Effect of Different Ingredients in Traditional Korean Medicine for Human Uterine Leiomyoma on Normal Myometrial and Leiomyomal Smooth Muscle Cell Proliferation

Prati Bajracharya,이은주,이동목,Sang Hee Shim,김극준,Sung Ho Lee,Jei Jun Bae,전상식,Tae Kyun Lee,Seok Hoon Kwon,최인호 대한약학회 2009 Archives of Pharmacal Research Vol.32 No.11

Crude water extracts of 13 traditional Korean medicinal ingredients used for leiomyomal treatment were prepared and used to treat human uterine normal myometrial and leiomyomal cell cultures. All the ingredients inhibited proliferation and altered the morphology of both myometrial and leiomyomal cells. Among the 13 ingredients, n-hexane-, chloroform-, and ethylacetate-soluble fractions were extracted from seven ingredients that potently inhibited cell proliferation in their water extract form. Among these, the ethylacetate-fraction of Phlomis umbrosa and Spatholobus suberectus, and the chloroform-fraction of Curcuma zedoaria and S. suberectus inhibited leiomyomal cell proliferation significantly compared to myometrial cell proliferation. Similarly, immunohistochemical analysis showed the inhibition of transforming growth factor-beta receptor 2 in leiomyomal tissue after treatment with the fractions of the ingredients. Moreover, the chloroform-fraction of C. zedoaria was subfractionated by open column chromatography. Two of the eight subfractions (fractions 6 and 7) potently inhibited cell proliferation in leiomyoma compared to myometrium. Further study will be performed with the goal of isolating specific compounds from two effective subfractions of C. zedoaria, ethylacetate-fraction of P. umbrosa, and the ethylacetate and chloroform-fractions of S. suberectus. The present study may be helpful in developing an alternative remedy to leiomyoma with minimal side-effects compared to the current treatments.

Effect of microencapsulation of <i>Lactobacillus salivarus</i> 29 into alginate/chitosan/alginate microcapsules on viability and cytokine induction

Bajracharya, Prati,Islam, Mohammad Ariful,Jiang, Tao,Kang, Sang-Kee,Choi, Yun-Jaie,Cho, Chong-Su Informa UK, Ltd. 2012 Journal of microencapsulation Vol.29 No.5

<P>Harsh gastric condition causes low bioavailability of probiotics when supplied orally. Polymeric encapsulation has successfully protected bacteria from harsh gastric condition and ultimately increased persistency and multiplication at the targeted region. In this study, we encapsulated LS29 into ACA microcapsules and characterized them. The survivability and release of LS29 from LS29-loaded ACA microcapsules in SGF and SIF were studied. Encapsulation efficiency of LS29 in ACA microcapsules was 99.9%. Approximately 70% of bacteria survived at pH 2 by 120 min after encapsulation. Although not much difference of the survivability of LS29 encapsulated into ACA and FDACA was observed, freeze-drying improved the controlled-release of LS29 in SIF and also showed better storage survivability at 4°C for 8 weeks. Furthermore, investigation of <I>in vitro</I> production of cytokines in RAW264.7 showed high level of induction of TNF-&agr; and IL-10. These <I>in vitro</I> results support that the LS29 might have a balanced immunomodulatory effect.</P>

Gene expression profiles analyzed by DNA sequencing of cDNA clones constructed from porcine preadipocytes and adipocytes

이동목,Prati Bajracharya,장은정,이은주,채성화,박홍석,강세원,이용석,이기호,장종수,정정수,최인호 한국유전학회 2012 Genes & Genomics Vol.34 No.2

As a step towards understanding the molecular mechanism of adipogenesis in pigs, preadipocytes purified from the back fat of 1 day-old female piglets were used for in vitro culture. Normalized cDNA libraries were constructed with 1.6×107 and 1.1×107 independent clones from preadipocyte and mature adipocyte mRNAs, respectively. Polymerase chain reaction (PCR)result using primers T3 and T7 (universal primer) confirmed the presence of the insert in the vector. Sequencing of 2,112randomly selected clones from each cDNA library identified 217 clusters, 1,169 singletons, and 216 contigs in preadipocytes and 231 clusters, 1,100 singletons, and 233 contigs in mature adipocytes. Expressed sequence tag (EST) identified 24 genes with known annotation highly expressed in adipocytes and 21 in preadipocytes by at least four EST number. Among those 45 genes, when analyzed by real time RT-PCR,76% of the gene showed significant difference between preadipocytes and mature adipocytes. Highly expressed genes in mature adipocytes were related to adipogenesis, extracellular matrix control and oncogenes, whereas cytoskeleton-related genes were down-regulated. An interesting similarity found during gene profile studies indicated a correlation between cancer and adipogenesis.

Gene expression profiles during differentiation and transdifferentiation of bovine myogenic satellite cells

이은주,Prati Bajracharya,이동목,강세원,이용석,이현정,홍성구,장종수,김재우,Robert D. Schnabel,Jeremy F. Taylor,최인호 한국유전학회 2012 Genes & Genomics Vol.34 No.2

The molecular mechanisms underlying myogenic satellite cells (MSCs) differentiation into myotube-formed cells (MFCs) and transdifferentiation into adipocyte-like cells (ALCs) are unclear. As a step towards understanding the molecular mechanisms underlying MSC differentiation and transdifferentiation,we attempted to identify the genes differentially expressed during differentiation and transdifferentiation using gene microarray analysis (GMA). Thirty oligonucleotide arrays were used with two technical replicates and nine and six biological replicates for MFCs vs. MSCs and ALCs vs. MSCs, respectively,to contrast expression profile differences. GMA identified 1,224 differentially expressed genes by at least 2-fold during differentiation and transdifferentiation of MSCs. To select the highly expressed genes for future functional study, genes with a 4-fold expression difference were selected for validation by real time RT-PCR and approximately 96.9% of the genes were validated. The up-regulation of marker genes for myogenesis (MYL2, MYH3) and adipogenesis (PPARγ, and FABP4) was observed during the differentiation and transdifferentiation of MSCs into MFCs and ALCs, respectively. KOG analysis revealed that the most of the genes up-regulated during differentiation and transdifferentiation of MSCs were related to signal transduction. Again the exact location of 109 differentially expressed genes by 4-fold were analyzed by chromosome mapping. Among those, co-localization of 29 genes up-regulated during transdifferentiation with QTL for marbling score and intramuscular fat percentage supports the involvement of these genes in cellular transdifferentiation. Interestingly, some genes with unknown function were also identified during the process. Functional studies on these genes may unfold the molecular mechanisms controlling MSC differentiation and transdifferentiation.

Gene Expression Profiles of Uterine Normal Myometrium and Leiomyoma and Their Estrogen Responsiveness In Vitro

최인호,이은주,Prati Bajracharya,이동목,조경현,김극준,배영경,김미진,이기호,김항진,송건호,전상식 대한병리학회 2010 Journal of Pathology and Translational Medicine Vol.44 No.3

Background : Uterine leiomyomas are common benign smooth muscle tumors among the reproductive aged-women. The research has been aimed to identify the differentially expressed genes between normal myometrium and leiomyoma and to investigate the effects of E2 on their expression. Methods : Gene microarray analysis was performed to identify the differentially expressed genes between normal myomerium and leiomyoma. The data was confirmed at protein level by tissue microarray. Results : Gene microarray analysis revealed 792 upregulated genes in leiomyoma. Four genes (tropomyosin 4 [TPM4], collagen, type IV, alpha 2 [COL4α2], insulin-like growth factor binding protein 5 [IGFBP5], tripartite motif-containing 28 [TRIM28]) showed the most dramatic upregulation in all leiomyoma samples. Tissue microarray analyses of 262 sample pairs showed significantly elevated expression of TPM4, IGFBP5, estrogen receptor-α, and progesterone receptor (PR) protein in leiomyoma from the patients in theirforties, COL4α2 in the forties and fifties age-groups, and TRIM28 in the thirties age-group. PR, insulin-like growth factor 1 (IGF-1), IGF-1 receptor (IGF-1R) and IGFBP5 were induced by E2 in in vitro culture of tissue explants from which cells migrated throughout the plate. Among these, PR, IGF-1, IGFBP5 genes showed higher expression in tissue compared to cells-derived from tissue in leiomyoma and IGF-1R in leiomyoma cell. Conclusions : This observation implies the importance of the whole tissue context including the cells-derived from tissue in the research for the understanding of molecular mechanism of leiomyoma. Here, we report higher expression of TRIM28 in leiomyoma for the first time and identify E2-responsive genes that may have important roles in leiomyoma development. Background : Uterine leiomyomas are common benign smooth muscle tumors among the reproductive aged-women. The research has been aimed to identify the differentially expressed genes between normal myometrium and leiomyoma and to investigate the effects of E2 on their expression. Methods : Gene microarray analysis was performed to identify the differentially expressed genes between normal myomerium and leiomyoma. The data was confirmed at protein level by tissue microarray. Results : Gene microarray analysis revealed 792 upregulated genes in leiomyoma. Four genes (tropomyosin 4 [TPM4], collagen, type IV, alpha 2 [COL4α2], insulin-like growth factor binding protein 5 [IGFBP5], tripartite motif-containing 28 [TRIM28]) showed the most dramatic upregulation in all leiomyoma samples. Tissue microarray analyses of 262 sample pairs showed significantly elevated expression of TPM4, IGFBP5, estrogen receptor-α, and progesterone receptor (PR) protein in leiomyoma from the patients in theirforties, COL4α2 in the forties and fifties age-groups, and TRIM28 in the thirties age-group. PR, insulin-like growth factor 1 (IGF-1), IGF-1 receptor (IGF-1R) and IGFBP5 were induced by E2 in in vitro culture of tissue explants from which cells migrated throughout the plate. Among these, PR, IGF-1, IGFBP5 genes showed higher expression in tissue compared to cells-derived from tissue in leiomyoma and IGF-1R in leiomyoma cell. Conclusions : This observation implies the importance of the whole tissue context including the cells-derived from tissue in the research for the understanding of molecular mechanism of leiomyoma. Here, we report higher expression of TRIM28 in leiomyoma for the first time and identify E2-responsive genes that may have important roles in leiomyoma development.

Oxidative Stabilization of Conjugated Linoleic Acid by One-Pot PEGylation

문현석,조종수,서지혜,Ding-Ding Guo,이홍구,Prati Bajracharya,최윤재,정근수,박종래 한국고분자학회 2011 Macromolecular Research Vol.19 No.8

This study describes a novel and one-step procedure for the PEGylation of conjugated linoleic acid (CLA)in the melting state without solvents or catalysts for oxidative stabilization of CLA. The double bonds of CLA in PEGylated CLA (PCLA) were clearly detected by 1H NMR (nuclear magnetic resonance) spectroscopy, indicating the successful PEGylation of CLA without oxidation during PEGylation, even at high temperatures. The level of headspace oxygen depletion by PCLA was lower than CLA, indicating that the oxidation of CLA was protected by PEGylation. This easy and simple one-pot PEGylation of CLA is an effective way of preparing fatty acids with increased stability to oxidation, which would be quite useful for nutritional and pharmaceutical studies.

Production and Regulation of Nandrolone in Porcine Leydig Cell Culture

Dong Mok Lee,Eun Ju Lee,Prati Bajracharya,Inho Choi 한국동물생명공학회(구 한국동물번식학회) 2008 Reproductive & developmental biology Vol.32 No.2

Production and Regulation of Nandrolone in Porcine Leydig Cell Culture

Dong Mok Lee,Eun Ju Lee,Prati Bajracharya,Inho Choi 한국동물번식학회 2008 Reproductive & Developmental Biology(Supplement) Vol.32 No.2s

cDNA Microarray Analysis of Differential Gene Expression in Boar Testes during the Prepubertal Period

Lee, Dong-Mok,Lee, Ki-Ho,Choi, Jin Ho,Hyun, Jin Hee,Lee, Eun Ju,Bajracharya, Prati,Lee, Yong Seok,Chang, Jongsoo,Chung, Chung Soo,Choi, Inho Asian Australasian Association of Animal Productio 2009 Animal Bioscience Vol.22 No.8

In an attempt to understand the biochemical mechanism for the synthesis of the anabolic steroid, 19-nortestosterone, produced by prepubertal boar testes and its physiological role, normalized complementary DNA (cDNA) from boar testes was generated. DNA sequencing of 2,016 randomly selected clones yielded 794,116 base pairs of high quality nucleotide sequence. Computer-assisted assembly of the nucleotide sequence of each clone resulted in 423 contigs and 403 singletons including several genes for steroidogenic enzymes and molecules related to steroid metabolism. Analysis of gene expression pattern by use of the presently-fabricated cDNA microarray identified a number of genes that were differentially expressed during the postnatal development period in boar testes. Two genes of unknown function were identified to be highly expressed in the testis of 2-weeks-old neonatal boar. In addition, the sequencing of open reading frames of these genes revealed their homology with human alpha hemoglobin and Homo sapiens hypothetical LOC643669, transcript variant 1. Moreover, the transcripts of these genes were also detected in porcine muscle and adipocytes, in addition to Leydig cells of pigs.