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Application of the CRISPR/Cas System for Point-of-care Diagnosis of Cattle Disease (현장에서 가축질병을 진단하기 위한 CRISPR/Cas 시스템의 활용)

  • Lee, Wonhee;Lee, Yoonseok
    • Journal of Life Science
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    • v.30 no.3
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    • pp.313-319
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    • 2020
  • Recently, cattle epidemic diseases are caused by a pathogen such as a virus or bacterium. Such diseases can spread through various pathways, such as feed intake, respiration, and contact between livestock. Diagnosis based on the ELISA (Enzyme-linked immunosorbent assay) and PCR (Polymerase chain reaction) methods has limitations because these traditional diagnostic methods are time consuming assays that require multiple steps and dedicated equipment. In this review, we propose the use of the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) Cas system based on DNA and RNA levels for early point-of-care diagnosis in cattle. In the CRISPR/Cas system, Cas effectors are classified into two classes and six subtypes. The Cas effectors included in class 2 are typically Cas9 in type II, Cas12 in type V (Cas12a and Cas12b) and Cas13 in type VI (Cas13a and Cas13b). The CRISPR/Cas system uses reporter molecules that are attached to the ssDNA strands. When the Cas enzyme cuts the ssDNA, these reporters either fluoresce or change color, indicating the presence of a specific disease marker. There are several steps in the development of a CRISPR/Cas system. The first is to select the Cas enzyme depending on DNA or RNA from pathogens (viruses or bacteria). Based on that, the next step is to integrate the optimal amplification, transducing method, and signal reporter. The CRISPR/Cas system is a powerful diagnostic tool using a gene-editing method, which is faster, better, and cheaper than traditional methods. This system could be used for early diagnosis of epidemic cattle diseases and help to control their spread.

Ablation Rate and Intrapulpal Temperature by Addition of Water Spray During Er:YAG Laser Irradiation (Er:YAG laser를 이용한 치아삭제시 물분사량이 삭제율과 치수내 온도변화에 미치는 영향)

  • Kim, Jung-Moon;Kim, Mee-Eun;Kim, Ki-Suk
    • Journal of Oral Medicine and Pain
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    • v.30 no.3
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    • pp.375-381
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    • 2005
  • Er:YAG laser has been considered a promising alternative to dental drill and many researches indicate that adjustment to variable parameters, including water flow rate, pulse energy and pulse repetition rate, can be made to improve ablation ability and efficiency of the laser. Of these parameters, addition of water spray during irradiation has been thought to ablate dental hard tissue more rapidly and safely. The purpose of this study was to investigate tooth ablation amount by Er:YAG laser irradiation as related to varied water flow rates added and, ultimately to find the most effective water flow rate for ablation. In addition, the temperature change of pulp chamber during irradiation was also monitored on the irradiated and opposite pulpal walls, respectively. An Er:YAG laser with contact mode was employed. Extracted human molars were split into two pieces for ablation experiment. Pulse energies of 200 and 300 mJ with a pulse repetition rate of 20 Hz and 5 water flow rates (1.6, 3.0, 5.0, 7.0, and 10.0 ml/min) were applied. Each irradiation was performed for 3 seconds. According to these parameters, experimental groups were divided into 10 subgroups which consisted of 5 specimens. For temperature experiment, another 5 tooth-specimens were prepared in the manner that pulp chamber was open through access cavity preparation and two temperature-measuring probes were placed respectively on the irradiated and the opposite walls of pulp chamber. From the experiment on ablation amount related to different water flow rates, it was shown that the least water flow rate of 1.6 ml/min ablated more than any other water flow rates (p<0.000). When the irradiation for 3 seconds, combined with the pulse repetition time of 20Hz and the water flow rate of 1.6 ml/min was done to tooth specimen, the temperature rise was not noticeable both on the irradiated and the opposite pulpal walls (less than 3$^{\circ}C$) and there was no significant difference in temperature rise between the two pulse energies, 200 and 300 mJ. From the results of this study, it is suggested that tooth ablation with Er:YAG laser can be done effectively and safely at a energy between 200 and 300 mJ/pulse and a pulse repetition rate of 20 Hz when the lasing is conjugated with the water flow rate of 1.6ml/min.

Evaluation of Efficacy and Development of Predictive Model of Sanitizers and Disinfectants on Reduction of Microorganisms on Food Contact Surfaces (스테인리스 스틸 식품기구 표면에 사용되는 주요 살균소독제의 살균력 평가 및 살균예측모델 개발)

  • Lee, Yu-Si;Ha, Sang-Do;Kim, Dong-Ho;Park, Joon-Hee
    • Journal of Food Hygiene and Safety
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    • v.26 no.3
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    • pp.203-208
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    • 2011
  • This study was to evaluate the efficacy of sanitizer concentrations and treatment time against two major toad-borne pathogenic microorganisms such as Escherichia coli and Staphylococcus aureus on a stainless steel surface. As a result, stainless steel, treated with 100 ppm of chlorine showed reduction of E. coli(1.56, 1.49, 1.95 log cfu/25 $cm^2$) and S. aureus(0.49, 0.88, 1.27 log cfu/25 $cm^2$) after 0, 5 and 10 min, but none was not detected in treatment with 200 ppm. The population of E. coli(0.73, 0.90, 1.55 log cfu/25 $cm^2$) and S. aureus(0.37, 1.00, 1.45 log cfu/25 $cm^2$) reduced in 35.5% ethanol treated group, but none was not detected in treatment with 70%. The population was reduced E coli(0.28, 0.64, 1.07 cfu/25 $cm^2$) and S. aureus(0.53, 0.87, 0.99 log cfu/25 $cm^2$) by treatment with 45.5 ppm of hydrogen peroxide, but none was not detected in treatment with 91 ppm. Quarternary ammonium compound with 100 ppm was reduced E. coli(0.82, 1.62, 1.71 log cfu/25 $cm^2$) and S. aureus(0.46, 0.93, 1.38 log cfu/25 $cm^2$), but none was not detected in treatment with 200 ppm. Predictive models of sterilization for all 4 disinfectants were suitable to use with $r^2$ value of higher than 0.94. These models may be of use to food services and manufacture of safe products by controlling E. coli and S. aureus without the need for further detection of the organisms.

Study for the Geochemical Reaction of Ca-feldspar, Amphibole and Olivine with Supercritical $CO_2$ and Brine on the $CO_2$ Sequestration Condition (이산화탄소 지중저장 조건에서 초임계이산화탄소와 염수 반응에 의한 Ca-사장석, 각섬석, 감람석의 지화학적 변화 연구)

  • Kang, Hyun-Min;Park, Min-Ho;Park, Sang-Hee;Lee, Min-Hee;Wang, Soo-Kyun
    • Economic and Environmental Geology
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    • v.44 no.2
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    • pp.123-133
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    • 2011
  • The lab scale experiments to investigate the geochemical reaction among supercritical $CO_2$-mineral-brine which occurs at $CO_2$ sequestration sites were performed. High pressurized cell system (l00 bar and $50^{\circ}C$) was designed to create supercritical $CO_2$ in the cell, simulating the sub-surface $CO_2$ storage site. From the high pressurized cell experiment, the surface changes of Ca-feldspar, amphibole (tremolite) and olivine, resulted from the supercritical $CO_2$-mineral-brine reaction, were observed and the dissolution of minerals into the brine was also investigated. The mineral slabs were polished and three locations on the surface were randomly selected for the image analysis of SPM and the surface roughness value (SRV) of those locations were calculated to quantify the change of mineral surface for 30 days. At a certain time interval, SPM images and SRVs of the same mineral surface were acquired. The secondary minerals precipitated on the mineral surfaces were also analyzed on SEM/EDS after the experiment. From the experiments, the average SRV of Ca-feldspar increased from 2.77 nm to 20.87 nm for 30 days, suggesting that the dissolution of Ca-feldspar occurs in active when the feldspars contact with supercritical $CO_2$ and brine. For the amphibole, the average SRV increased from 2.54 nm to 8.31 nm and for the olivine from 0.77 nm to 11.03 run. For the Ca-feldspar, $Ca^{2+}$, $Na^+$, $Fe^{2+}$, $Si^{4+}$, $K^+$ and $Mg^{2+}$ were dissolved in the highest order and $Si^{4+}$, $Ca^{2+}$, $Fe^{2+}$ and $Mg^{2+}$ for the amphibole. Fe (or Mg) - oxides were precipitated as the secondary minerals on the surfaces of amphibole and olivine after 30 days reaction. Results suggested that $Ca^{2+}$, $Fe^{2+}$ and $Mg^{2+}$ rich minerals would be significantly weathered when it contacts with the supercritical $CO_2$ and brine at $CO_2$ sequestration sites.

3-Dimensional Reconstruction of Parallel fiber-Purkinje Cell Synapses Using High-Voltage Electron Microscopy (고압전자현미경을 이용한 소뇌 평행섬유-조롱박세포간 신경연접의 3차원 재구성)

  • Lee, Kea-Joo;Kweon, Hee-Seok;Kang, Ji-Seoun;Rhyu, Im-Joo
    • Applied Microscopy
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    • v.35 no.1
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    • pp.31-39
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    • 2005
  • Synapses are contact points where one neuron communicates with another. The morphological change of synapses under various physiological or pathological conditions has long been hypothesized to modify their functional properties. 3-dimensional (3-D) reconstruction of synapses with serial ultrathin sections has contributed to the understanding of ultrastructural dimensions and compositions of synapses. The 3-D reconstruction procedures, however, require a great amount of expertise as well as include prohibitively timeconsuming processes. Here, we introduce efficient 3-D reconstruction technique using high-voltage electron microscopy (HVEM). Primarily, we established an optimal section thickness and staining condition to observe synaptic structures in detail under HVEM. The result showed that synaptic profiles were preserved at the section thickness of 250 nm without the overlapping of synaptic ultrastructures. An increase in the reaction time of en bloc staining was most efficient to enhance contrast than the extension of postembedding staining or the addition of uranyl acetate during dehydration. Then, 3-D reconstruction of parallel fiber-Purkinje cell synapses in the rat cerebellum was carried out with serial HVEM images and reconstruction software. The images were aligned and the contours of synapses were outlined on each section. 3-D synapses were finally extracted from the section files by grouping all the synaptic contours. The reconstructed synapse model clearly demonstrated the configuration of pre and postsynaptic components. These results suggest that 3-D reconstruction of synapses using HVEM is much efficient and suitable for massive quantitative studies on synaptic connectivity than conventional TEM approach using numerous ultrathin sections.

Structural Safety Assessment of a Sunken Ship Considering Hull Corrosion and Damaged Members - Focus on the Sunken Ship 'No. 7 HaeSung' - (선체 부식 및 손상 부재를 고려한 침몰선박의 구조 안전성 평가에 관한 연구 - 제7 해성호를 중심으로 -)

  • Lee, Seung Hyun;Kim, Won Don;Suh, Jae-Joon
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.19 no.4
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    • pp.332-340
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    • 2016
  • Sunken ships cause damage to the environment due to the dispersal of fuel oil and harmful cargo goods in the hull. Since the sunken ship is mostly flooded by the seabed, it tends to be in a relatively stable condition. However, the heavy body, together with the load of remaining goods in the cargo hold, the constant contact with the seabed, and ocean currents and tidal waves, can affect dispersal of residual fuel oils out of the sunken ship. Corrosion of the sunken ship starts upon sinking, decreasing the thickness of the hull structure and sub-materials. Therefore, it is necessary to assess the structural stability against the potential breakdown of the sunken ship. Whilst evaluating the danger of the sunken ship, this result should be reflected in 'the possible discharge'. This study was undertaken to suggest a procedure for a step by step evaluation to assess the structural stability a sunken ship. The structural stability assessment to estimate the collapsibility of the hull was structure targeted at the sunken ship 'No. 7 HaeSung', which was classified as the prime example for the intensive management of sunken ships. This study was undertaken to suggest a procedure for a step by step evaluation to assess the structural stability a sunken ship and to propose a method to conduct a structural safety assessment that estimates the collapsibility of the hull by targeting the sunken ship 'No. 7 HaeSung',which was classified as the prime example for the intensive management of sunken ships. The collapsibility of the hull structure was estimated Based on the damage size of the hull structure, and the corrosion rate of the hull structure and sub-materials due to the seawater after sinking. It was confirmed that there was a low possibility of the total destruction of the hull structure at the current time. However, there is a high possibility in the potential failure of the hull structure due to increased rate of corrosion thereafter. Therefore, we believe continuous study on influence of corrosion and marine environment change to sunken ship's structural safety is necessary.

A study on the implementation of Medical Telemetry systems using wireless public data network (무선공중망을 이용한 의료 정보 데이터 원격 모니터링 시스템에 관한 연구)

  • 이택규;김영길
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2000.10a
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    • pp.278-283
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    • 2000
  • As information communication technology developed we could check our blood pressure, pulsation electrocardiogram, SpO2 and blood test easily at home. To check our health at ordinary times is able though interlocking the house medical instrument with the wireless public data network This service will help the inconvenience to visit the hospital everytime and will save the individual's time and cost. In each house an organism data which is detected from the human body will be transmitted to the distance hospital and will be essentially applied through wireless public data network The medical information transmit system is utilized by wireless close range network It would transmit the obtained organism signal wirelessly from the personal device to the main center system in the hospital. Remote telemetry system is embodied by utilizing wireless media access protocol. The protocol is embodied by grafting CSMA/CA(Carrier Sense Multiple Access with Collision Avoidance) protocol falling mode which is standards from IEEE 802.11. Among the house care telemetry system which could measure blood pressure, pulsation, electrocardiogram, SpO2 the study embodies the ECC(electrocardiograph) measure part. It within the ECC function into the movable device and add 900㎒ band wireless public data interface. Then the aged, the patients even anyone in the house could obtain ECG and keep, record the data. It would be essential to control those who had a health-examination heart diseases or more complicated heart diseases and to observe the latent heart disease patient continuously. To embody the medical information transmit system which is based on wireless network. It would transmit the ECG data among the organism signal data which would be utilized by wireless network modem and NCL(Native Control Language) protocol to contact through wireless network Through the SCR(Standard Context Routing) protocol in the network it will be connected to the wired host computer. The computer will check the recorded individual information and the obtained ECC data then send the correspond examination to the movable device. The study suggests the medical transmit system model utilized by the wireless public data network.

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Metamorphism and Deformation of the Late Paleozoic Pyeongan Supergroup in the Taebaeksan Basin: Reviews on the Permo-Triassic Songrim Orogeny (태백산분지에 분포하는 후기 고생대 평안누층군의 변성-변형작용: 페름-삼첩기 송림 조산운동의 고찰)

  • Kim, Hyeong-Soo
    • The Journal of the Petrological Society of Korea
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    • v.21 no.2
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    • pp.151-171
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    • 2012
  • The Permo-Triassic Songrim orogeny in the Korean peninsula was a major tectonic event involving complicated continental collisions at the eastern margin of Eurasia. Based on the previous studies on the metamorphic and deformations features of the Songrim orogeny, this paper presents metamorphic and structural characteristics and timing of the Songrim orogeny in the Taebaeksan basin, and discuss about correlation of the tectono-metamorphic evolution of the Taebaeksan basin with the Okcheon basin and the Imjingang belt with a combined analysis of bulk crustal shortening direction, metamorphic P-T and T-t (time) paths. The metapelites in the Pyeongan Supergroup in the northeastern margin of the Taebaeksan basin have experienced lower-temperature/medium-pressure (LT/MP) regional metamorphism followed by high-temperature contact metamorphism due to the Jurassic granite intrusion. The earlier LT/MP regional metamorphism produced two loops of clockwise P-T-d (deformation) paths combined with four deformation events ($D_1-D_4$). The first loop concomitant with $D_1$ and $D_2$ occurred at $400-500^{\circ}C$, 1.5-3.0 kbar, and related with growth of syn-$D_1$ chloritoid and andalusite, post-$D_1$ margarite, Ca-rich syn-$D_2$ or post-$D_2$ plagioclase. The second loop accompanying $D_3$ and $D_4$ occurred at $520-580^{\circ}C$, 2.0-6.0 kbar, and associated with the growth of syn-$D_3$ garnet and staurolite, and syn-$D_4$ and/or post-$D_4$ andalusite porphyroblasts. Furthermore the syn-$D_1$ chloritoid and andalusite porphyroblasts grew during E-W bulk crustal shortening, whereas the syn-$D_3$ garnet and staurolite, and the syn-$D_4$ and/or post-$D_4$ andalusite porphyroblasts have grown under N-S bulk crustal shortening. The similarity in the characteristics and timing of the metamorphism and bulk crustal shortening directions between the Okcheon and Imjingang belts suggest that the peak metamorphic conditions tend to increase toward the western part (Imjingang belt and southwestern part of the Gyeonggi Massif) from the eastern part (Taebaeksan basin). The E-W bulk crustal shortening influenced the eastern part of the Okcheon belt, whereas the N-S bulk crustal shortening resulted in strong deformation in the Imjingang and Okcheon belts. Consequently, the Permo-Triassic Songrim orogeny in the Korean peninsula is probably not only related to collision of the North and South China blocks, but also to the amalgamation of terrane fragments at the eastern Eurasia margin (e.g., collision of the Sino-Korean continent and the Hida-Oki terrane).

The Effects of Calcium-Phosphate Coated Xenogenic Bone and Type I Collagen for Bone Regeneration on the Calvarial Defects in Rabbits (Ca-P 박막 이종골과 제 1형 교원질이 토끼 두개골 결손부의 골재생에 미치는 영향)

  • Kim, Chang-Han;Park, Jin-Woo;Lee, Jae-Mok;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.34 no.1
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    • pp.223-241
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    • 2004
  • The purpose of this present study evaluated the osseous response around Ca-P coated xenogenic bone and compared osteogenic potential of Ca-P coated xenogenic bone to that of combination with type I collagen derived from bovine tendon as a biocompatible binder to prevent migration of bone particle on the repair of calvarial defects in rabbits. To study the effects of Ca-P coated xenogenic bone and collagen on bone healing, four 5-mm-diameter skull defect were made in calvaria with trephine filled with an autogenous bone chip or Ca-P coated xenogenic bone or Ca-P coated xenogenic bone and type I collagen (1:1 mixture by volume) or left empty. The defects were evaluated histologically at 1, 2, 4 and 8 weeks following implantation. Ca-P coated xenogenic bone at the calvarial defects of rabbits showed osteoconductivity at the margin of defect in the early stage of bony healing, but no direct contact with new bone was observed. With time passed by, it was resorbed slowly and showed consistent inflammatory reaction. An additional use of type I collagen derived from bovine tendon improved clinical handling, but no new bone formation was observed histologically. Above all, autogenous bone graft showed most prominent healing in quantity and density of new bone formation. According to this study, the use of Ca-P coated xenogenic bone alone and combination with type I collagen did not showed effective healing in quantity and density of new bone formation.

The Effect of N-acetyl cysteine (NAC) loading on the bone formation surrounding sandblasted and large-grit and acid-etched implants in the dog: A pilot study (성견에 식립된 sandblasted and large-grit and acid-etched 임플란트에서 N-acetyl cysteine(NAC)의 탑재가 주위 골형성에 미치는 영향에 대한 선행연구)

  • Seo, Jae-Min;Kim, In-Ju;Bae, Min-Soo;Lee, Jung-Jin;Ahn, Seung-Geun
    • The Journal of Korean Academy of Prosthodontics
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    • v.58 no.2
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    • pp.77-85
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    • 2020
  • Purpose: The aim of the present study was to investigate the effects of N-acetyl cysteine (NAC) loading on the bone formation surrounding sandblasted, large-grit and acid-etched (SLA) implants. Materials and methods: Implantation of NAC loaded SLA implants (NSI group) and SLA implants (SI group) was performed bilaterally in the mandible of 4 adult beagle dogs (each group, n = 8). The animals were sacrificed after a healing period of 3 and 6 weeks, respectively (n = 2 animal each). Dissected blocks were processed for histomorphometrical analysis. Bone to implant contact percentage (BIC%) and bone volume (BV%) were assessed histomorphometrically. Results: BIC% of NAC loaded SLA implants were about 50% higher than that of SLA implants at 3 weeks of bone healing, but not significantly (51.79 vs 35.43%; P=.185). BV% of NAC loaded SLA implants were significantly higher than that of SLA implants at 3 weeks of bone healing (45.09 vs 37.57 %; P=.044). At 6 weeks of bone healing, BIC% and BV% of two experimental groups were similar (P>.05). Conclusion: Within the limits of the present study, NAC loading have a positive effects on the early bone formation surrounding SLA implants. So, it might be concluded that NAC loading enhance the osseointegration and shorten the healing time after implantation of the SLA implants.