodizing process showed a typical Schottky diode rectification form. The cell voltage decreased with the increase of HF concentration in the solution at high current range. However, the voltage was independent on HF concentration in the solution at low current range. The pore size was dependant on anodizing condition (HF concentration, current and anodizing time). The pore size and wall width of porous silicon layer were 4~6 and 1~3 nm, respectively. Surface of the porous silicon was covered with silicon compound ($SiH_x$etc.) according to IR spectrum analysis. The peak wavelength and width of photoluminescence (PL) spectrum of porous silicon were 650~850 nm (1.5~1.9 eV) and 250 nm, respectively. The photoluminescence intensity and peak wavelength, and porosity of porous silicon increased with increasing anodizing current and decreased with increasing HF concentration in the anodizing solution.

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High-Efficiency Generation of Monoclonal Antibody for Vitreoscilla Hemoglobin Protein

  • Kim, Eun-Mi;Kim, Myung-Hee;Kim, Min-Gon;Kim, Sang-Woo;Ro, Hyeon-Su
    • Journal of Microbiology and Biotechnology
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    • v.22 no.2
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    • pp.226-229
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    • 2012
  • Bacterial hemoglobin from Vitreoscilla (VHb) is recognized as a good fusion protein for the soluble expression of foreign protein. In this study, we generated a monoclonal antibody (MAb) against VHb for its detection. For the rapid screening of MAb, a protein chip technology based on the Alexa-488 (A488) dye labeling method was introduced. In order to fabricate the chip, the VHb protein was chemically coupled to the chip surface and then the culture supernatants of 84 hybridoma cell lines were spotted onto the VHb chip. The bound MAbs were measured by A488-modified anti-mouse IgG. A single spot (MAb A10) exhibited significantly high signal intensity. The immunoblot analysis evidenced that the MAb A10 can detect VHb-fused proteins with high specificity.

The Analysis of Priority Output Queuing Model by Short Bus Contention Method (Short Bus contention 방식의 Priority Output Queuing Model의 분석)

  • Jeong, Yong-Ju
    • The Transactions of the Korea Information Processing Society
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    • v.6 no.2
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    • pp.459-466
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    • 1999
  • I broadband ISDN every packet will show different result if it would be processed according to its usage by the server. That is, normal data won't show big differences if they would be processed at normal speed. But it will improve the quality of service to process some kinds of data - for example real time video or voice type data or some data for a bid to by something through the internet - more fast than the normal type data. solution for this problem was suggested - priority packets. But the analyses of them are under way. Son in this paper a switching system for an output queuing model in a single server was assumed and some packets were given priorities and analysed. And correlation, simulating real life situation, was given too. These packets were analysed through three cases, first packets having no correlation, second packets having only correlation and finally packets having priority three cases, first packets having no correlation, second packets having only correlation and finally packets having priority and correlation. The result showed that correlation doesn't affect the mean delay time and the high priority packets have improved mean delay time regardless of the arrival rate. Those packets were assumed to be fixed-sized like ATM fixed-sized cell and the contention strategy was assumed to be short bus contention method for the output queue, and the mean delay length and the maximum 버퍼 length not to lose any packets were analysed.

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First Report of Gray Mold Disease on Endangered Species Cypripedium japonicum

  • Jeon, Chang-Wook;Kim, Da-Ran;Gang, Geun Hye;Kim, Byung-Bu;Kim, Nam Ho;Nam, Seong-Yeol;Kwak, Youn-Sig
    • Mycobiology
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    • v.48 no.5
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    • pp.423-426
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    • 2020
  • Cypripedium japonicum is known to be the indigenous plant to Korea, Japan, and China. However, C. japonicum represents the most critically endangered plant species in South Korea. The plant is esthetically pleasing due to its flower, which is larger than any other orchidaceous species. Disease symptoms relating to gray mold were observed on C. japonicum in May 2019. The suspected pathogen was successfully isolated from the symptomatic leaf tissue and conducted a pure culture of the fungi. The conidia formed consisted of a colorless or light brown single cell, which was either egg or oval-shaped with a size of 7.1 to 13.4 × 5.2 to 8.6 ㎛. Molecular phylogenetic relationship analysis was also confirmed that the pathogen concerned belonging to the family of Botrytis cinerea. Therefore, the findings confirmed that the pathogen isolated from C. japonicum was consistent with the unique properties of B. cinerea.

Effective Blocking of Microbial Transcriptional Initiation by dCas9-NG-Mediated CRISPR Interference

  • Kim, Bumjoon;Kim, Hyun Ju;Lee, Sang Jun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.12
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    • pp.1919-1926
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    • 2020
  • CRISPR interference (CRISPRi) has been developed as a transcriptional control tool by inactivating the DNA cleavage ability of Cas9 nucleases to produce dCas9 (deactivated Cas9), and leaving dCas9 the ability to specifically bind to the target DNA sequence. CRISPR/Cas9 technology has limitations in designing target-specific single-guide RNA (sgRNA) due to the dependence of protospacer adjacent motif (PAM) (5'-NGG) for binding target DNAs. Reportedly, Cas9-NG recognizing 5'-NG as the PAM sequence has been constructed by removing the dependence on the last base G of PAM through protein engineering of Cas9. In this study, a dCas9-NG protein was engineered by introducing two active site mutations in Cas9-NG, and its ability to regulate transcription was evaluated in the gal promoter in E. coli. Analysis of cell growth rate, D-galactose consumption rate, and gal transcripts confirmed that dCas9-NG can completely repress the promoter by recognizing DNA targets with PAM of 5'-NGG, NGA, NGC, NGT, and NAG. Our study showed possible PAM sequences for dCas9-NG and provided information on target-specific sgRNA design for regulation of both gene expression and cellular metabolism.

A Review of Rat Models of Avascular Necrosis of the Femoral Head Treated with Natural Extracts

  • Go-Woon, Kim;Hyoung-Yong, Park;Yeon-Cheol, Park
    • Journal of Acupuncture Research
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    • v.39 no.4
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    • pp.239-248
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    • 2022
  • To determine the effect of Korean medicine treatment of avascular necrosis of the femoral head (ANFH) this study reviewed both single ingredients and bioactive compounds in the treatment of ANFH in a rat model. Literature was retrieved from PubMed and Google Scholar using the keywords "femur head necrosis," "natural extract," and "rat." The data from studies analyzed included: rats' characteristics, development methods of ANFH, natural extracts administration, observation methods, and outcome indicators. Two independent researchers screened all articles retrieved and 26 studies were chosen. The most used rat species was the Sprague Dawley rat (76.9%). To induce ANFH, steroid injections (46.2%), and oral gavage (53.8%) were typically used. Studies focused mainly on factors affecting bone formation (65.3%), and apoptosis (53.8%). Research on ANFH focused on using traditional natural substances mentioned in classical literature to confirm its effectiveness against anti-inflammation, osteogenesis, and cell death. ANFH has a diverse etiology, therefore research models such as genetic analysis of human-derived samples from ANFH patients may shed more light on the condition. Moreover, research into herbal medicines and pharmacoacupuncture treatment of ANFH should precede.

Two Litonotid Ciliates (Ciliophora: Litostomatea: Pleurostomatida) Unknown from Korea

  • Lee, Jung-Mi;Yoon, Jae-Sool;Shin, Mann-Kyoon
    • Animal Systematics, Evolution and Diversity
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    • v.22 no.2
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    • pp.223-229
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    • 2006
  • Two litonotid ciliates collected from the freshwater habitats in Korea were identified as Loxophyllum meleagris ($M\ddot{u}ller$, 1773) and Siroloxophyllum utriculariae (Penard, 1922). The description was based on the observation of living and protargol impregnated specimens, and biometric analysis. Their diagnostic characteristics are as follows. L. meleagris; $163-480\times80-100{\mu}m$ in vivo, body shape lancet or knife-like; dorsal margin with 10-19 extrusome warts; 8-35 macronuclei nodules, like a string of bead; 16-21 somatic kineties on right side (including perioral kinety 2, 3) and 6-11 on left side (including perioral kinety 1); 1 contractile vacuole located at posterior part at diastole stage, extending along the dorsal margin toward anterior end with a single long narrow canal. S. utriculariae; $110-170\times78-150{\mu}m$ in vivo, body shape lancet like; dorsal margin without extrusome warts; 2 macronuclei, spherical; 12-19 somatic kineties on right side, 3-7 on left side (including perioral kinety 1); 2-3 contractile vacuoles, first one located anterior ventrally, second one located posterior dorsally and last one located near posterior end of cell.

Functional annotation of uncharacterized proteins from Fusobacterium nucleatum: identification of virulence factors

  • Kanchan Rauthan;Saranya Joshi;Lokesh Kumar;Divya Goel;Sudhir Kumar
    • Genomics & Informatics
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    • v.21 no.2
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    • pp.21.1-21.14
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    • 2023
  • Fusobacterium nucleatum is a gram-negative bacteria associated with diverse infections like appendicitis and colorectal cancer. It mainly attacks the epithelial cells in the oral cavity and throat of the infected individual. It has a single circular genome of 2.7 Mb. Many proteins in F. nucleatum genome are listed as "Uncharacterized." Annotation of these proteins is crucial for obtaining new facts about the pathogen and deciphering the gene regulation, functions, and pathways along with discovery of novel target proteins. In the light of new genomic information, an armoury of bioinformatic tools were used for predicting the physicochemical parameters, domain and motif search, pattern search, and localization of the uncharacterized proteins. The programs such as receiver operating characteristics determine the efficacy of the databases that have been employed for prediction of different parameters at 83.6%. Functions were successfully assigned to 46 uncharacterized proteins which included enzymes, transporter proteins, membrane proteins, binding proteins, etc. Apart from the function prediction, the proteins were also subjected to string analysis to reveal the interacting partners. The annotated proteins were also put through homology-based structure prediction and modeling using Swiss PDB and Phyre2 servers. Two probable virulent factors were also identified which could be investigated further for potential drug-related studies. The assigning of functions to uncharacterized proteins has shown that some of these proteins are important for cell survival inside the host and can act as effective drug targets.

Lactobacillus Persisters Formation and Resuscitation

  • Hyein Kim;Sejong Oh;Sooyeon Song
    • Journal of Microbiology and Biotechnology
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    • v.34 no.4
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    • pp.854-862
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    • 2024
  • Lactobacillus is a commonly used probiotic, and many researchers have focused on its stress response to improve its functionality and survival. However, studies on persister cells, dormant cells that aid bacteria in surviving general stress, have focused on pathogenic bacteria that cause infection, not Lactobacillus. Thus, understanding Lactobacillus persister cells will provide essential clues for understanding how Lactobacillus survives and maintains its function under various environmental conditions. We treated Lactobacillus strains with various antibiotics to determine the conditions required for persister formation using kill curves and transmission electron microscopy. In addition, we observed the resuscitation patterns of persister cells using single-cell analysis. Our results show that Lactobacillus creates a small population of persister cells (0.0001-1% of the bacterial population) in response to beta-lactam antibiotics such as ampicillin and amoxicillin. Moreover, only around 0.5-1% of persister cells are heterogeneously resuscitated by adding fresh media; the characteristics are typical of persister cells. This study provides a method for forming and verifying the persistence of Lactobacillus and demonstrates that antibiotic-induced Lactobacillus persister cells show characteristics of dormancy, sensitivity of antibiotics, same as exponential cells, multi-drug tolerance, and resuscitation, which are characteristics of general persister cells. This study suggests that the mechanisms of formation and resuscitation may vary depending on the characteristics, such as the membrane structure of the bacterial species.

Studies on the Petroleum hydrocarbon-utilizing Micro-organisms(Part 2) - On the Production of Single Cell Protein from Petroleum hydrocarbon with a yeast strain - (석유 탄화수소 이용 미생물에 관한 연구 (제 2 보) - 효모를 이용한 석유탄화수소로 부터 단백질 생산에 관하여 -)

  • Lee, Ke-Ho;Shin, Hyun-Kyung
    • Applied Biological Chemistry
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    • v.14 no.1
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    • pp.9-18
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    • 1971
  • In order to obtain basic information on the production of single cell protein from petroleum, more than 400 yeast strains were isolated from various soil samples in Korea utilizing petroleum hydrocarbon as the sole carbon source. A yeast strain showing the highest cell yield among the isolated strains was selected and identified. The optimal culture condition was searched in the flasks shaken throughout the procedure. And the growing characteristics for the selected yeast strain and chemical analysis of the yeast cell component were carried out. The results obtained were as follows: 1. The selected yeast strain was identified as Candida curvata and we named it Candida curvata-SNU 70. 2. The composition of the medium proposed for the present yeast strain is: Light Gas Oil 30ml, Urea 400mg, Ammonium sulfate 100mg, Potasium phosphate (monobasic) 670mg, Sodium phosphate (dibasic) 330mg, Magnesium sulfate 500mg, Calcium carbonate 3g, Yeast extract 50mg, Tween 20 0.05ml, Tap water 1,000ml. 3. Other culture conditions employed for the yeast were pH 5.5-7.0, temp. $30^{\circ}C$ under an affluent aerobic state. 4. Addition of light gas oil in portions to the culture media as the growth proceeded was more effective, especially in the cultivation on the higher oil concentration media. 5. Studies on the propagation of the yeast cells in the light gas oil medium revealed that the yeast has the lag phase lasted 16 hours and the logarithmic growth phase covered 16 to 28 hours. The specific growth rate was about $0.22\;hr^{-1}$ and doubling time was 3.2 hrs. during the logarithmic growth phase. 6. Under the cultural condition employed, the cell yield against the amount of light gas oil (wt%) was 16.1% and the protein content of the dried yeast cells was 48.4%.

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