• Title/Summary/Keyword: Histochemical analysis

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Clinical Features and Diagnosis of Hirschsprung's Disease (Hirschsprung's Disease의 임상 양상 및 진단)

  • Park, Woo-Hyun
    • Advances in pediatric surgery
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    • v.8 no.1
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    • pp.48-53
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    • 2002
  • Diagnosing Hirschprungs disease (HD) is a clinical challenge to pediatric surgeons. The cardinal symptoms are failure of passage of meconium within first 24 hours of life, abdominal distension, and vomiting. The severity of these symptoms and the degree of consitpation vary considerably between patients. HD is suspected on the basis of history and clinical findings and the diagnosis is established by radiological examination, anorectal manometry, and histochemical analysis of biopsy specimens. In this review, the advantages and pitfalls of each diagnostic modality are discussed. And a diagnostic approach utilizing these diagnostic modalities in children with suspicious HD is presented.

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Expression and Inheritance Patterns of Gus Gene Driven by an Endosperm-Specific Promoter in Transgenic Tobacco (배유 특이 프로모터에 의해 유도된 GUS 유전자의 형질전환 담배 내에서의 발현 및 유전 양상)

  • Park, Young Doo;Kim, Hyoung Seok
    • Horticultural Science & Technology
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    • v.18 no.5
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    • pp.594-598
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    • 2000
  • This study was carried out to investigate the tissue-specific expression of ${\beta}$-glucuronidase (gus) gene driven by endosperm-specific promoter (Z4 promoter) in the transgenic tobacco and to find out inheritance pattern of transgene to the next generation. Tobacco (Nicotiana tabaccum cv. Havana SR1) was transformed with Agrobacterium tumerfaciens LBA4404 harboring BV3 construct containing gus gene driven by Z4 promoter and a kanamycin resistant gene. Seven hundred bp PCR products, indicating the presence of npt II gene, were found in the all eight transformants by PCR analysis using nptII primers. To study the expression pattern of the two different kind of promoters, leaf disks of the Z4pro-gus-transformed plants and 35Spro-gus-transformed plants were analyzed histochemically for gus activity. As a result, leaf disks of Z4pro-gus-transformed plants showed very weak and partial positive gus activity. In contrast, leaf disks of 35Spro-gus-transformed plants showed relatively strong positive gus activity. To investigate the expressed position of Z4 promoter, seeds from Z4pro-gus-transformed plants and 35Spro-gus-transformed plants were analyzed histochemically for gus activity. Z4pro-gus-transformed seeds showed positive gus activity restricted to the endosperm. However, the blue-colored product in 35Spro-gus-transformed seeds was observed in all the area including endosperm. Kanamycin resistance assay showed that transgenes were stably inherited to next generation in all lines.

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Comparative Analysis of Histochemical Stains about Detection of H. pylori in Gastric Mucosa (위점막에서 H. pylori 검출을 위한 조직염색법의 비교)

  • Cha, Min-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.3
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    • pp.223-230
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    • 2007
  • Several methods have been used in the detection of Helicobacter pylori (H. pylori) which was believed to be a pathogenic organism causing chronic gastritis, benign peptic ulcer, gastric carcinoma or malignant lymphoma. Even though several methods were introduced for detection of H. pylori in stomach, there were controversies in their sensitivities and specificities. This experiment were designed to study the comparative analysis of staining methods (hematoxylin and eosin (H&E), Giemsa, Warthin-Starry and immunohistochemical stain) to dectect H. pylori in the gastric mucosa. The results were as follows. Average density score of H. pylori classified by Genta were 2.29 in Warthin-Starry stain, 2.19 in Giemsa stain, 1.34 in immunohistochemical stain and 0.98 in H&E stain. By comparison between inflammatory degree by Sydney system and result of Warthin-Starry stain, the detection rate and densities of H. pylori were increased from mild (61.5% and 0.8), moderate (90.4 and 2.1), and severe (100% and 3.2). From the above findings, Warthin-Starry stain is useful method for detection of H. pylori in gastric mucosa.

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Isolation of Multi-Abiotic Stress Response Genes to Generate Global Warming Defense Forage Crops

  • Ermawati, Netty;Hong, Jong Chan;Son, Daeyoung;Cha, Joon-Yung
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.41 no.4
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    • pp.242-249
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    • 2021
  • Forage crop management is severely challenged by global warming-induced climate changes representing diverse a/biotic stresses. Thus, screening of valuable genetic resources would be applied to develop stress-tolerant forage crops. We isolated two NAC (NAM, ATAF1, ATAF2, CUC2) transcription factors (ANAC032 and ANAC083) transcriptionally activated by multi-abiotic stresses (salt, drought, and cold stresses) from Arabidopsis by microarray analysis. The NAC family is one of the most prominent transcription factor families in plants and functions in various biological processes. The enhanced expressions of two ANACs by multi-abiotic stresses were validated by quantitative RT-PCR analysis. We also confirmed that both ANACs were localized in the nucleus, suggesting that ANAC032 and ANAC083 act as transcription factors to regulate the expression of downstream target genes. Promoter activities of ANAC032 and ANAC083 through histochemical GUS staining again suggested that various abiotic stresses strongly drive both ANACs expressions. Our data suggest that ANAC032 and ANAC083 would be valuable genetic candidates for breeding multi-abiotic stress-tolerant forage crops via the genetic modification of a single gene.

Schizandrin, Oil Compounds, and Their Extraction Yield in Fruits of Schizandra chinensis Baillon (오미자의 Schizandrin과 기름성분 및 추출수율 변화)

  • 김관수;박춘근;류수노;방진기;이봉호
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.3
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    • pp.158-162
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    • 2000
  • Histochemical distribution and varietal variation of schizandrin and oil compounds, and the changes of their extraction yield were investigated in fruits of collected Schizandra chinensis including Inje cultivar. In histochemical analysis on the distribution of schizandrin and oil in fruits of Inje cultivar, higher concentrations of them were found in the seed (1.01% and 27.6%, respectively) than in the epicarp and mesocarp of the whole fruit. Average contents of schizandrin in fruits and oil in seeds of collected lines were 0.84% and 27.9%, respectively. The mean composition of fatty acids in seeds oil was 3.6% of palmitic acid, 0.6% of stearic acid, 19.7% of oleic acid, 73.0% of linoleic acid, and 3.1% of linolenic acid, showing high composition(95.8%) of total unsaturated fatty acid. Oil extracted from seeds of Inje cultivar contained 4.29% of schizandrin, indicating that seed oil contained much schizandrin, a bioactive lipid-soluble compound. Compared with 80% methanol extraction in fruits and seeds, yields of schizandrin and oil were lower showing 23.8% and 17.3%, respectively in boiling water extraction of the fruits and seeds without grinding. The seeds soaked with water during four months contained 1.18% of schizandrin and 25.2% of oil, whose contents were similar to those of the seeds stored at room temperature. These results demonstrated that the seed in the whole fruit could be utilized as a source to extract its functional oil and bioactive lipid-soluble compounds like schizandrin, especially after using Schizandra fruits for the beverage manufacture.

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Histochemical Analysis of the Cutaneous Wound Healing in the Amphibian (양서류 피부 상처회복과정에 대한 조직화학적 분석)

  • Lim, Do-Sun;Jeong, Soon-Jeong;Jeong, Je-O;Park, Joo-Cheol;Kim, Heung-Joong;Moon, Myung-Jin;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.34 no.1
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    • pp.1-11
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    • 2004
  • The wound healing is very complex biological processing including inflammatory, reepithelialization and matrix construction. According to the biological systematic category, the ability of the healing is very different. Generally healing ability of the lower animal group has been known more excellent compared to its higher group. Therefore, lower animals have been used as the experimental model to explore the mechanism of the wound healing or repair. To verify histochemical characteristics of the wound healing, we have used skin of the frog (Bombina orientalis) as known common amphibian. At day 1, 10, and 16, the mucous substance was very actively synthesized and strong positive by PAS and Alcian blue (pH 2.5). Day 10 after wounding, margin of the wound was gradually strong positive by PTAH staining for detection of collagen synthesis. At 3 to 6 hour and day 23 to 27, we have found the cell division was active through the MG-P staining, in which the concentration and division of DNA in nucleus was green to deep blue color.

Intragenic Control of Expression of a Rice MADS Box Gene OsMADS1

  • Jeon, Jong-Seong;Lee, Sichul;An, Gynheung
    • Molecules and Cells
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    • v.26 no.5
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    • pp.474-480
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    • 2008
  • OsMADS1 is a rice MADS box gene necessary for floral development. To identify the key cis-regulatory regions for its expression, we utilized transgenic rice plants expressing GUS fusion constructs. Histochemical analysis revealed that the 5.7-kb OsMADS1 intragenic sequences, encompassing exon 1, intron 1, and a part of exon 2, together with the 1.9-kb 5' upstream promoter region, are required for the GUS expression pattern that coincides with flower-preferential expression of OsMADS1. In contrast, the 5' upstream promoter sequence lacking this intragenic region caused ectopic expression of the reporter gene in both vegetative and reproductive tissues. Notably, incorporation of the intragenic region into the CaMV35S promoter directed the GUS expression pattern similar to that of the endogenous spatial expression of OsMADS1 in flowers. In addition, our transient gene expression assay revealed that the large first intron following the CaMV35S minimal promoter enhances flower-preferential expression of GUS. These results suggest that the OsMADS1 intragenic sequence, largely intron 1, contains a key regulatory region(s) essential for expression.

The Relationships between Muscle Fiber Characteristics, Intramuscular Fat Content, and Fatty Acid Compositions in M. longissimus lumborum of Hanwoo Steers

  • Joo, Seon-Tea;Joo, Sung-Hyun;Hwang, Young-Hwa
    • Food Science of Animal Resources
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    • v.37 no.5
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    • pp.780-786
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    • 2017
  • The objective of this study was to investigate the relationship between muscle fiber characteristics, intramuscular fat (IMF) content, and fatty acids composition in longissimus lumborum (LL) muscle from Hanwoo steers. The LL muscles were obtained from four quality grades (QG) carcasses and subjected to histochemical analysis. There were significant (p<0.05) differences in fiber number percentage (FNP) and fiber area percentage (FAP) of muscle fiber types among muscles from four QGs. Both FNP and FAP of type I increased while those of type IIB decreased with increasing QG from QG 2 to QG $1^{{+}{+}}$ (p<0.05). Also, with increasing QG, the saturated fatty acid (SFA) proportion decreased while monounsaturated fatty acid (MUFA) increased significantly (p<0.05). IMF content was positively correlated with both FNP and FAP of type I, but negatively correlated with those of type IIB. The proportions of SFA and MUFA were significantly (p<0.001) correlated with both type I and IIB composition. These results implied that muscle fiber type composition is an important factor influencing fatty acid composition in LL muscle of Hanwoo steer.

Efficient transformation of Actinidia arguta by reducing the strength of basal salts in the medium to alleviate callus browning

  • Han, Meili;Gleave, Andrew P.;Wang, Tianchi
    • Plant Biotechnology Reports
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    • v.4 no.2
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    • pp.129-138
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    • 2010
  • An efficient transformation system for high-throughput functional genomic studies of kiwifruit has been developed to overcome the problem of necrosis in Actinidia arguta explants. The system uses Agrobacterium tumefaciens strain EHA105 harbouring the binary vector pART27-10 to inoculate leaf strips. The vector contains neomycin phosphotransferase (nptII) and ${\beta}$-glucuronidase (GUS) (uidA) genes. A range of light intensities and different strengths of Murashige and Skoog (MS) basal salt media was used to overcome the problem of browning and/or necrosis of explants and calli. Callus browning was significantly reduced, resulting in regenerated adventitious shoots when the MS basal salt concentration in the culture medium was reduced to half-strength at low light intensity ($3.4\;{\mu}mol\;m^{-2}\;s^{-1}$) conditions. Inoculated leaf strips produced putative transformed shoots of Actinidia arguta on half-MS basal salt medium supplemented with 3.0 $mg\;l^{-1}$ zeatin, 0.5 $mg\;l^{-1}$ 6-benzyladenine, 0.05 $mg\;l^{-1}$ naphthalene acetic acid, 150 $mg\;l^{-1}$ kanamycin and 300 $mg\;l^{-1}$ $Timentin^{(R)}$. All regenerated plantlets were deemed putativ transgenic by histochemical GUS assay and polymerase chain-reaction analysis.

Structure and Expression of a Perilla (Perilla frutescens Britt) Gene, PfFAD3, Encoding the Microsomal ${\omega}-3$ Fatty Acid Desaturase

  • Lee, Hyang-Hwa;Pyee, Jae-Ho
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.5
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    • pp.424-427
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    • 2004
  • Microsomal ${\omega}-3$ fatty acid desaturase (FAD3) is an essential enzyme in the production of the n-3 polyunsaturated fatty acid ${\alpha}-linolenic$ acid during the seed developing stage. To understand the regulatory mechanism of the gene encoding the ${\omega}-3$ fatty acid desaturase, a genomic fragment corresponding to the previously isolated perilla seed PfFAD3 cDNA was amplified from perilla (Perilla frutescens Britt) by GenomeWalker PCR. Sequence analysis of the fragment provided with identification of a 1485-bp 5'-upstream region and a 241-bp intron in the open reading frame. To determine the tissue-specificity of the PfFAD3 gene expression, the 5'-upstream region was fused to the ${\beta}-glucuronidase$ (GUS) gene and incorporated into Arabidopsis thaliana. Histochemical assay of the transgenic plants showed that GUS expression was restricted to seed and pollen, showing that PfFAD3 gene was exclusively expressed in those tissues.