Background : Phospholipase C(PLC) plays an important role in cellular signal transduction and is thought to be critical in cellular growth, differentiation and transformation of certain malignancies. Two second messengers produced from the enzymatic action of PLC are diacylglycerol (DAG) and inositol 1, 4, 5-trisphosphate (IP3). These two second messengers are important in down stream signal activation of protein kinase C and intracellular calcium elevation. In addition, functional domains of the PLC isozymes, such as Src homology 2 (SH2) domain, Src homology 3 (SH3) domain, and pleckstrin homology (PH) domain play crucial roles in protein translocation, lipid membrane modificailon and intracellular memrane trafficking which occur during various mitogenic processes. We have previously reported the presence of PLC-${\gamma}1$, ${\gamma}2$, ${\beta}1$, ${\beta}3$, and ${\delta}1$ isozymes in normal human lung tissue and tyrosine-kinase-independent activation of phospholipase C-${\gamma}$ isozymes by tau protein and AHNAK. We had also found that the expression of AHNAK protein was markedly increased in various mstologic types of lung can∞r tissues as compared to the normallungs. However, the report concerning expression of various PLC isozymes in lung canærs and other lung diseases is lacking. Therefore, in this study we examined the expression of PLC isozymes in the paired surgical specimens taken from lung cancer patients. Methods : Surgically resected lung cancer tissue samples taken from thirty seven patients and their paired normal control lungs from the same patients, The expression of various PLC isozymes were studied. Western blot analysis of the tissue extracts for the PLC isozymes and immunohistochemistry was performed on typical samples for localization of the isozyme. Results : In 16 of 18 squamous cell carcinomas, the expression of PLC-${\gamma}1$ was increased. PLC-${\gamma}1$ was also found to be increased in all of 15 adenocarcinoma patients. In most of the non-small cell lung cancer tissues we had examined, expression of PLC-${\delta}1$ was decreased. However, the expression of PLC-${\delta}1$ was markedly increased in 3 adenocarcinomas and 3 squamous carcinomas. Although the numbers were small, in all 4 cases of small cell lung cancer tissues, the expression of PLC-${\delta}1$ was nearly absent. Conclusion : We found increased expression of PLC-${\gamma}1$ isozyme in lung cancer tissues. Results of this study, taken together with our earlier findings of AHNAK protein-a putative PLD-${\gamma}$, activator-over-expression, and the changes observed in PLC-${\delta}1$ in primary human lung cancers may provide a possible insight into the derranged calcium-inositol signaling pathways leading to the lung malignancies.
Journal of the Korea Academia-Industrial cooperation Society
/
v.19
no.10
/
pp.578-589
/
2018
Building Integrated Photovoltaic (BIPV) system is a typical integrated system that simultaneously performs both building function and solar power generation function. To maximize its potential advantage, however, the solar photovoltaic power generation function must be integrated from the early conceptual design stage, and maximum power generation must be designed. To cope with such requirements, preliminary research on BIPV design process based on architectural design model and computer simulation results for improving solar power generation performance have been published. However, the requirements of the BIPV system have not been clearly identified and systematically reflected in the subsequent design. Moreover, no model has verified the power generation design. To solve these problems, we systematically model the requirements of BIPV system and study power generation design based on the system requirements model. Through the study, we consistently use the standard system modeling language, SysML. Specifically, stakeholder requirements were first identified from stakeholders and related BIPV standards. Then, based on the domain model, the design requirements of the BIPV system were derived at the system level, and the functional and physical architectures of the target system were created based on the system requirements. Finally, the power generation performance of the BIPV system was evaluated through a simulated SysML model (Parametric diagram). If the SysML system model developed herein can be reinforced by reflecting the conditions resulting from building design, it will open an opportunity to study and optimize the power generation in the BIPV system in an integrated fashion.
Park, Jun Eun;Choi, Kang Duk;Kim, Sung Hwan;Hahm, Dae-Hyun;Seo, Jong Jin
Clinical and Experimental Pediatrics
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v.48
no.7
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pp.779-788
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2005
Purpose : Echinacea, a traditional plant medicine has been used as immune-stimulant. Recent studies have revealed that extract of Echinacea has immunostimulatory effects on human blood mononuclear cells. This study was designed for the purpose of screening the genes associated with immunologic effects of Echinacea on monocytes and dendritic cells using a cDNA microarray chip. Methods : $CD14^+$ monocyte cells were cultured for one day with Echinacea extract(final concentration : $50{\mu}g/mL$) in experiment 1, but were cultured without Echinacea in experiment 2. The gene expression of these cultured monocytes was analyzed using the cDNA microarray chip. Dendritic cells produced from $CD14^+$ monocyte were cultured for five days with GM-CSF and IL-4, and then cultured for one day with Echinacea in experiment 3, but were done without Echinacea in experiment 4. Results : In experiments 1 and 2, there were 17 significantly expressed genes with average expression ratios above 2.5, including interferon gamma-inducible protein 30(IFI 30), CDC(cell-division-cylcle)-like kinase 2(CLK 2), syndecan binding protein(syntenin), superoxide dismutase 2, etc. In experiments 3 and 4, there were 24 gene, with significantly expressed genes were 24 genes, which were insulin-like growth factor 2(somatomedin A), methyl-CpG binding domain protein 3, IFI 30, small inducible cytokine subfamily A, member 22, etc. The genes encoding CD44, IFI 30, mannose receptor C type 1(MRC 1), chemokine receptor 7(CCR 7), CLK 2, syntenin and cytochrome C oxidase subunit VIII were significantly expressed in both monocytes and dendritic cells cultured with Echinacea. Conclusion : This study employed a cDNA microarray chip to elicit the immune-associated gene profile; the expression was enhanced by Echinacea in CD14+ monocytes and dendritic cells. Thus we laid the basis for the quantitative and functional analysis of genes induced by Echinacea in monocytes and monocyte-derived dendritic cells.
Enzyme immunoassay to analyze specific binding activity of antibody to antigen uses horseradish peroxidase (HRP) or alkaline phosphatase (AP). Chemical methods are usually used for coupling of these enzymes to antibody, which is complicated and random cross-linking process. As results, it causes decreases or loss of functional activity of either antibody or enzyme. In addition, most enzyme assays use secondary antibody to detect antigen binding activity of primary antibody. Enzymes coupled to secondary antibody provide a binding signal by substrate-based color development, suggesting secondary antibody is required in enzyme immunoassay. Additional incubation time for binding of secondary antibody should also be necessary. More importantly, non-specific binding activity caused by secondary antibody should also be eliminated. In this study, we cloned AP isolated from Escherichia coli (E. coli) chromosome by PCR and fused to) hAY4 single-chain variable domain fragment (ScFv) specific to death receptor (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand (TRAIL). hAY4 ScFv-AP expressed in E. coli showed 73.8 kDa as a monomer in SDS-PAGE. However, this fusion protein shown in size-exclusion chromatography (SEC) exhibited 147.6 kDa as a dimer confirming that natural dimerization of AP by non-covalent association induced ScFv-AP dimerization. In several immunoassay such as ELISA, Western blot and immunocytochemistry, it showed antigen binding activity by color development of substrates catalyzed by AP directly fused to primary hAY4 ScFv without secondary antibody. In summary, hAY4 ScFv-AP fusion protein was successfully purified as a soluble dimeric form in E. coli and showed antigen binding activity in several immunoassays without addition of secondary antibody which sometimes causes time-consuming, expensive and non-specific false binding.
Rice seed maturation and germination involve drastic changes in water and nutrient transport, in which tonoplast aquaporins may play an important role. In the present study, gene expression profiles of 10 tonoplast intrinsic proteins (TIP) from rice were investigated by RT-PCR during seed development and germination. OsTIP3;1 and OsTIP3;2 were specifically expressed in mature seeds. Their transcript level rapidly decreased after onset of seed germination and gene expression was induced by ABA treatment. In contrast, expression of OsTIP2;1 and OsTIP4;3 was not seed specific as transcripts were found in vegetative tissues as well. Their respective transcript levels decreased at an early stage of seed development, whereas they increased at a later stage of seed germination and elongation of embryonic roots and shoots. When seed germination was inhibited by various stress conditions and ABA, expression of OsTIP2;1 and OsTIP4;3 was completely suppressed. In contrast, the expression level of OsTIP2;2 rapidly increased after seed imbibition and the transcript level was maintained under conditions inhibiting seed germination. These results implicate that tissue specific and developmental transcriptional regulation of OsTIPs in rice seeds depends on their specific function. In addition, OsTIPs can be discriminated by different potential phosphorylation and methylation sites in their protein structures. OsTIP3;1 and OsTIP3;2 possess unique phosphorylation signatures at their N-terminal domain, loop B and loop E, respectively. OsTIP2;1 and OsTIP4;3 have a potential methylation site at their Nterminal domain. This suggests that activity of specific tonoplast aquaporins may be regulated by post-translational modification as well as by transcriptional control.
Purpose: Spirituality is an important domain and is related with physical and psychological symptoms in terminal cancer patient. The aim of this study is to examine how patients' spirituality is associated with their physical and psychological symptoms as it has been explored by few studies. Methods: In this cross sectional study, 50 patients in the palliative ward of a tertiary hospital were interviewed. Spiritual well-being, depression, anxiety and pain is measured by Functional Assessment of Chronic-Illness Therapy-Spirituality (FACIT-Sp), hospital anxiety and depression scale (HADS) and the Korean version of the Brief Pain Inventory (BPI-K). The correlations between patients' spiritual well-being and anxiety, depression and pain were analysed. The association between spiritual well-being and age, gender, palliative performance scale (PPS), religion, mean pain intensity, anxiety, depression were assessed by univariate and multivariate regression analyses. Results: Spiritual well-being was negatively correlated with the mean pain intensity (r=-0.283, P<0.05), anxiety (r=-0.613, P<0.05) and depression (r=-0.526, P<0.05). In multivariate regression analysis, spiritual well-being showed negative association with anxiety (OR=-1.03, 95% CI=-1.657~-0.403, P=0.002) and positive association with the existence of religion (OR=9.193, 95% CI=4.158~14.229, P<0.001). Conclusion: In this study, patients' anxiety and existence of religion were significantly associated with spiritual well-being after adjusting age, gender, PPS, mean pain intensity, depression. Prospective studies are warranted.
Peroxiredoxins (Prxs) are ubiquitously distributed and play important functions in diverse cellular signaling systems. The proteins are largely classified into three groups, such as typical 2-Cys Prx, atypical 2-Cys Prx, and 1-Cys Prx, that are distinguished by their catalytic mechanisms and number of Cys residues. From the three classes of Prxs, the typical 2-Cys Prx containing the two-conserved Cys residues at its N-terminus and C-terminus catalyzes $H_2O_2$ with the use of thioredoxin (Trx) as an electron donor. During the catalytic cycle, the N-terminal Cys residue undergoes a peroxide-dependent oxidation to sulfenic acid, which can be further oxidized to sulfinic acid at the presence of high concentrations of $H_2O_2$ and a Trx system containing Trx, Trx reductase, and NADPH. The sulfinic acid form of 2-Cys Prx is reduced by the action of sulfiredoxin which requires ATP as an energy source. Under the strong oxidative or heat shock stress conditions, 2-Cys Prx in eukaryotes rapidly switches its protein structure from low-molecular-weight species to high-molecular-weight protein structures. In accordance with its structural changes, the protein concomitantly triggers functional switching from a peroxidase to a molecular chaperone, which can protect its substrate denaturation from external stress. In addition to its N-terminal active site, the C-terminal domain including 'YF-motif' of 2-Cys Prx plays a critical role in the structural changes. Therefore, the C-terminal truncated 2-Cys Prxs are not able to regulate their protein structures and highly resistant to $H_2O_2$-dependent hyperoxidation, suggesting that the reaction is guided by the peroxidatic Cys residue. Based on the results, it may be concluded that the peroxidatic Cys of 2-Cys Prx acts as an '$H_2O_2$-sensor' in the cells. The oxidative stress-dependent regulation of 2-Cys Prx provides a means of defense systems in cells to adapt stress conditions by activating intracellular defense signaling pathways. Particularly, 2-Cys Prxs in plants are localized in chloroplasts with a dynamic protein structure. The protein undergoes conformational changes again oxidative stress. Depending on a redox-potential of the chloroplasts, the plant 2-Cys Prx forms super-molecular weight protein structures, which attach to the thylakoid membranes in a reversible manner.
To date, we have considered the theoretical views, the standpoint of states and the discourse within the international community such as the UN Committee on the Peaceful Uses of Outer Space(COPUOS) regarding the Air/Outer Space Boundary Question which is one of the first issues of UN COPUOS established in line with marking the starting point of Outer Space Area. As above mentioned, discussions in the United Nations and among scholars of within each state regarding the delimitation issue often saw a division between those in favor of a functional approach (the functionalists) and those seeking the delineation of a boundary (the spatialists). The spatialists emphasize that the boundary between air and outer space should be delimited because the status of outer space is a type of public domain from which sovereign jurisdiction is excluded, as stated in Article 2 of Outer Space Treaty. On the contrary art. I of Chicago Convention is evidence of the acknowledgement of sovereignty over airspace existing as an international customary law, has the binding force of which exists independently of the Convention. The functionalists, backed initially by the major space powers, which viewed any boundary demarcation as possibly restricting their access to space, whether for peaceful or non-military purposes, considered it insufficient or inadequate to delimit a boundary of outer space without obvious scientific and technological evidences. Last more than 50 years there were large development in the exploration and use of outer space. But a large number states including those taking the view of a functionalist have taken on a negative attitude. As the element of location is a decisive factor for the choice of the legal regime to be applied, a purely functional approach to the regulation of activities in the space above the Earth does not offer a solution. It seems therefore to welcome the arrival of clear evidence of a growing recognition of and national practices concerning a spatial approach to the problem is gaining support both by a large number of States as well as by publicists. The search for a solution to the problem of demarcating the two different legal regimes governing the space above Earth has undoubtedly been facilitated and a number of countries including Russia have already advocated the acceptance of the lowest perigee boundary of outer space at a height of 100km. As a matter of fact the lowest perigee where space objects are still able to continue in their orbiting around the earth has already been imposed as a natural criterion for the delimitation of outer space. This delimitation of outer space has also been evidenced by the constant practice of a large number of States and their tacit consent to space activities accomplished so far at this distance and beyond it. Of course there are still numerous opposing views on the delineation of a outer space boundary by space powers like U.S.A., England, France and so on. Therefore, first of all to solve the legal issues faced by the international community in outer space activities like delimitation problem, there needs a positive and peaceful will of international cooperation. From this viewpoint, President John F. Kennedy once described the rationale behind the outer space activities in his famous "Moon speech" given at Rice University in 1962. He called upon Americans and all mankind to strive for peaceful cooperation and coexistence in our future outer space activities. And Kennedy explained, "There is no strife, ${\ldots}$ nor any international conflict in outer space as yet. But its hazards are hostile to us all: Its conquest deserves the best of all mankind, and its opportunity for peaceful cooperation may never come again." This speech seems to even present us in the contemporary era with ample suggestions for further peaceful cooperation in outer space activities including the delimitation of outer space.
Cryptococcus neoformans causes life-threatening meningoencephalitis in humans, but the treatment of cryptococcosis remains challenging. To develop novel therapeutic targets and approaches, signaling cascades controlling pathogenicity of C. neoformans have been extensively studied but the underlying biological regulatory circuits remain elusive, particularly due to the presence of an evolutionarily divergent set of transcription factors (TFs) in this basidiomycetous fungus. In this study, we constructed a high-quality of 322 signature-tagged gene deletion strains for 155 putative TF genes, which were previously predicted using the DNA-binding domain TF database (http://www.transcriptionfactor.org/). We tested in vivo and in vitro phenotypic traits under 32 distinct growth conditions using 322 TF gene deletion strains. At least one phenotypic trait was exhibited by 145 out of 155 TF mutants (93%) and approximately 85% of the TFs (132/155) have been functionally characterized for the first time in this study. Through high-coverage phenome analysis, we discovered myriad novel TFs that play critical roles in growth, differentiation, virulence-factor (melanin, capsule, and urease) formation, stress responses, antifungal drug resistance, and virulence. Large-scale virulence and infectivity assays in insect (Galleria mellonella) and mouse host models identified 34 novel TFs that are critical for pathogenicity. The genotypic and phenotypic data for each TF are available in the C. neoformans TF phenome database (http://tf.cryptococcus.org). In conclusion, our phenome-based functional analysis of the C. neoformans TF mutant library provides key insights into transcriptional networks of basidiomycetous fungi and ubiquitous human fungal pathogens.
Objectives : Middle aged women with depression often experience hot flush symptoms. It is still unclear about the association between depression and hot flush symptoms. Therefore, we investigated hormonal profile, functional somatic symptoms, coping styles and attitude for menopause between depressed women with hot flush and those without hot flush. Methods : This study included 33 depressed patients with hot flush symptoms and 33 depressed patients without hot flush symptoms. Hot flush was confirmed through prospective daily symptom ratings for a week. Subjects' coping styles were assessed by the stress coping checklist. Somatic symptoms were assessed by the Patient Health Questionnaire-15(PHQ-15). Sex hormonal levels were assayed by a radioimmunoassay. The Brief World Health Organization Quality of Life Assessment Instrument(WHOQOL-BREF) was used to evaluate quality of life. Results : In coping style, depressed women with hot flush symptoms used less 'problem-centered coping'($13.15{\pm}3.17$) and 'search for social support'($11.83{\pm}2.84$) than those without hot flush symptoms($15.17{\pm}3.1$, p=0.028 ; $14.25{\pm}3.22$, p=0.009 ; respectively). Depressed women with hot flush symptoms showed more negative attitude toward post-menopause, but its statistical significance was marginally insufficient(p=0.059). We did not find any group differences in sex hormonal levels and somatic complaints assessed by the PHQ-15. The score of social relationship domain of WHOQOL-BREF was significantly lower in subjects with hot flush symptoms($8.62{\pm}2.04$) than subjects without hot flush symptoms($9.71{\pm}1.65$ ; p=0.044). Conclusions : Among middle aged women with depression, the manifestation of hot flush symptoms was associated with coping styles and attitude for menopausal transition. Hot flush symptoms in depressed women negatively influence quality of life, so clinicians actively perform therapeutic approach in case of depressed patients with hot flush symptoms. In case of depressed patients who present hot flushes, cognitive behavior therapy or stress management might be an effective treatment option in company with antidepressants or hormonal treatment. Later, longitudinal study will be needed to evaluate risk factor, cause and effect associated with hot flush and depression.
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