• Title/Summary/Keyword: Differential staining

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Differential Expression of miR-34c and Its Predicted Target Genes in Testicular Tissue at Different Development Stages of Swine

  • Zhang, Xiaojun;Zhao, Wei;Li, Chuanmin;Yu, Haibin;Qiao, YanYan;Li, Aonan;Lu, Chunyan;Zhao, Zhihui;Sun, Boxing
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1532-1536
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    • 2015
  • To verified the target genes of miR-34c, bioinformatics software was used to predict the targets of miR-34c. Three possible target genes of miR-34c related to spermatogenesis and male reproductive development: zinc finger protein 148 (ZNF148), kruppel-like factor 4 (KLF4), and platelet-derived growth factor receptor alpha (PDGFRA) were predicted. Then, the expression of miR-34c and its target genes were detected in swine testicular tissue at different developmental stages by quantitative polymerase chain reaction. The results suggested that the expression of PDGFRA has the highest negative correlation with miR-34c. Then immunohistochemical staining was done to observe the morphology of swine testicular tissue at 2-days and 3, 4, 5-months of age, which indicated that PDGFRA was mainly expressed in the support cells near the basement membrane during the early development stages of testicular tissue, but that the expression of PDGFRA was gradually reduced in later stages. Therefore, western blot analyzed that the highest expression of PDGFRA was generated in 2-days old testicular tissues and the expression levels reduced at 3 and 4-months old, which correlated with the results of immunohistochemical staining. In conclusion, PDGFRA is a target gene of miR-34c.

Aberrant Distributions of ICM Cells in Bovine Blastocysts Produced by Somatic Cell Nuclear Transfer

  • D. B. Koo;Y. K. Kang;Park, Y. H.;Park, J. S.;Kim, H. N.;D. S. Son;Y. M. Han;Lee, K. K.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.20-20
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    • 2001
  • It has been reported that cloning cattle is inefficient. One of the problems was placental abnormality, finally resulting in fetal mortality after transfer of nuclear transfer (NT) bovine embryos. This study was focused on the allocations of embryonic cells to the inner cell mass (ICM) or to the trophectoderm(TE) in NT bovine blastocysts. Somatic cells were derived from a Day 45 fetus of gestation, individually transferred into enucleated oocytes and developed to the blastocyst stage in vitro. Differential staining was used to assess the qualify of blastocysts derived from NT, IVF and in vivo. Development rate of NT embryos to blastocysts (25.0%, 41/164) was similar to that of IVF embryos (28.7%, 49/171). The total cell number of NT blastocysts (101.3$\pm$45.9) was not different compared with that of IVF embryos (107.9$\pm$34.2, P>0.05), but was lower than in vivo embryos (122.5$\pm$21.6, P<0.05). Ratio of ICM/total cells was higher in NT embryos (51.6$\pm$ 18.6%) than in IVF and in vivo embryos (42.3$\pm$ 15.3% and 34.9$\pm$8.9%, respectively) (P<0.05). Most IVF (56.8%, 25/44) and in vivo blastocysts(80.8%, 21/26) was distributed in the proportion of ICM/total cells ranging from 20 to 40% group. However, most NT blastocysts was biased in the 40-60%(34.1%, 15/44) and >60% (31.8%, 14/44) groups. Our findings suggest that placental abnormalities or early fetal losses in the present cloning system may be due to aberrant allocation of NT embryos to the ICM cells.

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Differential Effects of Nongenotoxic and Genotoxic Carcinogen on Cell Proliferation and c-Jun Expression in the Rat Liver Initiated with Diethylnitrosamine

  • Kim, Hye-Jin;Kim, Jong-Won;Hong, Jin-Tae;Nam, Ki-Taek;Kim, Dae-Joong
    • Environmental Mutagens and Carcinogens
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    • v.19 no.2
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    • pp.89-94
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    • 1999
  • Cell proliferation and c-Jun expression pattern in liver exposed by nongenotoxic carcinogens phenobarbital (PB) and clofibrate, and genotoxic carcinogen 2-amino-3-methylimidazo [4,5-f] quinoline (IQ) were investigated to see whether differential effects of genotoxic and non-genotoxic carcinogens on the development of neoplastic foci may be related to differential effect on cell proliferation. Male F344 rats were initially given a single intraperitioneal injection of diethylnitrosamine (200 mg/kg body weight), and 2 weeks later, animals were fed diets containing 0.03% IQ or 0.5% CE or 0.05% PB or basal diet as a control for 6 weeks. All rats were subjected to the two-thirds partial hepatectomy (PH) at week 3. Sequential sacrifice of rats was performed until 8 weeks. Cell proliferation was examined by immunohistochemical staining of bromodeoxyuridine and c-Jun expression was determined by northern blotting. The increase of cell proliferation rate after PH was significant in the rats fed 0.05% IQ and continued until 8 weeks, while the increase was not significant in the rats fed phenobarbital and clofibrate compared to that in the rats fed control diet. mRNA level of c-Jun in the liver treated with IQ was about 7 fold higher than that of control and peak at 5 hours after rH. In the liver treated with CE, mRNA level of c-Jun was 3-4 fold higher than that of control and the highest level of mRNA of c-Jun was seen at 24 hours after PH. These results show that differential effects of genotoxic and non-genotoxic carcinogens on the development of neoplastic foci may be related to differential effect on cell proliferation pattern.

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The Comparison of Methods Processing Cells Recovered by Bronchoalveloar Lavage (기관지폐포세척액에서 세포수 측정을 위한 처리방법의 비교)

  • Park, Jae-Seuk;Kim, Jae-Yeal;Lee, Gwi-Lae;Yoo, Chul-Gyu;Han, Sung-Koo;Shim, Young-Soo;Kim, Young-Whan
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.1
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    • pp.176-183
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    • 1998
  • Background: The total and differential cell count of bronchoalveolar lavage(BAL) fluid are useful assessing activity, prognosis and response to therapy in diffuse interstitial lung disease. But controversy exist as to the appropriate method in processing BAL fluid. Therefore we investigated the effect of gauze filtration, centrifugation and different storage time of BAL fluid on the total and differential cell count. Methods: We obtained BAL fluid from 6 persons with no active lung lesion and divided pooled BAL fluid into several siliconized glass tubes and filtered through 0, 1, 2, 4 folds of cotton guaze(pore size: 1mm), and compared total cell count using hemocytometer after trypan blue staining and differential cell count after Wright-Giemsa staining of cytocentrifuged preparations. And we also counted total and differential cell count after centrifugation(400g for 30 min) and various storage time(2hr, 24hr, and 48hr). Results: There was no difference in total and differential cell count according to folds of gauze filtraion. But without gauze filtration, mucus threads that hampered total and differential cell count were found in 2 cases (33%). Centrifugation resulted in loss of total cell count($24{\pm}18%$) without change in differential cell count. There was no change in total cell count after 2hr storage but significant cell loss was found after 24hr storage time(24hr : $28{\pm}21%$, 48hr : $41{\pm}24%$). However there was no change in differential cell count with 48hr storage time. Conclusion: Total and differential cell count of BAL fluid may be best performed after cotton gauze filtration without centrifugation and within 2 hours.

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A Trial of Screening of Genes Involved in Odontoblasts Differentiation from Human Dental Pulp Stem Cells

  • Park, Yoon-Kyu;Kim, Hyun-Jin
    • International Journal of Oral Biology
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    • v.37 no.4
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    • pp.167-173
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    • 2012
  • This study investigated the genes involved in the differentiation of odontoblasts derived from human dental pulp stem cells (hDPSCs). hDPSCs isolated from human tooth pulp were validated by fluorescence activated cell sorting (FACS). After odontogenic induction, hDPSCs were analyzed investigated by Alizaline red-S staining, ALP assay, ALP staining and RT-PCR. Differential display-polymerase chain reaction (DD-PCR) was performed to screen differentially expressed genes involved in the differentiation of hDPSCs. By FACS analysis, the stem cell markers CD24 and CD44 were found to be highly expressed in hDPSCs. When hDPSCs were treated with agents such as ${\beta}$-glycerophosphate (${\beta}$-GP) and ascorbic acid (AA), nodule formation was exhibited within six weeks. The ALP activity of hDPSCs was found to elevate over time, with a detectable up-regulation at 14 days after odontogenic induction. RT-PCR analysis revealed that dentin sialophosphoprotein (DSPP) and osteocalcin (OC) expression had increased in a time-dependent manner in the induction culture. Through the use of DD-PCR, several genes were differentially detected following the odontogenic induction. These results suggest that these genes may possibly be linked to a variety of cellular process during odontogenesis. Furthermore, the characterization of these regulated genes during odontogenic induction will likely provide valuable new insights into the functions of odontoblasts.

Is Immunohistochemical Sex Hormone Binding Globulin Expression Important in the Differential Diagnosis of Adenocarcinomas?

  • Bulut, Gulay;Kosem, Mustafa;Bulut, Mehmet Deniz;Erten, Remzi;Bayram, Irfan
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8203-8210
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    • 2016
  • Adenocarcinomas (AC) are the most frequently encountered carcinomas. It may be quite challenging to detect the primary origin when those carcinomas metastasize and the first finding is a metastatic tumor. This study evaluated the role of sex hormone binding globulin (SHBG) positivity in tumor cells in the subclassification and detection of the original organ of adenocarcinomas. Between 1994 and 2008, 64 sections of normal tissue belonging to ten organs, and 116 cases diagnosed as adenoid cystic carcinoma and mucoepidermoid carcinoma of the salivary gland, lung adenocarcinoma, invasive ductal carcinoma of the breast, adenocarcinoma of stomach, colon, gallbladder, pancreas and prostate, endometrial adenocarcinoma and serous adenocarcinoma and mucinous adenocarcinoma of the ovary, were sent to the laboratory at the Department of Pathology at the Yuzuncu Yil University School of Medicine, where they were stained immunohistochemically, using antibodies against SHBG. The SHBG immunoreactivity in both the tumor cells and normal cells, together with the type, diffuseness and intensity of the staining were then evaluated. In the differential diagnosis of the adenocarcinomas of the organs, including the glandular structures, impressively valuable results are encountered in the tumor cells, whether the SHBG immunopositivity is evaluated alone or together with other IHC markers. Further extensive research with a larger number of cases, including instances of cholangiocarcinoma and cervix uteri AC [which we could not include in the study for technical reasons] should be performed, in order to appropriately evaluate the role of SHBG in the differential diagnosis of AC.

Meningeal Solitary Fibrous Tumor

  • Lee, Jong-Myong
    • Journal of Korean Neurosurgical Society
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    • v.37 no.3
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    • pp.232-234
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    • 2005
  • We report a rare case of a patient with meningeal solitary fibrous tumor. A 60-year-old woman presented with right leg monoparesis. Brain magnetic resonance imaging demonstrates a well enhancing huge mass, located in left parietal lobe. Cerebral angiography demonstrating increased vascularity in area of the tumor, which had feeder vessels extending from the internal carotid artery and external carotid artery. A presumptive diagnosis of meningioma or hemangiopericytoma was considered. At surgery, the consistency was firm and had destroyed the dura and skull. A gross total resection was performed. Immunohistochemically, tumor was strongly, and widely, positive for CD34 and vimentin. There was no staining for epithelial membrane antigen(EMA), S-100 protein, cytokeratin, and glial fibrillary acidic protein (GFAP). Differential diagnosis of intracranial solitary fibrous tumor includes fibroblastic meningioma, meningeal hemangiopericytoma, neurofibroma, and schwannoma.

Olfactory Groove Schwannoma

  • Prak, Ji-Hwan;Kim, Tae-Young;Park, Jong-Tae;Kim, Jong-Moon
    • Journal of Korean Neurosurgical Society
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    • v.39 no.2
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    • pp.156-158
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    • 2006
  • We present a case of olfactory schwannoma in a 16-year-old boy with headache and diplopia. Brain computed tomography[CT] scan and magnetic resonance[MR] imaging showed a huge mass in the subfrontal area resembling an olfactory groove meningioma. We performed a bifrontal craniotomy and found out the mass was attached to cribriform plate but was not related to the olfactory tract or bulb. The histopathological diagnosis of schwannoma was confirmed by immunohistochemical staining for S-100, vimentin and others. We describe the clinical manifestations, radiological characteristics, histological aspects, and differential diagnosis of this tumor with literature review.

Partial Mitotic Synchronization and Giemsa G-banding in Allium wakegi

  • Bong Bo Seo
    • Journal of Plant Biology
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    • v.38 no.1
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    • pp.33-38
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    • 1995
  • Hydroxyurea (HU), a DNA synthesis inhibitor, was tested as synchronizing agent in root-tip meristem of Allium wakegi. Roots were treated with 2.5mM HU for 14 h to accumulate meristem root-tip cells at the G1/S interface. After release from HU block, the cells re-entered the cell cycle with a high degree of synchrony. Synchronized mitotic frequency of A. wakegi was 22.7%, which was about 3.9 times as high as that of the control. The highest metaphase index(23.0%) was obtained when, 6 h after release from the HU block, the roots were treated with 0.05% colchicine for 2 h. Modifying various Giemsa staining protocols defined for animals and a few plant species, G-bands were visualized at prometaphase and metaphase chromosomes of A. wakegi. The higher degree of chromosome condensation, the less differential bands could be resolved. This is the first demonstration introduced partial mitotic synchronization into G-banding in plant.

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Differential Diagnosis of Pleural Mesothelioma and Metastatic Adenocarcinoma by Immunohistochemistry (면역조직화학염색법을 이용한 흉막의 악성중피종과 전이성 선암의 감별진단)

  • Ko, Kyung-Haeng;Park, Chang-Min;Rim, Myung-Soo;Kim, Yoo-Il;Jang, Il-Gweon;Hwang, Joon-Hwa;Lim, Sung-Chul;Kim, Young-Chul;Park, Kyung-Ok;Park, Chang-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.4
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    • pp.478-487
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    • 1999
  • Background : Differential diagnosis of pleural malignant mesothelioma from secondary metastatic adenocarcinoma is often difficult. A variety of pathologic techniques have been developed to make a differential diagnosis of carcinoma from mesothelioma. Immunohistochemistry detecting diverse antigenic substances such as CEA, Leu-M1, Bn-3, S-100 protein, vimentin, CK and EMA has been claimed to be of value as a panel in the differential diagnosis of adenocarcinoma from mesothelioma. The aim of this study was to investigate the suitable antibodies to distinguish mesothelioma from metastatic adenocarcinoma and establish candidate markers in a panel. Methods : Complete, one-hour immunohistochemical staining using antibodies against cytokeratin (CK), epithelial membrane antigen(EMA), S-100 protein, vimentin, B72-3, Leu-M1, and carcino-embryonic antigen(CEA) was applied to cell blocks from 7 mesotheliomas and 7 adenocarcinomas which were confirmed by electron microscopic and histpathologic methods. Results : All adenocarcinomas and 71.4% of mesotheliomas expressed the cytokeratin and EMA. S-100 protein and vimentin were expressed in 57.1% and 42.9% of mesotheliomas and 14.3% and 28.5% of adenocarcinomas, respectively. B72-3 was expressed in all adenocarcinomas, but in none of mesotheliomas. Leu-M1 was positive in 71.4% of the adenocarcinoma and 14.3% of the mesotheliomas. CEA was positive in all adenocarcinomas and 42.9% of mesotheliomas. Leu-M1 and B72-3 were coexpressed in 71.4% of adenocarcinomas but in none of mesothelioma. B72-3 and CEA were coexpressed in all adenocarcinomas, but in none of mesotheliomas. Conclusion : We concluded that B72-3 immunohistochemistry or panel staining of B72-3 and CEA could be recommanded for the differential diagnosis of pleural mesothelioma from metastatic adenocarcinoma.

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