• Journal of Life Science
• /
• v.30 no.3
• /
• pp.313-319
• /
• 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.

• The Journal of Korean Academy of Prosthodontics
• /
• v.55 no.4
• /
• pp.361-371
• /
• 2017

Purpose: We aimed to investigate the gene expression of human gingival fibroblasts on microgroove surface using DNA microarray. Materials and methods: Microgrooves were applied on grade II titanium discs to have 0/$0{\mu}m$ (NE0, control group), 60/$10{\mu}m$ (E60/10, experimental group) of respective width/depth by photolithography. The entire surface of the microgrooved Ti substrata was further acid etched and used as the two experimental groups in this study. Human gingival fibroblasts were cultured in the experimental group and the control group, and total RNA was extracted. The oligonucleotide microarray was performed to confirm the changes of various gene expression levels between experimental group and control group. Changes of gene expression level were determined at the pathway level by mapping the expression results of DNA chips, using the KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis. Results: Gene expression levels on E60/10 and NE0 were analyzed, there were 123 genes showing significant differences in expression more than 1.5 times on E60/10 microgrooved surface compared to NE0 surface, and 19 genes showing significant differences in expression more than 2 times. The KEGG pathway analysis confirmed the changes in gene expression levels under experimental conditions. Cell signaling, proliferation, and activity among the various gene expression results were identified. Conclusion: Microgrooved surfaces induce gene expression changes and related cell signaling. According to the results of this study, microgrooves can be used as the surface of various biomaterials which need to improve cell activity through gene expression changes and activation of cell signaling.

• Investigative Magnetic Resonance Imaging
• /
• v.9 no.1
• /
• pp.9-15
• /
• 2005

Purpose : This study attempted to investigate the effects of supply of highly concentrated $(30\%)$ oxygen on human ability of visuospatial cognition using time course data analysis of functional Magnetic Resonance Imaging (fMRI). Materials and Methods : To select an item set in the visuospatial performance test, two questionnaires with similar difficulty were developed through group testing. A group test was administered to 263 college students. Two types of questionnaire containing 20 questions were developed to measure the ability of visuospatial cognition. Eight college students (right-handed male, average age of 23.5 yrs) were examined for fMRI study. The experiment consisted of two runs of the visuospatial cognition testing, one with $21\%$ level of oxygen and the other with $30\%$ oxygen level. Each run consisted of 4 blocks, each containing control and visuospatial items. Functional brain images were taken from 37 MRI using the single-shot EPI method. Using the subtraction procedure, activated areas in the brain during visuospatial tasks were color-coded by t-score. To investigate the time course data in each activated area from brain images, 4 typical regions (cerebellum, occipital lobe, parietal lobe, and frontal lobe) were selected. Results : The average accuracy was $50.63{\pm}8.63$ and $62.50{\pm}9.64$ for $21\%\;and\;30\%$ oxygen respectively, and a statistically significant difference was found in the accuracy between the two types of oxygen (p<0.05). There were more activation areas observed at the cerebellum, occipital lobe, parietal lobe and frontal lobe with $30\%$ oxygen administration. The rate of increase in the cerebellum, occipital lobe and parietal lobe was $17\%$ and that of the frontal lobe, $50\%$. Especially, there were increase of intensity of BOLD signal at the parietal lobe with $30\%$ oxygen administration. The increase rate of the left parietal lobe was $1.4\%$ and that of the right parietal lobe, $1.7\%$. Conclusion : It is concluded that while performing visuospatial tasks, high concentrations of oxygen administration make oxygen administration sufficient, thus making neural network activate more, and the ability to perform visuospatial tasks increase.

• Journal of Biomedical Engineering Research
• /
• v.23 no.1
• /
• pp.9-16
• /
• 2002

Localized muscle fatigue can be identified by a downward shift of the EMG frequency typically represented by a fall in the median frequency The Present experimental study was Performed to investigate the time change of the median frequency and the muscle torque during maximal isometric back extension exercises at different exercise angles (0$^{\circ}$, 12$^{\circ}$, 36$^{\circ}$and 72$^{\circ}$) Twenty heath subjects (mean age : 24.35 $\pm$ 2.70) were Participated in this study Median frequency was extracted from EMG signals by employing the fast Fourier transform. Initial median frequency and the slope of median frequency was not significantly correlated with the muscle torque. Pearson's Product moment correlation was used to quantify the relationship between slopes of median frequency and torque. The results may suggest that the exorcise angle during maximal isometric back extension exercises does not affect the slopes of the median frequency and torque, and y-intercept of the median frequency among exercise angles There was no significant correlation between slopes of median frequency and torque. But there was a moderate correlation between median frequency and torque at each exercise angle. In conclusion, the exercise angle during maximal isometric back extension exercise is not a direct effect on slopes of median frequency and torque. But results showed that the shift of median frequency and torque shift were highly correlated in all subjects.

• Korean Journal of Biological Psychiatry
• /
• v.13 no.4
• /
• pp.279-288
• /
• 2006

Objectives : We investigated the association of SNP A and P1763 polymorphisms on dystrobrevin binding protein 1(DTNBP1) gene with smooth pursuit eye movement(SPEM) abnormality in Korean schizophrenic patients. Methods : We measured SPEM function in 217 Korean schizophrenics(male 116, female 101) and divided them into two groups, one is a good SPEM function group and the other is a poor SPEM function group. We then analyzed SNP A polymorphism and P1763 polymorphism on DTNBP1 gene from their DNAs extracted from their blood. We compared the differences of genotype and allele distributions of the two polymorphisms on DTNBP1 gene between the two groups. Results : The Ln S/N ratio(mean${\pm}$SD) of the good SPEM function group was $4.39{\pm}0.33$ and the ratio of poor SPEM function group was $3.17{\pm}0.71$. There were no statistically significant differences of age and male/female ratio between the two groups. There were no significant differences of genotype or allele distributions of the SNP A polymorphism and P1763 polymorphism on DTNBP1 gene between the two schizophrenic groups divided by SPEM function. Conclusion : The results suggest that SNP A polymorphism and P1763 polymorphism on DTNBP1 gene might not be related to SPEM function abnormality in schizophrenia.

• KSCE Journal of Civil and Environmental Engineering Research
• /
• v.30 no.3A
• /
• pp.297-307
• /
• 2010

Recently, maintenance engineering and technology for civil and building structures have begun to draw big attention and actually the number of structures that need to be evaluate on structural safety due to deterioration and performance degradation of structures are rapidly increasing. When stiffness is decreased because of deterioration of structures and member cracks, dynamic characteristics of structures would be changed. And it is important that the damaged areas and extent of the damage are correctly evaluated by analyzing dynamic characteristics from the actual behavior of a structure. In general, typical measurement instruments used for structure monitoring are dynamic instruments. Existing dynamic instruments are not easy to obtain reliable data when the cable connecting measurement sensors and device is long, and have uneconomical for 1 to 1 connection process between each sensor and instrument. Therefore, a method without attaching sensors to measure vibration at a long range is required. The representative applicable non-contact methods to measure the vibration of structures are laser doppler effect, a method using GPS, and image processing technique. The method using laser doppler effect shows relatively high accuracy but uneconomical while the method using GPS requires expensive equipment, and has its signal's own error and limited speed of sampling rate. But the method using image signal is simple and economical, and is proper to get vibration of inaccessible structures and dynamic characteristics. Image signals of camera instead of sensors had been recently used by many researchers. But the existing method, which records a point of a target attached on a structure and then measures vibration using image processing technique, could have relatively the limited objects of measurement. Therefore, this study conducted shaking table test and field load test to verify the validity of the method that can measure multi-point displacement responses of structures using image processing technique.

• Investigative Magnetic Resonance Imaging
• /
• v.8 no.2
• /
• pp.79-85
• /
• 2004

Purpose : To evaluate the usefulness and reproducibility of $^1H$ MRS in different 1.5 T MR machines with different coils to compare the SNR, scan time and the spectral patterns in different brain regions in normal volunteers. Materials and Methods : Localized $^1H$ MR spectroscopy ($^1H$ MRS) was performed in a total of 10 normal volunteers (age; 20-45 years) with spectral parameters adjusted by the autoprescan routine (PROBE package). In all volunteers, MRS was performed in a three times using conventional MRS (Signa Horizon) with 1 channel coil and upgraded MRS (Echospeed plus with EXCITE) with both 1 channel and 8 channel coil. Using these three different machines and coils, SNRs of the spectra in both phantom and volunteers and (pre)scan time of MRS were compared. Two regions of the human brain (basal ganglia and deep white matter) were examined and relative metabolite ratios (NAA/Cr, Cho/Cr, and mI/Cr ratios) were measured in all volunteers. For all spectra, a STEAM localization sequence with three-pulse CHESS $H_2O$ suppression was used, with the following acquisition parameters: TR=3.0/2.0 sec, TE=30 msec, TM=13.7 msec, SW=2500 Hz, SI=2048 pts, AVG : 64/128, and NEX=2/8 (Signa/Echospeed). Results : The SNR was about over $30\%$ higher in Echospeed machine and time for prescan and scan was almost same in different machines and coils. Reliable spectra were obtained on both MRS systems and there were no significant differences in spectral patterns and relative metabolite ratios in two brain regions (p>0.05). Conclusion : Both conventional and new MRI systems are highly reliable and reproducible for $^1H$ MR spectroscopic examinations in human brains and there are no significant differences in applications for $^1H$ MRS between two different MRI systems.

• Radiation Oncology Journal
• /
• v.21 no.1
• /
• pp.54-65
• /
• 2003

Purpose : To analyze the gene expression Profiles of uterine ceulcal cancer, and its variation after radiation therapy, with or without concurrent chemotherapy, using a CDNA microarray. Materials and Methods :Sixteen patients, 8 with squamous ceil carcinomas of the uterine cervix, who were treated with radiation alone, and the other 8 treated w14h concurrent chemo-radiation, were Included in the study. Before the starling of the treatment, tumor biopsies were carried out, and the second time biopsies were peformed after a radiation dose of 16.2$\~$27 Gy. Three normal cervix tissues were used as a control group. The microarray experiments were peformed with 5 groups of the total RNAs extracted individually and then admixed as control, pre-radiation therapy alone, during-radiation therapy alone, pre-chemoradiation therapy, and during-chemoradlation therapy. The 33P-iabeled CDNAS were synthesized from the total RNAs of each group, by reverse transcription, and then they were hybridized to the CDNA microarray membrane. The gene expression of each microarrays was captured by the intensity of each spot produced by the radioactive isotopes. The pixels per spot were counted with an Arrayguage, and were exported to Microsoft Excel The data were normalized by the Z transformation, and the comparisons were peformed on the Z-ratio values calculated. Results : The expressions of 15 genes, including integrin linked kinase (ILK), CDC28 protein kinase 2, Spry 2, and ERK 3, were increased with the Z-ratio values of over 2.0 for the cervix cancer tissues compared to those for the normal controls. Those genes were involved In cell growth and proliferation, cell cycle control, or signal transduction. The expressions of the other 6 genes, Including G protein coupled receptor kinase 5, were decreased with the Z-ratio values of below -2.0. After the radiation thorapy, most of the genes, with a previously Increase expressions, represented the decreased expression profiles, and the genes, with the Z-ratio values of over 2.0, were cyclic nucleotlde gated channel and 3 Expressed sequence tags (EST). In the concurrent chemo-radiation group, the genes involved in cell growth and proliferation, cell cycle control, and signal transduction were shown to have increased expressions compared to the radiation therapy alone group. The expressions of genes involved in anglogenesis (angiopoietln-2), immune reactions (formyl peptide receptor-iike 1), and DNA repair (CAMP phosphodiesterase) were increased, however, the expression of gene involved In apoptosls (death associated protein kinase) was decreased. Conclusion : The different kinds of genes involved in the development and progression of cervical cancer were identified with the CDNA microarray, and the proposed theory is that the proliferation signal stalls with ILK, and is amplified with Spry 2 and MAPK signaling, and the cellular mitoses are Increased with the increased expression oi Cdc 2 and cell division kinases. After the radiation therapy, the expression profiles demonstrated 4he evidence of the decreased cancer cell proliferation. There was no sigificant difference in the morphological findings of cell death between the radiation therapy aione and the chemo-radiation groups In the second time biopsy specimen, however, the gene expression profiles were markedly different, and the mechanism at the molecular level needs further study.

• Journal of Life Science
• /
• v.28 no.3
• /
• pp.339-344
• /
• 2018

Schizandra chinensis has been used as a traditional Chinese medicine and is known to have various bioactive components, including schizandrin and gomisin A. In the current study, we investigated the anti-inflammatory activities and their working mechanisms of ethanol extracts of pomace of Schizandra chinensis (PSC), schizandrin (SZ), and gomisin A (GA). First, we analyzed the effects of PSC on nitric oxide (NO) production and cell viabilities in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. The results indicated that PSC dramatically reduced NO production in LPS-activated RAW264.7 cells in a dose-dependent manner without affecting cell viabilities. PSC also decreased the expression of pro-inflammatory genes iNOS and COX-2, whereas the expression of TNF-${\alpha}$ was not affected by PSC. In addition, PSC inhibited phosphorylation of p38, ERK1/2, and JNK but did not change the expression of their total protein. The results indicate that PSC can regulate LPS-induced inflammatory responses by suppressing MAPK (mitogen-activated protein kinase) signaling. We also analyzed the effects of SZ and GA on NO production and cell viabilities in RAW264.7 cells. The results showed that SZ and GA also decreased NO production in a dose-dependent manner in LPS-activated RAW 264.7 cells without affecting cell viabilities. SZ reduced the expression of iNOS, whereas GA downregulated iNOS and COX-2. Overall, these findings clarify the molecular mechanisms of the anti-inflammatory effects mediated by PSC, SZ, and GA.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.43 no.2
• /
• pp.224-230
• /
• 2014

Marine algae have long been recognized as a health and beauty food, based on its anti-tumor, anti-inflammatory and anti-obesity activities. In this study, methanol extracts were prepared from 10 different phaeophyta, after which DPPH radical scavenging and cytoprotective activities of HT-22 cells against ${\beta}$-amyloid protein ($A{\beta}$), which has neurotoxic effects, were investigated. In DPPH experiments, Ecklonia cava and Ishige okamurai showed strong ROS scavenging activities, whereas eight other phaeophyta including Petalonia binghamiae (P. bin) showed weak ROS scavenging activities. To validate the cytoprotective effects of 10 different phaeophyta in $A{\beta}$-induced HT-22 cells, protein expression levels of APP, BACE1, iNOS, phosphorylated ERK1/2, phosphorylated p38 and phosphorylated JNK1/2 were determined along with MTT assay. In the MTT assay, P. bin showed the best effective cytoprotective activity at a concentrations of $25{\mu}g/mL$, whereas Sargassum confusum, Colpomenia sinuosa, Myelophycus simplex, and Sargassum hemiphyllum showed potential. Determination of protein expression levels related to $A{\beta}$-induced neurotoxicity in the five selected phaeophyta showed that P. bin inhibited BACE1 and iNOS expression in $A{\beta}$-induced HT-22 cells. These results indicate that the cytoprotective effects of P. bin are mediated by suppressing the pathways involving $A{\beta}$-induced ERK and p38 activation.