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Comparative pluripotent characteristics of porcine induced pluripotent stem cells generated using different viral transduction systems

  • Sang-Ki Baek;In-Won Lee;Yeon-Ji Lee;Bo-Gyeong Seo;Jung-Woo Choi;Tae-Suk Kim;Cheol Hwangbo;Joon-Hee Lee
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.4
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    • pp.275-290
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    • 2023
  • Background: Porcine pluripotent stem cells (pPSCs) would provide enormous potential for agriculture and biomedicine. However, authentic pPSCs have not established yet because standards for pPSCs-specific markers and culture conditions are not clear. Therefore, the present study reports comparative pluripotency characteristics in porcine induced pluripotent stem cells (piPSCs) derived from different viral transduction and reprogramming factors [Lenti-iPSCs (OSKM), Lenti-iPSCs (OSKMNL) and Sev-iPSCs (OSKM)]. Methods: Porcine fibroblasts were induced into Lenti-iPSCs (OSKM) and Lenti-iPSCs (OSKMNL) by using Lentiviral vector and Sev-iPSCs (OSKM) by using Sendaiviral vector. Expressions of endogenous or exogenous pluripotency-associated genes, surface marker and in vitro differentiation in between Lenti-piPSCs (OSKM), Lenti-iPSCs (OSKMNL) and Sev-piPSCs (OSKM) were compared. Results: Colonial morphology of Lenti-iPSCs (OSKMNL) closely resembles the naïve mouse embryonic stem cells colony for culture, whereas Sev-iPSCs (OSKM) colony is similar to the primed hESCs. Also, the activity of AP shows a distinct different in piPSCs (AP-positive (+) Lenti-iPSCs (OSKMNL) and Sev-iPSCs (OSKM), but AP-negative (-) Lenti-iPSCs (OSKM)). mRNAs expression of several marker genes (OCT-3/4, NANOG and SOX2) for pluripotency was increased in Lenti-iPSCs (OSKMNL) and Sev-iPSCs (OSKM), but Sev-iPSCs (OSKM). Interestingly, SSEA-1 of surface markers was expressed only in Sev-iPSCs (OSKM), whereas SSEA-4, Tra-1-60 and Tra-1-81 were positively expressed in Lenti-iPSCs (OSKMNL). Exogenous reprogramming factors continuously expressed in Lenti-iPSCs (OSKMNL) for passage 20, whereas Sev-iPSCs (OSKM) did not express any exogenous transcription factors. Finally, only Lenti-iPSCs (OSKMNL) express the three germ layers and primordial germ cells markers in aggregated EBs. Conclusions: These results indicate that the viral transduction system of reprograming factors into porcine differentiated cells display different pluripotency characteristics in piPSCs.

Effectiveness of cephalosporins in hydrolysis and inhibition of Staphylococcus aureus and Escherichia coli biofilms

  • Jawaria Aslam;Hafiz Muhammad Ali;Shujaat Hussain;Muhammad Zishan Ahmad;Abu Baker Siddique;Muhammad Shahid;Mirza Imran Shahzad;Hina Fatima;Sarah Tariq;Fatima Sadiq;Maria Aslam;Umar Farooq;Saadiya Zia;Rawa Saad Aljaluod;Khaloud Mohammed Alarjani
    • Journal of Veterinary Science
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    • v.25 no.3
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    • pp.47.1-47.12
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    • 2024
  • Importance: Staphylococcus aureus and Escherichia coli contribute to global health challenges by forming biofilms, a key virulence element implicated in the pathogenesis of several infections. Objective: The study examined the efficacy of various generations of cephalosporins against biofilms developed by pathogenic S. aureus and E. coli. Methods: The development of biofilms by both bacteria was assessed using petri-plate and microplate methods. Biofilm hydrolysis and inhibition were tested using first to fourth generations of cephalosporins, and the effects were analyzed by crystal violet staining and phase contrast microscopy. Results: Both bacterial strains exhibited well-developed biofilms in petri-plate and microplate assays. Cefradine (first generation) showed 76.78% hydrolysis of S. aureus biofilm, while significant hydrolysis (59.86%) of E. coli biofilm was observed by cefipime (fourth generation). Similarly, cefuroxime, cefadroxil, cefepime, and cefradine caused 78.8%, 71.63%, 70.63%, and 70.51% inhibition of the S. aureus biofilms, respectively. In the case of E. coli, maximum biofilm inhibition (66.47%) was again shown by cefepime. All generations of cephalosporins were more effective against S. aureus than E. coli, which was confirmed by phase contrast microscopy. Conclusions and Relevance: Cephalosporins exhibit dual capabilities of hydrolyzing and inhibiting S. aureus and E. coli biofilms. First-generation cephalosporins exhibited the highest inhibitory activity against S. aureus, while the third and fourth generations significantly inhibited E. coli biofilms. This study highlights the importance of tailored antibiotic strategies based on the biofilm characteristics of specific bacterial strains.

SARS-CoV-2 mRNA Vaccine Elicits Sustained T Cell Responses Against the Omicron Variant in Adolescents

  • Sujin Choi;Sang-Hoon Kim;Mi Seon Han;Yoonsun Yoon;Yun-Kyung Kim;Hye-Kyung Cho;Ki Wook Yun;Seung Ha Song;Bin Ahn;Ye Kyung Kim;Sung Hwan Choi;Young June Choe;Heeji Lim;Eun Bee Choi;Kwangwook Kim;Seokhwan Hyeon;Hye Jung Lim;Byung-chul Kim;Yoo-kyoung Lee;Eun Hwa Choi;Eui-Cheol Shin;Hyunju Lee
    • IMMUNE NETWORK
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    • v.23 no.4
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    • pp.33.1-33.13
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    • 2023
  • Vaccination against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been acknowledged as an effective mean of preventing infection and hospitalization. However, the emergence of highly transmissible SARS-CoV-2 variants of concern (VOCs) has led to substantial increase in infections among children and adolescents. Vaccine-induced immunity and longevity have not been well defined in this population. Therefore, we aimed to analyze humoral and cellular immune responses against ancestral and SARS-CoV-2 variants after two shots of the BNT162b2 vaccine in healthy adolescents. Although vaccination induced a robust increase of spike-specific binding Abs and neutralizing Abs against the ancestral and SARS-CoV-2 variants, the neutralizing activity against the Omicron variant was significantly low. On the contrary, vaccine-induced memory CD4+ T cells exhibited substantial responses against both ancestral and Omicron spike proteins. Notably, CD4+ T cell responses against both ancestral and Omicron strains were preserved at 3 months after two shots of the BNT162b2 vaccine without waning. Polyfunctionality of vaccine-induced memory T cells was also preserved in response to Omicron spike protein. The present findings characterize the protective immunity of vaccination for adolescents in the era of continuous emergence of variants/subvariants.

The Historical Astronomic Observatory and Calendar of the Village of Graw, Northern Iraq

  • Rzger Abdulkarim ABDULA
    • Acta Via Serica
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    • v.8 no.2
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    • pp.25-52
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    • 2023
  • The astronomic observatory of Graw Village is located on Mount Dari Lolikan, facing the village. Graw is located in the foothills of Mount Ser-i-Rash, 25 km northeast of Erbil Governorate, Iraq. This study attempts to clarify the foundations of this observatory, its components, as well as the founder and the date of its establishment. The study made efforts to clarify the benefits of this calendar to local residents in their daily lives. The database for this study is based on direct observation of the observatory station. The observation included the recording date and position of sunset and the appearance of stars throughout the year. Observation and documentation for both sunset and stars were performed over several years due to weather conditions since observation was not possible on foggy and rainy days and nights. Each observation took five to ten minutes depending on the clarity of the sky. The observatory consists of a group of stone cones. Each cone was built by stones in a specific location after careful and long observation of the sunset. Efforts were made to observe the disappearance and reappearance of the stars based on the change in the position of the Earth in relation to the sun. Graw's calendar helped to recognize important times of the year, such as the winter and summer forties, which were very important, especially when snow covered the roads, transportation stopped, crops spoiled, and pets stayed in their barn. The most important features of the winter forties are the memories, experiences, and minds of the villagers' ancestors. The forties were associated with the arrival of cold and heavier rain throughout the year, which is consistent with modern science, as the angle at which the Earth rotates increases the number and activity of weather depressions that affect the study area during this period. This observatory has a close connection with the daily life of the villagers, especially in the past centuries. It helped the people of the area in their appointments to carry out their work in the field of agriculture. The observatory was also of great importance in the field of education in the past centuries, especially in traditional religious schools. It also appears from this research that the calendar has ancient roots, which extend back thousands of years, as evidenced by the Ezidis who follow an ancient religion whose roots extend back thousands of years and who fast during both the winter and summer forties annually, with the participation of people in various regions of the world. It is not known who made this astronomic observatory but most of the oral information that has been passed down to us by word of mouth agrees on both Mullah Abdullah Al-Kurdi and Mullah Omar. Likely, this astronomic observatory was built around the late 17th and early 18th centuries.

Indoleamine 2,3-Dioxygenase in Hematopoietic Stem Cell-Derived Cells Suppresses Rhinovirus-Induced Neutrophilic Airway Inflammation by Regulating Th1- and Th17-Type Responses

  • Ferdaus Mohd Altaf Hossain;Seong Ok Park;Hyo Jin Kim;Jun Cheol Eo;Jin Young Choi;Maryum Tanveer;Erdenebelig Uyangaa;Koanhoi Kim;Seong Kug Eo
    • IMMUNE NETWORK
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    • v.21 no.4
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    • pp.26.1-26.28
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    • 2021
  • Asthma exacerbations are a major cause of intractable morbidity, increases in health care costs, and a greater progressive loss of lung function. Asthma exacerbations are most commonly triggered by respiratory viral infections, particularly with human rhinovirus (hRV). Respiratory viral infections are believed to affect the expression of indoleamine 2,3-dioxygenase (IDO), a limiting enzyme in tryptophan catabolism, which is presumed to alter asthmatic airway inflammation. Here, we explored the detailed role of IDO in the progression of asthma exacerbations using a mouse model for asthma exacerbation caused by hRV infection. Our results reveal that IDO is required to prevent neutrophilic inflammation in the course of asthma exacerbation caused by an hRV infection, as corroborated by markedly enhanced Th17- and Th1-type neutrophilia in the airways of IDO-deficient mice. This neutrophilia was closely associated with disrupted expression of tight junctions and enhanced expression of inflammasome-related molecules and mucin-inducing genes. In addition, IDO ablation enhanced allergen-specific Th17- and Th1-biased CD4+ T-cell responses following hRV infection. The role of IDO in attenuating Th17- and Th1-type neutrophilic airway inflammation became more apparent in chronic asthma exacerbations after repeated allergen exposures and hRV infections. Furthermore, IDO enzymatic induction in leukocytes derived from the hematopoietic stem cell (HSC) lineage appeared to play a dominant role in attenuating Th17- and Th1-type neutrophilic inflammation in the airway following hRV infection. Therefore, IDO activity in HSC-derived leukocytes is required to regulate Th17- and Th1-type neutrophilic inflammation in the airway during asthma exacerbations caused by hRV infections.

Anti-thrombotic effect of artemisinin through regulation of cAMP production and Ca2+ mobilization in U46619-induced human platelets (U46619 유도의 사람 혈소판에서 cAMP 생성 및 Ca2+동원의 조절을 통한 Artemisinin의 항혈전 효과)

  • Chang-Eun Park;Dong-Ha Lee
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.402-407
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    • 2023
  • The regulation of platelet aggregation is crucial for maintaining normal hemostasis, but abnormal or excessive platelet aggregation can contribute to cardiovascular disorders such as stroke, atherosclerosis, and thrombosis. Therefore, identifying substances that can control or suppress platelet aggregation is a promising approach for the prevention and treatment of these conditions. Artemisinin, a compound derived from Artemisia or Scopolia plants, has shown potential in various areas such as anticancer and Alzheimer's disease research. However, the specific role and mechanisms by which artemisinin influences platelet activation and thrombus formation are not yet fully understood. This study investigated the effects of artemisinin on platelet activation and thrombus formation. As a result, cAMP production were increased significantly by artemisinin, as well as phosphorylated VASP and IP3R which are substrates to cAMP-dependent kinase by artemisinin in a significant manner. The Ca2+ normally mobilized from the dense tubular system was inhibited due to IP3R phosphorylation from artemisinin, and phosphorylated VASP by artemisinin aided in inhibiting platelet activity via αIIb/β3 platelet membrane inactivation and inhibiting fibrinogen binding. Finally, artemisinin inhibited thrombin-induced thrombus formation. Therefore, we suggest that artemisinin has importance with cardiovascular diseases stemming from the abnormal platelets activation and thrombus formation by acting as an effective prophylactic and therapeutic agent.

Discovery of a Novel Cellobiose Dehydrogenase from Cellulomonas palmilytica EW123 and Its Sugar Acids Production

  • Ake-kavitch Siriatcharanon;Sawannee Sutheeworapong;Sirilak Baramee;Rattiya Waeonukul;Patthra Pason;Akihiko Kosugi;Ayaka Uke;Khanok Ratanakhanokchai;Chakrit Tachaapaikoon
    • Journal of Microbiology and Biotechnology
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    • v.34 no.2
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    • pp.457-466
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    • 2024
  • Cellobiose dehydrogenases (CDHs) are a group of enzymes belonging to the hemoflavoenzyme group, which are mostly found in fungi. They play an important role in the production of acid sugar. In this research, CDH annotated from the actinobacterium Cellulomonas palmilytica EW123 (CpCDH) was cloned and characterized. The CpCDH exhibited a domain architecture resembling class-I CDH found in Basidiomycota. The cytochrome c and flavin-containing dehydrogenase domains in CpCDH showed an extra-long evolutionary distance compared to fungal CDH. The amino acid sequence of CpCDH revealed conservative catalytic amino acids and a distinct flavin adenine dinucleotide region specific to CDH, setting it apart from closely related sequences. The physicochemical properties of CpCDH displayed optimal pH conditions similar to those of CDHs but differed in terms of optimal temperature. The CpCDH displayed excellent enzymatic activity at low temperatures (below 30℃), unlike other CDHs. Moreover, CpCDH showed the highest substrate specificity for disaccharides such as cellobiose and lactose, which contain a glucose molecule at the non-reducing end. The catalytic efficiency of CpCDH for cellobiose and lactose were 2.05 × 105 and 9.06 × 104 (M-1 s-1), respectively. The result from the Fourier-transform infrared spectroscopy (FT-IR) spectra confirmed the presence of cellobionic and lactobionic acids as the oxidative products of CpCDH. This study establishes CpCDH as a novel and attractive bacterial CDH, representing the first report of its kind in the Cellulomonas genus.

Walking the Labyrinth in the Korean Church ('래버린스 걷기'에 대한 고찰과 그 활용 방안)

  • Sam Jun Ryu
    • Journal of Christian Education in Korea
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    • v.75
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    • pp.127-152
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    • 2023
  • Purpose of study: This study aims to explore 'labyrinth walking,' a lesser-known Christian spiritual practice within the Korean church. Additionally, it intends to propose ways for implementing labyrinth walking through literature reviews encompassing theory and practice. Research content and method: The article begins with its exploration by defining labyrinth walking from a historical perspective. It was a spiritual practice that existed in the Middle Ages, involving the traversal of a labyrinthine shape, typically a single path on the floor, often accompanied by prayer or contemplation. The contemporary spiritual practice of labyrinth walking was recovered and reinterpreted by Christian scholars and spiritual directors during the mid and late twentieth century. Additionally, within Christianity, labyrinth walking, rich with symbolism representing Jesus' suffering and salvation, baptism, pilgrimage, and purification, comprises five essential steps: preparing, walking with prayer or contemplation, remaining in the center, returning, and journaling and sharing. By merging pilgrimage, walking, prayer, and meditation, the practice of walking the labyrinth can enable people to have an opportunity for retreat, to encounter and be accompanied by God, to experience moments of serenity and healing, to reflect on themselves, to create life-transforming resolutions, and to build a sense of community. As a result, many Korean churches should acknowledge the value of and utilize labyrinth walking to nurture faith and spirituality. Conclusions and Suggestions: Accordingly, this paper suggests some practical ways to employ labyrinth walking as a form of prayer, part of a ritual or religious activity, an embodiment of pilgrimage, and a tranquil and nonverbal experience. Furthermore, the study recommends that Korean churches establish proper goals, consider essential principles for labyrinth design and installation, and follow specific guidelines while conducting labyrinth walks.

A Recent Insight into the Diagnosis and Screening of Patients with Fabry Disease (파브리병 환자의 진단과 선별검사의 최신지견)

  • Hye-Ran Yoon;Jihun Jo
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.24 no.1
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    • pp.17-25
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    • 2024
  • Fabry disease (FD) is an X-linked lysosomal storage disorder. It is caused by mutations in the α-galactosidase A gene, which results in deficient or absent activity of α-galactosidase A (α-Gal A). This leads to a progressive accumulation of globotriaosylceramide (Gb3) in various tissues. Manifestations of Fabry disease include serious and progressive impairment of renal and cardiac function. In addition, patients experience pain, gastrointestinal disturbance, transient ischaemic attacks, and strokes. Additional effects on the skin, eyes, ears, lungs, and bones are often seen. Reduced life expectancy and deadly consequences are being caused by cardiac involvement. Chaperone therapy or enzyme replacement therapy (ERT) are two disease-specific treatments for FD. Thus, early detection of FD is critical for decreasing morbidity and mortality. Globotriaosysphingosine (lyso-Gb3) for identifying atypical FD variants and highly sensitive troponin T (hsTNT) for detecting cardiac involvement are both significant diagnostic indicators. This review aimed to offer a basic resource for the early diagnosis and update on the diagnosis of having FD. We will also provide a general diagnostic algorithm and information on ERT and its accompanying treatments.

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A Discourse Analysis of Science Teachers' Scientific Modeling Activities: A Case from Earth Science Teacher Training (과학 모델링 활동에 나타난 교사의 담화 분석 -지구과학 교사 연수 사례-)

  • Heungjin Eom;Hyunjin Shim
    • Journal of The Korean Association For Science Education
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    • v.44 no.4
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    • pp.301-311
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    • 2024
  • We developed a small-group training program for in-service teachers focused on scientific modeling. We collected the discourses of the teachers who participated in the activity and analyzed them by type. The training program employed a collaborative approach in which a small group completed tasks and produced outputs based on the theme of 'galaxies and the Universe' to enable practical application in classes. Three in-service science teachers participated in the training program. Their discourses were recorded, transcribed, and classified into types based on individual turns and interaction units. The language expressions of the teachers reflected the unique characteristics of the teaching profession, with each participant having preferred language expression types, albeit with a generally low prevalence of specific language expression types across the participants. Differences in discourse characteristics related to the modeling theme, task presentation method, and model types, revealed that variations in the proportion of interaction unit types during the modeling design, build, and evaluation stages were primarily influenced by the teachers' familiarity with the modeling theme. While the task presentation method also influenced interaction types, model types had little impact on the distribution of interaction types. Considering these findings, training programs on modeling for in-service teachers should include a checklist to encourage sufficient interaction between participants as well as propose proper questions that can be effectively addressed through collaboration.