• Title/Summary/Keyword: PCR (Polymerase Chain Reaction)

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Olfactory receptor (OR7D4 and OR1I1) expression in stallion testes

  • Kim, Junyoung;Jung, Youngwook;Jung, Heejun;Shakee, Muhammad;Yoon, Minjung
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.4
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    • pp.292-298
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    • 2021
  • Olfactory receptors (OR) are primarily responsible for the detection of odorant molecules. We previously demonstrated that OR7D4, an OR for androstenone, is expressed in the vomeronasal organ and olfactory epithelium tissue of stallions. Recently, the expression of OR1I1 in the human testes was reported and the possible roles of OR1I1 in the testicular cells were suggested. The objectives of this study were 1) to explore the expression of OR7D4 and OR1I1 in stallion testes, and 2) to define the specific localization of OR7D4 and OR1I1 in the testicular tissues. Stallion testicular tissue samples were used for this study. Western blot was performed to confirm the cross-reactivity of OR7D4 and OR1I1 antibody with stallion testicular tissue samples. OR7D4 and OR1I1 gene expressions were investigated using reverse transcription-polymerase chain reaction (RT-PCR) in stallion testes. Immunofluorescence was performed to investigate the expression of OR7D4 and OR1I1 in stallion testicular tissues. The protein bands for OR7D4 and OR1I1 from the testes were observed at approximately 38 kDa and 43 kDa, respectively. The mRNA of OR7D4 and OR1I1 were detected in stallion testes. Immunolabeling of OR7D4 and OR1I1 in the cytoplasm of both spermatogonia and Leydig cells was observed. In conclusion, androstenone and another odorant chemical, which is recognized by OR1I1, may play an important role in stallion testes.

Expression of miR-29a in whole Blood of Patients with Colorectal Neoplasm

  • Hwang, Dasom;Kim, Dahye;Chang, Yunhee;Hirgo, Workneh Korma;Lee, Hyeyoung
    • Biomedical Science Letters
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    • v.27 no.4
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    • pp.216-222
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    • 2021
  • Colorectal cancer (CRC) is major cancer with high incidence and mortality worldwide. It is known that most CRCs arise from precursor adenomatous polyps (APs). Recently, microRNA (miRNA) has been proposed as a biomarker for various cancers including CRC. In this study, the expression patterns of miR-29a in the whole blood (WB) of CRC, AP, and control groups were analyzed by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) to evaluate the expression level of miR-29a in patients with colorectal neoplasm (CRN) including CRC and AP. As a result, the relative expression of miR-29a was significantly decreased in the patients with CRN compared to the control group (P<0.001). The results were in agreement with previous in vitro cell studies and studies that used tissue and feces samples, suggesting that miR-29a in WB may be useful in demonstrating the status of colorectal tissue. Additionally, we divided the control group into healthy control (HC) without any colorectal symptoms and non-tumor control (NTC) with colorectal symptoms but without any CRN. And then the relative expression of miR-29a was also significantly decreased in the NTC group compared to the HC group (P<0.001). Therefore, our study revealed that miR-29a can differentiate patients with CRN from HC group, but they are also involved in the early stage of inflammatory response and cannot be specific biomarkers for CRN.

Ecophysiology of the kleptoplastidic dinoflagellate Shimiella gracilenta: I. spatiotemporal distribution in Korean coastal waters and growth and ingestion rates

  • Ok, Jin Hee;Jeong, Hae Jin;Kang, Hee Chang;Park, Sang Ah;Eom, Se Hee;You, Ji Hyun;Lee, Sung Yeon
    • ALGAE
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    • v.36 no.4
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    • pp.263-283
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    • 2021
  • To explore the ecophysiological characteristics of the kleptoplastidic dinoflagellate Shimiella gracilenta, we determined its spatiotemporal distribution in Korean coastal waters and growth and ingestion rates as a function of prey concentration. The abundance of S. gracilenta at 28 stations from 2015 to 2018 was measured using quantitative real-time polymerase chain reaction. Cells of S. gracilenta were detected at least once at all the stations and in each season, when temperature and salinity were 1.7-26.4℃ and 9.9-35.6, respectively. Moreover, among the 28 potential prey species tested, S. gracilenta SGJH1904 fed on diverse prey taxa. However, the highest abundance of S. gracilenta was only 3 cells mL-1 during the study period. The threshold Teleaulax amphioxeia concentration for S. gracilenta growth was 5,618 cells mL-1, which was much higher than the highest abundance of T. amphioxeia (667 cells mL-1). Thus, T. amphioxeia was not likely to support the growth of S. gracilenta in the field during the study period. However, the maximum specific growth and ingestion rates of S. gracilenta on T. amphioxeia, the optimal prey species, were 1.36 d-1 and 0.04 ng C predator-1 d-1, respectively. Thus, if the abundance of T. amphioxeia was much higher than 5,618 cells mL-1, the abundance of S. gracilenta could be much higher than the highest abundance observed in this study. Eurythermal and euryhaline characteristics of S. gracilenta and its ability to feed on diverse prey species and conduct kleptoplastidy are likely to be responsible for its common spatiotemporal distribution.

The Anti-Inflammatory Effect of Gabyeobda Tea in High Fat Diet-Induced Obese Mice (가볍다차(茶)가 고지방식이로 유도된 비만 마우스에서 항염증에 미치는 효과)

  • Wu, Liangliang;Lim, Soo Kyoung;Shin, Seung-Uoo;Kim, Hojun
    • Journal of Korean Medicine for Obesity Research
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    • v.22 no.1
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    • pp.11-20
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    • 2022
  • Objectives: The purpose of this study was to investigate the effects of Gabyeobda tea (GT) on anti-inflammation in ice induced high fat diet (HFD). Methods: The C57BL/6 mice fed HFD were administrated with GT once daily for 8 weeks. The changes of body weight, calorie intake levels were measured in mice. The level of serum total cholesterol, triglyceride, high density lipoprotein cholesterol, glutamic oxaloacetic transaminase (GOT), and glutamic pyruvic transaminase (GPT) were measured in mice by enzyme-based assay. It was also observed the histological changes of liver, and fat tissues with hematoxylin and eosin staining. Further real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay were employed to detect inflammatory cytokine levels such as tumor necrosis factor (TNF)-𝛼, interleukin (IL)-6, and IL-1𝛽. Results: HFD+GT group, which was administered with GT with HFD, showed no body weight gain compared with HFD group. However, levels of GOT, GPT, and inflammatory cytokines such as TNF-𝛼, IL-6, and IL-1𝛽 in the blood of HFD+GT group were significantly reduced compared with HFD group. In addition, the messenger RNA (mRNA) expression level of the IL-12 gene was significantly reduced and the mRNA expression level of the IL-10 was increased in the liver. Conclusions: It suggests that Gabyeobda tea can alleviate inflammatory responses induced by high fat diet by inhibiting inflammatory cytokines production.

Study on the Effect of Cimicifuga heracleifolia Ethanol Extract on Hyaluronic Acid Synthesis

  • Son, Hyun-Kyu;Ha, Hun-Yong
    • Applied Chemistry for Engineering
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    • v.33 no.6
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    • pp.557-562
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    • 2022
  • Hyaluronic acid (HA) is a mucopolysaccharide, occurring naturally in living organisms. It is one of the most hydrophilic molecules, so it has been known as being related to skin hydration and skin aging. The purpose of this study was to examine the effects of Cimicifuga heracleifolia ethanol extract on the hyaluronic acid synthesis and the inhibition of hyaluronidase activity. To determine cytotoxicity, hyaluronic acid synthase 2 (HAS2) gene expression, HA production and, hyaluronidase inhibitory effects, 3-(4,5-dimethylthiazol-2-ly)-2,5-diphenyl tetrazolium bromide (MTT) assay, real time - polymerase chain reaction (RT-PCR), hyaluronic acid enzyme linked immunosorbent assay (HA-ELISA), and hyaluronidase assay were used, respectively. When the Cimicifuga heracleifolia extract was treated in the HaCaT cells up to 500 ㎍/mL concentration, cytotoxicity was confirmed by the Cimicifuga heracleifolia extract at concentrations above 200 ㎍/mL. Therefore, the optimum concentration of all experiments used in this study was determined to be 200 ㎍/mL. HAS2 gene expression increased by Cimicifuga heracleifolia extract in a concentration-dependent manner at all treatment concentrations. The production rate of HA was tended to decrease at the highest concentration of 200 ㎍/mL. The hyaluronidase activity inhibition effect of Cimicifuga heracleifolia extract was very high compared to the control group. Based on these results, Cimicifuga heracleifolia extract was expected to have a moisturizing effect on human skin and special attention should be paid to the determination of the concentration of Cimicifuga heracleifolia when developing cosmetic materials using it.

CircCOL1A2 Sponges MiR-1286 to Promote Cell Invasion and Migration of Gastric Cancer by Elevating Expression of USP10 to Downregulate RFC2 Ubiquitination Level

  • Li, Hang;Chai, Lixin;Ding, Zujun;He, Huabo
    • Journal of Microbiology and Biotechnology
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    • v.32 no.7
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    • pp.938-948
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    • 2022
  • Gastric cancers (GC) are generally malignant tumors, occurring with high incidence and threatening public health around the world. Circular RNAs (circRNAs) play crucial roles in modulating various cancers, including GC. However, the functions of circRNAs and their regulatory mechanism in colorectal cancer (CRC) remain largely unknown. This study focuses on both the role of circCOL1A2 in CRC progression as well as its downstream molecular mechanism. Quantitative polymerase chain reaction (qPCR) and western blot were adopted for gene expression analysis. Functional experiments were performed to study the biological functions. Fluorescence in situ hybridization (FISH) and subcellular fraction assays were employed to detect the subcellular distribution. Luciferase reporter, RNA-binding protein immunoprecipitation (RIP), co-immunoprecipitation (Co-IP), RNA pull-down, and immunofluorescence (IF) and immunoprecipitation (IP) assays were used to explore the underlying mechanisms. Our results found circCOL1A2 to be not only upregulated in GC cells, but that it also propels the migration and invasion of GC cells. CircCOL1A2 functions as a competing endogenous RNA (ceRNA) by sequestering microRNA-1286 (miR-1286) to modulate ubiquitin-specific peptidase 10 (USP10), which in turn spurs the migration and invasion of GC cells by regulating RFC2. In sum, CircCOL1A2 sponges miR-1286 to promote cell invasion and migration of GC by elevating the expression of USP10 to downregulate the level of RFC2 ubiquitination. Our study offers a potential novel target for the early diagnosis and treatment of GC.

Comparison of O-serogroups, Virulence Factors and Phylogenetic Groups of Uropathogenic Escherichia coli Isolated from Patients with Urinary Tract Infections between 2 Time Periods of 1989 and 2010-2014 at Gangwon Province in Korea

  • Park, Min;Kim, Seong-Mi
    • Biomedical Science Letters
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    • v.28 no.2
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    • pp.127-136
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    • 2022
  • Uropathogenic Escherichia coli (UPEC) is main causative agent of urinary tract infections. They are classified based on various types of O antigen. UPEC strains commonly possess many genes encoding virulece-associated factors. E. coli strains are generally divided into four main phylogenetic groups. The virulence factor (VF) profiles of UPEC are related with their O-serogroups in each strains. A total of 681 strains of UPEC clinical isolates were collected from Korean healthcare facility (1989: 123 strains and 2010-2014: 558 strains). The UPEC clinical isolates were analyzed by polymerase chain reaction (PCR) methods. A total of 14 O-serotypes (O1, O2, O4, O6, O7, O8, O15, O16, O18, O21, O22, O25, O75 and O83), 6 virulence factors (papC, fimG/H, sfaD/E, hly1, cnf1 and usp) and phylogenetic groups were identified. The most prevalent O-serogroups were O6 (11.1%) in 1989 UPEC strains and O25 (21.0%) in 2010-2014 UPEC strains. The identified VFs, phylogenetic groups in 1989 UPEC strains and 2010-2014 UPEC strains were fimG/H and B2 group. In this study, O6 serotype was revealed the close relationships with VFs. Also, the distribution of prevalence O-serogroups of UPEC has been changed from O6 to O25 and virulence of UPEC strains was increased during past twenty-one years.

Two Clinical Cases of Feline Hemoplasmosis in Korea

  • Kim, Young Ju;Bae, Hyeona;Shin, Sun Woo;Cho, ARom;Jeon, Yeseul;Hwang, Tae-Sung;Jung, Dong-In;Kim, Dae Young;Kang, Jun-Gu;Yu, DoHyeon
    • Parasites, Hosts and Diseases
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    • v.60 no.2
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    • pp.127-131
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    • 2022
  • Feline hemotropic mycoplasmosis (hemoplasmosis) is an infection of the red blood cells caused by the Mycoplasma haemofelis (Mhf), Candidatus Mycoplasma haemominutum (CMhm), and Candidatus Mycoplasma turicensis (CMt). The existence of Mhf, CMhm, and CMt has been demonstrated in feral cats in Korea using molecular methods, but no clinical cases have yet been reported. This study reports 2 clinical cases of hemotropic mycoplasmosis caused by CMhm and CMt in 2 anemic cats. The first case was a client-owned intact female domestic shorthair cat that presented with fever, pale mucous membranes, and normocytic normochromic non-regenerative anemia. Prior to referral, an immunosuppressive prednisolone dose was administered at the local veterinary clinic for 1 month. The cat was diagnosed with high-grade alimentary lymphoma. Organisms were found on the surface of the red blood cells on blood smear examination. The second case was of a rescued cat that presented with dehydration and fever. The cat had normocytic normochromic non-regenerative anemia. Necropsy revealed concurrent feline infectious peritonitis. Polymerase chain reaction assay targeting 16S rRNA revealed CMhm infection in case 1 and dual infection of CMhm and CMt in case 2. Normocytic normochromic non-regenerative anemia was observed in both cats before and during the management of the systemic inflammation. This is the first clinical case report in Korea to demonstrate CMhm and CMt infections in symptomatic cats.

Betulin, an Anti-Inflammatory Triterpenoid Compound, Regulates MUC5AC Mucin Gene Expression through NF-kB Signaling in Human Airway Epithelial Cells

  • Hossain, Rajib;Kim, Kyung-il;Jin, Fengri;Lee, Hyun Jae;Lee, Choong Jae
    • Biomolecules & Therapeutics
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    • v.30 no.6
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    • pp.540-545
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    • 2022
  • Betulin is a triterpenoid natural product contained in several medicinal plants including Betulae Cortex. These medicinal plants have been used for controlling diverse inflammatory diseases in folk medicine and betulin showed anti-inflammatory, antioxidative, and anticancer activities. In this study, we tried to examine whether betulin exerts a regulative effect on the gene expression of MUC5AC mucin under the status simulating a pulmonary inflammation, in human airway epithelial cells. Confluent NCI-H292 cells were pretreated with betulin for 30 min and then stimulated with phorbol 12-myristate 13-acetate (PMA) for 24 h or the indicated periods. The MUC5AC mucin mRNA expression and mucin glycoprotein production were measured by reverse transcription - polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. To elucidate the action mechanism of betulin, effect of betulin on PMA-induced nuclear factor kappa B (NF-kB) signaling pathway was also investigated by western blot analysis. The results were as follows: 1) Betulin significantly suppressed the production of MUC5AC mucin glycoprotein and down-regulated MUC5AC mRNA expression induced by PMA in NCI-H292 cells. 2) Betulin inhibited NF-κB activation stimulated by PMA. Suppression of inhibitory kappa B kinase (IKK) by betulin led to the inhibition of the phosphorylation and degradation of inhibitory kappa B alpha (IκBα), and the nuclear translocation of NF-κB p65. This, in turn, led to the down-regulation of MUC5AC glycoprotein production in NCI-H292 cells. These results suggest betulin inhibits the gene expression of mucin through regulation of NF-kB signaling pathway, in human airway epithelial cells.

Molecular identification and characterization of Lumpy skin disease virus emergence from cattle in the northeastern part of Thailand

  • Seerintra, Tossapol;Saraphol, Bhuripit;Wankaew, Sitthichai;Piratae, Supawadee
    • Journal of Veterinary Science
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    • v.23 no.5
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    • pp.73.1-73.8
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    • 2022
  • Background: Lumpy skin disease (LSD), a disease transmitted by direct and indirect contact with infected cattle, is caused by the Lumpy skin disease virus (LSDV). The disease affects cattle herds in Africa, Europe, and Asia. The clinical signs of LSD range from mild to the appearance of nodules and lesions in the skin leading to severe symptoms that are sometimes fatal with significant livestock economic losses. Objectives: This study aimed to characterize LSDV strains in the blood of infected cattle in Thailand based on the GPCR gene and determine the phylogenetic relationship of LSDV Thailand isolates with published sequences available in the database. Methods: In total, the blood samples of 120 cattle were collected from different farms in four provinces in the northeastern part of Thailand, and the occurrence of LSDV was examined by PCR based on the P32 antigen gene. The genetic diversity of LSDV based on the GPCR gene was analyzed. Results: Polymerase chain reaction assays based on the P32 antigen gene showed that 4.17% (5/120) were positive for LSDV. All positive blood samples were amplified successfully for the GPCR gene. Phylogenetic analysis showed that LSDV Thailand isolates clustered together with LSDVs from China and Russia. Conclusions: The LSD outbreak in Thailand was confirmed, and a phylogenetic tree was constructed to infer the branching pattern of the GPCR gene from the presence of LSDV in Thailand. This is the first report on the molecular characterization of LSDV in cattle in Thailand.