• Title/Summary/Keyword: Tissue specificity

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Effects of Angiotensin Converting Enzyme Inhibition on Gene Expression of the Renin-Angiotensin System in Rats

  • Lee, Young-Rae;Lee, Mi-Young;Kim, Woon-Jung;Lee, Won-Jung
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.6
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    • pp.771-778
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    • 1998
  • To investigate interaction of angiotensin converting enzyme (ACE) inhibitor with local tissue renin- angiotensin system (RAS), changes in gene expression of the RAS components in various tissues in response to chronic administration of an ACE inhibitor, enalapril, were examined in Sprague-Dawley male rats. Enalapril was administered in their drinking water $(3{\sim}4\;mg/day)$ over 8 wk. Plasma and renal ACE activity increased significantly after 4 and 8 wk of enalapril treatment. Renin levels of the plasma and kidney of the enalapril-treated rats markedly increased after 4 wk and decreased thereafter, but still remained significantly higher than those of control rats. Kidney mRNA levels of renin markedly increased after 4 and 8 wk of enalapril treatment, but those of angiotensinogen and ANG II-receptor subtypes, $AT_{1A}$ and $AT_{1B}$, did not change significantly. The liver expressed genes for renin, angiotensinogen and $AT_{1A}$ receptor subtype, but $AT_{1B}$ receptor subtype mRNA was not detectable by RT-PCR. None of mRNA for these RAS components in the liver changed significantly by enalapril treatment. The hypothalamus showed mRNA expressions of renin, angiotensinogen, $AT_{1A}$ and $AT_{1B}$ receptor subtypes. $AT_{1A}$ receptor subtype mRNA was more abundant than $AT_{1B}$ receptor subtype in the hypothalamus as shown in the kidney. However, gene expression of the RAS components remained unchanged during 8-wk treatment of enalapril. In the present study, chronic ACE inhibition increased plasma and renal levels of ACE and renin, but did not affect mRNA levels of other RAS components such as angiotensinogen, ANG II receptor subtypes in the kidney. Gene levels of the RAS components in the liver and hypothalamus were not altered by chronic treatment of enalapril. These results suggest the differential expression of the RAS components in response to enalapril, and localized action and some degree of tissue specificity of enalapril.

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White Sport Syndrome Virus Disease of Shirmp and Diagnostic Methods

  • Zhan, Wen-Bin
    • Journal of Aquaculture
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    • v.15 no.1
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    • pp.7-13
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    • 2002
  • Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.

The Role of Thallium-201 Scintigraphy in Bone and Soft Tissue Tumor (근골격계 종양에서 탈륨 스캔의 역할)

  • Shin, Duk-Seop
    • Journal of Yeungnam Medical Science
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    • v.20 no.2
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    • pp.117-128
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    • 2003
  • Thallium-201 scintigraphy is used to discriminate the malignant bone tumor from the benign by qualitatively and quantitatively, and to predict the response of preoperative chemotherapy in osteosarcoma, by comparing the changes of thallium uptake ratio after chemotherapy to the tumor necrosis ratio. Thallium-201 scintigraphy scan should be done prior to surgical biopsy. PICKER Prism 2000 gamma camera with high resolution parallel hole collimator is usually used for scanning. The patient is injected with 2-3mCi of Tl-201 and the early phase is checked in 30 minutes and delayed phase in 3 hours. The scan images are visually evaluated by a blinded nuclear medicine physician. We could evaluate true positive, true negative, false positive and false negative by the comparison of results with those of biopsy, and calculate positive and negative predictive value(%), sensitivity(%), specificity(%) and diagnostic accuracy(%). For the quantitative analysis of thallium uptake, we drew the region of interest on the tumor side and contralateral normal side as mirror image, and calculated the uptake ratio with dividing the amount of gamma count in tumor side by normal side. We could calculate the percent changes of thallium uptake ratio in early and delayed phase, and compare them to the ratio of tumor necrosis. Thallium-201 scintigraphy proved as useful imaging study to discriminate malignant bone tumor from benign, but had exception in giant cell tumor and low grade malignant bone tumors. We can use T1-201 scan to differentiate the benign from the malignant tumor, and to evaluate the response of preoperative chemotherapy or radiotherapy, and to determine the residual tumor or local recurrence. For the better result, we need to have a more detail information about false positive cases and a more objective and quantitative reading technique.

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Identification of Viral RNA by Light Microscopic in situ Hybridization (광학현미경 In Situ Hybridization에 의한 Viral RNA 증명)

  • 최원기;주경웅;김석홍
    • Biomedical Science Letters
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    • v.2 no.2
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    • pp.249-255
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    • 1996
  • In this paper, a in situ hybridization(ISH) has been used to investigate the yield of viral RNA expression from each organ tissues. It is studied to establish a rapidly, specific diagnostic method detecting rabbit haemorrhagic disease virus(RHDV) RNA in 10% formalin-fixed, paraffin-em-bedded tissues of naturally RHDV-infected rabbits using oligonucleotide probe to be made by RHDV total sequences. Biotin was used as the oligonucleotide probe marker. in situ hybridization is detected the virus genome in the cells and tissue as specifically compared with others nucleic acid hybridization method. All ISH procedure of RHDV were completed to Mi-croProbe$^{TM}$ capillary action system within 1-2 hours. In this report, RHDV was distributed widely in the cytoplasm of liver cell and the cortex of kidney but lung tissue and medulla of kidney were showed to positive reaction at locally. Although not entirely free of technical limitations, nucleic acid identification holds advantages over other diagnostic tests, including exquisite sensitivity, specificity, interchangeability and speed. It is expected that, in the immediate future viral nucleic acid detection will be a prominent part of the methods used in histopathology.

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Serum Pleiotrophin Could Be an Early Indicator for Diagnosis and Prognosis of Non-Small Cell Lung Cancer

  • Du, Zi-Yan;Shi, Min-Hua;Ji, Cheng-Hong;Yu, Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.4
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    • pp.1421-1425
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    • 2015
  • Aims: Pleiotrophin (PTN), an angiogenic factor, is associated with various types of cancer, including lung cancer. Our aim was to investigate the possibility of using serum PTN as an early indicator regarding disease diagnosis, classification and prognosis, for patients with non-small cell lung cancer (NSCLC). Methods: Significant differences among PTN levels in patients with small cell lung cancer (SCLC, n=40), NSCLC (n=136), and control subjects with benign pulmonary lesions (n=21), as well as patients with different pathological subtypes of NSCLC were observed. Results: A serum level of PTN of 300.1 ng/ml, was determined as the cutoff value differentiating lung cancer patients and controls, with a sensitivity and specificity of 78.4% and 66.7%, respectively. Negative correlations between serum PTN level and pathological differentiation level, stage, and survival time were observed in our cohort of patients with NSCLC. In addition, specific elevation of PTN levels in pulmonary tissue in and around NSCLC lesions in comparison to normal pulmonary tissue obtained from the same subjects was also observed (n=2). Conclusion: This study suggests that the serum PTN level of patients with NSCLC could be an early indicator for diagnosis and prognosis. This conclusion should be further assessed in randomized clinical trials.

Genome-wide Analysis of Aberrant DNA Methylation for Identification of Potential Biomarkers in Colorectal Cancer Patients

  • Fang, Wei-Jia;Zheng, Yi;Wu, Li-Ming;Ke, Qing-Hong;Shen, Hong;Yuan, Ying;Zheng, Shu-Sen
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.1917-1921
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    • 2012
  • Background: Colorectal cancer is one of the leading causes of mortality worldwide. Genome wide analysis studies have identified sequence mutations causing loss-of-function that are associated with disease occurrence and severity. Epigenetic modifications, such DNA methylation, have also been implicated in many cancers but have yet to be examined in the East Asian population of colorectal cancer patients. Methods: Biopsies of tumors and matched non-cancerous tissue types were obtained and genomic DNA was isolated and subjected to the bisulphite conversion method for comparative DNA methylation analysis on the Illumina Infinium HumanMethylation27 BeadChip. Results: Totals of 258 and 74 genes were found to be hyper- and hypo-methylated as compared to the individual's matched control tissue. Interestingly, three genes that exhibited hypermethylation in their promoter regions, CMTM2, ECRG4, and SH3GL3, were shown to be significantly associated with colorectal cancer in previous studies. Using heatmap cluster analysis, eight hypermethylated and 10 hypomethylated genes were identified as significantly differentially methylated genes in the tumour tissues. Conclusions: Genome-wide methylation profiling facilitates rapid and simultaneous analysis of cancerous cells which may help to identify methylation markers with high sensitivity and specificity for diagnosis and prognosis. Our results show the promise of the microarray technology in identification of potential methylation biomarkers for colorectal cancers.

Establishment and characterization of porcine mammary gland epithelial cell line using three dimensional culture system (3차원 배양 시스템을 이용한 돼지 유선 상피 세포 주 특성과 설정)

  • Chung, Hak-Jae
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.10
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    • pp.551-558
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    • 2017
  • To study and validate tissue-specific promoters and vectors, it is important to develop cell culture systems that retain the tissue and species specificity. Such systems are attractive alternatives to transgenic animal models. This study established a line of porcine mammary gland epithelial cells (PMECs) from a primary culture based on the cellular morphology and mRNA levels of porcine beta-casein (CSN2). The selected PMECs were stained with the cytokeratin antibody, and were shown to express milk protein genes (CSN2, lactoferrin, and whey acidic protein). In addition, to confirm the acini structure of PMEC932-7 in 3D culture, live cells were stained with SYTO-13 dye, which binds to nucleic acid. The acini of these PMECs on matrigel were formed by the aggregation of peripheral cells and featured a hollow lumens. The system was demonstrated by testing the effects of the culture conditions to cell culture including cell density and matrigel methods of the PMECs. These results suggest that PMECs possess the genetic and structural features of mammary epithelial cells.

Estrogen Receptor Analysis in Fine Needle Aspirates and Frozen Sections from Human Breast Carcinomas (세침흡인 검사물을 이용한 유방암세포 에스트로젠수용체 분석 : 동결절편조직과의 비교)

  • Gong, Gyung-Yub;Ahn, Se-Hyun;Park, Kun-Choon;Choe, Ghee-Young;Yu, Eun-Sil;Lee, In-Chul
    • The Korean Journal of Cytopathology
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    • v.5 no.1
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    • pp.10-14
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    • 1994
  • The expression of sex steroid hormone receptors by neoplastic cells is an important predictor of response to hormone therapy. Thus, the selection of treatment modality is often based on the identification of receptors in tumor tissue. Various monoclonal antibodies of high specificity are now available for analyzing the estrogen receptor (ER). With these antibodies, biochemical enzyme immunoassay and immunohistochemistry using histologic sections have been used for ER analysis. We used fine needle aspirates from 15 human primary breast carinomas for the analysis of ERs. The semiquantitative receptor values obtained in cytologic specimens were correlated well with those from histologic specimens. The results of ER in fine needle aspirates were concordant with ER in histologic specimens(r=0.94). Only three cases showed a little difference in staining intensity and proportion of positive cells. Our results showed a good correlation between the receptor values determined in cytologic smears and those determined in tissue sections. It is suggested that measurement of the ER in cytologic smears may be a reliable technique which can be performed on aspiration cytologic samples.

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Development of Virulence Test Methods for Neck and Panicle Blast Disease (벼에 발생하는 목 도열병 및 이삭 도열병 검정을 위한 실험방법 개발)

  • Chi, Myoung-Hwan;Park, Sook-Young
    • Research in Plant Disease
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    • v.21 no.2
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    • pp.103-106
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    • 2015
  • Isolates of the rice blast fungus show a range of tissue-specificities infecting leaves, nodes, neck and panicles. Although neck and panicle blast cause significantly greater yield losses than the leaf blast, virulence tests of the blast isolates have been performed only rice leaves instead of neck and panicles. In this study, we have developed a virulence test method for neck and panicle blast. We selected three representative isolates from each of leaf, neck, and panicle blast. We observed that severe disease lesions developed on the neck and the panicles when the infected rice plants were incubated in a dew chamber for 48 h instead of 24 h when tested on leaves. Unlike the leaf blast, a typical lesion on the neck and panicles appeared after 14 days post-infection as opposed to 7 days with leaf blast. This method will be applied to examine tissue-specificity of the rice blast fungus isolates.

Peroxidase Activity during Leaf Infection of Mulberry (Morus alba L.) with Brown Leaf Spot Fungus Myrothecium roridum

  • Chattopadhyay Soumen;Krishnan Natraj;Maji Manas D.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.12 no.1
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    • pp.21-28
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    • 2006
  • Peroxidase activity was measured in brown leaf spot pathogen (Myrothecium roridum) inoculated potted mulberry (Morus alba) during pre-symptomatic to various symptom development stages and compared with corresponding healthy leaf tissues. The enzyme showed a pH optimum of 7.0 and the activity was linearly increased up to 15 min of incubation. The peroxidase had a broad substrate specificity and the rates of oxidation were in the rank of pyrogallol> guaiacol> ascorbate at pH 7.0. Catechol at 10 mM inhibited 89% of guaiacol-peroxidase and 76% pyrogallol-peroxidase activities, indicated higher non-specific peroxidation in pyrogallol dependent assay system in mulberry than guaiacol. The optimum requirement for the guaiacol dependent assay was 0.2 ml (${\approx}40-60{\mu}g$ equivalent of protein) of crude enzyme source. Excepting the 8th leaf from the apex, the peroxidase activity did not vary appreciably in different leaf positions. In pre-symptomatic phases, an initial (1 to 5 min) rise of peroxidase activity was noticed in inoculated leaves, and then maintained a plateau up to 300 min. In contrary, non-infected tissue showed a slightly increased trend of enzyme level up to 420 min. In infected tissue, a sharp transient increase (3.1 fold) of peroxidase activity appeared between 300 - 420 min post infections. Afterwards, significantly different but steady maintenance of enzyme levels were observed in two treatments. On the other hand, during symptom development, a sharp increase in peroxidase activity was noticed up to 4th grade of lesion appearance (25.1 % to 50% of leaf area infection), and then declined slightly. However, in non-infected but same age healthy leaves, such huge fluctuations of enzyme level did not apparent. A high positive correlation $(R^2=0.92)$ between peroxidase activity and leaf spot development grades was also marked. The result implies that pre-symptomatic burst (between 1 - 5 and 300 - 420 min) and subsequent increased trend of guaiacol peroxidase activity may require for the symptomatic manifestation of Myrothecium leaf spot in mulberry.