• International Journal of Oral Biology
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• v.36 no.4
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• pp.167-172
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• 2011

LAR-RPTP (leukocyte common antigen-related receptor protein tyrosine phosphatase) is an important regulator in the nervous system, but little is known about its expression pattern in rat trigeminal ganglion (TG) neurons. To examine whether LAR-RPTP is expressed in the TG in the current study, we sacrificed rats at 0, 7, 10 and 56 day postpartum (dpp) and a second group of rats at 3 and 5 days after an experimental tooth extraction as a TG injury model. RT-PCR was then used to determine the level of LAR-RPTP expression in the TG and immunohistology was employed to detect the subcellular localization of the protein. The mRNA expression of LAR-RPTP during the developmental stages in the TG was found to gradually increase. After experimental tooth extraction however, these transcript levels had significantly decreased at three days. LAR-RPTP protein signals in the TG were found to be cytoplasmic in the normal animals but interestingly, at five days after an experimental tooth extraction, these signals were rare. These results indicate that LAR-RPTP may be regulated during both the developmental as well as regenerative processes that take place in the TG. This further suggests that LAR-RPTP is not only involved in primary axonogenesis but possibly also in the molecular control of axons during TG repair.

• BMB Reports
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• v.38 no.5
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• pp.571-576
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• 2005

Cytokines are proteins produced by many different cells of the immune system and play a significant role in initiating and regulating the inflammatory process. In this research, an important cytokine, interleukin-10 (IL-10) gene, has been identified and characterized from zebrafish (Danio rerio) genome database. Zebrafish IL-10 is located within a 2690 bp fragment and contains five exons and four introns, sharing the same organization with mammalian IL-10 genes. An open reading frame of 543 bp was found to encode a putative 180 amino acid protein with a signal peptide of 22 amino acids, which shares 29.7-80.9% homology with amino acid sequences of other known IL-10. The signature motif of IL-10 is also conserved in zebrafish IL-10. The predicted transcript was finally confirmed by sequencing of cDNA clones. Multi-tissue reverse transcriptase PCR (RT-PCR) was performed to examine the tissue distribution and expression regulation of this gene in seven organs of normal and lipopolysaccharide (LPS) stimulation zebrafish. The results demonstrated that this gene was expressed slightly in normal kidney, gill and gut, no expression was detected in other four tissues. The expression was clearly upregulated after LPS stimulation. Using the ideal zebrafish model, further study of IL-10 characterization and function may provide insight on the understanding of the innate immune system.

• Horticultural Science & Technology
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• v.30 no.6
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• pp.626-634
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• 2012

There are variations among grapevine genotypes in the levels of tolerance to cold, and cold-hardiness of grapevine has been affected by the change of short-term environment during over-wintering. In this study, the responses of vines to cold in open field and rain-shelter system were investigated to obtain useful information in increasing the tolerance to cold in grape cultivation. Total carbohydrate content of bearing mother branches was higher in the rain-shelter system than in the open field, and lower in the branches of 'Muscat Bailey A' than in 'Campbell Early' and 'Kyoho'. Bud-burst and shoot growth were better in the rain-shelter system than in open field, whereas there is no significant difference among the treatments of net beside vines. There was also low incidence of gray mold in rain shelter system. Stilbene compounds such as t-piceid, resveratrol, piceatannol, c-piceid were accumulated in the cold-treated shoot from vine cuts harvested in rain shelter system. Genes of chalcone isomerase, manganese superoxide dismutase, proline rich protein 2, and temperature induced lipocalin were highly expressed in the cold-treated shoot from vine cuts harvested in rain shelter system. While there was not change of air temperature, but high reduction of wind speed in the rain shelter system compared to open field, and increase in the reduction of wind speed by net treatment. The damage of grapevines by cold in the extreme low temperature could be reduced by keeping them in the rain shelter system with net during winter season.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.153-159
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• 2007

In this study, using FIV-based lentivirus vector system, we tried to express hG-CSF in tetracycline-controllable manner. hG-CSF influences the proliferation, differentiation, and survival of cells in the neutrophil lineage. To enhance stability and translation of hG-CSF transcript, WPRE sequence was also introduced into FIV-Tet-On vector at downstream region of either the hG-CSF gene or the sequence encoding rtTA. Primary culture cells (CEF, chicken embryonic fibroblast; PFF, procine fetal fibroblast) infected with the recombinant FIV were cultured in the medium supplemented with or without doxycycline for 48 hours, and induction efficiency was measured by comparing the hG-CSF gene expression level using quantitative real-time PCR, Western blot and ELISA. Higher hG-CSF expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hG-CSF (in CEF) or rtTA (in PEE) gene. This FIV-Tet-On vector system may be helpful in solving serious physiological disturbance problems which has continuously hampered successful production of transgenic animals and gene therapy.

• Biomolecules & Therapeutics
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• v.7 no.4
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• pp.307-314
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• 1999

The human muscarinic acetylcholine receptor (mAChR) subtypes Hml, Hm2 and Hm3 have been expressed in insect cells (Spodoptera frugiperda, Sf9) using the baculovirus expression system. Expression of relevant DNA, transcript and receptor proteins was identified by PCR, Northern blotting and [$^{3}H$]QNB binding, respectively. As assessed by [$^{3}H$]QNB binding sites, yields of muscarinic receptors in membrane preparations in this study were as about 5-20 times high as those in mammalian cells reported in previous studies. The [$^{3}H$]QNB competition binding studies with well-known subtype-selective mAChR antagonists showed that the receptors expressed in Sf9 cells retain the pharmacological characteristics expected for the ml , m2 and m3 muscarinic receptors. The ml-selective antagonist, pirenzepine, displayed a considerably higher affinity for Hml by 110-fold and 35-fold than for Hm2 and Hm3, respectively, The m2-selective methoctramine displayed a significantly higher affinity for Hm2 than for Hml and Hm3 (10- and 26-fold, respectively). p-F-HHSiD exhibited high affinity for Hm3 that is not significantly different from those for Hml, but 66-fold higher than its affinity for Hm2. The functional coupling of the recombinant receptors to second messenger systems was also examined. While both Hml and Hm3 stimulated phosphoinositide hydrolysis upon activation by carba-chol, Hm2 produced no response. On the other hand, activation of mAChRs induced the inhibition of forsko-lin-stimulated cyclic AMP formation in Hm2-expressing cells, whereas the significant dose-dependent increase in or poor response on cyclic AMP formation were produced in Hml or Hm3-expressing cells, respectively. These results indicate the differential coupling of recombinant Hml, Hm2 and Hm3 receptors expressed in SF9 cells to intracellular signalling system.

• Korean Journal of Social Welfare
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• v.55
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• pp.231-254
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• 2003

The main premise of this study is that the assurance on human rights of the mentally ill is contradictory to 'the greatest happiness of the greatest number' or 'therapeutic benefits'. Accordingly this study aims to find out the conceptual framework of assurance and restriction on human rights of the mentally ill. Using qualitative method, this study conducted the in-depth interview with 10 general public, 9 professionals, 6 mentally ill regarding the several issues of human rights included long-term hospitalization, restriction on driver's licence, involuntary hospitalization, restriction on communication in psychiatric ward. Research results are as follows ; two sets of dimensions are inferred from the analysis of interview transcript. First dimension is the focus of justification, ranging from the emphasis on positive consequences influenced to 'the greatest numbers'(the utilitarian thought) to the emphasis on assurance of rights without any conditions(the deontological thought). Second dimension is the locus of decision, raging from the formal system included the professionals and the government agency to the informal system included the families and the mentally ill. And there are differences in attitudes toward assurance and restriction on rights of the mentally ill among general public, professionals, and the mentally ill. In detail, general public regarded that 'involuntary hospitalization' and 'restriction on driver's license' which are apt to be directly harmful to people must be justified by consequence-centered and decided by the formal system, while 'long-term hospitalization' which is less harmful to people could be justified by right-centered and decided by the formal-system. And they thought that 'restriction of communication' could be justified by right-centered and decided by the informal system. Based on the findings, this study would conclude that practical guidelines for the promotion of human rights of the mentally ill must be developed.

• Tuberculosis and Respiratory Diseases
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• v.79 no.2
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• pp.85-90
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• 2016

Background: Lung cancer is the most common cause of cancer related death. Alterations in gene sequence, structure, and expression have an important role in the pathogenesis of lung cancer. Fusion genes and alternative splicing of cancer-related genes have the potential to be oncogenic. In the current study, we performed RNA-sequencing (RNA-seq) to investigate potential fusion genes and alternative splicing in non-small cell lung cancer. Methods: RNA was isolated from lung tissues obtained from 86 subjects with lung cancer. The RNA samples from lung cancer and normal tissues were processed with RNA-seq using the HiSeq 2000 system. Fusion genes were evaluated using Defuse and ChimeraScan. Candidate fusion transcripts were validated by Sanger sequencing. Alternative splicing was analyzed using multivariate analysis of transcript sequencing and validated using quantitative real time polymerase chain reaction. Results: RNA-seq data identified oncogenic fusion genes EML4-ALK and SLC34A2-ROS1 in three of 86 normal-cancer paired samples. Nine distinct fusion transcripts were selected using DeFuse and ChimeraScan; of which, four fusion transcripts were validated by Sanger sequencing. In 33 squamous cell carcinoma, 29 tumor specific skipped exon events and six mutually exclusive exon events were identified. ITGB4 and PYCR1 were top genes that showed significant tumor specific splice variants. Conclusion: In conclusion, RNA-seq data identified novel potential fusion transcripts and splice variants. Further evaluation of their functional significance in the pathogenesis of lung cancer is required.

• The Plant Pathology Journal
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• v.30 no.4
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• pp.355-366
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• 2014

In this study, resistance responses were investigated during the interaction of Botrytis fabae with two faba bean cultivars expressing different levels of resistance against this pathogen, Nubaria (resistant) and Giza 40 (susceptible). Disease severity was assessed on leaves using a rating scale from 1 to 9. Accumulation levels of reactive oxygen species (ROS), lipid peroxidation and antioxidant enzymes (superoxide dismutase, catalase and ascorbate peroxidase) were measured in leaf tissues at different times of infection. The expression profiles of two pathogenesis-related proteins (PRPs) encoded by the genes PR-1 and ${\beta}$-1,3-glucanase were also investigated using reverse transcription RT-PCR analysis. The accumulation of these defense responses was induced significantly in both cultivars upon infection with B. fabae compared with un-inoculated controls. The resistant cultivar showed weaker necrotic symptom expression, less ROS accumulation, a lower rate of lipid peroxidation and higher activity of the enzymatic ROS scavenging system compared with susceptible cultivar. Interestingly, ROS accumulated rapidly in the resistant leaf tissues and peaked during the early stages of infection, whereas accumulation was stronger and more intense in the susceptible tissues in later stages. Moreover, the response of the resistant cultivar to infection was earlier and stronger, exhibiting high transcript accumulation of the PR genes. These results indicated that the induction of oxidant/antioxidant responses and the accumulation of PRPs are part of the faba bean defense mechanism against the necrotrophic fungus B. fabae with a different intensity and timing of induction, depending on the resistance levels.

• Journal of Microbiology and Biotechnology
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• v.24 no.8
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• pp.1065-1072
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• 2014

Doxorubicin, produced by Streptomyces peucetius ATCC 27952, is tightly regulated by dnrO, dnrN, and dnrI regulators. Genome mining of S. peucetius revealed the presence of the IclR (doxR) type family of transcription regulator mediating the signal-dependent expression of operons at the nonribosomal peptide synthetase gene cluster. Overexpression of doxR in native strain strongly repressed the drug production. Furthermore, it also had a negative effect on the regulatory system of doxorubicin, wherein the transcript of dnrI was reduced to the maximum level in comparision with the other two. Interestingly, the overexpression of the same gene also had strong inhibitory effects on the production of actinorhodin (blue pigment) and undecylprodigiosin (red pigment) in Streptomyces coelicolor M145, herboxidiene production in Streptomyces chromofuscus ATCC 49982, and spinosyn production in Saccharopolyspora spinosa NRRL 18395, respectively. Moreover, DoxR exhibited pleiotropic effects on the production of blue and red pigments in S. coelicolor when grown in different agar media, wherein the production of blue pigment was inhibited in R2YE medium and the red pigment was inhibited in YEME medium. However, the production of both blue and red pigments from S. coelicolor harboring doxR was halted in ISP2 medium, whereas S. coelicolor produced both pigmented antibiotics in the same plate. These consequences demonstrate that the on and off production of these antibiotics was not due to salt stress or media compositions, but was selectively controlled in actinomycetes.

• Journal of Microbiology and Biotechnology
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• v.23 no.5
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• pp.587-601
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• 2013

During the last few decades, it has become evident that the compatibility of the yeast biochemical environment with the ability to process and translate the RNA transcript, along with its capacity to modify a translated protein, are relevant requirements for selecting this host cell for protein expression in several pharmaceutical and clinical applications. In particular, Pichia pastoris is used as an industrial host for recombinant protein and metabolite production, showing a powerful capacity to meet required biomolecular target production levels in high-throughput assays for functional genomics and drug screening. In addition, there is a great advantage to using P. pastoris for protein secretion, even at high molecular weights, since the recovery and purification steps are simplified owing to relatively low levels of endogenous proteins in the extracellular medium. Clearly, no single microexpression system can provide all of the desired properties for human protein production. Moreover, chemical and physical bioprocess parameters, including culture medium formulation, temperature, pH, agitation, aeration rates, induction, and feeding strategies, can highly influence product yield and quality. In order to benefit from the currently available wide range of biosynthesis strategies using P. pastoris, this mini review focuses on the developments and technological fermentation achievements, providing both a comparative and an overall integration analysis. The main aim is to highlight the relevance and versatility of the P. pastoris biosystem to the design of more cost-effective microfactories to meet the increasing demands for recombinant membrane proteins and clinical antibodies for several therapeutic applications.