• Title/Summary/Keyword: Human ovary

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Cloning and Nucleotide Sequence Analysis of the Virulence Gene Cassette from Vibrio cholerae KNIH002 Isolated in Korea (국내 분리주인 Vibrio cholerae KNIH002로부터 독성 유전자 카세트의 클로닝 및 염기서열 분석)

  • 신희정;박용춘;김영창
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.205-210
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    • 1999
  • 16brio cholerne is an important pathogenic organism that causes dimhea in human beings. V ciaoleroe KNIH002 was isolated from patients suffering with dian.heal disease in Korea. From Southern hybridization using the amplified PCR product of 307 bp as a probe. which was obtained from PCR reaction using primer detecting cholera toxin gene, we have found that the c b gene located in 4.5-kb fragmenl double digested with Pstl and BgllI of the chromosome. Therefore, we made mini-libraries of the isolate using PstI and Bgm restriction endonuclease and pBluescript SKU(+) vector. As a result. we cloned 4.5-kb PstI-BglII fragment containing the c a gene encoding a cholera toxin from the constructed mini-libraries of V olzolerae KNlH002 by colony hybridization using the same probes. This recombinant plasmid was named pCTX75. E. coii XL1- Blue harboring pCTX75 showed the cytotoxicity on Chinese Hamster Ovary cells. From the sequencing of he cloned recombinant plasmid, we confinned that it has virulence gene cassette consisting of ace, zot, ctx.4 and cf"~B gene. The ace and zot genes were composed of 291 hp and 1.200 bp with ATG initiation codon and TGA lennination codon, respectively. Nucleotide sequence of the ace gene exhibited 100% identity with that of V cholera E7946 El Tor Ogawa strains. But, nucleolide and amino acid sequence comparison of the zot gene exhibited 99% and 98.8% identity with that of V cholerae 395 Classical Ogawa stram, respectively. Specially. the Ala-100, Ala-272 and Ala-281 sites of Zoi polypeptide presented in V choleme 395 Classical Ogawa strain are replaced by Val in V cholerae KNIH002.

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Cloning of Pig Kidney cDNA Encoding an Angiotensin I Converting Enzyme (돼지 신장의 Angiotensin I Converting Enzyme cDNA 클로닝)

  • Yoon, Jang-Ho;Yoon, Joo-Ok;Hong, Kwang-Won
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.293-297
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    • 2006
  • Angiotensin converting enzyme(ACE) is a zinc-containing dipeptidase widely distributed in mammalian tissues and is thought to play a significant role in blood pressure regulation by hydrolyzing angiotensin I to the potent vasoconstrictor, angiotensin II. Recently, the presence of ACE in pig ovary was reported and the ACE from pig kidney was isolated and characterized. However no nucleotide sequence of the ACE gene from pig is yet known. We report here the cloning of the ACE cDNA from pig kidney by using the reverse transcriptase-polymerase chain reaction. The complete amino acid sequence deduced from the cDNA contains 1309 residues with a molecular mass of 150 kDa, beginning with a signal peptide of 33 amino acids. Amino acid sequence analysis showed that pig kidney ACE is also probably anchored by a short transmembrane domain located near the C-terminus. This protein contains a tandem duplication of the two homologous amino acid peptidase domain. Each of these two domains bears a putative metal-binding site (His-Glu-Met-Gly-His) identified in mammalian somatic ACE. The alignment of pig ACE amino acid sequence with human, rabbit, and mouse reveals that both two domains have been highly conserved during evolution.

STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.9-22
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    • 1974
  • Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

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Immunohistochemical study on the atretic and the growing follicles after experimental superovulation in rats 2. Atresia and growing of follicles (과배란 유기된 rat 난소에 퇴축난포와 성장난포에 대한 면역조직화학적 연구 2. 동원된 난포의 퇴축과 성장에 대하여)

  • Kwak, Soo-dong;Koh, Phil-ok;Kim, Chong-sup
    • Korean Journal of Veterinary Research
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    • v.37 no.1
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    • pp.79-86
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    • 1997
  • This study was designed to investigate the effects of superovulation on the growing and mature follicles following gonadotrophin treatments in mature rat by immunohistochemical methods. Eighteen mature rats (Sprague-Duwely, 190~230gm) were randomly alloted into 3 groups. One group was control group, another FSH-treated group was injected intramuscularly with 0.5 units of follicular stimulating hormone(FSH) / rat, and third PMS and HCG-treated group was injected intramuscularly with 20~25IU of pregnant mare serum(PMS) / rat and then at the 48 hrs later, with 20~25IU of human chorionic gonadotropin(HCG) / rat. Half the number of rats were administrated intraperitoneally with bromodeoxyuridine(Brdur, 0.2mg/gm BW once) at 2 hours before exanguination and the remainder of rats were sacrified without Brdur administration. The investigation by immunohistochemical methods using paraffin sections of ovaries was performed by using anti-Brdur antibody and PCNA(proliferating cell nuclear antigen) antibody for labeling proliferating cells in follicles. In immunohistochemical findings, follicles squeezed by peripheral corpus luteum or follicles large follicles with loosly and irregularly distributed granulosa cells and although with compacted granulosa cells, middle follicles with dilated round or oval follicular antrum were confirmed as atretic follicles. The proportions of atretic follicles in control group were 29.8%, 21.7% and 14.2% respectivley at large, middle and small follicles and mean proportions of these all 3 grade follicles were 26.7%. The proportions of atretic follicles in FSH-treated group were 35.4%, 24.9% and 10.4% respectively at large, middle and small follicles and mean proportions of these all 3 grade follicles were 28.1%. The proportions of atretic follicles in PMS and HCG-treated group were 44.7%, 24.0% and 12.7% respectively at large, middle and small follicles, and mean proportions of these all 3 grade follicles were 29.7%. The above findings reveal that the group with higher proportion of atretic follicles were ordered as large, middle and small follicles in size, and these proportions were increased in hormone treated two groups with more number of more growing and mature follicles when compared with control group.

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Partial Sequencing and Characterization of Porcine DNA Methyltransferase I cDNA

  • Lee, Y.Y.;Kim, M.S.;Park, J.J.;H.Y. Kang;Y.M. Chang;Yoon, J.T.;K.S. Min
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.84-84
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    • 2003
  • DNA methylation is involved in epigenetic processes such as X-chromosome inactivation, imprinting and silencing of transposons. DNA methylation is a highly plastic and critical component of mammalian development The DNA methyltransferases (Dnmts) are responsible for the generation of genomic methylation patterns, which lead to transcriptional silencing. The maintenance DNA methyltransferase enzyme, Dnmt 1, and the de novo methyltransferase, Dnmt3a and Dnmt3b, are indispensable for development because mice homozygous for the targeted disruption of any of these genes are not viable. The occurrence of DNA methylation is not random, and it can result in gene silencing The mechanisms underlying these processes are poorly understood. It is well established that DNA methylation and histone deacetylation operate along a common mechanistic pathway to repress transcription through the action of methyl-binding domain proteins (MBDs), which are components of, or recruit, histone deacetylase (HDAC) complexes to methylated DNA. As a basis for future studies on the role of the DNA-methyl-transferase in porcine development, we have isolated and characterized a partial cDNA coding for the porcine Dnmt1. Total RNA of testis, lung and ovary was isolated with TRlzol according to the manufacture's specifications. 5 ug of total RNA was reverse transcribed with Super Script II in the presence of porcine Dnmt 1 specific primers. Standard PCRs were performed in a total volume of 50 ul with cDNA as template. Two DNA fragmenets in different position were produced about 700bp, 1500bp and were cloned into pCR II-TOPO according to the manufacture's specification. Assembly of all sequences resulted in a cDNA from 158bp of 5'to 4861bp of 3'compare with the known human maintenance methyltransferase. Now, we are cloning the unknown Dnmt 1 region by 5'-RACE method and expression of Dnmt 1 in tissues from adult porcine animals.

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Evaluation of HalcyonTM Fast kV CBCT effectiveness in radiation therapy in cervical cancer patients of childbearing age who performed ovarian transposition (난소전위술을 시행한 가임기 여성의 자궁경부암 방사선치료 시 난소선량 감소를 위한 HalcyonTM Fast kV CBCT의 유용성 평가 : Phantom study)

  • Lee Sung Jae;Shin Chung Hun;Choi So Young;Lee Dong Hyeong;Yoo Soon Mi;Song Heung Gwon;Yoon In Ha
    • The Journal of Korean Society for Radiation Therapy
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    • v.34
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    • pp.73-82
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    • 2022
  • Purpose: The purpose of this study is to evaluate the effectiveness of reducing the absorbed dose to the ovaries and the quality of the CBCT image when using the HalcyonTM Fast kV CBCT of cervical cancer patients of child-bearing age who performed ovarian transposition Materials and Methods : Contouring of the cervix and ovaries required for measurement was performed on the computed tomography images of the human phantom (Alderson Rando Phantom, USA), and three Optically Stimulated Luminescence Dosimeter(OSLD) were attached to the selected organ cross-section, respectively. In order to measure the absorbed dose to the cervix and ovaries in the TruebeamTM pelvis mode (Hereinafter referred to as TP), The HalcyonTM Pelvis mode (Hereinafter referred to as HP) and The HalcyonTM Pelvis Fast mode (Hereinafter referred to as HPF), An image was taken with a scan range of 17.5 cm and also taken an image that reduced the Scan range to 12.5cm. A total of 10 cumulative doses were summed, It was replaced with a value of 23 Fx, the number of cervical cancer treatments, and compared In additon, uniformity, low contrast visibility, spatial resolution, and geometric distortion were compared and analyzed using Catphan 504 phantom to compare CBCT image quality between equipment. Each factor was repeatedly measured three times, and the average value was obtained by analysing with the Doselab (Mobius Medical Systems, LP. Versions: 6.8) program. Results: As a result of measuring absorbed dose by CBCT with OSLD, TP and HP did not obtain significant results under the same conditions. The mode showing the greatest reduction value was HPF versus TP. In HPF, the absorbed dose was reduced by 39.8% in the cervix and 19.8% in the ovary compared to the TP in the scan range of 17.5 cm. the scan range was reduced to 12.5 cm, absorbed dose was reduced by 34.2% in the cervix and 50.5% in the ovary. In addition, result of evaluating the quality of the image used in the above experiment, it complied with the equipment manufacturer's standards with Geometric Distortion within 1mm (SBRT standard), Uniformity HU, LCV within 2.0%, Spatial Resolution more than 3 lp/mm. Conclusion: According to the results of this experiment, HalcyonTM can select more various conditions than TruebeamTM in treatment of fertility woman who have undergone ovarian Transposition , because it is important to reduce the radiation dose by CBCT during radiation therapy. So finally we recommend HalcyonTM Fast kV CBCT which maintains image quality even at low mAs. However, it is consider that the additional exposure to low doses can be reduced by controlling the imaging range for patients who have undergone ovarian transposition in other treatment machines.

Effect of Di(2-ethyl hexyl)phthalate(DEHP) on the Onset of Puberty in Female Rat (암컷 흰쥐의 사춘기 개시에 미치는 di(2-ethyl hexyl)phthalate(DEHP)의 효과)

  • Lee, Kyeung-Yeup;Lee, Sung-Ho
    • Development and Reproduction
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    • v.10 no.2
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    • pp.147-154
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    • 2006
  • Phthalates such as di(2-ethyl hexyl)phthalate(DEHP) are industrial chemicals with wide-ranging human exposures because of their use in plastics and other common consumer products. Consequently, their adverse effects as endocrine disruptor in the reproductive physiology of both laboratory rodents and human have been studied extensively. The present study was undertaken to examine whether prepubertal exposure to DEHP affects on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. DEHP(100mg/kg/day) was administered daily from postnatal day 25(PND 25) through the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the next day. Gross anatomy and weight of reproductive tissues were compared to test the DEHP's effects on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in the tissues. Specific radioimmunoassay was carried out to measure serum LH levels. To determine the transcriptional changes in progesterone receptor(PR), total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a result, delayed VO was shown in the DEHP group(PND $37.3{\pm}0.7$) compared to the control group(PND $35.3{\pm}0.7$; p<0.05). DEHP treatment significantly decreased the wet weight of ovaries and uteri compared to the control group(p<0.05). Interestingly, elevation of serum LH levels was shown in the DEHP group(p<0.05). Graafian follicles and corpora lutea were observed only in the ovaries from the control animals. Numerous primary, secondary follicles and small atretic follicles were observed in the ovaries from DEHP-treated animals. Similarly, hypotrophy of luminal and glandular uterine epithelium was found in the DEHP-treated group. These effects were probably due to the inhibitory effects of DEHP on the synthesis and secretion of estrogen from granulosa cells. In the semiquantitative RT-PCR studies, the transcriptional activities of PR in both ovary(p<0.05) and uterus(p<0.01) from DEHP-treated animals were significantly lower than those from the control animals. The present studies demonstrated that the acute exposure to DEHP during the critical period of prepubertal stage could inactivate the reproductive system resulting delayed puberty in female rats.

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Effects of Anabolic Steroids of Pork on Proliferation and Differentiation of Myogenic Satellite Cell (돼지 고기의 아나볼릭 스테로이드가 Myogenic Satellite Cell의 증식과 분화에 미치는 영향)

  • Lee, Dong-Mok;Lee, Ki-Ho;Cheon, Yong-Pil;Chun, Tae-Hoon;Choi, In-Ho
    • Food Science of Animal Resources
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    • v.30 no.5
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    • pp.842-850
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    • 2010
  • Sex steroids are known to be involved in skeletal muscle development (anabolic effect) and are frequently used in medicines. It has been known that pork contains a variety of steroids that are mainly synthesized in the gonads (testis and ovary). Thus, the present study was conducted to evaluate the effects of anabolic steroids of pork on the proliferation and differentiation of myogenic satellite cells (MSC). Three different methods (M1, M2, and M3) were developed for the isolation and purification of steroids from porcine tissues. Among three extraction methods that we developed, M3 was the best method with respect to the quantities of steroids and the induction of MSC proliferation. Hormonal analysis showed that the steroid hormone levels were the highest in muscle and fat of intact male than those of castrated males and females. In addition, the highest serum levels of nandrolone and testosterone were detected in intact males, whereas estrone and $17{\beta}$-estradiol levels were similar in the entire experimental serum samples. Expression of androgen receptor (AR), myoD, desmin, and myogenin in bovine muscle cells were significantly up-regulated by the treatment of steroid extracts. The highest increas of myogenin and AR mRNA abundance were observed in the MSCs treated with M3 extract (p<0.001). Altogether, the present research showed the positive effect of steroids on MSC proliferation and differentiation in vitro. These results would certainly imply a beneficial effect of pork consumption on human muscle development.

Role of Ghrelin in the Control of Reproductive Endocrine Function (포유류 생식 내분비 기능 조절에서 Ghrelin의 역할)

  • Lee, Sung-Ho
    • Development and Reproduction
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    • v.13 no.4
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    • pp.207-215
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    • 2009
  • Numerous factors can affect the activities of hypothalamus-pituitary-gonad (HPG) hormonal axis, resulting in alteration of reproductive capacity or status such as onset of puberty and menopause. Soon after the finding of leptin, a multifunctional hormone secreted from adipocytes, a close relationship between reproduction and body energy balance have been manifested. Ghrelin, another multifunctional hormone from gastrointestinal tract, is an endogenous ligand of growth hormone secretagogue receptor (GHSR), and is thought to be a counterpart of leptin in the regulation of energy homeostasis. As expected, ghrelin can also modulate the reproductive capacity through the modulation of activities of HPG axis. This paper summarizes the current knowledge on the discovery, gene structures, tissue distribution and roles of ghrelin and GHSRs in mammalian reproduction in particular modulation of reproductive hormone secretion in HPG axis. Like POMC gene expression in pituitary gland, preproghrelin gene can generate a complex repertoire of transcripts which further undergo alternative splicing and posttranslational modifications. Concerning the roles of preproghrelin gene products in the control of body physiology except energy homeostasis, limited knowledge is available so far. Several lines of evidence, however, show the interplay of ghrelin between metabolism and reproduction. In rat and human, the distribution of ghrelin receptor GHSRs (GHSR1a and GHSR1b) has been confirmed not only in the hypothalamus and pituitary which were originally postulated as target of ghrelin but also in the testis and ovary. Expression of the preproghrelin gene in the brain and gonads was also verified, suggesting the local role (s) of ghrelin in HPG axis. Ghrelin might play a negative modulator in the secretions of hypothalamic GnRH, pituitary gonadotropins and gonadal steroids though the action on pituitary is still questionable. Recent studies suggest the involvement of ghrelin in regulation of puberty onset and possibly of menopause entry. It is now evident that ghrelin is a crucial hormomal component in 'brain-gut' axis, and is a strong candidate links between metabolism and reproduction. Opposite to that for leptin, ghrelin signaling is likely representing the 'hunger' state of body energy balance and is necessary to avoid the energy investment into reproduction which has not a top priority in maintaining homeostasis. Further researches are needed to gain a deep insight into the more precise action mechanism and role of ghrelin in reproduction, and to guarantee the successful biomedical applications.

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