• Journal of Animal Reproduction and Biotechnology
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• v.37 no.4
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• pp.292-297
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• 2022

Direct injection of CRISPR/Cas9 into zygotes enables the production of genetically modified nonhuman primates (NHPs) essential for modeling specific human diseases, such as Usher syndrome, and for developing novel therapeutic strategies. Usher syndrome is a rare genetic disease that causes loss of hearing, retinal degeneration, and problems with balance, and is attributed to a mutation in MYO7A, a gene that encodes an uncommon myosin motor protein expressed in the inner ear and retinal photoreceptors. To produce an Usher syndrome type 1B (USH1B) rhesus macaque model, we disrupted the MYO7A gene in developing zygotes. Identification of appropriately edited MYO7A embryos for knockout embryo transfer requires sequence analysis of material recovered from a trophectoderm (TE) cell biopsy. However, the TE biopsy procedure is labor intensive and could adversely impact embryo development. Recent studies have reported using cell-free DNA (cfDNA) from embryo culture media to detect aneuploid embryos in human in vitro fertilization (IVF) clinics. The cfDNA is released from the embryo during cell division or cell death, suggesting that cfDNA may be a viable resource for sequence analysis. Moreover, cfDNA collection is not invasive to the embryo and does not require special tools or expertise. We hypothesized that selection of appropriate edited embryos could be performed by analyzing cfDNA for MYO7A editing in embryo culture medium, and that this method would be advantageous for the subsequent generation of genetically modified NHPs. The purpose of this experiment is to determine whether cfDNA can be used to identify the target gene mutation of CRISPR/Cas9 injected embryos. In this study, we were able to obtain and utilize cfDNA to confirm the mutagenesis of MYO7A, but the method will require further optimization to obtain better accuracy before it can replace the TE biopsy approach.

• Korean journal of applied entomology
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• v.24 no.1 s.62
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• pp.45-50
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• 1985

The use of a modified procedure for the isolation of extrachromosomal DNA of low to high molecular weight, followed by agarose gel electrophoresis of the crude lysates, provided a simple screening procedure for detecting plasmids ranging in molecular weights from approximately 1 to more than 135 megadaltons from serovars of Bacillus thuringiensis. The procedure provides for a relatively large-volume stable lysate for isolation of plasmids for restriction endonuclease mapping and cloning procedures. The method was used for screening of plasm ids in 6 differenentially effective serovars of B. thuringiensis toxic to dipteran and lepidopteran insects. Relatively large plasmid DNAs of masses above 50 megadaltons (Mdal) were isolated from all of the serovars examined using this technique. The number of extrachromosomal DNAs detected in serovars of B. thuringiensis was 8 for israelensis, 10 for kurstaki, 13 for aizawai, 2 for dendrolimus, 1 for finitimus, and 6 for yunnanensis. Smaller plasmid DNAs were isolated in four of the six serovars that ranged in mass down to approximately 2 Mdal.

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.30-30
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• 2022

Philippines is the second world supplier of coconut by-products. As its first major genomics project, the Philippine Genome Center program for Agriculture (PGC-Agriculture) took the challenge to sequence and assemble the whole coconut genome. The project aims to provide advance genetics tools for our collaborating coconut researchers while taking the opportunity to initiate local capacity. Combination of different NGS platforms was explored and the Philippine 'Catigan Green Dwarf' (CATD) variety was selected with the breeders to be the crop's reference genome. A high quality genome assembly of CATD was generated and used to characterize important genes of coconut towards the development of resilient and outstanding varieties especially for added high-value traits. The talk will present the significant results of the project as published in various papers including the first report of whole genome sequence of a dwarf coconut variety. Updates will include the challenges hurdled and specific applications such as gene mining for host insect resistance and screening for least damaged coconuts (thus potentially insect resistant varieties). Genome-wide DNA markers as published and genes related to coconut oil qualitative/quantitative traits will also be presented, including initial molecular/biochemical studies that support nutritional and medicinal claims. A web-based genome database is currently built for ease access and wider utility of these genomics tools. Indeed, a major milestone accomplished by the coconut genomics research team, which was facilitated with the all-out government support and strong collaboration among multidisciplinary experts and partnership with advance research institutes.

• Korean Journal of Radiology
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• v.21 no.6
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• pp.736-745
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• 2020

Objective: To identify the initial chest computed tomography (CT) findings and clinical characteristics associated with the course of coronavirus disease 2019 (COVID-19) pneumonia. Materials and Methods: Baseline CT scans and clinical and laboratory data of 72 patients admitted with COVID-19 pneumonia (39 men, 46.2 ± 15.9 years) were retrospectively analyzed. Baseline CT findings including lobar distribution, presence of ground glass opacities, consolidation, linear opacities, and lung severity score were evaluated. The outcome event was recovery with hospital discharge. The time from symptom onset to discharge or the end of follow-up (for those remained hospitalized) was recorded. Data were censored in events such as death or discharge without recovery. Multivariable Cox proportional hazard regression was used to explore the association between initial CT, clinical or laboratory findings, and discharge with recovery, whereby hazard ratio (HR) values < 1 indicated a lower rate of discharge at four weeks and longer time until discharge. Results: Thirty-two patients recovered and were discharged during the study period with a median length of admission of 16 days (range, 9 to 25 days), while the rest remained hospitalized at the end of this study (median, 17.5 days; range, 4 to 27 days). None died during the study period. After controlling for age, onset time, lesion characteristics, number of lung lobes affected, and bilateral involvement, the lung severity score on baseline CT (> 4 vs. ≤ 4 [reference]: adjusted HR = 0.41 [95% confidence interval, CI = 0.18-0.92], p = 0.031) and initial lymphocyte count (reduced vs. normal or elevated [reference]: adjusted HR = 0.14 [95% CI = 0.03-0.60], p = 0.008) were two significant independent factors that influenced recovery and discharge. Conclusion: Lung severity score > 4 and reduced lymphocyte count at initial evaluation were independently associated with a significantly lower rate of recovery and discharge and extended hospitalization in patients admitted for COVID-19 pneumonia.

• Journal of Trauma and Injury
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• v.37 no.1
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• pp.48-59
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• 2024

Purpose: Research on rib fracture management has exponentially increased. Predicting fracture patterns based on the mechanism of injury (MOI) and other possible correlations may improve resource allocation and injury prevention strategies. The Chest Injury International Database (CIID) is the largest prospective repository of the operative and nonoperative management of patients with severe chest wall trauma. The purpose of this study was to determine whether the MOI is associated with the resulting rib fracture patterns. We hypothesized that specific MOIs would be associated with distinct rib fracture patterns. Methods: The CIID was queried to analyze fracture patterns based on the MOI. Patients were stratified by MOI: falls, motor vehicle collisions (MVCs), motorcycle collisions (MCCs), automobile-pedestrian collisions, and bicycle collisions. Fracture locations, associated injuries, and patient-specific variables were recorded. Heat maps were created to display the fracture incidence by rib location. Results: The study cohort consisted of 1,121 patients with a median RibScore of 2 (range, 0-3) and 9,353 fractures. The average age was 57±20 years, and 64% of patients were male. By MOI, the number of patients and fractures were as follows: falls (474 patients, 3,360 fractures), MVCs (353 patients, 3,268 fractures), MCCs (165 patients, 1,505 fractures), automobile-pedestrian collisions (70 patients, 713 fractures), and bicycle collisions (59 patients, 507 fractures). The most commonly injured rib was the sixth rib, and the most common fracture location was lateral. Statistically significant differences in the location and patterns of fractures were identified comparing each MOI, except for MCCs versus bicycle collisions. Conclusions: Different mechanisms of injury result in distinct rib fracture patterns. These different patterns should be considered in the workup and management of patients with thoracic injuries. Given these significant differences, future studies should account for both fracture location and the MOI to better define what populations benefit from surgical versus nonoperative management.

• Journal of Intelligence and Information Systems
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• v.17 no.2
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• pp.97-107
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• 2011

Nowadays there are a lot of issues about copyright infringement in the Internet world because the digital content on the network can be copied and delivered easily. Indeed the copied version has same quality with the original one. So, copyright owners and content provider want a powerful solution to protect their content. The popular one of the solutions was DRM (digital rights management) that is based on encryption technology and rights control. However, DRM-free service was launched after Steve Jobs who is CEO of Apple proposed a new music service paradigm without DRM, and the DRM is disappeared at the online music market. Even though the online music service decided to not equip the DRM solution, copyright owners and content providers are still searching a solution to protect their content. A solution to replace the DRM technology is digital audio watermarking technology which can embed copyright information into the music. In this paper, the author proposed a new audio watermarking algorithm with two approaches. First, the watermark information is generated by two dimensional barcode which has error correction code. So, the information can be recovered by itself if the errors fall into the range of the error tolerance. The other one is to use chirp sequence of CDMA (code division multiple access). These make the algorithm robust to the several malicious attacks. There are many 2D barcodes. Especially, QR code which is one of the matrix barcodes can express the information and the expression is freer than that of the other matrix barcodes. QR code has the square patterns with double at the three corners and these indicate the boundary of the symbol. This feature of the QR code is proper to express the watermark information. That is, because the QR code is 2D barcodes, nonlinear code and matrix code, it can be modulated to the spread spectrum and can be used for the watermarking algorithm. The proposed algorithm assigns the different spread spectrum sequences to the individual users respectively. In the case that the assigned code sequences are orthogonal, we can identify the watermark information of the individual user from an audio content. The algorithm used the Walsh code as an orthogonal code. The watermark information is rearranged to the 1D sequence from 2D barcode and modulated by the Walsh code. The modulated watermark information is embedded into the DCT (discrete cosine transform) domain of the original audio content. For the performance evaluation, I used 3 audio samples, "Amazing Grace", "Oh! Carol" and "Take me home country roads", The attacks for the robustness test were MP3 compression, echo attack, and sub woofer boost. The MP3 compression was performed by a tool of Cool Edit Pro 2.0. The specification of MP3 was CBR(Constant Bit Rate) 128kbps, 44,100Hz, and stereo. The echo attack had the echo with initial volume 70%, decay 75%, and delay 100msec. The sub woofer boost attack was a modification attack of low frequency part in the Fourier coefficients. The test results showed the proposed algorithm is robust to the attacks. In the MP3 attack, the strength of the watermark information is not affected, and then the watermark can be detected from all of the sample audios. In the sub woofer boost attack, the watermark was detected when the strength is 0.3. Also, in the case of echo attack, the watermark can be identified if the strength is greater and equal than 0.5.