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Development of reverse transcription loop-mediated isothermal amplification assays for point-of-care testing of avian influenza virus subtype H5 and H9

  • Zhang, Songzi;Shin, Juyoun;Shin, Sun;Chung, Yeun-Jun
    • Genomics & Informatics
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    • v.18 no.4
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    • pp.40.1-40.8
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    • 2020
  • Avian influenza (AIV) outbreaks can induce fatal human pulmonary infections in addition to economic losses to the poultry industry. In this study, we aimed to develop a rapid and sensitive point-of-care AIV test using loop-mediated isothermal amplification (LAMP) technology. We designed three sets of reverse transcription LAMP (RT-LAMP) primers targeting the matrix (M) and hemagglutinin (HA) genes of the H5 and H9 subtypes. RT-LAMP targeting the universal M gene was designed to screen for the presence of AIV and RT-LAMP assays targeting H5-HA and H9-HA were designed to discriminate between the H5 and H9 subtypes. All three RT-LAMP assays showed specific amplification results without nonspecific reactions. In terms of sensitivity, the detection limits of our RT-LAMP assays were 100 to 1,000 RNA copies per reaction, which were 10 times more sensitive than the detection limits of the reference reverse-transcription polymerase chain reaction (RT-PCR) (1,000 to 10,000 RNA copies per reaction). The reaction time of our RT-LAMP assays was less than 30 min, which was approximately four times quicker than that of conventional RT-PCR. Altogether, these assays successfully detected the existence of AIV and discriminated between the H5 or H9 subtypes with higher sensitivity and less time than the conventional RT-PCR assay.

A deep learning-based approach for feeding behavior recognition of weanling pigs

  • Kim, MinJu;Choi, YoHan;Lee, Jeong-nam;Sa, SooJin;Cho, Hyun-chong
    • Journal of Animal Science and Technology
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    • v.63 no.6
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    • pp.1453-1463
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    • 2021
  • Feeding is the most important behavior that represents the health and welfare of weanling pigs. The early detection of feed refusal is crucial for the control of disease in the initial stages and the detection of empty feeders for adding feed in a timely manner. This paper proposes a real-time technique for the detection and recognition of small pigs using a deep-leaning-based method. The proposed model focuses on detecting pigs on a feeder in a feeding position. Conventional methods detect pigs and then classify them into different behavior gestures. In contrast, in the proposed method, these two tasks are combined into a single process to detect only feeding behavior to increase the speed of detection. Considering the significant differences between pig behaviors at different sizes, adaptive adjustments are introduced into a you-only-look-once (YOLO) model, including an angle optimization strategy between the head and body for detecting a head in a feeder. According to experimental results, this method can detect the feeding behavior of pigs and screen non-feeding positions with 95.66%, 94.22%, and 96.56% average precision (AP) at an intersection over union (IoU) threshold of 0.5 for YOLOv3, YOLOv4, and an additional layer and with the proposed activation function, respectively. Drinking behavior was detected with 86.86%, 89.16%, and 86.41% AP at a 0.5 IoU threshold for YOLOv3, YOLOv4, and the proposed activation function, respectively. In terms of detection and classification, the results of our study demonstrate that the proposed method yields higher precision and recall compared to conventional methods.

Serum proteomics analysis of feline mammary carcinoma based on label-free and PRM techniques

  • Zheng, Jia-San;Wei, Ren-Yue;Wang, Zheng;Zhu, Ting-Ting;Ruan, Hong-Ri;Wei, Xue;Hou, Kai-Wen;Wu, Rui
    • Journal of Veterinary Science
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    • v.21 no.3
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    • pp.45.1-45.15
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    • 2020
  • Background: Feline mammary carcinoma is the third most common cancer that affects female cats. Objectives: The purpose of this study was to screen differential serum proteins in feline and clarify the relationship between them and the occurrence of feline mammary carcinoma. Methods: Chinese pastoral cats were used as experimental animals. Six serum samples from cats with mammary carcinoma (group T) and six serum samples from healthy cats (group C) were selected. Differential protein analysis was performed using a Label-free technique, while parallel reaction monitoring (PRM) was performed to verify the screened differential proteins. Results: A total of 82 differential proteins were detected between group T and group C, of which 55 proteins were down regulated and 27 proteins were up regulated. Apolipoprotein A-I, Apolipoprotein A-II (ApoA-II), Apolipoprotein B (ApoB), Apolipoprotein C-III (ApoC-III), coagulation factor V, coagulation factor X, C1q, albumen (ALB) were all associated with the occurrence of feline mammary carcinoma. Differential proteins were involved in a total of 40 signaling pathways, among which the metabolic pathways associated with feline mammary carcinoma were the complement and coagulation cascade and cholesterol metabolism. According to the Label-free results, ApoB, ApoC-III, ApoA-II, FN1, an uncharacterized protein, and ALB were selected for PRM target verification. The results were consistent with the trend of the label-free. Conclusions: This experimen is the first to confirm ApoA-II and ApoB maybe new feline mammary carcinoma biomarkers and to analyze their mechanisms in the development of such carcinoma in feline.

Effects of a Mixture of Cynanchi Wilfordii Radix and Humuli Lupuli Flos Extract on Estrogenic Activities and Anti-Osteoclastogenesis (백수오(白首烏)와 비주화(啤酒花) 복합물의 에스트로겐 활성과 파골세포 분화 억제효과)

  • Park, Dongjun;Lee, Hong Gu;Min, Kyoungin;Park, Hyoungkook;Jin, Mu Hyun;Cho, Ho Song
    • The Korea Journal of Herbology
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    • v.37 no.5
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    • pp.1-8
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    • 2022
  • Objectives : This study aimed to investigate the synergistic effect of combining Cynanchi Wilfordii Radix extract with Humuli Lupuli Flos extract on estrogenic and anti-osteoclastogenic activity. Methods : Estrogenic effect of a mixture of Cynanchi Wilfordii Radix extract and Humuli Lupuli Flos extract (CWHL), Cynanchi Wilfordii Radix extract, Humuli Lupuli Flos extract, caudatin (an active ingredient of Cynanchi wilfordii Radix extract) and 8-prenylnaringenin (an active ingredient of Humuli Lupuli Flos extract) were examined by proliferation E-screen assay and expression of estrogen inducible gene, pS2 via Real Time-PCR (RT-PCR) in MCF-7 estrogen responsive cells. And their estrogenic activities were investigated how to modulate Estrogen receptor 𝛽 by binding affinity assay. Inhibitory effect of CWHL, Cynanchi Wilfordii Radix extract, Humuli Lupuli Flos extract, caudatin and 8-prenylnaringenin on RANKL-induced osteoclast differentiation were tested by TRAP (Tartrate-resistant acid phosphatase) staining in osteoclastogenic RAW 264.7 cells. Results : CWHL, Humuli Lupuli Flos extract and 8-prenylnaringenin accelerated the proliferation of MCF-7 and the expression of pS2 in MCF-7. CWHL, Cynanchi Wilfordii Radix extract, Humuli Lupuli Flos extract, caudatin and 8-prenylnaringenin bind to estrogen receptor 𝛽. CWHL, Cynanchi Wilfordii Radix extract, Humuli Lupuli Flos extract, caudatin and 8-prenylnaringenin inhibited RANKL-induced osteoclastogenesis in osteoclastogenic RAW 264.7. CWHL is more effective for all markers than Cynanchi Wilfordii Radix extract or Humuli Lupuli Flos extract alone. Conclusions : CWHL may a potential therapeutic agent for menopause and osteoporosis as a natural food resource. CWHL as a natural food source has therapeutic potential in cases of menopause and osteoporosis.

Genetic Studies on Heading-to-Ripening Period and Its Relationship to Yield Components in Barley I. Studies on maturity criteria in barley (대맥의 등숙일수와 수량구성요소와의 관계에 대한 유전연구 제I보 대맥의 생리적 성숙기 기준 설정)

  • Chun; J.U.;Lee, E.S.;Lee, H.S.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.27 no.1
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    • pp.49-54
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    • 1982
  • Experiments were carried out to establish physiological maturity determination criteria with reference to visibly easy measurement in barley in 1980 at Suweon. Thirty-three cultivars and lines from 500 crossing blocks were classified into 4 heading groups, and 5 variables; moisture content, discoloration of awn, exsertion, lemma and flag leaf were measured. There were highly negative correlations between heading date and ripening periods (r=-0.656$^{**}$ ), so early heading types had longer ripening periods. Comparing with the variables used for maturity determination, moisture content and discoloration of lemma were most sensitive to development of grain-filling. Those two variables, alone or in combination could be used to screen many genotypes of barley for physiological maturity. In determination of maturity with reference to visibly easy measurement, color of lemma changed stably and was the most useful way and discolor of flag leaf increased the accuracy of determination. The color of lemma at this time was Grayish yellow, and the mean moisture content was about 33 percent in 33 barley cultivars and lines.

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Selective cytotoxicity of a novel mitochondrial complex I inhibitor, YK-135, against EMT-subtype gastric cancer cell lines due to impaired glycolytic capacity

  • Yeojin, Sung;Seungbin, Cha;Sang Bum, Kim;Hakhyun, Kim;Seonghwi, Choi;Sejin, Oh;Minseo, Kim;Yunji, Lee;Gino, Kwon;Jooyoung, Lee;Joo-Youn, Lee;Gyoonhee, Han;Hyun Seok, Kim
    • BMB Reports
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    • v.55 no.12
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    • pp.645-650
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    • 2022
  • Epithelial-to-mesenchymal transition (EMT)-subtype gastric cancers have the worst prognosis due to their higher recurrence rate, higher probability of developing metastases and higher chemo-resistance compared to those of other molecular subtypes. Pharmacologically actionable somatic mutations are rarely found in EMT-subtype gastric cancers, limiting the utility of targeted therapies. Here, we conducted a high-throughput chemical screen using 37 gastric cancer cell lines and 48,467 synthetic small-molecule compounds. We identified YK-135, a small-molecule compound that showed higher cytotoxicity toward EMT-subtype gastric cancer cell lines than toward non-EMT-subtype gastric cancer cell lines. YK-135 exerts its cytotoxic effects by inhibiting mitochondrial complex I activity and inducing AMP-activated protein kinase (AMPK)-mediated apoptosis. We found that the lower glycolytic capacity of the EMT-subtype gastric cancer cells confers synthetic lethality to the inhibition of mitochondrial complex I, possibly by failing to maintain energy homeostasis. Other well-known mitochondrial complex I inhibitors (e.g., rotenone and phenformin) mimic the efficacy of YK-135, supporting our results. These findings highlight mitochondrial complex I inhibitors as promising therapeutic agents for EMT-subtype gastric cancers and YK-135 as a novel chemical scaffold for further drug development.

Pseudoaneurysm formation or dural arteriovenous fistula formation at the middle meningeal artery following revascularization surgery in Moyamoya disease

  • Dongok Seo;Byul Hee Yoon;Joonho Byun;Wonhyoung Park;Jung Cheol Park;Jae Sung Ahn
    • Journal of Cerebrovascular and Endovascular Neurosurgery
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    • v.25 no.1
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    • pp.87-92
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    • 2023
  • Moyamoya disease (MMD) is a rare progressive steno-occlusive cerebrovascular disorder. Currently, revascularization surgery is used as optimal treatment to overcome MMD. However, revascularization for MMD has reported several complications. Also, iatrogenic complications such as pseudoaneurysms formation or dural arteriovenous fistulas (dAVFs) formation-has been identified in rare cases after the surgical intervention for revascularizations. We describe two cases. In first case, the patency of the anastomosis site was good and saccular type pseudoaneurysm formation was found at parietal branch of posterior middle meningeal artery (MMA) in transfemoral cerebral angiography (TFCA) performed on the twelfth day after surgery. We decided to treat pseudoaneurysm by endovascular embolization the next day, but the patient was shown unconsciousness and anisocoria during sleep at that day. Computed tomography showed massive subdural hemorrhage at the ipsilateral side, thus we performed decompressive craniectomy and hematoma evacuation. In second case, the patency of the anastomosis site was good and dAVF formation at right MMA was found in TFCA performed on the sixth day after surgery. We performed endovascular obliteration of the arteriovenous fistula under local anesthesia. Pseudoaneurysm formation or dAVF formation after revascularization surgery is an exceptional case. If patients have such complications, practioner should carefully screen the patients by implementing digital subtraction angiogram to identify anatomic features; as well as consider immediate treatment in any way, including embolization or other surgery

The effect of job insecurity, employment type and monthly income on depressive symptom: analysis of Korean Longitudinal Study on Aging data

  • Myeong-Hun Lim;Jong-Uk Won;Won-Tae Lee;Min-Seok Kim;Seong-Uk Baek;Jin-Ha Yoon
    • Annals of Occupational and Environmental Medicine
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    • v.34
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    • pp.24.1-24.13
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    • 2022
  • Background: In modern society, depression is serious issue that causes socioeconomic and family burden. To decrease the incidence of depression, risk factors should be identified and managed. Among many risk factors for depression, this study examined socioeconomic risk factors for depression. Methods: We utilized first (2006), second (2008), and third (2010)-wave data from the Korean Longitudinal Study of Aging (KLoSA). Depressive symptom was measured with the 10-item Center for Epidemiological Studies Depression Scale, Short Form (CES-D-10) in the survey in 2008 and 2010. Three risk factors including job security, employment type and monthly income were measured in the survey in 2006. The association between risk factors and depressive symptom was analyzed by Cox proportional-hazard model. Results: We analyzed data from 1,105 workers and hazard ratios (HRs) for 3 risk factors were significant entirely. In addition, regular worker with high income group is the most vulnerable group of poor job insecurity on depression among male workers (HR: 1.73; 95% confidence interval [CI]: 1.07-2.81). Finally, HRs for 7 groups who had at least 1 risk factor had higher HRs compared to groups who had no risk factors after stratifying 3 risk factors. In the analysis, significantly vulnerable groups were total 5 groups and the group who had highest HR was temporary/daily workers with poor job security (HR: 2.51; 95% CI: 1.36-4.64). The results concerning women, regardless of job type, were non-significant. Conclusions: This study presented one or more risk factors among poor job security, low income, temporary/daily employment type increase hazard for depressive symptom in 2 or 4 years after the exposure. These results inform policy to screen for and protect against the risk of depression in vulnerable groups.

The heterotrimeric kinesin-2 family member KIF3A directly binds to disabled-1 (Dab1)

  • Myoung Hun Kim;Young Joo Jeong;Sang-Hwa Urm;Dae-Hyun Seog
    • BMB Reports
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    • v.57 no.10
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    • pp.447-452
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    • 2024
  • The heterotrimeric molecular motor kinesin-2 is involved in the microtubule-dependent transport of intracellular cargo. It consists of two distinct motor subunits (KIF3A, and KIF3B) and a non-motor subunit, kinesin-associated protein 3 (KAP3). The cargo-binding domain (CBD) at the carboxyl (C)-terminus of KIF3s plays an important role in the interaction with several different binding proteins. To identify the binding proteins for heterotrimeric kinesin-2, we performed a yeast two-hybrid screen and found a new interaction with Disables-1 (Dab1), the intracellular adaptor protein of reelin receptors. Dab1 bound to the CBD of KIF3A, but did not interact with the C-terminal domain of KIF3B, KIF5B, KIF17 or KAP3. The phosphotyrosine binding (PTB) domain-containing region of Dab1 is essential for the interaction with KIF3A. KIF3A interacted with GST-Dab1, and GST-CaMKIIα , but did not interact with GST-apolipoprotein E receptor 2 (ApoER2)-C or with GST alone. When co-expressed in HEK-293T cells, Dab1 co-precipitated with KIF3A, but not with KIF5B. Dab1 and KIF3A were co-localized in cultured cells. We also identified deduced cell surface expression of ApoER2 in KIF3A dominant-negative cells. These results suggest that the KIF3A plays a role in the intracellular trafficking of ApoER2 to the cell surface.

Screening of Extracts from Marine Green and Brown Algae in Jeju for Potential Marine Angiotensin-I Converting Enzyme (ACE) Inhibitory Activity (제주 자생 해양 녹조류와 갈조류 추출물로부터의 항고혈압 활성)

  • Cha, Seon-Heui;Ahn, Gin-Nae;Heo, Soo-Jin;Kim, Kil-Nam;Lee, Ki-Wan;Song, Choon-Bok;K.Cho, So-Mi;Jeon, You-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.3
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    • pp.307-314
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    • 2006
  • This study was conducted to screen in vitro angiotensin converting enzyme (ACE) inhibitory activities of methanol (MeOH) and aqueous extracts which were prepared by four different extractions-80% methanol extracts(ME) at $20^{\circ}C\;and\;70^{\circ}C$, respectively and aqueous extracts (AE) at both temperatures with the residue of the MEs-of ten marine green algae and nineteen brown algae collected along Jeju coast of Korea. Most marine brown algae extracts showed higher capacities than those of marine green algae in ACE inhibitory activity. Particularly, $70^{\circ}C$ MeOH extract (70ME) of Hizikia fusiforme showed the strongest inhibition activity (about 87%) among all the extracts. Also, 70 MEs of Enteromorpha linza, Ishige sinicola, Laminaria ochotensis, Petrospongium rugosum, Sagrassum horneri, Undaria pinnatifida and $20^{\circ}C$ MeOH extracts (20ME) of Myagropsis myagroides, Petrospongium rugosum, $20^{\circ}C$ aqueous extracts (20AE) of Codium contractum, Enteromorpha compressa, and $70^{\circ}C$ aqueous extracts (70AE) of Ecklonia cava, Petrospongium rugosum showed moderate ACE inhibitory activities more than 50% and the other extracts exhibited weak activities. On tile other hand, E. cava had the best ACE inhibitory activity among 70AEs. This indicates that 70AE of E. cava contains potential anti-ACE macromolecular. We tried to proteolytic digest 70AE of E. cava to induce production of anti-ACE peptides from E. cava 70AE. The enzymes used are five pretenses including Kojizyme, Flavourzyme, Neutrase, Alcalase, and Protamex, which are food grade-commercial enzymes from Novo Co. Flavourzyme-digest of E. cava 70AE showed the highest inhibitory activity about 90%. And the five different enzymatic digests of the E. cava 70AE ranged from 2.33 to 3.56 ${\mu}g/mL$, respectively in $IC_{50}$ values of anti-ACE activity.