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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.

Predictors and Prevalence of Alcohol and Cannabis Co-use Among Filipino Adolescents: Evidence From a School-based Student Health Survey

  • Yusuff Adebayo Adebisi;Don Eliseo Lucero-Prisno III;Jerico B. Ogaya;Victor C. Canezo Jr.;Roland A. Niez;Florante E. Delos Santos;Melchor M. Magramo;Ann Rosanie Yap-Tan;Francis Ann R. Sy;Omar Kasimieh
    • Journal of Preventive Medicine and Public Health
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    • v.57 no.3
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    • pp.288-297
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    • 2024
  • Objectives: This study explored the prevalence and predictors of alcohol and cannabis co-use among 9263 Filipino adolescents, using data from the 2019 Global School-based Student Health Survey (GSHS). Methods: We conducted a cross-sectional secondary analysis of the GSHS, targeting adolescents aged 13-17 years and excluding cases with incomplete data on alcohol and cannabis use. Our analysis employed the bivariate chi-square test of independence and multivariable logistic regression using Stata version 18 to identify significant predictors of co-use, with a p-value threshold set at 0.05. Results: The weighted prevalence of co-users was 4.2% (95% confidence interval [CI], 3.4 to 5.3). Significant predictors included male sex (adjusted odds ratio [aOR], 4.50; 95% CI, 3.31 to 6.10; p<0.001) and being in a lower academic year, specifically grade 7 (aOR, 4.08; 95% CI, 2.39 to 6.99; p<0.001) and grade 8 (aOR, 2.20; 95% CI, 1.30 to 3.72; p=0.003). Poor sleep quality was also a significant predictor (aOR, 1.77; 95% CI, 1.29 to 2.44; p<0.001), as was a history of attempted suicide (aOR, 5.31; 95% CI, 4.00 to 7.06; p<0.001). Physical inactivity was associated with lower odds of co-use (aOR, 0.45; 95% CI, 0.33 to 0.62; p<0.001). Additionally, non-attendance of physical education classes (aOR, 1.48; 95% CI, 1.06 to 2.05; p=0.021), infrequent unapproved parental checks (aOR, 1.37; 95% CI, 1.04 to 1.80; p=0.024), and lower parental awareness of free-time activities (aOR, 0.63; 95% CI, 0.45 to 0.87; p=0.005) were associated with higher odds of co-use. Factors not significantly linked to co-use included age group, being in grade 9, always feeling lonely, having no close friends, being bullied outside school, and whether a parent or guardian understood the adolescent's worries. Conclusions: The findings highlight the critical need for comprehensive interventions in the Philippines, addressing not only physical inactivity and parental monitoring but also focusing on sex, academic grade, participation in physical education classes, sleep quality, and suicide attempt history, to effectively reduce alcohol and cannabis co-use among adolescents.

Decay-Accelerating Factor Differentially Associates With Complement-Mediated Damage in Synovium After Meniscus Tear as Compared to Anterior Cruciate Ligament Injury

  • V. Michael Holers;Rachel M. Frank;Michael Zuscik;Carson Keeter;Robert I. Scheinman;Christopher Striebich;Dmitri Simberg;Michael R. Clay;Larry W. Moreland;Nirmal K. Banda
    • IMMUNE NETWORK
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    • v.24 no.2
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    • pp.17.1-17.16
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    • 2024
  • We have reported that anterior cruciate ligament (ACL) injury leads to the differential dysregulation of the complement system in the synovium as compared to meniscus tear (MT) and proposed this as a mechanism for a greater post-injury prevalence of post traumatic osteoarthritis (PTOA). To explore additional roles of complement proteins and regulators, we determined the presence of decay-accelerating factor (DAF), C5b, and membrane attack complexes (MACs, C5b-9) in discarded surgical synovial tissue (DSST) collected during arthroscopic ACL reconstructive surgery, MT-related meniscectomy, osteoarthritis (OA)-related knee replacement surgery and normal controls. Multiplexed immunohistochemistry was used to detect and quantify complement proteins. To explore the involvement of body mass index (BMI), after these 2 injuries, we examined correlations among DAF, C5b, MAC and BMI. Using these approaches, we found that synovial cells after ACL injury expressed a significantly lower level of DAF as compared to MT (p<0.049). In contrast, C5b staining synovial cells were significantly higher after ACL injury (p<0.0009) and in OA DSST (p<0.039) compared to MT. Interestingly, there were significantly positive correlations between DAF & C5b (r=0.75, p<0.018) and DAF & C5b (r=0.64 p<0.022) after ACL injury and MT, respectively. The data support that DAF, which should normally dampen C5b deposition due to its regulatory activities on C3/C5 convertases, does not appear to exhibit that function in inflamed synovia following either ACL injury or MT. Ineffective DAF regulation may be an additional mechanism by which relatively uncontrolled complement activation damages tissue in these injury states.

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.

Evaluation of the correlation between the muscle fat ratio of pork belly and pork shoulder butt using computed tomography scan

  • Sheena Kim;Jeongin Choi;Eun Sol Kim;Gi Beom Keum;Hyunok Doo;Jinok Kwak;Sumin Ryu;Yejin Choi;Sriniwas Pandey;Na Rae Lee;Juyoun Kang;Yujung Lee;Dongjun Kim;Kuk-Hwan Seol;Sun Moon Kang;In-Seon Bae;Soo-Hyun Cho;Hyo Jung Kwon;Samooel Jung;Youngwon Lee;Hyeun Bum Kim
    • Korean Journal of Agricultural Science
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    • v.50 no.4
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    • pp.809-815
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    • 2023
  • This study was conducted to find out the correlation between meat quality and muscle fat ratio in pork part meat (pork belly and shoulder butt) using CT (computed tomography) imaging technique. After 24 hours from slaughter, pork loin and belly were individually prepared from the left semiconductors of 26 pigs for CT measurement. The image obtained from CT scans was checked through the picture archiving and communications system (PACS). The volume of muscle and fat in the pork belly and shoulder butt of cross-sectional images taken by CT was estimated using Vitrea workstation version 7. This assemblage was further processed through Vitrea post-processing software to automatically calculate the volumes (Fig. 1). The volumes were measured in milliliters (mL). In addition to volume calculation, a three-dimensional reconstruction of the organ under consideration was generated. Pearson's correlation coefficient was analyzed to evaluate the relationship by region (pork belly, pork shoulder butt), and statistical processing was performed using GraphPad Prism 8. The muscle-fat ratios of pork belly taken by CT was 1 : 0.86, while that of pork shoulder butt was 1 : 0.37. As a result of CT analysis of the correlation coefficient between pork belly and shoulder butt compared to the muscle-fat ratio, the correlation coefficient was 0.5679 (R2 = 0.3295, p < 0.01). CT imaging provided very good estimates of muscle contents in cuts and in the whole carcass.

Evaluating the Properties and Commercializing Potential Of Rape Stalk-based Pellets Produced with a Pilot-scaled Flat-die Pellet Mill (파일럿 규모의 평다이 성형기로 제조한 유채대 펠릿의 연료적 특성 및 상용화 가능성 평가)

  • Sei Chang Oh;In Yang
    • Korean Chemical Engineering Research
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    • v.62 no.1
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    • pp.80-86
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    • 2024
  • This study was conducted to evaluate the potential of rape stalk (RAS) as a raw material for the production of solid bio-fuels. RAS was immersed in an aqueous solution with acetic acid concentration of 1 percent, The content of reducing sugars separated from the RAS was analyzed. Glucose showed the highest content followed by xylose, galactose, arabinose and mannose. The immersed and non-immersed RAS were used for producing pellets with a pilot-scaled flat-die pellet mill. Bulk density and calorific values of the pellets improved with the use of the immersed RAS and the addition of wood particles. The values exceeded the minimum requirements for the A-grade of non-woody pellets (≧600 kg/m3 & ≧ 14.5 MJ/kg) designated by the ISO. Ash content of the pellets reduced with the immersion of RAS and the value satisfied the A-grade level (≦6.0%) of the ISO standard. The durability of the immersed RAS-based pellets was much higher than that of non-immersed IRS-based pellets, and the values were increased with the addition of wood particles. However, the durability did not meet the acceptance level for the B-grade of non-woody pellets (≧96.0%) designated by the ISO. These results suggested that the addition of binders in the production of non-woody pellets using an RAS immersed in acetic acid-based aqueous solution is required for the commercialization of the pellets.

Stem Rot of Pearl Millet Prevalence, Symptomatology, Disease Cycle, Disease Rating Scale and Pathogen Characterization in Pearl Millet-Klebsiella Pathosystem

  • Vinod Kumar Malik;Pooja Sangwan;Manjeet Singh;Pavitra Kumari;Niharika Shoeran;Navjeet Ahalawat;Mukesh Kumar;Harsh Deep;Kamla Malik;Preety Verma;Pankaj Yadav;Sheetal Kumari;Aakash;Sambandh Dhal
    • The Plant Pathology Journal
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    • v.40 no.1
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    • pp.48-58
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    • 2024
  • The oldest and most extensively cultivated form of millet, known as pearl millet (Pennisetum glaucum (L.) R. Br. Syn. Pennisetum americanum (L.) Leeke), is raised over 312.00 lakh hectares in Asian and African countries. India is regarded as the significant hotspot for pearl millet diversity. In the Indian state of Haryana, where pearl millet is grown, a new and catastrophic bacterial disease known as stem rot of pearl millet spurred by the bacterium Klebsiella aerogenes (formerly Enterobacter) was first observed during fall 2018. The disease appears in form of small to long streaks on leaves, lesions on stem, and slimy rot appearance of stem. The associated bacterium showed close resemblance to Klebsiella aerogenes that was confirmed by a molecular evaluation based on 16S rDNA and gyrA gene nucleotide sequences. The isolates were also identified to be Klebsiella aerogenes based on biochemical assays, where Klebsiella isolates differed in D-trehalose and succinate alkalisation tests. During fall 2021-2023, the disease has spread all the pearl millet-growing districts of the state, extending up to 70% disease incidence in the affected fields. The disease is causing considering grain as well as fodder losses. The proposed scale, consisting of six levels (0-5), is developed where scores 0, 1, 2, 3, 4, and 5 have been categorized as highly resistant, resistant, moderately resistant, moderately susceptible, susceptible, and highly susceptible disease reaction, respectively. The disease cycle, survival of pathogen, and possible losses have also been studied to understand other features of the disease.

Quality Improvement of Crude Glycerol from Biodiesel Production Using Activated Carbon Derived from Krabok (Irvingia malayana) Seed Shells

  • Wuttichai Roschat;Sarunya Donrussamee;Phatcharanan Smanmit;Samlit Jikjak;Tappagorn Leelatam;Sunti Phewphong;Krittiyanee Namwongsa;Preecha Moonsin;Vinich Promarak
    • Korean Journal of Materials Research
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    • v.34 no.1
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    • pp.1-11
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    • 2024
  • This research investigated the preparation of activated carbon derived from Krabok (Irvingia malayana) seed shells to improve the quality of crude glycerol obtained during biodiesel production. The activated carbon was prepared using a dry chemical activation method with NaOH, utilizing an innovative biomass incinerator. The results revealed that the resulting KC/AC-two-step exhibited favorable physicochemical adsorption properties, with a high surface area of 758.72 m2/g and an iodine number of 611.10 mg/g. These values meet the criteria of the industrial product standard for activated carbon No. TIS 900-2004, as specified by the Ministry of Industry in Thailand. Additionally, the adsorption efficiency for methylene blue reached an impressive 99.35 %. This developed activated carbon was then used to improve the quality of crude glycerol obtained from biodiesel production. The experimental results showed that the KC/AC-two-step increased the purity of crude glycerol to 73.61 %. In comparison, commercially available activated carbon (C/AC) resulted in a higher crude glycerol purity of 81.19 %, as analyzed by the GC technique. Additionally, the metal content (Zn, Cu, Fe, Pb, Cd, and Na) in purified glycerol using KC/AC-two-step was below the standards for heavy metals permitted in food and cosmeceuticals by the Food and Drug Administration of Thailand and the European Committee for Food Contact Materials and Articles. As a result, it can be inferred that Krabok seed shells have favorable properties for producing activated carbon suitable as an adsorbent to enhance crude glycerol purity. Furthermore, the improved crude glycerol from this research has potential for various industrial applications.

Shipboard Verification Test of Onboard Carbon Dioxide Capture System (OCCS) Using Sodium Hydroxide(NaOH) Solution (가성소다(NaOH) 용액을 이용한 선상 이산화탄소 포집 장치의 선박 검증시험)

  • Gwang Hyun Lee;Hyung Ju Roh;Min woo Lee;Won Kyeong Son;Jae Yeoul Jeong;Tae-Hong Kim;Byung-Tak NAM;Jae-Ik Kim
    • Journal of the Society of Naval Architects of Korea
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    • v.61 no.1
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    • pp.51-60
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    • 2024
  • Hi Air Korea and Hanwha ocean are currently developing an Onboard Carbon dioxide Capture System (OCCS) to absorb CO2 emitted from ship's engine using a sodium hydroxide(NaOH) solution, and converting the resulting salt into a solid form through a chemical reaction with calcium oxide (CaO). The system process involves the following steps; 1)The reaction of CO2 gas absorption in water, 2)The reaction between carbonic acid (H2CO3) and NaOH solution to produce carbonate or bicarbonate, and 3)The reaction between carbonate or bicarbonate and CaO to form calcium carbonate (CaCO3). And ultimately, the solid material, CaCO3, is separated and discharged using a separator. The OCCS has been installed on an ship and the test results have confirmed significant reduction effects of CO2 in the ship's exhaust gas. A portion of the exhaust gas emitted from the engine was transferred to the OCCS using a blower. The flow rate of the transferred gas ranged from 800 to 1384 m3/hr, and the CO2 concentration in the exhaust gas was 5.1 vol% for VLSFO, 3.7 vol% for LNG and a 12 wt% NaOH solution was used. The results showed a CO2 capture efficiency of approximately 42.5 to 64.1 vol% and the CO2 capture rate approximately 48.4 to 52.2kg/hr. Additionally, to assess the impact of the discharged CaCO3on the marine ecosystem, we conducted "marine ecotoxicity test" and performed Computational Fluid Dynamics (CFD) analysis to evaluate the dispersion and dilution of the discharged effluent.

Inhibitory Effects of Ssanghwa-tang on Lung Injury and Muscle Loss in a Cigarette Smoke Extract and Lipopolysaccharide-induced Chronic Obstructive Pulmonary Disease Mouse Model (표준담배추출물과 Lipopolysaccharide로 유발한 만성폐쇄성폐질환 동물모델에서 쌍화탕의 폐손상 및 근감소 억제 효과)

  • Jin-kwan Choi;Won-kyung Yang;Su-won Lee;Seong-cheon Woo;Seung-hyung Kim;Yang-chun Park
    • The Journal of Internal Korean Medicine
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    • v.45 no.1
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    • pp.11-30
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    • 2024
  • Objectives: This study evaluated the effects of Ssanghwa-tang (SHT) on lung injury and muscle loss in a COPD mouse model. Methods: C57BL/6 mice were challenged with cigarette smoke extract and lipopolysaccharide, and then treated with two concentrations of SHT (250 and 500 mg/kg). After sacrifice, the bronchoalveolar lavage fluid (BALF) or lung tissue was analyzed by cytospin, ELISA, real-time PCR, flow cytometry analysis, and H&E and Masson's trichrome staining. The grip strength of COPD mice was measured using a grip strength meter. The running time of COPD mice was measured by a treadmill test. Muscle tissue of the quadriceps was stained with H&E and Masson's trichrome staining. Results: SHT significantly inhibited the increase in neutrophil numbers in BALF and significantly decreased immune cell activity in BALF and lung tissue. It also significantly inhibited the increase in TNF-α, IL-17, and MIP2 in BALF. Real-time PCR analysis revealed that the mRNA expression of TNF-α, IL-17, MIP2, and TRPV1 in lung tissue showed a significant decrease compared with the control group. Lung tissue damage was significantly reduced in the histological analysis. The grip strength and running time of the COPD mice showed a significant decrease compared with the control group. In histological staining, SHT was found to reduce the damage to muscle tissue. Conclusions: This study indicates that SHT can be used as a therapeutic agent for COPD patients by inhibiting lung injury and muscle loss.