• Title/Summary/Keyword: Tissue specificity

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Characterization of tissue-specific mbu-3 gene expression in the mouse central nervous system

  • Lee, Chae-Jin;Cho, Eun-Young;Kim, Sun-Jung
    • BMB Reports
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    • v.41 no.12
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    • pp.875-880
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    • 2008
  • Mbu-3 is a novel mouse brain unigene that was identified by digital differential display. In this study, expression of the gene was chased through developmental stages and the protein product was identified in the brain. The cDNA sequence was 3,995-bp long and contained an ORF of 745 AA. Database searches revealed that the chicken SST273 gene containing LRR- and Ig-domain was an mbu-3 orthologue. Tissue specificity for the gene was examined in embryos and in brains at post-natal and adult stages. During the embryonic stages, mbu-3 was localized to the central nervous system in the brain and spinal cord. In the early post-natal stages, the gene was evenly expressed in the brain. However, with aging, expression was confined to specific regions, particularly the hippocampus. The protein was approximately 95 kDa as determined by Western blot analysis of brain extracts.

Gall structure and specificity in Bostrychia culture isolates (Rhodomelaceae, Rhodophyta)

  • West, John A.;Pueschel, Curt M.;Klochkova, Tatyana A.;Kim, Gwang Hoon;De Goer, Susan;Zuccarello, Giuseppe C.
    • ALGAE
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    • v.28 no.1
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    • pp.83-92
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    • 2013
  • The descriptions of galls, or tumors, in red algae have been sparse. K$\ddot{u}$tzing (1865) observed possible galls of Bostrychia but only presented a drawing. Intensive culture observations of hundreds of specimens of the genus Bostrychia over many years have revealed that galls appeared in only a small subset of our unialgal cultures of B. kelanensis, Bostrychia moritziana/radicans, B. radicosa, B. simpliciuscula, and B. tenella and continued to be produced intermittently or continuously over many years in some cultures but were never seen in field specimens. Galls appeared as unorganized tissue found primarily on males and bisexuals, but occasionally on females and tetrasporophytes. The gall cells usually were less pigmented than neighboring tissue, but contained cells with fluorescent plastids and nuclei. The galls were not transferable to other potential hosts. Galls could be produced from gall-free tissue of cultures that originally had galls even after transfer to new culture dishes. Electon microscopy of galls on one isolate (3895) showed that virus-like particles are observed in some gall cells. It is possible that a virus is the causative agent of these galls.

Improvement of Antigen Blotting in a Tissue Blot Immunobinding Assay for the Detection of Two Chili Pepper Viruses

  • Han, Jung-Heon;Shin, Jun-Sung;Kim, Young-Ho;Kim, Byung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.17 no.11
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    • pp.1885-1889
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    • 2007
  • The tissue blot immunobinding assay (TBIA) is widely used for the detection and localization of plant viruses in various plant tissues. The basic experimental procedures of TBIA sampling and blotting were simplified using commercially available micropipette tips. This method was termed the ring-blot immunobinding assay (R-BIA), as the blot on the membrane forms a ring shape. The detection efficacy of R-BIA was tested for two chili pepper viruses, pepper mild mottle tobamovirus (PMMoV) and pepper mottle potyvirus (PepMoV), following the optimized serological procedures of TBIA (length of the incubation period and BSA concentration, and primary and secondary antibodies). Sensitivity of the R-BIA was about 1 ng/ml of purified PMMoV in pepper leaf sap from a healthy pepper plant. R-BIA also showed high specificity in the detection of PMMoV and PepMoV. Moreover, the modified sampling and blotting procedures were simpler and more reliable than other TBIA methods (such as whole-leaf blotting and crushed-leaf blotting), suggesting that the R-BIA may be used for medium- to large-scale detection of plant viruses in laboratories with minimal facilities.

Enzyme-linked immunosorbent assay (ELISA) for the detection of RVS (Retrovirus of Salmonid) (ELISA법을 이용한 연어과 어류의 RVS 검출(Retrovirus of Salmonid) 검출)

  • Oh, Myung-Joo;Yoshimizu, Mamoru
    • Journal of fish pathology
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    • v.9 no.2
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    • pp.169-176
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    • 1996
  • An indirect double antibody enzyme-linked immunosorbent assay (ELISA) was developed for rapid detection of a new virus isolated from abnormally swimming salmonid fish, RVS (Retrovirus of salmonid). Results using brain tissue homogenates, and infected cell cultures are described. The sensitivity of the methods is $10^{2.6}$ $TCID_{50}/100{\mu}l$ of the examined cell culture fluid. The specificity was confirmed by the ELISA inhibition test and virological examinations. Viral antigen could be detected in artificially infected fish tissue homogenates. The assay will allow the diagnosis of RVS-infected fish within a day.

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Mechanism and Regulation of Amino Acid Transport in Mammary Gland - Review -

  • Kansal, Vinod K.;Sharma, Rekha
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.5
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    • pp.710-719
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    • 2001
  • Several amino acid transport systems in mammary gland have been characterized during the last few years. These systems may be divided into two broad categories based on whether they are sodium-dependent or $Na^{+}$-independent, and each of these categories is subdivided into 3 groups depending on whether the systems prefer zwitterionic, cationic or anionic substrates. The zwitterion preferring transport processes in mammary gland are $Na^{+}$-dependent system A and $Na^{+}$-independent systems L and T. System $y^{+}$ is a $Na^{+}$-independent transporter of cationic amino acids and $X_{AG^{-}}$ is a $Na^{+}$-dependent system for anionic amino acids. A ($Na^{+}+Cl^{-}$)-dependent system, selective for $\beta$-amino acids has been reported in rat mammary tissue. In addition, there is yet another class of transporters that have still broader specificity. The $Na^{+}$-dependent systems $BCl^{-}$-dependent and $BCl^{-}$-independent and $Na^{+}$-independent system $y^{+}L$ have been reported to mediate the transport of zwitterionic as well as cationic amino acids. Each system has been characterized with respect to its substrate specificity, affinity, kinetics and ion-dependence. Transport of amino acids by mammary tissue is regulated by i) the intracellular substrate concentration, ii) lactogenic hormones and iii) milk stasis. Four of the above transport systems (i.e. A, L, $y^{+}$ and $BCl^{-}$-independent) are up-regulated by lactogenic hormones (insulin, cortisol and prolactin) in mammary gland.

C-terminal Truncation Mutant of the Human ${\beta}_2$-adrenergic Receptor Expressed in E. coli as a Fusion Protein Retains Ligand Binding Affinity

  • Shin, Jin-Chul;Lee, Sang-Derk;Shin, Chan-Young;Lee, Sang-Bong;Ko, Kwang-Ho
    • Biomolecules & Therapeutics
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    • v.4 no.1
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    • pp.97-102
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    • 1996
  • To investigate whether human $\beta$$_2$-adrenergic receptor devoid of the C-terminal two transmembrane helices retain its ligand binding activity and specificity, 5'780-bp DNA fragment of the receptor gene which encodes amino acid 1-260 of human $\beta$$_2$-adrenergic receptor was subcloned into the bacterial fusion protein expression vector and expressed as a form of glutathione-S-transferase (GST) fusion protein in E. coli DH5$\alpha$. The receptor fusion protein was expressed as a membrane bound form which was verified by SDS-PAGE and Western blot. The fusion protein expressed in this study specifically bound $\beta$-adrenergic receptor ligand [$^3$H] Dihydroalprenolol. In saturation ligand binding assay, the $K_{d}$ value was 7.6 nM which was similar to that of intact $\beta$$_2$-adrenergic receptor in normal animal tissue ( $K_{d}$=1~2 nM) and the $B_{max}$ value was 266 fmol/mg membrane protein. In competition binding assay, the order of binding affinity of various adrenergic receptor agonists to the fusion protein was isoproterenol》epinephrine norepinephrine, which was similar to that of intact receptor in normal animal tissue. These results suggest that N-terminal five transmembrane helices of the $\beta$$_2$-adrenergic receptor be sufficient to determine the ligand binding activity and specificity, irrespective of the presence or absence of the C-terminal two transmembrane helices.s.s.s.

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Studies on the Development and Utilization of Polyclonal Antibodies Against Swine Adipocyte Plasma Membrane Proteins (돼지 지방세포 원형질막 단백질에 대한 다클론항체의 생산 및 이용에 관한 연구)

  • Baek, K.H.;Kwak, T.H.;Oh, Y.S.;Choi, C.W.;Jung, K.K.;Choi, Chang-Bon
    • Journal of Animal Science and Technology
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    • v.47 no.1
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    • pp.19-28
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    • 2005
  • The objectives of the current study were to develop polyclonal antibodies in sheep against adipocyte plasma membrane(APM) proteins isolated from swine, to investigate tissue specificity, and to determine cytotoxic effects of antiserum on swine adipocytes. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver, and spleen were isolated using a 32% sucrose gradient. Adult male sheep was immunized three times at three week interval with the purified swine APM proteins. Antiserum was taken from immunized sheep at 10, 12, and 14 days after the third immunization. Antiserum expressed strong reactivity with APM proteins determined by enzyme-linked immunosorbent assay(ELISA), and the reactivity could be detected at dilutions in excess of 1 : 81,000. Antiserum showed very low binding affinity with proteins isolated from brain, heart, kidney, liver, or spleen. Tissue specificity of the antiserum was reconfirmed by Western immunoblotting using anti-sheep immunoglobulin G•alkalinephosphatase conjugate as a secondary antibody. The reactivity of antiserum to the external surface of fixed swine adipocytes was confmned by an immunohistochemical technique using anti-sheep immunoglobulin G-FITC. Confluent swine adipocytes in culture were lysed by antiserum treatment and cytosolie lactate dehydrogenase(LDH) was released as a dose-dependent patterns while adipocytes treated with normal sheep serum maintained their integrity and expressed low level of LDH. These results implicate that fat contents in the pigs can be reduced by immunological methods.

Tissue Transglutaminase Antibody and Its Association with Duodenal Biopsy in Diagnosis of Pediatric Celiac Disease

  • Meena, Daleep K.;Akunuri, Shalini;Meena, Preetam;Bhramer, Ashok;Sharma, Shiv D.;Gupta, Rajkumar
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.22 no.4
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    • pp.350-357
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    • 2019
  • Purpose: This study aimed to evaluate a possible association between the anti-tissue transglutaminase antibody (anti-tTG) titer and stage of duodenal mucosal damage and assess a possible cut-off value of anti-tTG at which celiac disease (CD) may be diagnosed in children in conjunction with clinical judgment. Methods: This observational study was conducted at a gastroenterology clinic in a tertiary hospital from April 2012 to May 2013. Seventy children between 6-months and 18-years-old with suspected CD underwent celiac serology and duodenal biopsy. Statistical analyses were done using SPSS 16. Diagnostic test values were determined for comparing the anti-tTG titer with duodenal biopsy. An analysis of variance and Tukey-Kramer tests were performed for comparing the means between groups. A receiver operating characteristics curve was plotted to determine various cut-off values of anti-tTG. Results: The mean antibody titer increased with severity of Marsh staging (p<0.001). An immunoglobulin (Ig) A-tTG value at 115 AU/mL had 76% sensitivity and 100% specificity with a 100% positive predictive value (PPV) and 17% negative predictive value (NPV) for diagnosis of CD (p<0.001, 95% confidence interval [CI], 0.75-1). Conclusion: There is an association between the anti-tTG titer and stage of duodenal mucosal injury in children with CD. An anti-tTG value of 115 AU/mL (6.4 times the upper normal limit) had 76% sensitivity, 100% specificity, with a 100% PPV, and 17% NPV for diagnosing CD (95% CI, 0.75-1). This cut-off may be used in combination with clinical judgment to diagnose CD.

Diagnostic Value of Immunoglobulin G Anti-Deamidated Gliadin Peptide Antibody for Diagnosis of Pediatric Celiac Disease: A Study from Shiraz, Iran

  • Anbardar, Mohammad Hossein;Haghighi, Fatemeh Golbon;Honar, Naser;Zahmatkeshan, Mozhgan
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.25 no.4
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    • pp.312-320
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    • 2022
  • Purpose: Screening serologic tests are important tools for the diagnosis of celiac disease (CD). Immunoglobulin (Ig)G anti-deamidated gliadin peptide (anti-DGP) is a relatively new autoantibody thought to have good diagnostic accuracy, comparable to that of anti-tissue transglutaminase (anti-tTG) antibody. Methods: Pediatric patients (n=86) with a clinical suspicion of CD were included. Duodenal biopsy, anti-tTG, and IgG anti-DGP antibody tests were performed. The patients were divided into CD and control groups based on the pathological evaluation of duodenal biopsies. The diagnostic accuracy of serological tests was determined. Results: IgA anti-tTG and IgG anti-DGP antibodies were positive in 86.3% and 95.4% of patients, respectively. The sensitivity, specificity, and diagnostic accuracy of the IgA anti-tTG test were 86.3%, 50.0%, and 68.6%, respectively, and those of the IgG anti-DGP test were 95.4%, 85.7%, and 90.7%, respectively. The area under the receiver operating characteristic (ROC) curve was 0.84 (95% confidence interval [CI], 0.74-0.91) for IgA anti-tTG test and 0.93 (95% CI, 0.86-0.97) for IgG anti-DGP test. The comparison of IgA anti-tTG and IgG anti-DGP ROC curves showed a higher sensitivity and specificity of the IgG anti-DGP test. Conclusion: IgG anti-DGP is a reliable serological test for CD diagnosis in children. High tTG and DGP titers in the serum are suggestive of severe duodenal atrophy. The combined use of IgA anti-tTG and IgG anti-DGP tests for the initial screening of CD can improve diagnostic sensitivity.

Comparison of radiomics prediction models for lung metastases according to four semiautomatic segmentation methods in soft-tissue sarcomas of the extremities

  • Heesoon Sheen;Han-Back Shin;Jung Young Kim
    • Journal of the Korean Physical Society
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    • v.80
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    • pp.247-256
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    • 2022
  • Our objective was to investigate radiomics signatures and prediction models defined by four segmentation methods in using 2-[18F]fluoro-2-deoxy-d-glucose positron emission tomography (18F-FDG PET) imaging of lung metastases of soft-tissue sarcomas (STSs). For this purpose, three fixed threshold methods using the standardized uptake value (SUV) and gradient-based edge detection (ED) were used for tumor delineation on the PET images of STSs. The Dice coefficients (DCs) of the segmentation methods were compared. The least absolute shrinkage and selection operator (LASSO) regression and Spearman's rank, and Friedman's ANOVA test were used for selection and validation of radiomics features. The developed radiomics models were assessed using ROC (receiver operating characteristics) curve and confusion matrices. According to the results, the DC values showed the biggest difference between SUV40% and other segmentation methods (DC: 0.55 and 0.59). Grey-level run-length matrix_run-length nonuniformity (GLRLM_RLNU) was a common radiomics signature extracted by all segmentation methods. The multivariable logistic regression of ED showed the highest area under the ROC (receiver operating characteristic) curve (AUC), sensitivity, specificity, and accuracy (AUC: 0.88, sensitivity: 0.85, specificity: 0.74, accuracy: 0.81). In our research, the ED method was able to derive a significant model of radiomics. GLRLM_RLNU which was selected from all segmented methods as a meaningful feature was considered the obvious radiomics feature associated with the heterogeneity and the aggressiveness. Our results have apparently showed that radiomics signatures have the potential to uncover tumor characteristics.