• Title/Summary/Keyword: Cycle threshold value

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Diapause hormone of the silkworm, Bombyx mori : Structure and function

  • Okitsugu Yamashita
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 1997.06a
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    • pp.51-72
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    • 1997
  • Diapause hormone (DH) is a neuropeptide hormone which is secreted from the suboesophageal ganglion (SG) and is responsible for induction of embryonic diapause of the silkworm, Bombyx mori. DH is isolated from SGs and determined to be a 24 amino acid peptide amide. The cDNA encodes the polyprotein precursor from which DH, pheromone biosynthesis activating neuropeptide (PBAN) and three other neuropeptides are released and become matured. The C-terminal FXPRL-NH2 sequence of DH is essential but not sufficient for expression of full activity. Recently, we have isolated a unique hydrohobic peptide (VAP peptide) with a slight diapause egg induceing activity from organic solvent extracts of the male adult heads of the silkworm. The VAP peptide itself has no diapause inducing activity, but enhances DH activity through reducing ED50 value and the threshold concentration of DH. The DH-PBAN gene is composed of 6 exons interrupted by 5 introns and is expressed in 12 neurosecretory cells of the SG. The incubation of eggs at 25$^{\circ}C$, which induces embryonic diapause in the progeny, caused DH-PBAN mRNA content to increase at 5 different stages in the life cycle. By contrast, a 15$^{\circ}C$ incubation only induced expression of the gene at the late phrase adult stage. The temperature-controlled expression of DH-PBAN gene is closely correlated to the incidence of diapause, indicating that DH-PBAN gene expression is the initial event leading to diapause induction. DH acts to stimulate trehalase activity in developing ovary to bring about hyprglycogenism in mature eggs, a prerequisite metabolism for diapause initiation. Using in vivo and in vitro systems, DH is clearly shown to induce trehalase gene expression in developing ovaries. New protein synthesis is not needed for this process, but a Ca2+-dependent proteinkinase seems to be involved. Quite recently, we have sucessfully applied a new and potent trehalase inhibitor (Trehazoline) to reudce glycogen content in developing ovaries. The eggs deficient in glycogen were also able to enter diapause as the natural eggs do, so that we could provide the new egg system to reconsider the diapause associated metabolism other than the glycogen-sorbitol metabolic system.

Effect of Microstructural Factors on Fatigue and Fatigue Crack Propagation Behaviors of Mill-Annealed Ti-6Al-4V Alloy (미세조직적 인자가 밀소둔된 Ti-6Al-4V 합금의 피로 및 피로균열전파 거동에 미치는 영향)

  • Park, Sanghoo;Kim, Sumin;Lee, Daeun;Ahn, Soojin;Kim, Sangshik
    • Korean Journal of Metals and Materials
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    • v.56 no.12
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    • pp.845-853
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    • 2018
  • To understand the effect of microstructural factors (i.e., the size of ${\alpha}$ phase, equiaxed vs bimodal structure) on high cycle fatigue (HCF) and fatigue crack propagation (FCP) behaviors of mill-annealed Ti-6Al-4V (Ti64) alloy, three specimens of EQ (equiaxed)-8 (8 indicates the size of ${\alpha}$ grain), BM (bimodal)-8, and BM-16 were studied. The uniaxial HCF and FCP tests were conducted at an R ratio of 0.1 under sinusoidal fatigue loading. The microstructural influence (i.e., EQ vs BM) was not significant on the tensile properties of mill-annealed Ti64 alloy, and showed an increase in tensile strength and elongation with decreasing gauge thickness from 50 mm to 1.3 mm. The microstructure, on the other hand, affected the resistance to HCF substantially. It was found that the EQ structure in mill-annealed Ti64 has better resistance to HCF than the BM structure, as a result of different crack initiation mechanism. Unlike HCF behavior, the effect of microstructural features on the FCP behavior of mill-annealed Ti64 was not significant. Among the three specimens, BM-16 specimen showed the highest near-threshold ΔK value, probably because it had the greatest slip reversibility with large ${\alpha}$ grains. The effect of microstructural factors on the HCF and FCP behaviors of mill-annealed Ti64 alloy are discussed based on fractographic and micrographic observations.

Viral load and rebound in children with coronavirus disease 2019 during the first outbreak in Daegu city

  • Chu, Mi Ae;Jang, Yoon Young;Lee, Dong Won;Kim, Sung Hoon;Ryoo, Namhee;Park, Sunggyun;Lee, Jae Hee;Chung, Hai Lee
    • Clinical and Experimental Pediatrics
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    • v.64 no.12
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    • pp.652-660
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    • 2021
  • Background: Viral load and shedding duration are highly associated with the transmission of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. However, limited studies have reported on viral load or shedding in children and adolescents infected with sudden acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Purpose: This study aimed to investigate the natural course of viral load in asymptomatic or mild pediatric cases. Methods: Thirty-one children (<18 years) with confirmed SARS-CoV-2 infection were hospitalized and enrolled in this study. Viral loads were evaluated in nasopharyngeal swab samples using real-time reverse transcription polymerase chain reaction (E, RdRp, N genes). cycle threshold (Ct) values were measured when patients met the clinical criteria to be released from quarantine. Results: The mean age of the patients was 9.8 years, 18 (58%) had mild disease, and 13 (42%) were asymptomatic. Most children were infected by adult family members, most commonly by their mothers. The most common symptoms were fever and sputum (26%), followed by cough and runny nose. Nine patients (29%) had a high or intermediate viral load (Ct value≤30) when they had no clinical symptoms. Viral load showed no difference between symptomatic and asymptomatic patients. Viral rebounds were found in 15 cases (48%), which contributed to prolonged viral detection. The mean duration of viral detection was 25.6 days. Viral loads were significantly lower in patients with viral rebounds than in those with no rebound (E, P=0.003; RdRp, P=0.01; N, P=0.02). Conclusion: Our study showed that many pediatric patients with coronavirus disease 2019 (COVID-19) experienced viral rebound and showed viral detection for more than 3 weeks. Further studies are needed to investigate the relationship between viral rebound and infectiousness in COVID-19.

Analysis of receptor like kinase (RLK) gene to stress in rice (Oryza sativa L.) using real-time PCR (Real-time PCR을 이용한 스트레스에 따른 벼의 Receptor like kinase (RLK) 유전자의 발현 변화 분석)

  • Kang, Min-Hee;Kim, Il-Wook;Han, Sang-Hoon;Yun, Choong-Hyo;Yoon, Byoung-Su
    • Journal of Plant Biotechnology
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    • v.35 no.4
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    • pp.281-290
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    • 2008
  • In plant, Receptor-like kinases (RLKs) are protein family, though its function is not yet understood, consisted of a predicted signal sequence, single transmembrane region, and cytoplasmic kinase domain. RLKs are involved in hormonal response pathways, cell differentiation, plant growth and development, self-incompatibility, and symbiont and pathogen recognition. In this study, expression levels of RLG1, RLG5, RLG6, RLG#6, RLG8, RLG10, RLG17, RLG18 and RLG20 were analyzed by Real-time PCR, when rice (Oryzae sativa) was treated abiotic stress. The expression levels of all RLGs were compared each other by analyzed value of threshold cycles ($C_T$). Consequently, RLGs were suppressed by NaCl as salinity stress, and expression of each RLK genes were showed difference treated salicylic acid and wound, respectively. However, All RLGs were induced under low temperature condition. Therefore, our results indicate protection-function of RLK genes to be an early response of rice against cold weather.

Predictive value of sperm motility characteristics assessed by computer-assisted sperm analysis in intrauterine insemination with superovulation in couples with unexplained infertility

  • Youn, Joung-Sub;Cha, Sun-Hwa;Park, Chan-Woo;Yang, Kwang-Moon;Kim, Jin-Yeong;Koong, Mi-Kyoung;Kang, Inn-Soo;Song, In-Ok;Han, Sang-Chul
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.1
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    • pp.47-52
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    • 2011
  • Objective: To determine whether characteristics of sperm motility obtained by computer-assisted sperm analysis (CASA) could predict pregnancy after intrauterine insemination (IUI) in couples with unexplained infertility. Methods: Three hundred eighty-three cycles of intrauterine insemination with superovulation were retrospectively analyzed. Semen analysis was performed with CASA before and after swim-up and the parameters were compared between pregnant and non-pregnant women. Results: The pregnancy rate per cycle was 14.1%. Pregnant and non-pregnant women were comparable in terms of age, infertility duration, the number of dominant follicles. While sperm concentration, motility, and parameters such as average path velocity (VAP) and percentage rapid (RAPID) before semen preparation were significantly different between the pregnancy and non-pregnancy groups, there were no differences in sperm parameters when comparing the two groups after preparation. Using a receiver operating characteristic curve to measure sensitivity and specificity, the optimal threshold value for the predictors of pregnancy was revealed to be a concentration of ${\geq}111{\times}10^6/mL$, a motility of ${\geq}$ 51.4%, and RAPID ${\geq}$ 30.1% before preparation for IUI. Conclusion: Sperm parameters including concentration, motility, and RAPID before sperm preparation could have predictive value for pregnancy outcome after intrauterine insemination with superovulation in couples with unexplained infertility, and would be helpful when counseling patients before they make the decision to proceed with IVF/ICSI-ET.

Evaluation of Galactomannan Enzyme Immunoassay and Quantitative Real-Time PCR for the Diagnosis of Invasive Pulmonary Aspergillosis in a Rat Model

  • Lin, Jian-Cong;Xing, Yan-Li;Xu, Wen-Ming;Li, Ming;Bo, Pang;Niu, Yuan-Yuan;Zhang, Chang-Ran
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1044-1050
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    • 2014
  • Since there is no consensus about the most reliable assays to detect invasive aspergillosis from samples obtained by minimally invasive or noninvasive methods, we compared the efficacy of an enzyme-linked immunosorbent assay (ELISA) for galactomannan (GM) detection and quantitative real-time PCR assay (qRT-PCR) for the diagnosis of invasive pulmonary aspergillosis. Neutropenic, male Sprague-Dawley rats (specific pathogen free; 8 weeks old; weight, $200{\pm}20g$) were immunosuppressed with cyclophosphamide and infected with Aspergillus fumigatus intratracheally. Tissue and whole blood samples were harvested on days 1, 3, 5, and 7 post-infection and examined with GM ELISA and qRT-PCR. The A. fumigatus DNA detection sequence was detected in the following number of samples from 12 immunosuppressed, infected rats examined on the scheduled days: day 1 (0/12), day 3 (0/12), day 5 (6/12), and day 7 (8/12) post-infection. The sensitivity and specificity of the qRT-PCR assay was 29.2% and 100%, respectively. Receiver operating characteristic curve (ROC) analysis indicated a Ct (cycle threshold) cut-off value of 15.35, and the area under the curve (AUC) was 0.627. The GM assay detected antigen in sera obtained on day 1 (5/12), day 3 (9/12), day 5 (12/12), and day 7 (12/12) post-infection, and thus had a sensitivity of 79.2% and a specificity of 100%. The ROC of the GM assay indicated that the optimal Ct cut-off value was 1.40 (AUC, 0.919). The GM assay was more sensitive than the qRT-PCR assay in diagnosing invasive pulmonary aspergillosis in rats.

Development and Validation of Real-time PCR to Determine Branchiostegus japonicus and B. albus Species Based on Mitochondrial DNA (Real-time PCR 분석법을 이용한 옥돔과 옥두어의 종 판별법 개발)

  • Chung, In Young;Seo, Yong Bae;Yang, Ji-Young;Kim, Gun-Do
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1331-1339
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    • 2017
  • DNA barcoding is the identification of a species based on the DNA sequence of a fragment of the cytochrome C oxidase subunit I (COI) gene in the mitochondrial genome. It is widely applied to assist with the sustainable development of fishery-product resources and the protection of fish biodiversity. This study attempted to verify horse-head fish (Branchiostegus japonicus) and fake horse-head fish (Branchiostegus albus) species, which are commonly consumed in Korea. For the validation of the two species, a real-time PCR method was developed based on the species' mitochondrial DNA genome. Inter-species variations in mitochondrial DNA were observed in a bioinformatics analysis of the mitochondrial genomic DNA sequences of the two species. Some highly conserved regions and a few other regions were identified in the mitochondrial COI of the species. In order to test whether variations in the sequences were definitive, primers that targeted the varied regions of COI were designed and applied to amplify the DNA using the real-time PCR system. Threshold-cycle (Ct) range results confirmed that the Ct ranges of the real-time PCR were identical to the expected species of origin. Efficiency, specificity and cross-reactivity assays showed statistically significant differences between the average Ct of B. japonicus DNA ($21.85{\pm}3.599$) and the average Ct of B. albus DNA ($33.49{\pm}1.183$) for confirming B. japonicus. The assays also showed statistically significant differences between the average Ct of B. albus DNA ($22.49{\pm}0.908$) and the average Ct of B. japonicus DNA ($33.93{\pm}0.479$) for confirming B. albus. The methodology was validated by using ten commercial samples. The genomic DNA-based molecular technique that used the real-time PCR was a reliable method for the taxonomic classification of animal tissues.

The Study of Land Surface Change Detection Using Long-Term SPOT/VEGETATION (장기간 SPOT/VEGETATION 정규화 식생지수를 이용한 지면 변화 탐지 개선에 관한 연구)

  • Yeom, Jong-Min;Han, Kyung-Soo;Kim, In-Hwan
    • Journal of the Korean Association of Geographic Information Studies
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    • v.13 no.4
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    • pp.111-124
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    • 2010
  • To monitor the environment of land surface change is considered as an important research field since those parameters are related with land use, climate change, meteorological study, agriculture modulation, surface energy balance, and surface environment system. For the change detection, many different methods have been presented for distributing more detailed information with various tools from ground based measurement to satellite multi-spectral sensor. Recently, using high resolution satellite data is considered the most efficient way to monitor extensive land environmental system especially for higher spatial and temporal resolution. In this study, we use two different spatial resolution satellites; the one is SPOT/VEGETATION with 1 km spatial resolution to detect coarse resolution of the area change and determine objective threshold. The other is Landsat satellite having high resolution to figure out detailed land environmental change. According to their spatial resolution, they show different observation characteristics such as repeat cycle, and the global coverage. By correlating two kinds of satellites, we can detect land surface change from mid resolution to high resolution. The K-mean clustering algorithm is applied to detect changed area with two different temporal images. When using solar spectral band, there are complicate surface reflectance scattering characteristics which make surface change detection difficult. That effect would be leading serious problems when interpreting surface characteristics. For example, in spite of constant their own surface reflectance value, it could be changed according to solar, and sensor relative observation location. To reduce those affects, in this study, long-term Normalized Difference Vegetation Index (NDVI) with solar spectral channels performed for atmospheric and bi-directional correction from SPOT/VEGETATION data are utilized to offer objective threshold value for detecting land surface change, since that NDVI has less sensitivity for solar geometry than solar channel. The surface change detection based on long-term NDVI shows improved results than when only using Landsat.

Correlation of Basal AMH & Ovarian Response in IVF Cycles; Predictive Value of AMH (과배란유도 시 혈중 AMH와 난소 반응성과의 상관관계; 예측 인자로서의 효용성)

  • Ahn, Young-Sun;Kim, Jin-Yeong;Cho, Yun-Jin;Kim, Min-Ji;Kim, Hye-Ok;Park, Chan-Woo;Song, In-Ok;Koong, Mi-Kyoung;Kang, Inn-Soo
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.4
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    • pp.309-317
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    • 2008
  • Objectives: The aim of this study was to evaluate the usefulness of Anti-mullerian hormone (AMH) as a predictive marker for ovarian response and cycle outcome in IVF cycles. Methods: From Jan., to Aug., 2007, 111 patients undergoing IVF/ICSI stimulated by short or antagonist protocol were selected. On cycle day 3, basal serum AMH level and FSH level were measured. The correlation between basal serum AMH or FSH, and COH outcome was analyzed and IVF outcome was compared according to the AMH levels. To determine the threshold value of AMH for poor- and hyper-response, ROC curve was analyzed. Results: Serum AMH showed higher correlation coefficient (r=0.792, p<0.001) with the number of retrieved mature oocyte than serum FSH (r=-0.477, p<0.001). According to ovarian response, FSH and AMH leves showed significant differences among poor, normal, and hyperresponder. For predicting poor (${\leq}2$ oocytes) and hyperresponse (${\geq}17$ oocyets), AMH cut-off values were 0.5 ng/ml (the sensitivity 88.9% and the specificity 89.5%) and 2.5 ng/ml (sensitivity 85.7%, specificity 87.0%), respectively. According to the AMH level, patients were divided into 3 groups: low (${\leq}0.60\;ng/ml$), normal ($0.60{\sim}2.60\;ng/ml$), and high AMH (${\geq}2.60\;ng/ml$). The number of retrieved mature oocytes was significantly higher ($2.7{\pm}2.2$, $8.1{\pm}4.8$, $16.5{\pm}5.7$) and total gonadotropin dose was lower ($3530.5{\pm}1251.0$, $2957.1{\pm}1057.6$, and $2219.2{\pm}751.9\;IU$) in high AMH group (p<0.001). There was no significant difference in fertilization rates and pregnancy rates (23.8%, 34.0%, 37.5%) among the groups. Conclusions: Basal serum AMH level correlated better with the number of retrieved mature oocytes than FSH level, suggesting its usefulness for predicting ovarian response. However, IVF outcome was not significantly different according to the AMH levels. Serum AMH level presented good cut-off value for poor- or hyper-responders, therefore it could be useful in prediction of cycle cancellation, gonadotropin dose, and OHSS risk in IVF cycles.

Viral Load Dynamics After Symptomatic COVID-19 in Children With Underlying Malignancies During the Omicron Wave

  • Ye Ji Kim;Hyun Mi Kang;In Young Yoo;Jae Won Yoo;Seong Koo Kim;Jae Wook Lee;Dong Gun Lee;Nack-Gyun Chung;Yeon-Joon Park;Dae Chul Jeong;Bin Cho
    • Pediatric Infection and Vaccine
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    • v.30 no.2
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    • pp.73-83
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    • 2023
  • Purpose: This study aimed to investigate the viral load dynamics in children with underlying malignancies diagnosed with symptomatic coronavirus disease 2019 (COVID-19). Methods: This was a retrospective longitudinal cohort study of patients <19 years old with underlying hemato-oncologic malignancies that were diagnosed with their first symptomatic severe acute respiratory syndrome coronavirus 2 polymerase chain reaction (PCR)-confirmed COVID-19 infection during March 1 to August 30, 2022. Review of electronic medical records and telephone surveys were undertaken to assess the clinical presentations and transmission route of the patients. Thresholds of negligible likelihood of infectious virus was defined as E gene reverse transcription (RT)-PCR cycle threshold (Ct) value ≥25. Results: During the 6-month study period, a total of 43 children with 44 episodes of COVID-19 were included. Of the 44 episodes, the median age of the patients included was 8 years old (interquartile range [IQR], 4.9-10.5), and the most common underlying disease was acute lymphoid leukemia (n=30, 68.2%), followed by patients post-hematopoietic stem cell transplantation (n=8, 18.2%). Majority of the patients had mild COVID-19 (n=32, 72.7%), and three patients (7.0%) had severe/critical COVID-19. Furthermore, 2.3% (n=1) died of COVID-19 associated acute respiratory distress syndrome. The largest percentage of the patients showed E gene RT-PCR Ct value ≥25 between 15-21 days (n=13, 39.4%), followed by 22-28 days (n=10, 30.3%). In 15.2% (n=5), E gene RT-PCR Ct value remained <25 beyond 28 days after initial positive PCR. Refractory malignancy status (β, 67.0; 95% confidence interval, 7.0-17.0; P=0.030) was significantly associated with prolonged duration of E gene RT-PCR <25. A patient with prolonged duration of E gene RT-PCR Ct value <25 was suspected to have infectivity shown by the transmission of the virus to his mother at day 86 after his initial positive test. Conclusions: Children that acquire symptomatic COVID-19 during refractory malignancy state are at a high risk for prolonged shedding warranting PCR-based transmission precautions in this cohort of patients.