• Korean Journal of Poultry Science
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• v.44 no.1
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• pp.1-9
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• 2017

The objective of the present study was to determine the expression of genes associated with lipopolysaccharide (LPS)-induced stressor in two breeds of chickens: the Korean native chicken (KNC) and the White Leghorn chicken (WLH). Forty chickens per breed, aged 40 weeks, were randomly allotted to the control (CON, administered the saline vehicle) and LPS-injected stress groups. Samples were collected at 0 and 48 h post-LPS injection, and total RNA was extracted from the chicken livers for RNA microarray and quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In response to LPS, 1,044 and 1,193 genes were upregulated, and 1,000 and 1,072 genes were downregulated in the KNC and WLH, respectively, using a ${\geq}2$-fold cutoff change. A functional network analysis revealed that stress-related genes were downregulated in both KNC and WLH after LPS infection. The results obtained from the qRT-PCR analysis of mRNA expression of heat shock 90 (HSP90), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), activating transcription factor 4 (ATF4), sterol regulatory element-binding protein 1 (SREBP1), and X-box binding protein 1 (XBP1) were confirmed by the results of the microarray analysis. There was a significant difference in the expression of stress-associated genes between the control and LPS-injected KNC and WLH groups. The qRT-PCR analysis revealed that the stress-related $HSP90{\alpha}$ and HMGCR genes were downregulated in both LPS-injected KNC and WLH groups. However, the HSP70 and $HSP90{\beta}$ genes were upregulated only in the LPS-injected KNC group. The results suggest that the mRNA expression of stress-related genes is differentially affected by LPS stimulation, and some of the responses varied with the chicken breed. A better understanding of the LPS-induced infective stressors in chicken using the qRT-PCR and RNA microarray analyses may contribute to improving animal welfare and husbandry practices.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.9
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• pp.4517-4525
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• 2016

Background: Sperm DNA damage is underlying aetiology of poor implantation and pregnancy rates but also affects health of offspring and may also result in denovo mutations in germ line and post fertilization. This may result in complex diseases, polygenic disorders and childhood cancers. Childhood cancer like retinoblastoma (RB) is more prevalent in developing countries and the incidence of RB has increased more than three fold in India in the last decade. Recent studies have documented increased incidence of cancers in children born to fathers who consume alcohol in excess and tobacco or who were conceived by assisted conception. The aetiology of childhood cancer and increased disease burden in these children is lin ked to oxidative stress (OS) and oxidative DNA damage( ODD) in sperm of their fathers. Though several antioxidants are in use to combat oxidative stress, the effect of majority of these formulations on DNA is not known. Yoga and meditation cause significant decline in OS and ODD and aid in regulating OS levels such that reactive oxygen speues meditated signal transduction, gene expression and several other physiological functions are not disrupted. Thus, this study aimed to analyze sperm ODD as a possible etiological factor in childhood cancer and role of simple life style interventions like yoga and meditation in significantly decreasing seminal oxidative stress and oxidative DNA damage and thereby decreasing incidence of childhood cancers. Materials and Methods: A total of 131 fathers of children with RB (non-familial sporadic heritable) and 50 controls (fathers of healthy children) were recruited at a tertiary center in India. Sperm parameters as per WHO 2010 guidelines and reactive oxygen species (ROS), DNA fragmentation index (DFI), 8-hydroxy-2'-deoxy guanosine (8-OHdG) and telomere length were estimated at day 0, and after 3 and 6 months of intervention. We also examined the compliance with yoga and meditation practice and smoking status at each follow-up. Results: The seminal mean ROS levels (p<0.05), sperm DFI (p<0.001), 8-OHdG (p<0.01) levels were significantly higher in fathers of children with RB, as compared to controls and the relative mean telomere length in the sperm was shorter. Levels of ROS were significantly reduced in tobacco users (p<0.05) as well as in alcoholics (p<0.05) after intervention. DFI reduced significantly (p<0.05) after 6 months of yoga and meditation practice in all groups. The levels of oxidative DNA damage marker 8-OHdG were reduced significantly after 3 months (p<0.05) and 6 months (p<0.05) of practice. Conclusions: Our results suggest that OS and ODD DNA may contribute to the development of childhood cancer. This may be due to accumulation of oxidized mutagenic base 8OHdG, and elevated MDA levels which results in MDA dimers which are also mutagenic, aberrant methylation pattern, altered gene expression which affect cell proliferation and survival through activation of transcription factors. Increased mt DNA mutations and aberrant repair of mt and nuclear DNA due to highly truncatred DNA repair mechanisms all contribute to sperm genome hypermutability and persistant oxidative DNA damage. Oxidative stress is also associated with genome wide hypomethylation, telomere shortening and mitochondrial dysfunction leading to genome hypermutability and instability. To the best of our knowledge, this is the first study to report decline in OS and ODD and improvement in sperm DNA integrity following adoption of meditation and yoga based life style modification.This may reduce disease burden in next generation and reduce incidence of childhood cancers.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.7
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• pp.922-929
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• 2019

Objective: Mutations in low-density lipoprotein receptor (LDLR), which encodes a critical protein for cholesterol homeostasis and lipid metabolism in mammals, are involved in cardiometabolic diseases, such as familial hypercholesterolemia in pigs. Whereas microRNAs (miRNAs) can control LDLR regulation, their involvement in circulating cholesterol and lipid levels with respect to cardiometabolic diseases in pigs is unclear. We aimed to identify and analyze LDLR as a potential target gene of SSC-miR-20a. Methods: Bioinformatic analysis predicted that porcine LDLR is a target of SSC-miR-20a. Wild-type and mutant LDLR 3'-untranslated region (UTR) fragments were generated by polymerase chain reaction (PCR) and cloned into the pGL3-Control vector to construct pGL3 Control LDLR wild-3'-UTR and pGL3 Control LDLR mutant-3'-UTR recombinant plasmids, respectively. An miR-20a expression plasmid was constructed by inserting the porcine premiR-20a-coding sequence between the HindIII and BamHI sites in pMR-mCherry, and constructs were confirmed by sequencing. HEK293T cells were co-transfected with the miR-20a expression or pMR-mCherry control plasmids and constructs harboring the corresponding 3'-UTR, and relative luciferase activity was determined. The relative expression levels of miR-20a and LDLR mRNA and their correlation in terms of expression levels in porcine liver tissue were analyzed using reverse-transcription quantitative PCR. Results: Gel electrophoresis and sequencing showed that target gene fragments were successfully cloned, and the three recombinant vectors were successfully constructed. Compared to pMR-mCherry, the miR-20a expression vector significantly inhibited wild-type LDLR3'-UTR-driven (p<0.01), but not mutant LDLR-3'-UTR-driven (p>0.05), luciferase reporter activity. Further, miR-20a and LDLR were expressed at relatively high levels in porcine liver tissues. Pearson correlation analysis revealed that porcine liver miR-20a and LDLR levels were significantly negatively correlated (r = -0.656, p<0.05). Conclusion: LDLR is a potential target of miR-20a, which might directly bind the LDLR 3'-UTR to post-transcriptionally inhibit expression. These results have implications in understanding the pathogenesis and progression of porcine cardiovascular diseases.

• Development and Reproduction
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• v.12 no.3
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• pp.289-296
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• 2008

BPES (Blepharophimosis/Ptosis/Epicanthus inversus Syndrome) is an autosomal dominant disorder caused by mutations in FOXL2. Affected individuals have premature ovarian failure (POF) in addition to small palpebral fissures, drooping eyelids, and broad nasal bridge. FOXL2 is a member of the forkhead family transcription factors. In FOXL2-deficient ovaries, granulosa cell differentiation dose not progress, leading to arrest of folliculogenesis and oocytes atresia. Using yeast two-hybrid screening of rat ovarian cDNA library with FOXL2 as bait, we found that small ubiquitin-related modifier (SUMO)-conjugating E2 enzyme UBE2I protein interacted with FOXL2 protein. UBE2I also known as UBC9 is an essential protein for processing SUMO modification. Sumoylation is a form of post-translational modification involved in diverse signaling pathways including the regulation of transcriptional activities of many transcriptional factors. In the present study, we confirmed the protein-protein interaction between FOXL2 and UBE2I in human cells, 293T, by in vivo immunoprecipitation. In addition, we generated truncated FOXL2 mutants and identified the region of FOXL2 required for its association with UBE2I using yeast-two hybrid system. Therefore, the identification of UBE2I as an interacting protein of FOXL2 further suggests a presence of novel regulatory mechanism of FOXL2 by sumoylation.

• 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.

• MISULJARYO - National Museum of Korea Art Journal
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• v.96
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• pp.54-88
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• 2019

The Album of Complete Views of Seas and Mountains comprises sixty real scenery landscape paintings depicting Geumgangsan Mountain, the Haegeumgang River, and the eight scenic views of Gwandong regions, as well as fifty-one pieces of writing. It is a rare example in terms of its size and painting style. The paintings in this album, which are densely packed with natural features, follow the painting style of the Southern School yet employ crude and unconventional elements. In them, stones on the mountains are depicted both geometrically and three-dimensionally. Since 1973, parts of this album have been published in some exhibition catalogues. The entire album was opened to the public at the special exhibition "Through the Eyes of Joseon Painters: Real Scenery Landscapes of Korea" held at the National Museum of Korea in 2019. The Album of Complete Views of Seas and Mountains was attributed to Kim Eung-hwan (1742-1789) due to the signature on the final leaf of the album and the seal reading "Bokheon(painter's penname)" on the currently missing album leaf of Chilbodae Peaks. However, there is a strong possibility that this signature and seal may have been added later. This paper intends to reexamine the creator of this album based on a variety of related factors. In order to understand the production background of Album of Complete Views of Seas and Mountains, I investigated the eighteenth-century tradition of drawing scenic spots while travelling in which scenery of was depicted during private travels or official excursions. Jeong Seon(1676-1759), Sim Sa-jeong(1707-1769), Kim Yun-gyeom(1711-1775), Choe Buk(1712-after 1786), and Kang Se-hwang(1713-1791) all went on a journey to Geumgangsan Mountain, the most famous travel destination in the late Joseon period, and created paintings of the mountain, including Album of Pungak Mountain in the Sinmyo Year(1711) by Jeong Seon. These painters presented their versions of the traditional scenic spots of Inner Geumgangsan and newly depicted vistas they discovered for themselves. To commemorate their private visits, they produced paintings for their fellow travelers or sponsors in an album format that could include several scenes. While the production of paintings of private travels to Geumgangsan Mountain increased, King Jeongjo(r. 1776-1800) ordered Kim Eung-hwan and Kim Hong-do, court painters at the Dohwaseo(Royal Bureau of Painting), to paint scenic spots in the nine counties of the Yeongdong region and around Geumgangsan Mountain. King Jeongjo selected these two as the painters for the official excursion taking into account their relationship, their administrative experience as regional officials, and their distinct painting styles. Starting in the reign of King Yeongjo(r. 1724-1776), Kim Eung-hwan and Kim Hong-do served as court painters at the Dohwaseo, maintained a close relationship as a senior and a junior and as colleagues, and served as chalbang(chief in large of post stations) in the Yeongnam region. While Kim Hong-do was proficient at applying soft and delicate brushstrokes, Kim Eung-hwan was skilled at depicting the beauty of robust and luxuriant landscapes. Both painters produced about 100 scenes of original drawings over fifty days of the official excursion. Based on these original drawings, they created around seventy album leaves or handscrolls. Their paintings enriched the tradition of depicting scenic spots, particularly Outer Inner Geumgang and the eight scenic views of Gwandong around Geumgangsan Mountain during private journeys in the eighteenth century. Moreover, they newly discovered places of scenic beauty in the Outer Geungang and Yeongdong regions, establishing them as new painting themes. The Album of Complete Views of Seas and Mountains consists of four volumes. The volumes I, II include twenty-nine paintings of Inner Geumgangsan; the volume III, seventeen scenes of Outer Geumgangsan; and the volume IV, fourteen images of Maritime Geumgangsan and the eight scenic views of Gwandong. These paintings produced on silk show crowded compositions, geometrical depictions of the stones and the mountains, and distinct presentation of the rocky peaks of Geumgangsan Mountain using white and grayish-blue pigments. This album reflects the Joseon painting style of the mid- and late eighteenth century, integrating influences from Jeong Seon, Kang Se-hwang, Sim Sa-jeong, Jeong Chung-yeop(1725-after 1800), and Kim Hong-do. In particular, some paintings in the album show similarities to Kim Hong-do's Album of Famous Mountains in Korea in terms of its compositions and painterly motifs. However, "Yeongrangho Lake," "Haesanjeong Pavilion," and "Wolsongjeong Pavilion" in Kim Eung-hwan's album differ from in the version by Kim Hong-do. Thus, Kim Eung-hwan was influenced by Kim Hong-do, but produced his own distinctive album. The Album of Complete Views of Seas and Mountains includes scenery of "Jaundam Pool," "Baegundae Peak," "Viewing Birobong Peak at Anmunjeom groove," and "Baekjeongbong Peak," all of which are not depicted in other albums. In his version, Kim Eung-hwan portrayed the characteristics of the natural features in each scenic spot in a detailed and refreshing manner. Moreover, he illustrated stones on the mountains using geometric shapes and added a sense of three-dimensionality using lines and planes. Based on the painting traditions of the Southern School, he established his own characteristics. He also turned natural features into triangular or rectangular chunks. All sixty paintings in this album appear rough and unconventional, but maintain their internal consistency. Each of the fifty-one writings included in the Album of Complete Views of Seas and Mountains is followed by a painting of a scenic spot. It explains the depicted landscape, thus helping viewers to understand and appreciate the painting. Intimately linked to each painting, the related text notes information on traveling from one scenic spot to the next, the origins of the place names, geographic features, and other related information. Such encyclopedic documentation began in the early nineteenth century and was common in painting albums of Geumgangsan Mountain in the mid- nineteenth century. The text following the painting of Baekhwaam Hermitage in the Album of Complete Views of Seas and Mountains documents the reconstruction of the Baekhwaam Hermitage in 1845, which provides crucial evidence for dating the text. Therefore, the owner of the Album of Complete Views of Seas and Mountains might have written the texts or asked someone else to transcribe them in the mid- or late nineteenth century. In this paper, I have inferred the producer of the Album of Complete Views of Seas and Mountains to be Kim Eung-hwan based on the painting style and the tradition of drawing scenic spots during official trips. Moreover, its affinity with the Handscroll of Pungak Mountain created by Kim Ha-jong(1793-after 1878) after 1865 is another decisive factor in attributing the album to Kim Eung-hwan. In contrast to the Album of Famous Mountains in Korea by Kim Hong-do, the Album of Complete Views of Seas and Mountains exerted only a minor influence on other painters. The Handscroll of Pungak Mountain by Kim Ha-jong is the sole example that employs the subject matter from the Album of Complete Views of Seas and Mountains and follows its painting style. In the Handscroll of Pungak Mountain, Kim Ha-jong demonstrated a painting style completely different from that in the Album of Seas and Mountains that he produced fifty years prior in 1816 for Yi Gwang-mun, the magistrate of Chuncheon. He emphasized the idea of "scholar thoughts" by following the compositions, painterly elements, and depictions of figures in the painting manual style from Kim Eung-hwan's Album of Complete Views of Seas and Mountains. Kim Ha-jong, a member of the Gaeseong Kim clan and the eldest grandson of Kim Eung-hwan, is presumed to have appreciated the paintings depicted in the nature of Album of Complete Views of Seas and Mountains, which had been passed down within the family, and newly transformed them. Furthermore, the contents and narrative styles of Yi Yu-won's writings attached to the paintings in the Handscroll of Pungak Mountain are similar to those of the fifty-one writings in Kim Eunghwan's album. This suggests a possible influence of the inscriptions in Kim Eung-hwan's album or the original texts from which these inscriptions were quoted upon the writings in Kim Ha-jong's handscroll. However, a closer examination will be needed to determine the order of the transcription of the writings. The Album of Complete View of Seas and Mountains differs from Kim Hong-do's paintings of his official trips and other painting albums he influenced. This album is a siginificant artwork in that it broadens the understanding of the art world of Kim Eung-hwan and illustrates another layer of real scenery landscape paintings in the late eighteenth century.