• 제목/요약/키워드: molecular analyses

검색결과 1,564건 처리시간 0.029초

Analysis of Bovine Interferon-tau Gene subtypes Expression in the Trophoblast and Non-trophoblast cells

  • Kim, Min-Su;Lim, Hyun-Joo;Lee, Ji Hwan;Park, Soo Bong;Won, Jeong-Il;Kim, Hyun Jong
    • 한국수정란이식학회지
    • /
    • 제33권4호
    • /
    • pp.195-203
    • /
    • 2018
  • Interferon-tau (IFNT) is known as a major conceptus protein that signals the process of maternal recognition of pregnancy in ruminants. Also, multiple interferon genes exist in cattle, However, molecular mechanisms of these bovine IFNT (bIFNT) genes whose expressions are limited have not been characterized. We and others have observed that expression levels of bovine subtype IFNT genes in the tissues of ruminants; thus, bIFNT1 and other new type I (bIFNTc1/c2/c3) gene co-exist during the early stages of conceptus development and non-trophoblast cells. Its genes transcription could be regulated through CDX2 and ETS2 and JUN and/or cAMP-response element binding protein (CREB)-binding protein (CREBBP) expression, a transcription factor implicated in the control of cell differentiation in the trophectoderm. Bovine ear-derived fibroblast cells, were co-transfected with luciferase reporter constructs carrying upstream (positions -1000 to +51) regions of bIFNT1 and other new type I gene and various transcription factor expression plasmids. Compared to each - 1kb-bIFNT1/c1/c2/c3-Luc increased when this constructs were co-transfected with CDX2, ETS2, JUN and/or CREBBP. Also, Its genes was had very effect on activity by CDX2, either alone or with the other transcription factors, markedly increased luciferase activity. However, the degree of transcriptional activation of the bIFNTc1 gene was not similar to that bIFNT1/c2/c3 gene by expression plasmid. Furthermore, Sequence analyses also revealed that the expression levels of bIFNT1/c2/c3 gene mRNAs expression were highest on day 17, 20 and 22 trophoblast and, Madin-Darby bovine kidney (MDBK), Bovine ear-derived fibroblast (EF), and endometrium (Endo) non-trophoblast cells. But, bIFNTc1 mRNA had not same expression level, bIFNTc1 lowest levels than those of IFNT1/c2/c3 gene in both trophoblast and non-trophoblast cells. These results demonstrate that bovine subtype bIFNT genes display differential, in the trophoblast and non-trophoblast cells.

Combination Therapy with a PI3K/mTOR Dual Inhibitor and Chloroquine Enhances Synergistic Apoptotic Cell Death in Epstein-Barr Virus-Infected Gastric Cancer Cells

  • Kim, Mi-Young;Kruger, Annie J.;Jeong, Ju-Yeon;Kim, Jaehee;Shin, Phil kyung;Kim, Sun Young;Cho, Joo Young;Hahm, Ki Baik;Hong, Sung Pyo
    • Molecules and Cells
    • /
    • 제42권6호
    • /
    • pp.448-459
    • /
    • 2019
  • The phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) signaling pathway is a promising target for gastric cancer (GC) treatment; however the efficacy of PI3K/mTOR dual inhibitors in GC has not yet been maximized. Additionally, the effect of autophagy regulation by PI3K/mTOR dual inhibitors has not been clearly elucidated in GC treatment. We aimed to show that our newly developed PI3K/mTOR dual inhibitor, CMG002, when combined with an autophagy inhibitor, chloroquine (CQ), potently induces effective cancer cell death in Epstein-Barr virus (EBV)-associated gastric cancer (EBVaGC) cells, where both the PI3K/AKT/mTOR and autophagy pathways play important roles in disease pathogenesis. EBV- and mock-infected AGS and NUGC3 GC cell lines were treated with CMG002 +/- CQ. PI3K/AKT/mTOR signaling pathway mediators, cellular apoptosis and autophagy markers were confirmed by Western blot assay. Cell viability was assessed using the Cell Counting Kit-8 (CCK-8) assay. CMG002 effectively blocked the PI3K/AKT/mTOR pathway by markedly decreasing phosphorylation of AKT and its downstream mediator S6. CMG002 induced G0/G1 cell cycle arrest and enhanced apoptotic cell death in AGS and NUGC3 cells, particularly EBV-infected cells compared with mock-infected cells, as confirmed by flow cytometric analyses and TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assays. The combination of CMG002 plus CQ synergistically increased apoptotic cell death in EBV-infected GC cell lines when compared with CMG002 alone (P < 0.05). Our results suggest that the new PI3K/mTOR dual inhibitor, CMG002, when used in combination with the autophagy inhibitor, CQ, provides enhanced therapeutic efficacy against EBVaGC.

토양-지하수내 미세플라스틱 거동 연구를 위한 수정진동자미세저울 기술 소개 (Application of Quartz Crystal Microbalance to Understanding the Transport of Microplastics in Soil and Groundwater)

  • 김주혁;명현아;손상보;권기덕
    • 광물과 암석
    • /
    • 제33권4호
    • /
    • pp.463-475
    • /
    • 2020
  • 최근 토양과 지하수에서도 미세플라스틱이 발견되어 미세플라스틱 환경오염 관련 연구의 중요성이 크게 대두되고 있다. 주로 ㎛ - nm의 작은 입자로 존재하는 점토광물과 금속산화광물은 표면적이 넓어 미세플라스틱에 대한 흡착력 등 화학 반응도가 매우 높기 때문에, 광물표면 상호작용은 토양과 지하수 환경 내 미세플라스틱의 거동을 결정하는 중요한 역할을 할 수 있다. 따라서, 광물과 미세플라스틱 간의 상호작용에 대한 환경광물학 연구는 미세플라스틱 거동 예측 기술개발 및 오염대책 마련에 핵심이 되는 연구분야라 할 수 있다. 광물표면과 미세플라스틱(특히, 나노플라스틱) 연구에는 분자-나노수준의 분석기술이 요구된다. 이번 기술보고에서는 나노그람(=10-9 g) 수준의 질량 변화를 실시간으로 측정할 수 있는 초정밀 분석기기로, 광물표면에 흡·탈착되는 미세플라스틱 및 나노플라스틱의 미세한 질량 변화를 측정할 수 있는, 수정진동자미세저울(quartz crystal microbalance, QCM)을 소개한다. QCM 작동원리를 소개하고, 대표적인 QCM 연구결과와 기존 컬럼 실험과의 장단점을 비교하여 미세플라스틱 연구에 QCM 활용 가능성을 논의한다.

Morphological and Molecular Identification of Spirometra Tapeworms (Cestoda: Diphyllobothriidae) from Carnivorous Mammals in the Serengeti and Selous Ecosystems of Tanzania

  • Ndosi, Barakaeli Abdieli;Park, Hansol;Lee, Dongmin;Choe, Seongjun;Kang, Yeseul;Nath, Tilak Chandra;Bia, Mohammed Mebarek;Eamudomkarn, Chatanun;Jeon, Hyeong-Kyu;Eom, Keeseon S.
    • Parasites, Hosts and Diseases
    • /
    • 제58권6호
    • /
    • pp.653-660
    • /
    • 2020
  • Spirometra tapeworms (Cestoda: Diphyllobothriidae) collected from carnivorous mammals in Tanzania were identified by the DNA sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and internal transcribed spacer 1 (ITS1), and by morphological characteristics. A total of 15 adult worms were collected from stool samples and carcasses of Panthera leo, Panthera pardus, and Crocuta crocuta in the Serengeti and Selous ecosystems of Tanzania. Three Spirometra species: S. theileri, S. ranarum and S. erinaceieuropaei were identified based on morphological features. Partial cox1 sequences (400 bp) of 10 specimens were revealed. Eight specimens showed 99.5% similarity with Spirometra theileri (MK955901), 1 specimen showed 99.5% similarity with the Korean S. erinaceieuropaei and 1 specimen had 99.5% similarity with Myanmar S. ranarum. Sequence homology estimates for the ITS1 region of S. theileri were 89.8% with S. erinaceieuropaei, 82.5% with S. decipiens, and 78.3% with S. ranarum; and 94.4% homology was observed between S. decipiens and S. ranarum. Phylogenetic analyses were performed with 4 species of Spirometra and 2 species of Dibothriocephalus (=Diphyllobothrium). By both ML and BI methods, cox1 and ITS1 gave well supported, congruent trees topology of S. erinaceieuropaei and S. theileri with S. decipiens and S. ranarum forming a clade. The Dibothriocephalus species were sisters of each other and collectively forming successive outgroups. Our findings confirmed that 3 Spirometra species (S. theileri, S. ranarum, and S. erinaceieuropaei) are distributed in the Serengeti and Selous ecosystems of Tanzania.

Surveillance on the Vivax Malaria in Endemic Areas in the Republic of Korea Based on Molecular and Serological Analyses

  • Lee, Seong-Kyun;Hu, Fengyue;Firdaus, Egy Rahman;Park, Ji-Hoon;Han, Jin-Hee;Lee, Sang-Eun;Shin, Hyun-Il;Cho, Shin Hyeong;Park, Won Sun;Lu, Feng;Han, Eun-Taek
    • Parasites, Hosts and Diseases
    • /
    • 제58권6호
    • /
    • pp.609-617
    • /
    • 2020
  • Plasmodium vivax reemerged in 1993. It has been sustained for more than 25 years and become one of the important indigenous parasitic diseases in northern and western parts of the Republic of Korea near the demilitarized zone. In particular, relapse is a significant concern for the control of malaria, as short- and long-term incubation periods vary among those infected in Korea. In this study, the prevalence of asymptomatic carriers was examined among residents of high endemic areas of vivax malaria during nonseasonal transmission of mosquitoes. Blood samples from 3 endemic regions in northwestern Korea were evaluated by microscopic examination, rapid diagnostic testing, and nested PCR to identify asymptomatic patients carrying malaria parasites in the community. However, no positive malaria case among residents of endemic areas was detected. Additionally, serological analysis was carried out to measure antibodies against 3 antigenic recombinant proteins of P. vivax, merozoite surface protein 1-19, circumsporozoite surface protein-VK210, and liver-stage antigen (PvLSA-N), by the protein array method. Interestingly, seropositivity of sera between previous exposure and samples without exposure to malaria was significantly higher using the PvLSA-N antigen than the other antigens, suggesting that PvLSA-N can be used as a serological marker to analyze the degree of exposure for malaria transmission in endemic areas. This indicates a very low asymptomatic carrier prevalence during the nonmalaria season in the endemic areas of Korea.

토란으로부터 분리한 면역활성 다당 galactomannan의 구조적 특성 규명 (Structural elucidation of immuno-stimulating polysaccharide, galactomannan isolated from Colocasia esculenta)

  • 이희원;신광순
    • 한국식품과학회지
    • /
    • 제52권6호
    • /
    • pp.595-603
    • /
    • 2020
  • 토란에 존재하는 점질다당의 새로운 이용방안을 모색하기 위하여 토란으로부터 다당을 분리하여 항보체 활성을 평가하고 구조 분석을 행하였다. 토란으로부터 분리한 조다당 CE를 이온교환수지와 Sephadex G-100 column를 이용하여 정제하였고, 그 중 수율과 활성이 양호한 CE-4a를 최종 획분으로 선정하였다. 초기 면역반응에 중요한 역할을 하는 보체계에 대한 토란 다당의 활성화 여부를 측정한 결과 양성대조군인 PSK에 준하는 강력한 항보체 활성을 보였고, 시료의 농도차이를 두어 실험한 결과 농도 의존적임을 알 수 있었다. CE-4a는 분자량 약 182.4 kDa의 다당체로 구성당 조성을 확인한 결과 Man, Gal 및 GalA를 높은 비율로 함유하고 있었다. 본 당쇄의 결합양식을 규명하기 위하여 methlyation analysis를 행한 결과 CE-4a는 terminal Galp. 3-linked-Galp, 4-linked Manp, 2,4,6-linked Manp를 포함한 총 10종의 결합으로 구성되어 있었다. 또한 CE-4a의 전체구조를 추정하기 위하여 endo-α-(1→4)-polygalacturonase, exo-α-galactosidase 및 endo-β-(1→4)-mannanase를 이용한 연속 가수분해 처리 및 해석을 행하였다. 결과를 종합하면, 토란 유래 다당 CE-4a는 (1→4)-mannan 주쇄로 존재하며 주쇄인 mannose의 C(O)6 위치에서 한가닥 또는 C(O)2, C(O)6 위치에서 동시에 두가닥의 측쇄가 연결되어 존재하고, 측쇄는 주로 galacto oligo당이 측쇄로 분지된 특징이 있음을 확인할 수 있었다.

노로바이러스 및 로타바이러스 감염의 역학 및 기후요인과의 관계: 천안시, 2010-2019 (Molecular epidemiologic trends of norovirus and rotavirus infection and relation with climate factors: Cheonan, Korea, 2010-2019)

  • 오은주;김장묵;김재경
    • 디지털융복합연구
    • /
    • 제18권12호
    • /
    • pp.425-434
    • /
    • 2020
  • 바이러스감염으로 인한 설사는 전 세계적으로 공중 보건의 주요 문제이며 사망률의 큰 부분을 차지하고 있지만 기후데이터를 이용하여 분석한 연구는 많지 않다. 따라서 본 연구는 설사를 유발하는 바이러스인 Rotavirus Gr.A, Norovirus G-I & GII의 감염과 기후와의 인과관계를 분석하여 조기진단과 치료를 용이하게 하고 계절성 질환을 더욱 관리하고 예방하는데 기여하고자 하였다. 2010년 6월부터 2019년 12월까지 단국대학교 병원에서 시행한 대변 시료 4,009개의 설사바이러스 6종의 멀티플렉스 역전사 PCR(mRT-PCR)검사결과와 다양한 기후 요인 중 체감온도, 상대습도, 일조율과의 상관관계를 후향적으로 분석하였다. 4,009 개의 대변 샘플 중 985 개는 Rotavirus Gr.A, Norovirus G-I & GII 감염에 대해 양성이었다. 이 985 건 중 95.3 % (n = 939)는 10 세 미만으로 Rotavirus Gr.A, Norovirus G-I & GII는 10 세 미만 환자에서 높은 감염률을 보였다. 우리는 기상 빅데이터와 연령, 시기별 감염분석 등에 기초하여 Rotavirus Gr.A의 감염이 상대 습도에 따라 유의한 상관 관계가 있음을 확인하였고 Norovirus G-II의 감염은 체감 온도와 유의한 상관 관계가 있음을 확인하였다. 본 연구는 기후요인에 따른 Rotavirus Gr.A, Norovirus G-I & GII 감염의 분포에 대한 이해도를 향상시킴으로써 바이러스성 설사질환과 관련된 보건정책의 설정이나 계절적 영향에 대한 새로운 통찰력을 제공하는데 유용한 자료가 될 것으로 기대한다.

형질전환 초파리를 이용한 Mdr49A 유전자의 살충제 교차저항성 기능 구명 (Molecular Mechanism of ABC Transporter Mdr49A Associated with a Positive Cross-Resistance in Transgenic Drosophila)

  • 성건묵
    • 한국응용곤충학회지
    • /
    • 제59권4호
    • /
    • pp.341-348
    • /
    • 2020
  • ATP-binding cassette (ABC) transporter는 다양한 기질을 세포 밖과 세포 안으로 수송하는 대표적인 수송단백질이다. 곤충에서 ABC transporter는 살충제에 대한 저항성을 발달시키는 중요한 역할을 한다. 현재까지 모델곤충인 초파리를 대상으로 ABC transporter의 살충제 교차저항성에 관한 연구는 많이 수행되어오지 않았다. 본 연구에서는 ABC transporter에 속하는 Mdr49A 유전자가 여섯 종류의 살충제에 보이는 교차저항성 기작을 형질전환 초파리를 이용하여 구명하였다. 초파리 91-R과 91-C 계통은 공통된 조상으로부터 유래되었으며 91-R은 60년 이상 DDT에 노출되었지만 91-C는 어떠한 살충제에도 노출되지 않고 유지되어 왔다. 91-R 계통의 MDR49A 단백질에서 유래된 3개의 아미노산 돌연변이를 형질전환 초파리에 과발현 시켰을 때 carbofuran에 대해서 2.0~6.7배 그리고 permethrin에 대해서 2.5~10.5배의 교차저항성을 나타낸 반면 다른 약제, abamectin, imidacloprid, methoxychlor, prothiofos에 대해서는 어떠한 교차저항성도 나타내지 않았다. 이상의 결과는 Mdr49A 유전자의 과발현과 더불어 3개의 아미노산 돌연변이는 두 개 약제, carbofuran과 permethrin에 대해 교차저항성 기능을 한다고 제시하고 있다.

Alterations of mRNA and lncRNA profiles associated with the extracellular matrix and spermatogenesis in goats

  • Chen, Haolin;Miao Xiaomeng;Xu, Jinge;Pu, Ling;Li, Liang;Han, Yong;Mao, Fengxian;Ma, Youji
    • Animal Bioscience
    • /
    • 제35권4호
    • /
    • pp.544-555
    • /
    • 2022
  • Objective: Spermatozoa are produced within the seminiferous tubules after sexual maturity. The expression levels of mRNAs and lncRNAs in testicular tissues are different at each stage of testicular development and are closely related to formation of the extracellular matrix (ECM) and spermatogenesis. Therefore, we set out to study the expression of lncRNAs and mRNAs during the different developmental stages of the goat testis. Methods: We constructed 12 RNA libraries using testicular tissues from goats aged 3, 6, and 12 months, and studied the functions of mRNAs and lncRNAs using the gene ontogeny (GO) and Kyoto encyclopedia of genes and genomes (KEGG) databases. Relationships between differentially expressed genes (DEGs) were analyzed by lncRNA-mRNA co-expression network and protein-protein interaction network (PPI). Finally, the protein expression levels of matrix metalloproteinase 2 (MMP2), insulin-like growth factor 2 (IGF2), and insulin-like growth factor-binding protein 6 (IGFBP6) were detected by western blotting. Results: We found 23, 8, and 135 differentially expressed lncRNAs and 161, 12, and 665 differentially expressed mRNAs that were identified between 3 vs 6, 6 vs 12, and 3 vs 12 months, respectively. GO, KEGG, and PPI analyses showed that the differential genes were mainly related to the ECM. Moreover, MMP2 was a hub gene and co-expressed with the lncRNA TCONS-0002139 and TCONS-00093342. The results of quantitative reverse-transcription polymerase chain reaction verification were consistent with those of RNA-seq sequencing. The expression trends of MMP2, IGF2, and IGFBP6 protein were the same as that of mRNA, which all decreased with age. IGF2 and MMP2 were significantly different in the 3 vs 6-month-old group (p<0.05). Conclusion: These results improve our understanding of the molecular mechanisms involved in sexual maturation of the goat testis.

Anesthetic efficacy of supplemental buccal infiltration versus intraligamentary injection in mandibular first and second molars with irreversible pulpitis: a prospective randomized clinical trial

  • Zargar, Nazanin;Shojaeian, Shiva;Vatankhah, Mohammadreza;Heidaryan, Shirin;Ashraf, Hengameh;Baghban, Alireza Akbarzadeh;Dianat, Omid
    • Journal of Dental Anesthesia and Pain Medicine
    • /
    • 제22권5호
    • /
    • pp.339-348
    • /
    • 2022
  • Background: To compare the anesthetic efficacy of supplemental buccal infiltration (BI) (1.7 ml) versus intraligamentary (IL) injection containing 0.4 ml of 4% articaine with 1:100.000 epinephrine after an inferior alveolar nerve block (IANB) with 1.7 ml 2% lidocaine in the first and second mandibular molars diagnosed with irreversible pulpitis (IP). Methods: One hundred subjects diagnosed with IP of either the mandibular first (n = 50) or second molars (n = 50) and failed profound anesthesia following an IANB were selected. They randomly received either the IL or BI techniques of anesthesia. Pain scores on a 170 mm Heft-Parker visual analog scale were recorded initially, before, and during supplemental injections. Furthermore, pulse rate was measured before and after each supplemental injection. During the access cavity preparation and initial filing, no or mild pain was assumed to indicate anesthetic success. The chi-square test, Mann-Whitney U test, and independent samples t-test were used for the analyses. Results: The overall success rates were 80% in the IL group and 74% in the BI group, with no significant difference (P = 0.63). In the first molars, there was no significant difference between the two techniques (P = 0.088). In the second molars, IL injection resulted in a significantly higher success rate (P = 0.017) than BI. IL injection was statistically more successful (P = 0.034) in the second molars (92%) than in the first molars (68%). However, BI was significantly more successful (P = 0.047) in the first molars (88%) than in the second molars (64%). The mean pulse rate increase was significantly higher in the IL group than in the BI group (P < 0.001). Conclusions: Both the IL and BI techniques were advantageous when used as supplemental injections. However, more favorable outcomes were observed when the second molars received IL injection and the first molars received BI.