The classical and a non-classical pathways associated with NF-κB are involved in estrogen-medicated regulation of Calbindin-D9k gene in rat pituitary cells

Lee, G.S.,Choi, K.C.,Han, H.J.,Jeung, E.B. North-Holland 2007 Molecular and cellular endocrinology Vol.277 No.1-2

Calbindin-D9k (CaBP-9k) is a high affinity calcium binding protein that is highly expressed in the duodenum, kidney, uterus, and pituitary glands. Previous studies have shown that CaBP-9k expression is regulated by several steroid hormones, such as 1,25-dihydroxyvitamin D3, glucocorticoids, 17β-estradiol (E2), and progesterone (P4), in a tissue-specific manner. However, the promoter elements that mediate transcriptional regulation by these steroid hormones are not clearly understood, mainly due to the lack of an appropriate cell line expressing CaBP-9k. Recently it was shown that CaBP-9k was constitutively expressed in the rat pituitary gland, and is expressed in an E2-dependent manner in a pituitary gland tumor-derived cell line, GH3. In the current study, we examined the activity of the estrogen responsive element (ERE) in rat CaBP-9k gene in GH3 cells, using a luciferase gene reporter assay, electrophoretic mobility shift assay, and mutagenesis. A nuclear factor-κB (NF-κB) binding site in the CaBP-9k promoter region was identified (nucleotides -848 to -834 from the transcriptional start site), and we demonstrated that addition of an NF-κB blocker to GH3 cells reduced E2-induced CaBP-9k transcription. In the present study, we further showed a previously reported imperfect ERE (nucleotides +51 to +65) between exon I and intron A of CaBP-9k, indicating that the interaction of estrogen receptor (ER) α with this region is involved in the regulation of CaBP-9k promoter activity and its expression. Taken together, these results suggest that in GH3 cells, both the classical ERα-ERE pathway and a non-classical pathway involving NF-κB are involved in E2-mediatd regulation of CaBP-9k expression in the pituitary gland.

Proposed mechanism in the change of cellular composition in the outer medullary collecting duct during potassium homeostasis.

Park, E-Y,Kim, W-Y,Kim, Y-M,Lee, J-H,Han, K-H,Weiner, I D,Kim, J Gutenberg 2012 HISTOLOGY AND HISTOPATHOLOGY Vol.27 No.12

<P>Potassium depletion (K?-D) induces hypertrophy and hyperplasia of collecting duct cells, and potassium repletion (K?-R) induces regression of these changes. The purpose of this study was to examine the time courses of the changes in cellular composition, the origin of intercalated cells (ICs) and the mechanism responsible for these changes. SD rats received K?-depleted diets for 1, 7, or 14 days. After K?-D for 14 days some of the rats received normal diets for 1, 3, 5, or 7 days. In the inner stripe of the outer medulla, K?-D increased significantly the number and proportion of H?-ATPase-positive ICs, but decreased the proportion of H?-ATPase-negative principal cells (PCs). However, proliferation was limited to H?-ATPase-negative PCs. During K?-R, the cellular composition was recovered to control level. Apoptosis increased during K?-R and exclusively limited in H?-ATPase-negative PCs. Double immunolabeling with antibodies to PC and IC markers identified both cells negative or positive for all markers during both K?-D and K?-R. Electron microscopic observation showed that ultrastructure of AE1-positive some cells were similar to AE1-negative some cells during K?-R. LC3 protein expression increased significantly and autophagic vacuoles appeared particularly in PCs on days 14 of K?-D and in ICs on days 3 of K?-R. These results suggest that PCs and ICs may interconvert in response to changes in dietary K+ availability and that autophagic pathways may be involved in the interconversion.</P>

소 모색관련 유전자 MC1R 의 RCR - RFLP Marker 를 이용한 한우육 판별

정의룡,김우태,김연수,한상기 한국동물자원과학회 2000 한국축산학회지 Vol.42 No.4

The melanocortin 1 receptor(MClR) plays a central role in regulation of eumelanin(black/brown) and phaeomelanin(red/yellow) pigment synthesis within the mammalian melanocytes. Mutations within the gene encoding MC1R have been shown to explain coat colour variations within several species including cattle. This study was performed to develop the identification technique of Hanwoo meat using MC1R gene associated with the coat colors of cattle. Alleles of the MC1R locus were detected by PCR-RFLP analysis and genotype frequency and DNA sequences of MC1R gene were compared among cattle breeds. Genomic DNA was extracted from meat or blood samples of five breeds including Hanwoo(n=200), Holstein(n=100), Angus(n=20), Hereford(n=20) and Charolais(n=20). The MC1R gene was used to amplify 739bp and 173bp of the bovine E-locus corresponding to positions 228-966 and 318-490, respectively, using the two specific primers. The amplified products were digested with Bse118 I or Msp I and Aci I enzymes, and DNA fragments were separated by gel electrophoresis for RFLP genotype analysis. Six genotypes, E^D/E^D E^D/E^+, E^D/e, E^+/E^+,E^+/e and e/e, controlled by three alleles E^D, E^+ and e were observed in MC1 locus. When the amplified DNA product(739bp) was digested with Bse118 I enzyme, Hanwoo meat showed a single band of 739bp, whereas two fragments of 531bp and 208bp were detected in Holstein meat and Angus breed, respectively. Also, in the RFLP patterns using Msp I enzyme, Hanwoo meat produced two fragments of 535bp and 174bp, while three fragments of 328bp, 207bp and 174bp were observed in Holstein meat and Angus breeds, respectively. Therefore, breed-specific RFLP markers showing distinct differences between these breeds were found by PCR-RFLP analysis. When the amplified DNA product(173bp) was digested with Aci I enzyme to classify subtype of E allele, the E^D allele produced three fragments of 97, 68 and 8bp, while the E^+ and d alleles produced two fragments of 173 and 8bp according to the Aci I recognition sequence. Among the six genotypes, two genotypes of E^+/e and e/e were observed in Hanwoo and their frequencies were 0.07 and 0.93, respectively. However, the E^D/ED and E^D/e genotypes were present in Holstein and E^D/E^D, E^D/E^+ and E^D/e genotypes in Angus breeds. Therefore, the E^+/e and e/e genotypes observed in Hanwoo and E^D/E^D, E^D/E^+ and E^D/e genotypes detected only in Holstein and Angus breeds may be useful as breed-specific DNA markers for distinguishing between Hanwoo meat and Holstein and Angus meats. When comparing MC1R sequences among Hanwoo, Holstein and Angus, a Gly → Val amino acid change due to a single base(G) deletion at colon 104 was found in Hanwoo. Consequently, breed specific RFLP genotypes of MC1R gene related to bovine coat colors could be used as DNA markers for identification of Hanwoo meat from Holstein and Angus meats.

PCR - RFLP 기법을 이용해 젖소개량을 위한 유전적 표지로서 K- Casein 좌위의 유전자형 분석

정의룡(E . R . Chung),김우태(W . T . Kim),최석호(S . H . Choi),임태진(T . J . Rhim),한상기(S . K . Han) 한국축산학회 1995 한국축산학회지 Vol.37 No.1

Genotypes of K-casein(K-CN) locus as a genetic marker linked to quantitative trait loci affecting traits of economic importance in dairy cattle were determined by PCR-RFLP method. Genomic DNA was prepared from blood of Holstein cows. The PCR was used to amplify an 874 by region between nucleotides 10592 and 11466 from exon IV to intron IV of the bovine K-CN gene using sense primer(5`-GTGCTGAGTAGGTATCCTAG-3`) and antisense primer(5`GTAGAGTGCAACAACACTGG-3`). After amplification, PCR products were digested with four restriction enzymes, Hind III, Rsa I, Taq I, and Pst I, and the fragments were separated by agarose gel electrophoresis for RFLP analysis of K-CN locus. In addition to screening for the known Hind III and Rsa I restriction site polymorphisms of K-CN locus, we have found additional RFLPs specific for the K-CN A and B alleles in Taca I and Pst I enzymes. The amplified DNA product digested with each restriction enzyme generated specific RFLP pattern that allowed precise identification of K-CN AA, BB or AB genotypes. The K-CN genotypes determined for cows by the PCR-RFLP method agreed completely with the phenotypes obtained from milk samples of the same individuals. Thus, PCR amplification and RFLP analysis was shown to be a rapid and sensitive method for the discrimination of K-CN genotypes directly at the DNA level in dairy cattle of any age or sex. Consequently, the PCR-RFLP method presented in this study can be used as a valuable tool for early selection of AI bulls and calves with desirable K-CN B gene or K-CN BB genotype affecting superior milk production traits for genetic improvement of Holstein dairy cattle.

Study of B^+/-→K^+/-(K_SKπ)⁰ decay and determination of η_c and η_c(2S) parameters

Belle Collaboration,Vinokurova, A.,Kuzmin, A.,Eidelman, S.,Arinstein, K.,Aulchenko, V.,Aushev, T.,Bakich, A.M.,Balagura, V.,Barberio, E.,Belous, K.,Bhardwaj, V.,Bondar, A.,Bozek, A.,Bracko, M.,Brodzic North-Holland Pub. Co 2011 Physics letters: B Vol.706 No.2

We report the results of a study of B<SUP>+/-</SUP>→K<SUP>+/-</SUP>η<SUB>c</SUB> and B<SUP>+/-</SUP>→K<SUP>+/-</SUP>η<SUB>c</SUB>(2S) decays followed by η<SUB>c</SUB> and η<SUB>c</SUB>(2S) decays to (K<SUB>S</SUB>Kπ)<SUP>0</SUP>. The results are obtained from a data sample containing 535 million BB@?-meson pairs collected by the Belle experiment at the KEKB e<SUP>+</SUP>e<SUP>-</SUP> collider. We measure the products of the branching fractions B(B<SUP>+/-</SUP>→K<SUP>+/-</SUP>η<SUB>c</SUB>)B(η<SUB>c</SUB>→K<SUB>S</SUB>K<SUP>+/-</SUP>π<SUP>@?</SUP>)=(26.7+/-1.4(stat)<SUB>-2.6</SUB><SUP>+2.9</SUP>(syst)+/-4.9(model))x10<SUP>-6</SUP> and B(B<SUP>+/-</SUP>→K<SUP>+/-</SUP>η<SUB>c</SUB>(2S))B(η<SUB>c</SUB>(2S)→K<SUB>S</SUB>K<SUP>+/-</SUP>π<SUP>@?</SUP>)=(3.4<SUB>-1.5</SUB><SUP>+2.2</SUP>(stat+model)<SUB>-0.4</SUB><SUP>+0.5</SUP>(syst))x10<SUP>-6</SUP>. Interference with the non-resonant component leads to significant model uncertainty in the measurement of these product branching fractions. Our analysis accounts for this interference and allows the model uncertainty to be reduced. We also obtain the following charmonia masses and widths: M(η<SUB>c</SUB>)=(2985.4+/-1.5(stat)<SUB>-2.0</SUB><SUP>+0.5</SUP>(syst)) MeV/c<SUP>2</SUP>, Γ(η<SUB>c</SUB>)=(35.1+/-3.1(stat)<SUB>-1.6</SUB><SUP>+1.0</SUP>(syst)) MeV/c<SUP>2</SUP>, M(η<SUB>c</SUB>(2S))=(3636.1<SUB>-4.2</SUB><SUP>+3.9</SUP>(stat+model)<SUB>-2.0</SUB><SUP>+0.7</SUP>(syst)) MeV/c<SUP>2</SUP>, Γ(η<SUB>c</SUB>(2S))=(6.6<SUB>-5.1</SUB><SUP>+8.4</SUP>(stat+model)<SUB>-0.9</SUB><SUP>+2.6</SUP>(syst)) MeV/c<SUP>2</SUP>.

Aerosol delivery of eukaryotic translation initiation factor 4E-binding protein 1 effectively suppresses lung tumorigenesis in K-ras^LA1 mice

Chang, S-H,Kim, J-E,Lee, J-H,Minai-Tehrani, A,Han, K,Chae, C,Cho, Y-H,Yun, J-H,Park, K,Kim, Y-S,Cho, M-H Nature America, Inc. 2013 Cancer gene therapy Vol.20 No.6

Conventional radiotherapy or chemotherapy for the long-term survival of patients with lung cancer is still difficult for treatment in metastatic and advanced tumors. Therefore, the safe and effective approaches to the treatment of lung cancer are needed. In this study, the effect of delivered eukaryotic translation initiation factor 4E (eIF4E)-binding protein 1 (4E-BP1) on lung cancer progression was evaluated. Recombinant adeno-associated virus (rAAV)-M3/4E-BP1 was delivered into 6-week-old K-ras<SUP>LA1</SUP> lung cancer model mice through a nose-only inhalation system twice a week for 4 weeks. Long-term repeated delivery of 4E-BP1 effectively reduced tumor progression in the lungs of K-ras<SUP>LA1</SUP> mice. Reduction of eIF4E by overexpression of 4E-BP1 resulted in suppression of cap-dependent protein expression of basic fibroblast growth factor (bFGF or FGF-2) and vascular endothelial growth factor (VEGF). In addition, delivered 4E-BP1 inhibited the proliferation of lung cancer cells in K-ras<SUP>LA1</SUP> mice model. Our results suggest that long-term repeated viral delivery of 4E-BP1 may provide a useful tool for designing lung cancer treatment.

Bacteroides fragilis의 독소유전자 분석 및 재조합독소 발현에 관한 연구

오희복,정경태,이기은,성원근,이용진,이근영 국립보건연구원 1999 국립보건원보 Vol.36 No.-

Partition energy of complete product of circulant graphs and some new class of graphs

E. Sampathkumar,S. V. Roopa,K. A. Vidya,M. A. Sriraj 장전수학회 2018 Advanced Studies in Contemporary Mathematics Vol.28 No.2

Let G = (V,E) be a graph and Pk = {V1, V2, ..., Vk} be a partition of V . The L-matrix with respect to a partition Pk of the vertex set V of graph G of order n is the unique square symmetric matrix Pk(G) = [aij ] with zero diagonal, whose entries aij with i 6≠ j are defined as follows: (i) If vi, vj ∈ Vr, then aij = 2 or −1 according as vivj is an edge or not. (ii) If vi ∈ Vr and vj ∈ Vs for r 6≠s, then aij = 1 or 0 according as vivj is an edge or not. For all Vi and Vj in Pk, i 6≠j remove the edges between vertices of Vi and Vj and add the edges between the vertices of Vi and Vj which are not in G, the resulting graph is called k-complement of G and is denoted by (G)k. For each set Vr in Pk, remove the edges of G joining the vertices within Vr and add the edges of G (complement of G) joining the vertices of Vr, the graph obtained is called k(i)-complement and is denoted by (G)k(i). The k-partition energy of a graph G with respect to partition Pk is denoted by EPk (G) and is defined as the sum of the absolute values of k-partition eigenvalues of Pk(G). In this paper we construct some graphs such that the graph and its 2-complement are equienergetic with respect to a given partition. We also determine partition energy of complete product of m copies of a circulant graph G and its subgraph, their k-complement and k(i)-complement.

A study of γγ->K_S⁰K_S⁰ production at energies from 2.4 to 4.0 GeV at Belle

Belle Collaboration,Chen, W.T.,Abe, K.,Abe, K.,Adachi, I.,Aihara, H.,Anipko, D.,Aulchenko, V.,Bakich, A.M.,Barberio, E.,Bay, A.,Bedny, I.,Bitenc, U.,Bizjak, I.,Blyth, S.,Bondar, A.,Bozek, A.,Bracko, M North-Holland Pub. Co 2007 Physics letters: B Vol.651 No.1

K<SUB>S</SUB><SUP>0</SUP>K<SUB>S</SUB><SUP>0</SUP> production in two-photon collisions has been studied using a 397.6 fb<SUP>-1</SUP> data sample collected with the Belle detector at the KEKB e<SUP>+</SUP>e<SUP>-</SUP> collider. For the first time the cross sections are measured in the two-photon center-of-mass energy range between 2.4 GeV and 4.0 GeV and angular range |cosθ<SUP>*</SUP>|<0.6. Combining the results with measurements of γγ->K<SUP>+</SUP>K<SUP>-</SUP> from Belle, we observe that the cross section ratio σ(K<SUB>S</SUB><SUP>0</SUP>K<SUB>S</SUB><SUP>0</SUP>)/σ(K<SUP>+</SUP>K<SUP>-</SUP>) decreases from ∼0.13 to ∼0.01 with increasing energy. Signals for the χ<SUB>c0</SUB> and χ<SUB>c2</SUB> charmonium states are also observed.

PCR - SSCP 기법을 이용한 소 MC1R 유전자의 다형성 분석 및 한우육 감별

정의룡,김우태,김연수,한상기 한국동물자원과학회 2001 한국축산학회지 Vol.43 No.1

The single-strand conformation polymorphism(SSCP) is a point mutation screening method based on changes in the secondary structure in a defined single-stranded DNA fragment caused by a change in sequence. This study was performed to develop the identification technique of Hanwoo meat using PCR-SSCP marker of MC1R gene associated with the coat colors of cattle. Hanwoo, Holstein and imported cattle breeds(Angus, Hereford and Charolais) were used for the detection of PCR-SSCP genotypes of MC1R gene. The specific primers were used to amplify a 138bp and a 390bp of the bovine E-locus(including point mutation) corresponding to positions 318-455 and 318-707, respectively, in the MC1R gene. For SSCP genotype analysis, the PCR products were denatured at 95℃ for 5min, loaded on a 12-15% nondenaturing polyacrylamide gel and detected by silver staining. When the PCR product of 138bp was separated by gel electrophoresis, two genotypes of E^+/e and e/e were detected only in Hanwoo meat, but E^D/E^D and E^D/e and E^D/E^D genotypes were observed in Holstein meat and Angus breed, respectively. Also, in the SSCP analysis using PCR product of 390bp, Hanwoo meat produced banding pattern of two fragments, while single fragment was observed in Holstein meat and Angus breed. Therefore, breed-specific SSCP markers showing distinct differences between Hanwoo and Holstein and Angus were found by PCR-SSCP analysis. This study shows that the PCR-SSCP technique is relatively simple, fast and a practical tool for determination of MC1R genotypes of cattle breeds when compared with PCR-RFLP method. Therefore, the PCR-SSCP markers of MC1R gene could be used as DNA markers for identification of Hanwoo meat from Holstein and Angus meats.