Association of Microbial Dysbiosis with Gallbladder Diseases Identified by Bile Microbiome Profiling

Choi Seong Ji,Kim Yeseul,Jeon Jehyun,Gwak Ho-Jin,Kim Mimi,Kang Kyojin,Kim Yohan,Jeong Jaemin,Jung Yun Kyung,Lee Kyeong Geun,Choi Ho Soon,Jung Dong-Hwan,Lee Sung-Gyu,Lee Yangsoon,Shin Su-Jin,Jang Kiseo 대한의학회 2021 Journal of Korean medical science Vol.36 No.28

Background: Cholecystitis is an important risk factor for gallbladder cancer, but the bile microbiome and its association with gallbladder disease has not been investigated fully. We aimed to analyze the bile microbiome in normal conditions, chronic cholecystitis, and gallbladder cancer, and to identify candidate bacteria that play an important role in gallbladder carcinogenesis. Methods: We performed metagenome sequencing on bile samples of 10 healthy individuals, 10 patients with chronic cholecystitis, and 5 patients with gallbladder cancer, and compared the clinical, radiological, and pathological characteristics of the participants. Results: No significant bacterial signal was identified in the normal bile. The predominant dysbiotic bacteria in both chronic cholecystitis and gallbladder cancer were those belonging to the Enterobacteriaceae family. Klebsiella increased significantly in the order of normal, chronic cholecystitis, and gallbladder cancer. Patients with chronic cholecystitis and dysbiotic microbiome patterns had larger gallstones and showed marked epithelial atypia, which are considered as precancerous conditions. Conclusion: We investigated the bile microbiome in normal, chronic cholecystitis, and gallbladder cancer. We suggest possible roles of Enterobacteriaceae, including Klebsiella, in gallbladder carcinogenesis. Our findings reveal a possible link between a dysbiotic bile microbiome and the development of chronic calculous cholecystitis and gallbladder cancer.

Expression and Localization of Na+/K+-ATPase Subunits, ATP1A1 and ATP1B1, in the Uterine Endometrium at the Maternal-Fetal Interface in Pigs

Yohan Choi,Heewon Seo,Jangsoo Shim,Mingoo Kim,Hakhyun Ka 한국동물생명공학회(구 한국동물번식학회) 2011 발생공학 국제심포지엄 및 학술대회 Vol.2011 No.1

Na+/K+-ATPase, an energy-transducing ion pump, is responsible for maintenance of relatively high concentrations of potassium ions but low concentrations of sodium ions in the cell by transport of these ions across the plasma membrane. Na+/K+-ATPase consists of α, β, and γ subunits, but only α and β subunits are needed for basic functions. Na+/K+-ATPase is also involved in regulation of intracellular calcium ion concentration by coupling with Na+/Ca2+ exchanger involved in intracellular calcium extrusion. Our previous study showed that calcium regulatory molecules including Na+/Ca2+ exchanger are expressed in the uterine endometrium during the estrous cycle and pregnancy in pigs, however, expression of Na+/K+-ATPase in the uterine endometrium has not been determined. Thus, we examined expression of α1 (ATP1A1) and β1 (ATP1- B1) subunits of Na+/K+-ATPase in the uterine endometrium during the estrous cycle and pregnancy in pigs. Real-time RT-PCR analysis showed that levels of ATP1A1 m- RNA in the uterine endometrium during the estrous cycle and early pregnancy were higher than those during mid and term pregnancy, and that levels of ATP1B1 mRNA were highest on day (D) 12 of the estrous cycle. In situ hybridization analysis revealed that ATP1A1 and ATP1B1 mRNAs were localized to luminal (LE) and glandular epithelia (GE) in the endometrium. During mid to term pregnancy, localization of ATP1A1 mRNA was confined to LE, GE, and chorionic membrane (CM) of areolae and ATP1- B1 mRNA was localized to LE, GE and CM with the strongest intensity in LE of areolae. Signal intensity of ATP1B1 mRNA in LE was slightly stronger than that in GE. RT-PCR analysis showed that ATP1A1 and ATP1B1 mRNAs were expressed in conceptuses on D12 and D15 of pregnancy. These results showed that ATP1A1 and ATP1B1 were expressed in the uterine endometrium and conceptuses during the estrous cycle and pregnancy in a pregnancy status- and stage-specific manner. These suggest that Na+/K+-ATPase may play a key role in the establishment and maintenance of pregnancy by regulating intracellular concentrations of various ions including calcium at the maternal-fetal interface in pigs.

Benign Colorectal Disease 1

Yohan Choi 대한외과학회 2016 대한외과학회 학술대회 초록집 Vol.2016 No.11

Analysis of Na+/K+-ATPase in the uterine endometrium during the estrous cycle and pregnancy in pigs

Yohan Choi,Heewon Seo,Jangsoo Shim,Inkyu Yoo,Hakhyun Ka 한국발생생물학회 2013 한국발생생물학회 학술발표대회 Vol.2013 No.8

Na+/K+-ATPase, an energy-transducing ion pump, is responsible for maintenance of relatively high concentrations of potassium ions but low concentrations of sodium ions in the cell by transport of these ions across the plasma membrane and participates in transport of various nutrients including glucose, amino acids, and ions. In addition, Na+/K+-ATPase is also involved in regulation of intracellular calcium ion concentration by coupling with Na+/Ca+ exchanger expressed at the maternal-fetal interface in pigs. Na+/K+-ATPase consists of α, β, and FXYD subunits, but only α and β subunits are required for primary functions. FXYD subunit is an auxiliary protein for αβ complex of Na+/K+-ATPase. However, it has not been determined that subunits of Na+/K+-ATPase are expressed in the uterine endometrium during the estrous cycle and pregnancy in pigs. In this study, we determined expression of alpha (ATP1A1-4), beta (ATP1B1-3), and FXYD (FXYD1-7) subunits of Na+/K+-ATPase in the uterine endometrium during the estrous cycle and pregnancy in pigs. Real-time RT-PCR analysis showed that all alpha, beta, and FXYD subunits, except ATP1A3, were expressed in the uterine endometrium during the estrous cycle and pregnancy in a pregnancy status- and stage-specific fashion. In situ hybridization analysis exhibited that ATP1A1, ATP1A4, and ATP1B1 were localized to luminal (LE) and glandular epithelium (GE) during the estrous cycle and early pregnancy, and during mid to term pregnancy. ATP1A1 mRNA was localized to LE, GE, and areolae of the chorioallantois, especially at high levels to LE in areolae regions. ATP1B3 mRNA was detected only in LE during the estrous cycle and pregnancy with highest levels on day (D) 12 of pregnancy. Transcripts of all subtypes of FXYD subunit were primarily localized to LE and GE during the estrous cycle and early pregnancy and to chorionic membrane (CM) during mid to term pregnancy. RT-PCR analysis showed that all subtypes of Na+/K+-ATPase subunits, except ATP1A2, ATP1A3, and ATP1B2 mRNAs, were expressed in conceptuses on D12 and D15 of pregnancy. These results indicate that Na+/K+-ATPase subunits are expressed in the uterine endometrium and conceptuses during the estrous cycle and pregnancy in a pregnancy status- and stage-specific manner. These suggest that Na+/K+-ATPase subunits may be involved in the establishment and maintenance of pregnancy by coordinate regulation of absorption and secretion of nutrients such as glucose, amino acids, and ions at the maternal-fetal interface in pigs.

Maternal-Conceptus Interactions: Mediators Regulating the Implantation Process in Pigs

Choi, Yohan,Seo, Heewon,Yoo, Inkyu,Han, Jisoo,Jang, Hwanhee,Kim, Minjeong,Ka, Hakhyun The Korean Society of Animal Reproduction 2014 Reproductive & developmental biology Vol.38 No.1

For successful embryo implantation, the communication of the maternal endometrium with the conceptus trophectoderm is required essentially. In pigs, conceptuses undergo morphological change in length to enlarge the physical contact area with the maternal endometrium and secrete estrogen to induce the maternal recognition of pregnancy during the peri-implantation period. Conceptus-derived estrogen prevents luteolysis by conversion in direction of $PGF_{2{\alpha}}$ secretion from the uterine vasculature to the uterine lumen as well as it affects on expression of the uterine endometrial genes. In addition to estrogen, conceptuses release various signaling molecules, including cytokines, growth factors, and proteases, and, in response to these signaling molecules, the maternal uterine endometrium also synthesizes many signaling molecules, including hormones, cytokines, growth factors, lipid molecules, and utilizes ions such as calcium ion by calcium regulatory molecules. These reciprocal interactions of the conceptus trophectoderm with the maternal uterine endometrium make development and successful implantation of embryos possible. Thus, signaling molecules at the maternal-conceptus interface may play an important role in the implantation process. This review summarized syntheses and functions of signaling molecules at the maternal-conceptus interface to further understand mechanisms of the embryo implantation process in pigs.

Expression of TRPV6 in the Uterine Endometrium during the Estrous Cycle and Pregnancy in Pig and Regulation of Its Expression by Steroid Hormones

Yohan Choi,Heewon Seo,Hakhyun Ka 한국동물생명공학회(구 한국동물번식학회) 2007 발생공학 국제심포지엄 및 학술대회 Vol.2007 No.1

Decorin Expression in the Uterine Endometrium during the Estrous Cycle and Pregnancy in Pigs

Yohan Choi,Heewon Seo,Mingoo Kim,Hakhyun Ka 한국동물생명공학회(구 한국동물번식학회) 2009 발생공학 국제심포지엄 및 학술대회 Vol.2009 No.1

Night feeding in lactating sows is an essential management approach to decrease the detrimental impacts of heat stress

( Yohan Choi ),( Joseph Moturi ),( Abdolreza Hosseindoust ),( Minju Kim ),( Kwangyeol Kim ),( Junhyung Lee ),( Changhyun Song ),( Younghwa Kim ),( Byungjo Chae ) 한국축산학회 2019 한국축산학회지 Vol.61 No.6

The present study investigated the litter performance of multiparous sows had different feeding frequencies during summer season. A total of 60 crossbred multiparous sows was allotted to one of two treatments based on body weight (BW) according to a completely randomized design. Two different feeding frequencies based on NRC (National Research Council) were applied as conventional feeding (T1; 2 kg per meal at 06:00, 11:00, and 17:00) and night feeding (T2; 1.5 kg per meal at 06:00, 11:00, 17:00, and 2 kg at 22:00). Sows in T2 treatment had lower (p < 0.05) BW changes during lactation. The backfat thickness change of sows was decreased in T2 treatment during lactation (p < 0.05). The daily feed intake differed significantly between T1 and T2 with increased feed intake of the T2 group at 5.47 kg/d to 5.14 kg/d in the T1 group (p < 0.05). There was a significant difference in total weight of weaned piglets between T1 (70.50 kg) and T2 (74.34 kg). A greater total litter weight gain was observed in sows in T2 treatment. Night feeding is suggested for lactating sows with significant beneficial effects on litter growth during summer season.

Klotho : Expression and Regulation at the Maternal-Conceptus Interface in Pigs

Yohan Choi,Heewon Seo,Jangsoo Shim,Sang-Hwan Hyun,Eunsong Lee,Hakhyun Ka 한국수정란이식학회 2014 한국동물생명공학회지 Vol.29 No.4

Klotho (KL) is a single transmembrane protein composed of KL1 and KL2 repeats possessing β-glucuronidase activity and maintains calcium homeostasis in physiological state. It has been implicated in pigs that calcium is important for the establishment and maintenance of pregnancy, and our previous study has shown that transient receptor potential vanilloid type 6 (TRPV6), a calcium ion transporter, is predominantly expressed in the uterine endometrium during pregnancy in pigs. However, expression and function of KL in the uterine endometrium has not been determined in pigs. Thus, the present study determined expression and regulation of KL in the uterine endometrium during the estrous cycle and pregnancy in pigs. Real-time RT-PCR analysis showed that levels of KL mRNA decreased between Days 12 to 15 of the estrous cycle, and its expression showed a biphasic manner during pregnancy. KL mRNA was expressed in conceptuses and in chorioallantoic tissues during pregnancy. Explant culture study showed that expression levels of KL were not affected by treatment of steroid hormones or interleukin-1beta during the implantation period. Furthermore, levels of KL mRNA in the uterine endometrium from gilts carrying somatic cell nuclear transfer (SCNT)- derived embryos were significantly lower than those from gilts carrying natural mating-derived embryos on Day 12 of pregnancy. These results exhibited that KL was expressed at the maternal-conceptus interface in a pregnancy statusand stage-specific manner, and its expression was affected by SCNT procedure, suggesting that KL may play an important role in the establishment and maintenance of pregnancy in pigs.

Effects of Ecklonia cava as fucoidan-rich algae on growth performance, nutrient digestibility, intestinal morphology and caecal microflora in weanling pigs

Choi, Yohan,Hosseindoust, Abdolreza,Goel, Akshat,Lee, Suhyup,Jha, Pawan Kumar,Kwon, Ill Kyong,Chae, Byung-Jo Asian Australasian Association of Animal Productio 2017 Animal Bioscience Vol.30 No.1

Objective: In the present study, role of increasing levels of Ecklonia cava (seaweed) supplementation in diets was investigated on growth performance, coefficient of total tract apparent digestibility (CTTAD) of nutrients, serum immunoglobulins, cecal microflora and intestinal morphology of weanling pigs. Methods: A total of 200 weaned pigs (Landrace${\times}$Yorkshire${\times}$Duroc; initial body weight $7.08{\pm}0.15kg$) were randomly allotted to 4 treatments on the basis of body weight. There were 5 replicate pens in each treatment including 10 pigs of each. Treatments were divided by dietary Ecklonia cava supplementation levels (0%, 0.05%, 0.1%, or 0.15%) in growing-finishing diets. There were 2 diet formulation phases throughout the experiment. The pigs were offered the diets ad libitum for the entire period of experiment in meal form. Results: The pigs fed with increasing dietary concentrations of Ecklonia cava had linear increase (p<0.05) in the overall average daily gain, however, there were no significant differences in gain to feed ratio, CTTAD of dry matter and crude protein at both phase I and phase II. Digestibility of gross energy was linearly improved (p<0.05) in phase II. At day 28, pigs fed Ecklonia cava had greater (linear, p<0.05) Lactobacillus spp., fewer Escherichia coli (E. coli) spp. (linear, p<0.05) and a tendency to have fewer cecal Clostridium spp. (p = 0.077). The total anaerobic bacteria were not affected with supplementation of Ecklonia cava in diets. Polynomial contrasts analysis revealed that villus height of the ileum exhibited a linear increase (p<0.05) in response with the increase in the level of dietary Ecklonia cava. However, villus height of duodenum and jejunum, crypt depth, villus height to crypt depth ratio of different segments of the intestine were not affected. Conclusion: The results suggest that Ecklonia cava had beneficial effects on the growth performance, cecal microflora, and intestinal morphology of weanling pigs.