• 제목/요약/키워드: Chromatin structure

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A primo vessel-like structure in a dog with inflammatory pseudotumor

  • Cho, Sung-Jin;Hong, Sun-Hwa;Han, Sang-Jun;Kim, Ok-Jin
    • 한국동물위생학회지
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    • 제35권1호
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    • pp.77-82
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    • 2012
  • Inflammatory pseudotumor (IPT) is a term defining a mass characterized microscopically by a proliferation of bland mesenchymal spindle cells infiltrated by diffuse mixed inflammatory cells with a predominance of plasma cells and lymphocytes. Here, we show the primo vessel-like structure of the primo-vascular system (PVS) in a dog with IPT. A 6-years old male Mongrel dog was diagnosed with an abnormal mass (diameter 5.5 cm, weight 22 g) near left preputial area. The dog was submitted to the surgical detectomy of the mass. During the surgical operation, we observed primo vessel-like material. After fixations, the masses appeared macroscopically as lipoid-like, firm, white to grey masses, measuring $5{\times}8cm$. Histologically, cellular infiltration into the muscular layers was frequently seen. The mesenchymal proliferation remained the main component of the mass and was composed of myofibroblastic-like spindle cells characterized by globular, irregular nuclei containing open chromatin and a prominent nucleolus. On the basis of the histopathologic lesions, the subcutaneous mass was diagnosed as IPT. Also, we detected a primo vessel-like structures in some areas of the IPT tissues. These were observed as novel thread-like structures and bundle of tubular structures. To our knowledge, this report is the first case of primo vessel-like structure in a dog with IPT.

Magnetic Orientations of Bull Sperm Treated by DTT or Heparin

  • Suga, D.;Shinjo, A.;Kumianto, E.;Nakada, T.
    • Asian-Australasian Journal of Animal Sciences
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    • 제13권1호
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    • pp.10-18
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    • 2000
  • This paper describes the magnetic orientation of the intact and demembranated bull sperm treated by DTT or heparin in a 5,400 G static field. Semen samples collected from four bulls (Japanese Black) were mixed to the same sperm density. One percentage triton X-100 was used to extract the plasma membrane. The intact and demembranated sperm suspensions were treated with 20, 200, 2,000 mM DTT, 100, 1,000 or 10,000 units heparin solutions at $4{^{\circ}C}$ for 6 days. The decondensation of the sperm nuclei treated by DTT or heparin was examined by measuring the sperm head area at 1, 3, and 6 days. After measuring the area, each sperm sample was exposed to a 5,400 G static magnetic field generated by Nd-Fe-B permanent magnets for 24 hours at room temperature. Results showed that the decondensation of bull sperm nuclei was not induced by the heparin treatment, however, incomplete decondensation was induced by the DTT treatment. During the magnetic orientation, bull sperms treated by DTT or heparin had low percentages of long axis perpendicular to the magnetic lines of force. However, different aspects were obtained for long axis perpendicular orientations following treatment of DTT or heparin. Through the DTT treatment, the decline of long axis perpendicularly oriented percentages was due to the increase of long axis parallel orientation with the head of the flat plane perpendicular to the magnetic lines of force, whereas, using the heparin treatment, the decline of long axis perpendicular orientation was due to the increment of long axis parallel orientation with the head of the flat plane parallel to the magnetic lines of force. Also, percentages of the head of the flat plane perpendicular were decreased by the heparin treatment. These findings suggest that maintaining the structure of protamine in the chromatin is necessary for the sperm head to orient with its flat plane perpendicular, and maintaining the disulfide bond in the chromatin is necessary for the long axis of sperm to orient perpendicularly.

소 체내포 핵이식에 의한 핵-세포질 상호작용에 관한 연구 (Study on Nucleo-Cytoplasmic Interaction by Somatic Cell Nuclear Transfer in Bovine)

  • 정희태;최종엽;박춘근;김정익;민동미
    • 한국수정란이식학회지
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    • 제15권1호
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    • pp.23-31
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    • 2000
  • This study was conducted to investigate the effects of quiescent treatment of donor cells and activation treatment time of recipient cytoplasm on nuclear remodeling and in vitro development of somatic cell-cloned bovine embryos. Serum starved, confluent and nonquiescent cycling adult skin cells were teansferred into enucleated oocytes. Nuclear transfer oocytes were activated at 30 min, 1 and 2 hrs after electrofusion. Some nuclear transfer embryos(23% to 35%) extruded a polar body, which was not affected by quiescent treatment of donor cells and activiation time of recipient cytoplasm. About 68% of nuclear transfer embryos fused with a serum starved cells has a chromatin clump, but which was not different from embryos fused with confluent(51%) and nonquiescent(47%) cells. The proportion of embryos with a single chromatin clump was sightly increased when nuclear transfer embryos were activated within 30 min after fusion(69%) compared to those were activated at 1 and 2 hrs after fusion, but there was not significantly different. Development rates to the blastocyst stage were 8.6% and 15.9% when serum starved and confluent cells were transferred, which were higher than that of control group. Developmental rate to the blastocyst stage was higher in embryos were activated within 30 min after fusion (17.3%) compared to those of embryos were activated at 1 and 2 hrs after fusion (P<0.05). From the present result, it is suggested that quiescent treatment of donor cells and activation time of recipient cytoplasm can affect the in vitro development. Quiescent plasm activation within 30 min after fusion could increase the number of embryos with a normal chromation structure, which results in increased in vitro development.

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Evaluation of sperm DNA fragmentation using multiple methods: a comparison of their predictive power for male infertility

  • Javed, Aamir;Talkad, Muralidhar Srinivasaih;Ramaiah, Manjula Kannasandra
    • Clinical and Experimental Reproductive Medicine
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    • 제46권1호
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    • pp.14-21
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    • 2019
  • Objective: The usual seminal profile has been customarily used for diagnosing male infertility based on an examination of semen samples. However, sperm DNA fragmentation has also been causally linked to reproductive failure, suggesting that it should be evaluated as part of male infertility assessments. To compare the ability of the five most widely utilized methodologies of measuring DNA fragmentation to predict male infertility and reactive oxygen species by Oxisperm kit assay. Methods: In this case-control study, which received ethical committee approval, the participants were divided into fertile and infertile groups (50 patients in each group). Results: The alkaline comet test showed the best ability to predict male infertility, followed by the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay, the sperm chromatin dispersion (SCD) test, and the sperm chromatin structure assay (SCSA), while the neutral comet test had no predictive power. For our patient population, the projected cut-off point for the DNA fragmentation index was 22.08% using the TUNEL assay, 19.90% using SCSA, 24.74% using the SCD test, 48.47% using the alkaline comet test, and 36.37% using the neutral comet test. Significant correlations were found between the results of the SCD test and those obtained using SCSA and TUNEL (r = 0.70 and r = 0.68, respectively; p< 0.001), and a statistically significant correlation was also found between the results of SCSA and the TUNEL assay (r = 0.77, p< 0.001). Likewise, the results of the alkaline comet test showed significant correlations with those of the SCD, SCSA, and TUNEL tests (r = 0.59, r = 0.57, and r = 0.72, respectively; p< 0.001). Conclusion: The TUNEL assay, SCSA, SCD, and the alkaline comet test were effective for distinguishing between fertile and infertile patients, and the alkaline comet test was the best predictor of male infertility.

Downregulation of JMJD2a and LSD1 is involved in CK2 inhibition-mediated cellular senescence through the p53-SUV39h1 pathway

  • Park, Jeong-Woo;Bae, Young-Seuk
    • BMB Reports
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    • 제55권2호
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    • pp.92-97
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    • 2022
  • Lysine methylation is one of the most important histone modifications that modulate chromatin structure. In the present study, the roles of the histone lysine demethylases JMJD2a and LSD1 in CK2 downregulation-mediated senescence were investigated. The ectopic expression of JMJD2a and LSD1 suppressed the induction of senescence-associated β-galactosidase activity and heterochromatin foci formation as well as the reduction of colony-forming and cell migration ability mediated by CK2 knockdown. CK2 downregulation inhibited JMJD2a and LSD1 expression by activating the mammalian target of rapamycin (mTOR)-ribosomal p70 S6 kinase (p70S6K) pathway. In addition, the down-regulation of JMJD2a and LSD1 was involved in activating the p53-p21Cip1/WAF1-SUV39h1-trimethylation of the histone H3 Lys9 (H3K9me3) pathway in CK2-downregulated cells. Further, CK2 downregulation-mediated JMJD2a and LSD1 reduction was found to stimulate the dimethylation of Lys370 on p53 (p53K370me2) and nuclear import of SUV39h1. Therefore, this study indicated that CK2 downregulation reduces JMJD2a and LSD1 expression by activating mTOR, resulting in H3K9me3 induction by increasing the p53K370me2-dependent nuclear import of SUV39h1. These results suggest that CK2 is a potential therapeutic target for age-related diseases.

세포의 휴면처리가 소 태아섬유아세포 유래 핵이식란의 핵상변화와 체외발육에 미치는 영향 (Effect of Quiescent Treatment on Nuclear Remodeling and In Vitro Development of Nuclear Transfer Embryos Derived from Bovine Fetal Fibroblast Cells)

  • 최종엽;권대진;김정익;박춘근;양부근;정희태
    • 한국가축번식학회지
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    • 제24권2호
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    • pp.217-222
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    • 2000
  • 본 연구는 세포의 휴면처리가 소 태아섬유아세포 유래 핵이식란의 핵형변화와 체외발육에 미치는 영향을 검토하였다. 임신 3∼4 개월령 한우 웅성 태아의 피부세포를 채취하여 계대배양 후 동결하였다가 핵이식 전에 혈청기아처리 또는 confluency방법으로 휴면처리를 하여 미수정란의 탈핵세포질에 이식하였다. 전기융합과 활성화처리 후 7∼9 일간 체외배양하여 발육능을 검토하였으며, 일부는 whole-mount 법으로 고정하여 염색질 구조를 관찰하였다. 복제란의 극체방출율은 혈청기아처리구와 confluence 구에서 각각 24.5% 와 20.3% 로 무처리구(36.0%) 에 비해 낮은 경향을 보였다. 활성화 후 1개의 염색질괴를 갖는 복제란은 혈청기아처리구(50.9%)와 confluence구 (49.2%)가 무처리구 (40.0%) 보다 높은 경향을 보였다. 극체방출에 따른 염색질의 구조는 극체 미방출구에서 정상적인 1개의 염색질괴를 갖는 핵이식란이 60.5% 로, 극체 방출구 (4.7%) 에 비하여 유의적으로 높았다 (P<0.01). 배반포기 발육율은 혈청기아처리구 (21.7%) 와 confluence (20.9%) 가 무처리구 (14.1%) 에 비하여 비교적 높게 나타났다. 본 연구의 결과 혈청기아처리나 confluency 방법에 의한 donor 세포의 휴면처리는 태아섬유아세포 유래 핵이식란의 비정상적인 핵형변화를 감소시켜 체외발육능을 향상시키는 것으로 사료된다.

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Histone H4-Specific Deacetylation at Active Coding Regions by Hda1C

  • Lee, Min Kyung;Kim, TaeSoo
    • Molecules and Cells
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    • 제43권10호
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    • pp.841-847
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    • 2020
  • Histone acetylation and deacetylation play central roles in the regulation of chromatin structure and transcription by RNA polymerase II (RNA Pol II). Although Hda1 histone deacetylase complex (Hda1C) is known to selectively deacetylate histone H3 and H2B to repress transcription, previous studies have suggested its potential roles in histone H4 deacetylation. Recently, we have shown that Hda1C has two distinct functions in histone deacetylation and transcription. Histone H4-specific deacetylation at highly transcribed genes negatively regulates RNA Pol II elongation and H3 deacetylation at inactive genes fine-tunes the kinetics of gene induction upon environmental changes. Here, we review the recent understandings of transcriptional regulation via histone deacetylation by Hda1C. In addition, we discuss the potential mechanisms for histone substrate switching by Hda1C, depending on transcriptional frequency and activity.

Effect of Activation Time on the Nuclear Remodeling and In Vitro Development of Nuclear Transfer Embryos Derived from Bovine Somatic Cells

  • Choi, J. Y.;J. W. Cho;D. J. Kwon;Park, C. K.;B. K. Yang;Kim, C. I.;H. T. Cheong
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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    • pp.6-6
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    • 2002
  • This study was conducted to investigate the effect of recipient activation time on the nuclear remodeling, chromatin structure, pronuclear formation and in vitro development of bovine nuclear transfer embryos derived from adult ear skin cells. Somatic cells were transferred to enucleated oocytes after quiescent treatments by serum starvation or culture to confluency. Nuclear transfer embryos were activated with a combination of Ca/sup 2+/-ionophore and cycloheximide at 1, 1.5, 2, 2.5, 3, and 5 h after electrofusion. (omitted)

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RNA Interference as a Plausible Anticancer Therapeutic Tool

  • Ramachandran, Puthucode Venkatakrishnan;Ignacimuthu, Savarimuthu
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권6호
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    • pp.2445-2452
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    • 2012
  • RNA interference has created a breakthrough in gene silencing technology and there is now much debate on the successful usage of RNAi based methods in treating a number of debilitating diseases. Cancer is often regarded as a result of mutations in genomic DNA resulting in faulty gene expression. The occurrence of cancer can also be influenced by epigenetic irregularities in the chromatin structure which leads to alterations and mutations in DNA resulting in cancer cell formation. A number of therapeutic approaches have been put forth to treat cancer. Anti cancer therapy often involves chemotherapy targeting all the cells in common, whereby both cancer cells as well as normal cells get affected. Hence RNAi technology has potential to be a better therapeutic agent as it is possible to deactivate molecular targets like specific mutant genes. This review highlights the successful use of RNAi inducers against different types of cancer, thereby paving the way for specific therapeutic medicines.

NMR characterization of SRG3 SWIRM Domain Mutant Proteins.

  • Koh, Woo-Hyoung;Kim, Min-Tae;Moon, Sun-Jin;Lee, Weon-Tae
    • 한국자기공명학회논문지
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    • 제13권1호
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    • pp.56-63
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    • 2009
  • SWIRM domain, a core domain of SRG3 is well conserved in SW13, RSC8, and MOIRA family proteins. To understand structural basis for cellular functions of the SWIRM domain, we have initiated biochemical and structural studies on SWIRM domain and mutants using gelfiltration chromatography, circular dichroism and NMR spectroscopy. The structural properties of the mutant SWIRM domains (K34A and M75A) have been characterized, showing that the structures of both wild-type and mutant proteins are a-helical conformation. The data conclude that mutations at interaction sites of its binding partner protein do not affect its secondary and tertiary structure.